Objective To investigate the roles of RU486 inhibiting 3T3-L1 pre-adipocytes differentiation and regulating NF-κB activation. Methods Cells were treated with RU486 with concentrations of 0.1 ~ 10μmol/L for 48 h , then the relative contents of triglyceride were analyzed by Oil-Red O staining assay on 9 th day during adipogenesis. The mRNA expressions of PPARγ2,C/EBPa, LPL and FAS were further measured by Real-time PCR. IκBα protein level was detected by Western bolt and nuclear translocation of NF-κB was observed by immunofluorescence assay. Results The relative contents of triglyceride decreased with the increasing of RU486 concentration. Compared with the control, the relative contents of triglyceride in RU486-treatment groups from 0.5 μmol/L were significantly decreased (P < 0.05 or P < 0.01). Compared with the control, PPARγ2, C/EBPa, LPL and FAS mRNA expression and IκBα protein level were significantly decreased (P < 0.01) and NF-κB nuclear translocated from cytoplasm to nucleus in Group 5 mmol/L RU486. Conclusions RU486 could down regulate IκBα protein level , activate NF-κB nuclear translocation , then down regulate PPARγ2 , C/EBPa , LPL and FAS mRNA expression and inhibit adipocytes differentiation.

Objective To pan and characterize anti-HIV-1 Fab by the phage antibody library technology. Methods Total RNA were extracted from lymphocytes which were isolated from peripheral blood collected from asymptomatic HIV-1 infected donors with high titer antibody against HIV-1. The genes of heavy chains Fd fragment and light chains of antibody were amplified by RT-PCR. The phagmids pComb3X cloned Fd and light chain genes were transformed into E. coli XL1-Blue by electroporation to construct phage Fab library. By three runs of "absorption-elution-neutralization-enrichment", the clones were induced by IPTG and characterized by ELISA. The positive clones were sequenced and analyzed the sequences. Subsequently, Fab antibodies of these positive clones were induced to expressed and purified, then the recombinant virus neutralization assay was performed. Results A phage Fab library was constructed with 8×106 members, and 11 positive clones were obtained by detecting IPTG-induced-expressing Fabs with ELISA. By analysis of the sequences, 10 light chain genes and 8 Fd genes were ensured to be obtained. Compared with the genes of anti-HIV-1 antibodies in HIV sequence database, the gene sequences we obtained were highly homologous to some patent genes of anti-HIV-1 gp120 antibodies in HIV sequence database( light chains with 60%-90% identity, Fd with 71%-85% identity); The CDRs of these positive clones were determined by comparing the positive clone genes with antibodies' genes in V base database, furthermore, CDRH3 of these positive clones has the length of 12-22 aa. Strand shift had little effect to improving affinity of our Fab clones. Fab antibodies were induced to express at the concentration of ＞ 10 mg/L. Three Fab antibodies neutralize HIV-1 virus to some extent. Conclusion The studies will provide the basis on further study on the anti-HIV-1 Fabs obtained successfully.

Objective To investigate the feasibility of amplitude of intraoperative nerve action potentials (NAP) for early quantitative diagnosis of peripheral nerve injury. Methods The sciatic nerve injury model were established in 16 rabbits. Intraoperative NAP were recorded after 4 weeks. According to amplitude of NAP, the injuried nerve were divided into 3 groups: NAP ＜ 100 μV in A group, 100 μV ≤NAP ＜ 500 μV in B group, NAP ≥ 500 μV in C group. Nerve specimen 1cm distal to injuried point were resected that received glycine silver stain and image analysis including number, diameter and cross section area of regenerative axons. Footprint parameter and ulcer area were measured and contrasted between each two groups. Results The number, diameter and cross section area of A group regenerative axons have significant difference with B and C group, no significant difference between B and C group; Footprint parameter and ulcer area have significant difference in each two groups. Conclusion Amplitude of intraoperative NAP can be a quantitative criteria to diagnose the degree of peripheral nerve injury that provides experiment evidence for guide intraoperative decision-making in clinical practice.

Objective: To validation and optimization the indicator system of risk assessment for mechanical cuts. Methods: The risk assessment index system of mechanical cutting injury established earlier was used to assess the risk of mechanical cutting injury in 40 cases of mechanical cutting injury registered from January 2015 to December 2017 and 40 similar positions without accidents in the same period. The multiple stepwise regression analysis was used to screen the indicator system, and to adjust the weight coefficient of each index. The total coincidence rate and Kappa value were compared between before and after optimization respectively. Results: The new index system has 3 first-class indicators, 10 second-class indicators and 14 three-class indicators, fewer than the old index system which has 3 first-class indicators, 10 second-class indicators, 34 three-class indicators. There three indicators have revamped in the first-class. The total of coincidence rates of the new and old indicator systems were 67.50% and 90.00%, the difference was statistically significant (P<0.01). The Kappa value were 0.35 and 0.80, respectively. Conclusion The evaluation results with new indicator systems is more consistent with the actual hazard detection the the old indicator systems, and scientific, reasonable and practical, and the indicator system of risk assessment for mechanical cuts can be used for the risk assessment of mechanical cutting injuries.

Objective:To retrospectively analyze the characteristics and influencing factors of liver function changes in 111 elderly patients with COVID-19 pneumonia.Methods:111 elderly patients with COVID-19 admitted to the Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from February 5 to March 3, 2020 were enrolled. According to the severity of disease and liver function condition, they were divided into severe group ( n=40), normal group ( n=71), abnormal liver function group ( n=86) and normal liver function group ( n=25). The indexes related to liver function changes [total bilirubin (TBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and γ-glutamyl transferase (GGT)] and related influencing factors were analyzed. Results:Among 111 cases, 86 (77.5%) had abnormal liver function of varying degrees, and 28 (25.2%) had liver injury. The abnormal rates of TBil, AST, ALP and GGT were significantly higher in the severe group than normal group ( P<0.05). There were no significant differences in age, ribavirin, glucocorticoid and the application of lopinavir-ritonavir tablets between the abnormal liver function and the normal group ( P>0.05). The proportion of male was significantly higher in the abnormal liver function than normal liver function group ( P<0.05). Conclusion:Elderly COVID-19 patients have a higher proportion of abnormal liver function, and patients in the severe group are more likely to have higher level of TB, AST, ALP and GGT. The abnormal liver function may be related to the direct viral infection of the liver and the inflammatory immune response of the body after infection in elderly patients.

ObjectiveTo explore the clinical selection and application of cell suspension examination (CSE) or histopathological technique (HPT) in detecting sperm in the testis tissue obtained by testicular sperm aspiration (TESA) in patients with non-obstructive azoospermia (NOA).

METHODSTotally, 1 006 NOA patients underwent TESA and their testis tissues were subjected to CSE or HPT for sperm detection. Based on the results of CSE, the testicular tissue samples were divided into groups A (with sperm, n = 567) and B (without sperm, n = 439) and the results were compared with those of HPT.

RESULTSHPT showed 508 cases with but 59 without sperm in group A, and 403 with and 36 without sperm in group B. The consistency rate of CSE with that of HPT was 90.56% (Kappa =0.809), and CSE exhibited a significantly higher rate of sperm detection than HPT (56.36% vs 54.08%, P=0.023).

CONCLUSIONSCSE combined with HPT for detecting sperm in the testis tissue of NOA patients undergoing diagnostic TESA helps clinical diagnosis and treatment. The results of CSE have a decisive significance for assisted reproductive therapy, while those of HPT may provide some definite etiological evidence for drug therapy or surgery.

Since the concept of patient versions of guidelines （PVGs） was introduced into China， several PVGs have been published in China， but we found that there is a big difference between the concept of PVG at home and abroad， and the reason for this difference has not been reasonably explained， which has led to ambiguity and even misapplication of the PVG concept by guideline developers. By analyzing the background and purpose of PVGs， and the understanding of the PVG concept by domestic scholars， we proposed the term patient guidelines （PGs）. This refers to guidelines developed under the principles of evidence-based medicine， centered on health issues that concern patients， and based on the best available evidence， intended for patient use. Except for the general attribute of providing information or education， which is typical of common health education materials， PGs also provide recommendations and assist in decision-making， so PGs include both the patient versions of guidelines （PVG） as defined by the Guidelines International Network （GIN） and "patient-directed guidelines"， i.e. clinical practice guidelines resulting from the adaptation or reformulation of recommendations through clinical practice guidelines.

At present， the process and methodology of patient guidelines （PGs） development varies greatly and lacks systematic and standardised guidance. In addition to the interviews with PG developers， we have sorted out the relevant methodology for the adaptation and development of existing clinical practice guideline recommendations and facilitated expert deliberations to achieve a consensus， so as to finally put forward a proposal for guidance on the process and methodology for the development of PGs. The development of PGs can be divided into the preparation stage， the construction stage， and the completion stage in general， but the specific steps vary according to the different modes of development of PGs. The development process of Model 1 is basically the same as the patient version of the guideline development process provided by the International Guidelines Network， i.e.， team formation， screening of recommendations， guideline drafing， user testing and feedback， approval and dissemination. The developer should also first determine the need for and scope of translating the clinical practice guideline into a patient version during the preparation phase. Model 2 adds user experience and feedback to the conventional clinical practice guideline development process （forming a team， determining the scope of the PG， searching， evaluating and integrating evidence， forming recommendations， writing the guideline， and expert review）. Based on the different models， we sort out the process and methods of PG development and introduce the specific methods of PG development， including how to identify the clinical problem and how to form recommendations based on the existing clinical practice guidelines， with a view to providing reference for guideline developers and related researchers.

Objective To investigate the prevalence of Spirometra mansoni infections in hosts in Jiangsu Province, so as to provide the scientific basis for the management of sparganosis mansoni. Methods From 2018 to 2019, nine counties (cities, districts) were randomly selected from Jiangsu Province as the survey sites, and 100 healthy individuals were randomly selected to perform the serological test of S. mansoni infections and the detection of S. mansoni eggs. The procercoids were detected in the intermediate host Cyclops, and the S. mansoni eggs were identified in the stool samples of the definitive hosts cats and dogs. Results The prevalence of S. mansoni human infections was 0 (0/900) in the 9 survey sites of Jiangsu Province, and the sero-prevalence of the specific IgG antibody against S. mansoni was 1.22% (11/900). The positive rate of procercoids was 0.33% (3/900) in Cyclops. In addition, the S. mansoni egg-positive rate was 1.48% (2/135) in cats and dogs. Conclusions Sparganosis mansoni is prevalent in Jiangsu Province. Health education pertaining to the damages of sparganosis mansoni and the route of S. mansoni infections should be improved.