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    18 December 2023, Volume 27 Issue 35 Previous Issue    Next Issue
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    Phosphorylation modification of alpha synuclein in serum of patients with Parkinson’s disease
    Qi Xue, Li Jiahui, Zhu Yuanfeng, Yu Lu, Wang Peng
    2023, 27 (35):  5577-5582.  doi: 10.12307/2024.205
    Abstract ( 313 )   PDF (1186KB) ( 47 )   Save
    BACKGROUND: The aggregates formed after translational modification of α-synuclein are the main pathological changes of Parkinson’s disease. α-Synuclein can penetrate the blood-brain barrier and be delivered from the central nervous system to the peripheral blood to all parts of the body, which becomes an important pathway for the pathological dissemination of Parkinson’s disease. Therefore, it is particularly important to study the changes of blood markers in patients with Parkinson’s disease to reveal the mechanism of Parkinson‘s disease as well as for early diagnosis.
    OBJECTIVE: To analyze the differences in the structural stability of phosphorylation modification sites and generated aggregates of α-synuclein in serum of patients with Parkinson’s disease.
    METHODS: A recombinant human α-synuclein prokaryotic expression system was constructed, and the protein was purified by affinity chromatography. The purity and specificity of α-synuclein monomer was detected by SDS-PAGE and western blot. Serum samples of 26 normal controls and 26 patients with Parkinson’s disease in the Department of Neurology, Affiliated Hospital of Beihua University were collected to complete the preparation of serum α-synuclein aggregates. The modified sites for phosphorylation modification of α-synuclein protein in normal serum and serum of patients with Parkinson’s disease were identified using SWATH-mass spectrometry, and protein aggregates at different sites were quantitatively analyzed. The protein aggregates with different phosphorylation modifications were purified by immunoaffinity chromatography, and then detected for stability in the serum of patients with Parkinson’s disease by western blot.
    RESULTS AND CONCLUSION: The results of SDS-PAGE and western blot showed that α-synuclein monomers with high purity and specificity were obtained. The results of the SWATH-mass spectrometry analysis showed that phosphorylation modifications occurred in both Parkinson’s disease patients and normal human serum, with significantly more phosphorylated sites in Parkinson’s disease patients than in normal humans. The phosphorylation sites in the serum of patients with Parkinson’s disease were Serine (Ser) 87, Ser129, L-tyrosine (Tyr)125, Tyr 133 and Tyr 136. After incubation in the serum of patients with Parkinson’s disease for α-synuclein, the phosphorylation modification at Ser129 accounted for 53.65% of the total phosphorylation, with 17.21%, 15.79%, 15.79%, 9.52%, and 1.03% for Tyr125, Tyr133, Tyr136, and Ser87, respectively. The serum levels of Ser129 phosphorylation-modified aggregates were detected by western blot to be more stable than those of Tyr125, Tyr133 and Tyr136 in patients with Parkinson’s disease. To conclude, the number of α-synuclein phosphorylation modification sites in the serum of patients with Parkinson’s disease was significantly higher than that of normal subjects. The aggregate structure generated by phosphorylation modification at Ser129 in the serum of patients with Parkinson’s disease was the most stable. This may provide a diagnostic marker for the early diagnosis of Parkinson’s disease.
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    Rhythmic auditory stimulation for non-acute dyskinesia in stroke patients
    Liu Shiwen, Liu Quanfeng, Yang Zhibo, Duan Changqiu, Zhang Baoying, Tan Yue, Ai Changshan, Jiang Huiqiang, Feng Xugang, Kong Nianhua
    2023, 27 (35):  5583-5588.  doi: 10.12307/2023.952
    Abstract ( 287 )   PDF (898KB) ( 72 )   Save
    BACKGROUND: In recent years, interactive rhythmical auditory stimulation has been used in motor rehabilitation after central nervous system injuries in foreign countries. In China, research on its clinical application has also been carried out. However, there are few reports on the application of interactive rhythmical auditory stimulation in the treatment of non-acute dyskinesia following stroke.
    OBJECTIVE: To investigate the effect of rhythmic auditory stimulation on non-acute dyskinesia in stroke patients.
    METHODS: A total of 130 patients with post-stroke hemiplegia, aged 40-80 years, from March 2017 to October 2019 in the rehabilitation departments of Shenzhen Hang Seng Hospital, Affiliated Hospital of Jilin Academy of Traditional Chinese Medicine, Jilin Province People’s Hospital, Jilin Hospital of Integrative Medicine and Jilin Longtan Hospital of Traditional Chinese Medicine were included. All the patients were randomly divided into experimental group (n=55) and control group (n=75). Routine rehabilitation training was performed in both groups: once a day, five times a week, for 8 weeks. The experimental group further received rhythmic auditory stimulation once a day, five times a week, for 8 weeks. Before and after training, Fugl-Meyer assessment score, Berg balance scale score, 10-meter walking speed, Tinetti gait, FAC walking function, error value of rhythmic auditory stimulus feedback, and modified Barthel index were measured in both groups.
    RESULTS AND CONCLUSION: After training, Fugl-Meyer assessment score, Berg balance scale score, 10-meter walking speed, Tinetti gait, FAC walking function, rhythmic auditory stimulus motor feedback error values and modified Barthel index were significantly improved in both groups compared with pre-training values (P=0.000). At the end of training, the Fugl-Meyer assessment score, Berg balance scale score, Tinetti gait, FAC walking function, rhythmic auditory stimulation motor feedback error values and modified Barthel index value were better in the experimental group than in the control group (P=0.000); however, there was no significant difference in 10-meter walking speed between the two groups (P > 0.05). To conclude, rhythmic auditory stimulation training is an effective rehabilitation method for stroke patients with dyskinesia during convalescence and sequelae periods. The role of rhythmic auditory stimulation on walking speed remains to be explored.
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    Differential protein expression analysis of hypertrophic and normal ligamentum flavum in patients with lumbar spinal stenosis with Qi deficiency and blood stasis
    Wang Mengshu, Zhang Yu, Zheng Zhouhang, Chen Long, You Dongchun, Guo Weifeng, Hu Fei, Chen Huan, Liu Xingming, Wu Ronghai, Zhang Yin
    2023, 27 (35):  5589-5595.  doi: 10.12307/2023.857
    Abstract ( 151 )   PDF (1668KB) ( 29 )   Save
    BACKGROUND: From the pathological mechanism of Western medicine, ligamentum flavum hypertrophy is the key pathogenic factor of lumbar spinal stenosis, and there is a lack of biological information on lumbar spinal stenosis of the Qi deficiency and blood stasis type.
    Objective: To analyze and compare differential protein expression between hypertrophic and normal ligamentum flavum in patients with lumbar spinal stenosis of the Qi deficiency and blood stasis type. 
    METHODS: Ligamentum flavum tissue samples were collected from six lumbar spinal stenosis patients with Qi deficiency and blood stasis, including three cases of ligamentum flavum hypertrophy (experimental group) and three cases of normal ligamentum flavum (control group). 4D Label free quantitative proteomic detection was performed to screen differentially expressed proteins. Gene oncology and Kyoto Encyclopedia of Genes and Genomes were used for enrichment analysis. 
    RESULTS AND CONCLUSION: There were 183 differentially expressed proteins between the two groups, including 87 up-regulated and 96 down-regulated. Gene oncology enrichment analysis showed that biological processes mainly focused on cell processes, biological regulation and response to stimuli. Cell composition was concentrated in cell, intracellular, and protein-complex species. The main molecular functions included linkage, catalytic activity and molecular function regulator. The up-regulated proteins were mainly enriched to lysosomal signaling pathway, rheumatoid arthritis signaling pathway, and Staphylococcus aureus infection signaling pathway, while the down-regulated proteins were enriched to eight signaling pathways, namely p53 signaling pathway, renal cell carcinoma signaling pathway, transforming growth factor β signaling pathway, ubiquitin-mediated protein hydrolysis signaling pathway, Ca signaling pathway, cGMP-PKG signaling pathway, hypoxia-inducible factor 1 signaling pathway, and proteoglycan signaling pathway in cancer. INHBA, MMP14, TNC, HTRA1, FGF2 were the important differentially expressed proteins in the experimental group. To conclude, there are differential protein expressions between hypertrophic and normal ligamentum flavum in patients with Qi-stagnation and blood-stasis type lumbar spinal stenosis. INHBA may be the determinant of this disease, and its mechanism may be the activation of transforming growth factor β/Smad related signaling pathway, causing ligamentum flavum hypertrophy and subsequently leading to lumbar spinal stenosis.
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    Immunoregulatory mechanism of choline acetyltransferase/alpha 7 nicotinic acetylcholine receptor/nuclear factor-kappa B signaling pathway in the pathogenesis of rheumatoid arthritis
    Li Yutong, Liu Jingshu, Li Zhen
    2023, 27 (35):  5596-5602.  doi: 10.12307/2023.889
    Abstract ( 301 )   PDF (1853KB) ( 59 )   Save
    BACKGROUND: The cholinergic anti-inflammatory pathway is widely involved in the development of rheumatoid arthritis. However, the immunoregulatory mechanism of choline acetyltransferase (ChAT)/α7 nicotinic acetylcholine receptor (α7nAChR)/nuclear factor (NF)-κB signaling pathway in the pathogenesis of rheumatoid arthritis has not been reported.
    OBJECTIVE: To investigate the immunoregulatory mechanism of ChAT/α7nAChR/NF-κB signaling pathway in the pathogenesis of rheumatoid arthritis. 
    METHODS: Thirty-two female Wistar rats were randomly divided into healthy control group, arthritis model group, vagotomy group and sham operation group, with eight rats in each group. Except for the healthy control group, other groups were treated with bovine type-all collagen and Freund’s complete/incomplete adjuvant to construct the rat arthritis model. After successful molding, the vagus nerve was severed from the left vagus nerve in the neck in the vagotomy group, while only the vagus nerve was isolated in the sham operation group. At 5 weeks after surgery, the body mass, arthritis score, joint swelling degree, pathological changes of the spleen and joint were detected. The levels of interleukin-1, interleukin-6 and tumor necrosis factor-α in serum were detected by ELISA. The mRNA and protein expression levels of ChAT/α7nAChR/NF-κB pathway core genes were detected by qRT-PCR and western blot, respectively, and analyzed by immunohistochemistry. 
    RESULTS AND CONCLUSION: Compared with the healthy control group, the body mass of rats in the arthritis model group was significantly decreased (P < 0.05), the arthritis score was significantly increased (P < 0.01), the ankle swelling degree was aggravated, the joint space was reduced with inflammatory cell infiltration, and the white pulp and germinal center of the spleen were enlarged and increased. Moreover, the levels of interleukin-1, interleukin-6 and tumor necrosis factor-α were significantly increased, and the mRNA and protein expressions of α7nAChR, ChAT, IκBα, and NF-κBp50/p65 on the joint surface were also significantly increased in the arthritis model group compared with the healthy control group (P < 0.05, P < 0.01). Compared with the arthritis model group and the sham operation group, the body mass of rats in the vagotomy group was significantly decreased (P < 0.05), the arthritis score was significantly increased (P < 0.05, P < 0.01), the swelling degree of the ankle joint was aggravated with deformity, the joint space disappeared with inflammatory cell infiltration, the white pulp and germinal center of the spleen were enlarged and merged. Compared with the arthritis model group and the sham operation group, the levels of interleukin-1, interleukin-6 and tumor necrosis factor-α were significantly increased in the vagotomy group (P < 0.05, P < 0.01), as well as the mRNA and protein expressions of α7nAChR, ChAT, IκBα, and NF-κBp50/p65 were significantly increased on the joint surface (P < 0.05, P < 0.01). To conclude, the vagus nerve regulates the cholinergic anti-inflammatory pathway and stimulates the ChAT/α7nAChR/NF-κB signaling pathway to participate in the disease progression of rheumatoid arthritis. It indicates that the cholinergic anti-inflammatory pathway may be a potential target for the treatment of rheumatoid arthritis.
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    Mechanism by which high-intensity intermittent exercise improves skeletal muscle injury and enhances exercise capacity in rats
    Zhang Liumei, Liu Jingjing, Lin Xiaoye, Liu Lin, Lu Jiao
    2023, 27 (35):  5603-5609.  doi: 10.12307/2023.890
    Abstract ( 369 )   PDF (1507KB) ( 49 )   Save
    BACKGROUND: The potential mechanism of high-intensity intermittent exercise to enhance body adaptability and improve exercise capacity may be related to the phosphorylation level of AMP-activated protein kinase (AMPK) in skeletal muscle.
    OBJECTIVE: To clarify the role of high-intensity intermittent exercise in reducing skeletal muscle damage induced by exhaustion and improving exercise capacity and to elucidate the mechanism of AMPK-mediated changes in glucose transporter protein 4 during this process.
    METHODS: Forty-five Sprague-Dawley rats were randomly divided into control group, exhaustive exercise group (EE group) and high-intensity intermittent exercise group (HIIT+EE group), with 15 rats in each group. No intervention was given in the control group. The EE group was exhausted on the treadmill at the speed of 25-28 m/min. The HIIT+EE group underwent high-intensity intermittent exercise: running on the treadmill, 4 times a day at the speed of 28 m/min, once for 10 minutes, with an interval of 10 minutes, for 3 consecutive days, and then experienced exhaustive exercise at 24 hours after the completion of high-intensity intermittent exercise to reproduce animal models of exhaustive exercise. The exercise distance of the rats was recorded. After exhaustive exercise, plasma creatine kinase and superoxide dismutase levels were measured in each group. Apoptosis of cells in gastrocnemius muscle was detected by TUNEL staining and western blot. AMPK phosphorylation in gastrocnemius muscle was detected by western blot. Glucose transporter protein 4 expression and translocation in gastrocnemius muscle was detected by immunofluorescence staining and western blot. 
    RESULTS AND CONCLUSION: The exercise distance of rats in the HIIT+EE group was greater than that in the EE group (P < 0.05). The levels of creatine kinase and superoxide dismutase were increased in the EE group compared with the control group (P < 0.05). The levels of creatine kinase and superoxide dismutase were decreased in the HIIT+EE group compared with the EE group (P < 0.05). TUNEL staining and western blot results showed that the apoptotic rate of gastrocnemius cells in the EE group was higher than that in the control group (P < 0.05), while the apoptotic rate of gastrocnemius cells in the HIIT+EE group was less than that in the EE group (P < 0.05). There was no significant difference in the expression of apoptosis-related proteins Bax and Bcl-2 between the three groups (P > 0.05). Western blot results showed that the expression of AMPK and p-AMPK protein was increased in the EE group compared with the control group (P < 0.05), while the expression of AMPK protein was decreased and the expression of p-AMPK protein was increased in the HIIT+EE group compared with the EE group (P < 0.05). Results from immunofluorescence staining and western blot assay indicated that the expression of cytosolic glucose transporter protein 4 was increased in skeletal muscle fibers in the EE and HIIT+EE groups compared with the control group. To conclude, high-intensity intermittent exercise attenuates muscle fiber damage induced by exhaustive exercise, promotes the expression and translocation of glucose transporter protein 4 in skeletal muscle by increasing the level and efficiency of AMPK phosphorylation during exercise, and thereby improves the exercise capacity of rats.
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    Differentially expressed galectin-1 during intervertebral disc degeneration
    Liang Weidong, Zhang Shuwen, Cai Xiaoyu, Xun Chuanhui, Sheng Jun, Cao Rui, Hao Honggang, Sheng Weibin
    2023, 27 (35):  5610-5615.  doi: 10.12307/2023.954
    Abstract ( 294 )   PDF (1390KB) ( 27 )   Save
    BACKGROUND: Galectin-1 is involved in the physiological and pathological processes of many tissues and organs, but its relationship with intervertebral disc degeneration remains unclear. 
    OBJECTIVE: To analyze the differential expression of galectin-1 in intervertebral discs and to investigate the effect of galectin-1 on intervertebral disc degeneration in rats.
    METHODS: Intervertebral disc specimens were collected from 15 patients in the First Affiliated Hospital of Xinjiang Medical University. Pfirrmann grading was used to definite the degree of degeneration, and the differential expression of galectin-1 in intervertebral discs with different degrees of degeneration was detected. A lumbar degeneration model was established in Sprague-Dawley rats. The model rats were randomly divided into control group, model group and galectin-1 group. Interventions were carried out on alternate days after modeling. Control and model groups were given normal saline injection into the tail vein, while galectin-1 recombinant protein was injected into the tail vein of the galectin-1 group once a day for 7 days. Eight weeks after modeling, pathological changes in the intervertebral disc were observed after Pfirrmann grading using Bruker Pharmascan 7.0T MRI and the expression of galectin-1, type II collagen and Aggrecan in the disc was examined.
    RESULTS AND CONCLUSION: qRT-PCR, immunohistochemical staining and western blot analysis indicated that galectin-1 expression decreased gradually with the aggravation of intervertebral disc degeneration, and there were significant differences among three groups (P < 0.05). At 8 weeks after modeling, MRI scan of the rat lumbar vertebra results indicated that galectin-1 could reduce the modified Pfirrmann grade of intervertebral disc degeneration, and hematoxylin-eosin staining suggested that galectin-1 could reduce the pathological manifestations of intervertebral disc degeneration. Immunohistochemistry and western blot results suggested that intervention with galectin-1 could increase the expression of galectin-1 in the intervertebral discs while reducing the degradation of type II collagen and Aggrecan, and there were significant differences among groups (P < 0.05). To conclude, the expression of galectin-1 gradually decreases with the aggravation of intervertebral disc degeneration, and the intervention with galectin-1 delays the progression of intervertebral disc degeneration in rats. 
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    Establishing a rat model of intervertebral disc degeneration by castration of both upper limbs combined with intervertebral disc puncture
    Sun Xiaoxian, Bai Xue, Liu Mengmin, Guo Yang, Lin Shun, Wu Wenxuan, Ma Yong, Liu Jintao
    2023, 27 (35):  5616-5621.  doi: 10.12307/2023.845
    Abstract ( 259 )   PDF (2224KB) ( 105 )   Save
    BACKGROUND: Seeking a relatively suitable animal model of intervertebral disc degeneration is of great significance for an in-depth study of the pathogenesis of intervertebral disc degeneration.
    OBJECTIVE: To seek a relatively suitable animal model of intervertebral disc degeneration, providing a new modeling option for basic research into degenerative disc disease.
    METHODS: Twenty male Sprague-Dawley rats were randomly divided into sham operation group and model group. The intervertebral disc degeneration model was prepared by castration of both upper limbs combined with intervertebral disc puncturing. MRI examination was performed on the day of modeling and at 4 and 8 weeks after modeling to observe the morphological changes of the intervertebral discs and evaluate the Pfirrmann grade before and after modeling. The L4-L5 and L5-L6 intervertebral discs were removed from the rats at 4 and 8 weeks after modeling. The pathological changes of the intervertebral discs were observed by hematoxylin-eosin and Masson staining and the expression of type II collagen was observed by immunohistochemistry.
    RESULTS AND CONCLUSION: The model group showed only slight degeneration on the day of modeling. Compared with the sham operation group, the T2-weighted signal on MRI was decreased in the model group, and there was a significant difference in the Pfirrmann grading at 4 weeks after modeling (P < 0.05). Compared with the sham operation group, the T2-weighted signal on MRI was significantly decreased and the boundary with the annulus fibrosa was blurred in the model group at 8 weeks after modeling. The nucleus pulposus in the model group was expanded in the posterior longitudinal ligament and there was a significant difference in the Pfirrmann grading between the two groups (P < 0.01). The results of pathological staining showed that compared with the sham operation group, the model group had a reduction in the number of nucleus pulposus cells, disordered annulus fibrosus, and decreased expression of type II collagen at the 4th week after modeling. While at the 8th week after modeling, the nucleus pulposus cells were shrunk, the matrix was condensed, the annulus fibrosus was ruptured, the structure was damaged and the boundary between the nucleus pulposus and the nucleus pulposus was blurred, and the expression of type II collagen in the nucleus pulposus was decreased more obviously in the model group compared with the sham operation group. To conclude, this method can successfully establish the rat model of intervertebral disc degeneration, and the animal model develops obvious degeneration, which provides a new idea for the study of intervertebral disc degeneration model.
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    Effects of Gubi Powder on cartilage tissue-related signaling pathways in rabbits with knee osteoarthritis
    Li Dongdong, Chen Guangyou, Li Xiaoming, Wu Xuelian, Zhu Kai, Lei Yang, Yi Lu
    2023, 27 (35):  5622-5627.  doi: 10.12307/2023.812
    Abstract ( 298 )   PDF (1342KB) ( 29 )   Save
    BACKGROUND: Activation of Wnt signaling is closely related to the pathogenesis of knee osteoarthritis. Current research on the Wnt signaling pathway has focused on the Wnt/β-catenin signaling pathway and the expression of its activation or inhibition proteins.
    OBJECTIVE: To observe the effect of Gubi Powder, an in-hospital prescription, on inflammatory factors and Notch3/Hes1 and Wnt/β-catenin signaling pathways in rabbits with knee osteoarthritis model, providing new drug ideas for clinical treatment.
    METHODS: Forty New Zealand rabbits were divided into sham operation group, model group, positive group (diclofenac potassium gel) and Gubi Powder group, with 10 rabbits in each group. The modified Hulth method was used to construct the rabbit knee osteoarthritis model, which lasted for 8 weeks. After modeling, normal saline was applied to the knee joints of the sham operation group and model group, diclofenac potassium gel was applied to the positive group, and Gubi Powder was applied to the Gubi Powder group, 8 hours a day, for 1 week. After administration, animal activity and behaviors were evaluated and scored; serum levels of prostaglandin E2, cyclooxygenase-2, interleukin 1β and tumor necrosis factor α were determined by ELISA; pathological changes of cartilage tissue were observed by hematoxylin-eosin staining and Safranin-O staining; protein levels of matrix metalloproteinases 1, 2, 3, and 13 in cartilage tissue of the knee joint were determined by immunohistochemistry; and protein levels of Notch3, Hes1, Wnt5a, β-catenin, Bcl-2, Bax, and Caspase-3 in cartilage tissue of the knee joint were determined by western blot assay. 
    RESULTS AND CONCLUSION: Compared with the sham operation group, animals in the model group had difficulties in walking, serious cartilage injury in the knee joint, significantly elevated levels of serum prostaglandin E2, cyclooxygenase 2, interleukin 1β and tumor necrosis factor α (P < 0.05), significantly increased protein expression levels of matrix metalloproteinases 1, 2, 3, and 13, Bax, Caspase-3, Wnt5a and β-catenin in cartilage tissue (P < 0.05), and decreased protein expression levels of Notch3, Hes1, and Bcl-2 (P < 0.05). Compared with the model group, the walking condition of animals in the positive group and Gubi Powder group was improved, the serum levels of prostaglandin E2, cyclooxygenase-2, interleukin 1β and tumor necrosis factor α were decreased, cartilage damage of the knee was alleviated, the protein expression levels of matrix metalloproteinases 1, 2, 3, and 13, Bax, Caspase-3, Wnt5a and β-catenin in cartilage tissue were reduced (P < 0.05), and the protein expression levels of Notch3, Hes1, and Bcl-2 were increased (P < 0.05). To conclude, Gubi Powder can alleviate the symptoms of knee osteoarthritis in rabbits, improve the pathological changes of the cartilage, and reduce inflammatory factor levels in serum. The mechanism of action may be related to the inhibition of Notch3/Hes1 and Wnt/β-catenin signaling pathways.
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    Effects of skeletal muscle massage on skeletal muscle function and conversion of skeletal muscle fiber types in type 2 diabetic rats
    Meng Meng, Hu Guanyu, Wu Xingquan, Cong Deyu
    2023, 27 (35):  5628-5633.  doi: 10.12307/2023.891
    Abstract ( 334 )   PDF (1127KB) ( 87 )   Save
    BACKGROUND: Previous studies have revealed that massage can regulate the balance of blood glucose, delay the loss of skeletal muscle mass, and promote the conversion of muscle fiber types. Skeletal muscle is also an important organ to maintain blood glucose balance and stability, but its specific association and mechanism are still unclear.
    OBJECTIVE: To investigate the effects of massage on the changes of skeletal muscle structure and function and the conversion of skeletal muscle fiber types in type 2 diabetic rats.
    METHODS: Animal models of type II diabetics mellitus were established in 24 Wistar rats by high-fat feeding combined with low-dose streptozotocin intraperitoneal injection. Eighteen rat models were randomly divided into three groups (n=6 per group): model group, metformin group and massage group. In the model group, no treatment was given; in the metformin group, metformin hydrochloride was given by gavage, once a day, 6 times per session, with an 1-day interval between sessions, for 8 sessions in total; in the massage group, massage treatment was given by small animal massager at Fenglong, Zusanli, Sanyinjiao and Blood Sea points, once a day, 6 times per session, with an 1-day interval between sessions, for 8 sessions in total. Another six healthy rats were set as blank group and received no treatment. Blood glucose and serum insulin levels, grip strength, swimming exhaustion time and skeletal muscle wet mass were measured after treatment. Western blot and qPCR were used to detect the protein and mRNA expression of myogenic determinant and myostatin in gastrocnemius muscle. ATPase staining was used to observe the changes of skeletal muscle fiber types.
    RESULTS AND CONCLUSION: Compared with the model group, fasting blood glucose and serum insulin levels after treatment were significantly reduced in the metformin group and massage group (P < 0.01). Compared with the model group, grip strength, swimming exhaustion time and skeletal muscle wet mass were increased in the massage group (P < 0.01), but there were no significant changes in the metformin group (P > 0.05). Compared with the blank group, there was a significant increase in type II muscle fibers in the model group. Compared with the model group, the number of type I muscle fibers was increased in the massage group. Compared with the blank group, the protein and mRNA expression of myogenic determinant in the gastrocnemius muscle was decreased 
    (P < 0.01), while that of myostatin in the gastrocnemius muscle was increased in the model group (P < 0.01). Compared with the model group, the protein and mRNA expression of myogenic determinant in the gastrocnemius muscle was increased (P < 0.01), while that of myostatin in the gastrocnemius muscle was decreased in the model group (P < 0.01). To conclude, massage can reduce blood glucose and improve skeletal muscle dysfunction in type 2 diabetic rats. The mechanism may be through promoting the conversion of skeletal muscle type II fibers to type I fibers, promoting the expression of myogenic determinant and inhibiting the expression of myostatin in skeletal muscle, thereby promoting the myoblastic differentiation of muscle satellite cells.
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    Proteomic study of Jintiange capsule in the treatment of retinoic acid-induced osteoporosis rats
    Zhang Chi, Zhang Xiaoyun, Chai Yuan, Chen Feng
    2023, 27 (35):  5634-5641.  doi: 10.12307/2023.823
    Abstract ( 227 )   PDF (4025KB) ( 59 )   Save
    BACKGROUND: The therapeutic effect of Jintiange capsule on primary osteoporosis has been confirmed by clinical research evidence and recommended by clinical application guidelines, but its effects and targets against secondary osteoporosis remain unclear.
    OBJECTIVE: To explore the therapeutic target and mechanism of Jintiange capsule in the treatment of secondary osteoporosis.
    METHODS: Twenty-four female Sprague-Dawley rats were randomly divided into four groups (n=6 per group): blank group (with no treatment), model group, Jintiange capsule group, and positive control group. Animal models of osteoporosis were prepared in the latter three groups by intragastric administration of retinoic acid. Fourteen days after modeling, the model group was intragastrically given normal saline, while the Jintiange capsule and positive control groups were given Jintiange capsule and alendronate sodium, respectively, for 14 days. After treatment, the femurs were taken for micro-CT scanning and tandem mass tags quantitative proteomics. Bioinformatics was used to analyze the function, signal pathway and protein interaction of the differential proteins and to screen the core proteins.
    RESULTS AND CONCLUSION: Compared with the blank group, bone volume fraction, trabecular thickness and trabecular number of the femur were significantly decreased (P < 0.05) and trabecular separation was significantly increased (P < 0.05) in the model group. Compared with the model group, bone volume fraction, trabecular thickness and trabecular number of the femur were increased (P < 0.05) and trabecular separation was decreased (P < 0.05) in the positive control and Jintiange capsule groups. A total of 2 749 differential proteins were screened between the model group and the blank group, involved in multiple biological processes such as ATP metabolism, hypoxia response, and cellular iron homeostasis as well as signal pathways such as ferroptosis, hypoxia-inducible factor 1 signal pathway, and fatty acid metabolism. A total of 773 differential proteins were screened between Jintiange capsule group and model group, involved in biological processes such as ossification, hydrogen peroxide biosynthesis, and hypoxia reaction as well as signal pathways such as hypoxia-inducible factor 1 signal pathway, fatty acid metabolism, and oxidative phosphorylation. A total of 453 targets of Jintiange capsule for treating osteoporosis were identified, including 49 primary core proteins, such as transferrin, ribosomal protein S14, and ribosomal protein S21, and 15 secondary core proteins, such as ribosomal protein S6, ribosomal protein S20, and ribosomal protein S2. These findings indicate that Jintiange capsule may negatively regulate ferroptosis by down-regulating long chain lipoacyl-CoA synthase 1, arachidonic acid-15- lipooxygenase, transferrin, and CREB binding protein to inhibit lipid peroxidation and iron accumulation, thereby modifying osteopenia and trabecular bone microstructure damage in osteoporotic rats.
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    Shikonin inhibits fibrosis of human hypertrophic scar fibroblasts via MicroRNA-382-5p
    Tang Yuting, He Xi, Wan Yu, Wang Jianjun, Yang Anning, Wu Kai, Jiao Yun, Bai Zhigang, Jiang Yideng, Shen Jiangyong
    2023, 27 (35):  5642-5648.  doi: 10.12307/2023.818
    Abstract ( 314 )   PDF (1502KB) ( 27 )   Save
    BACKGROUND: Previous studies have shown that shikonin has the potential to treat hypertrophic scar and that microRNAs are involved in the regulation of the pathological mechanism of hypertrophic scar. It is speculated that shikonin may regulate the occurrence and development of hypertrophic scar through microRNAs regulation. 
    OBJECTIVE: To investigate the mechanism of shikonin on the fibrosis of human hypertrophic scar fibroblasts via MicroRNA-382-5p. 
    METHODS: Hypertrophic scar tissue and normal skin tissue adjacent to the scar (within 3 cm around the scar) were provided by the Department of Burn and Plastic Surgery, General Hospital of Ningxia Medical University to extract human hypertrophic scar fibroblasts and human normal skin fibroblasts, respectively. Hematoxylin-eosin staining was used to identify normal skin and hypertrophic scar and immunofluorescence was used to identify fibroblasts. The relative expression level of MicroRNA-382-5p was detected by quantitative real-time PCR at the tissue level. Hypertrophic scar fibroblasts were randomly divided into shikonin group (shikonin was dissolved in dimethyl sulfone to make drug solution at a concentration of 13.46 μmol/L, which was used for cell culture for 24 hours), dimethyl sulfone group, MicroRNA-382-5p negative control group, MicroRNA-382-5p inhibitor group, shikonin+MicroRNA-382-5p negative control group and shikonin+MicroRNA-382-5p overexpression group. Real-time fluorescence quantitative PCR and western blot were used to detect the expression of Collagen Type I α1, Collagen Type III α1 and α-smooth muscle actin at mRNA and protein levels, respectively. Cell counting kit-8 was used to detect cell viability. Cell scratch test was used to detect the migration ability of cells.
    RESULTS AND CONCLUSION: Compared with normal skin fibroblasts, hypertrophic scar fibroblasts had stronger proliferative activity (P < 0.01). Compared with the dimethyl sulfone group, shikonin down-regulated the expression levels of Collagen Type I α1, Collagen Type III α1 and α-smooth muscle actin in human hypertrophic scar fibroblasts (mRNA: P < 0.01, protein: P < 0.01), and inhibited the migration of hypertrophic scar fibroblasts (P < 0.05). Compared with normal skin, MicroRNA-382-5p was highly expressed in hypertrophic scar (P < 0.01), and shikonin could down-regulate the expression of MicroRNA-382-5p (P < 0.01). Inhibition of MicroRNA-382-5p down-regulated the expression level of Collagen Type I α1, Collagen Type III α1 and α-smooth muscle actin in hypertrophic scar fibroblasts (mRNA: P < 0.01, protein: P < 0.01), and inhibited the migration of hypertrophic scar fibroblasts (P < 0.05). Under the influence of shikonin, overexpression of MicroRNA-382-5p upregulated the expression levels of Collagen Type I α1, Collagen Type III α1 and α-smooth muscle actin in hypertrophic scar fibroblasts (mRNA: P < 0.01, protein: P < 0.01), and promoted the migration of hypertrophic scar fibroblasts (P < 0.01). To conclude, shikonin can inhibit the fibrosis and migration of hypertrophic scar fibroblasts by down-regulating the expression of MicroRNA-382-5p.
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    Numerical simulation of the relationship between apparent elastic modulus and tissue elastic modulus of the trabecular meshwork in rats
    Wang Chuan, Wang Jinhang, Liu Rundong, Zhang Jing, Li Lin, Liu Zhicheng
    2023, 27 (35):  5649-5652.  doi: 10.12307/2023.856
    Abstract ( 302 )   PDF (1438KB) ( 128 )   Save
    BACKGROUND: The tissue mechanical properties of trabecular meshwork are closely related to the resistance of the aqueous humor outflow. Considering the porous structure of trabecular meshwork, the apparent elastic modulus of trabecular meshwork tissue can be obtained by atomic force microscope indentation technique. However, the information of tissue elastic modulus cannot be obtained by this method. Therefore, there is a lack of quantitative research on the relationship between apparent and tissue elastic modulus of the trabecular meshwork.
    OBJECTIVE: To further analyze the relationship between apparent and tissue elastic modulus of the trabecular meshwork by finite element simulation.
    METHODS: Based on the two-photon confocal images of rat trabecular meshwork, a three-dimensional structure model of the trabecular meshwork was constructed. The finite element analysis was used to simulate the indentation experiment to obtain the apparent elastic modulus of the trabecular meshwork at different positions, and the tissue elastic modulus was obtained by simulating the whole indentation experiment.
    RESULTS AND CONCLUSION: The parameters of the first-order Ogden model based on the atomic force microscope indentation experiment were obtained. The average value of the apparent elastic modulus of the trabecular meshwork obtained by simulating the atomic force microscope indentation experiment was within the range of the tissue elastic modulus of the trabecular meshwork tissue when the compression ratio was 1%-2%. This study quantifies the relationship between apparent and tissue elastic modulus of trabecular meshwork tissue and provides a methodological reference for the study of mechanical properties of trabecular meshwork tissue.
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    Bioinformatics analysis and experimental validation of genes related to the pathogenesis of Alzheimer's disease 
    Li Xinru, Chai Shifan, Li Weiran, Cai Hongyan, Ye Yucai, Li Shuo, Hou Meng, Wang Zhaojun
    2023, 27 (35):  5653-5658.  doi: 10.12307/2023.855
    Abstract ( 382 )   PDF (2804KB) ( 46 )   Save
    BACKGROUND: The mechanism mining of Alzheimer’s disease is very important and bioinformatics is an effective prediction technology for the potential targets of Alzheimer’s disease. 
    OBJECTIVE: To screen the genes related to the pathogenesis of Alzheimer’s disease and verify them at the animal level by bioinformatics analysis. 
    METHODS: Differentially expressed genes were screened through the GEO online database. GO/KEGG enrichment analysis was performed using the DAVID online database. A protein-protein interaction network was constructed using the STRING database. Target protein distribution in human was identified using the HPA database. Finally, protein expression was analyzed using immunofluorescence and western blot techniques. 
    RESULTS AND CONCLUSION: Gene chips for Alzheimer’s disease and healthy populations were obtained using GSE48350, a dataset within the GEO online database, and the GEO2R online analysis platform was used to analyze the differentially expressed genes between the two populations, including 42 up-regulated genes and 131 down-regulated genes. Results of the GO enrichment analysis suggested that the differentially expressed genes were mainly located in presynaptic, transport vesicles and extracellular vesicles, to mediate biological processes such as neurotransmitter release and synaptic vesicle function. KEGG pathway analysis results showed that differentially expressed genes were mainly enriched in synaptic function-related signaling pathways such as neuroactive ligand-receptor interactions, γ-aminobutyric acid synapses and retrograde endogenous cannabinoid signaling. Based on the protein interaction network, five up-regulated hub genes (CLU, GFAP, CD44, FOS, ANXA1) and five down-regulated hub genes (GABRG2, SYT1, SYN2, KCNC1, SLC32A1) were identified. It was confirmed that the expression of GABRG2, KCNC1, and SLC32A1 in the hippocampal tissue in a mouse model of Alzheimer’s disease were significantly downregulated. To conclude, these three genes can be used as potential genes for the treatment and diagnosis of Alzheimer’s disease and provide important clues for the treatment of Alzheimer’s disease. GABRG2, KCNC1 and SLC32A1 are also co-therapeutic targets for Alzheimer’s disease and epilepsy.
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    Regulatory effects of cAMP response element-binding protein on hippocampal brain-derived neurotrophic factor level in a rat model of drug-resistant epilepsy
    Li Jingxuan, Shi Dai, Wu Guofeng
    2023, 27 (35):  5659-5664.  doi: 10.12307/2023.847
    Abstract ( 280 )   PDF (992KB) ( 26 )   Save
    BACKGROUND: The formation mechanism of drug-resistant epilepsy is still unclear, and the role of cAMP response element-binding protein (CREB) and (brain-derived neurotrophic factor (BDNF) in the formation of drug-resistant epilepsy has been widely studied.
    OBJECTIVE: To regulate the expression of CREB in the hippocampus of normal rats by lentivirus transfection and to explore its effect on the formation of drug-resistant epilepsy and the expression of BDNF in the hippocampus of rats.
    METHODS: Lentivirus overexpression or interfering was used to regulate the expression level of CREB in the hippocampus of rats and detect the virus transfection results. Lithium-pilocarpine induced epilepsy models were established in rats 7 days after lentivirus injection. Sodium phenytoin and phenobarbital were then used for resistance selection. Drug-resistant epileptic rats were screened and divided into overexpression group, overexpression control group, interference group and interference control group. The frequency of epileptic seizures during the screening process was observed and the expression levels of CREB and BDNF in the hippocampus of rats were measured by western blot and real-time fluorescence quantitative PCR.
    RESULTS AND CONCLUSION: Fourteen days after virus transfection, the hippocampus of rats was covered with green fluorescence, indicating that the virus transfection level was good. The frequency of epileptic seizures in the overexpression group was higher than that in the overexpression control group and the frequency of epileptic seizures in the interference group was lower than that in the interference control group. The expression levels of CREB and BDNF in the overexpression group were higher than those in the overexpression control group, while the expression levels of CREB and BDNF in the interference group were lower than those in the interference control group. These results indicate that the regulation of CREB expression level in the hippocampus of rats can affect epileptic seizures and formation of drug-resistant epilepsy in rats, and regulating CREB expression may affect the expression level of BDNF in the hippocampus of rats with drug-resistant temporal lobe epilepsy.
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    Effect of tea polyphenols on proliferation and differentiation of osteoblasts stimulated by lipopolysaccharide
    Qin Yixiong, Yuan Zijian, Xie Shanzhou, Zhu Yingwen, Chen Minghui, Han Biao, Guo Yong
    2023, 27 (35):  5665-5669.  doi: 10.12307/2023.685
    Abstract ( 302 )   PDF (1073KB) ( 36 )   Save
    BACKGROUND: Studies have shown that tea polyphenols can play anti-inflammatory and antioxidant effects in the investigation of periodontitis and other diseases.  
    OBJECTIVE: To investigate the effects of tea polyphenols on the proliferation, differentiation and oxidative stress of mouse osteoblasts induced by lipopolysaccharide.
    METHODS: The mouse osteoblasts MC3T3-E1 were cultured in vitro, and the MC3T3-E1 cells were randomly divided into four groups, namely the control group (conventional culture), the lipopolysaccharide group (10 μg/mL lipopolysaccharide), the tea polyphenols group (1 μg/mL tea polyphenols), and the combination group (10 μg/mL lipopolysaccharide + 1 μg/mL tea polyphenols). The MTT assay was used to detect the cell proliferation. The alkaline phosphatase activity was detected by alkaline phosphatase kit. The alkaline phosphatase staining degree was detected by alkaline phosphatase azo coupling method. The calcium deposition amount of the cells was detected by calcium detection kit. The superoxide content of cells was detected by superoxide kit. The malondialdehyde content of cells was detected by malondialdehyde kit.  
    RESULTS AND CONCLUSION: (1) Compared with the control group, the proliferation activity, alkaline phosphatase activity and calcium deposition of osteoblasts were significantly decreased in the lipopolysaccharide group (P < 0.05); the staining degree of alkaline phosphatase was significantly decreased (P < 0.01); and the contents of superoxide and malondialdehyde were significantly increased (P < 0.05). The proliferation activity, alkaline phosphatase activity and calcium deposition of osteocytes in the tea polyphenols group were significantly increased (P < 0.05); the staining degree of alkaline phosphatase was significantly increased (P < 0.01); and the contents of superoxide and malondialdehyde were significantly decreased (P < 0.05). (2) Compared with the lipopolysaccharide group, the proliferation activity and calcium deposition of bone cells in the combination group were significantly increased (P < 0.05); the staining degree of alkaline phosphatase was significantly increased (P < 0.001); and the contents of superoxide and malondialdehyde were significantly decreased (P < 0.05). (3) The results showed that tea polyphenols can promote the proliferation and differentiation of osteoblasts, reduce the expression levels of oxidative stress factors, and reverse the adverse effects of lipopolysaccharides on osteoblasts.
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    Mechanism of Mongolian Medicine Erden-uril on avascular necrosis of the femoral head in rats
    Jin Hongguang, Shi Zhuang, Wang Huaxin, Ni Ma, Fu Yong
    2023, 27 (35):  5670-5675.  doi: 10.12307/2023.854
    Abstract ( 301 )   PDF (1573KB) ( 92 )   Save
    BACKGROUND: Steroid-induced avascular necrosis of the femoral head is a complication of bilateral osteonecrosis of the femoral head caused by steroid treatment with a high disability rate, but the therapeutic effect is not significant. Erden-uril is a precious Mongolian medicine prescription, mainly used to prevent and treat bone and joint diseases, but its mechanism of improving steroid-induced avascular necrosis of the femoral head is still unclear.
    OBJECTIVE: To study the action mechanism of Mongolian Medicine Erden-uril on steroid-induced avascular necrosis of the femoral head in rat.
    METHODS: Fifty rats were randomly divided into normal group, model group, low-dose Erden-uril group (0.1 g/kg Erden-uril by gavage), high-dose Erden-uril group (0.4 g/kg Erden-uril by gavage) and alendronate sodium group. Except for the normal group, rats in the other groups were intraperitoneally injected with 10 μg/kg lipopolysaccharide and 20 mg/kg methylprednisolone to establish animal models of steroid-induced avascular necrosis of the femoral head. Administration by gavage in each group was performed once a day for 6 weeks. Subsequently, hematoxylin-eosin staining was used to evaluate the femoral head necrosis model; TUNEL method was used to detect apoptosis in femoral head tissue; ELISA and immunohistochemistry were used to detect the expression levels of pro-apoptotic protein Bax and anti-apoptotic protein Bcl-2 in femoral head tissue; and western blot method was used to detect the protein expression of ATG5, Beclin-1, LC3II, Caspase-3, Bax and Bcl-2 in femoral head tissue.
    RESULTS AND CONCLUSION: Compared with the normal group, the number of empty bone lacunae was significantly increased, and adipocytes were enlarged and increased in the model group; the expression of Bax increased significantly and that of ATG5, Beclin-1, LC3II and Bcl-2 decreased significantly in the model group. Compared with the model group, treatment with Mongolian medicine Erdun-uril could significantly reduce the number of empty bone lacunae and the amount of adipocytes, significantly increase the number of bone marrow cells, and significantly decrease the number of apoptotic cells. Compared with the model group, all methods showed significant down-regulation of Bax and Caspase-3 expression and significant up-regulation of ATG5, Beclin-1, LC3II and Bcl-2 expression after treatment with low and high doses of Erdun-uril, except for western blot detection, which showed significant up-regulation of Bax expression after treatment with low-dose Erdun-uril. Moreover, there was no significant difference in protein expression between low- and high-dose Erdun-uril groups. To conclude, Erdun-uril may protect avascular necrosis of the femoral head induced by lipopolysaccharide combined with methylprednisolone in rats by promoting autophagy with anti-apoptotic effects. 
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    Enhanced miR-185-5p expression attenuates inflammatory responses in a mouse model of experimental periodontitis
    Li Hao, Ye Zhimao, Luo Yicai, Li Kongmei, Mai Yuying, Wang Qi
    2023, 27 (35):  5676-5680.  doi: 10.12307/2023.892
    Abstract ( 266 )   PDF (1256KB) ( 100 )   Save
    BACKGROUND: The pathogenesis of periodontitis is still unclear, and its therapeutic target remains to be explored. Recent studies have showed that miR-185-5p has anti-inflammatory effects in many diseases. However, it is unclear whether miR-185-5p can influence the inflammatory response in periodontitis.
    OBJECTIVE: To investigate the effect of regulating miR-185-5p expression on inflammatory response in experimental periodontitis in mice.
    METHODS: Twenty-four C57B/6J mice were randomly divided into normal control group, periodontitis control group, periodontitis miR-NC group and periodontitis miR-mimics group (n=6 per group). Except for the normal control group, other groups were induced into experimental periodontitis. Four weeks after modeling, in the periodontitis miR-mimics group, 100 μL of normal saline containing 0.5 mmol/L miR-185-5p mimics was injected into the proximal and distal gingival papillae of bilateral mandibular second molars of mice. In the periodontitis miR-NC group, 100 μL of normal saline containing 0.5 mmol/L negative control miR-NC was injected. In the normal control group and the periodontitis control group, the same amount of normal saline was injected. After 8 weeks of administration, all mice were sacrificed. Micro-CT was used to detect the left mandibles, and qRT-PCR was used to detect the gene expression of miR-185-5p, marginal zone B and B-1 cell-specific protein (MZB1), tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) in the gingiva of left mandibles. Immunohistochemical staining was used to detect the protein expression of MZB1, phospho-p65 NF-κB (pp65), TNF-α and IL-6 in the gingiva of right mandibles. 
    RESULTS AND CONCLUSION: Micro-CT examination results showed that compared with the periodontitis control group and periodontitis miR-NC group, alveolar bone loss in the periodontitis miR-mimics group was decreased (P < 0.05). qRT-PCR results showed that the expression of miR-185-5p in the periodontitis miR-mimics group was higher than that in the normal control group, periodontitis control group, and periodontitis miR-NC group (P < 0.05). Compared with the periodontitis control group and periodontitis miR-NC group, the mRNA expression of MZB1, TNF-α, and IL-6 was decreased in the periodontitis miR-mimics group (P < 0.05). Immunohistochemical staining results showed that compared with the periodontitis control group and periodontitis miR-NC group, the protein expression of MZB1, pp65, TNF-α and IL-6 was decreased in the periodontitis miR-mimics group (P < 0.05). Therefore, increasing the expression of miR-185-5p can inhibit the inflammatory response in periodontitis and the resorption of alveolar bone, which may be associated with the suppression of MZB1 and nuclear fctor-κB pathway in the gingival tissue.
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    Shutiao Jingjin Massage can stabilize intracellular environment of rabbit chondrocytes following knee osteoarthritis-induced cartilage injury
    Zheng Lijun, Wang Kai, Li Muzhen, Wang Jianmin, Qiao Yingjie, Li Huadong
    2023, 27 (35):  5681-5687.  doi: 10.12307/2023.819
    Abstract ( 269 )   PDF (1613KB) ( 32 )   Save
    BACKGROUND: It has been found that Shutiao Jingjin Massage can effectively improve the clinical symptoms of knee osteoarthritis in the early stage, but its treatment mechanism needs to be verified.
    OBJECTIVE: To explore the effect of Shutiao Jingjin Massage on knee osteoarthritis and its possible mechanism
    METHODS: Thirty-four male New Zealand rabbits were randomly divided into blank group (n=9) and model group (n=25). Rabbit models of knee osteoarthritis were established by injecting 4% papain solution into the knee joint cavity. Six weeks after modeling, one rabbit in each group was randomly selected for cartilage hematoxylin-eosin staining and Mankin’s scoring. After the model evaluation, the model group was randomly divided into model control group (n=8), massage group (n=8) and glucosamine group (n=8). The massage group was treated with Shutiao Jingjin Massage, once every other day; the glucosamine group was given aqueous solution of glucosamine sulfate capsules by gavage, once a day. Interventions in each group lasted 4 weeks. At 1 week after intervention, the pathological morphology of cartilage was observed by safranin O-fast green staining. The expression of extracellular signal-regulated kinases 1 and 2, nuclear factor-kappa B p65, Bcl-2, Bax, and cysteine-aspartic acid protease-3 in cartilage tissue was detected by western blot assay and immunohistochemistry. The levels of interleukin-1β in peripheral blood and joint fluid were determined by enzyme-linked immunosorbent assay.
    RESULTS AND CONCLUSION: Cartilage section microscopic observation: the model control group had cartilage surface defect and damage, cartilage layer thinning, abnormal cluster aggregation of chondrocytes, and tidal line damage; in the massage group, there was no obvious damage on the cartilage surface, chondrocyte arrangement was basically normal, no obvious abnormal aggregation occurred, and the tidal line was regular; compared with the model control group, the glucosamine group did not show significant improvement. Compared with the blank group, the expression of extracellular signal-regulated kinases 1 and 2, nuclear factor-kappa B p65, Bax, and cysteine-aspartic acid protease-3 protein in the model control group was increased (P < 0.01), while the Bcl-2 expression decreased (P < 0.01). The expression levels of extracellular signal-regulated kinases 1 and 2, nuclear factor-kappa B p65, Bax, and cysteine-aspartic acid protease-3 protein in the massage group were significantly lower than those in the model control group (P < 0.01) and slightly lower than those in the glucosamine group (P < 0.05), while the expression of Bcl-2 was significantly higher than that in the model control group (P < 0.01) and slightly higher than that in the glucosamine group (P < 0.05). Compared with the blank group, the mass concentrations of interleukin-1β in the joint fluid and peripheral blood were significantly increased in the model control group (P < 0.01). Compared with the model control group, the mass concentrations of interleukin-1β in the joint fluid and peripheral blood were significantly decreased in the massage group (P < 0.01). The mass concentrations of interleukin-1β in the joint fluid and peripheral blood were significantly higher in the glucosamine group than the massage group (P < 0.01). To conclude, Shutiao Jingjin Massage can inhibit cartilage degeneration and slow down the progression of knee osteoarthritis, probably by regulating the secretion and release of interleukin-1β, interacting with the extracellular signal-regulated kinase 1/2-nuclear transcription factor κB signaling pathway, inhibiting excessive apoptosis of chondrocytes, maintaining the stability of intracellular environment in chondrocytes and repairing damaged cartilage.
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    Acupoint catgut embedding regulates the role of type II alveolar epithelial cells in airway inflammation in asthmatic rats
    Tang Xuyun, Chen Panbi, Du Dijia, Qin Zhongyin, Long Runjin
    2023, 27 (35):  5688-5694.  doi: 10.12307/2023.869
    Abstract ( 265 )   PDF (1590KB) ( 25 )   Save
    BACKGROUND: Acupoint catgut embedding can alleviate airway inflammation in asthma, which is related to regulating the imbalance of Th1/Th2 in the p38 MAPK pathway. Whether the above process is affected by the functional state of type II alveolar epithelial cells at the same time remains to be further studied.
    OBJECTIVE: To observe the effect of acupoint catgut embedding on p38 MAPK pathway, interleukin-4, γ-interferon and type II alveolar epithelial cells in lung tissue of asthma rats.
    METHODS: Forty male Wistar rats were randomly divided into blank control group, asthma model group, dexamethasone group and acupoint catgut embedding group (n=10 per group). Asthma models of asthma were prepared by intraperitoneal injection of ovalbumin and aluminum hydroxide suspension on days 1 and 8 and aerosol inhalation of ovalbumin solution from day 15 was given to induce symptoms of airway inflammatory responses, once a day for 2 weeks, in the latter three groups. Meanwhile, the dexamethasone group began to receive intraperitoneal injection of dexamethasone on day 15, once a day for 2 weeks; and the acupoint catgut embedding group received embedding treatment at Feishu, Dingchuan, and Danzhong acupoints on day 15. At the end of aerosol inhalation, the relative content of eosinophils in the alveolar lavage fluid was counted by Richter-Kimsa staining, the ultrastructural changes of type II alveolar epithelial cells in lung tissue were observed by transmission electron microscopy, the positive expression of interleukin-4 and γ-interferon in lung tissue was detected by immunohistochemistry, and the protein and gene expression of p-p38 MAPK, interleukin-4 and γ-interferon in lung tissue were detected by western blot and PCR respectively.
    RESULTS AND CONCLUSION: Compared with the asthma model group, the relative content of eosinophils in the bronchoalveolar lavage fluid was significantly reduced in the dexamethasone group and acupoint catgut embedding group (P < 0.01), while compared with the dexamethasone group, the relative content of eosinophils in the bronchoalveolar lavage fluid was significantly higher in the acupoint catgut embedding group (P < 0.05). Under the transmission electron microscopy, damage to lamellar vesicles and mitochondria in type II alveolar epithelial cells was alleviated in the dexamethasone and acupoint catgut embedding groups compared with the asthma model group. Immunohistochemical detection showed that compared with the asthma model group, the expression of interleukin-4 in lung tissue was decreased (P < 0.01) and that of γ-interferon was increased (P < 0.01) in the dexamethasone and acupoint catgut embedding group; compared with the dexamethasone group, the expression of interleukin-4 in lung tissue was increased (P < 0.01) and that of γ-interferon was decreased (P < 0.01) in the acupoint catgut embedding group. Compared with the asthma model group, the protein and mRNA expressions of p-p38 MAPK and interleukin-4 in the lung tissue of the dexamethasone and acupoint catgut embedding groups were decreased (P < 0.01, P < 0.05), while the protein and mRNA expressions of γ-interferon were increased (P < 0.01). To conclude, acupoint catgut embedding may upregulate γ-Interferon and downregulate interleukin-4 by inhibiting the p38 MAPK signal pathway, thereby reducing stress damage in type ii alveolar epithelial cells and alleviating the airway inflammatory response in asthma.
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    The power-law model-based analysis of aerobic exercise against Alzheimer’s disease in animal experiments
    Tang Lu, Lin Haiqi
    2023, 27 (35):  5695-5700.  doi: 10.12307/2023.861
    Abstract ( 286 )   PDF (939KB) ( 88 )   Save
    BACKGROUND: Yin-Yang and the golden mean are the core concepts of Chinese culture, but there is a lack of a bridge connecting with modern science. The power-law model has good robustness or anti-interference ability in complex system science, suggesting that it is closely related to Yin-Yang and the golden mean.
    OBJECTIVE: To analyze the Yin-Yang dialectical characteristics of long-term aerobic exercise against Alzheimer’s disease based on the power-law model and to discuss the potential role of Yin-Yang theory of power-law model in the complex systems of the human body.
    METHODS: Referring to the theory of Yin-Yang, the relationship rationality and the golden mean, this paper uses power law to characterize each pair of parameters, and the factors that cause the power law coefficient and index changes are referred to as Yin-Yang causes. Then, a parametric pair model of human Yin-Yang is proposed and applied to experimental animal studies of Alzheimer’s disease. The parameters of the human body constitute the parameter space, and the geometric mean of the parameter space is defined as the golden center of the parameter space.
    RESULT AND CONCLUSION: Yin and Yang are subjective feelings of power-law relationship of objective parameters. Yin-Yang attribute of two subspaces exists in relation to the joint space of two subspaces. Yin-Yang secret or Yin-Yang imbalance in two subspaces correspond to the same-level power law or upgraded power law in two subspaces respectively. Animal experimental data shows that Alzheimer’s disease modeling can cause Yin-Yang imbalance, and habitual swimming before modeling can resist the changes in Yin-Yang parameters caused by Alzheimer’s disease modeling. The above results indicate that the reasonable explanation of Yin-Yang theory of power-law model by human complex system parameters can be used for Yin-Yang dialectic in data space and the power-law relationship in objective parameters can represent the Yin-Yang characteristics of subjective feelings.
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    Finite element analysis of knee joint stress in stop-jump maneuvers after anterior cruciate ligament reconstruction
    Liu Yihui, Yan Ke, Zhang Liwen, Zhang Meizhen, Wu Xiaogang, Chen Weiyi
    2023, 27 (35):  5701-5706.  doi: 10.12307/2023.853
    Abstract ( 317 )   PDF (2081KB) ( 57 )   Save
    BACKGROUND: The kinematic and kinetic characteristics of the knee joint during the different movements of patients undergoing anterior cruciate ligament reconstruction have been analyzed. Herein, it is assumed that patients undergoing anterior cruciate ligament reconstruction have higher von Mises stress in the anterior cruciate ligament on the unaffected side than on the affected side and lower von Mises stress in the meniscus and femoral cartilage on the unaffected side than on the affected side during stop-jump maneuvers.
    OBJECTIVE: To investigate the stress response characteristics of the soft tissue of the knee on the unaffected and affected sides of patients undergoing anterior cruciate ligament reconstruction in order to provide a reference for reducing secondary injuries such as secondary anterior cruciate ligament injuries and chronic knee osteoarthritis in this population.  
    METHODS: Kinematic and kinetic parameters of the knee joint were acquired during the stop-jump maneuvers in patients with anterior cruciate ligament reconstruction using an infrared spot motion capture system (Nokov) and a force measuring platform (Bertec). The three-dimensional angle and torque parameters of the knee joint were obtained by the Euler angle calculation method and inverse dynamics. The three-dimensional angle and torque, used as boundary and loading conditions, were then loaded into the finite element model of the knee joint, and numerical simulations were subsequently performed to compare the stress distribution of the internal structure of the knee joint on the unaffected and affected sides under actual motion loads.
    RESULTS AND CONCLUSION: At the peak of the first horizontal posterior ground reaction force, the flexion and adduction angles of the affected knee were significantly greater than those of the unaffected side, and the vertical ground reaction force was higher on the unaffected side than on the affected side. In addition, the peak vertical ground reaction force of the unaffected side appeared when the stop-jump maneuver was performed by 4%-6%, and that of the affected side appeared when the stop-jump maneuver was performed by 15%-17%, showing the unaffected leg lands earlier than the affected side. Finite element analysis results showed that the peak value of anterior cruciate ligament von Mises stress on the unaffected side was higher than that on the affected side (28.47 MPa vs. 13.18 MPa). The maximum stress on the unaffected side occurred at the posterior tract of the anterior cruciate ligament, while that on the affected side was mainly distributed in the anterior femoral end of the anterior cruciate ligament. In addition, the von Mises stress peak values of the affected femoral cartilage (12.16 MPa vs. 5.342 MPa) and meniscus (17.35 MPa vs. 16.18 MPa) were greater than those of the unaffected side. The corresponding maximum stress peaks were located at the edge of the lateral condyle of the distal femoral articular cartilage and the edge of the anterior horn of the lateral meniscus, respectively. Finite element simulation indicated that the risk of anterior cruciate ligament injury on the unaffected side might be greater than that on the affected side. Furthermore, knee osteoarthritis could be aggravated by a higher stress on the affected femoral cartilage and meniscus. Overall, the rehabilitation training programs for patients undergoing anterior cruciate ligament reconstruction should not only pay attention to the rehabilitation training of the affected knee, but also focus on the abnormal movement pattern and compensation mechanism of the unaffected knee.
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    Regulation of cardiovascular diseases by histone deacetylation modification
    Han Weiyu, Chen Yuanxing, Huang Youyang, Liu Weiwei, Zhao Yongchao, Zhao Ranzun
    2023, 27 (35):  5707-5713.  doi: 10.12307/2023.599
    Abstract ( 379 )   PDF (1158KB) ( 47 )   Save
    BACKGROUND: Histone modification is a reversible post-translational modification that acts as a marker of transcriptional activation or inhibition and controls cell metabolism, damage, and repair. Deacetylation, as one of the histone posttranslational modifications, plays an important role in the occurrence and development of cardiovascular diseases. 
    OBJECTIVE: To mainly review the mechanisms of histone deacetylation in atherosclerosis, myocardial hypertrophy, heart failure, myocardial infarction, and hypertension, in order to further understand the pathological processes related to the occurrence and development of cardiovascular diseases, providing theoretical reference for the diagnosis, treatment and prognosis of cardiovascular diseases. 
    METHODS: Related articles published from 1979 to 2022 were retrieved from PubMed, Web of Science and CNKI database. The keywords were “histone, histone modification, histone acetylation, histone deacetylation, histone transacetylase, histone deacetylase, cardiovascular, atherosclerosis, cardiac hypertrophy, heart failure, myocardial infarction, hypertension” in English and Chinese. Ultimately, we included 64 articles for review.
    RESULTS AND CONCLUSION: Histone acetylation modifications occur mainly at the more conserved N-terminal lysine residues of core histones and are regulated synergistically by histone acetyltransferases and histone deacetylases, which have emerged as an important form of epigenetic regulation. Histone deacetylation modifications regulate the occurrence and development of cardiovascular diseases and the following conclusions can be drawn: (1) At the protein level, histone deacetylation modifications increase the diversity and complexity of mechanisms between cellular pathways, mainly by changing the localization, activity or function of proteins, which then affect a variety of important cellular life activities, thus playing an important role in the diagnosis, treatment and prognosis of cardiovascular diseases. (2) Histone acetyltransferase and histone deacetylase molecules are closely related to cardiovascular diseases such as atherosclerosis, myocardial hypertrophy, myocardial infarction and hypertension, and their corresponding inhibitors can be used as targeted drugs for these diseases. (3) Research on histone acetylation and deacetylation modifications in cardiovascular diseases is still at the basic research stage and the specific mechanisms of histone deacetylation modifications in some diseases are still being investigated. As research progresses, new breakthroughs in histone acetylation modifications are expected in the treatment of cardiovascular disease and in clinical translation.
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    Value of a critical bone defect animal model in evaluating osteogenic efficacy of bone tissue engineering scaffold
    Xiong Wei, Yuan Lingmei, Qian Guowen, Huang Jinyang, Pan Bin, Guo Ling, Zeng Zhikui
    2023, 27 (35):  5714-5720.  doi: 10.12307/2023.852
    Abstract ( 307 )   PDF (1016KB) ( 123 )   Save
    BACKGROUND: The repair of critical size bone defects has always been a challenge clinical problem for trauma orthopedic surgeons. With the development of clinical techniques and biomaterials, Ilizarov, Masquelet and bone tissue engineering techniques are gradually replacing the traditional autologous and allogeneic bone transplantation to become the research hotspots in the field of bone repair. Among them, bone tissue engineering scaffolds have the greatest potential for development. But a lot of basic research is necessary before the formal clinical application of bone tissue engineering scaffolds, and animal experiments are the most important. Therefore, it is very important to select an appropriate animal model of critical bone defects for testing the osteogenic efficacy of bone tissue engineering scaffolds.
    OBJECTIVE: To systematically summarize the advantages and disadvantages of different types of animal models of critical bone defects and to evaluate the osteogenic efficacy of various types of bone tissue engineering scaffolds in the animal models of critical bone defects.
    METHODS: CNKI, WanFang, Web of Science, Cochrane Library, and PubMed were searched for relevant literature. Search terms included “bone defects, animal models, bone tissue engineering, scaffolds, bone repair, bone regeneration” in Chinese and English. A total of 62 articles addressing the animal models of critical bone defects and their application to evaluate the osteogenic efficacy of bone tissue engineering scaffolds were included for further review. 
    RESULTS AND CONCLUSION: The specific definition of critical bone defects is still unclear and there is no consensus on how to establish critical defects in various animal models of bone defects. By searching the published articles about the establishment of critical bone defect animal models in mice, rats, rabbits, pigs, dogs and sheep, we found that domestic and foreign scholars basically followed the principle that the size of the bone defect that could be not self-healing during the experimental period is selected when establishing the critical bone defect model, but there were differences in the specific values. In general, rat skull, femur and rabbit femoral condyles are the more suitable sites for critical bone defect models, which are suitable for large-scale modeling because of their wide sources, low requirement of experimental environment, easy breeding, low modeling cost, and high operability. However, compared with large animals such as sheep and pigs, they have the disadvantages of small bones and the composition of bone tissue is different from that of human, which makes it difficult to fully simulate the human bone healing process. After the biocompatibility and osteogenic activity of the bone tissue engineering scaffolds are evaluated by cytological experiments, its osteogenic efficiency and biological safety in vivo can be confirmed with the help of the stable animal model of critical bone defects. The critical bone defect models of rat skull and femur and rabbit femoral condyle are respectively suitable for the preliminary evaluation of intramembranous osteogenesis and endochondral osteogenesis, while the critical bone defect models of sheep and pig tibia are ideal animal models that are recommended for preclinical evaluation. Further research on the molecular mechanism of bone tissue engineering scaffold osteogenesis is expected to help develop artificial scaffold materials that can match the osteogenesis efficiency of autogenous bone, which is of great significance for overcoming the difficult clinical problem of segmental bone defects.
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    Traditional Chinese medicine regulates nuclear factor-kappa B signaling pathway against ischemia-reperfusion injury of skin flaps
    Ma Suilu, He Zhijun, Liu Tao, Li Jinpeng, Li Yan, He Bo, Wei Xiaotao, Wang Weiwei
    2023, 27 (35):  5721-5726.  doi: 10.12307/2023.828
    Abstract ( 261 )   PDF (1108KB) ( 79 )   Save
    BACKGROUND: In recent years, it has been found that nuclear factor-κB signaling pathway aggravates ischemia-reperfusion injury of skin flaps by regulating downstream cytokines, and traditional Chinese medicine can inhibit the activation of nuclear factor-κB, thereby reducing ischemia-reperfusion injury of skin flaps.
    OBJECTIVE: To review the research progress in the regulation of nuclear factor-κB signaling pathway by Chinese medicine, its active ingredients and Chinese medicine compound against skin flap ischemia-reperfusion injury.
    METHODS: The first author searched for the relevant literatures published in CNKI and PubMed databases from January 2012 to October 2022 using the keywords of “Traditional Chinese Medicine (TCM), NF-κB, Skin flaps, vascular endothelial cell (VEC)” in Chinese and English, respectively. After screening, 45 articles were finally summarized for review.
    RESULTS AND CONCLUSION: Activated nuclear factor-κB signaling pathway can promote the release of inflammatory factors and adhesion molecules in tissue cells to anti-fibrinolytic and coagulant substances through the transcription of a variety of cytokines, and aggravate the inflammatory cascade reaction between ischemia-reperfusion-damaged flap tissue and vascular endothelial cells, leukocyte adhesion and microthrombus formation, leading to flap necrosis. Traditional Chinese medicine, its compounds and effective components (flavonoids, terpenoids, saponins, phenols, etc.) inhibit the activation of nuclear factor-κB signaling pathway, alleviate the inflammatory cascade reaction between the ischemia-reperfusion-damaged flap and vascular endothelial cells, leukocyte adhesion and thrombosis, and thereby inhibit ischemia-reperfusion injury of skin flaps. Research on the mechanism of nuclear factor-κB signaling pathway regulated by traditional Chinese medicine against ischemia-reperfusion injury of skin flaps provides a new idea for the drug research and development of skin flap ischemia-reperfusion injury, and provides reference and theoretical basis for the clinical prevention and treatment of skin flap ischemia-reperfusion injury by traditional Chinese medicine and the acceleration of skin flap healing. 
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    Role and advance of mitophagy in spinal cord injury
    Ding Yiqun, Li Xigong, Pan Wenming, Zhang Qin
    2023, 27 (35):  5727-5733.  doi: 10.12307/2023.712
    Abstract ( 447 )   PDF (1009KB) ( 51 )   Save
    BACKGROUND: Spinal cord injury is a serious disease of the central nervous system, characterized by weak regeneration of neurons and complex pathological processes. Mitochondrial autophagy is a highly selective autophagy that can degrade damaged mitochondria and plays an important role in energy supply, cell metabolism and neuronal survival.  
    OBJECTIVE: To explore the mechanism of mitochondrial autophagy and the effects of drugs on mitochondrial autophagy, and provide a new target for the treatment of spinal cord injury.
    METHODS: In CNKI, Wanfang and PubMed databases, “spinal cord injury, mitophagy, autophagy, mitochondria” were used as search terms. We searched for articles on the role and mechanism of mitophagy and spinal cord injury.  
    RESULTS AND CONCLUSION: (1) The regulation of mitochondrial autophagy level plays a key role in the treatment of spinal cord injury, and its regulatory mechanism is complex, mainly including PINK1/Parkin, Nix/BNIP3L, FUNDC1, and Atg proteins. In addition, GIT1 regulatory pathway and down-regulated miRNA-124 induction pathway were also found. (2) In terms of treatment, rapamycin, acetyl-L-carnitine, salidroside, betulinic acid, and maltol can inhibit apoptosis and improve spinal cord injury by appropriately activating mitochondrial autophagy. In contrast, rosiglitazone ameliorates spinal cord injury by inhibiting mitochondrial autophagy. This suggests that activation or inhibition of mitochondrial autophagy may improve spinal cord injury, which is related to the different mechanisms of drug action. The specific mechanism of action of the drugs remains to be further studied.
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    A systemic review of the effectiveness and safety of fractional CO2 laser combined with platelet-rich plasma in the treatment of atrophic acne scars
    Song Li, Lu Mao, Tang Yi, Liu Yanlin
    2023, 27 (35):  5734-5740.  doi: 10.12307/2023.841
    Abstract ( 295 )   PDF (1112KB) ( 116 )   Save
    OBJECTIVE: Atrophic scar is the most common complication of acne, which has a great impact on patients' psychology and quality of life. Fractional CO2 laser combined with platelet-rich plasma shows promising therapeutic prospects. This study systematically evaluated the effectiveness and safety of fractional CO2 laser combined with platelet-rich plasma in the treatment of atrophic acne scars. 
    METHODS: The Cochrane Library, PubMed, Web of Science, EMbase, CNKI, WanFang, VIP and Chinese Biomedical Literature Database were searched for literature related to clinical trials on fractional CO2 laser combined with platelet-rich plasma for the treatment of atrophic acne scar up to September 2022. Manual retrieval was conducted to complete the data. The included studies were screened and evaluated for literature quality by two researchers, and finally Meta-analysis was performed using Revman 5.3 software for the included studies, and descriptive analysis was used for data that could not be Meta-analyzed.
    RESULTS: A total of 13 studies involving 642 patients were included. Meta-analysis results showed that compared with the fractional CO2 laser alone group, the fractional CO2 laser combined with platelet-rich plasma group had significantly better overall response rate (odds ratio (OR)=3.95, 95% confidence interval (CI)=2.23 to 7.00, P < 0.000 01), better patient satisfaction (OR=4.19, 95% CI=1.05 to 16.76, P=0.04), shorter duration of erythema and edema (mean difference (MD)=-0.98, 95% CI=-1.19 to -0.77, P < 0.000 01; MD=-0.92, 95% CI=-1.36 to -0.48, P < 0.000 1 respectively), less post-inflammatory hyperpigmentation (OR=0.21, 95% CI=0.06 to 0.70, P=0.01), and shorter recovery time and total downtime (MD=-2.79, 95% CI=-3.28 to-2.30, P < 0.000 01; MD=-2.68, 95% CI=
    -3.15 to -2.21, P < 0.000 01, respectively).
    CONCLUSION: Fractional CO2 laser combined with platelet-rich plasma is more effective than fractional CO2 laser alone, with faster post-operative recovery and less adverse reactions such as erythema, edema and hyperpigmentation.
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