Objective:To prepare rabbit polyclonal antibodies against respiratory syncytial virus (RSV) N protein and use them as the detection antibodies to establish a N-ELISA-based method for rapid detection of neutralizing antibodies.Methods:A plasmid of pET30a-N for the expression of RSV N protein was constructed. After purification, the protein was immunized into New Zealand rabbits to prepare polyclonal antibodies, which were used as the detection antibodies. Positive serum samples were diluted and used to neutralize RSV (100 TCID 50/well). Hep-2 cells were inoculated and cultured, and then the cells were fixed with 80% acetone. ELISA was performed to detect RSV N protein in infected cells. When the absorbance value of a well was below the cut-off value, it was regarded as the positive well in the neutralization test. The highest dilution of a positive well serum was the neutralizing antibody titer. After optimizting the antibody dilution, detection time, cell density and the duration of neutralization, the method for neutralizing antibody detection was established based on N-ELISA. The established method was verified by analyzing the influences of different cell generations and edge effects, and calculating the accuracy, repeatability and precision. The correlation between the established method and microneutralization method was analyzed by detecting human RSV IgG-positive serum. Results:The plasmid pET30a-N was successfully constructed, and the expressed N protein showed high purity and good specificity. After the third immunization, the antibody titer in rabbit serum was 1∶51 200, and the antibodies could specifically bind to RSV. The prepared rabbit anti-RSV N polyclonal antibodies had a titer of 1∶51 200, and showed good specificity. The neutralizing antibodies could be detected on day 4 with the established method, and the duration of neutralization was shortened to 30 min. Cell generations and the position of wells in the 96-well plate (edge well and non-edge well) had no significant effect on the method, and the repeatability, precision and accuracy of the method were good. In the detection of 64 RSV IgG-positive human serum samples by the established method and microneutralization method, the correlation coefficient was 0.929 6, indicating a good positive correlation between the two methods.Conclusions:A N-ELISA-based method for rapid neutralizing antibody detection is successfully established, which can be used to evaluate the serum antibody level after RSV vaccination.

The article summarizes the clinical experience of Professor JIA Chunsheng in treating cubital tunnel syndrome with various traditional Chinese medicine therapies,including superficial point-toward-point auricular acupuncture,ordinary acupuncture,fire-needle therapy,and oral Chinese medication,to inherit his academic characteristics,such as meridian-identified and stage-identified treatments,stressing the patient's body constitution and state,and emphasizing the holistic treatment,and to provide references for the popular science education and clinical treatment of cubital tunnel syndrome.

Objective To detect 17β-hydroxysteroid dehydrogenase type 1(17β-HSD1)expression in malignant epithelial ovarian tumors and analyze its relationship with pathological features and prognosis.Methods The clinical data of 70 patients with malignant epithelial ovarian tumors,70 with borderline epithelial ovarian tumors,and 70 with benign epithelial ovarian tumors were retrospectively analyzed.Immunohistochemistry was used to detect 17β-HSD1 expression.The difference in 17β-HSD1 expression levels among three groups were compared,and the relationship between 17β-HSD1 expression and various clinicopathological features and prognosis was ana-lyzed.Results Malignant epithelial ovarian tumor tissues had high 17β-HSD1 expression that was closely related to FIGO stage,lymph node metastasis,and carbohydrate antigen 125(CA125)and human epididymis protein 4(HE4)levels(P＜0.05),with no significant cor-relation with menopausal status,tumor differentiation degree,pathological type,and ascites(P＞0.05).The 17β-HSD1 expression levels,preoperative CA125 and HE4 levels,tumor differentiation degree,and FIGO stage affected the prognosis of malignant epithelial ovarian tumors.Multivariate Cox regression analysis suggested that the 17β-HSD1 expression levels and FIGO stage were independent risk factors for malignant epithelial ovarian tumor.Conclusion Malignant epithelial ovarian tumors had high 17β-HSD1 expression,which is closely related to the occurrence and development of tumors and among the independent risk factors affecting the overall survival.

Objective:To compare the clinical efficacy and safety of intra-articular platelet rich plasma combined with hyaluronic acid and platelet rich plasma alone in the treatment of knee osteoarthritis.Methods:The relevant literatures published between January 2010 and December 2021 on the randomized control study of intra-articular injection of platelet rich plasma and hyaluronic acid and injection of platelet rich plasma only in the treatment of knee osteoarthritis were searched. The bias risk of all the literatures included in the study was evaluated by Revman 5.3 software, and the data were processed and analyzed by Stata 16.0 software. The standardized mean difference ( SMD) was calculated for the difference of efficacy indexes, and the difference was compared by t- test. The odds ratio ( OR) was calculated for the difference of safety in-dex, and the difference was compared by t- test. Results:①A total of 7 relevant research literatures were in-cluded, all of which were in English. ②A total of 675 patients were included in the study, of which 330 patients were treated with intra-articular injection of platelet rich plasma and hyaluronic acid, and 345 patients were treated with intra-articular injection of platelet rich plasma only. ③The VAS scores of patients with platelet rich plasma and hyaluronic acid injection combined with injection and platelet rich plasma injection alone were compared. After 1 and 3 months of injection, the VAS scores of patients with platelet rich plasma combined with hyaluronic acid injection were compared with those of patients with platelet rich plasma injection only. There was no significant difference[ SMD(95% CI)=-0.02(-0.30, 0.27), t=-0.12, P=0.902; SMD(95% CI)=-0.40(-0.80, 0.00), t=-1.97, P=0.051]. After 6 months of injection, the VAS scores of patients with combined injection of platelet rich plasma and hyaluronic acid were significantly lower than those of patients with injection of platelet rich plasma only, and the difference was statistically significant [ SMD(95% CI)=-0.33 (-0.55, -0.11), t=-2.98, P=0.012]. ④The WOMAC scores of patients with combined injection of platelet rich plasma, hyaluronic acid and platelet rich plasma only were compared. After 12 months of treatment, the WOMAC scores of patients with combined injection of platelet rich plasma and hyaluronic acid were significantly higher than those of patients with PRP only, and the difference was statistically significant [ SMD(95% CI)=-0.30(-0.58, -0.03), t=-2.14, P=0.033]. ⑤The incidence of adverse events of platelet rich plasma combined with hyaluronic acid was lower than that of platelet rich plasma only [ OR(95% CI)=0.55(0.33, 0.93), P=0.021]. Conclusion:Compared with intra-articular injection of platelet rich plasma only, the short-term clinical efficacy of combined injection of platelet rich plasma and hyaluronic acid is equivalent to that of injection of platelet rich plasma only, but the long-term clinical efficacy is significantly better, and the safety of combined injection of platelet rich plasma and hyaluronic acid is significantly better.

Objective:To observe the long-term efficacy and late adverse reactions of simultaneous integrated boost intensity-modulated radiotherapy (SIB-IMRT) for mediastinal lymph node recurrence after radical surgery for esophageal squamous cell carcinoma (ESCC).Methods:A total of 20 ESCC patients with mediastinal lymph node recurrence (≤5) after radical surgery admitted to Department of Radiotherapy, Shanghai Ruijin Hospital between June 2019 and December 2021 were enrolled in this prospective study. Among them, 10 patients were enrolled in phase I study and 10 patients in phase II study. Four, 3 and 13 patients received three different doses of SIB-IMRT at 58.8 Gy/28 fractions, 64.4 Gy/28 fractions and 70.0 Gy/28 fractions for recurrent lesions, respectively. The overall survival (OS) rate, local control rate (LCR) and progression-free survival (PFS) were calculated by Kaplan-Meier analysis. Adverse reactions were also analyzed.Results:The most common sites of recurrence were 2R and 4 L, accounting for 35% and 25%, respectively. The median follow-up time was 32 months. For patients who received salvage chemoradiation after relapse, the 1-, 2- and 3-year OS rates were 100%, 88% and 78%, the 1-, 2- and 3-year PFS rates were 85%, 78% and 78%, respectively. The most common hematological toxicities were leukocytopenia and anemia. The most common nonhematological toxicity was esophagitis. However, no grade 3 or above esophagitis, pneumonia and cardiotoxicity were found. Three patients who received SIB-IMRT at 58.8 Gy/28 fractions died of distant metastases at 2 years after treatment, and 1 patient who received SIB-IMRT at 70.0 Gy/28 fractions died of distant metastases at 16 months after treatment.Conclusion:Salvage chemoradiotherapy using SIB-IMRT is efficacious and safe for mediastinal lymph node recurrence in ESCC patients after radical resection.

Objective:To explore the relationship between stigma, psychological resilience and quality of life in patients with stable schizophrenia.Methods:This study is a cross-sectional study. From January to December 2021, 305 patients with stable schizophrenia who were treated in the Outpatient Department of the Affiliated Brain Hospital of Nanjing Medical University were selected by convenient sampling. The survey was carried out by using the General Information Questionnaire, the World Health Organization Quality of Life-Brief Form (WHOQOL-BREF) , the Connor-Davison Resilience Scale (CD-RISC) , and the Perceived Devaluation Discrimination (PDD) . Pearson correlation analysis was used to analyze the relationship between quality of life, stigma and psychological resilience of schizophrenia patients. The structural equation model was established using AMOS 25.0 and the intermediary effect was verified. A total of 305 questionnaires were distributed and 286 valid questionnaires were recovered, with an effective recovery rate of 93.8% (286/305) .Results:Among 286 patients with stable schizophrenia, the scores of psychological resilience, stigma and quality of life were (56.49±14.28) , (29.62±5.70) and (63.15±11.67) respectively. There was a positive correlation between the psychological resilience and the quality of life ( r=0.461, P<0.01) , a negative correlation between the stigma and the quality of life ( r=-0.328, P<0.01) , and a negative correlation between the stigma and the psychological resilience ( r=-0.309, P<0.01) . The results of intermediary effect analysis showed that psychological resilience played a part of intermediary role between the stigma and the quality of life, and the indirect effect was statistically significant (β=- 0.167, P<0.001) , the intermediary effect accounted for 32.2% of the total effect. Conclusions:Psychological resilience is an intermediary variable between the stigma and the quality of life in patients with stable schizophrenia. Nurses should carry out intervention strategies based on the impact of psychological resilience path to reduce the patients' stigma and improve their quality of life.

Due to the virtues of no stutter peaks, low rates of mutation, and short amplicon sizes, insertion/deletion (InDel) polymorphism is an indispensable tool for analyzing degraded DNA samples from crime scenes for human identifications (Wang et al., 2021). Herein, a self-developed panel of 43 InDel loci constructed previously by our group was utilized to evaluate the genetic diversities and explore the genetic background of the Han Chinese from Beijing (HCB) including 301 random healthy individuals. The lengths of amplicons at 43 InDel loci in this panel ranged from 87 to 199 bp, which indicated that the panel could be used as an effective tool to utilize highly degraded DNA samples for human identity testing. The loci in this panel were validated and performed well for forensic degraded DNA samples (Jin et al., 2021). The combined discrimination power (PD) and combined probability of exclusion (PE) values in this panel indicated that the 43 InDel loci could be used as the candidate markers in personal identification and parentage testing of HCB. In addition, population genetic relationships between the HCB and 26 reference populations from five continents based on 19 overlapped InDel loci were displayed by constructing a phylogenetic tree, principal component analysis (PCA), and population genetic structure analysis. The results illustrated that the HCB had closer genetic relationships with the Han populations from Chinese different regions.
Beijing ; China ; Forensic Genetics/methods* ; Gene Frequency ; Genetics, Population ; Humans ; INDEL Mutation ; Phylogeny

Objective:Our study aims to determine histological regression and clinical improvement after long-term antiviral therapy in hepatitis B virus-related cirrhosis patients.Methods:Treatment-na?ve chronic hepatitis B patients with histologically or clinically diagnosed liver cirrhosis were enrolled. Liver biopsies were performed after 5 years entecavir-based antiviral treatment. Patients were followed up every 6 months. Cirrhosis regression was evaluated based on Metavir system and P-I-R score. Clinical improvement was evaluated before and after the long-term treatment. Kruskal Wallis test and Wilcoxon signed-rank test were used for continuous variables, Fisher's exact test was used for categorical variables and multivariate analysis was performed using logistic regression analysis.Results:Totals of 73 patients with HBV-related liver cirrhosis were enrolled. Among them, 30 (41.1%) patients were biopsy proved liver cirrhosis and the remaining 43 (58.9%) cirrhotic patients were diagnosed by clinical features. Based on Metavir system and P-I-R score, 72.6% (53/73) patients attained histological regression. Furthermore, 30.1% (22/73) were defined as significant regression (Metavir decrease ≥2 stage), 42.5% (31/73) were mild regression (Metavir decrease 1 stage or predominantly regressive by P-I-R system if still cirrhosis after treatment) and 27.4% (20/73) were the non-regression. Compared to levels of clinical characteristics at baseline, HBV DNA, ALT, AST, liver stiffness(decreased from 12.7 to 6.4 kPa in significant regression, from 18.1 to 7.3 kPa in mild regression and from 21.4 to 11.2 kPa in non-regression)and Ishak-HAI score significantly decreased after 5 years of anti-HBV treatment, while serum levels of platelets and albumin improved remarkably ( P<0.05). In multivariate analysis, only the pre-treatment liver stiffness level was associated with significant regression ( OR=0.887, 95% CI: 0.802-0.981, P=0.020). Conclusions:After long-term antiviral therapy, patients with HBV-related cirrhosis are easily to attain improvements in clinical parameters, while a certain percentage of these patients still cannot achieve histological reversal.

This paper reviews the concept, theoretical background, basic concepts, core technologies, specific procedures and application effects of solution-focused brief therapy in patients with depression, in order to provide a basis for medical staff to carry out research on solution-focused patients with depression.

[Abstract] Objective: To investigate the effect of HMGB1 gene on the growth of human epithelial ovarian cancer xenografts in nude mice, and to lay a foundation for finding new targets for the treatment of ovarian cancer. Methods: Human epithelial ovarian cancer SKOV3 cells in logarithmic growth phase were selected to establish a human epithelial ovarian cancer xenograft model in nude mice. Nude mice with successful model establishment were randomly divided into control group and HMGB1-siRNA group. On the 7th, 9th, 11th, 14th, and 16th days after cell inoculation, the same amount of saline and HMGB1-siRNA were respectively injected into two groups of mice under the armpit.After 3 weeks, the nude mice were sacrificed by cervical dislocation, the tumor tissues were separated, and the volume of the tumor was measured. The apoptosis of transplanted tumor cells was detected by Tunnel staining. The expressions of HMGB1, STAT3 and p-STAT3 were detected by Western blotting. The expression of vascular endothelial growth factorA(VEGF-A) and microvascularization were detected by immunohistochemistry. Results: Compared with the control group, the growth of tumor volume slowed down in HMGB1 siRNA group, and on the 21st day, the tumor volume of HMGB1-siRNA group was significantly smaller than that of the control group (P<0.05). HMGB1-siRNA successfully knocked down the expression of HMGB1 mRNA in transplanted tumor tissue. The apoptosis rate of tissue cells in HMGB1-siRNA group was significantly increased ([34±8]% vs [6±2]%, P=0.04), and the expressions of HMGB1 and p-STAT3 were significantly reduced (P<0.05). The expression of VEGF-Aand the number of microvessels were significantly lower than those of the control group (both P<0.05). Conclusion: Knockdown of HMGB1 gene reduces the expression of VEGF-A and microvessel formation possibly by inhibiting the HMGB1/STAT3 signaling pathway, thereby promoting the apoptosis of tumor tissues and slowing the growth of xenografts.