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    18 January 2021, Volume 25 Issue 2 Previous Issue    Next Issue
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    Role and mechanism of focal adhesion kinase in inducing osteogenic differentiation of mouse embryonic fibroblasts cells
    Li Zhen, Huang Yonghui, Sun Jifu, Sun Haitao
    2021, 25 (2):  165-171.  doi: 10.3969/j.issn.2095-4344.2969
    Abstract ( 435 )   PDF (944KB) ( 38 )   Save
    BACKGROUND: Focal adhesion kinase (FAK) is regarded as a bridge molecule of “biomaterial/scaffold,” “seed cell,” and “growth factor” in bone tissue engineering. Exploration on the role and mechanism of focal adhesion kinase in inducing osteogenic differentiation of related seed cells is particularly important for the development and application of bone tissue engineering.
    OBJECTIVE: To determine the role and mechanism of FAK in inducing osteogenic differentiation of immortalized mouse embryonic fibroblasts (iMEF). 
    METHODS: Under the same induction conditions, the iMEF cells with (iMEFFAK+/+ cells) or without FAK knockout (iMEFFAK-/- cells), treated with or without PI3K/AKT phosphorylation inhibitor LY294002 or ERK1/2 phosphorylation inhibitor U0126, were induced to differentiate into osteoblasts. The morphological changes of iMEFs (iMEFFAK+/+ and iMEFFAK-/-) at different induction periods were observed under a microscope. Runx2 protein levels and corresponding p-ERK1/2 and p-AKT levels were detected by western blot. RT-PCR technology was used to detect the transcription level of Runx2 gene. Finally, the induced iMEFs (iMEFFAK+/+ and iMEFFAK-/-) were stained with alizarin red staining for calcium nodules 3 weeks after osteogenesis induction. 
    RESULTS AND CONCLUSION: The osteogenic effect of iMEFFAK-/- cells was lower than that of iMEFFAK+/+ cells under the same induction conditions. Both the expression levels of Runx2 and the osteogenic effect of iMEFFAK+/+ cells and iMEFFAK-/- cells treated with LY294002 decreased significantly compared with the control group. Both the expression levels of Runx2 and the osteogenic effect of iMEFFAK+/+ cells and iMEFFAK-/- cells treated with U0126 decreased significantly compared with the control group. To conclude, silencing FAK expression can inhibit osteogenic differentiation of mouse embryonic fibroblasts by reducing the levels of PI3K/AKT, serine/threonine protein kinase, and ERK1/2 phosphorylation levels

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    Mechanism underlying the interventional effect of Panax Notoginsenosides on ovariectomized osteoporotic fracture rats
    Hu Guang, Guan Zhiyu, Zhang Kaiwei
    2021, 25 (2):  172-177.  doi: 10.3969/j.issn.2095-4344.2970
    Abstract ( 316 )   PDF (821KB) ( 175 )   Save
    BACKGROUND: Osteoporotic fractures are the most common serious complications caused by osteoporosis, and their repair is more difficult, which seriously threatens the health and quality of life of patients. 
    OBJECTIVE: To investigate the interventional effect of Panax Notoginsenosides in ovariectomized osteoporotic fracture rats and the relevant mechanism. 
    METHODS: Fifty healthy female rats were selected. Ten of them were normal group, and the other 40 rats were used to make ovariectomized osteoporotic fracture models. Model rats were divided into model group, low, medium and high dose group. Normal group and model group rats were intragastrically administered normal saline, rats in the low, medium and high dose group were given intragastric administration of 10, 20 and 40 mg/kg Panax Notoginsenosides solution. The study protocol was approved by the Animal Ethic Committee of the First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, No. (2019)58 on September 23, 2019.
    RESULTS AND CONCLUSION: The bone mineral density and bone cell index of the high dose group were higher than those of low and middle dose groups             (P < 0.05). The bone volume fraction, the number of callus trabeculae and the thickness of trabeculae in the high dose group were all higher than those in the model group, the low dose group and the middle dose group, and the resolution of trabeculae was lower than that in these three groups (P < 0.05). The levels of glutathione peroxidase, bone morphogenetic protein 2, and vascular endothelial growth factor in the high dose group were lower than those in the normal group, but higher than those in the model group, low and medium dose groups. The levels of lipid peroxide, malondialdehyde, osteocalcin, type I procollagen carboxy terminal propeptide, bone-specific alkaline phosphatase in the high dose group were higher than those in the normal group, but lower than those in the model group, low and medium dose group (P < 0.05). The expression of PI3K, Akt and mTOR in the low, middle and high dose groups was lower than that in the normal group and higher than that in the model group; and the expression of PI3K, Akt and mTOR in the high dose group was higher than that in the low and middle dose groups (P < 0.05). Therefore, treatment with Panax Notoginsenosides can increase the bone miner density and bone cell index, promote the growth of bone trabecula at the callus, alleviate oxidative stress injury, regulate the PI3K / Akt / mTOR signal pathway, and accelerate the formation of new blood vessels in the callus and fracture healing in ovariectomized osteoporotic fracture rats. 
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    Involvement of ERK5 signaling pathway in osteoporosis development in mice
    Geng Bin, Xia Yayi
    2021, 25 (2):  178-185.  doi: 10.3969/j.issn.2095-4344.2955
    Abstract ( 381 )   PDF (1050KB) ( 57 )   Save
    BACKGROUND: Although extracellular signal-regulated kinase 5 (ERK5) is an essential transducer of external signals for osteoblasts growth, differentiation, and survival, the effects of ERK5 signaling on bone homeostasis in vivo have not yet been described. 
    OBJECTIVE: To elucidate whether the inhibition of ERK5 activity and its downstream targets by XMD8-92 results in osteoporosis and aggravates dexamethasone-induced osteoporosis.
    METHODS: In vivo experiment: A mouse model of osteoporosis was induced by dexamethasone. There were four groups: blank control group (PBS); XMD8-92 group (50 mg/kg); dexamethasone group (50 mg/kg); XMD8-92+dexamethasone group (50 mg/kg XMD8-92 plus 50 mg/kg dexamethasone). Administration was done for 5 continuous weeks. Cell experiment: There were four groups: blank control group; XMD8-92 group (5 μmol/L); dexamethasone group (10×10-6 mol/L); XMD8-92+dexamethasone group (treatment with 5 μmol/L XMD8-92 followed by 10×10-6 mol/L dexamethasone). Cells were incubated for 1 hour. In addition, ERK5 activation was induced using epidermal growth factor. ERK5 activity in mice and MC3T3-E1 cells were detected. We further detected bone mass, bone architecture and mechanical stress of the trabecular bone, measured the expression of receptor activator of nuclear factor-κB ligand/osteoprotegerin (RANKL/OPG) and observed the roles of XMD8-92 in osteoblast proliferation and apoptosis. The study protocol was approved by the Ethics Committee of the Second Hospital of Lanzhou University (No. 2016-D44). 
    RESULTS AND CONCLUSION: XMD8-92 could block ERK5 phosphorylation, change the architecture of the trabecular bone, reduce bone mass and aggravate dexamethasone-induced osteoporosis. XMD8-92 could significantly reduce the biomechanical properties of bone tissues in mice. XMD8-92 could up-regulate RANKL/OPG rate, lower osteoblasts vitality by inhibiting Cyclin B1 and CDK1 expression, and promote osteoblast apoptosis by up-regulating the expression of FasL. The novel findings suggest that ERK5 activity plays an important role in maintenance of bone mass, and altered ERK5 activation in disease conditions such as osteoporosis or dexamethasone-induced osteoporosis should be considered as a potential mechanism for abnormal bone homeostasis.
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    Corylin inhibits osteoclastogenesis and attenuates postmenopausal osteoporosis in mice
    Li Xiaoqun, Xu Kaihang, Ji Fang
    2021, 25 (2):  186-190.  doi: 10.3969/j.issn.2095-4344.2974
    Abstract ( 564 )   PDF (735KB) ( 154 )   Save
    BACKGROUND: The balance of bone homeostasis is mediated by the osteoclast-related bone resorption and osteoblast-related bone formation. Over-activation of osteoclasts results in a series of bone metabolic diseases including rheumatoid arthritis and osteoporosis. The activation of nuclear factor-κB  pathway induced by receptor activator of nuclear factor-κB ligand (RANKL) plays an important role in osteoclastogenesis.
    OBJECTIVE: To explore the effect of corylin on RANKL-mediated osteoclastogenesis. 
    METHODS: RAW264.7 cells were incubated with 0, 1, 2, 4, 8, 16, 32, 64, 128 μmol/L corylin. The cytotoxicity of corylin was detected by cell counting kit-8 assay. RANKL induced the differentiation of RAW264.7 cells into osteoclasts, during which 2, 5, 10 μmol/L corylin was given. The number of osteoclasts was analyzed by TRAP staining after 5 days of intervention and the morphology and function of osteoclasts were analyzed by F-actin staining. Bone resorption assay was conducted after 2 days of intervention. The activation of nuclear factor-κB pathway was detected by western blot at 0, 15, 30, and 60 minutes of intervention. Then in vivo experiments were carried out, and the ovariectomized mice were intraperitoneally given 10 mg/kg twice a week. After 6 weeks of intervention, mouse femurs were taken for morphological analysis.
    RESULTS AND CONCLUSION: There was no cytotoxicity of corylin below the concentration of 16 μmol/L. Corylin inhibited osteoclastogenesis in a dose-dependent manner. Corylin inhibited the formation of F-actin and resorption activity of osteoclasts. Corylin inhibited RANKL-mediated nuclear factor-κB pathway. Corylin treatment reduced the bone loss in postmenopausal osteoporosis mice. Overall, corylin inhibits osteoclastogenesis via blocking nuclear factor-κB pathway and attenuates postmenopausal osteoporosis. 
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    Characteristics of bone remodeling in female ovariectomized rat models of osteoporosis undergoing Erzhi Pill extract intervention
    Li Ping, Lin Yu, Chen Xiang, Liu Zhentao, Xiao Lili, Lin Xueyi, Hua Peng
    2021, 25 (2):  191-195.  doi: 10.3969/j.issn.2095-4344.2975
    Abstract ( 389 )   PDF (935KB) ( 71 )   Save
    BACKGROUND: Some studies have shown that Erzhi Pill can improve the bone density, bone shadow area, bone mineral content and serum estradiol level of ovariectomized rats, but the possible mechanism has not been explored.
    OBJECTIVE: To investigate the effect of Erzhi Pill on bone remodeling in an ovariectomized rat model of osteoporosis.
    METHODS: A rat model of post-menopausal osteoporosis was established, and the extracts of Erzhi Pills of 6, 9, and 12 g/kg per day were administered intragastrically. Administration in each group began at the 13th week after surgery, and the samples were taken at 16, 20, and 24 weeks after surgery. The bone tissue morphology was observed by hematoxylin-eosin staining, the percentage of trabecular bone was measured by Motic 6.0 system, and the bone density of the rat right tibial bone was detected by a bone densitometer. Expressions of osteoprotegerin, nuclear factor κB receptor activating factor ligand (RANKL), tartrate-resistant acid phosphatase (TRAP) and osteocalcin mRNAs in the first lumbar vertebrae were detected by qPCR.  

    RESULTS AND CONCLUSION: The trabecular bone had a better morphological structure, and the number of trabeculae, bone miner density, and bone tissue osteoprotegerin level were significantly increased in a dose-depended manner after treatment with Erzhi Pill, whereas the mRNA levels of RANKL and TRAP decreased in a dose-depended manner after treatment with Erzhi Pill (P < 0.05). Therefore, the alcohol extract of Erzhi Pill can improve the status of high-transformation osteoporosis in ovariectomized rats, promote the expression of osteoprotegerin and inhibit the expression of RANKL, so as to inhibit the activity of osteoclasts and ultimately improve the bone remodeling in female osteoporotic rats.

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    Preparing a blunt contusion model of rabbit skeletal muscle under different blow strengths
    Liu Xing, Wei Xiaohan, Deng Jie, Li Zhongming
    2021, 25 (2):  196-200.  doi: 10.3969/j.issn.2095-4344.2977
    Abstract ( 382 )   PDF (967KB) ( 45 )   Save
    BACKGROUND: In current studies regarding skeletal muscle blunt contusion model, rats are often taken as the experimental object. The authors believe that rabbits are the better experimental object to observe the macroscopic and imaging changes after blunt contusion.  

    OBJECTIVE: Based on the self-made heavy-duty smashing instrument as the experimental basis, to replicate the skeletal muscle injury model according to different strike heights, which is expected to provide a reference basis for the subsequent replication of the rabbit gastrocnemius blunt contusion model.

    METHODS: Thirty-three New Zealand rabbits were randomly divided into four groups: normal group (n=3), 75 cm strike group (n=10), 50 cm strike group (n=10), 25 cm strike group (n=10). Except for the normal group, all strikes were performed with different strengths. The severity of skeletal muscle damage after different strikes was compared through gross observation, hematoxylin-eosin staining, Masson staining, and ultrasound imaging.  

    RESULTS AND CONCLUSION: The 50 cm height blow can cause moderate damage to the skeletal muscle, and the natural recovery time is over 21 days, which is suitable for the establishment of skeletal muscle blunt contusion model. The 75 cm height blow is likely to cause fractures, resulting in more deaths. The 25 cm height blow can cause mild damage that can be recovered within 7 days, which is not suitable for the establishment of skeletal muscle injury model.

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    Musculoaponeurotic manipulation changes contractile mechanics of the skeletal muscle in rabbits with knee osteoarthritis
    Huang Xudong, Yi Zhiyong, Han Qingmin
    2021, 25 (2):  201-204.  doi: 10.3969/j.issn.2095-4344.2972
    Abstract ( 482 )   PDF (645KB) ( 76 )   Save
    BACKGROUND: The team’s previous research found that foot-Yangming meridian and foot-Taiyang meridian play an important role in the pathogenesis of knee osteoarthritis, with the correspondence to the hamstring muscle and quadriceps muscle, respectively. Under the guidance of meridian syndrome differentiation, manipulative therapy for knee osteoarthritis has achieved good results, but its mechanism of action is not yet clear.
    OBJECTIVE: To observe the effect of aponeurotic channel manipulation on contractile mechanics of the skeletal muscle in rabbits with knee osteoarthritis. 
    METHODS: Twelve New Zealand rabbits were randomly divided into three groups, a musculoaponeurotic manipulation group, a conventional manipulation 
    group and a control group, with four rabbits in each group. The right knee arthritis model was established by the method of video right rear knee extension and fixed brake. The former two groups were treated by musculoaponeurotic manipulation and conventional manipulation respectively on the 2nd day after modeling, three times a week for 2 weeks in total. Rats in the control group were fed normally. At 1 week after the end of the treatment, the quadriceps and hamstring muscles of the right thigh were taken for contractile mechanics test. The experiment was approved by the Experimental Animal Ethics Committee of Guangzhou University of Chinese Medicine (approval No. 20190221018)
    RESULTS AND CONCLUSION: After 2 weeks of treatment, the single contraction amplitude, tetanic contraction amplitude and elastic modulus of the quadriceps femoris and hamstring muscle in the two treatment groups were better than those in the control group (P < 0.05). The amplitude of single contraction and tetanic contraction in the musculoaponeurotic manipulation group was better than that in the conventional manipulation group (P  < 0.05). These findings indicate that musculoaponeurotic manipulation can effectively improve the contractile mechanics index of rabbit knee osteoarthritis model, so as to improve the clinical symptoms and delay the progress of the disease.
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    Alcohol extract of Morinda officinalis improves lipid metabolism and bone metabolism in ovariectomized obese rats
    Huang Zhusong, Lin Yu, Chen Xiang, Lan Jinfu, Guan Yong, Gao Xi
    2021, 25 (2):  205-210.  doi: 10.3969/j.issn.2095-4344.2973
    Abstract ( 382 )   PDF (654KB) ( 90 )   Save
    BACKGROUND: Previous studies have showed that the alcohol extract of Morinda officinalis can effectively improve the bone quality and body mass of obese rats after ovariectomy. However, the exact mechanism is unclear. In this study, leptin and leptin receptor were used as the breakthrough point to investigate the effect of alcohol extract of Morinda officinalis on lipid metabolism and bone metabolism in ovariectomized obese rats. 
    OBJECTIVE: To investigate the effects of alcohol extract of Morinda officinalis lipid metabolism and bone metabolism in ovariectomized obese rats.
    METHODS: A total of 160 SPF female Sprague-Dawley rats were randomly divided into an osteoporosis group (n=120) and a sham operation group (n=40). A postmenopausal osteoporosis model was made in the osteoporosis group by removing both ovaries. After modeling, rats in the osteoporosis group were randomly subdivided into a normal diet group, a high-fat diet group and a high-fat diet + Morinda officinalis alcohol extract group, with 40 rats in each group. The sham operation group and the normal diet group were fed with ordinary diet, while the high-fat diet group and the high-fat diet + Morinda officinalis alcohol extract group were fed with high-fat diet. The high-fat diet + Morinda officinalis alcohol extract group was gavaged with 20 g/kg Morinda officinalis alcohol extract once a day, and the remaining three groups were gavaged with 2 mL of normal saline. The study protocol was approved by the Animal Ethic Committee of Fuzhou Second Hospital of Xiamen University in September 2018 with an approval No. 20180019. 
    RESULTS AND CONCLUSION: Compared with the sham operation group, serum leptin, leptin receptor, osteoprotegerin and high-density lipoprotein cholesterol levels were significantly lower in ovariectomized rats (P < 0.05), whereas osteocalcin, RANKL, tartrate-resistant acid phosphatase 5b, cholesterol, triacylglycerol, and low-density lipoprotein cholesterol levels were significantly higher in ovariectomized rats (P < 0.05). Compared with the high-fat diet group, serum leptin, leptin receptor, osteoprotegerin and high-density lipoprotein cholesterol levels were significantly increased in the high-fat diet + Morinda officinalis alcohol extract group (P < 0.05), whereas osteocalcin, RANKL, tartrate-resistant acid phosphatase 5b, cholesterol, triacylglycerol, and low-density lipoprotein cholesterol levels were decreased to different extents in the high-fat diet + Morinda officinalis alcohol extract group (P < 0.05). To conclusion, the alcohol extract of Morindus officinalis can up-regulate the leptin and leptin receptor expression in serum of ovariectomized obese rats, so as to improve the abnormal bone metabolism and lipid metabolism in ovariectomized obese rats.
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    Cornuside I inhibits interleukin-6-mediated inflammation in knee osteoarthritis rats
    Li Huye, Dou Zenghua, Kong Deyuan, Cai Jinlian, Li Zeqing
    2021, 25 (2):  211-215.  doi: 10.3969/j.issn.2095-4344.2971
    Abstract ( 442 )   PDF (799KB) ( 34 )   Save

    BACKGROUND: Studies have shown that cornuside I (Cor I) can promote the proliferation of osteoblasts and chondrocytes, but its role and mechanism in repairing early cartilage damage of knee osteoarthritis are not clear.

    OBJECTIVE: To investigate the effect of Cor I on knee osteoarthritis rats and its effect on interleukin-6 (IL-6)/JAK2/STAT3 signaling pathway. 
    METHODS: Fifty healthy male Sprague-Dawley rats were randomly divided into sham operation group, knee osteoarthritis model group (model group), Cor I high-dose group (Cor I-H), and cornuside I low-dose group (Cor I-L) and positive control group (celecoxib), with 10 rats in each group. Except the sham operation group, the knee osteoarthritis rat model was prepared according to the Hulth method. The Cor I-L and Cor I-H groups were administrated with Cor I 1.25 and 5 g/kg once a day for 6 weeks after modeling. The positive control group was fed with 4 mg/kg celecoxib, and the sham operation and model groups were fed with the same amount of normal saline once a day. The rats were sacrificed by cervical dislocation 24 hours after the last administration. Hematoxylin-eosin staining was used to observe the pathological and morphological changes of articular cartilage tissue in rats, and the degree of cartilage degeneration  was scored according to Mankin’s method. Interleukin-1β, tumor necrosis and matrix metalloproteinase-13 in rat cartilage tissue were detected by ELISA. Interleukin-6 and STAT3 protein expression in rat articular cartilage tissue were detected by immunohistochemistry. Relative expressions of JAK2, p-JAK2, STAT3, and p-STAT3 were detected by western blot. 
    RESULTS AND CONCLUSION: The articular cartilage of rats was severely damaged in the model group, where obvious chondrocyte damage and disordered arrangement of chondrocytes were observed. The joint structure of the rats in the Cor I-H, Cor I-L and positive control groups tended to be normal, the chondrocytes were distributed uniformly, and the fissures were reduced. Compared with the model group, the relevant indicators in the Cor I-L and Cor I-H groups were significantly reduced, including the Mankin’s score of cartilage tissue, interleukin-1β, tumor necrosis factor α and matrix metalloproteinase-13 levels (P < 0.05), interleukin-6 and STAT3 immunoreactivities (P < 0.01) as well as the relative expression levels of p-JAK2 and p-STAT3 and ratios of p-JAK2/JAK2 and p-STAT3/STAT3 (P < 0.05). Compared with the positive control group, these indicators mentioned above were significantly increased in the Cor I-L group (P < 0.05); however, there was no significant difference between the positive control group and Cor I-H group. Results have confirmed that Cor I can delay knee osteoarthritis-induced cartilage tissue degeneration, and its mechanism may be related to reducing the were expression of inflammatory factors and inhibiting the inflammatory response mediated by interleukin-6/JAK2/STAT3 signaling pathway.

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    Pressing local acupoints plus adjustion of the knee joint in a sitting position for treating knee osteoarthritis: a randomized controlled trial
    Xu Hui, Kang Bingxin, Zhong Sheng, Gao Chenxin, Zhao Chi, Qiu Guowei, Sun Songtao, Xie Jun, Xiao Lianbo, Shi Qi
    2021, 25 (2):  216-221.  doi: 10.3969/j.issn.2095-4344.2954
    Abstract ( 397 )   PDF (712KB) ( 124 )   Save
    BACKGROUND: Tuina for knee osteoarthritis has obvious curative effect, and has been widely used in clinical practice due to its characteristics, including simple operation, popularization, and good comfort.
    OBJECTIVE: To observe the clinical effect of the combination of pressing points of local acupoint plus adjustion of the knee joint in a sitting position in the treatment of knee osteoarthritis under “emphasizing both bones and tendons” as the guiding ideology of Tuina.
    METHODS: Ninety-one patients with knee osteoarthritis were randomly divided into a treatment group (n=46) and a control group (n=45). The treatment group was given the pressing points of local acupoint plus adjustion of the knee joint in a sitting position, and the control group was treated with oral celecoxib for 4 weeks. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, the pressure pain thresholds, the Visual Analogue Scale scores at rest and at movement, and Hospital Anxiety and Depression Scale score were analyzed before and after treatment. The total clinical efficacy and adverse reaction index score were observed. The study was approved by the Ethics Committee of Guanghua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine on June 5, 2017, approval number: 2017-k-11. All participants were informed of the trial protocol and process, and signed informed consent. This study was registered at the Chinese Clinical Trial Registry (http://www.chictr.org.cn/searchproj.aspx, ChiCTR1900022928) on May 4, 2019. 
    RESULTS AND CONCLUSION: After 4 weeks of treatment: (1) The scores of WOMAC in both groups were improved compared with the baseline, and those in the treatment group decreased more significantly (P < 0.05); (2) the pressure pain thresholds of two groups were significantly improved compared with the baseline, and the treatment group was better than the control group (P  < 0.05); (3) the Visual Analogue Scale scores at rest and at movement in the treatment group were decreased significantly compared with those in the control group (P < 0.05); (4) the scores of Hospital Anxiety and Depression Scale in terms of depression and anxiety in the treatment group were significantly lower than those in the control group (P  < 0.05); (5) the total effective rate of the treatment group was 93.18%, and the control group was 81.40%, with significant difference between the two groups (P  < 0.05); (6) there was no significant difference in the adverse reaction scores between the two groups (P  > 0.05). Overall, the above results indicate that with “emphasizing both bones and tendons” as the guiding ideology of Tuina, the clinical effect of the pressing points of local acupoint and adjustion of the knee joint in a sitting position in the treatment of knee osteoarthritis is better than that of oral celecoxib.
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    Posttraumatic progression of cartilage degeneration following anterior cruciate ligament reconstruction: a second-look arthroscopic analysis
    Lu Mingfeng, Zhao Lilian, Xing Jisi, He Lilei, Xu Ting, Wang Changbing
    2021, 25 (2):  222-227.  doi: 10.3969/j.issn.2095-4344.2960
    Abstract ( 375 )   PDF (752KB) ( 38 )   Save
    BACKGROUND: Anterior cruciate ligament (ACL) reconstruction can restore the stability of knee joint, reduce the mechanical stress on meniscus and cartilage, and avoid the occurrence and development of secondary injury including traumatic arthritis. However, some studies have shown that ACL reconstruction cannot completely prevent the occurrence and development of osteoarthritis, and significant changes in osteoarthritis can be found in the long-term imaging examination after operation. Therefore, there is still controversy about the role of ACL reconstruction in preventing secondary cartilage damage and secondary osteoarthritis, and the changes in cartilage after ACL reconstruction, especially at early stage after ACL reconstruction, have not been fully elucidated. 
    OBJECTIVE: To analyze articular cartilage changes during arthroscopic anterior cruciate ligament reconstruction and second-look arthroscopic evaluation.
    METHODS: From July 2015 to July 2016, 50 cases of ACL reconstruction with hamstring tendon and second-look arthroscopic exploration were analyzed retrospectively. The cartilage damage and healing between the ACL reconstruction and second-look exploration were recorded. At the same time, the influence of age on articular cartilage was analyzed. The study protocol was approved by the Ethics Committee of Foshan TCM Hospital with an approval No. 20160315.
    RESULTS AND CONCLUSION: No infection, graft absorption and other complications occurred in all patients. During the second-look exploration, the Lysholm score, International Knee Documentation Committee evaluation score, Tegner score and KT-1000 lateral difference were significantly improved compared with those before operation (P < 0.05). During the second-look exploration, the articular cartilage of all patients had a trend of deterioration; except for the medial tibial platform and the lateral femoral condyle, subpatellar, trochlear, medial femoral condyle and tibial lateral platform articular cartilage tissues were significantly worsened compared with those after the initial ACL reconstruction (P < 0.05). There was no significant difference in the condition of articular cartilage between the patients under 30 years and over 30 years old in the initial ACL reconstruction (P > 0.05). During the second-look exploration, the patients with anterior cruciate ligament reconstruction who were 30 years old and above had significantly worse subpatellar, trochlear, medial femoral condyle, lateral femoral condyle and tibial lateral platform cartilage tissues than those under 30 years (P < 0.05). The results suggest that the reconstruction of ACL with autogenous hamstring tendon can achieve better stability and function of the knee joint. However, even in the early postoperative period, osteoarthritis is still progressing. Therefore, it is necessary for patients with ACL reconstruction to pay attention to the progression of osteoarthritis and the appearance of knee joint symptoms.
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    Immediate effects of extracorporeal shock wave on muscle tone, stiffness, and elasticity of wrist flexor in stroke patients
    Bao Sairong, Zhang Qiming, Yang Xingping, Liu Chunlong
    2021, 25 (2):  228-231.  doi: 10.3969/j.issn.2095-4344.2874
    Abstract ( 550 )   PDF (613KB) ( 110 )   Save

    BACKGROUND: Studies have shown that the elasticity of the radial wrist flexor and ulnar wrist flexor muscles is related to the power of gripping, and the tension and stiffness directly affect the pinching force on the thumb side and the palm gripping ability. Extracorporeal shock waves have a definite effect on alleviating post-stroke limb spasm.

    OBJECTIVE: To observe the immediate effect of extracorporeal shock wave therapy on muscle tone, stiffness, and elasticity of flexor carpi radialis and flexor carpi ulnaris in stroke patients.
    METHODS: Twenty stroke patients with hemiplegia who met the inclusion criteria were given a single session of extracorporeal shock wave therapy on flexor carpi radialis and flexor carpi ulnaris. The oscillation frequency, logarithmic decrement, dynamic stiffness of flexor carpi radialis and flexor carpi ulnaris were detected using a muscle tone measurement device Myoton-3 before treatment and 5 minutes after treatment. At the same time, the Modified Tardieu Scale was used to evaluate spasticity of wrist flexor before and after treatment. The study protocol was implemented in accordance with the relevant ethical requirements of the First Affiliated Hospital of Guangdong Pharmaceutical University, and the enrolled patients and their families were fully informed of the whole trial.
    RESULTS AND CONCLUSION: After treatment, the oscillation frequency, logarithmic decrement, dynamic stiffness, and quality of muscle response of flexor carpi radialis and flexor carpi ulnaris were significantly reduced in all the patients (P < 0.01). The maximum passive range of motion of wrist extension and angle of catch following a fast velocity stretch wrist flexor were significantly higher than those before (P < 0.01). In conclusion, a single session of extracorporeal shock wave therapy can effectively reduce muscle tone and stiffness, and improve elasticity of flexor carpi radialis and flexor carpi ulnaris in stroke patients. And it can also improve the maximum passive range of motion of wrist extension. 

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    Dominant and non-dominant side knee isokinetic characteristics of Chinese calisthenics athletes
    Li Yu, Huang Peng, Wang Hong, Wang Anli
    2021, 25 (2):  232-236.  doi: 10.3969/j.issn.2095-4344.2978
    Abstract ( 479 )   PDF (621KB) ( 70 )   Save
    BACKGROUND: A comprehensive evaluation of knee muscle strength, knee hamstrings-to-quadriceps (H/Q) force ratio, and symmetry of both knee joints is beneficial to preventing knee joint injuries.  

    OBJECTIVE: To analyze the dominant and non-dominant side isokinetic characteristics of the knee joints of Chinese calisthenics athletes and bilateral symmetry of muscle strength.

    METHODS: Knee joints of 22 Chinese calisthenics athletes (aerobics group, n=8; athletics group, n=14) from Chinese national aerobics team were measured using IsoMed2000 at 60 and 180 (°)/s concentric angular speed.  
    RESULTS AND CONCLUSION: The peak torque (PT) and the relative peak torque (PT/BW) of the flexors and extensors of the knee joint in the two groups were significantly decreased with the increase of the angular speed (P < 0.001), and the H/Q ratio significantly increased with the increase of the angular speed (P < 0.001). The dominant leg flexors strength were significantly higher than that of the non-dominant leg (P < 0.05), and extensors strength and H/Q of the dominant leg were significantly higher than those of the non-dominant leg at the speed of 60 (°)/s (P < 0.05). The H/Q of the non-dominant knee joint in the aerobic group was significantly higher than that in the athletics group at the speed of 180 (°)/s (P < 0.05). In the aerobic group, the flexor peak torque of the dominant leg was significantly higher than that of the non-dominant leg at the speed of 60 (°)/s and 180 (°)/s (P < 0.05), but there was no significant difference in both side of extensors. In the athletics group, the flexor peak torque of the dominant leg was significantly higher than that of the non-dominant leg (P < 0.05), the extensors peak torque [60 (°)/s], PT/BW [60 (°)/s] and H/Q [180 (°)/s] of the dominant leg were significantly higher than those of the non-dominant leg (P < 0.05). These findings indicate that the muscle force of the dominant and non-dominant knee joints were asymmetric, and the athletes ought to improve the muscle power of the non-dominant knee joint. A lower H/Q indicates imbalance between the flexor and extensor strength of the knee joint, and it is necessary to strengthen the knee flexor strength training, especially the training for fast strength.

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    Establishment of an acute radioactive skin injury model in Wistar rats
    Chen Yutong, Li Chenchen, Liu Yang, Zheng Yaqin, Yang Xihua, An Meiwen
    2021, 25 (2):  237-241.  doi: 10.3969/j.issn.2095-4344.2981
    Abstract ( 597 )   PDF (943KB) ( 171 )   Save
    BACKGROUND: Nowadays, the ray types, animal species, irradiation modes and sites used in the establishment of animal models of radioactive skin injury in China are not consistent. Meanwhile, there is no uniform standard for the prevention and treatment of radioactive skin injury in clinical practice. 
    OBJECTIVE: To establish an ideal rat model of acute radioactive skin injury.
    METHODS: Sixty Wistar rats were randomly divided into 32, 38, 45 Gy X-ray groups (n=18) and non-irradiated group (n=6). Three X-ray irradiated groups (32, 38, 45 Gy) received single irradiation of the right posterior buttock, 300 cGy/min, 100 cm between the skin and irradiated source, for 10.67, 12.67, and 15 minutes respectively. No irradiation was given in the non-irradiated group. The study protocol was approved by the Animal Ethic Committee of Shanxi Cancer Hospital (approval No. GDY2018001).
    RESULTS AND CONCLUSION: There was no accidental death after irradiation. The body mass of the rats decreased within 3 days after irradiation, and then increased. Irradiated wound was severest at about 15 days after irradiation, and the body mass dropped again, and returned to normal 2 days later. Two weeks after radiation, with the increase of X-ray dose, the structures of rat’s skin appendages were destroyed and a large number of inflammatory cells were infiltrated, indicating that the acute radiation skin injury was dose-dependent within a certain range. On the other hand, with the increase of irradiation time, the skin wound in the 38 and 45 Gy groups gradually deepened. At the same dose, the severity of acute radiation skin injury was also positively correlated with the irradiation time. After 6 hours to 15 days of 38 Gy irradiation on the rat skin, macrophages were activated, and the expression of Nod-like receptor pyrin domain-containing protein 3 was enhanced, indicating obvious inflammatory response, and thereby verifying the reliability of the model. To conclude, it is an ideal animal model of acute X-ray skin injury model made by the X-ray linear accelerator, which is easily observed and obtained, with obvious skin inflammation expression. This model is also of high safety and strong tolerance.
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    Chemokine CCL21 in anterior cingulate cortex is involved in chronic neuropathic pain in a rat model
    Zhi Yibo, Zhang Jie, Liu Jian, Li Weiyan
    2021, 25 (2):  242-246.  doi: 10.3969/j.issn.2095-4344.2982
    Abstract ( 330 )   PDF (766KB) ( 93 )   Save
    BACKGROUND: The pathogenesis of chronic pathological pain is yet unknown. Some studies have shown that after spinal cord injury, CCL21 can activate microglia in the central nervous system and is expressed only in damaged neurons, promoting the formation of chronic pathological pain. 
    OBJECTIVE: To investigate whether the anterior cingulate cortex is involved in the formation of chronic pathological pain after inferior orbital nerve ligation in rats, and whether blocking chemokine CCL21 in the anterior cingulate cortex can reduce the chronic neuropathic pain. 
    METHODS: A total of 80 male Sprague-Dawley rats were randomly divided into 4 groups with 20 rats in each group. In the sham group, only the infraorbital nerve of the rats was exposed; in the model group, the left infraorbital nerve was ligated; in the anti-CCL21 group, CCL21 neutralizing antibodies was administered to the anterior cingulate cortex of the rats on the 7th day after surgery; and in the PBS control group, PBS solution was given into the anterior cingulate cortex of rats on the 7th day after surgery. Rats in the sham and model groups were subjected to behavioral tests on the 3rd, 5th, 7th, and 14th days after surgery, and those in the anti-CCL21 and PBS control groups were subjected to the behavioral test at 6 hours after administration. All rats were sacrificed under anesthesia after behavioral tests. The cortical tissues were taken from the anterior cingulate, and the protein content of CCL21 was determined by western blot and immunofluorescence. 
    RESULTS AND CONCLUSION: The pain threshold of the rats in the model group was lower than that in the sham group, and the expression of CCL21 in the anterior cingulate cortex was significantly higher in the model group than the sham group. After the administration of CCL21 neutralizing antibody, the expression of CCL21 was reduced to some extents, and the rat pain threshold was increased accordingly. These findings reveal that the anterior cingulate cortex of rats may be involved in the production of chronic pathological pain, and the administration of CCL21 neutralizing antibody can relief the pain.
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    Mechanism of myocardial oxidative damage in a rat model of one-time exhaustive exercise
    Xie Wenjie, Zhou Gang, Xie Jinmei, Liu Jiao, Li Pengfei, Yang Fan, Cui Di
    2021, 25 (2):  247-252.  doi: 10.3969/j.issn.2095-4344.2979
    Abstract ( 314 )   PDF (771KB) ( 41 )   Save
    BACKGROUND: Exhaustive exercise is a vigorous physical activity performed by an organism beyond its physiological limits, and it causes a series of histological changes in the body. Myocardial exercise-induced oxidative stress injury means that the organism generates free radicals through oxidative stress signal pathway to damage myocardial cells under exhaustive exercise.
    OBJECTIVE: To explore the mechanism of oxidative stress injury in rat myocardium caused by one-time exhaustive exercise based on the protein kinase C (PKC)/NOX pathway. 
    METHODS: Thirty adult male Sprague-Dawley rats were randomly divided into a control group, an exhaustive exercise group, and an exhaustive exercise+drug group, with 10 rats in each group. The exhaustive exercise+drug group was injected with PKC inhibitor chelerythrine (5 mg/kg body weight) for 3 consecutive days. Rats in the two exercise groups exercised at a speed of 25 m/min on a 0° incline treadmill until exhaustion. Immediately after exercise, blood sample was collected from each rat, and then the rat’s left ventricle was removed for hematoxylin-eosin staining to observe the morphological changes of myocardial cells. Serum and myocardial malondialdehyde and myocardial reactive oxygen species levels were detected. The protein expressions of PKC, NOX2, NOX4 and 3-NT in rat myocardial tissue were determined by western blot. 
    RESULTS AND CONCLUSION: The myocardial tissues in the exhaustive exercise and exhaustive exercise+drug groups were damaged, but the damage was significantly eased in the exhaustive exercise+drug group compared with the exhaustive exercise group. Compared with the control group, the reactive oxygen species level in the myocardial tissue of the exhaustive exercise group increased significantly (P < 0.05). Compared with the control group, the content of malondialdehyde in the myocardial tissues of the exhaustive exercise and exhaustive exercise+drug groups was significantly increased (P < 0.01), and the concentration of serum malondialdehyde in the exhaustive exercise group was significantly increased (P < 0.05). Compared with the control group, the expression levels of PKC, NOX2, NOX4 and 3-NT proteins in the myocardium of rats after exhaustive exercise were significantly increased (P < 0.01). Compared with the exhaustive exercise group, the expression levels of NOX2 and NOX4 in the myocardial tissue of rats significantly decreased in the exhaustive exercise+drug group (P < 0.01), and the expression of 3-NT significantly decreased (P < 0.05). Therefore, one-time exhaustive exercise can activate PKC and increase its protein expression in rat myocardial cells, which in turn induces an increase in the expression of NOX2 and NOX4 proteins in the myocardium, catalyzes the generation of large amounts of reactive oxygen species, and leads to the excessive production of peroxynitrite anions, thereby causing oxidative damage to the myocardium.
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    Effects of ultrashort wave on inflammatory response in rats with acute lung injury
    Liao Yuan, Qu Mengjian, Liu Jing, Zhong Peirui, Zeng Yahua, Wang Ting, Xiao Hao, Li Lan, Liu Danni, Yang Lu, Zhou Jun
    2021, 25 (2):  253-257.  doi: 10.3969/j.issn.2095-4344.2953
    Abstract ( 540 )   PDF (722KB) ( 50 )   Save
    BACKGROUND:  Ultrashort wave can inhibit the inflammatory response and is often used in symptomatic treatment of pulmonary infection. Uncontrolled inflammatory response is an important pathogenesis of acute lung injury. Inhibiting inflammation is an important strategy for controlling acute lung injury.
    OBJECTIVE: To observe the effects of ultrashort wave on inflammatory response and the expression of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in rats with acute lung injury.
    METHODS: Twenty-four 3-month-old male Sprague-Dawley rats were randomly divided into control group, acute lung injury group (model group) and ultrashort wave group (n=8 per group). Rats in the acute lung injury group and the ultrashort wave group were given intratracheal lipopolysaccharide to replicate the acute lung injury model. Rats in the control group were given intratracheal infusion of the same amount of normal saline. In the ultrashort wave group, rats were given ultrashort wave intervention immediately, 4 and 8 hours after lipopolysaccharide infusion, once for 15 minutes. Experimental animals were sacrificed 24 hours after intratracheal infusion of lipopolysaccharide or normal saline. The lung tissues of rats were compared by hematoxylin-eosin staining, lung histopathological semi-quantitative score and lung wet/dry weight ratio (W/D). Serum levels of inflammatory factors TNF-α and IL-1β were detected by ELISA, and mRNA and protein expressions of TNF-α and IL-1β were detected by RT-PCR and western blot, respectively. The study protocol was approved by the First Affiliated Hospital of Nanhua University, approved No.202002100009.
    RESULTS AND CONCLUSION: Lung W/D ratio in the acute lung injury group was significantly higher than that in the control group (P < 0.01), whereas the ratio in the ultrashort wave group was lower than that in the acute lung injury group, but there was no significant difference between the two groups. Pathological manifestations: In the model group, the lung tissue structure was obviously damaged, with different sizes of alveolar septa, the alveolar wall was incomplete, thickened and fractured, a large number of neutrophils were seen in the lung interstitium, and some red blood cells were exudated. In the ultrashort wave group, compared with the model group, the alveolar structure was relatively complete and clear, and the infiltration of inflammatory cells and red blood cell exudation from the lung interstitium were reduced. Semi-quantitative histopathological score of the lung was significantly higher in the model group than the control group (P < 0.01), but significantly reduced in the ultrashort wave group compared with the model group (P < 0.01). Serum TNF-α and IL-1β levels in the model group were significantly higher than those in the control group (both P < 0.01). After ultrashort wave exposure, the levels of serum TNF-α and IL-1β significantly decreased compared to the model group (both P < 0.05).  After ultrashort wave exposure, the mRNA and protein expressions of TNF-α and IL-1β significantly decreased compared to the model group (P < 0.01 or P < 0.05). To conclude, ultrashort wave may inhibit the inflammatory response of the lung tissue in rats with acute lung injury by down-regulating the expression of inflammatory cytokines, TNF-α and IL-1β. 
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    Chemerin, a pro-inflammatory adipokine, regulates chondrocyte proliferation and metabolism by increasing production of nitric oxide
    Yu Chengshuai, Du Gang, Pang Shenning, Lao Shan
    2021, 25 (2):  258-263.  doi: 10.3969/j.issn.2095-4344.2976
    Abstract ( 344 )   PDF (998KB) ( 37 )   Save
    BACKGROUND: The key pathological characteristics of osteoarthritis are manifested in the degeneration of the cartilage caused by inflammation, and chondrocytes are the only cells in cartilage tissues. Studies have shown that Chemerin can stimulate the migration of leukocytes to the inflammation site and increase the inflammation signal of chondrocytes, suggesting that Chemerin can play a role in arthritis. Our previous research indicated that the serum Chemerin level in patients with osteoarthritis was significantly increased, and the Chemerin level in the synovial fluid was related to the severity of osteoarthritis based on the Kellgren-Lawrence classification. Chemerin may be used as an inflammatory factor in osteoarthritis.
    OBJECTIVE: To investigate the effect of Chemerin on the proliferation and metabolism of chondrocytes.
    METHODS: The chondrocytes from neonatal mice were isolated by collagenase type II digestion, and then cultured. Cell growth curves were established and the range of concentrations of Chemerin that exhibited toxicity to normal chondrocytes was screened using an MTT assay. Subsequently, 10 μg/L interleukin-1β was used to stimulate the chondrocytes in order to establish an in vitro model of osteoarthritis induction. After the chondrocytes had been cultured in the presence of the drug for 2 days, cell morphology, proliferation and metabolism were evaluated by hematoxylin-eosin staining and diacetate fluorescein/propidium iodide staining. In addition, the expression of inducible nitric oxide polymerase was analyzed by measuring the secretion of nitric oxide. Furthermore, qRT-PCR was used to quantify mRNA expression of proteoglycan, type II collagen α1, matrix metalloprotease-13 and nitric oxide synthase 2.  
    RESULTS AND CONCLUSION: The chondrocytes cultured in vitro exhibited healthy activity and morphology. Furthermore, chemerin (50 μg/L) and interleukin-1β (10 μg/L) were able to reduce the synthesis of extracellular matrix, enhance the secretion of nitric oxide and increase chondrocyte apoptosis. More importantly, the qRT-PCR results indicated that Chemerin and interleukin-1β caused similar effects, by which the expression of cartilage-specific genes was downregulated and catabolism-related genes upregulated. As a pro-inflammatory factor, Chemerin can increase the generation of nitric oxide in chondrocytes, regulate cell metabolism, stimulate cell apoptosis and act synergistically with interleukin-1β.
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    Pheretima extract ameliorates nucleus pulposus cell degeneration in the intervertebral disc by inhibiting nuclear factor-kappa B pathway 
    Fu Yuanfei, He Shenghua, Lai Juyi, Sun Zhitao, Feng Hualong, Lan Zhiming
    2021, 25 (2):  264-268.  doi: 10.3969/j.issn.2095-4344.2983
    Abstract ( 348 )   PDF (777KB) ( 87 )   Save
    BACKGROUND: Inflammatory microenvironment of nucleus pulposus cells is an important factor to promote nucleus pulposus degeneration. Inflammatory factors amplify the cascade of inflammation by activating the nuclear factor-κB signaling pathway to form a vicious cycle and accelerate the process of nucleus pulposus degeneration. It could delay the progression of intervertebral disc degeneration by inhibiting the activity of nuclear factor-κB signaling pathway.  
    OBJECTIVE: To investigate the mechanism of Pheretima extract on nucleus pulposus cell degeneration.
    METHODS: Pheretima active ingredients were extracted by cold dipping. The cell counting kit-8 method was used to detect the effects of 0, 25, 50, 100, 200, 400 mg/L Pheretima extract on rat nucleus pulposus cell activity, and then suitable drug concentrations were chosen for following research. Tumor necrosis factor-α (TNF-α) was used to stimulate nucleus pulposus cells to induce the nucleus pulposus cell degeneration model. Pheretima extracts (50, 100, 200 mg/L) were used to treat the cell model. Western blot was used to detect the expression of Aggrecan, Collagen II, TNF-α and interleukin-6. RT-qPCR was used to detect mRNA expression of TNF-α and interleukin-6. Immunofluorescence was used to detect the nuclear expression of P65 in the nucleus pulposus cells.  
    RESULTS AND CONCLUSION: Compared with the blank group (0 mg/L), 400 mg/L Pheretima extract inhibited the activity of nucleus pulposus cells, and 50, 100, 200 mg/L extracts of Pheretima were used for subsequent experimental research. Compared with the normal group, the expression of Aggrecan and Collagen II in the model group was reduced, the protein and mRNA expression levels of TNF-α and interleukin-6 were up-regulated, and P65 expression increased in the nucleus. Compared with the model group, Pheretima extract could increase the expression of Aggrecan and Collagen II, down-regulate the protein and mRNA levels of TNF-α and interleukin-6, and reduce P65 expression in the nucleus. To conclude, Pheretima extract can reduce the expression of inflammatory factors, increase the synthesis of extracellular matrix and inhibit nuclear factor-κB pathway activity. Pheretima extract may ameliorate nucleus pulposus cell degeneration in the intervertebral disc via the nuclear factor-κB signaling pathway.
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    Increased FoxO1 DNA methylation level in homocysteine-induced podocyte apoptosis
    Liu Kun, Xie Lin, Cao Jun, Ding Ning, Xu Lingbo, Ma Shengchao, Li Guizhong , Jiang Yideng, Lu Guanjun
    2021, 25 (2):  269-273.  doi: 10.3969/j.issn.2095-4344.2980
    Abstract ( 393 )   PDF (761KB) ( 36 )   Save
    BACKGROUND: The increase of homocysteine can lead to renal injury and podocyte apoptosis, but the specific mechanism is not clear. 
    OBJECTIVE: To investigate the effect of Forkhead box O1 (FoxO1) and its DNA methylation in podocyte apoptosis induced by homocysteine. 
    METHODS: Mouse renal podocytes (MPC-5) were cultured in vitro and divided into control group (0 μmol/L homocysteine) and homocysteine group (80 μmol/L homocysteine). After 48 hours of intervention, the expression of podocyte apoptosis-related proteins Bax, caspase12 and Bcl-2 was detected by immunofluorescence technique; the expression level of FoxO1 mRNA was detected by real-time fluorescence quantitative PCR; the protein expression levels of FoxO1 and DNMT1 were detected by western blot; DNA methylation level of FoxO1 was detected by nested methylation-specific PCR.
    RESULTS AND CONCLUSION: Compared with the control group, the expression levels of Bax and caspase12 protein in podocytes of the homocysteine group were significantly increased, while the expression of Bcl-2 protein was significantly decreased. The expression levels of FoxO1 mRNA and protein were significantly decreased in the homocysteine group compared with the control group (P < 0.01). At the same time, the methylation level of FoxO1 DNA in the homocysteine group was significantly higher than that in the control group (P < 0.01), and the expression of DNMT1 protein in podocytes in the homocysteine group was significantly higher than that in the control group (P < 0.01). To conclude, FoxO1 DNA hypermethylation plays a significant role in podocyte apoptosis induced by homocysteine, whereas DNMT1 participates in homocysteine-induced podocyte apoptosis. 
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    Maxing Xiongting Mixture regulates factors relevant to lung reshaping and vascular remodeling of hypoxic pulmonary hypertension rats
    Li Songtao, Li Xinyi, Song Yunfeng, Ning Jiayin, Ren Qiang, Yang Renxu, Peng Bo
    2021, 25 (2):  274-280.  doi: 10.3969/j.issn.2095-4344.2952
    Abstract ( 326 )   PDF (1033KB) ( 92 )   Save
    BACKGROUND: Hypoxic pulmonary hypertension is a key link in the progression from chronic obstructive pulmonary disease to cor pulmonale. Its severity is closely related to disease development and prognosis. Current treatments cannot prevent or reverse disease progression. Maxing Xiongting Mixture has significant effect on hypoxic pulmonary hypertension with the syndrome of intermingled phlegm and blood stasis.
    OBJECTIVE: To study how the Maxing Xiongting Mixture regulates relevant factors of lung reshaping and vascular remodeling of hypoxic pulmonary hypertension rats with the syndrome of intermingled phlegm and blood stasis. 
    METHODS: Seventy Sprague-Dawley rats, 5 weeks old, were randomly divided into normal group (n=10) and model group (n=60), where acute cor pulmonale model was prepared by injecting 50 mg/kg monocrotaline solution (1%) intraperitoneally, followed by forced smoking and swimming 6 days a week lasting for 4 weeks. Except for 10 rats in the normal group, there were 46 model rats in the model group. According to the normal distribution of body mass, 40 rats were selected and randomly divided into 4 groups: model group, high-dose Maxing Xiongting Mixture group (MH), low-dose Maxing Xiongting Mixture group (ML) and fasudil group, with 10 rats in each group. Rats in MH and ML groups were respectively given Maxing Xiongting Mixture at 20 g/(kg·d) and 5 g/(kg·d), respectively and those in the fasudil group were given fasudil at a dose of 10 mg/(kg·d). Other groups were given equal amount of saline. Administration was given intraperitoneally and intragastrically, once a day for 14 days in total. RT-PCR was used to test the expression of factors related to lung reshaping and vascular remodeling, including RhoA, stromelysin 1 and tumor necrosis factor-α mRNAs. An approval for the study was obtained from the Ethics Committee of Chengdu University of Traditional Chinese Medicine (approval No. 2017-03).
    RESULTS AND CONCLUSION: Compared with the model group, the expressions of RhoA, stromelysin 1, and tumor necrosis factor-α mRNAs were significantly lowered in the MH group (all P < 0.05); the mRNA expressions of RhoA (P < 0.05), stromelysin 1 (P < 0.01) and tumor necrosis factor-α (P < 0.05) were significantly reduced in the fasudil group; and no significant changes in the above expressions of relevant factors were found in the ML group (P > 0.05). To conclude, Maxing Xiongting Mixture, which is similar to fasudil, intervenes lung reshaping and vascular remodeling of hypoxic pulmonary hypertension rats with the syndrome of intermingled phlegm and blood stasis by inhibiting the expressions of RhoA, stromelysin 1, and tumor necrosis factor-α mRNAs. 
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    Development and evaluation of alpha-galactosyl antigen-deficient rabbit model
    Mu Yufeng, Wei Lina, Wu Yong, Shao Anliang, Chen Liang, Qu Shuxin, Xu Liming
    2021, 25 (2):  281-285.  doi: 10.3969/j.issn.2095-4344.2958
    Abstract ( 377 )   PDF (728KB) ( 44 )   Save

    BACKGROUND: α-Galactosyl (α-Gal) is the main target antigen in hyperacute rejection resulting from animal tissues or xenotransplantation. 
    OBJECTIVE: To develop a Gal antigen-deficient rabbit model in order to objectively evaluate the immunogenicity risk of animal-derived biomaterials and the local response to implantation in the host. 
    METHODS: New Zealand white rabbits, SPF grade, 6-8 months old, were selected as model animals to prepare Gal antigen-deficient rabbits. Using CRISPR/Cas9-mediated gene editing technology, two complementary sgRNAs were designed and constructed for the 8th exon of GGTA1 gene to regulate Gal antigen expression in rabbits. After transcription, GGTA1 sgRNA mRNAs and Cas9 mRNA were co-microinjected into in vitro cultured rabbit fertilized eggs, which were then implanted into surrogate mother rabbits after a brief in vitro culture, and the neonatal rabbits were obtained by natural pregnancy. The success of gene editing was verified by gel electrophoresis and gene sequencing. The expression of Gal antigen was detected with reference to the method given by the industry standard (YY/T 1561-2017). The study protocol was approved by the Animal Ethics Committee of the Laboratory Animal Resources Laboratory of the National Institutes for Food and Drug Control (approval No. 2017(B)007).
    RESULTS AND CONCLUSION: The embryos were transferred to four surrogate rabbits after gene editing. FIfteen gene-edited pups were obtained after natural pregnancy. The appearance and feeding behavior of the pups were not abnormal. Gel electrophoresis and gene sequencing results showed that 14 out of 15 rabbits were successfully edited, but the edited bases were not the same. The Gal antigen of the main organs was detected randomly, and its expression was reduced by more than 99.96%. Therefore, Gal-antigen-deficient rabbits are expected to be used for immunogenic risk assessment of animal-derived medical devices and in situ implantation experiments of heterogeneous bone and cornea, in order to be able to more objectively and scientifically evaluate the safety and effectiveness of animal-derived medical devices.

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    Effects of baicalin on oxidative stress in BEAS-2B cells stimulated by lipopolysaccharide combined with adenosine triphosphate
    Cong Renyuan, Yuan Jing, Xia Jinchan, Sun Ying
    2021, 25 (2):  286-291.  doi: 10.3969/j.issn.2095-4344.2962
    Abstract ( 410 )   PDF (649KB) ( 85 )   Save
    BACKGROUND: Oxidative stress and inflammatory reaction play important roles in the occurrence and development of acute lung injury. Studies have shown that baicalin has biological activities such as antioxidant and anti-inflammatory. 
    OBJECTIVE: To investigate the effects of baicalin on oxidative stress and TXNIP/NLRP3 pathway in BEAS-2B cells induced by lipopolysaccharide combined with adenosine triphosphate. 
    METHODS: The inflammatory model of BEAS-2B cells was established by lipopolysaccharide combined with adenosine triphosphate stimulation. The cells were randomly divided into blank control group, model group and baicalin group (2.5, 5, and 10 mg/L). MTT method was used to detect the cell vitality. ELISA was used to detect the levels of interleukin-1β, interleukin-6, interleukin-18 and tumor necrosis factor-ɑ. The changes of intracellular reactive oxygen species, superoxide dismutase and malondialdehyde were measured by fluorescent probe DCFH-DA and microplate reader, respectively. qRT-PCR was used to detect the mRNA expression levels of TXNIP, NLRP3, ASC, Caspase-1, interleukin-1β and interleukin-18. Western blot was used to detect the protein expression of TXNIP, Caspase-1 and NLRP3 in the cells. 
    RESULTS AND CONCLUSION: Compared with the blank control group, intracellular reactive oxygen species and malondialdehyde levels were significantly increased in the model group, while the activity of superoxide dismutase and cell viability were significantly decreased. There was a significant increase in the secretion levels of interleukin-1β, interleukin-6, interleukin-18, tumor necrosis factor-ɑ as well as the gene expression levels of TXNIP, NLRP3, ASC, Caspase-1, interleukin-1β, interleukin-18 and the protein expression levels of TXNIP, Caspase-1 and NLRP3 in the model group compared with the blank control group. Baicalin could considerably reverse the above indicators in a concentration-dependent manner. To conclude, baicalin has a protective effect on the damage in BEAS-2B cells stimulated by lipopolysaccharide combined with adenosine triphosphate, by reducing oxidative stress and inhibiting the activation of the TXNIP/NLRP3 signaling pathway. 
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    Animal models of osteonecrosis of the femoral head: modeling methods and characteristics
    Wang Yan, Dong Benchao, Wang Ying, Sun Lei, Lu Bin, Bai Haohao, Tian Aixian, Ma Jianxiong, Ma Xinlong
    2021, 25 (2):  292-297.  doi: 10.3969/j.issn.2095-4344.2961
    Abstract ( 553 )   PDF (594KB) ( 50 )   Save
    BACKGROUND: Osteonecrosis of the femoral head is a common and difficult disease in orthopedic department, with a high disability rate. At present, there is still no clear conclusion about its pathogenesis, as well as no ideal way to cure the disease. Therefore, it is necessary to use animal models to further study the etiology, pathogenesis and treatment of the disease.
    OBJECTIVE: To review some common animal models of osteonecrosis of the femoral head in recent years, summarize and analyze the corresponding clinical symptoms and the advantages and disadvantages of each animal model, so as to provide the basis for the follow-up research in this area.
    METHODS: The keywords of “femoral head necrosis” and “animal model” were used to search articles published between January 2000 to December 2019 in WanFang, VIP, CNKI, PubMed, Embase and Medline databases. Finally 42 eligible articles were included.
    RESULTS AND CONCLUSION: The animal model of osteonecrosis of the femoral head has been used for many years, which really improves the understanding and research of this kind of orthopedic disease. However, at present, the related research of such diseases still needs to be further explored, through the establishment of more ideal and perfect animal models to achieve a new breakthrough in this research field.
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    Research strategy of gene editing technology in the gene treatment of osteoarthritis 
    Ma Dujun, Peng Liping, Chen Feng, Jiang Shunwan, Jiang Jinting, Gao Kun, Lin Zhanpeng
    2021, 25 (2):  298-303.  doi: 10.3969/j.issn.2095-4344.2956
    Abstract ( 436 )   PDF (794KB) ( 61 )   Save
    BACKGROUND: Osteoarthritis (OA) is a common chronic disease in middle-aged and elderly people with cartilage degeneration injury as the main pathological change. Physiotherapy and drugs can be used to treat OA in the early stage, and surgery can be used in the middle and late stage, but the overall treatment effect is poor.
    OBJECTIVE: To review the latest development of gene editing technology in OA gene research worldwide.
    METHODS: With “Gene editing, Osteoarthritis, The CRISPR/Cas9 technology, Gene therapy, Stem cells” as keywords in English and Chinese, respectively, literatures related to OA gene therapy and gene editing technology were retrieved from PubMed, CNKI and WanFang databases from 2000 to 2020, and summarized and analyzed.
    RESULTS AND CONCLUSION: A variety of growth factors and regulatory factors are beneficial to promoting the synthesis of cartilage matrix and reducing joint inflammation, thereby providing a seed source for gene targeted modification. Gene editing technology can be used to introduce the target genes into the body by oral, surgical, topical, and articular injection through genetic modification of the congenital embryo or release of biological agents to specific anatomical targets to play the role of gene therapy. New progress has been made in the modification and application of OA disease genes. Gene editing has taken a new direction in its application. However, more animal experiments and clinical trials are needed in the future to study the tissue engineering of genetic modification of OA disease genes.
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    Self-renewal and signal regulation of myosatellite cells: application hotspots and problems
    Li Lunyu, Jin Songlin, Huang Zenghao, Kang Liang, Hu Yushi, Ding Haili
    2021, 25 (2):  304-310.  doi: 10.3969/j.issn.2095-4344.2959
    Abstract ( 628 )   PDF (750KB) ( 196 )   Save
    BACKGROUND: The self-renewal of muscle satellite cells is an important part of the regeneration and repair of the skeletal muscle after muscle fiber damage. 
    OBJECTIVE: To review the research progress of self-renewal and related signal regulation of myosatellite cells. 
    METHODS: The relevant literatures were searched in PubMed and Web of Science database, using the keywords of “skeletal muscle, satellite cells, stem cell, muscle repair” and “signal pathway, self-renewal.” After reading titles and abstracts, the literatures that were not related to the theme of the article were excluded and finally, 60 literatures were included for result analysis. 
    RESULTS AND CONCLUSION: In recent years, increasing attention has been paid to the involvement of muscle satellite cells in the adaptive process of skeletal muscle, which are widely and rapidly activated to generate muscle cells to act on skeletal muscle injury. The diversity and complexity of the self-renewal mechanism of muscle satellite cells can affect the repair and regeneration of skeletal muscle by regulating the differentiation of muscle satellite cells. At present, a large number of research on the self-renewal of muscle satellite cells are related to aging. A good application prospect is displayed by improving the activation degree of muscle satellite cells, increasing the number of individuals involved, and exploring their participation in  tissue regeneration and repair, but more in-depth research and exploration are needed to clarify the specific mechanism.
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    Cell autophagy, pathway, regulation and its multiple correlations with pulmonary hypertension
    Chen Xinling, Wang Shenglan
    2021, 25 (2):  311-316.  doi: 10.3969/j.issn.2095-4344.2951
    Abstract ( 413 )   PDF (777KB) ( 121 )   Save
    BACKGROUND:  Autophagy is a highly conserved lysosome degradation pathway in eukaryotic cells, which is mainly used to degrade intracellular long-lived proteins, damaged organelles, invasive pathogenic microorganisms, and so on. Recent studies have shown that cell autophagy plays an important role in the occurrence and development of pulmonary hypertension, so elucidating the molecular mechanism of cell autophagy is particularly important for the treatment of pulmonary hypertension.
    OBJECTIVE: To generalize the process of autophagy and the regulation of cell signaling pathways on autophagy, to summarize the effect of autophagy in pulmonary hypertension, and to put forward methods to study cell autophagy.
    METHODS: With the search formula “Cell Autophagy and Cell Signaling Pathway”, CNKI, WanFang, and VIP were retrieved, and with the search formula “(“ Autophagy “[Mesh]) AND” Signal Transduction “[Mesh]” and “(“Hypertension, Pulmonary”[Mesh]) AND “Autophagy”[Mesh],’’ PubMed was searched. After preliminary screening of the title and abstract to exclude stale and poor-quality literature, a total of 44 literatures were included for analysis.
    RESULTS AND CONCLUSION: In recent years, studies have found that autophagy is related to many diseases, and it is unknown about whether autophagy has protective or harmful effects. Many signaling pathways have participated in and regulated autophagy. This paper clarifies the effect of cell signaling pathways on autophagy; explores the relationship between cell signaling pathway proteins and the upstream and downstream of cell autophagy proteins; elucidates the influence of a substance on cell autophagy and signaling pathway, and  the substance maybe regulates cell autophagy via a certain pathway. 
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    Review of interferon alpha-2b inhibiting scar formation
    Liu Zhendong, Wang Rui, Li Xiaolei, Wang Jingcheng
    2021, 25 (2):  317-321.  doi: 10.3969/j.issn.2095-4344.2984
    Abstract ( 422 )   PDF (623KB) ( 46 )   Save
    BACKGROUND: Interferon is a kind of cellular regulatory substance similar to polypeptide hormone, which has a wide range of biological activities, such as anti-virus, anti-proliferation, antitumor and immunomodulatory functions.
    OBJECTIVE: To review the effect of interferon α-2b to inhibit scar formation in multiple organs, so as to provide a theoretical basis for clinical treatment of scar and fibrosis diseases.
    METHODS: A computer-based online search of WanFang, VIP, CNKI, PubMed, Web of Science and Elsevier Science Direct databases was performed to retrieve papers published during 1950-2020 with the search terms of “IFNα-2b, interferon α-2b, fibroblasts, scar, hypertrophic scar, keloid” in Chinese and English, respectively. A total of 42 papers were included for the final analysis.
    RESULTS AND CONCLUSION: Interferon α-2b can inhibit the occurrence and development of scar, reduce the production of extracellular matrix collagen, inhibit the proliferation of fibroblasts, and act on cytokines and gene targets related to fibrosis. Findings from this study provide a theoretical basis for the clinical application of interferon α-2b in the treatment of scar formation.
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    Gingival retraction: application profiles and hot spotlights
    Li Rong, Zuo Enjun
    2021, 25 (2):  322-328.  doi: 10.3969/j.issn.2095-4344.2957
    Abstract ( 436 )   PDF (708KB) ( 151 )   Save
    BACKGROUND: The peri-implant tissue is very different from the periodontal tissue of natural teeth, so the response to the mechanical and chemical stimulation during gingival retraction must be different.
    OBJECTIVE: To review the clinical effects and application status of all kinds of gingival retraction systems, and describe the study prospect of gingival retraction around implants, attempting to help clinicians choose a better gingival retraction method.
    METHODS: Based on Chinese search terms “gingival retraction; gingival retraction time; gingival displacement effect” and “peri-implant tissue; adhesive; temporary crown; wedge-shaped defect; laser,” the authors searched and matched relevant documents in CNKI, WanFang database, VIP database from 2007 to 2019. And with the English search terms “gingival retraction; gingival; implant; temporary crown,” the authors searched for relevant literature in PubMed database. After reading title, abstract and full-text selection of literature, 60 articles were finally included for review.
    RESULTS AND CONCLUSION: Gingivlal retraction is a kind of technique that makes gingival tissue retract and produces both horizontal and vertical displacement, which separates free gingival margin from tooth surface. It has a requirement of at least 0.2 mm gingival sulcus width in order to create a clean, dry and clear-view working environment. It is convenient for clinicians to prepare the shoulder, make impression and repair wedge-shaped defect. The clinicians can also improve the edge suitability of restoration and restoration effect, reducing the complications. The gingival retraction process itself is a sort of stimulation to periodontal tissue and it causes a certain degree of damage to periodontal tissue. The damage will not recover to the pre-gingival level and cause slight permanent gingival recession. According to the amount of permanent gingival recession, clinicians can determine the position of abutment shoulder to prevent the exposure of crown edge from affecting the aesthetics. There are many factors that affect the effect of gingival retraction, the dentist needs to select the suitable gingival retraction method for the patient according to the patient’s oral and systemic conditions and in combination with the advantages and disadvantages of gingival retraction methods, in order to minimize the damage to the periodontal tissue. At present, there are many methods of gingival retraction, which can reach the required gingival sulcus width.
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