Objective Radiation-induced brain injury (RIBI) is a common complication of radiotherapy for the head and neck tumors, and the current treatment methods are limited. Repetitive transcranial magnetic stimulation (rTMS), as a non-invasive neural regulation technique, has shown great potential in neuroprotection. However, the parameter selection and biological safety of rTMS in the prevention and treatment of RIBI have not been reported. Methods Using a mouse model of RIBI, this study employed three rTMS frequencies (5, 10, and 25 Hz) for intervention. Biochemical and pathological assays were conducted to identify the optimal stimulation parameter. Subsequently, this parameter was used to evaluate the biological safety in normal mice. Results Under the conditions of this experiment, rTMS interventions with all three frequencies could reduce the levels of serum brain injury markers (NSE and S100B) and inflammatory factors in mice (P < 0.001), and alleviate the morphological and structural damage of hippocampal tissue. The 10 Hz rTMS could significantly promote hippocampal neurogenesis in RIBI mice (P < 0.05). Furthermore, 10 Hz rTMS showed no significant effects on the cognitive function and mood of normal mice. The intervention did not significantly change the morphology and structure of the main organs, blood biochemical indicators, and the level of hippocampal neurogenesis in mice. Conclusion The 10 Hz rTMS is optimal for the prevention and treatment of RIBI with high biological safety.

Objective:To investigate the effect of thoracic X-ray irradiation on the spermatogenesis of adult male mice.Methods:A total of 24 healthy adult male C57BL/6 mice (6-8 weeks old) were randomly divided into radiation group (Radiation) and sham-radiation group (Sham), 12 mice in each group. The area of thoracic irradiation was 1.5 cm× 2 cm, and the dose rate was 3.04 Gy/min, 8 Gy/d for 3 consecutive days, 24 Gy in total. At 7 d and 21 d after thoracic irradiation, the bilateral testes and epididymal tails were stripped and the testicular index was calculated. The morphology of testis was examined by haematoxylin-eosin (HE) staining, then the diameter of seminiferous tubules and the thickness of seminiferous epithelium were measured. The sperms were collected from the bilateral epididymal tails for sperm counting. The level of apoptosis in testis and levels of apoptosis-related proteins were detected by TUNEL and Western blot, respectively.Results:Compared with Sham group, the morphology of testis and epididymis was seriously damaged, the diameter of seminiferous tubules significantly decreased at 21 d after irradiation ( t = 8.93, P < 0.05), and the seminiferous epithelium significantly decreased at 7 d and 21 d after irradiation ( t = 4.24, 12.77, P < 0.05). In addition, the number of sperms significantly decreased ( t = 4.30, 2.98, P < 0.05). The number of TUNEL positive cells in the seminiferous epithelium significantly increased at 7 d and 21 d after irradiation ( t = -2.73, -3.74, P < 0.05). Meanwhile, the level of cleaved Caspase-3 protein significantly increased at 7 d and 21 d after irradiation ( t = -2.96, -2.46, P < 0.05). The concentrations of SCF and GDNF did not change at 7 d after irradiation, but were significantly increased at 21 d after irradiation ( t = -10.46, -5.42, P < 0.05). Conclusions:The thoracic X-ray irradiation could lead to spermatogenesis disorder in male adult mice, and the induction of spermatogenic cell apoptosis and the secretory dysfunction of sertoli cells may be involved.

Objective:To study the effects and mechanism of hyperbaric oxygen (HBO) on apoptosis and growth factor of skeletal muscle in rats with type 2 diabetes.Methods:A total of 24 GK rats with blood sugar >16.7 mmol/L, selected from 32 GK rats after 1 week feeding, were divided into model group ( n=8), metformin group ( n=8), and HBO group ( n=8), according to random number table method. The left 8 rats with blood sugar ≤ 16.7 mmol/L were included into control group. The rats in the control group and the model group were irrigated with pure water 5 ml·kg -1·d -1. The rats in the HBO group were given HBO every day on the basis of the treatment of the control group and the model group. After 3 weeks, the rats were executed. The tail blood was collected to measure fasting blood glucose. The right gastrocnemius muscle tissues were collected and made into paraffin sections for HE staining. The morphological and structural changes of gastrocnemius muscle in each group were observed under a light microscope. Immunohistochemistry was used to detect the expressions of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), endothelial nitric oxide synthase (eNOS), cytokine, and Caspase-3; and the integrated optical density (IOD) was measured and calculated for comparison. TUNEL method was used to detect gastrocnemius cell apoptosis, and the apoptosis index was calculated. Results:After the rats were executed at 21 d, the 12 h fasting blood glucose of the rats in the model group, the metformin group, and the HBO group were all significantly higher than that of the control group [(7.26±0.78, 6.90±0.74, 6.92±0.95 vs. 4.74±0.53) mmol/L, P<0.05 in each comparison]. The expression of skeletal muscle bFGF in the rats of the control group was the highest, while that of the model group was the lowest. The expressions of skeletal muscle bFGF in the rats of the metformin group and the HBO group were obviously higher than that of the model group, but lower than that of the control group, with statistically significant difference ( P<0.05). The expressions of skeletal muscle VEGF in the rats of the metformin group and the HBO group were obviously higher than those of the model group and the control group, with statistically significant difference ( P<0.05). The expression of skeletal muscle eNOS in the rats of the control group was the highest, while those of the model group, the HBO group, and the metformin group were low. There were no statistical differences among the four groups ( P>0.05). The expressions of skeletal muscle Caspase-3 and the gastrocnemius cell apoptosis indexes of the metformin group and the HBO group were significantly lower than those of the model group, but higher than those of the control group, with statistical difference ( P<0.05) in all comparison. Conclusion:HBO can improve the skeletal muscle structure of the rats with diabetes, improve the expressions of bFGF and VEGF, decrease the expression of Caspase-3, and reduce cell apoptosis; it, therefore, has a certain protective effect on the skeletal muscle in rats with diabetes.

Objective:To study the effects and mechanism of hyperbaric oxygen (HBO) on apoptosis and growth factor of skeletal muscle in rats with type 2 diabetes.Methods:A total of 24 GK rats with blood sugar >16.7 mmol/L, selected from 32 GK rats after 1 week feeding, were divided into model group ( n=8), metformin group ( n=8), and HBO group ( n=8), according to random number table method. The left 8 rats with blood sugar ≤ 16.7 mmol/L were included into control group. The rats in the control group and the model group were irrigated with pure water 5 ml·kg -1·d -1. The rats in the HBO group were given HBO every day on the basis of the treatment of the control group and the model group. After 3 weeks, the rats were executed. The tail blood was collected to measure fasting blood glucose. The right gastrocnemius muscle tissues were collected and made into paraffin sections for HE staining. The morphological and structural changes of gastrocnemius muscle in each group were observed under a light microscope. Immunohistochemistry was used to detect the expressions of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), endothelial nitric oxide synthase (eNOS), cytokine, and Caspase-3; and the integrated optical density (IOD) was measured and calculated for comparison. TUNEL method was used to detect gastrocnemius cell apoptosis, and the apoptosis index was calculated. Results:After the rats were executed at 21 d, the 12 h fasting blood glucose of the rats in the model group, the metformin group, and the HBO group were all significantly higher than that of the control group [(7.26±0.78, 6.90±0.74, 6.92±0.95 vs. 4.74±0.53) mmol/L, P<0.05 in each comparison]. The expression of skeletal muscle bFGF in the rats of the control group was the highest, while that of the model group was the lowest. The expressions of skeletal muscle bFGF in the rats of the metformin group and the HBO group were obviously higher than that of the model group, but lower than that of the control group, with statistically significant difference ( P<0.05). The expressions of skeletal muscle VEGF in the rats of the metformin group and the HBO group were obviously higher than those of the model group and the control group, with statistically significant difference ( P<0.05). The expression of skeletal muscle eNOS in the rats of the control group was the highest, while those of the model group, the HBO group, and the metformin group were low. There were no statistical differences among the four groups ( P>0.05). The expressions of skeletal muscle Caspase-3 and the gastrocnemius cell apoptosis indexes of the metformin group and the HBO group were significantly lower than those of the model group, but higher than those of the control group, with statistical difference ( P<0.05) in all comparison. Conclusion:HBO can improve the skeletal muscle structure of the rats with diabetes, improve the expressions of bFGF and VEGF, decrease the expression of Caspase-3, and reduce cell apoptosis; it, therefore, has a certain protective effect on the skeletal muscle in rats with diabetes.

Objective: To explore the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection who received entecavir alone or in combination with anluohuaxianwan for 78 weeks. Methods: Patients with chronic HBV infection were randomly treated with entecavir alone or in combination with anluohuaxian for 78 weeks. Ishak fibrosis score was used for blind interpretation of liver biopsy specimens. The improvement in liver fibrosis condition before and after the treatment was compared. Student's t test and non-parametric test (Mann-Whitney U-Test and Kruskal-Wallis test) were used to analyze the measurement data. The categorical variables were analyzed by Chi-square test method and Spearman’s rank correlation coefficient was used to test bivariate associations. Results: Liver fibrosis improvement rate after 78 weeks of treatment was 36.53% (80/219) and the progression rate was 23.29% (51/219). The improvement of liver fibrosis was associated to the degree of baseline fibrosis and treatment methods (P < 0.05). The improvement rate of hepatic fibrosis in patients treated with anluohuaxianwan combined with entecavir at baseline F < 3 (54.74%, 52/95) was significantly higher than that in patients treated only with entecavir (33.33%, 16/48), P = 0.016 and the progression rate of hepatic fibrosis (13.68%, 13/95) was lower than that in patients treated alone (18.75%, 9/48), P = 0.466. In patients with baseline F < 3, the proportion of patients with improved and stable liver fibrosis in the combined treatment group (68.1%, 32/47) was higher than that in the treatment group alone (51.7%, 15/29). Conclusion Combined anluohuaxianwan and entecavir treatment can significantly improve the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection. Furthermore, it has the tendency to improve the stability rate and reduce the rate of progression of liver fibrosis.

Objective: To study the correlation between the level of T-bet expression and liver damage in peripheral plasma cells of patients with autoimmune hepatitis (AIH) in order to provide reference for the study of pathogenesis and development of diseases. Methods: The peripheral venous blood and clinical examination data of 29 cases with AIH and 6 healthy volunteers were collected. The percentage of subpopulations of peripheral blood B cells and the proportion of T-bet⁺ cells in each subgroup were detected by flow cytometry. Plasma cells (CD19⁺CD10^-CD27^hiCD38^hi), primary B cells (CD19⁺CD10^-CD27^-IgD⁺), transitional B cells (CD19⁺CD10⁺), and memory B cells (CD19⁺CD10^-CD27⁺IgD^-) were the included subsets of B cells. Serum immunoglobulin G (IgG) and alanine aminotransferase (ALT) levels, the proportion of B cells in peripheral blood subsets and IgG level, the proportion of T-bet⁺ cells in each subset and the proportion of T-bet⁺ plasma cells in each subset in B cells, the proportion of T-bet⁺ plasma cells and the level of serum ALT were analyzed for correlation analysis. Statistical analysis was performed using two independent sample t-tests and linear regression. Results: The serum IgG level of AIH patients with abnormal ALT (19.47 ± 1.039)g/L was significantly higher than that of normal ALT patients (15.5 ± 1.069)g/L, and the difference was statistically significant (t = 2.65, P < 0.05). The percentage of peripheral plasma cells in B cells of AIH patients (2.80 ± 0.14) % was higher than that of healthy volunteers (0.73 ± 0.09) %, and the difference was statistically significant (P < 0.01). The percentage of T-bet⁺ cells in peripheral plasma cells of AIH patients (23.54 ± 1.61) % was higher than that of healthy volunteers (6.59±0.59) % , and the difference was statistically significant (P < 0.01). The correlation analysis showed that the proportion of T-bet⁺ cells in peripheral plasma cells of AIH patients was positively correlated with the proportion of plasma cells to B cells (r = 0.224 7, P < 0.01), and the percentage of peripheral plasma cells to B cells was positively correlated with the level of serum IgG (r = 0.299 1, P < 0.01). Serum IgG level was correlated with the level of ALT, reflecting an indicator of liver damage (t = 2.65, P < 0.05). Conclusion The increase of T-bet expression in the peripheral plasma cells of AIH patients is associated with liver damage, which is a new mechanism of AIH pathogenesis and disease progression.

BACKGROUND:At present, the incidence rates of knee joint diseases such as knee osteoarthritis, knee joint degenerative are high. The major clinical treatment is total knee replacement in the clinic, so it is necessary to evaluate the changes in stress and bone mineral density of the regions surrounding the prosthesis after replacement. ＜br> OBJECTIVE:To explore periprosthetic stress and bone mineral density and to analyze their correlation after total knee arthroplasty. ＜br> METHODS:A total of 20 cases undergoing total knee arthroplasty were chosen.The hospital for special surgery scores were used to evaluate patients’ functional recovery at 12 months after total knee arthroplasty. The periprosthetic femur was divided into four regions of interest (ROI), respectively ROI 1-4;periprosthetic tibia was divided into three regions of interest, respectively ROI 5-7. Stress surrounding the prosthesis was analyzed using three-dimensional finite element analysis at 1, 3, 6 months, 1, 2, 3 years after replacement. Simultaneously, bone mineral density surrounding the prosthesis was measured using dual-energy X-ray absorptiometry. ＜br> RESULTS AND CONCLUSION:No patients affected infection or loosening of the prosthesis. At 12 months after replacement, the score of hospital for special surgery was (90.23±2.37), which showed significant differences as compared with before replacement (39.68±1.31) (P＜0.05). The level of stress shielding was highest in ROI 5 and lowest in ROI 3. Stress shielding rate of ROI increased with statistical difference at 6 months after operation (P＜0.05). At 1, 2, 3 years after operation, shielding rate in periprosthetic femoral stress in ROI 1 decreased. Compared with 1 month after operation, the difference was statistical y significant (P＜0.05). However, shielding rate of tibial periprosthetic stress in ROI 6 increased. Compared with 1 month after operation, the difference was statistical y significant (P＜0.05). Bone mineral density after 1 month after operation had no significant decrease (P>0.05). At 3 months after operation, bone mineral density began to decline significantly (P＜0.01). The decrease was most obviously in ROI 5 and the change was least in ROI 3. After 1 year of operation, bone mineral density did not change significantly. These data indicated that changes in bone mineral density were correlated with stress shielding after total knee arthroplasty. Monitoring two variations can provide theoretical data for preventing bone loss, which provides references for clinical rehabilitation guidance.