Objective To study the relevance of 1iver fibrosis at Kasai procedure with age, degree of pathological changes of liver tissue, header bile ductuler proliferation and pathological feature of portal fibrous tissue in biliary atresia patients. Methods From January 2013 to June 2016, 48 biliary atresia patients accepted Kasai procedure, and liver tissue biopsy and portal fibrous tissue excision was done. Level of liver fibrosis was evaluated under microscope using Masson trichrome staining and HE staining, and extent of ductular reaction was evaluated using CKl9 immunohistochemistry staining with image analysis technology. Based on classification of Okamoto severity degree of liver fibrosis, forty eight patients were divided into two groups:low-grade liver fibrosis group (23 patients)and severe liver fibrosis group (25 patients). The relevance of age at Kasai procedure, liver pathological changes, header bile ductuler proliferation, portal fibrous tissue in volume and bile ductuler proliferation, inflammation reaction with severity degree of liver fibrosis was analyzed with SPSSl6.0 software statistically. Results Age of severe 1iver fibrosis group at Kasai procedure were 38- 89 d, and the mean age was (62.64 ± 14.92) d. Age of low-grade 1iver fibrosis group were 18- 92 d, and the mean age was (53.13 ± 16.89) d. There was significant difference (t=2.071, P=0.044). Mean number of patients with header bile ductuler proliferation at Kasai procedure in severe 1iver fibrosis group and low-grade 1iver fibrosis group were (17.28 ± 4.31) cases and (11.39 ± 4.00) cases, and there was significant difference (t=4.900, P=0.000). There was significant difference in degree of hepatocellular pathological changes at Kasai procedure between groups (χ2 =4.691, P=0.030). There were no significant differences in the volume of portal fibrous tissue in volume (t=0.276, P=0.784), portal bile ducts proliferation (t=0.53, P=0.60) and inflammation reaction (χ2=3.074, P=0.080) between groups. Conclusions There is significant relevance between the degree of 1iver fibrosis in biliary atresia patients at Kasai procedure and age, level of hepatocellular pathological change and degree of header bile ductuler proliferation , The more older the age, the more sever the hepatocellular pathological change and the bile ductuler proliferation, the more sever the liver fibrous.

Prone position ventilation has become an important part of lung protective ventilation in acute respiratory distress syndrome (ARDS) patients. Timely and appropriate implementation can improve the mortality of such patients. The same is true for patients with critical coronavirus disease 2019 (COVID-19) patients, but safety assessment, control of implementation details and prevention of complications still need to be further standardized during the practical procedure of prone position ventilation. The purpose of this paper is to explain the specific requirements and key points of prone position ventilation in critical COVID-19 patients, in order to promote the application of prone position ventilation in the treatment of patients with critical COVID-19 and reduce the occurrence of related complications.

This study focused on the intestinal absorption of traditional Chinese medicines (TCM) to reveal the scientific connotation of the compatibility of TCM pairs. The single pass intestinal perfusion (SPIP) was used in rats to compare the absorption of single extracts from Puerariae Lobatae Radix, single extracts from Ginseng Radix et Rhizoma, combined extracts from Puerariae Lobatae Radix and Ginseng Radix et Rhizoma and Puerariae Lobatae Radix and Ginseng Radix et Rhizoma mixture in rats. The content of puerarin, ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 in liquid were tested by HPLC. The speed constant (Ka) and apparent permeability coefficients (Papp) were calculated and compared. Specifically, the order of puerarin Ka and Papp values from high to low was Ginseng Radix et Rhizoma and Puerariae Lobatae Radix mixture > single extracts from Puerariae Lobatae Radix > combined extracts from Ginseng Radix et Rhizoma and Puerariae Lobatae Radix; the order of ginsenosides Ka and Papp values from high to low was Ginseng Radix et Rhizoma and Puerariae Lobatae Radix mixture > single extracts from Ginseng Radix et Rhizoma > combined extracts from Ginseng Radix et Rhizoma and Puerariae Lobatae Radix. The combined administration of Ginseng Radix et Rhizoma and Puerariae Lobatae Radix may improve the absorption in the intestinal tract.
Animals ; Ginsenosides ; pharmacokinetics ; Intestinal Absorption ; Isoflavones ; pharmacokinetics ; Male ; Medicine, Chinese Traditional ; Panax ; chemistry ; Plant Extracts ; pharmacokinetics ; Pueraria ; chemistry ; Rats ; Rats, Sprague-Dawley ; Rhizome

Objective To evaluate the clinical performance of AQT90 FLEX,a novel time-resolved fluorescence based point-of-care test (POCT) for quantification of D-dimer in elderly patients.Methods The method from Quantitative D-dimer assay (WS/T 477-2015) for testing equipment performance was used as a reference to evaluate the clinical performance of AQT90 FLEX.The correlation was compared between testing results of D-dimer using the AQT90 immune-assay analyzer and those using the ACL TOP coagulation analyzer.Results At high concentration of D-dimer,the within-run precision coefficient of variation (CV)was 2.619％,and at low concentration of D-dimer,the within-run precision CV was 2.767％.The pollution-carrying rate was 0.12％.The measured data from AQT90 and ACL TOP had a correlation coefficient of r =0.9491 (P ＜ 0.01).The equation of the line of best fit for D-dimer with which all AQT90 results can be adapted to the ACL TOP was:AQT90 =2.52 ACL TOP + 0.15.The number from the equation was slightly greater in female than that in male,and it was also increased in elderly.Conclusions The AQT90 FLEX had rational precision and linearity in determination of concentration.There was a high agreement between the testing results from AQT90 and those from ACL TOP.It was recommended to use a slope of 2.52 and an intercept of 0.15 to adjust the D-dimer values of the ACL TOP to the AQT90 FLEX assay systems.POCT for D-dimer by AQT90 FLEX raises feasibility for use in elderly patients.

Objective • To investigate the reliability and validity of Chinese version of Working Alliance Inventory-Short Form Revised (WAI-SR) in the hospitalized schizophrenics. Methods • A sample of 124 inpatient schizophrenics who were diagnosed according to International Classification of Diseases 10th Revision in Shanghai Mental Health Center and collected by convenience sampling, were interviewed by questionnaire. Item analysis, internal consistency, construct validity and criterion validity were tested using total sample. Patient-Doctor Relationship Questionnaire-15 (PDRQ-15) and Chinese version of Wake Forest Physician Trust Scale (WFPTS) were used as criteria to test criterion validity. Results • Exploratory factor analysis extracted two dimensions labeled "goal and task" and "affective bond", and the cumulative variance contribution rate was 63.99%. Confirmatory factor analysis showed that the two-factor model had higher adjusted goodness of fit index. The scores of scale and its two dimensions exhibited positive correlations with PDRQ-15 and WFPTS (r=0.324-0.726, P<0.01). The internal consistency of the scale (Cronbach's α coefficient) was 0.864, and split-half reliability was 0.919.Conclusion • The Chinese version of WAI-SR is acceptable in reliability and validity and can be recommended for alliance assessment in the inpatients with schizophrenia.

Objective To evaluate the immunogenic stability and hereditary stability of Neisseria meningitides serogroup W135/Y[CMCC(B)29037/CMCC(B)29028]within all the passages,which isolated from china.Methods The toxicity of the 3rd,5th,10th,15th,20th,25th and 30th passage of the Neisseria meningitidis was assayed in mice.Serological detection and biochemical detection were measured,and immunized mice subcutaneously.The antigeeicity of each passage of Neisseria meningitides serogroup W135/Y were measured by serum bactericidal test and the indirect ELISA.With the 30 passage of Neisseria meningitides serogroup W135/Y,the effect to the encephalic tissue was measured in mice.Fermented the Neisseria meningitides serogroup W135/Y with 30 passage and purified the capsular polysaccharide,then analyzed the quality respectively.Results The LD50 of the strains CMCC(B)29037/29028 of each passage was low(LD50 ≥ 109),and all the 30logical detection and all the 30 passage of the two strains were half in the tube agglutination.Glucose and maltose fermentation test were positive.Fructose,sucrose and lactose fermentation test were negative.The GMT of immunogenicity were 1114 and 2229 respectively and all the 30 passage were more than 640 and 1040 respectively.After Immunization with individual 30 passage of the Neisseria meningitides,the titer in serum bactericidal assay(SBA)and indirect ELISA were no difference.The capsular polysaccharide purified from Neisseria meningitides serogroup W135/Y met the quality standard.Conclusion Neisseria meningitides serogroup W135/Y,CMCC(B)29037/29028,used in the manufacture of the meningococcal conjugate vaccine,are stable in the toxicity,antigenicity,immunogenicity.Serological detection and biochemical detection are qulified,and the capsular polysaccharide has met the quality standard.

HPV16 L2E7 is a fusion protein used for therapeutical vaccine targeting HPV virus. To increase its expression in Escherichia coli, we optimized the codon usage of HPV16 l2e7 gene based on its codon usage bias. The optimized gene of HPV16 sl2e7 was cloned into three different vectors: pGEX-5X-1, pQE30, ET41a, and expressed in JM109, JM109 (DE3) and BL21 (DE3) lines separately. A high expression line was selected with pET41a vector in BL21 (DE3) cells. After optimization of the growth condition, including inoculation amount, IPTG concentration, induction time and temperature, the expression level of HPV16 L2E7 was increased from less than 10% to about 28% of total protein. HPV16 L2E7 protein was then purified from 15 L culture by means of SP Sepharose Fast Flow, Q Sepharose Fast Flow and Superdex 200 pg. After renaturing, HPV16 L2E7 protein with ≥ 95% purity was achieved, which was confirmed via SDS-PAGE gel and Western blotting. The combined use of purified HPV16 L2E7 and CpG helper has shown clear inhibition of tumor growth in mice injected with tumor cells, with six out of eight mice shown no sign of tumor. This study lays a solid foundation for a new pipeline of large-scale vaccine production.
Animals ; Capsid Proteins ; biosynthesis ; Codon ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Genetic Vectors ; Human papillomavirus 16 ; Mice ; Neoplasms, Experimental ; prevention & control ; Oncogene Proteins, Viral ; biosynthesis ; Papillomavirus E7 Proteins ; biosynthesis ; Papillomavirus Vaccines ; therapeutic use ; Recombinant Fusion Proteins ; biosynthesis

We established an ELISPOT for bovine interferon-gamma (BoIFN-γ), and applied it in the diagnosis of bovine tuberculosis (bTB). Monoclonal antibodies that can bind with native BoIFN-γ were screened as the coating antibody and detecting antibody. After optimization of detecting conditions including coating antibody concentration, cell number, and detecting antibody concentration, the ELISPOT assay was established. Peripheral mononuclear cells (PBMCs) isolated from 30 cows were co-cultured with PPD, and detected with the ELISPOT assay. The optimal conditions of ELISPOT assay were 2.5 μg/mL coating antibody 2G5, 2.5 x 10(5) cells/well, and 1 μg/mL detecting antibody Bio-5E11. In these 30 cows tested both with the ELISPOT assay and the BOVIGAM kit, 11 cows were proved to be positive in ELISOPT assay with the sensitivity of 78.6%, and 12 cows were proved to be negative in ELISOPT assay with the specificity of 75%. The ELISPOT assay for BoIFN-γ could be used to detect bTB efficiently and it might be an alternative method for the diagnosis of bTB.
Animals ; Antibodies, Monoclonal ; Cattle ; Enzyme-Linked Immunospot Assay ; veterinary ; Female ; Interferon-gamma ; isolation & purification ; Sensitivity and Specificity ; Tuberculosis, Bovine ; diagnosis

AIM: To discuss the protective effects and possible mechanisms of 17β-estradiol on human retinal pigment epithelial ( RPE) cells induced by high glucose. ·METHODS: RPE cells were cultured and divided into four groups according to randomized controlled method:blank control group:the cells were treated with 5. 5mmol/L routine glucose medium for processing; high glucose group: cells were treated with 100mmol/L glucose for 12h;17β-estradiol low concentration group: after treated with 10 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h; 17β-estradiol high concentration group: after treated with 100 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h. Cell viability were tested by MTT colorimetric detection. Cells apoptosis were detected by Hochest33258 staining. Intracellular reactive oxygen species( ROS) level were detected by H2 DCFDA staining. Expression of CAT, SOD and MDA were tested by colorimetric detection. · RESULTS: RPE cell activity decreased with the concentration of glucose increased; 17β-estradiol inhibited high glucose-induced cell viability decrease in RPE cells, decreased the apoptosis rate of RPE cells and intracellular ROS generation; besides, 17β-estradiol significantly increased the expression of CAT, SOD and decreased the expression of MDA in RPE cells. ·CONCLUSION: The 17β-estradiol effectively inhibited high glucose -induced RPE cells damage, which provide reliable experimental basis for the treatment of injuries in RPE cells.

BACKGROUND: Henoch-Schonlein purpura (HSP) is an immune complex-mediated disease predominantly characterized by the deposition of circulating immune complexes containing immunoglobulin A (IgA) on the walls of small vessels. Although the pathogenesis of HSP is not yet fully understood, some researchers proposed that B-cell activation might play a critical role in the development of this disease. OBJECTIVE: To investigate the serum levels of visfatin (pre-B-cell colony-enhancing factor), B-cell-activating factor (BAFF), and CXCL13, and to analyze their association with disease severity. METHODS: The serum levels of visfatin, BAFF, and CXCL13 were measured by using a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) in 43 patients with HSP and 45 controls. The serum levels of IgA anticardiolipin antibodies (ACA) were detected by using a double-antigen sandwich ELISA. RESULTS: Levels of visfatin but not BAFF and CXCL13 were significantly elevated in the sera of patients with HSP in the acute stage, and restored to normal levels in the convalescent stage. Furthermore, serum levels of visfatin were significantly higher in patients with HSP having renal involvement than in those without renal involvement. Serum levels of visfatin were correlated with the severity of HSP and serum concentration of ACA-IgA. CONCLUSION: We show for the first time that the serum levels of visfatin are abnormally elevated in patients with HSP. Visfatin may be associated with the pathogenesis of HSP.
Antibodies, Anticardiolipin ; Antigen-Antibody Complex ; B-Cell Activating Factor ; B-Lymphocytes ; Chemokine CXCL13 ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin A ; Nicotinamide Phosphoribosyltransferase* ; Purpura, Schoenlein-Henoch*