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18 November 2022, Volume 26 Issue 32 Previous Issue    Next Issue

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Construction of a nomogram model for predicting bone mass loss in middle-aged men Shen Lianwei, Wang Wei 2022, 26 (32):  5085-5090.  doi: 10.12307/2022.891 Abstract ( 317 )   PDF (886KB) ( 41 )   Save BACKGROUND: A nomogram prediction model can be plotted based on independent influencing factors derived from multiple regression analyses. The probability of bone mass loss in patients can be intuitively calculated based on the total score obtained from various influencing factors. OBJECTIVE: To establish a nomogram prediction model based on the independent factors of bone mass loss in middle-aged men.  METHODS: The clinical data of 279 middle-aged men who underwent physical examinations in the Physical Examination Center of the First Affiliated Hospital of Jinzhou Medical University from June 15 to July 15, 2021 and from October 15 to November 15, 2021 were collected and divided into training set (214 cases) and verification set (65 cases) according to different time periods of data collection. The self-made “Middle-aged Male Bone Mass Survey Questionnaire” was used in all subjects and the DiscoveryW dual-energy X-ray bone densitometer was used to measure the bone mineral density of the left hip. Based on the single-factor analysis and multivariate Logistics regression analysis of training set, the independent influencing factors of bone mass loss were screened, and the nomogram model for predicting bone mass loss was made. C-index verification and calibration curve were used to preliminarily evaluate the differentiation and calibration degree of the model. The internal validation of the model was performed by k-fold cross validation, and the external validation of the model was performed by the validation set. The performance of the nomogram model was further evaluated by C index, Hosmer-Lemeshow test, and calibration curve. RESULTS AND CONCLUSION: Results from the single-factor analysis showed that there were statistical differences in smoking, dairy products, exercise, and breakfast in the training set. Multivariate Logistics regression analysis results showed that smoking was an independent risk factor for bone mass loss, while dairy products and exercise were protective factors. According to the independent factors and existing theories, the nomogram model for predicting bone mass loss was established with a C-index of 0.671 (95% confidence interval: 0.600-0.750), indicating that the calibration curve fitted well. K-fold cross verification showed that the mean value of C-index was 0.708. The C-index of external validation was 0.689 (95% confidence interval: 0.600-0.742), P=0.09 > 0.05 in the Hosmer-Lemeshow test, and the calibration curve was fitted well. To conclude, this nomogram model for predicting bone mass loss has well inspection efficiency and can help screen middle-aged male patients with boss mass loss. Figures and Tables | References | Related Articles | Metrics

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Shikonin intervenes with osteoclast formation and osteoclast-related gene expression Bo Yujia, Lin Jing, Wang Liping, Zhao Jin 2022, 26 (32):  5126-5131.  doi: 10.12307/2022.837 Abstract ( 531 )   PDF (1082KB) ( 37 )   Save BACKGROUND: The activation of osteoclast plays an important role in the occurrence and development of periodontitis. In recent years, shikonin has been used as an adjuvant drug after periodontal non-surgical treatment because of its small side effects and anti-inflammatory properties.    OBJECTIVE: To discuss the effects of shikonin on osteoclast formation induced by receptor activator for nuclear factor-κB ligand (RANKL) and osteoclast-related gene expression. METHODS: The toxicity of different concentrations of shikonin (0, 0.062 5, 0.125, 0.25, 0.5, 1, and 2 μmol/L) to RAW264.7 cells was detected by cell counting kit-8 assay, and the optimal concentration was determined. RAW264.7 cells were induced into osteoclasts by different concentrations of RANKL (0, 30, 50, 100 μg/L). The number of osteoclasts was revealed by tartrate resistant acid phosphatase staining, and the optimal concentration of RANKL was determined. After treatment with different concentrations of shikonin, the effects of RANKL on osteoclast formation were detected by tartrate resistant acid phosphatase staining and tartrate resistant acid phosphatase activity. The mRNA expressions of osteoclast marker genes, including matrix metalloproteinase 9, cathepsin K, receptor activator for nuclear factor-κB, nuclear factor of activated T cells and proto-oncogene c-Fos, were detected by real-time quantitative PCR.  RESULTS AND CONCLUSION: Shikonin at a concentration of higher than 0.5 μmol/L significantly inhibited the growth of RAW264.7 cells (P < 0.05). The optimal induction concentration of RANKL was 50 μg/L. Shikonin inhibited osteoclast formation in a concentration-dependent manner (P < 0.05). Shikonin also inhibited the expression of matrix metalloproteinase 9, cathepsin K, receptor activator for nuclear factor-κB, nuclear factor of activated T cells, and proto-oncogene c-Fos in a concentration-dependent manner (P < 0.05). To conclude, shikonin can inhibit RANKL-induced osteoclast formation by inhibiting the expression of osteoclast-related genes in a concentration-dependent manner in vitro. Figures and Tables | References | Related Articles | Metrics

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Emodin promotes fracture healing in osteoporotic fracture rats by inhibiting miR-338-3p expression Li Haoliang, Li Dongfang 2022, 26 (32):  5155-5161.  doi: 10.12307/2022.879 Abstract ( 290 )   PDF (1384KB) ( 65 )   Save BACKGROUND: Osteoporosis is characterized by low bone mass and osseous degenerative changes. These conditions increase the risk of fractures. Emodin has potent anti-inflammatory effects and can affect bone metabolism, but its specific mechanism of action is unknown. OBJECTIVE: To investigate the effects and related mechanisms of emodin on the healing of osteoporotic fracture rats.  METHODS: Fifty-three female Sprague-Dawley rats were randomly divided into sham (n=5) and osteoporotic (n=48) groups. An osteoporotic model was created by ovariectomy and then the right femur of each model rat was crosscut. The model rats in the osteoporotic group were further randomized into six groups: simple fracture group, low-, middle-, and high-dose emodin groups, high-dose emodin+NC mimic group, and high-dose emodin+miR-338-3p mimic group, with 8 rats in each group. Emodin was intragastrically administered at a dose of 10, 20, and 40 mg/kg, once daily for 8 weeks; NC mimic and miR-338-3p mimic were injected into the callus at a dose of 2 mg/kg, once a day, for 7 continuous days. After 8 weeks of intragastrical administration, the bone mineral density of the rat femur at the midshaft and lumbar vertebrae was analyzed. The levels of serum inflammatory factors including NO, tumor necrosis factor α, and interleukin 1β were detected using ELISA. Histomorphological examination was performed using hematoxylin-eosin staining. The level of miR-338-3p in the rat callus was determined with RT-qPCR. The effects of different doses of emodin on osteogenesis-related genes and proteins involved in the ERK/BMP/Smad5 signaling pathway in the rat callus were determined by western blot analysis.  RESULTS AND CONCLUSION: Compared with the simple fracture group, emodin increased the bone mineral density of the femoral midshaft and lumbar vertebrae in a dose-dependent manner (P < 0.05). The bone mineral density of the femoral midshaft and lumbar vertebrae in the high-dose emodin+miR-338-3p mimic group was significantly higher than that in the simple fracture group but lower than that in the high-dose emodin group (P < 0.05). Hematoxylin-eosin staining results showed that compared with the simple fracture group, emodin increased the trabecular bone area in the femoral tissue in a dose-dependent manner (P < 0.05). The trabecular bone area in the high-dose emodin+miR-338-3p mimic group was larger than that in the simple fracture group but lower than that in the high-dose emodin group. Compared with the simple fracture group, emodin increased serum nitric oxygen level and decreased serum tumor necrosis factor α and interleukin 1β levels in a dose-dependent manner (P < 0.05). The levels of serum inflammatory factors in the high-dose emodin+miR-338-3p mimic group were higher than those in the simple high-dose emodin group but lower than those in the fracture group. Compared with the simple fracture group, emodin decreased the expression of miR-338-3p mRNA in the callus in a dose-dependent manner (P < 0.05). Compared with the simple fracture group, emodin dose-dependently increased the protein expression of alkaline phosphatase, osteocalcin, Runx2, bone morphogenetic protein 2, and phosphorylated Smad5 (P < 0.05) and reduced the protein expression of phosphorylated ERK1/2 and phosphorylated JNK in the callus. To conclude, emodin could dose-dependently promote fracture healing in osteoporotic fracture rats, probably by suppressing the expression of miR-338-3p and activating the ERK/JNK/BMP/Smad signaling pathway.  Figures and Tables | References | Related Articles | Metrics

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Anti-osteoporosis effect of Gushukang in ovariectomized rats: a lumbar metabonomic analysis Zhang Haiyong, Huang Jingwen, Xie Bingying, Chen Sainan, Xie Lihua, Chen Xuan, Li Shengqiang, Ge Jirong 2022, 26 (32):  5185-5190.  doi: 10.12307/2022.937 Abstract ( 799 )   PDF (1349KB) ( 33 )   Save BACKGROUND: The research on the specific molecular mechanisms and therapeutic targets of Gushukang on osteoporosis is insufficient, which needs further research and discussion. OBJECTIVE: To analyze and identify the changes of lumbar metabolites in ovariectomized rats, explore the therapeutic effect of Gushukang on osteoporosis in ovariectomized rats and discuss its potential mechanism by using the non-targeted metabonomics of ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass, thereby to further explore the specific mechanisms and targets of Gushukang in the treatment of osteoporosis. METHODS: Female Sprague-Dawley rats, which were ovariectomized to establish osteoporosis model, were divided into model group, Gushukang group, and sham operation group. Osteoporotic models were established using ovariectomy. Four weeks after modeling, rats in the Gushukang group were given Gushukang by gavage; and those in the sham operation and model groups were given the equal volume of normal saline. After 12 weeks of continuous gavage, the lumbar metabolites of rats were analyzed by the ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass technology, and the common differential metabolites were selected for bioinformatics analysis using Kyoto Encyclopedia of Genes and Genomes pathway.  RESULTS AND CONCLUSION: A total of 24 common significant differential metabolites were identified in the three groups. Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that Gushukang could regulate important pathways such as fatty acid metabolism, glycerol phospholipid metabolism and amino acid metabolism, and participate in the protection of bone metabolism in osteoporosis model rats. These findings indicate that Gushukang exerts its anti-osteoporosis effect by interfering with lipid metabolism and amino acid metabolism, so as to achieve the purpose of preventing osteoporosis. Figures and Tables | References | Related Articles | Metrics

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In vitro isolation, culture and identification of chondrocytes from the ankle joint of aged mice Jia Caixia, He Si, Ha Xiaoqin 2022, 26 (32):  5097-5101.  doi: 10.12307/2022.909 Abstract ( 350 )   PDF (992KB) ( 27 )   Save BACKGROUND: Knee cartilage loss certainly occurs in aged mice, so it is difficult to extract the articular cartilage from the mice. In particular, it is difficult and rarely reported to extract chondrocytes from the small joint on the articular cartilage surface of the ankle joint.   OBJECTIVE: To explore the effect of an improved two-step enzyme digestion method to extract chondrocytes from the ankle joint of aged mice, and to observe and evaluate their biological characteristics in vitro.  METHODS: The ankle joints of 46-week-old mice were selected, separated and extracted sequentially by the improved two-step enzymatic digestion method in the laboratory with 0.25% pancreatin EDTA and 2g/L type II collagenase. Extracted cells were cultured in vitro and cell growth morphology was observed with an inverted microscope. After trypan blue staining, the number of primary adherent cells was calculated. Toluidine blue and type II collagen immunofluorescence staining was conducted. Chromogenic method using dimethyl methylene blue was used to determine the content of glycosaminoglycan sulfate in the primary adherent cell culture and digestive fluid. Proliferation of chondrocytes was detected using cell counting kit-8 to identify and evaluate the biological characteristics of chondrocytes. RESULTS AND CONCLUSION: After the two-step enzymatic digestion of the articular cartilage, all cells were dissociated and released. An average of 9×105 primary adherent cells were harvested from each ankle joint of aged mice. Primary cells and the first- and second-generation cells displayed spindle and polygonal morphology. Toluidine blue staining showed blue-purple particles that were easily dyed. Type II collagen immunofluorescence staining revealed the positive expression of red fluorescence. The contents of glycosaminoglycan sulfate in the primary adherent cell culture fluid and digestive fluid were (42.41±15.00) and (65.63±10.00) mg/L, which were (0.35±3.78) and (0.21±2.56) mg/L in the negative control culture fluid and digestive fluid, respectively. The number of primary cells reached the maximum on the 6th day, and the number of the first- and second-generation cells reached the maximum on the 5th day. Meanwhile, the cell viability of the first-generation cells was greater than that of the second-generation cells and the primary cells. To conclude, the improved two-step enzymatic digestion method can successfully isolate chondrocytes from the ankle joint of aged mice. Large amounts of adherent cells with high purity and good biological characteristics can successfully synthesize and secrete type II collagen and proteoglycan. The first-generation chondrocytes have the highest viability, followed by the second-generation cells, which can be used for experimental research. Figures and Tables | References | Related Articles | Metrics

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Activation of chondrocyte degeneration related signaling pathways and expression of differential factors in ATDC5 mouse cells after silent information regulator 1 knockdown Xie Xiaochen, Yang Qi, Weng Jian, Zeng Hui, Kang Bin, Liu Pei, Zhong Huage, Yu Fei 2022, 26 (32):  5112-5118.  doi: 10.12307/2022.849 Abstract ( 480 )   PDF (1249KB) ( 50 )   Save BACKGROUND: Chondrocytes degeneration often leads to common orthopedic diseases such as osteoarthritis, intervertebral disc degeneration and meniscus degeneration. SIRT1 gene is closely related to chondrocytes degeneration. When SIRT1 gene is knocked down in ATDC5 mouse chondrocytes, it can affect cell function via signaling pathways related to chondrocyte degeneration, and then affect the process of orthopedic diseases. OBJECTIVE: To investigate the significantly activated signaling pathways related to chondrocyte degeneration and the expression of differential factors in ATDC5 mouse chondrocytes after SIRT1 gene knockdown. METHODS: The lentivirus carrying SIRT1 gene or not was transfected into ATDC cells. The transfected cells were divided into control (transfected with negative control lentivirus) and experiment group (transfected with lentivirus carrying SIRT1 gene). Total RNA was extracted for quality testing. Gene chip was used to detect the expression of coding RNA in the cells. Combined with bioinformatics analysis, we identified significantly activated signaling pathways related to chondrocyte degeneration, and elaborated differentially expressed factors in some signaling pathways. RESULTS AND CONCLUSION: Forty-two signaling pathways were significantly activated in ATDC5 cells after SIRT1 gene knockdown. Among them, we selected five signaling pathways related to chondrocyte degeneration and reported less, including hepatocyte growth factor signaling pathway (ETS1, PIK3CA, NRAS, PIK3C2A, FGFR2, ELF1, FGFR3, CDKN1A, AKT3, FRS2, PRKD3, MAP3K2; Ratio=0.139 0, Z-score=2.673), neuregulin signaling pathway  (RPS6KB1, STAT5A, MTOR, BTC, HBEGF, RNF41; Ratio=0.136 0, Z-score=2.309), insulin receptor signaling pathway (TSC1, EIF4EBP1, PRKAR2B, PPP1R12A, TSC2; Ratio=0.106 0, Z-score=2.138), chemokine receptor 4 signaling pathway (PAK4, EGR1, RHOJ, RHOB, PLCB1, GNG5; Ratio=0.097 0, Z-score=2.500), actin cytoskeleton signaling pathway (ABI2, BRK1, PIP4K2B, MYLK, IQGAP1, ARPC1A, ACTA2, EZR, SSH2, ACTG1, IQGAP3; Ratio=0.092 1, Z-score=2.236), so as to provide novel targets for treating chondrocyte degeneration related diseases. Figures and Tables | References | Related Articles | Metrics

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Effect of inhibiting miR-203-3p expression on bone formation around dental implants in diabetic mice Li Kongmei, Luo Yicai, Li Hao 2022, 26 (32):  5102-5106.  doi: 10.12307/2022.906 Abstract ( 295 )   PDF (1184KB) ( 31 )   Save BACKGROUND: Up to now, dental implant has become an important restoration for patients with tooth loss, and bone formation around dental implants is the key to the success of dental implantation. There is often a poor bone formation around dental implants under diabetic conditions and the mechanism remains unclear. Recent studies have shown that miR-203-3p regulates bone formation in different diseases, and strongly expressed in jaw bone tissues under diabetic conditions. However, it is unclear whether miR-203-3p can influence bone formation around dental implants. OBJECTIVE: To investigate the effect of inhibiting miR-203-3p expression on bone formation around dental implants in diabetic mice. METHODS: Twelve mice with normal blood glucose were randomly selected as normal group from forty-eight C57BL/6J mice. The other 36 mice were selected to make diabetic models and then randomly divided into diabetic group, diabetes+antago miR-NC group, and diabetes+antago miR-203-3p group (n=12 per group). Single implant was inserted into the left maxilla of each mouse. Diabetic+antago miR-NC group was injected with antago miR-203-3p negative control, antago miR-NC (0.25 mmol/L). Diabetes+antago miR-203-3p group was injected with antago miR-203-3p (0.25 mmol/L). The other two diabetic groups were injected with the equal volume of normal saline. After 3 months, the left maxillary bone tissue of mice was collected, and the bone formation around dental implants was detected using micro-CT and hematoxylin-eosin staining. qRT-PCR was used to detect the gene expression of miR-203-3p, Smad1, alkaline phosphatase, Runx2, Osterix, and type I collagen in bone tissue around dental implants.   RESULTS AND CONCLUSION: Micro-CT results showed that compared with the diabetic group and diabetes+antago miR-NC group, the diabetes+antago miR-203-3p group showed better alveolar bone formation around dental implants and higher bone volume fraction and bone trabecular thickness (P < 0.05). Hematoxylin-eosin staining results showed that the Lane-Sandhu score of bone formation around dental implants was higher in the diabetes+antago miR-203-3p group than the diabetic group and the diabetes+antago miR-NC group (P < 0.05). qRT-PCR results showed that the expression level of miR-203-3p gene in bone tissue around dental implants was lower (P < 0.05), while the expression levels of Smad1, alkaline phosphatase, Runx2, Osterix, and type I collagen genes were higher in the diabetes+antago miR-203-3p group than the diabetic group and the diabetes+antago miR-NC group (P < 0.05). All these findings indicate that the inhibition of miR-203-3p expression is beneficial to bone formation around dental implants in diabetic hyperglycemia, and these effects may be related to the regulation of Smad signaling pathway and the expression of osteogenesis-related factors such as alkaline phosphatase, Runx2, Osterix, and type I collagen. Figures and Tables | References | Related Articles | Metrics

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Characteristics of the growth, development and microarchitecture of condyle subchondral bone in rats Liu Chun, Jia Ying, Yang Shirong, Ding Qi, Yang Hua, Chen Bo 2022, 26 (32):  5162-5166.  doi: 10.12307/2022.1027 Abstract ( 387 )   PDF (1038KB) ( 64 )   Save BACKGROUND: The growth and development of the condyle is an important factor affecting the health of the temporomandibular joint. The condyle subchondral bone, as the final presentation area of condylar bone reconstruction and bone remodeling, has a more active basis for remodeling. However, previous studies have not fully explored the subchondral bone of the condyle and its microarchitecture. OBJECTIVE: To study the microarchitecture, growth and development characteristics of rat condyle subchondral bone by Micro-CT.  METHODS: Twenty-four Sprague-Dawley rats of both sexes, aged 3 weeks, were randomly divided into 0-month group, 3-month group, 5-month group, and 7-month group (6 rats per group). After execution under anesthesia, the right mandibular condyle of each rat was cut out for Micro-CT scans, in which the condyle was divided into nine grids, with the middle anterior and posterior subchondral bone as the regions of interest to detect the bone microstructure parameters. RESULTS AND CONCLUSION: Most parameters of female rats changed significantly at the age from 0 to 7 months (P < 0.05). From 3 to 5 months of age, there were significant changes in bone mineral density, bone surface area to bone volume ratio, trabecular number, connectivity, and changes density (P < 0.05), and some parameters began to level off. By 7 months, only bone volume fraction and total porosity still showed significant differences (P < 0.05). For male rats aged from 0 to 7 months, only trabecular pattern factor, trabecular separation, structural model index, and connectivity changed insignificantly with time (P > 0.05). From 3 to 5 months of age, most parameters of male rats changed steadily (P > 0.05). For male rats at the age of 7 months, bone mineral density, trabecular number, connectivity, and trabecular thickness changed significantly (P < 0.05). From 3 to 7 months of age, there were significant changes in bone morphology, trabecular morphology and spatial structure of trabecular bone in male and female rats, but these changes in males were greater than those in females. The above results explain that the condyle subchondral bone is dense and dominated by the lamellar bone; and the condyle subchondral bone develops earlier and presents with a steady growth earlier in female rats than male rats.  Figures and Tables | References | Related Articles | Metrics

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Application of digital smile design based on case-based reasoning in aesthetic restoration of anterior teeth Zhu Guohui, Mu Ya 2022, 26 (32):  5191-5195.  doi: 10.12307/2022.896 Abstract ( 386 )   PDF (833KB) ( 62 )   Save BACKGROUND: To achieve a unified aesthetic restoration effect, digital smile design technologies have been introduced and gradually applied in clinical practice. However, due to the limitations and authenticity of the software’s own dental pictures, the promotion of the software encountered difficulties. Researchers found that using the digital smile design software to import previous similar cases may optimize this problem, making the design of the prosthesis authentic and personalized. OBJECTIVE: To verify the advantages and limitations of the case-based digital smile design method by comparing the effect of the case-based digital smile design method and the traditional aesthetic restoration method in the aesthetic restoration of anterior teeth. METHODS: From October 2018 to October 2020, 36 patients requiring aesthetic restoration of anterior teeth at the Department of Prosthodontics in Tianjin Stomatological Hospital were selected and randomly divided into experimental group and control group according to the sequence of treatment, with 18 patients in each group. In the experimental group, digital smile design technology was used to collect and import digital photos, and the diagnostic wax pattern and diagnostic decoration were made based on the pictures of the prostheses from the most similar cases in the past. Patients could obtain the digital smile design drawing and three-dimensional oral preview form for doctor-patient communication and design adjustment. The control group was treated with digital photography combined with colorimetric palettes. Immediately after aesthetic repair, patient satisfaction evaluation was performed. Aesthetic prostheses were clinically evaluated as per the modified United States Public Health Service (USPHS) criteria at 2 weeks, 6 months, and 12 months after aesthetic repair.  RESULTS AND CONCLUSION: In terms of patient satisfaction, patients in the experimental group were more satisfied with the shape and color of the prosthetics than those in the control group (P < 0.05), and there was no significant difference in the comfort of the prosthetics and the time to seek medical treatment (P > 0.05). There was no significant difference between the two groups in postoperative prosthesis integrity, periodontal healthy, tooth sensitivity, edge fit, color matching, and occlusal relationship at 2 weeks after restoration (P > 0.05), while the experimental group was superior to the control group in color matching at 6 and 12 months after restoration (P < 0.05). To conclude, the case-based digital smile design technique is superior to the traditional method in doctor-patient communication, prosthesis color and shape; however, there are limitations such as the limited number of previous cases and the increased workload and cost of the digital smile design process.  Figures and Tables | References | Related Articles | Metrics

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Functional differentiation and visualization of nerve fibers in the human sciatic nerve Li Xiaojun, Li Jia, Gao Feng, Li Lang, Li Qin, Ouyang Xiangyu 2022, 26 (32):  5148-5154.  doi: 10.12307/2022.925 Abstract ( 398 )   PDF (1208KB) ( 95 )   Save BACKGROUND:  It is the precondition of nerve recovery that nerve tracts are accurately stitched after sciatic nerve injury. However, it is unclear about the trajectory of the distal and proximal fracture ends of nerve tracts in different functional regions, and thus it is unable to accurately fix and repair the fracture end, which is the difficulty of surgical treatment, leading to fault regeneration of different functional nerve tracts. It has a great influence on postoperative neurological recovery.  OBJECTIVE: To study a set of methods for the reconstruction of motor and sensory nerve fibers in peripheral nerves, so as to provide a feasible method for seeking the distribution and trajectory of motion fibers and other fibers in the sciatic nerve and to provide the basis for accurate suture after nerve injury. METHODS: In patients with upper-middle thigh amputation, a 1 cm sample of the sciatic nerve was taken 20 cm from the upper femoral condyle, marked using hair, fixed with paraffin, and then sliced continuously. Following immunohistochemistry and immunofluorescence staining, the nerve sections were scanned under a microscope to obtain two-dimensional image information. The muscular branch of the sural nerve and the cutaneous branch of the superficial peroneal nerve were taken from the same limb as negative and positive controls. Using CINEMA 4D software developed by Maxon Computer (Germany), the stained sections were used to reconstruct the three-dimensional images of the internal movement of the sciatic nerve and other nerve fibers. RESULTS AND CONCLUSION: The specific antibody of motor fibers in the sciatic nerve, Anti-HB9/HLXB9 Antibody, was used to stain motor fibers, while other fibers were not dyed. Using CINEMA 4D software, two-dimensional images could be used to form three-dimensional images of nerve fibers in the sciatic nerve, and then motor fibers could be accurately distinguished from the other fibers using immunofluorescent staining. It is suggested that motor nerve fiber bundles and other nerve fiber bundles in the sciatic nerve can be accurately distinguished after immunohistochemical staining. To conclude, advanced three-dimensional reconstruction software can be used to accurately reconstruct the internal structure of the sciatic nerve, to provide an effective method for accurately differentiating motor fibers and other fibers and exploring the trajectory of nerve functional bundles. Figures and Tables | References | Related Articles | Metrics

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Skin administration of Wentong Huoxue Cream can relieve inflammation in diabetic peripheral neuropathy rats Ye Ting, Li Jing, Xu Lijuan, Ma Li 2022, 26 (32):  5091-5096.  doi: 10.12307/2022.893 Abstract ( 480 )   PDF (1079KB) ( 85 )   Save BACKGROUND: Wentong Huoxue Cream can improve oxidative stress, reduce blood viscosity, restore microcirculation, relieve diabetic peripheral nerve injury, and inhibit sensory conduction and pain formation, but its regulatory mechanism remains unclear. OBJECTIVE: To observe the effects of Wentong Huoxue Cream on JNK signaling pathway proteins and inflammatory factors in diabetic peripheral neuropathy rats.  METHODS: A rat model of diabetic peripheral neuropathy was established in adult Wistar rats. Forty diabetic peripheral neuropathy rats were randomly divided into five groups (n=8 per group): model group, low-, middle- and high-dose Wentong Huoxue Cream groups (given 2 mL of 0.75, 1.5, 3.0 g/mL Wentong Huoxue Cream to the lower limbs of rats, respectively) and Western medicine group (given 2 mL of control cream). Skin treatment in each group was performed twice a day for 28 days. Additionally, six rats were taken as normal group. After the last administration, nerve conduction tests were performed and the expressions of interleukin-6, interleukin-18, tumor necrosis factor α, high-sensitivity C-reactive protein, and myelin basic protein in serum were detected by ELISA. The expression of JNK and P-JNK protein in sciatic ganglia was detected by western blot and the expression of JNK mRNA was determined by RT-PCR.  RESULTS AND CONCLUSION: The velocity of sensory nerve conduction and motor nerve conduction was significantly decreased in the model group compared with the normal group (P < 0.05). Compared with the model group, the sensory nerve conduction velocity increased significantly after intervention with different doses of Wentong Huoxue Cream (P < 0.05), especially in the high-dose Wentong Huoxue Cream group (P < 0.05). Compared with the model group, the motor nerve conduction velocity was significantly increased in the high-dose Wentong Huoxue Cream group and Western medicine group (P < 0.05). Compared with the normal group, the serum levels of interleukin-6, tumor necrosis factor α, high-sensitivity C-reactive protein, and myelin basic protein were significantly increased (P < 0.05) and the serum level of interleukin-18 was significantly decreased in the model group (P < 0.05). Compared with the model group, the serum levels of interleukin-6, interleukin-18, tumor necrosis factor α, high-sensitivity C-reactive protein, and myelin basic protein were significantly decreased in all the treatment groups (P < 0.05), especially in the high-dose Wentong Huoxue Cream group and Western medicine group. Compared with the normal group, p-JNK protein and JNK mRNA expression levels were significantly increased in the model group (P < 0.05). Compared with the model group, p-JNK protein and JNK mRNA expression levels were significantly decreased in the middle- and high-dose Wentong Huoxue Cream groups and Western medicine group (P < 0.05). Therefore, Wentong Huoxue Cream can inhibit the abnormal activation of JNK signal transduction pathway, reduce inflammatory reactions, and protect the peripheral nerve function of diabetic peripheral neuropathy rats. Figures and Tables | References | Related Articles | Metrics

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Effect of endothelial Piezo1 knockout on steroid-induced osteonecrosis of the femoral head in mice Pan Zhaofeng, Yang Junzheng, He Qi, Zhang Gangyu, Xiao Jiacong, Chen Baihao, Wang Haibin, Chen Peng 2022, 26 (32):  5107-5111.  doi: 10.12307/2022.1026 Abstract ( 519 )   PDF (1213KB) ( 56 )   Save BACKGROUND: One of the important causes of steroid-induced osteonecrosis of the femoral head is the damage of vascular endothelial cells caused by glucocorticoids, which subsequently leads to ischemia and hypoxia of the femoral head. Piezo1 plays an important role in promoting angiogenesis and homeostasis. However, the effect of endothelial Piezo1 on steroid-induced osteonecrosis of the femoral head is unclear. OBJECTIVE: To observe the effects of endothelial Piezo1 on steroid-induced osteonecrosis of the femoral head. METHODS: Then male Piezo1fl/fl mice, 8 weeks of age, were randomly divided into two groups: control group and model group. Five male Piezo1ΔEC mice were selected in an experimental group, in which lipopolysaccharide at a dose of 20 μg/kg was intraperitoneally injected for 2 continuous days, followed by intramuscular injection of methylprednisolone sodium succinate 40 mg/kg for 4 weeks. The control group was injected with the same amount of PBS at the same site. Mice were sacrificed at 12 weeks of age and bone samples of both lower limbs were obtained. Micro-CT and hematoxylin-eosin staining were used to observe and analyze the changes of bone microstructure and histomorphological changes of the femoral head in each group, and then SPSS software was used for statistical analysis. RESULTS AND CONCLUSION: Compared with the model group, the damage of bone microstructure was significantly aggravated in the experimental group. Pathological findings further showed that the number of empty bone lacunae was significantly increased in the experimental group compared with the model group. Several bone morphometric parameters like trabecular bone volume and trabecular number were significantly reduced (P < 0.001) and trabecular separation increased significantly in the experimental group compared with the model group (P < 0.000 1). To conclude, endothelial Piezo1 can participate in the process of steroid-induced osteonecrosis of the femoral head in mice and endothelial Piezo1 gene deletion can aggravate steroid-induced osteonecrosis of the femoral head in mice. Figures and Tables | References | Related Articles | Metrics

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Characteristic changes of intestinal mucosal morphology and transcriptomics in silent information regulator 6 gene knockout mice You Yihui, Song Chunhui, Chen Xi, Yao Xiaohong, Ke Junyu, Du Qun, Song Ning, Li Yanwu 2022, 26 (32):  5119-5125.  doi: 10.12307/2022.847 Abstract ( 395 )   PDF (2076KB) ( 140 )   Save BACKGROUND: Intestinal mucosal renewal is a key physiological process to maintain body homeostasis, and stem cells at the crypt base are the driving force for intestinal epithelial proliferation and differentiation. Silent information regulator (SIRT) is involved in the regulation of cell gene repair, metabolism, energy balance, and lifespan. Research on its role in intestinal epithelial homeostasis has gradually become a hot spot. Intestinal organoids are formed by 3D culture of single crypt stem cells, which can present the intestinal epithelial renewal and repair process after damage in vitro. It is the latest model for studying intestinal mucosal renewal.  OBJECTIVE: To elucidate the characteristic changes of intestinal mucosal morphology, intestinal organoid formation and its transcriptomics in a SIRT6 gene-specific knockout (SIRT6-i/-i) mouse model.  METHODS: The Cre-loxp method was used to establish the SIRT6-i/-i mouse mode. Paraffin sections of small intestine tissue were prepared to observe the pathological changes using hematoxylin-eosin staining, and the expression of intestinal stem cell marker Lgr5 and Paneth cell marker Lyso was detected by immunohistochemistry. The intestinal organoid SIRT6-i/-i mouse model was constructed in vitro, SIRT6 protein expression in intestinal organoids was detected by western blot assay, and the protein expression of Lgr5, cytokeratin 20 and mucin 2 in intestinal organoids was detected by immunofluorescence staining. The intestinal tissue was used for transcriptome sequencing, and the characteristic changes of the transcriptome in the mouse model were analyzed. RESULTS AND CONCLUSION: Compared with the control group, SIRT6-i/-i mouse intestinal mucosa showed sparse and shortened villi, shallower crypts, increased number of goblet cells, significantly decreased number of Paneth cells, and significantly reduced expression of Lgr5 and Lyso. The expression of SIRT6 protein in the SIRT6-i/-i mouse organoid was significantly reduced. The budding rate of organoids and the expression of Lgr5 were reduced, while the expression of absorption epithelial cell marker cytokeratin 20 and goblet cell marker mucin 2 increased obviously. Transcriptome results showed that there were 846 differentially expressed genes between the control and model mouse intestine, including 438 up-regulated genes and 408 down-regulated genes. The top five differentially expressed genes enriched in the Kyoto Encyclopedia of Genes and Genomes pathway were chemical carcinogens, drug metabolism, retinoic acid metabolism, linoleic acid metabolism, and steroid biosynthesis. And the differentially expressed genes in the top five pathways focusing on the Cyp450 family included Cyp2c29, Cyp2c65, Cyp3a11, and Cyp3a25. To conclude, SIRT6 gene deletion can lead to the imbalance of intestinal stem cell proliferation and differentiation, and the mechanism may be related to the metabolic regulation pathways of SIRT6. Regulating cytochrome P450 family by SIRT6 is closely related to the homeostasis of intestinal mucosal renewal. Figures and Tables | References | Related Articles | Metrics

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Network pharmacology and proteomics analysis of Jiawei Xiaoyao San in the treatment of liver cancer complicated with depression in rats Wen Xiaoyu, Sun Yuhao, Li Zhuoxian, Xu Lijing, Xia Meng 2022, 26 (32):  5132-5142.  doi: 10.12307/2022.905 Abstract ( 399 )   PDF (1870KB) ( 288 )   Save BACKGROUND: Previous research has shown that Jiawei Xiaoyao San can regulate the content of dopamine and 5-hydroxytryptamine in the hippocampus and significantly improve the pathological morphology of the liver and depression symptoms in rats with liver cancer complicated with depression. However, its pharmacological mechanism has not been fully elucidated. OBJECTIVE: Through network pharmacological analysis and proteomic testing, to screen out the effective targets of Jiawei Xiaoyao San for treating liver cancer complicated with depression and to discuss the active components and possible mechanism of Jiawei Xiaoyao San in the treatment of liver cancer complicated with depression.  METHODS: Sprague-Dawley rat models of liver cancer complicated with depression were established and treated with Jiawei Xiaoyao San. Proteomics and network pharmacology analyses were performed to identify the targets for differentially expressed proteins in serum. Gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were conducted for effective targets. Liver morphology was detected using hematoxylin-eosin staining. Depression-like behavioral changes were observed in the rat models. Enzyme-linked immunosorbent assay was used to determine the serum level of related targets.  RESULTS AND CONCLUSION: Three possible common protein targets were identified based on proteomic and network pharmacology analyses: glutathione transferase (GSTM1), 3-phosphoinositide dependent protein kinase 1 (PDK1), heat shock protein 90AB1 (HSP90AB1). Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses found that these targets were directly involved in the negative regulation of neuronal apoptosis, PI3K-Akt signaling pathway, and prostate cancer molecular signaling pathway. Hematoxylin-eosin staining and behavioral tests showed that Jiawei Xiaoyao San improved liver damage and depression-like behaviors in rats. Results from the enzyme-linked immunosorbent assay revealed that GSTM1, PDK1, and HSP90AB1 had corrective effects after intervention with Jiawei Xiaoyao San. To conclude, Jiawei Xiaoyao San may correct depression-like behavior and improve liver morphology by regulating the expression of GSTM1, PDK1, and HSP90AB1 in rats with liver cancer complicated with depression. Figures and Tables | References | Related Articles | Metrics

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Establishing a mouse model of allergic rhinoconjunctivitis Gong Yubo, Shi Yuanyuan, Li Yuanchao, Lu Wenjun, Xia Liping, Guo Xiaohua, Luo Ling, Wu Wei 2022, 26 (32):  5143-5147.  doi: 10.12307/2022.890 Abstract ( 406 )   PDF (1055KB) ( 125 )   Save BACKGROUND: Animal models for studying the pathogenesis of allergic conjunctivitis and allergic rhinitis have been well developed. Allergic rhinitis and allergic conjunctivitis often coexist in clinical practice, that is, allergic rhinoconjunctivitis. However, the animal model for studying its pathogenesis is rarely reported. OBJECTIVE: To establish a mouse model of allergic rhinoconjunctivitis for exploring its pathogenesis. METHODS: Twenty female C57 mice (SPF grade) were randomly divided into two groups: control group and experimental group. On days 0, 4, 7, 14, and 21, each mouse in the experimental group was intraperitoneally given 0.2 mL of injectable suspension containing ovalbumin and adjuvant Al(OH)3. After an interval of 3 days, the eyes and nose of each mouse were given 5% ovalbumin, 10 μL per eye or per nose, 5 continuous days per week to induce allergic symptoms. Mice in the control group were given the same amount of PBS. Ocular and nasal symptoms were observed and scored in the sensitization and provocation stages. The serum levels of ovalbumin-specific IgE, total IgE, and histamine were detected by ELISA method. Pathological changes of the palpebral conjunctiva and nasal mucosa in mice were observed using hematoxylin-eosin staining. RESULTS AND CONCLUSION: Allergic symptoms of the nose and eyes were induced in all the mice in the experimental group. The time from provocation until  the emergence of allergic symptoms of the nose and eyes was (10.500±1.080) days and (20.300±2.058) days respectively. The levels of ovalbumin-specific IgE, total IgE, and histamine in the experimental group were significantly higher than those in the control group (P < 0.05). There were remarkable pathological changes in the palpebral conjunctiva and nasal mucosa in the experimental group. To conclude, the mouse model of allergic rhinoconjunctivitis is successfully established.  Figures and Tables | References | Related Articles | Metrics

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Microtubule-associated protein 1 light chain 3 is involved in the occurrence and development of steroid-induced avascular necrosis of the femoral head in a rabbit model Ma Hailong, Zhao Zhenqun, Liu Wanlin, Sun Jun 2022, 26 (32):  5167-5172.  doi: 10.12307/2022.930 Abstract ( 308 )   PDF (960KB) ( 71 )   Save BACKGROUND: Steroid-induced avascular necrosis of the femoral head is still a difficult problem, and it is of great significance to study its pathogenesis. OBJECTIVE: To discuss the role of autophagy gene microtubule-associated protein 1 light chain 3 (MAP1LC3) in steroid-induced avascular necrosis of the femoral head and the effect of 3-methyladenine on MAP1LC3. METHODS: A total of 36 healthy New Zealand white rabbits, aged 5 months, were randomly divided into 3 groups, with 12 rabbits in each group. Control group was given intramuscular injection of normal saline, 2 mL per rabbit, once a week, four times in total. Model group was given intramuscular injection of 4 mg/kg methylprednisolone, once a week, four times in total. 3-Methyladenine group was given intramuscular injection of 4 mg/kg methylprednisolone followed by intramuscular injection of 3-methyladenine, 2 μL per rabbit, once a week, four times in total. Three animals from each group were sacrificed at 1, 2, 3, and 4 weeks for sampling. The femoral heads of the bilateral hindlimbs were taken. Empty bone lacunae were counted under optical microscope. mRNA and protein expression levels of MAP1LC3 were detected by RT-PCR and western blot assay, respectively.  RESULTS AND CONCLUSION: Compared with the control group, the rate of empty lacunae increased significantly in the model and 3-methyladenine groups (P < 0.05), but the rate of empty lacunae was significantly lower in the 3-methyladenine group than the model group (P < 0.05). The expression of MAP1LC3 mRNA in the model group was higher than that in the  3-methyladenine group (P < 0.05), indicating the activation of autophagy is inhibited by 3-methyladenine. These findings indicate that MAP1LC3 is expressed in the animal model, confirming that autophagy plays a certain role in the pathogenesis of steroid-induced avascular necrosis of the femoral head; 3-methyladenine inhibits the expression of MAP1LC3 and reduces the occurrence and development of steroid-induced avascular necrosis of the femoral head. Figures and Tables | References | Related Articles | Metrics

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Bone metabolism in a rat model of diabetes mellitus intervened by Xianling Gubao Capsules Jiao Yinghua, Bao Kairan, Song Jieqiong, Xing Lei, Cui Lihua, Gao Jingyuan, Qi Ning, Liu Xiangyu 2022, 26 (32):  5173-5178.  doi: 10.12307/2022.941 Abstract ( 538 )   PDF (1258KB) ( 115 )   Save BACKGROUND: Xianling Gubao Capsules (a commonly used Chinese medicine in orthopedic surgery) can improve bone metabolism and prevent osteoporosis; however, whether it has a preventive effect or a possible mechanism on osteoporosis complicated by diabetes has been rarely reported. OBJECTIVE: To investigate the effects of Xianling Gubao Capsules on bone metabolism and related osteogenic marker factors Wnt3a, β-catenin and bone morphogenetic protein-2 in rats with diabetes mellitus complicated by osteoporosis and to explore their mechanisms of action. METHODS: Thirty-six male healthy 8-week-old Sprague-Dawley rats were treated with streptozotocin at a dose of 60 mg/kg to establish a diabetes mellitus model and were then randomly divided into model group, Xianling Gubao Capsule group and alendronate group, with 12 rats in each group. Another 12 normal rats were used as the normal group. Rats in the Xianling Gubao Capsule group and alendronate group were given Xianling Gubao Capsule and alendronate sodium suspension by gavage for 12 weeks, respectively. Rats in the normal and model groups were gavaged with the same amount of double-distilled water for 12 weeks. Blood glucose level of model rats was measured once a week during the administration period to ensure the accuracy of the experiment. Femur, tibia and systemic bone mineral density in each group were measured by dual energy X-ray absorptiometry. Hematoxylin-eosin staining was used to observe the morphological changes of the right femur in each group. Immunohistochemistry and RT-PCR were applied to detect the expression of osteogenic marker factors Wnt3a, β-catenin, and bone morphogenetic protein-2 at protein and mRNA levels.   RESULTS AND CONCLUSION: Compared with the model group, the femur, tibia and systemic bone mineral density values of rats increased significantly in the Xianling Gubao Capsule group (P < 0.05). The mRNA and protein expressions of Wnt3a, β-catenin, and bone morphogenetic protein-2 in bone tissues decreased significantly in the model group (P < 0.05). While compared with the model group, the mRNA and protein expressions of Wnt3a, β-catenin, and bone morphogenetic protein-2 were significantly increased in the Xianling Gubao Capsule group (P < 0.05). In addition, there was no significant difference in the expressions of above-mentioned indicators between the Xianling Gubao Capsule group and alendronate group. These findings confirmed that Xianling Gubao Capsules exert a protective effect on bone metabolism in diabetic rats by the Wnt /β-catenin signaling pathway. Figures and Tables | References | Related Articles | Metrics

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Construction of bromodomain-containing protein 4 gene knockout mice by CRISPR/Cas9 combined with Cre-loxP technology Wang Xiangyu, Zhu Ruizhi, Zhao Zhiping, Zhang Yuda, Zhang Yongtao, Wang Changyao 2022, 26 (32):  5179-5184.  doi: 10.12307/2022.892 Abstract ( 789 )   PDF (1321KB) ( 86 )   Save BACKGROUND: Currently, the combined use of CRISPR/Cas9 and CRE-LOXP to prepare bromodomain-containing protein 4 (BRD4) gene knockout mice is very rare.  OBJECTIVE: To construct a BRD4 gene knockout mouse model by using CRISPR/Cas9 technology combined with Cre-loxP technology that can be used to knock out the BRD4 gene fragment in the mouse genome.  METHODS: According to the exon sequence of BRD4 gene, a gRNA was designed and synthesized. After gRNA was transcribed in vitro, Cas9 mRNA and plasmid containing loxp site were injected into fertilized oocytes. Cas9 cut the target fragment by recognizing the leading strand of the gRNA, and then loxp was inserted into the cleavage site. After breeding with Cre, the Cre enzyme cut the loxp site to achieve the final specific deletion effect. The fertilized oocytes were transplanted into C57BL/6N recipient female mice to obtain progeny mice. The offspring mice were sequenced to identify their genotypes. The BRD4-loxP +/- (F0 generation) mice with loxp loci being successfully introduced was bred with wild-type C57BL/6N mice, and the stably inherited mice BRD4-loxP +/- (F1 generation) were selected. Some of the BRD4-loxP +/- mice interbred with each other and some interbred with CAGGcre-ERTM mice. BRD4-loxP +/+ mice were co-expressed with BRD4-loxP+/- and CAGGcre-ERTM mice (F2 generation). The F2 generation mice were bred to obtain homozygous BRD4-loxP +/+ and CAGGcre-ERTM mice. BRD4 gene knockout was induced by intraperitoneal injection of tamoxifen (75 mg/kg, dissolved in corn oil) in 6-8 weeks homozygous mice for 7 consecutive days. DNA was extracted from the tail fragment of the knockout mice using adsorption column method. The expression of BRD4 gene fragment in mouse tail tissue was detected by agarose gel electrophoresis. RESULTS AND CONCLUSION: Screening using PCR revealed that mice 7, 8, 9, 10, 12, 14, 16, 19, 20, 21, 25, 42, 43, and 47 of F1 generation were identified as heterozygous mice that were successfully inserted with two alleles of loxP fragment. The F3 homozygotes bred from F2 generation were induced with tamoxifen, and PCR gel electrophoresis verified that the BRD4 gene fragment was successfully knocked out in the homozygous mice. We therefore successfully constructed BRD4 gene knockout mice using CRISPR/Cas9 technology combined with CRE-LOXP technology in this study.  Figures and Tables | References | Related Articles | Metrics

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Hypothalamus: the key controller for exercise-improved bone metabolism Lu Pengcheng, Liu Bo, Jin Shengjie, Tian Zhikai, Zeng Xinyu, Chen Xianghe 2022, 26 (32):  5201-5208.  doi: 10.12307/2022.884 Abstract ( 424 )   PDF (1188KB) ( 108 )   Save BACKGROUND: Bone metabolism has always been a hotspot in bone tissue engineering research. With the deepening of the mechanism, it has been found that hypothalamus is the main regulatory center for exercise to improve bone metabolism.  OBJECTIVE: To review the role of hypothalamus in bone metabolism and to explore the mechanism of exercise-mediated hypothalamus to improve bone metabolism. METHODS: A computer-based search of CNKI and PubMed databases was performed for relevant articles published from January 1983 to December 2021 with “exercise; hypothalamus; bone metabolism” as keywords both in Chinese and English. The documents that did not meet the inclusion criteria were excluded, and the 72 selected documents were summarized. RESULTS AND CONCLUSION: The hypothalamus is the key to regulating the body’s physiological activity and the downstream of the hypothalamus-pituitary-target gland axis plays an important regulatory roleto influence hormone secretion, such as neuropeptide Y, leptin, osteocalcin and calcitonin, thus regulating regulatory factors and signaling pathways for bone metabolism, and further improving common bone diseases, such as osteoporosis. In recent years, the bone-building effect mediated by “brain-bone” crosstalk has received widespread attention in medicine, biology and sports science, but its specific occurrence and mechanism of action are still under investigation. Exercise activates bone metabolic signaling pathways through mechanical force stimulation and conduction of intracellular electrical signals. Figures and Tables | References | Related Articles | Metrics

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Signal peptide-CUB-epidermal growth factor-like domain protein family as coreceptors/ligands is involved in embryonic development and disease development Wang Zijie, Tian Zhihui, Wu Jiayuan 2022, 26 (32):  5209-5216.  doi: 10.12307/2022.916 Abstract ( 346 )   PDF (1035KB) ( 118 )   Save BACKGROUND: Signal peptide-CUB-EGF-like domain-containing protein (SCUBE) can act as a coreceptor and/or a ligand to activate signaling pathways, such as bone morphogenetic protein, vascular endothelial growth factor, transforming growth factor β, and hedgehog, and plays an important role in a variety of physiological and pathological processes OBJECTIVE: To review relevant literature worldwide and summarize the research progress in the structure and function of SCUBE family, as well as their roles in embryonic development and disease occurrence. METHODS: The first author used a computer to search for related articles included in PubMed and CNKI databases from 2000 to 2021. The search terms were “SCUBE, EGF family, structure and function, coreceptor, signaling pathways, embryonic development, disease, mechanism, tumor, protein, gene” in English and Chinese, respectively. According to the inclusion criteria, 68 articles were selected for review. RESULTS AND CONCLUSION: The SCUBE family includes three independent members, SCUBE1, SCUBE2, and SCUBE3. SCUBE family members share five similar domains, including one amino-terminal signal peptide sequence, nine tandemly arranged epidermal growth factor-like repeats, one amino glycosylation spacer, three cysteine-rich domains, and one carboxy-terminal CUB-like domain. SCUBE family members can act as coreceptors and/or ligands, binding through their special CUB-like domains to activate multiple signaling pathways including bone morphogenetic protein, transforming growth factor β and vascular endothelial growth factor and produce corresponding biological effects. SCUBE family members are widely expressed in a variety of tissues and organs, and have a dynamic expression pattern during vertebrate embryonic development. The SCUBE family is closely related to the occurrence of diseases, and can be used as key therapeutic targets for diseases such as bone and tooth dysplasia, vascular dysplasia, autoimmune diseases and tumors. SCUBE family members play an important role in embryonic development and disease occurrence. They can not only be used as therapeutic targets for diseases, but also have broad application prospects in the field of tissue engineering regeneration in the future. However, their specific functions and mechanisms still need to be further studied.  Figures and Tables | References | Related Articles | Metrics

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Autophagy and inflammation-induced alveolar bone metabolism Song Na, Liu Guanjuan, Luo Shanshan, Huo Hua, Cheng Yuting, Hong Wei, Liao Jian 2022, 26 (32):  5217-5222.  doi: 10.12307/2022.917 Abstract ( 358 )   PDF (871KB) ( 179 )   Save BACKGROUND: Autophagy can remove damaged proteins or excessive organelles, help cells adapt to adverse environmental changes, and maintain intracellular environment stability and self-renewal. However, abnormalities or dysfunctions in autophagy will lead to bone metabolic diseases. OBJECTIVE: To review the effect of autophagy on inflammation-induced alveolar bone metabolism. METHODS: The first author searched PubMed, Web of Science, Geenmedical, WanFang, and CNKI databases for relevant articles published from January 2011 to November 2021. The search terms were “autophagy, osteoblasts, osteoclasts, NLRP3, bone metabolism” in English and Chinese, respectively. Finally, 63 articles were included in result analysis after screening, analyzing and summarizing. RESULTS AND CONCLUSION: Autophagy has a protective effect on the body. Moderate activation of autophagy is conducive to maintaining the internal environment homeostasis in bone tissue, and disorder or abnormality of autophagy pathways may disturb the dynamic balance during bone remodeling, resulting in the occurrence of alveolar bone metabolic diseases. NLRP3 inflammasomes and autophagy have a bidirectional regulatory effect and autophagy may play an important role in the occurrence and development of metabolic diseases in the alveolar bone by regulating NLRP3 inflammasomes. Figures and Tables | References | Related Articles | Metrics

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Relationship between iron metabolism and ischemic stroke Fan Yongfu, Su Kaiqi, Yuan Jie, Nie Chenchen, Ruan Xiaodi, Duan Zhaoyuan, Feng Xiaodong 2022, 26 (32):  5223-5228.  doi: 10.12307/2022.918 Abstract ( 305 )   PDF (1151KB) ( 119 )   Save BACKGROUND: Ischemic stroke is a major clinically fatal and disabling disease and there is no effective solution yet. Abnormal iron metabolism is related to various physiological and pathophysiological processes, and the latest discovery is that it is closely related to ischemic stroke. OBJECTIVE: To explore the relationship between iron metabolism and ischemic stroke and look for potential new therapeutic targets for ischemic stroke from a novel perspective, in order to guide the treatment of ischemic stroke. METHODS: We searched for relevant literature from January 2010 to January 2022 in PubMed, CNKI, and WanFang databases. The search terms were “iron metabolism, ischemic stroke, reactive oxygen species, glutathione peroxidase 4, blood-brain barrier, inflammatory factors, excitatory amino acid, Tau, oligodendrocytes, autophagy, iron death, serum ferritin” in English and Chinese. Finally, 82 articles were included for further analysis.  RESULTS AND CONCLUSION: (1) The balance of iron metabolism is essential to maintain the normal life activities of brain neurons. Iron metabolism imbalance can participate in a variety of pathophysiological processes to further aggravate secondary brain injury after cerebral ischemia. (2) Treatments of ischemic stroke with iron metabolism as a target can effectively reduce secondary brain injury after cerebral ischemia and promote nerve function recovery. (3) At present, there are relatively few studies on the relationship between secondary brain injury and iron after ischemic stroke. The detailed pathophysiological mechanism has not yet been elucidated, and related treatment strategies need to be further improved. Therefore, it is necessary to further study the mechanism of iron metabolism after ischemic stroke, which may provide a new target for the treatment of secondary brain injury after ischemic stroke. Figures and Tables | References | Related Articles | Metrics

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Growth and development characteristics of dental and basal arch in children with skeletal class III malocclusion Gao Liping, Tang Li, Wan Lu, Liu Yan, Li Xiaobing, Zhong Wenyi 2022, 26 (32):  5229-5235.  doi: 10.12307/2022.897 Abstract ( 536 )   PDF (976KB) ( 125 )   Save BACKGROUND: At present, the diagnosis and treatment of skeletal class III malocclusion are challenging, mainly because of the diversity and unpredictability of craniofacial structure growth in such patients. Treatment decisions and their success or failure much depend on the future growth potential of patients with skeletal class III malocclusion.  OBJECTIVE: To review the research progress in growth and development of dental and basal arch in children with skeletal class III malocclusion. METHODS: Literature retrieval was conducted in PubMed, CNKI, WanFang, and Medline using the keywords of “malocclusion; skeletal class III malocclusion; dental arch; basal arch; growth and development; mixed dentition; tongue; etiology; jaw” in English and Chinese, and studies and experiments of low quality and irrelevant to the topic were excluded. Finally, 64 articles were included for careful reading and analysis. RESULTS AND CONCLUSION: Skeletal class III malocclusion is a common malocclusion type in children with abnormal occlusal development, and the jaw, dental arch, and basal arch are also affected by genetic factors to a certain extent. As special craniomaxillofacial complexes, the jawbone, dental arch and basal arch have the same growth trend and there was a synergistic effect between them. In children with skeletal class III malocclusion, the growth rate of the maxilla and mandible is related to age and bone age, and the growth rate of the jaw bone length varies at different growth peaks until adulthood. It is particularly important to correctly predict the peak growth period and inhibit the growth and development of the mandible in children with skeletal class III malocclusion. Due to the complexity of its treatment, timely detection of abnormal development of the upper and lower jaws, teeth, dentition and occlusal relationship, correct diagnosis, early correction, and timely intervention are of great significance to guide the normal growth and development of the craniomaxillofacial region. However, the current research on the growth and development of the dental arch and basal bone arch in children mainly focuses on the normal occlusion, because the growth and development characteristics of children with skeletal class III malocclusion cannot be observed once treatment is started. Therefore, further studies are required on the growth and development mechanism of skeletal class III malocclusion.  Figures and Tables | References | Related Articles | Metrics

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Periosteum distraction for the treatment of diabetic foot ulcer: theoretical basis and clinical verification Liu Jie, Hua Qikai, Li Shanlang, Yu Jie, Su Hongjie, Ding Yi, Zhao Yongxin, Su Yongfeng, Chen Yan 2022, 26 (32):  5236-5241.  doi: 10.12307/2022.910 Abstract ( 897 )   PDF (1471KB) ( 92 )   Save BACKGROUND: Periosteal distraction has been found to stimulate osteogenesis and angiogenesis; however, whether this technique can be used for the treatment of diabetic foot remains unclear.  OBJECTIVE: To review the research progress of periosteal distraction in the promotion of osteogenesis and neovascularization, to clarify the theoretical basis of this technique for the treatment of diabetic foot, and to introduce its preliminary application in the treatment of diabetic foot.  METHODS: Literature retrieval was conducted using computer in WanFang, CNKI, PubMed and Elsevier databases using the keywords of “distraction histogenesis, distraction osteogenesis, tibia transverse transport and diabetic foot, periosteal distraction osteogenesis, periosteal distraction and diabetic foot” in Chinese and English. Finally, 42 articles were included for analysis.  RESULTS AND CONCLUSION: Based on Ilizarov’s law of tension-stress or the principle of distraction histogenesis, distraction osteogenesis has been used to treat bone defects, nonunion and other orthopedic diseases. The angiogenesis accompanying distraction osteogenesis gave birth to the technique of tibial transverse transfer, which has been successfully applied to refractory lower extremity ulcers including diabetic foot. Also based on the principle of distraction histogenesis, studies have found that a continuous, stable and slow distraction of the periosteum can promote bone and vascular regeneration, that is, the phenomenon of periosteal distraction osteogenesis and angiogenesis. The mechanism of this phenomenon may be related to the structure and composition of the periosteum. However, the current research on periosteal distraction mainly focuses on its osteogenic function in animal experiments but hardly in humans. Like tibial transverse transfer to promote angiogenesis for the treatment of diabetic foot, periosteal distraction angiogenesis may also be used to treat diabetic foot, which is the theoretical basis of periosteal distraction for the treatment of diabetic foot. Our preliminary clinical study has shown that this technique is effective in the treatment of diabetic foot. Periosteal distraction is simpler and shorter than the lateral tibial bone transfer because it does not require osteotomy. The periosteal distraction technique lacks the manipulation of tibial drilling and bone transfer and lacks the “fenestration decompression” of the bone marrow cavity. So, the periosteal distraction technique may be inferior to the lateral bone transfer of the tibia in terms of promoting angiogenesis and wound healing. In the future, the indications, contraindications, and specific mechanism of periosteal distraction still need to be explored. Figures and Tables | Related Articles | Metrics

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Long non-coding RNA and osteoporosis Kang Teng, Lan Fengjun, Qin Hao, Liu Gang 2022, 26 (32):  5242-5247.  doi: 10.12307/2022.921 Abstract ( 342 )   PDF (978KB) ( 79 )   Save BACKGROUND: In recent years, it has been found that the expression of long non-coding RNA molecules is closely related to the occurrence and development of osteoporosis and plays an important role in it. However, the underlying molecular mechanism has not yet been clarified. OBJECTIVE: To review the role and mechanism of long non-coding RNA in the occurrence and development of osteoporosis, in an attempt to provide a new idea and approach for long non-coding RNA in the prevention and treatment of osteoporosis METHODS: PubMed, CNKI, and WanFang databases were searched for relevant literature using the keywords of “long non-coding RNA, osteoporosis, mesenchymal stem cells, osteoblasts, osteoclasts, cellular pathways” in English and Chinese. Fifty-nine articles were enrolled for further analysis after excluding obsolete, repetitive and unreliable literature. RESULTS AND CONCLUSION: Long non-coding RNAs can be involved in the prevention and treatment of osteoporosis by regulating and inhibiting the osteogenic differentiation of bone marrow and adipose mesenchymal stem cells. Long non-coding RNAs can promote osteoblasts and inhibit the proliferation of osteoclasts to influence the occurrence and development of osteoporosis. The regulation of long non-coding RNAs induces the activation or inhibition of osteoblast-related cellular pathways, thus altering the expression of target genes to participate in the procession of osteoporosis. Therefore, further studies on the role of long non-coding RNA in the pathogenesis of osteoporosis are helpful to provide a new idea and method for the early prevention and clinical treatment of osteoporosis. Figures and Tables | References | Related Articles | Metrics

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A Meta-analysis of repetitive transcranial magnetic therapy in patients with post-traumatic stress disorder Song Ge, Liu Gang, Jiang Liang, Han Jiming, Zhang Guiqing 2022, 26 (32):  5196-5200.  doi: 10.12307/2022.935 Abstract ( 399 )   PDF (1110KB) ( 77 )   Save OBJECTIVE: Repetitive transcranial magnetic stimulation can effectively relieve the psychological symptoms of patients with stroke, spinal cord injury, and chronic pain. This paper systematically evaluates the therapeutic effect of repetitive transcranial magnetic stimulation in the treatment of post-traumatic stress disorder (PTSD). METHODS: Self-controlled and randomized controlled trials were searched in Embase, MEDLINE, Cochrane Library, PubMed, Web of Science, WanFang, VIP, and CNKI databases for the therapeutic effect of repetitive transcranial magnetic stimulation on PTSD patients. The search period was from the inception date to December 2021. Two researchers trained in meta-analysis were responsible for evaluating the quality of screening articles and extracting relevant data. Meta-analysis was performed on the extracted data by RevMan software. The Clinician-Administered PTSD Scale score was used to measure the severity of symptoms and injury, and the PTSD Checklist was used to evaluate the improvement of PTSD symptoms. RESULTS: Eight documents involving 339 PTSD patients were included. Meta-analysis results showed that compared with conventional treatments, repetitive transcranial magnetic stimulation significantly improved the severity and symptoms of PTSD [standardized mean difference (SMD)=-5.90, 95% confidence interval (CI) (-5.76, -4.41), P < 0.001; mean difference (MD)=-6.52, 95% CI (-9.74, -3.30), P < 0.001]. The severity and symptoms of PTSD patients were significantly improved after intervention compared with the baseline data [MD=18.22, 95% CI (10.84, 25.60), P < 0.001; MD=17.08, 95% CI (16.30, 17.86), P < 0.001]. High-frequency repetitive transcranial magnetic stimulation had no significant advantage over low-frequency repetitive transcranial magnetic stimulation in improving the severity of PTSD [MD=3.72, 95%CI (-4.55, 11.99), P=0.38], but had an advantage in the relief of PTSD symptoms [MD=6.15, 95% CI (0.15, 12.15), P=0.04].  CONCLUSION: Repetitive transcranial magnetic stimulation can effectively improve the severity and symptoms of PTSD patients. Figures and Tables | References | Related Articles | Metrics