Objective:To purify HLA-DR molecules. Methods: Anti-HLA-DR antibody L243 was purified and coupled with CNBr activated Sepharose 4B gel. Immunoaffinity column was used to purify HLA-DR molecules. Results:Twenty micrograms of HLA-DR molecules were isolated from about 5 g Epstein-Barr virus-transformed human B lymphoblastoid cell line RAJI lysates by affinity chromatography. The purified HLA-DR molecules existed in α/β heterodimers form and could bind to conformation-dependent antibody L243. These HLA-DR molecules were separated into two strands,α and β,by boiling denaturation. These results are the basis for studying MHC Ⅱ binding peptide motif and CD4＋ T cell epitopes of antigens in future.

α-HgS is the main component of traditional Chinese medicine cinnabar, while β-HgS is the main component of Tibetan medicine Zuotai. However, there was no comparative study on the dissolution and absorption in gastrointestinal tract and bioaccumulation in organs of mercury in Cinnabar, Zuotai, α-HgS and β-HgS. In this study, the dissolution process of the four compounds in the human gastrointestinal tract was simulated to determine the mercury dissolutions and compare the mercury dissolution of different medicines and the dissolution-promoting capacity of different solutions. To explore the absorption and bioaccumulation of cinnabar and Zuotai in organisms, mice were orally administered with clinical equivalent doses cinnabar and Zuotai. Meanwhile, a group of mice was given α-HgS and β-HgS with the equivalent mercury with cinnabar, while another group was given β-HgS and HgCl2 with the equivalent mercury with Zuotai. The mercury absorption and bioaccumulation capacities of different medicines in mice and their mercury bioaccumulation in different tissues and organs were compared. The experimental results showed a high mercury dissolutions of Zuotai in artificial gastrointestinal fluid, which was followed by β-HgS, cinnabar and α-HgS. As for the mercury absorption and bioaccumulation in mice, HgCl2 was the highest, β-HgS was the next, and a-HgS was slightly higher than cinnabar. The organs with the mercury bioaccumulation from high to low were kidney, liver and brain. This study is close to clinical practices and can provide reference for the clinical safe medication as well as a study model for the safety evaluation on heavy metal-containing medicines by observing the mercury dissolution, absorption, distribution and accumulation of mercury-containing medicines cinnabar and zuotai.
Animals ; Brain ; metabolism ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Gastrointestinal Tract ; metabolism ; Kidney ; metabolism ; Liver ; metabolism ; Male ; Mercury ; chemistry ; pharmacokinetics ; Mercury Compounds ; chemistry ; pharmacokinetics ; Mice ; Solubility

To study the correlation between 50% tissue-culture infective dose (TCID50) value and p27 antigen S/P value of Avian leukosis virus subgroup J and discuss their significance, chicken embryo fibroblast (CEF) cells were inoculated with Avian leukosis virus subgroup J strain NX0101 and samples were tested continuously for ten days after changing maintenance media. The correlation between TCID50 and p27 antigen S/P value of ten days were then analysized. Simultaneously, DF-1 cells were inoculated with NX0101 and passaged to 20 generations. Samples taken from 1st generation, 5th generation, 10th generation, 15th generation and 20th generation were tested for the TCID50 titer and the p27 antigen S/P value separately. A significant Pearson correlation was found between them in CEF cells (r = 0.85277; P < 0.0001) and in DF-1 cells (r = 0.93000; P = 0.0220). This study provided an important parameter for predicting TCID50 by detecting the p27 antigen S/P value.
Animals ; Avian Leukosis ; virology ; Avian Leukosis Virus ; immunology ; pathogenicity ; Chick Embryo ; Fibroblasts ; virology ; Proliferating Cell Nuclear Antigen ; analysis ; immunology ; Viral Load ; immunology

Objective: To investigate the relationship between high sensitivity C-reactive protein (hs-CRP) level and incidence of left atrial spontaneous echocardiographic contrast (LASEC) in the patients with nonvalvular atrial fibrillation (AF). Methods: Four hundred and ninety consecutive patients with nonvalvular atrial fibrillation who underwent radiofrequency ablation for the first time from January 1, 2018 to June 30, 2018 in the Department of Cardiology, Beijing Anzhen Hospital were enrolled. According to the results of transesophageal echocardiography before radiofrequency ablation, patients were divided into the group without LASEC (n=338) and the group with LASEC (n=152). hs-CRP was determined by latex enhanced immunoturbidimetry. The relationship between hs-CRP and LASEC in patients with nonvalvular atrial fibrillation was investigated by univariate and multivariate logistic analysis. Results: LASEC was detected in 152 (31%) of 490 patients. Significant differences in age, type of atrial fibrillation, previous embolic events, fibrinogen, D-dimer, the left atrial anteroposterior diameter and CHA(2)DS(2)-VASc scores were found between patients with and without LASEC (all P<0.05). Compared with the group without LASEC, the serum hs-CRP level was significantly higher in the group with LASEC (3.16 (1.30, 5.23) mg/L vs. 0.67 (0.37, 1.48) mg/L, P<0.001). Multivariate logistic regression analysis showed that hs-CRP (OR=1.136, 95%CI 1.060 - 1.217, P<0.001) and D-dimer (OR=1.040, 95%CI 1.011 - 1.070, P=0.007) were independent determinants for LASEC in this patient cohort. Conclusions: hs-CRP is an independent determinant for LASEC in patients with nonvalvular atrial fibrillation. Inflammation may thus be involved in the formation of prethrombotic state in patients with nonvalvular atrial fibrillation.
Atrial Appendage ; Atrial Fibrillation/epidemiology* ; C-Reactive Protein ; Echocardiography, Transesophageal ; Electrocardiography ; Heart Atria ; Humans ; Incidence ; Risk Factors

Objective To investigate the current status of the coefficients variations (CVs) of internal quality control (IQC) data for serum procalcitonin in China.Methods Data had been collected by Web based submission system,the laboratories which enrolled in 2017 serum procalcitonin external quality assessment (EQA) program had attended.The data had includ ed:the CVs of two levels of IQC materials (level 1 and 2) in March of 2017 and long-term cumulative in control data.Mi crosoft Office Excel 2007 was used to analyze and process the data,the acceptable rates of CVs were calculated based on the 1/3TEa and 1/4TEa standards.The instruments which was used in laboratory internal quality control system of EQA,were grouped and counted,the acceptable rates of each group was calculated according to two evaluation standards.According to the laboratory detecting system was matched or not,to calculate the proportion of laboratories,and to adapt to the two standards.Results The acceptable rates of the same standards were close and the acceptable rates of level 2 were relatively higher.After grouping according to the instruments,the acceptable rates of each group were uneven.According to the labo ratory detecting system was matched or not,the acceptable rates of the matching system were much more higher.Conclusion To strengthen internal quality control system,and to improve the detection quality level much further.Laboratory should pay more attention to the mission of internal quality control,in order to ensure the reliability of test results.

Objective To investigate the internal quality control(IQC) of hemoglobin A2 (HbA2) and hemoglobin F (HbF) from 2014 to 2017 in China.Methods The results of IQC were collected from the laboratories which participated in external quality assessment (EQA) of National Center for Clinical Laboratories (NCCL) from 2014 to 2017,then the coefficient of variation (CV) was compared with 1/3TEa (6.67 ％),1/4TEa (5 ％).The proportion of laboratories meeting criteria were calculated to analyze IQC of HbA2 and HbF in China.The data were grouped based on the instruments used in laboratories,the acceptable rates of CVs of HbA2 and HbF in each group under two criteria in 2017 were calculated,respectively.Results In HbA2,more than 84％ of participant laboratories met 1/3TEa criteria and 70.83％ ～84.47％ of laboratories met 1/ 4TEa criteria.In HbF except for 2015,the more than 80％ laboratories whose month and cumulative CVs met 1/3TEa and 1/4TEa criteria accounted for 68.42 ％ ～ 85.07 ％,respectively.Under 1/3TEa and 1/4TEa criteria,sebia capillarys 2 instru ment and fully automatic hemoglobin analyzer bole Variant Ⅱ instrument group the acceptable rates of CVs above 85％,showed good precision for HbA2 and HbF detection.Conclusion At present,the precision level of HbA2 and HbF need to be further improved in laboratories of China,especially HbF.Laboratory should continue to strengthen the internal quality control,establish strict internal quality system to improve detection capacity.

Non-human primates (NHPs) are confirmed as reservoirs of Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi. In this study, 197 fresh fecal samples from 8 NHP species in Qinling Mountains, northwestern China, were collected and examined using multilocus sequence typing (MLST) method. The results showed that 35 (17.8%) samples were positive for tested parasites, including Cryptosporidium spp. (3.0%), G. intestinalis (2.0%), and E. bieneusi (12.7%). Cryptosporidium spp. were detected in 6 fecal samples of Macaca mulatta, and were identified as C. parvum (n=1) and C. andersoni (n=5). Subtyping analysis showed Cryptosporidium spp. belonged to the C. andersoni MLST subtype (A4, A4, A4, and A1) and C. parvum 60 kDa glycoprotein (gp60) subtype IId A15G2R1. G. intestinalis assemblage E was detected in 3 M. mulatta and 1 Saimiri sciureus. Intra-variations were observed at the triose phosphate isomerase (tpi), beta giardin (bg), and glutamate dehydrogenase (gdh) loci, with 3, 1, and 2 new subtypes found in respective locus. E. bieneusi was found in Cercopithecus neglectus (25.0%), Papio hamadrayas (16.7%), M. mulatta (16.3%), S. sciureus (10%), and Rhinopithecus roxellana (9.5%), with 5 ribosomal internal transcribed spacer (ITS) genotypes: 2 known genotypes (D and BEB6) and 3 novel genotypes (MH, XH, and BSH). These findings indicated the presence of zoonotic potential of Cryptosporidium spp. and E. bieneusi in NHPs in Qinling Mountains. This is the first report of C. andersoni in NHPs. The present study provided basic information for control of cryptosporidiosis, giardiasis, and microsporidiosis in human and animals in this area.
Animals ; China ; Cryptosporidiosis/*parasitology ; Cryptosporidium/classification/genetics/*isolation & purification ; Enterocytozoon/classification/genetics/*isolation & purification ; Feces/parasitology ; Female ; Genotype ; Giardia lamblia/classification/genetics/*isolation & purification ; Giardiasis/parasitology/*veterinary ; Male ; Microsporidiosis/parasitology/*veterinary ; Molecular Sequence Data ; Phylogeny ; Primate Diseases/*parasitology ; Primates/classification/parasitology

Objective To analyzed the differences between the status of internal quality control and industry standard of different platforms in second trimester prenatal screening,so as to provide recommendations to improve prenatal screening quality control.Methods Collected the coefficient of variation in control and accumulated coefficient of variation in control of 468 prenatal screening laboratory in September 2016,and the proportion of the laboratory that meet the industry standard and analyze the proportion of different platforms that meet the manufacturers standards and industry standards.Results For AFP,34.9 ％ ～ 100 ％ of the laboratory meeted the manufacturers standards and 25 ％ ～ 78 ％ meet the industry standards;for total HCG,59.2％ ～81.6％ of the laboratory meeted the manufacturers standards and 24.5 ％ ～30.6％ meeted the industry standards,for free β-hCG,38.9 ％ ～ 100 ％ of the laboratory meeted the manufacturers standards and 27.3 ％ ～ 63.0 ％ meeted the industry standards,for β-hCG,14.3％～68.8％ of the laboratory meeted the manufacturers standards and 14.3％～61.3 ％ meeted the industry standards.For uE3,19.7 ％ ～ 100％ of the laboratory meeted the manufacturers standards and 11.1~54.8％ meeted the industry standards.Conclusion Failure to meet the industry standards happens to all platforms.Prenatal screening laboratories need to pay attention to internal quality control to ensure the performance of the platforms meet the quality requirements of prenatal screening.

Objective: To investigate the molecular characteristics of ciprofloxacin-cefotaxime-azithromycin co-resistant Salmonella enterica serovar Thompson (S. Thompson) isolates from sporadic cases of foodborne diseases and aquatic foods in Hunan province. Methods: Ciprofloxacin-cefotaxime-azithromycin co-resistant S. Thompson isolates were selected from samples, and broth microdilution method was used to determine the resistance to 11 antibiotics of these isolates in vitro. Whole genome sequencing was used for investigating antimicrobial resistance gene patterns and phylogenetic relationships of strains. Results: Nine ciprofloxacin-cefotaxime-azithromycin co-resistant isolates were recovered from 19 S. Thompson isolates. Among nine ciprofloxacin-cefotaxime-azithromycin co-resistant isolates, eight of them harbored IncC plasmids, simultaneously carrying plasmid-mediated quinolone resistance (PMQR) genes qepA and qnrS1, β-lactamase resistance gene bla_CMY-2, azithromycin resistance gene mph(A), and one isolate harbored IncR plasmid, and carried PMQR genes qnrB4 and aac(6')-Ib-cr, bla_OXA-10 and mph(A). Genetic environment analysis showed that qnrS1, qepA, mph(A) and bla_CMY-2 genes might be integrated on genomes of strains by ISKra4, IS91, IS6100 and ISEcp1, respectively. Phylogenetic core genome comparisons demonstrated that ciprofloxacin-cefotaxime-azithromycin co-resistant isolates from patients and aquatic foods were genetically similar and clustered together. Conclusion: Ciprofloxacin-cefotaxime-azithromycin co-resistant S. Thompson isolates have been isolated from both human and aquatic food samples, suggesting that the spread of multidrug resistant Salmonella between human and aquatic animals.
Animals ; Humans ; Ciprofloxacin ; Cefotaxime ; Azithromycin ; Serogroup ; Phylogeny ; Drug Resistance, Multiple, Bacterial/genetics* ; Anti-Bacterial Agents/pharmacology* ; Salmonella ; Quinolones ; Foodborne Diseases ; Plasmids ; Salmonella enterica ; Microbial Sensitivity Tests

As a ligand-dependent transcription factor,retinoid-associated orphan receptor γt(RORyt)that controls T helper(Th)17 cell differentiation and interleukin(IL)-17 expression plays a critical role in the pro-gression of several inflammatory and autoimmune conditions.An emerging novel approach to the therapy of these diseases thus involves controlling the transcriptional capacity of RORyt to decrease Th17 cell development and IL-17 production.Several RORyt inhibitors including both antagonists and inverse agonists have been discovered to regulate the transcriptional activity of RORyt by binding to orthosteric-or allosteric-binding sites in the ligand-binding domain.Some of small-molecule inhibitors have entered clinical evaluations.Therefore,in current review,the role of RORyt in Th17 regulation and Th17-related inflammatory and autoimmune diseases was highlighted.Notably,the recently developed RORyt inhibitors were summarized,with an emphasis on their optimization from lead compounds,ef-ficacy,toxicity,mechanisms of action,and clinical trials.The limitations of current development in this area were also discussed to facilitate future research.