Objective To investigate the serum levels of soluble human leukocyte antigen (sHLA)-G in patients with polymyositis (PM) or dermatomyositis (DM),and to analyze its association with clinical features and possible role in the pathogenesis of PM/DM.Methods Serum sHLA-G levels of 26 patients with PM,70 patients with DM and 35 matched healthy controls were measured by ELISA.The relationship between the sHLA-G levels and clinical features or seroimmunological data in the patients with PM/DM was analyzed.Results Serum levels of sHLA-G in PM/DM patients were significantly higher compared to healthy controls [(44±70) U/ml vs (4±5) U/ml,P＜0.01].There was statistically significant difference between DM patients and PM patients [(54±81) U/ml vs (27±41) U/ml,P=0.004].The incidence of dysphagia was significantly higher in sHLA-G elevated group than those in sHLA-G normal group (P=0.001).Additionally,Spearman rank correlation analysis showed that the serum sHLA-G levels were positively correlated with serum C3 (r=0.284,P=0.021),but negatively correlated with CD3+ T cells (r=-0.233,P=0.047) and CD4+ T cells (r=-0.287,P=0.015) in the peripheral blood in patients with PM/DM.Serum levels of sHLA-G in non-treated PM/DM patients were significantly higher compared to treated patients [(77±99) U/ml vs (34±52) U/ml,P=0.021].No relationship between serum sHLA-G levels and PM/DM disease activity,or different drug therapy was found.Conclusion Serum levels of sHLA-G are increased in PM/DM patients.The increased production of sHLA-G,paralleled with higher incidence of dysphagia and lower level of CD3+ T cells and CD4+ T cells,indicates that sHLA-G may play an important role in the pathogenesis of PM/DM.

Objective This study aims to test the hypothesis that low density granulocytes (LDGs) is involved in the pathogenesis of DM associated-Interstitial lung disease (ILD).Methods Forty eight DM patients (28 with ILD) and 19 age-and sex-matched healthy Chinese volunteers were recruited to this study.LDGs percentage in peripheral blood mononuclear cells (PBMCs) was tested by flow cytometry.Neutrophilrelated genes (LL-37,MPO and MMP-8) expressions in PBMCs were tested by quantitative RT-PCR.Myositis disease activity assessment visual analogue scales (MYOACT) was used to assess the disease activity.Percentages of LDGs were compared in patients with ILD and without by using unpaired t test with Welch's correction,the correlations between LDGs and clinical parameters were further analyzed by linear correlation analysis.The expressions of neutrophil-related mRNA and proteins in PBMCs were compared by using MannWhitney U test.Results LDGs percentage in PBMCs was 7.1-fold higher in DM patients than healthy controls [(9.1±11.5)％ vs (1.3±0.7)％,t=4.664,P＜0.01].LDGs percentage in PBMCs was 2.7-fold higher in DM patients with ILD than DM patients without ILD [(12.3±14.1)％ vs (4.5±2.6)％,t=2.835,P=0.008 3].The mRNA expression level of LL-37,MPO and MMP-8 and LL-37 protein levels in the DM group were significantly higher than those in the control group.LDGs percentage positively correlated with MYOACT lung disease activity scores (r=0.439,P=0.010).Conclusion Percentage of LDGs in PBMCs is significantly increased in DM patients with ILD and positively correlated with MYOACT lung disease activity scores,suggesting that abnormall increasing of LDGs is a potential contributor to the pathogenesis of DM-associated ILD.

matK is one of the core DNA barcode markers for plant DNA barcode identification and its universality using single makers has been in controversy. However, the universalities of different matK primer pairs in same seed plant group (order) and same matK primer pairs in different seed plant groups (order) are lack of systematic research. In this study, we collected 14563 full-length matK sequences of 11429 species of 3292 genera in 239 families belonging to 36 orders in seed plants. The universalities of 13 matK primer pairs and its 78 primer com-binations have been assessed using bioinformatics methods. The results indicated that xf/5r, 1F/8R, 390F/1326R and 3F_KIM/1R_KIM were the four most universality primer pairs. The four markers' universalities were 91.18%, 84.65%, 79.81% and 80.94% respectively in all 11429 seed plants. The most universality primer pairs in different orders were different. For each order, the primer pair with maximum universality was different. the xf/5r was the basal primer pair for primer combination and 1F/8R, 1F/1R, M3/M4 and 3F_KIM/1R_KIM could be the complementary primer pairs. This study could be a valuable resource for the primer selection of the research DNA barcoding identification in seed plants.

Abstract] Objective To investigate the in vivo therapeutic effects of an extract of herb medi-cines, YiGanQingDuKeLi, in combination with adefovir dipivoxil (ADV) on the rebound of duck hepatitis B virus ( DHBV) multiplication after withdrawal of ADV treatment.Methods Peking ducks were infected with DHBV positive serum samples for 7 days and then screened by SYBR Green real-time PCR.The ducks positive for DHBV were randomly divided into five groups including the model control group, the ADV treat-ment group, the herb treatment group, the high-dose combination therapy group and the low-dose combina-tion therapy group.The ducks in the ADV treatment and the herb treatment groups were respectively treated with distilled water and YiGanQingDuKeLi (1.2 g/ml) for 14 days after the treatment of ADV (0.25 mg/ml) for 21 days.The ducks in the high-dose group were treated with YiGanQingDuKeLi (1.2 g/ml) for 14 days after the combined treatment with high-dose YiGanQingDuKeLi (1.2 g/ml) and ADV (0.25 mg/ml) for 21 days.The ducks in the low-dose group were treated with YiGanQingDuKeLi (0.6 g/ml) for 14 days after the combined treatment with YiGanQingDuKeLi (0.6 g/ml) and ADV (0.125 mg/ml) for 21 days.Blood samples were collected from each duck via leg vein after 0, 7, 14 and 21 days of drug adminis-tration and after 7 and 14 days of drug withdrawal.The levels of DHBV-DNA, alanine aminotransferase ( ALT) and aspartate aminotransferase ( AST) in blood serum samples were detected.Results Compared with the model group, the levels of DHBV-DNA, ALT and AST in ducks from the herb treatment group and combined treatment groups were decreased before the discontinuation of ADV treatment ( P<0.05 or P<0.01).Moreover, the titers of DHBV-DNA in ducks treated with high doses of drugs were much lower than those from ADV treatment group.The levels of DHBV-DNA, ALT and AST in ducks treated with herb medi-cine and high doses of drugs remained at relatively low levels after the cessation of ADV treatment, but re-bounded significantly in ducks with ADV treatment.The levels of DHBV-DNA and ALT rebounded slightly in ducks treated with low doses of drugs as compared with those of ADV treatment group ( P<0.01 or P<0.05).Conclusion The treatment of YiGanQingDuKeLi in combination with ADV could inhibit not only the in vivo replication of DHBV, but also the rebound of DHBV multiplication after ADV withdrawal.

Objective This study was focused on the association between neutrophil extracellular traps (NETs) and interstitial lung disease (ILD) in patients with dermatomyositis (DM).Methods Thirty six patients who satisfied the Bohan & Peter criteria for DM were recruited to this study,among whom 19 were complicated with ILD.Forty seven age and sex matched healthy Chinese volunteers were selected to be control subjects.The plasma samples of these patients were tested for the formation and degradation of NETs.Results DM plasma induced more NETs formation than control plasma did [(246±93) RFUs vs (192±53) RFUs,P=0.002].Compared to control,DM plasma exhibited a signficantry decreased ability to degrade NETs.Further mere,compared with DM patients without ILD (DMNL),DM patients with ILD (DML) could not degrade NETs completely [(83±13)％ vs (59±21)％,P＜0.01].All four DM patients with subacute ILD exhibited a significantly lower ability to degrade NETs than patients with chronic or asymptomatic ILD [(36±14)％ vs (65±19)％,P=0.0139].Conclusion These data show that more NETs formation is induced by plasma and DML fails to completely degrade NETs.These suggest that NETs may play a role in the pathogenesis of DM and DM-associated ILD.

Aim To screen out the antigenic sequences from HCV core protein random peptide libraries displayed on phage and to explore a new way to screen the viral antigens. Methods The anti-HCV core antibody-positive serum was used to screen antigenic peptides from the HCV core protein random peptide libraries displayed on phage for 4 rounds. Detection of numbers of positive clones, positive rate of insertion of HCV random DNA and positive rate of hybridization with HCV core probes were used to evaluate the screening effects. The DNA sequences of 7 selected clones with positive hybridization were determined and analysed. Results Six out of 7 sequences are HCV core protein sequences, in which 5 were perfectly displayed,and one was possibly displayed. These sequences included several major HCV core antigenic epitopes. The remaining one was E.coli nrfa gene. Conclusion The phage display technique can be applied to study the viral antigenic peptides with the advantages of simple, accuracy and rapidity.

OBJECTIVETo study the effect of progesterone on interleukin-6 (IL-6) release from human umbilic vein endothelial cells (HUVECs) induced by high mobility group box-1 protein (HMGB1).

METHODThe recombinant expression plasmid pET14b-HMGB1 was constructed and transformed into competent E.coli BL21 cells to obtain HMGB1 protein, which was purified with chromatography on Ni-NTA Sepharose column. Cultured HUVECs were treated with purified HMGB1 protein alone at the concentrations 0, 10, 100, 500, and 1000 ng/ml, progesterone alone at the concentrations of 0, 0.1, 1, 10, 100 mmol/L, or with both HMGB1 protein (500 ng/ml) and progesterone at the terminal concentrations of 0, 0.1, 1, 10, and 100 mmol/L. Twenty-four hours later, the supernatant of the cell culture medium was collected to detect the levels of IL-6 using enzyme-linked immunosorbent assay (ELISA).

RESULTSThe IL-6 levels in HUVEC culture medium was slightly decreased after treatment with low-concentration HMGB1 but increased obviously following treatment with high-concentration HMGB1, and these effects could be dose-dependently inhibited by progesterone. Progesterone alone did not result in any noticeable changes of IL-6 levels in the cell culture medium.

CONCLUSIONSProgesterone can dose-dependently inhibit HMGB1-induced IL-6 release from HUVECs, suggesting the protective role of progesterone in endotoxemia.

Cells, Cultured ; Dose-Response Relationship, Drug ; Endothelial Cells ; cytology ; metabolism ; HMG-Box Domains ; HMGB1 Protein ; pharmacology ; Humans ; Interleukin-6 ; biosynthesis ; Progesterone ; pharmacology ; Umbilical Veins ; cytology ; metabolism

OBJECTIVETo investigate the cloning and expression of Treponema pallidum (TP) specific antigen TpN17 and an epitope of TpN44.5 and the clinical application of this fusion antigen.

METHODSTpN17 gene was amplified from the genome of TP and an epitope of TpN44.5 was spliced to the 5' end of TpN17 gene. This modified TpN17 gene was cloned into the expression vector pGEX4T-2. The recombinant fusion antigen was purified by affinity chromatography and then an enzyme-linked immunosorbent assay (ELISA) kit was prepared. With this kit, the sera of 10 normal persons and 10 TP patients were tested.

RESULTSThe molecular weight of the purified fusion antigen was 45,000. Tested with ELISA, 10 serum samples of the TP patients were positive and another 10 of the normal persons were negative. ELISA equipped with the GST-epi-TpN17 antigen showed higher sensitivity and specificity as compared with routine methods.

CONCLUSIONThe recombinant fusion TP specific antigen GST-epi-TpN17 was suitable for the preparation of ELISA kit in clinical examinations.

Antigens, Bacterial ; biosynthesis ; genetics ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunodominant Epitopes ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; biosynthesis ; Sensitivity and Specificity ; Syphilis ; diagnosis ; Treponema pallidum ; immunology

Objective To understand the evolutionary characterization of hemagglutinin (HA)gene of pandemic H1N1 influenza virus in Guangdong during 2009-2011. MethodsWe selected 83 pandemic H1N1 strains isolated in Guangdong during 2009-2011. The HA1 genes were sequenced and analyzed comparatively by Bioedit 7.0 and MEGA 4.0. ResultsThe evolutionary rate of Hal gene of pandemic H1N1 and seasonal H1N1 viruses was 5.2×10-3 substitutions/site/year, higher than that of seasonal H1N1 viruses. Most amino acid changes in HA1 molecules accumulated on the surface of the molecule and were partly located in antigenic sites. Two fatal infections were detected with a mutation at HA residue 222, in one virus with a change D222G, and in one virus D222N. ConclusionThe phylogenetic analysis demonstrates that the influenza epidemic in Guangdong at the beginning of 2011 are due to occurrence of genetic changes of pandemic H1N1 virus. The amino acid change at residue 222 of the HA1 are likely to be associated with severe or even fatal illness.

Objective: To explore the application present situation of the WeChat public accounts of colleges psychological health education in Hunan Province, and to provide a scientific reference for strengthening the construction of WeChat public accounts of colleges psychological health education. Methods: The WCI(WeChat Communication Index) of the WeChat public accounts of college psychological health education in Hunan province, the numbers of reading and watching of the push articles which were published during period between June 1 and June 30 in 2019 were collected through the Qingbo data platform. The characteristics of the top 10 articles in reading numbers were analyzed. Results: Totally 19 universities in Hunan Province opened WeChat public accounts of psychological health education, accounting for 51.16% of all universities in Hunan Province. In June 2019, the average WCI of the WeChat public account of college psychological health education in Hunan Province was(159.3±81.3), with 267 articles and average(15.7±14.9). The total number of reading was 38 921, and the average reading number of the articles was 145.8. The total number of watching was 607, and the average watching number of articles was 2.3. Eight articles of the the top 10 articles in reading number were from the top 3 WeChat public accounts in WCI. Four articles were about online and offline interaction and the content of the other 6 articles combined with hot current events, closed to college students’ lives in the top 10 articles in reading numbers. Two articles of the the top 10 articles in reading numbers were film. Conclusion The proportion of the universities which have opened the WeChat Public account of psychological health education in Hunan Province is not very high, and the dissemination of the WeChat Public accounts of college psychological health education is not very strong. The construction quality of WeChat public account is an important factor affecting the reading number of push article. The articles with content combining with current hot events and closing to college students’ life have higher reading number than the other articles. Video work are more popular than text articles.