Objective To observe the protective effects of FTY720 on the Con A-induced mouse hepatic fibrosis injury,and to find the possible mechanisms of protective effects.Methods The pathologic models of hepatic fibrosis injury in the mice caused by Con A were set up.Forty mice were randomly divided into control group, model group,high dose of FTY720 (4 mg·kg-1 )group and low dose of FTY720 (1 mg·kg-1 )dose group (n=10).The serum alanine aminotransferase (ALT)and asparate aminotransferase (AST)activities,hepatic index and pathological changes of hepatic tissue were detected .Results Compared with model group,the serum ALT and AST activities in low and high doses of FTY720 groups were decreased significantly (P < 0.05 or P < 0.01).The optical microscope results showed that there were inflammatory cells and hepatocellular necrosis in model group. The masson staining results showed that there were surrounding fiber bundle and hepatic lobule fusion in model group;compared with model group,the damage degree in low and high doses of FTY720 groups was reduced.The protective effects of FTY720 on hepatic injury showed linear relation to the drug dose.Conclusion FTY720 could decrease the levels of ALT/AST,thus FTY720 alleviate hepatic damage degree and delay the process of hepatic fibrosis.The protective effects of FTY720 on hepatic injury in experimental hepatic fibrosis mice may be related to the mechanisms mentioned above.

Objective To study the damage effect of oxidized low density lipoprotein (OX-LDL)on the endothelial cells and its influence in the expressions of Toll like receptor 4 (TLR-4)and epithelial cell adhesion molecule (EpCAM).Methods The human endothelial cells ECV-304 were cultured in vitro and treated by different concentrations of OX-LDL for 24 h,and the cell vitality was measured by Taipan blue rejection test.The reactive oxygen species (ROS)level,mitochondrial membrane potential (MMP),cell cycle and apoptosis were detected by flow cytometry (FCM).The expression of TLR-4 and EpCAM were also analyzed by FCM.Results Compared with control group,the ROS levels in ECV-304 cells in 25,50,100,and 200 mg·L-1 OX-LDL were increased, but the cell vitalities and MMP were decreased (P < 0.05).Compared with control group,the percentages of S phase cells in 25 and 50 mg·L-1 OX-LDL groups were increased,the G0 G1 phase cells were slightly reduced,and the SubG1 phase cells (apoptotic cells)were increased with the increasing of OX-LDL concentration.Compared with control group,the TLR-4 and EpCAM expression levels in ECV-304 cells in 50 mg·L-1 OX-LDL group at 24 h were significantly increased (P < 0.05 ). Conclusion OX-LDL could increase the ROS level and induce the apoptosis in vascular endothelial cells,while increase the cell vitality and MMP.During the damage process,the expresion levels of TLR-4 and EpCAM are up-regulated.

Fresh healthy human erythrocytes were treated by four kinds of bile acid solutions (DC, GDC, C, and GC) at physiological and pathological concentrations of human plasma, and the membrane proteins, enzyme activities and permeability of cations were observed. The results showed that some components of the membrane cytoskeleton proteins disappeared, the activities of total ATPase and Gr-ATPase were distinctly increased with the increasing of bile acid concentrations, the activity of the membrane acetylcholinesterase was decreased with the increasing bile acid concentrations, the permeability of cations was increased with the increasing bile acid concentrations. The effect of the dihydroxy bile acids more stronger than that of the trihydroxy bile acids.

Objective To study the apoptosis of K562 cells induced by a new immunosuppressive agent FTY720 and its mechanism,and to provide experimental basis for the treatment of leukemia in clinic.Methods The K562 cells were cultured in vitro and divided into blank control group and FTY720 treatment group.The K562 cells in FTY720 treatment group were treated with 6μmol·L-1 FTY720 for 3,6,and 12 h,or treated with different concentrations of FTY720 (2,4,6,8μmol·L-1)for 24 h.The apoptosis,level of reactive oxygen species (ROS),mitochondrial membrane potential(MMP)and cell cycle were measured by flow cytometry.The inhibitory rate of proliferation of K562 cells after treated with FTY720 was detected by WST-1 reducting assay.Results The results of flow cytometry showed that the percentages of apoptotic cells were increased after treated with 6μmol·L-1 FTY720 for 3,6,and 12 h with the prolongation of time compared with blank control group(P<0.01).The percentages of apoptotic cells were also increased after treated with different concentrations of FTY720 for 24 h compared with blank control group(P<0.01).Compared with blank control group,the ROS levels were increased with the increasing of FTY720 concentration,while the MMP was decreased(P<0.01).Compared with blank control group,the percentage of cells at G0/G1 phase was increased,while those at S and G2/M phases were decreased with the increasing of FTY720 concentration (P<0.05).The WST-1 reduction assay results indicated that the inhibitory rates of proliferation of K562 cells after treated with 2,4,6,and 8μmol·L-1 FTY720 for 72 h were (24.0±4.1)%,(46.4±3.9)%,(67.0±4.8)%,and (88.2±5.6)%,respectively,compared with blank control group.The concentration of FTY720 to result the inhibitory rate of 50% (IC50 )on K562 cells was 5.5μmol·L-1 .Conclusion FTY720 could inhibit the proliferation of K562 cells by blocking cell cycle and inducing apoptosis through provoking ROS.

Compared with the widely used rodents,pigs are anatomically,physiologically,and genetically more similar to humans,making them high-quality models for the study of liver diseases.Here,we review the latest research progress on pigs as a model of human liver disease,including methods for establishing them and their advantages in studying cystic fibrosis liver disease,acute liver failure,liver regeneration,non-alcoholic fatty liver disease,liver tumors,and xenotransplantation.We also emphasize the impor-tance of genetic engineering techniques,mainly the CRISPR/Cas9 system,which has greatly enhanced the utility of porcine models as a tool for substantially advancing liver disease research.Genetic engineering is expected to propel the pig as one of the irreplaceable animal models for future biomedical research.

Andrographis paniculata is a traditional Chinese herbal medicine with a wide cultivation in south China. The market demand has been increasing steadily in recent years. However, there are problems such as disordered production, pesticide residues and heavy metal content in the process of Andrographis paniculata production. Pollution- free cultivation is an effective strategy to solve this problem, and it is also the development direction of the Chinese herbal medicine planting industry at this stage. In order to guide the cultivation of pollution-free and high-quality Andrographis paniculata, this study established a pollution- free fine cultivation technology system of Andrographis paniculata including precise cultivation and site selection. In addition, soil improvement, scientific pollution-free planting mode and high- efficiency integrated pest control technology help solve problems such as pesticide residues and heavy metal content. This paper provides a reference for the pollution-free and fine cultivation of Andrographis paniculata.

Objective To investigate the mechanism of action of Dingchuan granules in intervening respiratory syncytial virus pneumonia based on network pharmacology and animal experiments.Methods The potential active ingredients and targets of each traditional Chinese medicine in Dingchuan granules were obtained from TCMSP and TCMID databases,and the active ingredients of the drugs in Dingchuan granules were screened according to ADME pharmacokinetic parameters;the potential disease targets of respiratory syncytial virus pneumonia were obtained from Genecards,OMIM,and DisGeNet databases;the protein-interaction networks of intersecting targets were visualized by using String platform;the key core targets were visualized by using David database;and the intersection targets were visualized by using David database.Interaction networks were constructed using the String platform to visualize the intersecting targets;GO functional enrichment and KEGG pathway enrichment analyses of the key core targets were performed using the David database;and then the relevant networks for the intervention of Respiratory Syncytial Virus Pneumonia in the Dingchuan particles were constructed using the Cytoscape software(3.9.1).Respiratory syncytial virus(RSV)-induced respiratory syncytial virus pneumonia in young rats was selected for experimental validation.Results The results of the network pharmacology showed that the 177 potential active ingredients of Dingchuan granules acted on 144 targets,and there were 1075 targets related to respiratory syncytial virus pneumonia,and 55 drug-disease intersecting targets were obtained,of which 25 core targets were ALB,IL6,CASP3,EGFR,VEGFA,etc.The GO function of Dingchuan granules was also investigated,and the GO function of Dingchuan was investigated.The results of GO function enrichment analysis showed that the biological processes involved mainly include positive regulation of transcription,positive regulation of RNA polymerase II promoter transcription,positive regulation of gene expression,negative regulation of apoptosis,etc.The KEGG pathway enrichment mainly involves the PI3K-Akt signaling pathway,the cancer pathway,the tumor necrosis factor signaling pathway,the IL-17 signaling pathway and so on.The KEGG pathway enrichment mainly involves PI3K-Akt signaling pathway,tumor necrosis factor signaling pathway,IL-17 signaling pathway,etc.Animal experiments initially showed that the fixed wheezing granules can play a role in inflammation and immune response by regulating the PI3K-Akt signaling pathway,thus participating in the inflammatory and immune response.Compared with the normal group,the ratios of p-PI3K/PI3K and p-Akt/Akt in the lung tissues of young rats in the model group were significantly higher(P<0.05),the viral load was significantly higher(P<0.05),the pathological score of lung tissue damage was significantly higher(P<0.05),and the content of inflammatory factors IL-6 and TNF-α in alveolar lavage fluid(BALF)was significantly higher(P<0.05).Compared with the model group,the expression of p-PI3K/PI3K and p-Akt/Akt proteins in the lung tissues of young rats in each dose group and the positive control group was reduced(P<0.05),the viral load was significantly reduced(P<0.05),the pathological scores of lung tissue injury were significantly reduced(P<0.05),and the contents of inflammatory factors IL-6 and TNF-α in BALF were significantly reduced(P<0.05).Conclusion The study revealed the synergistic mechanism of multi-component,multi-target,and multi-pathway action of Dingchuan granules for the treatment of respiratory syncytial virus pneumonia.It was verified by animal experiments that RSV infection in young rats probably activated the PI3K-Akt signaling pathway,which caused the release of inflammatory factors IL-6 and TNF-α.Dingchuan granules could effectively down-regulate the PI3K-Akt signaling pathway,inhibit the release of inflammatory factors IL-6 and TNF-α,and thus achieve the anti-inflammatory effect.

Objective:To explore the patterns and causes of occupational exposure to infectious diseases (OEID) among frontline medical staffs (FMS) in coronavirus disease 2019 (COVID-19) isolation wards (CIW), and the particularity of post-OEID management and the measures to prevent OEID.Methods:A total of 1 061 FMS of Wuhan Huoshenshan Hospital from February 4 to March 21, 2020 were enrolled. The OEID of FMS was investigated and analyzed from the perspectives of FMS physical and psychological conditions, protective equipment, infection-control related regulations and procedures, local air quality, exposure patterns, and the particularity of emergency treatment after exposure.Results:The incidence of OEID among FMS was 2.0%(21/1 061). The nurses and doctors accounted for 95.2%(20/21) and 4.8%(1/21), respectively. The incidences in 17 general wards and two intensive care units (ICU) were 71.4%(15/21) and 28.6%(6/21), respectively. Nearly 90.5%(19/21) and 9.5%(2/21) of the OEID events occurred in contaminated area and potential contaminated area, respectively. About 23.8%(5/21) of the OEID events were air exposure of oral-nasal skin, mucosa and respiratory tract, which was secondary to uncontrollable vomiting, and 76.2%(16/21) were pricking injuries. The inducement factors involved poor quality and inappropriate wearing of some goggles, atomization of the inside of goggles leading to blurring vision, chest distress and decreased sense of touch and operational flexibility related to level-3 protection equipment, poor air quality, FMS physical and psychological conditions, etc. Under the direction of "the Procedures for Handling OEID" , all incidents are properly handled and no FMS was infected by 2019 novel coronavirus and blood-borne pathogens. No new OEID event was found after the strict implement of set of preventive measures.Conclusions:The OEID among FMS in CIW is attributed to multiple causes. The optimized process that takes into account the specificity of OEID management for both COVID-19 and blood-borne infectious diseases can effectively prevent potential post-exposure infections. And reasonable precautions can fully reduce the risk of OEID of FMS in CIW.

A 31-year-old man sought medical evaluation for a 2-year history of edema and proteinuria, with prior pathology suggesting atypical membranous nephropathy (MN). Despite treatment with a combination of steroids, calcineurin inhibitors, and four courses of rituximab (1 g, intravenous injection), the patient′s nephrotic syndrome showed no relief (24 h urine protein peaked at 31.18 g/d), indicating refractory nephrotic syndrome. Later in the disease course, a sudden surge of creatinine level (322.5 μmol/L) prompted a renal biopsy, which revealed concurrent acute interstitial nephritis. Further treatment involving steroids, cyclophosphamide, and a fifth rituximab infusion (1 g, intravenous injection) resulted in improvement in renal function (serum creatinine: 322.5?147 μmol/L), but the MN failed to achieve partial relief. Subsequent treatment with the novel humanized CD20 monoclonal antibody obinutuzumab (1 g, intravenous injection) was initiated. In the latest follow-up, anti-phospholipase-A2-receptor antibody (PLA2R) antibody were negative, B cells were eliminated, serum albumin was 36 g/L, urine protein-to-creatinine ratio was 4 810 mg/g, and serum creatinine was 162 μmol/L. This case underscores the potential efficacy of obinutuzumab in refractory MN. For advanced MN cases, prompt identification of the cause of acute kidney injury is crucial, emphasizing the need for targeted interventions to potentially stall renal function decline.

ObjectiveTo explore the potential mechanism of Dingchuan Granule （定喘颗粒， DG） in the treatment of respiratory syncytial virus （RSV） pneumonia. MethodsA total of 60 male Sprague Dawley （SD） rats were randomly divided into control group， model group， ribavirin group， DG low-dose group， DG middle-dose group， and DG high-dose group， with 10 rats in each group. Except for the control group， rats were administrated with RSV via intranasal drip. After model establishment， the DG low-， middle-， and high-dose groups were administrated via oral gavage with DG at 3.47， 6.93， and 13.86 g/（kg·d） respectively， while the ribavirin group was administrated via oral gavage with ribavirin at 15.75 mg/（kg·d）. The drug was given once daily for one week. The rats in the control group and the model group were not given any drug， only subjected to the grasping action. Twenty-four hours after the last administration， the pathological changes of lung tissues were observed and scored using HE staining. The levels of serum inflammatory factors， including tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6）， were detected by colorimetry. The protein levels of nuclear factor （erythroid derived 2）-like 2 （Nrf2）， Kelch-like ECH-associated protein 1 （Keap1）， heme oxygenase 1 （HO-1）， and NAD（P）H quinone dehydrogenase 1 （NQO1） in lung tissues were measured by Western Blot. The RSV load as well as the gene expression levels of Nrf2， Keap1， HO-1， and NQO1 in lung tissues were determined by qRT-PCR. The level of reactive oxygen species （ROS） in rat lung tissues was detected using chemiluminescence. The levels of glutathione （GSH） and malondialdehyde （MDA） in rat lung tissues were measured by a microassay. ResultsCompared with the control group， other groups had significant increases in pathological score of lung tissue， RSV load， levels of ROS， MDA， serum TNF-α， IL-1β， and IL-6； decrease in GSH level， increases in expression level of Keap1 protein and its mRNA in lung tissue， and significant decrease in levels of Nrf2， HO-1， expression level of NQO1 protein and its mRNA （P＜0.05）. Compared with the model group， all the above-mentioned indicators in the DG low-， middle-， and high-dose groups and the ribavirin group were improved to varying degree （P＜0.05）. The levels of serum TNF-α， IL-1β， and IL-6 in rats of DG dose groups showed a dose-dependent pattern， the DG high-dose group exhibiting the best effect （P＜0.05）. The DG high-dose group was superior to the DG low- and middle-dose groups in reducing the levels of ROS and MDA， and increasing the level of GSH in lung tissues （P＜0.05）. The DG high-dose group and the ribavirin group had better effect than the DG middle-dose group in reducing the RSV load （P＜0.05）. The DG high-dose group was superior to the ribavirin group in improving the protein levels of Nrf2， Keap1， HO-1， and NQO1 （P＜0.05）. ConclusionDG could inhibit oxidative stress by regulating the Nrf2/Keap1/HO-1/NQO1 signaling pathway to improve pulmonary inflammation and treat RSV pneumonia， with the DG high-dose group showing the best effect.