Developing and implementing biosafety standards for pathogenic microbiology laboratories is essential to achieving scientific, efficient, and standardized management and operation. This article analyzes the current standardization construction in biosafety in pathogenic microbiology laboratories domestically and internationally. It proposes a framework for the biosafety standard system of pathogenic microbiology laboratories, which mainly includes four parts: basic standards, management standards, technical standards, and industry applications. It provides a reference for the standardization work of pathogenic microbiology laboratories and helps to standardize the biosafety industry in China.

To create flexible and high-quality pediatric continuing education opportunities, the Children's Hospital of Zhejiang University School of Medicine has developed an online teaching system based on the medical consortium information system, mobile application, and website, which includes "5G+" digital classroom for case teaching, hierarchical diagnosis and treatment tracking, mobile courses, and live interactive lectures about difficult medical cases as support extensions of offline training. From 2020 to 2023, over 9 000 people participated in online activities. The results of questionnaire survey, theoretical examination, and semi-structured interview showed that the members of the medical consortium were satisfied with the training content and effect of the online medical continuing education model; medical personnel's theoretical levels were improved after training; the interviewees believed that online training was flexible and convenient with regard to time, location, and space, with positive impact for improving clinical ability and personal development. However, the online form cannot yet replace the traditional refresher training completely. In the future, we will establish a unified evaluation and assessment mechanism, form a standardized management system, and introduce advanced technologies, so as to encourage enthusiastic and effective participation, and promote the rational and efficient use of medical education resources.

Objective:To analyze the expression levels and clinical significance of interferon (IFN)-α/β in the renal cortex and serum of children with lupus nephritis (LN).Methods:A total of 32 children with LN diagnosed in the pediatric nephrology department of the Second Xiangya Hospital of Central South University from December 2017 to September 2020 were selected as the study subjects (LN group). The normal kidney control group consisted of 3 normal kidney transplant volunteers who underwent biopsy of kidney tissue (normal kidney control group), while 14 healthy children who underwent physical examination were collected as the normal control group. According to the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI), LN patients were divided into mild activity group ( n=8), moderate activity group ( n=9), and severe activity group ( n=15). According to the International Society of Nephrology/Society of Nephrology (ISN/RPS) 2003 LN classification criteria, pathological classification was performed (3 cases in the mild pathological damage group, 8 cases in the moderate pathological damage group, and 11 cases in the severe pathological damage group); Immunohistochemistry was used to detect the expression and distribution of IFN-α/β in glomeruli and renal interstitium; Enzyme linked immunosorbent assay (ELISA) was used to detect the concentration of IFN-α/β in serum samples and analyze its correlation with the pathological classification and disease activity of LN patients. Results:The serum and renal cortex IFN-α/β levels in the LN group were higher than those in the normal control group and normal kidney control group, respectively (all P<0.05). The average level of serum IFN-α/β in the heavy activity group was higher than that in the light and moderate activity groups (all P<0.05). The serum and renal cortex IFN-α/β levels in the severe pathological damage group were significantly higher than those in the mild and moderate pathological damage groups (all P<0.05). Conclusions:IFN-α/β in the renal cortex is closely related to renal injury in LN; Serum IFN-α/β can assist in evaluating the disease activity level of LN to a certain extent.

OBJECTIVE To provide reference for basic analysis of the pharmacodynamic substance in Stahlianthus involucratus. METHODS Overall 24 SD male rats were randomly divided into blank group （purified water）， and administration group （ethanol extract of S. involucratus， 15.75 g/kg， calculated by crude drug）， with 12 rats in each group. They were given drug liquid/purified water intragastrically， twice a day， every 6-8 h， for consecutive 3 days. After medication， the blood， urine and fecal samples were collected from two groups of rats. UPLC-Q-Exactive-MS technology was used to identify the chemical constituents in the ethanol extract of S. involucratus， and metabolites in the blood， urine and fecal of rats after intragastrical administration of the ethanol extract of S. involucratus. Multivariate statistical analysis was employed to screen various serum metabolites. Metabolic pathways were analyzed by MetaboAnalyst 5.0 platform. RESULTS A total of 38 chemical constituents were identified from the ethanol extract of S. involucratus， including fourteen prototype components and three metabolites identified from 5 urine samples， nine prototype components identified from fecal samples， and ten prototype components and one metabolite identified from serum samples. A total of 71 differential metabolites were screened from two groups of rat serum samples， of which 44 differential metabolites， such as ferulic acid， glycyrrhizin， were up-regulated and 27 differential metabolites， such as arachidonic acid， phenylacetylglutamine， were down-regulated. The 71 differential metabolites were mainly enriched in 11 metabolic pathways， including phenylalanine metabolism， linoleic acid metabolism， arachidonic acid metabolism， and tryptophan metabolism. CONCLUSIONS Ferulic acid， liquiritigenin， isofraxidin and formononetin may be the material basis that directly exert pharmacological effects of S. involucratus. S. involucratus may exert anti-inflammatory effects by affecting metabolic pathways， including arachidonic acid metabolism and tryptophan metabolism.

Objective To investigate the effects of dimethyl itaconate(DMI)on the secretion of pro-inflammatory factors in dendritic cells(DCs)and on the interphotoreceptor retinoid-binding protein(IRBP)1-20-specific T helper 17(Th17)cells in mice with experimental autoimmune uveitis(EAU).Methods Bilateral femur and tibia of C57BL/6J mice were isolated to obtain bone marrow cells,and these bone marrow cells were directionally induced with granulocyte macrophage-colony-stimulating factor(GM-CSF)and interleukin(IL)-4 to differentiate DCs.After 6 days,DCs were ran-domly divided into the DMI group and the phosphate-buffered saline(PBS)group.Cells in the DMI group were pretreated with 250 μmol·L-1 DMI,and cells in the PBS group were pretreated with the same volume of PBS for 3 hours.After-wards,100 μg·L-1 lipopolysaccharide was added in the every group to stimulate cells for 24 hours.The relative mRNA ex-pression levels of IL-6,IL-1 β,and IL-23 in DCs were measured by quantitative real-time polymerase chain reaction(qRT-PCR).The EAU model was constructed by actively immunizing mice with IRBP1-20,Freund's incomplete adjuvant,and my-cobacterium tuberculosis H37RA.Thirteen days after immunization,T cells in the spleen and lymph node isolated from EAU mice were cocultured with DMI-treatedor PBS-treated DCs in the medium containing IRBP1-20.They polarized toward Th17 cells.The percentage of Th17 cells in the cocultured cells was detected by flow cytometry.The IL-17 level in the coculture supernatant was detected by enzyme-linked immunosorbent assay(ELISA).qRT-PCR was performed to detect the relative mRNA expression levels of retinoid-related orphan receptor gamma t(RORγt),IL-17,IL-23R,and GM-CSF in the cocultured cells.Results qRT-PCR analysis revealed that the relative mRNA expression levels of IL-6,IL-1β,and IL-23 in the DMI group were significantly lower thanthose in the PBS group(all P＜0.05).Flow cytometry analysis showed that the proportion of Th17 cells in the cocultured cells in the DMI group was significantly lower than that in the PBS group(P＜0.05).ELISA analysis exhibited that the IL-17 level in the coculture supernatant in the DMI group was significantly lower than that in the PBS group(P＜0.05).The relative mRNA expression levels of IL-17,ROR-γt,IL-23R and GM-CSF in cocultured cells in the DMI group significantly decreased compared with the PBS group(all P＜0.05).Conclusion DMI can reduce the expression of IL-6,IL-1β and IL-23 in DCs,thus negatively modulating the responses of IRBP1-20-specific Th17 cells.

Objective:To investigate the effect of long noncoding RNA (lncRNA) 5033413D16Rik (lncRNA 5033413D16Rik) on the activity of interphotoreceptor retinoid-binding protein 1-20 (IRBP) 1-20-specific T helper 17 (Th17) cells in experimental autoimmune uveitis (EAU). Methods:Twelve SPF grade C57BL/6 female mice aged 8 to 10 weeks were selected and divided into EAU group and normal control group using the random number table method, with 6 mice in each group.Mice in the normal control group received no treatment.Mice in the EAU group were immunized with IRBP 1-20 emulsified in complete Freund's adjuvant to induce EAU.On day 13 after immunization, fundus examination and inflammation scoring were performed, and retinal histopathological changes were observed with hematoxylin and eosin staining.T cells were isolated from the spleen and lymph nodes of EAU mice, and the relative expression of lncRNA 5033413D16Rik was detected by real-time fluorescence quantitative PCR (qRT-PCR).In addition, the isolated T cells were divided into short hairpin RNA (shRNA)-5033413D16Rik transfected group and shRNA-negative control (NC) transfected group, and transfected with corresponding shRNAs, respectively.The knockdown efficiency of shRNA-5033413D16Rik was verified by qRT-PCR.T cells in the two groups were co-cultured with bone marrow-derived dendritic cells (BMDCs) under Th17 polarizing conditions.The relative expression of retinoic acid-related orphan receptor (RORγt), interleukin 17 (IL-17), IL-23 receptor (IL-23R), and granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA in co-cultured cells were analyzed by qRT-PCR.The IL-17 concentration in co-culture supernatants was assayed by ELISA and percentages of Th17 cells were determined by flow cytometry.The use and care of experimental animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals promulgated by the State Science and Technology Commission.The study protocol was reviewed and approved by the Experimental Animal Ethics Committee of Tianjin Medical University (No.TJYY2019110117). Results:EAU model was established successfully.qRT-PCR analysis showed that the expression of lncRNA 5033413D16Rik was significantly decreased in T cells from EAU mice compared with normal control mice ( t=-13.332, P<0.001).Compared with the shRNA-NC transfected group, the relative expression of lncRNA 5033413D16Rik in T cells of the shRNA-5033413D16Rik transfected group was significantly reduced ( t=-6.338, P<0.01).In co-cultured T cells, the expression levels of RORγt, IL-17, IL-23R, and GM-CSF mRNA in shRNA-5033413D16Rik transfected group were 1.61±0.13, 1.51±0.13, 1.85±0.33 and 1.45±0.11, which were significantly higher than 1.00±0.01, 1.00±0.01, 1.00±0.01 and 1.00±0.01 in shRNA-NC-transfected group ( t=-6.839, -8.221, -4.538, -4.189; all at P<0.05).ELISA analysis revealed that IL-17 concentraion in shRNA-5033413D16Rik transfected group was (2 350.39±367.02)pg/ml, which was significantly higher than (1 513.31±310.37)pg/ml ( t=-3.016, P=0.039).Flow cytometry showed that the percentage of Th17 cells in shRNA-5033413D16Rik transfected group was (17.20±0.44)%, which was higher than (14.10±0.84)% in normal control group ( t=-3.264, P=0.031). Conclusions:LncRNA 5033413D16Rik can inhibit the expression of pathogenicity related genes in antigen-specific Th17 cells and negatively regulates IRBP 1-20-specific Th17 cell responses.

The aim of this study is to investigate the immune effect of H9 subtype avian influenza virus(AIV)NP protein on mice and lay the foundation for the development of avian influenza vi-rus(AIV)vaccine.The H9N2 virus NP gene amplification product was cloned into the pET-32a expression vector,and the protein expression was verified by SDS-PAGE and Western blot,and the immune effect was evaluated by measuring the secretion of supernatant multicytokines in mouse splenocytes culture.The results showed that the total length of the coding region sequence of NP gene was 1 497 bp,NP recombinant proteins exist in both soluble and insoluble protein forms,and the specific bands were visible in Western blot.After immunizing mice,serum produces IgG-bind-ing antibodies with antibody titers of 1∶40 000.Compared with the control group,IL-2,IL-5 and IL-13 were significantly increased(P＜0.001),and the secretion of IL-6 was significantly increased compared with the control group.IL-4 and IL-12 p70 secretions were elevated compared with con-trols,but there was no significant difference.Compared with the control group,the secretions of IL-1β,IL-18,GM-CMF,TNF-α and IFN-γ were inhibited,but the difference was not significant(P＞0.05).The results showed that NP recombinant protein is a good immunogen,laying a foundation for in-depth research on influenza vaccine.

Diabetes mellitus(DM)is an endocrine metabolic disease mainly characterized by chronic hyperglycemia,which seriously threatens the health and quality of life of human beings,and with the improvement of living standard and unhealthy lifestyle in China,the incidence of DM continues to rise and tends to be younger,so it is urgent to carry out in-depth research on hypoglycemic treatment.DM is pathologically based on absolute or relative insulin deficiency,and there is no radical cure for it,and Western medicine mostly adopts insulin injection or oral hypoglycemic drugs for symptomatic treatment,which is effective but prone to toxic side effects in long-term use.Chinese medicine has the advantages of multi-path and multi-target in treating DM,and plays a role in lowering blood sugar by promoting insulin secretion,improving insulin resistance,regulating glucolipid metabolism and anti-oxidative stress,etc.Its efficacy is remarkable and the rate of toxic side effects is low.In recent years,there have been more studies on the mechanism of action of traditional Chinese medicine and compound prescriptions on animal models of DM.By reviewing the relevant literature in recent years,the author has systematically sorted out the mechanism of hypoglycemic action of single Chinese medicine,traditional Chinese medicine compound prescriptions,effective components of traditional Chinese medicine and their related experimental designs,from promoting insulin secretion,inhibiting gluconeogenesis,promoting glycogen synthesis,improving insulin resistance,inhibiting glycosidase activity,alleviating oxidative stress damage,inhibiting inflammatory response and regulating intestinal stress.The study and experimental design of the hypoglycemic mechanism of Chinese medicine were summarized in terms of promoting insulin secretion,inhibiting gluconeogenesis,promoting glycogen synthesis,improving insulin resistance,inhibiting glucosidase activity,alleviating oxidative stress damage,inhibiting inflammatory response and regulating intestinal flora,etc.,with a view to providing reference for the wider clinical application of Chinese medicine in hypoglycemia and its in-depth pharmacodynamic study.

Administrative multi-disciplinary team (MDT) is an efficient management approach that involves multiple administrative departments working together to solve cross functional systemic problems. Grid management improves management efficiency by constructing a three-level management network, efficiently transmitting information and coordinating resources. Starting from 2021, Children′s Hospital, Zhejiang University School of Medicine has adopted a combination of administrative MDT and grid management, with the average length of stay as the starting point to carry out medical efficiency management. Through the three stages of pre-, during-, and post-treatment, the hospital has reconstructed processes from technological innovation, information construction, team collaboration, and other aspects to shorten ineffective length of stay. After 3 years of practice, the average length of stay has been reduced from 6.41 d to 5.54 d, achieving an improvement in service efficiency while ensuring medical quality and safety.

This paper reviews the research progress on live-cell super-resolution fluorescence microscopy, discusses the current research status and hotspots in this field, and summarizes the technological application of super-resolution fluorescence microscopy for live-cell imaging. To date, this field has gained progress in numerous aspects. Specifically, the structured illumination microscopy, stimulated emission depletion microscopy, and the recently introduced minimal photon fluxes microscopy are the current research hotspots. According to the current progress in this field, future development trend is likely to be largely driven by artificial intelligence as well as advances in fluorescent probes and relevant labelling methods.
Artificial Intelligence ; Microscopy, Fluorescence ; Fluorescent Dyes ; Technology