Laboratory animals are the "living" tools of medical research. Through animal experiments, people can gain continuous insights into the laws of life, reveal the essence of diseases, develop vaccines and drugs for prevention and treatment, and play an important role in the technological development of fields related to human health. The environmental conditions of laboratory animals have a direct impact on their health, quality, and the results of animal experiments. The higher the degree of environmental control, the more reliable the experimental results are in terms of quality. Therefore, environmental control of laboratory animal facilities is important for ensuring that laboratory animals live under required conditions, which is a key factor for conducting effective animal experiments. This article analyzes the current status of environmental testing of laboratory animal facilities in Sichuan Province, briefly summarizing their number, area, and other basic information, and provides detailed statistics on the ability of institutions to conduct environmental testing for laboratory animal facilities in Sichuan Province. It also summarizes the testing requirements for laboratory animal facility environments based on national standards, regulatory requirements, and the quality control needs of facility users. In the analysis of testing indicators for laboratory animal facilities, based on testing data from 40 laboratory animal facilities in Sichuan Province, it was found that static pressure difference is the indicator most prone to non-compliance, followed by illumination and air exchange rate. Using barrier environments as examples, common problems in the process of environmental testing for laboratory animal facilities are summarized in six aspects: testing personnel, instruments, methods, technical materials, testing environment, and reports, and targeted suggestions are proposed. These suggestions help improve environmental control in laboratory animal facilities, and provide practical reference and guidance for relevant testing institutions, as well as laboratory animal producers and users in the industry.

ObjectiveTo provide reference for further strengthening the management of laboratory animals in Sichuan Province by reviewing and analyzing the results of parasitic and microbial quality inspections of laboratory animals from 2017 to 2023. Methods Sichuan Province has 31 licensed laboratory animal production units, with the main species including mice, rats, guinea pigs, rabbits, dogs, monkeys, and pigs. Sampling inspections and reports were conducted for units with laboratory animal production qualifications in accordance with current national and local standards for laboratory animal classification. The quality inspection results for various laboratory animals in Sichuan Province from 2017 to 2023 were analyzed. Results With the exception of 2018, annual quality inspections of laboratory animals were conducted every year between 2017 and 2023. Mice: In 2017, positive results for helminths, flagellates, Salmonella, murine hepatitis virus antibodies, and murine pneumonia virus antibodies were detected, with a pass rate of only 42.9%. In 2019, Staphylococcus aureus and Klebsiella pneumoniae were detected, with a pass rate of 86.7%. In 2021, Sendai virus antibodies were detected, yielding a pass rate of 85.7%.The pass rate in 2020, 2022, and 2023 was 100%. Rats: In 2017, positive results were found for helminths, mycoplasma antibodies, Staphylococcus aureus, Sendai virus antibodies, murine pneumonia virus antibodies, rat parvovirus RV strain antibodies, rat parvovirus (H-1) strain antibodies, and rat coronavirus antibodies, with a pass rate of 40.0%. In 2019, mycoplasma antibodies, Staphylococcus aureus and Klebsiella pneumoniae were detected, with a pass rate of 35.0%. No positive indicators were detected in 2020. In 2021, Sendai virus antibodies and rat parvovirus RV strain antibodies were detected, with a pass rate of 50.0%. In 2022, positive results for rat parvovirus RV strain antibodies were found, yielding a pass rate of 87.5%. In 2023, Pasteurella pneumotropica and Klebsiella pneumoniae were detected, with a pass rate of 85.7%. Dogs: In 2017 and 2019, the antibody titers for rabies virus and canine distemper virus were below the required standard, with pass rates of 33.3% and 20.0%, respectively. In 2020 and 2022, the pass rate was 100%. Guinea pigs (general grade): In 2019, positive results for Toxoplasma antibodies were detected, with a pass rate of 80.0%. In all other years, the pass rate was 100%. Monkeys: In 2019, positive results for Toxoplasma gondii and rhesus herpesvirus type I antibodies were found, with a pass rate of 87.5%.In 2020 and 2022, rhesus herpesvirus type I antibodies were positively detected, yielding pass rates of 93.3% and 97.5%, respectively. The pass rates in 2021 and 2023 were 100%. Clean guinea pigs, rabbits and pigs all passed the inspection each year. ConclusionIssues related to the parasitic and microbial quality of laboratory animals persist in Sichuan Province. Supervision and sampling inspections have proven to be effective in identifying these issues promptly, serving as a critical measure to ensure the quality of laboratory animals. The results of these inspections offer valuable data to support the healthy development of the laboratory animal industry in Sichuan Province.

Objective:To establish a model C-GALAD for detecting hepatocellular carcinoma (HCC) from the chronic liver disease and healthy people based on the serum markers.Methods:A clinical cohort including 229 hepatocellular carcinoma patients, 2 317 patients with chronic liver disease and 982 healthy people, was retrospectively collected from eight hospitals or physical examination institutions from April 2018 to October 2020. The data were divided into a training set and a testing set by stratified sampling with a 6∶4 ratio. A predictive model was established on the training set using a logistic backward regression method and validated on the testing set. In addition, clinical data from March to July 2021 in Beijing You′ an Hospital affiliated to Capital Medical University, including 84 patients with liver cancer and 204 patients with chronic liver disease collected were used for external independent validation of the model. The receiver operating characteristic curve (ROC) area under curve (AUC), the sensitivity and the specificity were used to evaluate the effectiveness of the model.Results:Through the logistic backward regression method, the seven signatures including age, gender, alpha-fetoprotein (AFP), alpha-fetoprotein alloplasm-3 ratio (AFP-L3%), des-gamma-carboxyprothrombin(DCP), platelet (PLT) and total bilirubin (TBIL) were selected as risk factors in the detection model. The area under the ROC curve (AUC) of the model on the testing set was 0.954, with an 88.04% sensitivity and a 94.85% specificity, and the AUC of model on the external independent validation set was 0.943, with an 89.29% sensitivity and a 90.2% specificity, which were better than other published models.Conclusion:The C-GALAD Ⅱ model can accurately predict the risk of hepatocellular carcinoma occurrence, and thus provide a trustworthy diagnosis method of hepatocellular carcinoma.

Polyhydroxyalkanoates (PHAs) are polymers obtained by esterification of hydroxy fatty acid monomers. Due to similar mechanical characteristics of traditional petroleum-based plastics, 100% biodegradability and biocompatibility, PHAs are considered to be one of the most potential green materials. However, the application and promotion of PHAs as a green and environmentally friendly material are difficult because of the high production costs. This article focuses on the current methods to reduce production cost of PHAs effectively, such as cell morphology regulation, metabolic pathway construction, economic carbon source utilization and open fermentation technology development. Despite most research results are still limited in laboratory, the research methods and directions provide theoretical guidance for the industrial production of economic PHAs.
Fermentation ; Industry ; Petroleum ; Plastics ; Polyhydroxyalkanoates

The biocompatibility of nanomaterials has attracted much attention. Graphene oxide (GO) is a nanomaterial widely used in biomedicine, but its toxicity can not be ignored. In this study, the effect of GO on the blood system (the hemolysis rate, the fragility of erythrocyte, and acetylcholinesterase activity) was systematically investigated. The results showed that the hemolysis rate of erythrocytes was lower than 8% when the GO concentration was below 100 μg/mL (P<0.01). GO at low concentration levels (<5 μg/mL) had no significant effect on the fragility of erythrocytes, but GO at high concentration (10 μg/mL) increased the fragility of erythrocytes (P=0.01). Moreover, GO increased the activity of acetylcholinesterase on erythrocytes. The concentration of 20 μg/mL graphene oxide with the size >5 μm (LGO) increased the activity of acetylcholinesterase by 42.67% (P<0.05). Then molecular dynamics simulation was used to study how GO interacted with acetylcholinesterase and increased its activity. The results showed that GO was attached to the cell membrane, thus may provide an electronegative environment that helps the hydrolysate to detach from the active sites more quickly so as to enhance the activity of acetylcholinesterase.
Acetylcholinesterase ; Erythrocytes ; Graphite ; Nanostructures

Objective:To evaluate perioperative coagulatory parameters and transfusion rates of lung transplantation recipients.Methods:Clinical data were retrospectively reviewed for 178 lung transplant recipients at China-Japan Friendship Hospital from March 2017 to July 2019. According to whether extracorporeal membrane oxygenation(ECMO)was used during perioperative period, they were divided into two groups of ECMO(131 cases)and without ECMO(47 cases). Clinical data, laboratory examinations and blood transfusion status of two groups were compared. In ECMO group, excluding secondary thoracotomy for hemostasis(7 cases)and incomplete data(2 cases), the remainders were divided into the groups of no red blood cell transfusion(63 cases), red blood cell transfusion(59 cases), plasma transfusion <1 000 ml(99 cases)and plasma transfusion≥1 000 ml (23 cases), no platelet transfusion(93 cases)and platelet transfusion(29 cases). Clinical data, laboratory examinations and ECMO-related parameters of recipients were analyzed by Bary Logistic regression.Results:Statistically significant inter-group differences existed in body mass index(BMI), disease course, primary disease, bilateral lung transplantation, laboratory examinations, postoperative blood transfusion volume, postoperative red blood cell and plasma transfusion ratio between groups with and without ECMO( P<0.05). Bilateral lung transplantation, ASA grade, differences in BMI, disease course, postoperative hemoglobin<100 g/L, postoperative PT/APTT/INR abnormalities and postoperative PLT count <100×10 9/L were independent risk factors for postoperative transfusion during ECMO. Conclusions:The application of ECMO during lung transplantation may affect the perioperative transfusion volume and demand.Fully assessing blood transfusion requirements, optimizing coagulation monitoring and identifying the independent influencing factors of postoperative blood transfusion facilitate clinical scientific and rational blood transfusions.

【Objective】 To study the effect of intravenous immunoglobulin(IVIG) on the detection of blood transfusion compatibility in patients. 【Methods】 56 patients, submitted to our Hospital from March 1, 2017 to December 31, 2020, were enrolled as the research objects. They had negative unexpected antibody screening, major crossmatch incompatibility with the same blood type donors, and had a history of IVIG infusion. ABO and RhD blood groups typing, unexpected antibodies screening, crossmatch, direct antiglobulin test, indirect antiglobulin test, and acid elution test were all conducted by microcolumn gel method. 【Results】 After IVIG infusion, the initially major crossmatch incompatibility with the same blood type donors turned into compatiblity with O-type donors. Among them, 2 patients had transient discrepancy in ABO forward and reverse blood typing due to the IVIG infusion. IgG anti-A were detected in the red blood cell elution of 37 A-type patients; IgG anti-B in 2 B-type patients; 3 cases of IgG anti-A+ anti-B and 14 cases of solo IgG anti-A in 17 AB-type patients. 3 batches of IVIG preparations were detected randomly, IgG anti-A titer was 32-64, and IgG anti-B titer was 8-16. 【Conclusion】 The discrepancy in ABO forward and reverse blood typing and major crossmatch incompatibility with the same blood type donors may occur after non-O type patients received IVIG, which contains IgG types of anti-A and anti-B. In this situation, it is recommended to prepare major crossmatched O-type washed red blood cells to ensure the safety and effectiveness of clinical blood transfusion.

【Objective】 To explore the influencing factors of perioperative red blood cell transfusion in patients underwent lung transplantation, so as to provide reference for perioperative blood management (PBM) of lung transplantation patients. 【Methods】 The clinical data of 173 lung transplant patients completed in China-Japan Friendship Hospital from March 2017 to June 2019 were retrospectively analyzed. The patients were divided into two groups according to perioperative red blood cell transfusion volume: large blood transfusion group (transfusion red blood cell volume ≥6 U, n=66) and non-large blood transfusion group (red blood cell transfusion volume <6 U, n=107). The basic information, preoperative laboratory test results, and surgical status of the two groups were statistically analyzed.The clinical data of the two groups were analyzed by univariate analysis. The factors of P<0.15 were included in the binary logistic regression analysis, and the independent influencing factors of perioperative massive blood transfusion in patients with lung transplantation were found. 【Results】 Univariate analysis of clinical data of the two groups of patients (large blood transfusion group vs. non-large blood transfusion group) showed that the differences of smoking history ratio [44(66.7%) vs 87(81.3%)], BMI(20.8±4.5 vs 22.5±4.0)(P<0.05), preoperative Hb [124(111, 138.8) vs 138(126, 149)], preoperative Hct [37.9(34.8, 42.5) vs 41.3(37.9, 44.6)], surgery duration(327.9±107.7 vs 238.4±77.0), intraoperative blood loss(1 108.6±1342.0 vs 341.8±270.8) and single lung transplantation [28(42.4%) vs 84(78.5%)] (P<0.01) were statistically significant. Logistic regression analysis showed that intraoperative blood loss (OR=1.001, P<0.05), surgery duration (OR=1.006, P<0.05), preoperative Hb (OR=0.973, P<0.01), lung transplantation type(single or double lung transplantation)( OR=0.247, P<0.05) and extracorporeal membrane oxygenation (ECMO) (OR=0.187, P<0.01) were independent factors influencing red blood cell transfusion during lung transplantation. 【Conclusion】 Intraoperative blood loss and surgery duration are risk factors for massive blood transfusion during the perioperative period. And the use of ECMO, preoperative Hb, single lung transplantation (compared to double lung transplantation) are protective factors for perioperative massive blood transfusion.

Objective To study the construction of tissue engineering fat filler and the survival of fat particles.Methods Stromal vascular fraction (SVF) was derived from inguinal subcutaneous fat of rabbits,mixed with autologus mature fat particles and with or without neuropeptide Y to construct PLGA composite tissue.The complex was subcutaneously transplantef into back sites of the rabbits.Based on the different combination five groups were divided:Group A:PLGA + mature fat particles without NPY;Group B:PLGA + mature fat particles + NPY;Group C:PLGA + complex of SVF with mature fat particles without NPY;Group D:PLGA + complex of SVF with mature fat particles + NPY;Group E:Complex of SVF with mature fat particles + small ball with NPY.Diffenrence of virous constructive ways and fat particle survival was evaluated by general observation,histological staining,fluorescence tracing at two weeks,one month and three month after operation.Results Group D was superior to groups A,B,C and E in survival volume,graft texture and vascularization at one mouth and three mouths.The fat average srvival rate was 57.5±2.5％.Fat cell grew well,and precursor cells proliferated and differentiated actively.Conclusions High quality tissue engineering materials are successfully established with SVF-mature fat particle complex,PLGA and NPY,which could obviously improve fat particle transplantation.

Objective:To investigate the effect of IL-17A on the differentiation and maturation of murine bone marrow-derived dendritic cells( BMDCs ) . Methods: Murine bone marrow cells were isolated and cultured in RPMI1640 complete medium in the presence of GM-CSF(20 ng/ml) for 8 days to induce differentiation of murine bone marrow cells to DC progenitors. Then these cells were treated with LPS(1 μg/ml) for 36 h which polarized immature DCs into mature DCs. Different concentrations of rmIL-17A(10 or 100 ng/ml) was added to the culture medium at different stages of BMDC differentiation and maturation. Co-stimulatory molecules expression on BMDC were analyzed by flow cytometry,and the culture supernatants were analyzed for IL-12p40 and IL-10 level by ELISA. Results:rmIL-17 could promote co-stimulatory molecules( CD40,CD80,CD86 and MHCⅡ) expression on BMDCs in a does-dependent manner,especially,the expression of CD40 and MHCⅡhad a significant increase in high concentration of rmIL-17A group;rmIL-17A was added while LPS induced maturation of BMDCs. CD40,CD80,CD86 and MHCⅡexpression on BMDC increased sharply in LPS plus rmIL-17A stimulation group,besides,CD86,MHCⅡ showed a higher level expression on BMDC with the increase of con-centration of rmIL-17A. Furthermore,secretion of IL-12p40 and IL-10 increased significantly in the group of DCs treated with LPS plus low concentration of rmIL-17 compared with the group without rmIL-17(P<0. 001). However,high concentration of rmIL-17A group showed significantly higher levels of IL-12p40(P<0. 001),but there was no difference in IL-10. Conclusion:IL-17A promotes the phe-notypic development of BMDC progenitors propagated in GM-CSF and cooperate with LPS to induce BMDC differentiation and matura-tion.