Objective To observe the changes in levels of plasma leukocyte cell-derived chemnotaxin 2 (LECT2) in patients with newly diagnosed type 2 diabetes (T2DM) complicated with non-alcoholic fatty liver disease (NAFLD),and to investigate the clinical significance.Methods A total of 137 subjects were enrolled in the study,including 50 patients with newly diagnosed T2DM complicated with NAFLD,47 patients with newly diagnosed T2DM with non-NAFLD and 40 healthy subjects.The level of plasma LECT2 was determined by ELISA.Anthropometric data and other biochemical indicators were measured in three groups.The body mass index (BMI) and waist-to-hip ratio (WHR) were calculated.Insulin resistance and pancreas β-cell function were determined by homeostasis model assessment (HOMA-IR,HOMA-％β).Results Plasma concentration of LECT2 in patients with newly diagnosed T2DM complicated with non-NAFLD was higher than that in type 2 diabetic patients with non-NAFLD [(32.95 ± 10.11 vs 29.08 ± 7.54) ng/mL,P ＜ 0.01].Plasma LECT2 levels in both groups were significantly higher than that in normal control group [(22.38 ± 4.40) ng/mL,P ＜ 0.01].Plasma LECT2 level was positively correlated with BMI,FPG,FINS,C peptide,HbA1c,GGT,TG and HOMA-IR,while negatively with HDL-C and HOMA-％ β (all P ＜ 0.01).Multivariate regression analysis showed that levels of BMI,FPG and HDL-C were important factors affecting plasma LECT2 level.Conclusions Plasma LECT2 concentration significantly elevates in patients with newly diagnosed T2DM complicated with non-NAFLD.Plasma level of LECT2 is closely correlated with insulin resistance and glycolipid metabolism.LECT2 may play an important role in the patho genesis of insulin resistance and T2DM.

Objective To quantitatively analyze the histological and MR images of repaired tissue (RT) following microfracture for knee joint osteochondritis dissecans (OCD) in rabbit models at different time points,make comparisons with the RT performances of joint debribement,explore the efficiency of the microfracture treatment for OCD.Methods Twenty-seven New Zealand rabbits were randomly assigned into 3 groups (sacrificed at the end of 3,5 and 7 weeks post-operation respectively),with 9 in each group.For each rabbit,one knee joint was made into an OCD model.In each group,6 were for microfracture treatment,and the other 3 were for joint debridement as control.MR scan,which mainly included sequences of 3D double echo steady state sequence (3D-DESS) and T2-mapping,was taken at 3,5 and 7 weeks postoperation.The thickness index and T2 value index of RT were calculated and T2-mapping of repaired region was drafted.Then the operation sites were removed to make histological sections of HE and Masson staining.The modified O' Driscoll score system was employed to make semi-quantitative evaluation for the histological performance of RT.Comparisons were made with respect to MR and histological findings between two treatments at each time point using unpaired Student t test.Effects of two treatments were evaluated longitudinally by comparing the results of three time points using one-way ANOVA.Results The post-operation thickness indexes of two groups increased gradually (F =33.940,28.841,P ＜ 0.05),T2 value indexes decreased (F =80.183,206.206,P ＜ 0.05),and O'driscoll scores increased gradually (F =29.867,17.167,P ＜ 0.05).At each time point,the thickness index of microfracture was higher than that of debridement group(3-week:0.743 ±0.048 vs 0.624 ±0.013,t =4.077 ;5-week:0.813 ±0.031 vs 0.734 ±0.015,t =4.107 ; 7-week: 0.972 ± 0.064 vs 0.777 ± 0.039,t =4.782 ; P ＜ 0.05),and the defects of microfracture in 7-week group were almost fully recovered while the defects of the debridement in 7-week group were not.The T2 value index of microfracture group was lower than that of debridement group at 3-week point(1.338 ±0.043 vs 1.510 ± 0.009,t =6.583,P ＜ 0.05),but it was higher than that of debridement group at 5-week and 7-week points (5-week: 1.284 ± 0.097 vs 1.116 ± 0.068,t =2.663 ; 7-week: 0.916 ± 0.036 vs 0.843 ± 0.016,t =3.283 ; P ＜ 0.05).The O'Driscoll score of microfracture group was higher than that of joint debridement at every time point(3-week:7.167 ±0.753 vs 4.667 ±0.577,t =5.000;5-week: 9.833 ± 1.169 vs 7.667 ± 0.577,t =2.960 ; 7-week: 11.167 ± 0.753 vs 8.333 ± 1.155,t =4.520 ; P ＜ 0.05).For microfracture group,the RT was mainly repaired by fibrocartilage and got matured gradually with more production of well-distributed collagen fibrils ; while for joint debridement group,the RT was mainly repaired by fibrous and scar tissue.Conclusions The post-operation repairing thickness and tissue composition of microfracture for OCD are superior to that of joint debridement.MR 3D-DESS and T2-mapping can show the thickness and tissue composition of the RT after OCD treatments,provide effective evaluation of repairing conditions,and they are of great importance on the OCD post-op follow-up.

Objective Through the analysis of the pathogens distribution of out-patient department and emergency department patients with diarrhea from Shenzhen Hospital of Peking University in 2013~2014,so as to provide reference for the clinical diagnosis and treatment of diarrhea disease.While understanding the improved Salmonella detection results.Methods Var-ied pathogenic bacteria were isolated and identified from 1 719 diarhea stool samples of native Shenzhen Hospital of Peking University from 2013 to 2014 through enrich culture,separate culture,biochemistry,serology etc.Pathogenic virus were test-ed for 451 watery stool specimens by fluorescence PCR.Analyzed statistical differences between the direct inoculation and selenite cystine broth enrichment for Salmonella.Results Picked out 143 disease germs from 1 719 examples diarrhea pa-tient’s stool samples,among which there were 25 strains of ETEC,12 strains of EPEC,8 strains of EAEC,1 strains of EIEC,19 strains ofVibrio parahemolyticus,76 strains Salmonella,2 strains Shigella and 0 strain ofVibrio cholera.There were 10 samples with two disease germs timely.Picked out 189 disease viruses from 451 examples diarrhea patient’s stool without disease germs,among which there were 79 Rotaviruspositive,91 Norwalkvirus positive,9 Adenoviruspositive,10 Astrovirus positive.There were 4 samples with Rotavirus and Norwalkvirus timely.After Salmonella ways to improve the positive rate of 0.6% (17/2 627)increased to 4.4% (76/1 719),χ2=67.2,P<0.01,the difference was statistically signifi-cant.Conclusion The detectable rate of Salmonella and Norwalk virus was the majority,and the clinic enhance the test of the diarrhea pathogenic microorganism,including the improvement of detection method,to reduce the missing rate of them,to provide the scientific basis for the diagnostical therapectic measures.

Objective To investigate the effects of AGEs-RAGE on the apoptosis of GLUTag cells and explore the possiblc mechanism.Methods GLUTag cells treated with 0、100、200、300μg/ml of AGEs for 24h were examined for gene and protein expression of RAGE using RT-PCR and western blotting,respectively.GLUTag cells were randomly divided into four groups:control,200μg/ml AGEs,AGEs+siRNA-RAGE and AGEs+apocynin.The protein expression of p22phox、p47phox 、Bcl-2、Bax in the cells were detected with western blotting.The reactive oxygen species (ROS) levels were examined using 2'7'-dichlorodihydroflur-rescein diacetate (DCFH-DA) and the apoptosis of L cells were tested by AnnexinV-FITC/PI.Results AGEs increased thc cxpression of RAGE in a dose dependent manner.Treatment with AGEs induced a significant increase in the expression of p22phox,p47phoxand the activity of ROS,caused up-regulation of Bax and down-regulation of Bcl-2,which enhanced the apoptosis of GLUTag cells.Apocynin,the inhibitor of NADPH oxidase,prevented those responses and the effects caused by AGEs were abolished by inhibition of RAGE activity with siRNA.Conclusion AGEs positively regulate the exprcssion of NADPH oxidase-derived ROS and its down-steam signaling pathway p53/Bax by targeting RAGE,leading to the apoptosis of GLUTag cells.

The incidence rate of primary liver cancer continues to increase around the world, with a younger age of onset and poorer prognosis. As the most classic regulator of cell polarity and density, mechanical signal transduction, cell proliferation, and organ development, the Hippo pathway can promote the development and progression of various cancers including primary liver cancer. YAP, a classic nuclear effector of the Hippo pathway, is significantly upregulated in primary liver cancer and promotes the development of drug resistance. This article aims to investigate the association of the dysregulation of the Hippo signaling pathway with the development and progression of primary liver cancer and analyzes the mechanism of action of the Hippo signaling pathway in the drug resistance of primary liver cancer as an early event of the development of primary liver cancer, which is of great significance for exploring new treatment strategies for primary liver cancer.

OBJECTIVETo observe the effects of Tripterygium polyglycosides (TWP) on mucous immune function in rat models of arthritis induced respectively with collagen-II induced arthritis (CIA) & adjuvant arthritis (AA).

METHODSCIA and AA model rats were induced by immunization with collagen II emulsified with complete Freund's adjuvant and complete Freund's adjuvant respectively and treated with TWP. Rats' mucus, systemic immunological indexes (peripheral subsets of T cells), local inflammatory factors (IL-6, TNF-alpha, COX-2,and NF-kappaB, etc. ) were observed.

RESULTSIn CIA model group, CD4+ in Peyer's Patch (PP), peripheral CD4+ and CD8+ positive T cells all raised, while in the AA model group, CD4+ lowered and CD8+ raised on PP, with both subsets increased. Effects of TWP on T lymphocyte subsets in PP and blood of the two models were different. High leveled IL-6, TNF-alpha, COX-2 and NF-kappaB expression could be seen in both model groups, and these inflammatory media could be inhibited by TWP.

CONCLUSIONThere exist similarities and differences between the two models in aspects of mucus immune response and effect of TWP on them.

Animals ; Arthritis, Experimental ; chemically induced ; drug therapy ; immunology ; Collagen Type II ; Freund's Adjuvant ; Glycosides ; pharmacology ; Male ; Mucous Membrane ; immunology ; Rats ; Rats, Sprague-Dawley ; Tripterygium ; chemistry

ObjectiveTo investigate the effect of the Yap1 gene and tanshinone ⅡA on the proliferation, migration, and invasion abilities of Huh-7 hepatoma cells. MethodsA total of 10 pairs of human hepatocellular carcinoma (HCC) samples and adjacent tissue samples were collected in Zhongnan Hospital of Wuhan University from June 1 to December 1, 2019. Quantitative real-time PCR and Western blotting were used to measure the expression of the Yap1 gene and phenotype-related molecules. MTT cell proliferation detection reagent was used to measure the inhibition rate of cell proliferation after the treatment with different concentrations of tanshinone ⅡA. Western blotting was used to measure the changes in the expression of apoptosis-and migration-related markers after different interventions. Flow cytometry and Transwell assay were used to measure apoptosis and cell migration and invasion abilities. The data of 375 cases of liver cancer and 50 cases of relatively normal liver tissue samples were downloaded from The Cancer Genome Atlas, including clinicopathological information. The t-test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. ResultsIn 8 of the 10 pairs of HCC samples and adjacent tissue samples, HCC samples had significantly higher expression of Yap1 than the adjacent tissue samples. Compared with the normal human liver epithelial cells L02, the Huh-7 and HCCL-M3 hepatoma cells had a significant increase in the expression of Yap1. The silencing efficiency of si-Yap1-3 transfection reached 87.004% at the protein level. MTT results showed that tanshinone ⅡA effectively inhibited the proliferation of Huh-7 cells, with a half inhibitory concentration of 8.683 μmol/L. After the cells were treated with si-Yap1-3 and tanshinone ⅡA, there was an increase in the expression of the downstream marker for proliferation and migration E-cadherin and a reduction in the expression of vimentin, and the results of Transwell assay showed that compared with the si-NC group, the tanshinone ⅡA+si-Yap1-3 group had significant reductions in the migration and invasion abilities of Huh-7 cells (migration: 43.19±2.88 vs 132.20±10.03, t=8.527, P=0.001; invasion: 53.95±4.20 vs 179.10±11.11, t=4.484, P=0.011). The group treated with si-Yap1-3 and tanshinone ⅡA had an increase in the expression of the apoptosis-related marker Bax and a reduction in the expression of Bcl-2, as well as a significantly higher early apoptosis rate than the si-NC group (2598% vs 9.21%, χ2=4.078, P＜0.05). ConclusionOncogene Yap1 silencing combined with tanshinone ⅡA can promote the apoptosis of Huh-7 hepatoma cells and inhibit their migration and invasion, which can provide certain guiding significance for clinical medication.

Objective:To compare the efficacy of arthroscopic all-inside ligament repair with suture augmentation and ligament reconstruction with tendons in the treatment of chronic ankle instability with poor remnant quality of the anterior talofibular ligament (ATFL) tissue.Methods:A retrospective cohort study was conducted to analyze the clinical data of 37 patients with chronic ankle instability treated at Huashan Hospital Affiliated to Fudan University from January 2018 to August 2020, including 34 males (34 ankles) and 3 females (3 ankles); aged 18-57 years [(32.2±7.2)years]. The time from injury to operation ranged from 3-360 months [48(12, 120)months]. All patients underwent arthroscopic all-inside ankle stabilization surgery, of which 19 underwent ligament repair with suture augmentation (augmented repair group) and 18 underwent traditional ligament reconstruction with allograft/autograft tendons (tendon reconstruction group). The degree of ATFL injury and intra-articular lesions (osteophytes, loose bodies and cartilage damage) were recorded during the operation. The Karlsson scale and Tegner scale were evaluated before operation and at the last follow-up. The number of patients who were able to complete partial weight-bearing/return to normal walking/return to sports postoperatively and the time required were compared between the two groups. Postoperative complications were observed.Results:All patients were followed up for 12-32 months [21(16, 28)months]. There were no significant differences in the degree of ATFL injury and intra-articular lesions (osteophytes, loose bodies and cartilage damage) seen during the operation between the two groups (all P>0.05). At the last follow-up, the Karlsson score in augmented repair group and tendon reconstruction group [95.0(90.0, 98.5)points and 95.0(87.8, 99.3)points] was significantly higher than the preoperative level [65.0(51.0, 75.0)points and 65.0(53.3, 78.0)points] (all P<0.01). At the last follow-up, the Tegner score in augmented repair group and tendon reconstruction group [5.0(3.5, 6.0)points and 5.0(3.3, 6.0)points] were significantly higher than the preoperative level [3.0(2.0, 4.0)points and 2.5(1.3, 4.0)points] (all P<0.01). There were no significant differences in Karlsson score and Tegner score between the two groups (all P>0.05). All patients completed partial weight-bearing after 3.0(2.0, 4.0)weeks in augmented repair group and 4.0(3.5, 6.0)weeks in tendon reconstruction group. All patients returned to normal walking after 8.0(6.0, 9.0)weeks in augmented repair group and 8.0(5.5, 12.0)weeks in tendon reconstruction group. A total of 13 patients (63%) in augmented repair group and 13 patients (72%) in tendon reconstruction group successfully returned to sports postoperatively and the time required was 6.0(3.5, 8.0) months and 6.0(4.5, 12.0)months, respectively. There were no significant differences in the above indicators between the two groups (all P>0.05), but augmented repair group had a trend of faster completion of partial weight-bearing than tendon reconstruction group. There was 1 patient [5%(1/19)] in augmented repair group and 1 patient [6%(1/18)] in tendon reconstruction group who reported feelings of instability during exercise postoperatively ( P>0.05). None of the patients in augmented repair group had limited ankle range-of-motion, not different from 1 patient [6%(1/18)] in tendon reconstruction group ( P>0.05). Conclusion:In the treatment of chronic ankle instability with poor remnant quality of the anterior talofibular ligament (ATFL) tissue, both arthroscopic all-inside ligament repair with suture augmentation and ligament reconstruction with tendons can improve the short-term postoperative ankle function and activity level of the patients, and the former one has advantages such as simple operative procedures and none use of grafts.

Objective To construct an intervention program for diabetic retinopathy(DR)patients undergoing surgery based on the theory of'Timing It Right'.Methods Guided by the theory of'Timing It Right',based on literature review and semi-structured interview,the first draft of the intervention program was formulated.From December 2022 to February 2023,15 medical and nursing experts in ophthalmology and endocrinology from 5 provinces(cities)in Zhejiang,Sichuan,Shanxi,Chongqing and Shanghai were interviewed by Delphi expert for 2 rounds to seek their advice and revision,and the final draft was formed.Results The effective recovery rate of 2 rounds of expert consultations was 93.75％and 100％.The authority coefficients were 0.87.The importance and feasibility of Kendall'W coordination coefficients were 0.325,0.138 and 0.193,0.141 in 2 rounds,respectively(P＜0.001).The finalized intervention program consisted of 6 months,which included 5 first-level items,18 second-level items and 43 third-level items.Conclusion The intervention program for DR patients undergoing surgery based on the theory of'Timing It Right'is reliable,scientific,feasible and practical,and it can meet the needs of such patients,providing a reference for improving their self-management ability,improving their negative emotions and delaying disease progression.

Objective:To provide consistent data basis for the application of reference intervals for children blood cell analysis in different testing systems.Methods:According to the requirements of American Institute for Clinical and Laboratory Standardization (CLSI) EP9-A3 document, 45 samples were collected and Sysmex XN20-A1 were used as reference system. Beckman DxH800, Siemens ADVIA 2120i, and Mindray BC5310 were comparison systems. Complete blood count and leukocyte classification were performed by four systems. The outliers of the detection results were tested by the generalized extreme student deviate (ESD) method. An optimal regression model was selected by scatter diagram, deviation diagram and frequency distribution diagram, which was used to fit the regression equation and calculate the deviation at the medical decision level and reference interval. The acceptable range for blood count deviation was cited from the Analytical Quality Specifications for Routine Tests in Clinical Hematology. The acceptable range for leukocyte classification was based on the EQA program of Royal College of Pathologists of Australasia (RCPA).Results:After the outliers were deleted, the scatter plot showed a linear relationship between the reference system and the three comparison systems. The deviation plot showed that the differences were variable. Deming regression or Passing-Bablok regression was selected according to the data distribution. The determination coefficient R2 of reference system and three comparison systems ranged from 0.95 to 0.99 in blood count and leukocyte classification. At the upper and lower limits of the reference interval, the deviations between XN-20A1 and ADVIA 2120 system were all acceptable, except for MONO# at 0.12×10 9/L. The deviations of all parameters at medical decision level were within acceptable ranges. The lower limit of PLT is partially unacceptable at the level of medical decision related to treatment and prognosis. Conclusions:The results of complete blood count and leukocyte classification in reference system and the comparison system had good consistency within the children′s reference interval. Our study provided a scientific basis for the feasibility of adopting a unified reference interval for different detection systems.