Objective To evaluate the repair of benign constriction of the porta hepatic duct by pedicled autograft. Methods The clinical data of repair by pedicled valve of gallbladder in 38 cases, by pedicled gastric wall in 2 cases and by round ligament of the liver in 2 cases were retrospectively analyzed. Results There were no operative mortality and no complications such as bile leaks and hemorrhage. Cholangiography through T-tube showed patency in all postoperative patients. A follow-up from 6 months to 14 years in 40 out of 42 cases found cholangitis in 2 patients with good response to conservative therapy. Conclusions Treatment of benign stricture of porta hepatic duct by pedicled autograft is physiologic, convenient and effective.

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

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

OBJECTIVETo study the chemical constituents of Polyporus ellissi.

METHODSilica gel column chromatography was applied for the isolation and purification of the constituents. The structures were established by means of spectroscopic and chemical data.

RESULTSix compounds were obtained and identified as cerebroside B (I), cerebroside D (II), ergosterol peroxide (III), 9(11)-dehydroergosterol peroxide (IV), mannitol (V) and palmitate-1-glycerol (VI).

CONCLUSIONCompounds (I) and (II) were isolated from the genus Polyporus for the first time.

Cerebrosides ; chemistry ; isolation & purification ; Mannitol ; chemistry ; isolation & purification ; Molecular Structure ; Polyporaceae ; chemistry

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*

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

Male germ stem cells (mGSCs), which is in testis after sex differentiation, derive from primordial germ cells. In this study, bovine mGSCs were isolated from testis of 20 weeks fetuses. Number of CD9 positive cells of the cells through two-steps adhering plates velocity different was 95.8% by flow cytometer. The carina-type cells clones and the plane-type cells clones appeared in co-cultured system. One cells lines had been successively maintained for 4 passages, and the cells clusters showed AKP positive staining. The cells clusters showed nest-shape in third passage showed SSEA1 and Oct-4 positive staining. These cells can also spontaneously differentiate into c-kit positive staining germ cells, and the cells were directional induced to formaactin positive staining cardiac-like cells cluster and NF positive staining neuron-like cells. The conclusion showed that male germ stem cells from 20 weeks bovine fetuses could be in vitro formed like embryonic stem cells.
Animals ; Cattle ; Cell Differentiation ; physiology ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; Fetus ; cytology ; Male ; Pluripotent Stem Cells ; cytology ; Spermatozoa ; cytology

It is very easy for the pro-UK to lose it's biological activity because of the digestion of pro-UK by the thrombin or the inhibition of pro-UK by the PAI-I. So three pro-UK mutant (pro-UK) genes were constructed in this experiment with the PCR point-mutant method. The thrombin cleavage site Arg156 in pro-UK was mutated into His156, and named as pro-UKM1; PAI binding sites Arg178, Arg179, Arg181 were mutated into Lys178, Lys179, His181, named as pro-UKM2; The mutant containing His156, Lys178, Lys179, His181 as pro-UKM3. Three mutants were expressed in CHO cells respectively and analyzed with SDS-PAGE fibrin plate assay and western blot. The results showed that the three mutants and the native pro-UK have the same single electrophoresis band indicating most of the pro-UK was single chain. In vitro plasma clot lysis assays indicated that the pro-UKM1 have the ability to resistant against thrombin digestion; pro-UK2 could resist against PAI inhibition; while pro-UK3 improved resistances against both thrombin and PAI. It looks very promising that the pro-UK3 can be a new medicine of dissolving thrombus.
Animals ; Base Sequence ; Blotting, Western ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Electrophoresis, Polyacrylamide Gel ; Humans ; Molecular Sequence Data ; Mutant Proteins ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Urokinase-Type Plasminogen Activator ; biosynthesis ; genetics

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.

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.