Objective To explore the feasibility and rationality of advanced integrated two-stage laparoscopic simulation training course in standardized training of surgical residents.Methods From December 2019 to December 2021,the advanced integrated two-stage laparoscopic simulation training course was carried out among 2019-2020 surgical residents who received standardized training in our hospital.The course was divided into two stages.In the first stage,BEST(best essential surgical technology training)course,adopted Darwin? endoscopic training system,Tianyan? endoscopic training system,Microport? 3D laparoscopic training system and simple simulative models were used.The second stage,BEST PLUS course,same platform as that in BEST course and in vitro animal models were used.The questionnaire survey method(before and after class questionnaire)was adopted to evaluate the curriculum setting,such as curriculum form,simulators,teaching method,time arrangement,curriculum difficulty,training effect,curriculum satisfaction and so on.Results A total of 37 surgical residents completed the two-stage course training and the questionnaire survey.The overall satisfaction rate with the curriculum setting was 100%.There were 32 residents(86.5%)thought that first stage training course could significantly improve their clinical skills,35 residents(94.6%)thought that second stage training course could significantly improve their clinical skills,and 36 resident(97.3%)thought that the first stage curriculum could significantly help them improve performance in the second stage curriculum.Conclusions The trainees had a high degree of recognition and satisfaction for the advanced integrated two-stage laparoscopic simulation training course.The overall design of course was reasonable and feasible,and was attractive to trainees.

Objective:To preliminarily explore the zoning of inflammatory lesions in patients with acute necrotizing pancreatitis (ANP) based on the peripancreatic membrane anatomy, and its impact on treatment outcome of ANP.Methods:Clinical data of 197 patients with ANP treated at Hunan Provincial People's Hospital from January 2021 to June 2022 were retrospectively analyzed, including 133 males and 64 females, aged (47.2±13.3) years old. Basic information, characteristics of pancreatitis, and imaging data were collected. The inflammatory lesions were partitioned based on the peripancreatic membrane anatomy. Patients were followed-up via outpatient visits or telephone reviews. According to the prognosis, patients were divided into the poor-prognosis group ( n=93), including patients with postoperative multi-organ failure, severe local complications, and death; and the non-poor-prognosis group ( n=104), including patients without these adverse outcomes. Multivariate logistic regression analysis was used to identify factors influencing prognosis. Receiver operating characteristic (ROC) curves were plotted to assess the predictive power of the number of involved regions for poor prognosis. Results:The inflammatory lesions of pancreas were divided into 13 regions: the lesser sac, pancreatic head and duodenum, left anterior renal, right anterior renal, left posterior renal, right posterior renal, left perirenal fat sac, right perirenal fat sac, left lateral abdominal wall, right lateral abdominal wall, left pelvic wall, right pelvic wall, and other regions. Significant differences were observed between the poor-prognosis group and the non-poor-prognosis group in terms of body mass index (BMI), pancreatic necrosis area, and the number of inflammatory lesion regions (all P<0.05). Multivariate logistic regression analysis showed that high BMI ( OR=1.723, 95% CI: 1.457-2.038, P<0.001), pancreatic necrosis area ≥50% ( OR=3.221, 95% CI: 1.073-9.668, P=0.037), and a higher number of inflammatory lesion regions ( OR=1.388, 95% CI: 1.110-1.735, P=0.004) were associated with a higher risk of poor prognosis in patients with ANP. Based on the number of inflammatory lesion regions, the ROC curve analysis revealed that the optimal cut-off value was 5.5 for predicting poor prognosis in patients with ANP, with an area under the curve of 0.747(95% CI: 0.680-0.815) and a sensitivity and specificity of 0.387 and 0.962, respectively. Conclusion:The peripancreatic membrane anatomy facilitates a relatively fixed partitioning of inflammatory lesions in patients with acute necrotizing pancreatitis, and the number of inflammatory lesion regions is associated with poor prognosis.

Objective:To investigate the status of hepatitis E virus infection in unpaid blood donors in Wenzhou.Methods:A total of 5 241 blood samples were selected from donors who successfully donated blood between October 2022 and March 2023. These samples were tested for hepatitis E virus (HEV) through HEV-IgG antibodies, HEV-IgM antibodies, and nucleic acid tests (HEV RNA). Samples that tested positive for HEV RNA underwent quantitative detection and sequencing. Data regarding the blood donors were collected, the positive rates of each test result were recorded, and the HEV infection status among different populations was analyzed.Results:The positive rate of the HEV-IgG antibody test was 23.60% (1 237/5 241), while the positive rate of the HEV-IgM antibody test was 0.29% (15/5 241). The positive rate of the HEV RNA test was 0.08% (4/5 241). The positive rate of the HEV-IgG test increased with age ( χ2 = 379.493, P < 0.05), and males had a higher positive rate than females ( χ2 = 5.12, P < 0.05). Both the positive rates of HEV-IgG and HEV-IgM tests increased as the number of blood donations increased ( χ2 = 40.87, 8.30, both P < 0.05). Furthermore, the positive rates were significantly higher in 2023 than in 2022 ( χ2 = 51.03, 13.35, both P < 0.05), and they were also significantly higher in spring than in winter ( χ2 = 51.03, 13.35, both P < 0.05). Only one HEV RNA-positive blood donor tested positive for both HEV-IgG and HEV-IgM antibodies, while the remaining three donors tested negative. Only one case was genotyped, and it belonged to genotype Ⅳ. Conclusion:The incidence of HEV infection among unpaid blood donors in Wenzhou of Zhejiang Province is relatively high, with the prevalent HEV genotype being type Ⅳ. Characteristics such as sex, age, and the number of blood donations can be utilized as auxiliary screening reference indicators.

Objective:To identify the independent factors of unplanned interruption during continuous renal replacement therapy (CRRT) and construct a risk prediction model, and to verify the clinical application effectiveness of the model.Methods:A retrospective study was conducted on critically ill adult patients who received CRRT treatment in the intensive care unit (ICU) of Zhejiang Hospital from January 2021 to August 2022 for model construction. According to whether unplanned weaning occurred, the patients were divided into two groups. The potential influencing factors of unplanned CRRT weaning in the two groups were compared. The independent influencing factors of unplanned CRRT weaning were screened by binary Logistic regression and a risk prediction model was constructed. The goodness of fit of the model was verified by a Hosmer-Lemeshow test and its predictive validity was evaluated by receiver operator characteristic curve (ROC curve). Then embed the risk prediction model into the hospital's ICU multifunctional electronic medical record system for severe illness, critically ill patients with CRRT admitted to the ICU of Zhejiang Hospital from November 2022 to October 2023 were prospectively analyzed to verify the model's clinical application effect.Results:① Model construction and internal validation: a total of 331 critically ill patients with CRRT were included to be retrospectively analyzed. Among them, there were 238 patients in planned interruption group and 93 patients in unplanned interruption group. Compared with the planned interruption group, the unplanned interruption group was shown as a lower proportion of males (80.6% vs. 91.6%) and a higher proportion of chronic diseases (60.2% vs. 41.6%), poor blood purification catheter function (31.2% vs. 6.3%), as a higher platelet count (PLT) before CRRT initiation [×10 9/L: 137 (101, 187) vs. 109 (74, 160)], lower level of blood flow rate [mL/min: 120 (120, 150) vs. 150 (140, 180)], higher proportion of using pre-dilution (37.6% vs. 23.5%), higher filtration fraction [23.0% (17.5%, 32.9%) vs. 19.1% (15.7%, 22.6%)], and frequency of blood pump stops [times: 19 (14, 21) vs. 9 (6, 13)], the differences of the above 8 factors between the two groups were statistically significant (all P < 0.05). Binary Logistic regression analysis showed that chronic diseases [odds ratio ( OR) = 3.063, 95% confidence interval (95% CI) was 1.200-7.819], blood purification catheter function ( OR = 4.429, 95% CI was 1.270-15.451), blood flow rate ( OR = 0.928, 95% CI was 0.900-0.957), and frequency of blood pump stops ( OR = 1.339, 95% CI was 1.231-1.457) were the independent factors for the unplanned interruption of CRRT (all P < 0.05). These 4 factors were used to construct a risk prediction model, and ROC curve analysis showed that the area under the curve (AUC) predicted by the model was 0.952 (95% CI was 0.930-0.973, P = 0.003 0), with a sensitivity of 88.2%, a specificity of 89.9%, and a maximum value of 1.781 for the Youden index. ② External validation: prospective inclusion of 110 patients, including 63 planned interruption group and 47 unplanned interruption group. ROC curve analysis showed that the AUC of the risk prediction model was 0.919 (95% CI was 0.870-0.969, P = 0.004 3), with a sensitivity of 91.5%, a specificity of 79.4%, and a maximum value of the Youden index of 1.709. Conclusion:The risk prediction model for unplanned interruption during CRRT has a high predictive efficiency, allowing for rapid and real-time identification of the high risk patients, thus providing references for preventative nursing.

Finding stable expression sites on the chromosomes of Chinese hamster ovary (CHO) cells is an effective method to solve the problem of unstable expression of CHO cells in long-term culture. Our group used lentiviral transfection to integrate the tracer gene (Zsgreen1) into the chromosome of CHO cells and found multiple potential stable expression sites. This study verified the ability of one of the sites located in the 148052-148157 bp region on chromosome NW_003614241.1 to stably express exogenous proteins.The expression of Zsgreen1 gene was first observed, and CRISPR/Cas9 technology was then used to integrate the enhanced green fluorescent protein (EGFP) gene into this site. Three strains of EGFP gene integrated cells were obtained. After 60 generations of suspension culture, the fluorescence intensity of the cells had no significant changes, which proved that this site can stably express the EGFP gene. The same method was used to construct recombinant CHO cell lines expressing the human serum albumin (HSA) gene, and was verified by Western blot that this site could express and secrete HSA. It shows that the above-mentioned sites can be integrated and can stably express exogenous proteins.

This study is performed to analyze the anti-liver fibrosis effect of the fusion protein of human serum albumin and extracellular domain of transforming growth factor beta type II receptor(eTGFBR2)in vivo to looking for the more stable anti-liver fibrosis drug. The mice model of liver fibrosis was constructed by CCl4 induction and the following groups are included in the study: the control group, CCl4 model group, the positive control group, eTGFBR2 treatment group, HSA-eTGFBR2 treatment group, and HSA group. Hematoxylin eosin staining, serum liver function index detection, and western blot are used to identify the anti-liver fibrosis activities. The results showed that: (1)CCl4 caused liver structure disorder, hepatocellular necrosis, collagen fibers proliferation, and induced liver fibrosis at last; (2)HSA-eTGFBR2 and its monomer drug improved the symptoms of liver fibrosis significantly, as well as reduced the damage of liver cells and collagen deposition, and recovered the liver basic structure to normal. Both of HSA-eTGFBR2 and its monomer drug improved liver function and reduced the expression level of liver fibrosis marker α-SMA and COL I. Moreover, the anti-liver fibrosis effect of the fusion protein is comparable to the monomer drug. In contrast, the albumin had no effect on therapeutic effect; (3)Reducing the injection frequency of HSA-eTGFBR2 achieved the comparable effects to the monomer drug with the normal injection frequency. In summary, the fusion protein HSA-eTGFBR2 has good anti-liver fibrosis effect. In addition, reducing the injection frequency of the fusion protein could also achieve the comparable treatment with the monomer drug, indicating that the fusion protein is stable and has longer half-lives and then a relatively positive application prospect in future.

Objective: Through a retrospective analysis of serology and nucleic acid screening data of unpaid blood donors in south Zhejiang region, the role of nucleic acid detection in the reduction of transfusion-related infectious diseases was discussed. Methods: 179 369 unpaid blood donation in south Zhejiong province from Jan, 2016 to Dec, 2016 was chosen. Enzyme linked immunosorbent (ELISA) test for blood index of infectious hepatitis B virus surface antigen (HBsAg), hepatitis C virus antibody (anti-HCV), human immunodeficiency virus antigen and antibody. At the same time the roche, haoyuan nucleic acid detection system were used for HBV-DNA, HCV-RNA, HIV-RNA in 6 or 8 doses mix samples three projects joint detection.NAT results of the serum with negative results in the serological tests were made statisticala analysis. Results: A total of 259 ELISA-/NAT+ samples were detected, HBV-DNA+ 255 cases, HCV-RNA+ 5 cases, one case of HBV-DNA and HCV-RNA + , with a positive rate of 0.14%. The analysis system of roche nucleic acid mix inspection rate of positive was 1.40%, and the split inspection rate of positive was 60.72%. The mix inspection positive rate of the haoyuan analysis system was 1.63%, and the split inspection rate of positive was 41.67%. Conclusions The detection of nucleic acid can make up the deficiency of serological test, and effectively reduce the leakage of transfusion-related infectious diseases, and ensuring the blood safety in this area.

This study was focus on investigating the anti-liver fibrosis effects of insulin-like growth factor-1 (IGF-1) in vitro.The effects of IGF-1 on human liver L-02 cell viability and cell cycle were observed.CC14-induced L-02 cell injury was set up to detect the anti-apoptotic activity of insulin-like growth factor-1 (IGF-1).Transforming growth factor β1 (TGF-β1) induced hepatic stellate cell line (HSC-T6) were used as a liver fibrosis model in vitro to analyze the effects of IGF-1 on the expression of liver fibrosis proteins and intracellular TGF-β1/Smad signaling pathway in HSC-T6 cells.The results showed that IGF-1 could relieve the growth inhibition effects of TGF-β1 on L-02 cells,increase the viability of L-02 cells injured by CCl4,decrease the expression of liver fibrosis proteins,and inhibit the TGF-β1/Smad signaling pathway by inhibiting the phosphorylation of Smad3.Our study suggested that IGF-1 exerted anti-liver fibrosis effects by stimulating L-02 cells proliferation,reducing cell damage and inhibiting ECM accumulation via interfering TGF-β1/Smad signaling pathway.

Aim To observe the physical coupling between transient receptor potential channel vanilloid type 4 (TRPV4 ) and cPLA2 in endothelial cells. Methods We investigated the physical association of TRPV4-cPLA2 coupling by immunofluorescence reso-nance energy transfer (immuno-FRET)to assess the spatial proximity between TRPV4 and cPLA2 in human microvascular endothelial cells (HMEC),primary cul-tured endothelial cells and in thoracic aortas rings from high salt-induced hypertension mice.Results At the cellular level,with high salt treatment,the physical in-teraction of TRPV4 and cPLA2 was significantly en-hanced in primary vascular endothelial cells and HMEC.Furthermore, in thoracic aortas rings from high salt-induced hypertension mice,we found an in-creases interaction between TRPV4 and cPLA2 in en-dothelial cells from arterial segments .Conclusion High-salt treatment increases the endothelial TRPV4-cPLA2 coupling,indicating that this coupling may pro-vide a new target for vascular endothelial dysfunction.

OBJECTIVE:To establish the HPLC fingerprint for Anemone raddeana. METHODS:HPLC was performed on the column of Phemomenex Gemini C18 with mobile phase of 0.1%phosphoric acid-acetonitrile(gradient elution)at a flow rate of 1 ml/min,the detection wavelength was 206 nm,the column temperature was 30℃,and the injection volume was 20μl. With the refer-ence of raddeanin A,13 batches of A. raddeana were analyzed,chromatographic fingerprint similarity evaluation system software was conducted for similarity analysis,and SPSS 13.0 was conducted for cluster analysis. RESULTS:There were 11 common peaks in the 13 batches of A. raddeana with similarity of higher than 0.90. According to the verification,the fingerprint and control fin-gerprint shows good consistency. The drugs in Huadian,Jiaohe, Tiangang,Shulan,Tonghua and Fusong of Jilin and Shangzhi of Heilongjiang were regarded as category 1,and in Harbin,Yabuli town and Yimianpo of Heilongjiang,Qingyuan of Liaoning,Ji-nan of Shandong were category 2. CONCLUSIONS:The established fingerprint can provide reference for the identification and quality evaluation of A. raddeana.