Developing and identifying effective medications and targets for treating hepatic fibrosis is an urgent priority. Our previous research demonstrated the efficacy of artesunate (ART) in alleviating liver fibrosis by eliminating activated hepatic stellate cells (HSCs). However, the underlying mechanism remains unclear despite these findings. Notably, endocytic adaptor protein (NUMB) has significant implications for treating hepatic diseases, but current research primarily focuses on liver regeneration and hepatocellular carcinoma. The precise function of NUMB in liver fibrosis, particularly its ability to regulate HSCs, requires further investigation. This study aims to elucidate the role of NUMB in the anti-hepatic fibrosis action of ART in HSCs. We observed that the expression level of NUMB significantly decreased in activated HSCs compared to quiescent HSCs, exhibiting a negative correlation with the progression of liver fibrosis. Additionally, ART induced senescence in activated HSCs through the NUMB/P53 tumor suppressor (P53) axis. We identified NUMB as a crucial regulator of senescence in activated HSCs and as a mediator of ART in determining cell fate. This research examines the specific target of ART in eliminating activated HSCs, providing both theoretical and experimental evidence for the treatment of liver fibrosis.
Hepatic Stellate Cells/cytology* ; Liver Cirrhosis/genetics* ; Artesunate/pharmacology* ; Cellular Senescence/drug effects* ; Membrane Proteins/genetics* ; Animals ; Humans ; Nerve Tissue Proteins/genetics* ; Tumor Suppressor Protein p53/genetics* ; Male ; Mice

As main recipient cells for porcine reproductive and respiratory syndrome virus (PRRSV), porcine alveolar macrophage (PAM) are involved in the progress of several highly pathogenic virus infections. However, due to the fact that the PAM cells can only be obtained from primary tissues, research on PAM-based virus-host interactions remains challenging. The improvement of induced pluripotent stem cells (iPSCs) technology provides a new strategy to develop IPSCs-derived PAM cells. Since the CD163 is a macrophage-specific marker and a validated receptor essential for PRRSV infection, generation of stable porcine induced pluripotent stem cells lines containing CD163 reporter system play important roles in the investigation of IPSCs-PAM transition and PAM-based virus-host interaction. Based on the CRISPR/Cas9- mediated gene editing system, we designed a sgRNA targeting CD163 locus and constructed the corresponding donor vectors. To test whether this reporter system has the expected function, the reporter system was introduced into primary PAM cells to detect the expression of RFP. To validate the low effect on stem cell pluripotency, we generated porcine iPSC lines containing CD163 reporter and assessed the pluripotency through multiple assays such as alkaline phosphatase staining, immunofluorescent staining, and EdU staining. The red-fluorescent protein (RFP) expression was detected in CD163-edited PAM cells, suggesting that our reporter system indeed has the ability to reflect the expression of gene CD163. Compared with wild-type (WT) iPSCs, the CD163 reporter-iPSCs display similar pluripotency-associated transcription factors expression. Besides, cells with the reporter system showed consistent cell morphology and proliferation ability as compared to WT iPSCs, indicating that the edited-cells have no effect on stem cell pluripotency. In conclusion, we generated porcine iPSCs that contain a CD163 reporter system. Our results demonstrated that this reporter system was functional and safe. This study provides a platform to investigate the iPS-PAM development and virus-host interaction in PAM cells.
Swine ; Animals ; Induced Pluripotent Stem Cells/metabolism* ; Receptors, Cell Surface/genetics* ; Antigens, CD/metabolism* ; Porcine respiratory and reproductive syndrome virus/genetics*

Small cell lung cancer （SCLC） accounts for about 15% in lung cancer and is highly malignant， heterogeneous and invasive. Etoposide combined with platinum-based chemotherapy is the basis of standard first-line treatment for extensive-stage SCLC， but suffers from the problem of susceptibility to drug resistance and relapse. In recent years， the emergence of new immunological drugs and novel cytotoxic drugs has improved the survival of SCLC patients to a certain extent， especially bringing therapeutic hope to patients with relapsed/refractory SCLC. In this paper， we review the current clinical drug regimens and the new progress of potential target drug therapeutic regimens for the treatment of SCLC. At present， the first-， second- and third-line schemes of SCLC include etoposide+carboplatin， atezolizumab+etoposide+platinum， adebrelimab， topotecan， docetaxel， etc.； the current drug targets for the treatment of SCLC mainly focus on topoisomerase Ⅱ/Ⅰ， DNA， the immune checkpoint molecules programmed death-1/programmed death-ligand 1， tubulin， etc. The potential target drug therapeutic options include alisertib+ paclitaxel， rovalpituzumab， APG-1252， etc.， and mainly focus on DNA damage response pathways and immune pathways， which can achieve the prolongation of patient survival by exerting anti-tumor effects through aurora kinase A and other potential targets.

Objective:To explore the correlation of miRNA-509 (miR-509) expression level with the clinical characteristics and prognosis in acute myeloid leukemia (AML) patients.Methods:The RNA-seq expression data and clinical data of 162 newly diagnosed AML patients (non-M 3 subtype) based on the World Health Organization (WHO) classification were obtained from the Cancer Genome Atlas (TCGA) database (Project ID:TCGA-LAML). The final follow-up time was April 30th, 2013. According to the different treatment methods, all cases were firstly divided into the chemotherapy group and allogeneic hematopoietic stem cell transplantation (allo-HSCT) group. Each group was subdivided into high miR-509 group and low miR-509 group according to the median relative expression of miR-509, respectively; the clinical characteristics of both groups were analyzed. The effects of miR-509 relative expression level on overall survival (OS) and event-free survival (EFS) of AML patients in chemotherapy group and allo-HSCT group were compared. On the other hand, cases were divided into two groups based on the median relative expression level of miR-509, then each group were further divided into the chemotherapy subgroup and allo-HSCT subgroup according to different treatment methods. The differences of OS and EFS of AML patients with different miR-509 expression in chemotherapy and allo-HSCT subgroups were compared. Results:All the 162 AML patients were firstly divided into chemotherapy group (90 cases) and allo-HSCT group (72 cases). Each group was subdivided into high miR-509 and low miR-509 group according to the median relative expression of miR-509 (chemotherapy group: 14.07, allo-HSCT group: 14.85), respectively. There were no statistically significant differences in the proportion of patients with gender, white blood cell count at initial diagnosis, bone marrow blast/naive cells ratio, peripheral blood blast/naive cells ratio and France-America-British (FAB) subtype between high miR-509 and low miR-509 subgroups in chemotherapy subgroup and allo-HSCT subgroup (all P > 0.05). In the chemotherapy group, low miR-509 group comprised more cases with intermediate prognosis compared to high miR-509 group[68.9% (31/45) vs. 42.2% (19/45), χ2 = 6.48, P = 0.011]. No significant differences in the proportion of patients with other risk stratification were found between both subgroups (all P > 0.05). Of the 90 cases in chemotherapy group, the median OS time in low miR-509 group (45 cases) and high miR-509 group (45 cases) was 10.2 months and 6.7 months, respectively. The 5-year OS rates of the two subgroups in chemotherapy group were 17.9% and 17.0%, and the 5-year EFS rates were 16.9% and 18.2%.There were no significant differences in OS and EFS of low miR-509 group and high miR-509 group ( P = 0.575, P = 0.436). In the allo-HSCT group (72 cases), longer OS and EFS were observed in low miR-509 group compared with high miR-509 group, and the differences were statistically significant ( P = 0.006, P = 0.022). All the 162 cases were divided into low miR-509 group (81 cases) and high miR-509 group (81 cases) based on the median expression level of miR-509 (14.19). In the low miR-509 expression group, cases administered allo-HSCT had a better OS in comparison with those administered chemotherapy ( P<0.001). The EFS of the allo-HSCT group was better than that of chemotherapy group, but the difference was not statistically significant ( P = 0.079). In the high miR-509 expression group, the allo-HSCT group had a better OS compared with that of the chemotherapy group ( P = 0.043). There was no significant difference in the EFS between the allo-HSCT group and chemotherapy group in high miR-509 group ( P = 0.154). Conclusions:The expression level of miR-509 may be helpful in the treatment selection of AML patients. Allo-HSCT can improve the prognosis of patients with low expression of miR-509.

Objective:To explore the effect of modified rope-ladder puncture and blunt-needle buttonhole puncture in new internal fistula of hemodialysis patients.Methods:From March 2019 and May 2020, a total of 70 patients undergoing maintenance hemodialysis in Heping Hospital Affiliated to Changzhi Medical College were enrolled using convenience sampling method and divided to the observation group (40 cases) and the control group (30 cases) by random number table method. The control group received blunt-needle buttonhole puncture and the observation group conducted modified rope-ladder puncture. The puncture situations, pain, hemostasis time after needle withdrawal, occurrence of puncture complications and internal fistula related complications were compared between the two groups.Results:There was no significant difference in success rate of one-time puncture between the two groups ( P>0.05) . The puncture pain of the observation group was severe than that of the control group ( P<0.05) , average pressing time at puncture sites was longer than that of the control group ( P<0.05) , and incidence of puncture complications was higher than that of the control group ( P<0.05) . Within one year of dialysis treatment, the incidence of internal fistula complications of the observation group was lower than that of the control group ( P<0.05) . Conclusions:The success rate of one-time puncture between modified rope-ladder puncture and blunt-needle buttonhole puncture are similar. The pain caused by blunt-needle buttonhole puncture is milder, hemostasis time is shorter and incidence of puncture complications is lower. However, the modified rope ladder puncture has less influence on the internal fistula function of patients.

Cashmere, also known as soft gold, is produced from the secondary hair follicles (SHFs) of cashmere goats. The number of SHFs determines the yield and quality of cashmere; therefore, it is of interest to investigate the transcriptional profiles present during cashmere goat hair follicle development. However, mechanisms underlying this development process remain largely unexplored, and studies regarding hair follicle development mostly use a murine research model. In this study, to provide a comprehensive understanding of cellular heterogeneity and cell fate decisions, single-cell RNA sequencing was performed on 19,705 single cells of the dorsal skin from cashmere goat fetuses at induction (embryonic day 60; E60), organogenesis (E90), and cytodifferentiation (E120) stages. For the first time, unsupervised clustering analysis identified 16 cell clusters, and their corresponding cell types were also characterized. Based on lineage inference, a detailed molecular landscape was revealed along the dermal and epidermal cell lineage developmental pathways. Notably, our current data also confirmed the heterogeneity of dermal papillae from different hair follicle types, which was further validated by immunofluorescence analysis. The current study identifies different biomarkers during cashmere goat hair follicle development and has implications for cashmere goat breeding in the future.

Recently, with the development and the continuous improvement of various CRISPR systems represented by CRISPR/Cas9, gene editing technology has been gradually improved, and widely applied to the preparation of animal models of human diseases. The gene edited animal models provide important materials for the study of pathogenesis, pathological process, prevention and treatment of human diseases. At present, the gene edited animal models used in human disease research include mainly the rodent models represented by mice and rats, and large animal models represented by pigs. Among them, rodents differ greatly from humans in all aspects of their bodies and have short life span as well, which cannot provide effective evaluation and long-term tracking for the research and treatment of human diseases. On the other hand, pig is closer to human in physiology, anatomy, nutrition and genetics, which provides an important animal model in the field of organ transplantation and human disease research. In this paper, the application of the gene edited animal models was summarized in the researches of 5 human diseases such as neurodegenerative diseases, familial hypertrophic cardiomyopathy, cancer, immunodeficiency diseases and metabolic diseases. We hope this paper will provide a reference for the research of human diseases and the preparation of relative animal models.
Animals ; CRISPR-Cas Systems ; Clustered Regularly Interspaced Short Palindromic Repeats ; Disease Models, Animal ; Gene Editing ; Humans

Next-generation sequencing has yielded a vast amount of cattle genomic data for global characterization of population genetic diversity and identification of genomic regions under natural and artificial selection. However, efficient storage, querying, and visualization of such large datasets remain challenging. Here, we developed a comprehensive database, the Bovine Genome Variation Database (BGVD). It provides six main functionalities:gene search, variation search, genomic sig-nature search, Genome Browser, alignment search tools, and the genome coordinate conversion tool. BGVD contains information on genomic variations comprising ～60.44 M SNPs, ～6.86 M indels, 76,634 CNV regions, and signatures of selective sweeps in 432 samples from modern cattle worldwide. Users can quickly retrieve distribution patterns of these variations for 54 cattle breeds through an interactive source of breed origin map, using a given gene symbol or genomic region for any of the three versions of the bovine reference genomes (ARS-UCD1.2, UMD3.1.1, and Btau 5.0.1). Signals of selection sweep are displayed as Manhattan plots and Genome Browser tracks. To further investigate and visualize the relationships between variants and signatures of selection, the Genome Browser integrates all variations, selection data, and resources, from NCBI, the UCSC Genome Browser, and Animal QTLdb. Collectively, all these features make the BGVD a useful archive for in-depth data mining and analyses of cattle biology and cattle breeding on a global scale. BGVD is publicly available at http://animal.nwsuaf.edu.cn/BosVar.

OBJECTIVETo observe the effects of a biotic elicitor fungal hyphae extract, an abiotic elicitor methyl jasmonate and their synergistic action on the accumulation of phenolic acids and tanshinones in Salvia miltiorrhiza hairy root.

METHODDifferent elicitors were added to S. miltiorrhiza hairy root, which was subcultured for 21 days, the dry weight and contents of phenolic acids and tanshinones were determined at different harvest-time.

RESULTS. miltiorrhiza hairy root growth was significantly inhibited by all three treatments and the accumulation of cryptotanshinone and dihydrotanshinone were promoted by each elicition. As for the accumulation of phenolic acids, there were differences between fungal elicitor and methyl jasmonate treatments, they were promoted by methyl jasmonate while inhibited in a certain extent by fungal hyphae extract.

CONCLUSIONFungal elicitor, methyl jasmonate and their synergistic action have significant influence on accumulation of components in S. miltiorrhiza hairy root, and the effect varies between phenolic acids and tanshinones. There is no correlation between production of water-soluble ingredients and fat-soluble components on the whole under three different treatments.

Acetates ; pharmacology ; Cyclopentanes ; pharmacology ; Diterpenes, Abietane ; metabolism ; Fungal Proteins ; metabolism ; pharmacology ; Hydroxybenzoates ; metabolism ; Membrane Glycoproteins ; metabolism ; pharmacology ; Oxylipins ; pharmacology ; Phytophthora ; chemistry ; metabolism ; Plant Extracts ; metabolism ; Plant Roots ; drug effects ; growth & development ; metabolism ; microbiology ; Salvia miltiorrhiza ; drug effects ; growth & development ; metabolism ; microbiology

OBJECTIVESalmonella isolates recovered from retail meats that were collected in supermarkets and free markets in Xi'an and Yangling areas of Shaanxi province were studied to determine antibiotic susceptibility.

METHODAntimicrobial susceptibility to 14 antibiotics of 193 salmonella isolates were determined by using agar dilution method, which was recommended by National Committee of Clinical Laboratory Standard (NCCLS), and E.coli ATCC25922 and E.faecalis ATCC29212 as standard control strains.

RESULTSThe 44.6% of the salmonella isolates were resistant to sulfamethoxazole, followed by resistance to kanamycin (40.9%), tetracycline (37.8%), amoxicillin (26.9%), ampicillin (25.4%), gentamicin (23.3%) and chloramphenicol (21.8%). Some isolates also showed resistance to fluoroquinolones, the rates for ciprofloxacin, enrofloxacin, levofloxacin and gatifloxacin were 22.3%, 21.8%, 20.8% and 21.2%, respectively. 55 isolates (28.5%) were multidrug resistant (MDR) strains, 28 of 193 isolates (14.5%) could resist at least 13 antibiotics, 24 isolates (12.4%) were resistant to from 4 to 12 antibiotics.

CONCLUSIONSalmonella isolates recovered from retail meats in Xi'an district of Shaanxi province were seriously resistant to antimicrobials commonly used as human and veterinary medicine.

Animals ; Cattle ; Chickens ; Drug Resistance, Multiple, Bacterial ; Food Microbiology ; Goats ; Meat Products ; microbiology ; Salmonella ; drug effects ; isolation & purification ; Sheep ; Swine