BACKGROUND:Nerve growth factor is a protein that induces nerve growth and regulates biological behaviors such as proliferation and differentiation of mesenchymal stem cells. OBJECTIVE:To investigate the promoting effect of nerve growth factor on chondrogenic differentiation of bone marrow mesenchymal stem cells. METHODS:Rabbit bone marrow mesenchymal stem cells were isolated and cultured,and nerve growth factor was transfected into bone marrow mesenchymal stem cells by lentiviral transfection.The effects of nerve growth factor on the proliferation,migration,hypertrophic differentiation,and chondrogenic differentiation of bone marrow mesenchymal stem cells were detected by CCK-8 assay,cell scratch assay,alizarin red staining,and western blot assay,using the transfected null-loaded virus as control.To further investigate the promoting effect of nerve growth factor on the chondrogenic differentiation of bone marrow mesenchymal stem cells,interleukin 1β was added in bone marrow mesenchymal stem cells transfected with empty virus and nerve growth factor for 14 days.The expression of proteins related to chondrogenic differentiation and hypertrophic differentiation was detected by western blot assay. RESULTS AND CONCLUSION:(1)CCK-8 assay results showed that nerve growth factor had no significant effect on the proliferation of bone marrow mesenchymal stem cells.(2)Compared with the control group,overexpression of nerve growth factor enhanced the migration ability of the cells,and the expression of cartilage-associated proteins type II collagen and SOX9 was up-regulated(P<0.05),while the expression of hypertrophic-associated proteins type X collagen and Runx2 was down-regulated(P<0.05).(3)Compared with the empty virus+interleukin 1β group,the expression of cartilage-associated proteins type II collagen and Sox9 was up-regulated(P<0.05),and the expression of hypertrophy-associated proteins type X collagen and Runx2 was down-regulated after overexpression of nerve growth factor(P<0.05).(4)The results indicated that nerve growth factor could promote the chondrogenic differentiation of bone marrow mesenchymal stem cells.

Objective To explore the therapeutic effect of Dangui Quyu Pill combined with acupuncture on adenomyosis(ADS)and its influences on serum levels of vascular endothelial growth factor(VEGF)and carbohydrate antigen 199 and 125(CA199,CA125).Methods A total of 120 ADS patients were randomly divided into the control group(treated with acupuncture)and the treatment group(treated with Dangui Quyu Pills combined with acupuncture),with 60 cases in each group.The degree of dysmenorrhea before treatment and 6 months after treatment was evaluated by Visual Analogue Scale(VAS).Traditional Chinese Medicine(TCM)syndrome score was conducted.Uterine volume was detected by color Doppler ultrasound on the day of enrollment and 6 months after treatment(avoiding the menstrual period).Levels of VEGF,CA199 and CA125 were detected by ELISA before treatment and 6 months after treatment.The adverse reactions and recurrence of patients were observed and recorded.Results The total effective rate was higher in the treatment group than that of the control group(P＜0.05).After treatment,the VAS score,TCM syndrome score,menstrual volume,uterine volume,serum levels of VEGF,CA199 and CA125 were lower in both groups than those before treatment,and the treatment group was even lower than those of the control group(P＜0.05).There was no statistically significant difference in the total incidence of adverse reactions between the two groups,and the recurrence rate of the treatment group was lower than that of the control group(P＜0.05).Conclusion The combination of Dangui Quyu pill and acupuncture has obvious effect and high safety in the treatment of ADS.

Objective To explore the therapeutic effect of Dangui Quyu Pill combined with acupuncture on adenomyosis(ADS)and its influences on serum levels of vascular endothelial growth factor(VEGF)and carbohydrate antigen 199 and 125(CA199,CA125).Methods A total of 120 ADS patients were randomly divided into the control group(treated with acupuncture)and the treatment group(treated with Dangui Quyu Pills combined with acupuncture),with 60 cases in each group.The degree of dysmenorrhea before treatment and 6 months after treatment was evaluated by Visual Analogue Scale(VAS).Traditional Chinese Medicine(TCM)syndrome score was conducted.Uterine volume was detected by color Doppler ultrasound on the day of enrollment and 6 months after treatment(avoiding the menstrual period).Levels of VEGF,CA199 and CA125 were detected by ELISA before treatment and 6 months after treatment.The adverse reactions and recurrence of patients were observed and recorded.Results The total effective rate was higher in the treatment group than that of the control group(P＜0.05).After treatment,the VAS score,TCM syndrome score,menstrual volume,uterine volume,serum levels of VEGF,CA199 and CA125 were lower in both groups than those before treatment,and the treatment group was even lower than those of the control group(P＜0.05).There was no statistically significant difference in the total incidence of adverse reactions between the two groups,and the recurrence rate of the treatment group was lower than that of the control group(P＜0.05).Conclusion The combination of Dangui Quyu pill and acupuncture has obvious effect and high safety in the treatment of ADS.

Objective:To investigate the effect and molecular mechanism of high concentration of IL-17A on osteoclast differentiation by inhibiting autophagy of osteoclast precursor cells through PI3K/Akt pathway.Methods:With RANKL (50 ng/ml) inducing osteoclast precursor cells (osteoclast we cells, OCPs), osteoclast differentiation model is set up. In osteoclast differentiation model of high levels of IL-17A (100 ng/ml), RAW264.7 cells were divided into negative control CTR-N group, CTR-R group with RANKL, IL-17A group, IL-17A+LY294002 group. BMMs were divided into negative control CTR-N group with M-CSF, CTR-R group, IL-17A group and IL-17A+LY294002 group with M-CSF and RANKL. IL-17A was applied to OCPs, and tartrate-resistant acid phosphatase (TRAP) staining was used to observe the number of osteoclast differentiation. The number of autolysosomes was observed under transmission electron microscope. RAW264.7 was treated with IL-17A. Western blot was used to detect the relative expression levels of p-Akt/Akt, p-mtor/mTOR, p-PI3K/PI3K, p-ULK1/ULK1, Cleaved-caspase3/caspase3, Beclin1/β-actin. The apoptosis rate of RAW264.7 cells treated with IL-17A was detected by flow cytometry. OCPs were treated with IL-17A and PI3K inhibitor LY294002, and TRAP staining was used to observe the number of osteoclast differentiation.Results:The TRAP staining showed that the positive ratio for RAW264.7 cells CTR-N group, CTR-R group, IL-17A group was 1.33%±0.58%, 100%±3.01%, 51.11%±4.02% with that of IL-17A significantly lower than CTR-R group ( t=16.970, P<0.05). The positive rates of BMMs in the CTR-N group, CTR-R group and IL-17A group were 1.67%±0.58%, 100%±1.01% and 50.33%±2.52%, respectively, with that of IL-17A group significantly lower than CTR-R group ( t=31.770, P<0.05). Transmission electron microscopy showed that the number of autophagosomes in RAW264.7 cells in CTR-R group and IL-17A group were 3.67±1.53 and 0.67±0.58, respectively, with significant difference between the groups ( t=3.182, P<0.05). While in BMMs cells CTR-R group and IL-17 the numbers of autophagosome were 3.00±1.00 and 0.33±0.58 with significant difference ( t=4.000, P<0.05); Western blot results showed 0.69±0.03、0.69±0.13、1.47±0.13、0.78±0.04、0.66±0.10、0.82±0.03 for RAW264.7 cells CTR-R group Akt/Akt, p-mTOR/mTOR, p-PI3K/PI3K, p-ULK1/ULK1, Cleaved caspase3/caspase3, Beclin1/β-Actin and 0.89±0.04、1.14±0.18、1.87±0.04、0.53±0.09、0.93±0.02、0.54±0.03 for RAW264.7 cells IL-17A group p-Akt/Akt, p-mTOR/mTOR, p-PI3K/PI3K, p-ULK1/ULK1, Cleaved caspase3/caspase3, Beclin1/β-Actin with significant difference ( t=6.708; t= 3.497; t=5.424; t=4.542; t=4.638; t=11.220, all P<0.05); Flow cytometry detection showed that in CTR-R group, IL-17A RAW264.7 cells apoptosis rates of group A were 6.92%±0.62%, 12.12%±0.69%, with significant difference between the two groups ( t=9.747, P<0.05); After using LY294002 TRAP staining, it showed a positive result of 9.00%±2.00%, 158.33%±3.51%, 100%±2.65% and 128.99%±4.01% for CTR-N, CTR-R, IL-17A and IL-17A+LY294002 in RAW264.7 cells respectively with significant difference between IL-17A+LY294002 group and the IL-17A in group A ( t=10.470, P<0.05). For BMMs cells CTR-N, CTR-R group, IL-17A in group, IL-17A+LY294002 group, the positive rate was 8.01%±0.99%, 151.67%±4.51%, 100%±3.61%, with significant difference between IL-17A+LY294002 group and IL-17A group ( t=6.535, P<0.05). Conclusion:High concentration of IL-17A inhibits osteoclast differentiation by inhibiting autophagy of osteoclast precursor cells through PI3K/Akt pathway.

Objective:To analyze the potential clinical biological factors influencing the major pathological response (MPR) to neoadjuvant immunochemotherapy in patients with resectable head and neck squamous cell carcinoma (HNSCC).Methods:This retrospective study enrolled patients with resectable HNSCC who underwent neoadjuvant immunochemotherapy at Sun Yat-sen University Cancer Center from June 1, 2019 to December 31, 2021. Binary logistic regression was used to analyze the correlation between clinical characteristics, inflammatory markers and MPR, and a nomogram model was constructed. The calibration curve and decision curve analysis were used to verify the predictive ability and accuracy of the nomogram model.Results:A total of 173 patients were included in the study, with 141 males and 32 females, aged from 22 to 83 years. After pathological assessment, the patients were divided into two groups: MPR group (108 cases) and non MPR group (65 cases). Logistics regression analysis indicated that the patients with HPV+oropharyngeal cancer, partial response or complete response by imaging assessment, low pre-treatment platelet/lymphocyte ratio, low pre-treatment C reactive protein/albumin ratio and lower pre-and post-treatment C reactive protein/albumin ratio difference were more likely to have MPR (all P<0.05). Nomogram model was constructed based on the above factors, with a C-index of 0.826 (95% CI: 0.760-0.892), and the calibration curve and decision curve analysis confirmed the prediction accuracy of the model. Conclusion:This study shows that many factors are related to MPR of patients with resectable HNSCC receiving neoadjuvant immunochemotherapy and the constructed nomogram model helps to develop personalized treatment strategies for the patients.

Objective:To construct the prediction model of knee degeneration in patients with knee osteoarthritis based on bioelectrical impedance index, and evaluate the prediction performance and application efficiency of the model.Methods:This was a cross-sectional study. From May to July 2021, 248 knee joints of 124 patients with knee osteoarthritis at home from Shijiazhuang Yuqiang Community Health Service Center who participated in physical examination were selected by convenience sampling to establish the model. According to Kellgren-Lawrence (K-L) grading system, the knee joints were divided into four groups, namely K-L1 ( n=19) , K-L2 ( n=103) , K-L3 ( n=96) , and K-L4 ( n=30) . The indicators included in the model were selected through analysis of variance or Kruskal-Wallis test, and a prediction model of knee degeneration was established using support vector machine, and the model was optimized using grid parameter optimization method. The prediction performance of the model was evaluated by the area under the receiver operating characteristic (ROC) curve, sensitivity, specificity, accuracy, positive predictive value and negative predictive value. Results:The indicators in the model included age, complications, lumbar/back/hip pain, high-risk occupation, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) -pain, WOMAC-function, capacitive reactance and phase angle. The area under the ROC curve of the training set model was 0.999, the prediction accuracy was 0.920, and the 95% confidence interval was 0.868 to 0.957. The area under the ROC curve of the test set model was 0.833, the prediction accuracy was 0.682, and the 95% confidence interval was 0.572 to 0.780.Conclusions:The prediction model of knee degeneration has good prediction performance and is easy to use, which can be used as a screening tool for knee degeneration in patients with knee osteoarthritis.

Objective:To evaluate the effects of different psychological interventions on depression and pain in patients with arthritis using the method of network Meta-analysis.Methods:Randomized controlled trials (RCTs) on psychological interventions for arthritis patients were systematically searched in PubMed, Embase, Cochrane Library, CINAHL, Web of Science, PsycINFO, WanFang and CNKI databases. The retrieval time limit was from the establishment of databases to September 30, 2021. The literature was screened according to inclusion and exclusion criteria, and quality was assessed using the Cochrane Manual recommended RCT bias risk assessment tool. STATA 15.0 software was used for network Meta-analysis according to frequency framework.Results:A total of 59 RCTS were included, involving 8 psychological interventions and 5 726 patients. For the primary outcome of depression, at post-intervention, the ranking results showed that relaxation was the most likely to be the best intervention. However, in the comparison of different intervention effects, only cognitive behavioral therapy showed statistically significant difference compared with the control group ( P<0.05) . At follow-up, the ranking results showed that cognitive behavioral therapy was the most likely to be the best, and the difference was statistically significant compared with the control group ( P < 0.05) . After intervention, for the secondary index pain, the ranking results showed that hypnosis intervention was the most likely to be the best, but the intervention effect of relaxation intervention and cognitive behavioral therapy was statistically significant compared with the control group ( P< 0.05) . At follow-up, the ranking results showed that the best possibility of receiving commitment therapy was the highest, and the difference was statistically significant compared with the control group ( P < 0.05) . Conclusions:Cognitive behavioral therapy has the best effect on depression indicators in arthritis patients, and relaxation intervention and acceptance commitment therapy have the best effects on pain indicators after intervention and at follow-up, respectively. The potential interventions include relaxation intervention and hypnosis intervention, which are worthy of further study.

Vitrectomy is the preferred surgical method for diabetic retinopathy, especially in the stage of proliferative diabetic retinopathy. Vitrectomy for diabetic retinopathy involves all aspects of vitrectomy, which is one of the signs of maturity of retinal surgeons. With the application of minimally invasive vitrectomy and perioperative anti-neovascularization drugs, indications and timing of surgery, perioperative medication, management of vitreous, and whether combined with cataract surgery have changed greatly, and new understanding is needed. Evidence-based clinical research on the timing of diabetic retinopathy surgery and perioperative drug use should be carried out to provide a new theoretical basis for the surgical treatment of diabetic retinopathy.

Objective:To observe the changes of macular morphology and blood flow after minimally invasive vitrectomy (PPV) in patients with severe non-proliferative diabetic retinopathy (sNPDR).Methods:A prospective clinical study. From January 2020 to April 2021, 17 consecutive sNPDR patients with 17 eyes who were diagnosed and received PPV treatment at the Zhongshan Ophthalmic Center of Sun Yat-sen University were included in the study. There were 12 males with 12 eyes and 5 females with 5 eyes; the average age was 55 years old; the average duration of diabetes was 11 years; the average glycosylated hemoglobin was 7.9%. Before the operation and 1, 3, and 6 months after the operation, all the affected eyes underwent best corrected visual acuity (BCVA), standard 7-field fundus color photography, and optical coherence tomography angiography (OCTA). An OCTA instrument was used to scan the macular area of the affected eye with in the range of 3 mm×3 mm to measure the central subfoveal thickness (CST), the thickness of the ganglion cell complex (GCC) in the macular area, the thickness of the retinal nerve fiber layer (RNFL), and the superficial capillary plexus (SCP) vessel density and perfusion density in the macular area, macular avascular zone (FAZ) area, a-circularity index (AI). Before the operation and 6 months after the operation, the least significant difference test was used for the pairwise comparison.Results:Before the operation, 1, 3, and 6 months after the operation, the FAZ area of the macular area were 0.34±0.14, 0.35±0.10, 0.37±0.10, 0.36±0.13 mm 2, respectively; AI were 0.52±0.13, 0.54±0.11, 0.57±0.10, 0.60±0.11; CST was 282.6±66.7, 290.4±70.9, 287.2±67.5, 273.2± 49.6 μm; GCC thickness were 77.1±15.5, 74.3±13.9, 72.6±16.2, 78.5±18.3 μm; the thickness of RNFL was 97.9±13.8, 101.3±14.6, 97.7±12.0, 96.1±11.4 μm, respectively. The overall blood flow density of SCP in the macula were (16.79±1.43)%, (16.71±1.82)%, (17.30±2.25)%, (17.35±1.22)%; the overall perfusion density were 0.32±0.02, 0.32±0.03, 0.33±0.03, 0.33±0.02, respectively. After the operation, the CST increased first and then decreased; the thickness of RNFL increased 1 month after the operation, and then gradually decreased. Comparison of the parameters before and 6 months after the operation showed that the AI improved, and the difference was statistically significant ( P=0.049); the difference in FAZ area and the thickness of CST, GCC, and RNFL was not statistically significant ( P=0.600, 0.694, 0.802, 0.712); There was no statistically significant difference in the retina SCP blood flow density and perfusion density in the macular area ( P=0.347, 0.361). Conclusion:Compared with before surgery, there is no significant change in macular structure and blood flow density in sNPDR patients within 6 months after minimally invasive PPV.

Objective:To evaluate the feasibility and clinical efficiency of robot-assisted laparoscopic radical prostatectomy (RARP) via extraperitoneal PORT-free single incision approach.Methods:The data of 33 patients with prostate cancer underwent the extraperitoneal PORT-free single incision RARP from November 2020 to January 2021 in Sichuan Provincial People's Hospital was retrospectively reviewed. The average age was 66.7 (58-78) years, the median PSA was 20.77 (2.89, 56.44) ng/m, and the mean Gleason score was 7.0 (6.0-9.0). The mean prostate volume was 48.4 (25.0-220.0) ml. Clinical stage: 32 cases was in cT 2a-2cN 0M 0, 1 case in cT 3aN 0M 0. 16 cases had a history of operation. All 33 operations were performed by the same operator. All operations were performed by extraperitoneal PORT-free single-incision approach. The surgical condition, postoperative complication, pathology, and follow-up results were observed. Results:In this study, 33 operations were successfully completed without conversion to open or additional single hole channel instruments. The average operation time was 61.3 (38.0-120.0) min, with the mean intraoperative bleeding volume of 72.2 (45.0-220.0) ml and the mean bladder neck urethral anastomosis time of 11.7 (8.5-15.7) min. The mean postoperative hospital stay was 7.9 (6.0-15.0) d, the mean postoperative indwelling time of urinary catheter was 6.8 (6.0-14.0) d, and the mean postoperative evacuation time was 1.0 (0.5-3.0) d. The average incision length was 5.2 (4.6-5.8) cm. There was no obvious complications. The postoperative pathological stage: 21 cases were in < pT 3a, 12 cases were in ≥ pT 3a, and 6 cases (18.8%) had positive resection margin. 29 cases (88.9%) acquired satisfactory urinary continence after operation, and the frequency of urinary pad use was ≤ 1 tablet/day. Conclusions:The extraperitoneal single-incision RARP surgical channel without PORT is safe and feasible with a satisfying cosmetic effect, which saves costs and requires less specific channel device. Simultaneously, the new approach has strong replicability, short-term tumor control and urinary control effect with rapid postoperative recovery. However, the sample size of this study is relatively small, which needs further research and demonstration