Objective To investigate the characteristics of CD8+ stem memory T cells (CD8+Tscm) in patients with chronic HIV-1 infection before and after antiretroviral therapy (ART) and to analyze their associations with progression of HIV-1 infection.Methods Thirty-six patients with chronic HIV-1 infection and 20 healthy subjects were enrolled in this study.Flow cytometry was performed to detect the percentages and absolute numbers of CD8+Tscm in patients with chronic HIV-1 infection before and after antiretroviral therapy (ART) as well as in healthy subjects.Correlation analysis was used to demonstrate the relationships between CD8+Tscm and markers for progression of HIV-1 infection (CD4+T cell count, HIV-1 viral load and level of activated T cells).Results The percentages and the absolute numbers of CD8+Tscm in patients with chronic HIV-1 infection had no significant change before and after ART.They were respectively positively correlated with the percentages and the absolute numbers of CD4+Tscm.The percentage of CD8+Tscm was proportional to the percentage of CD8+ central memory T cells (CD8+Tcm), but was inversely proportional to the percentage of CD8+ effector memory T cells (CD8+Tem).In addition, the percentages of CD8+Tscm in patients with HIV-1 infection were negatively correlated with the viral loads before ART.Conclusion CD8+Tscm are responsible for maintaining the homeostasis of other CD8+T cell subsets.CD8+Tscm play an important role in inhibiting viral replication.

Objective To study the effect of misoprostol used to intenerate cervix and ease pain in hys- teroscopy.Methods 380 cases were divided into two groups randomly;260 cases of them were placed misoprostol 200?g in vagina 2 hours before hysteroscopy and 120 cases treated without drugs served as control.The diameter of cerical,hemorrhage,the rate of PAAS and bellyache were observed.Results The diameter of cerical,hemorrhage. the rare of PAAS and bellyache in the group of misoprostol were lower than those in the group of antitheses(P

OBJECTIVETo analyze the inoculation ratio and echinacoside content of Cistanche tubulosa and provide theoretical basis for Tamarix introduction, resource protection and screening of C. tubulosa.

METHOD8 Tamarix species were introduced in the North China Plain and inoculation of C. tubulosa was conducted on all species. Phenylethanoid glycosides fingerprinting and echinacoside content of C. tubulosa were analyzed by using HPLC.

RESULTThe adaptability of 8 Tamarix species were significantly different, phenylethanoid glycosides component of C. tubulosa on T. gansuensis and T. austromongolica were basically identical in contrast to T. chinensis, echinacoside content showed no obvious difference in C. tubulosa plant growing 4 months.

CONCLUSIONT. gansuensis and T. Austromongolica are suitable for the host introduction plant of C. tubulosa resource protection and screening in North China Plain.

China ; Cistanche ; chemistry ; growth & development ; Conservation of Natural Resources ; Ecosystem ; Glycosides ; analysis ; Phenols ; analysis ; Plants, Medicinal ; chemistry ; growth & development ; Rain ; Soil ; Tamaricaceae ; classification ; growth & development

Spiramycin (SP) belongs to the 16-member macrolide antibiotics. It contains three components,namely SP I, SP II and SP III, which differ structurally in the acylation moieties on the C3 of the lactone. The SP I component contains a hydroxyl group at C3. SP II, and SP III are formed by further acetylation or propionylation of the C3 of SP I, by the same 3-O-acyltransferase (3-O-AT) . The study focused on simplifying spiramycin components. Theoretically, disruption/deletion of the 3-O-AT gene will reduce/stop the acylation of SP I to SP II and SP III. In this study, degenerated primers were designed according to the conserved regions of 3-O-acyltransferase, MdmB and AcyA in the medicamycin and carbomycin producers of S. mycarofaciens and S. thermotolerans, respectively, and an 878bp DNA fragment was amplified from the spiramycin-producer of S. spiramyceticus F21. Blast analysis of the 878bp DNA fragment suggested that it encoded the 3-O-acyltransferase (3-0-AT, sspA) gene for spiramycin biosynthesis. The flanking regions of this 878bp DNA fragment were then amplified by single-oligonucleotide-nested PCR, and a total of 4.3 kb DNA was obtained (3457nt among the 4.3kb fragment was sequenced, and deposited in GenBank DQ642742),covering the whole putative 3-O-acyltransferase gene, sspA. The sspA was then deleted from the S. spiramyceticus F21 genome by double cross-over homologous recombination, mediated by temperature-sensitive plasmid pKC1139. A comparison was done of the components of spiramycins produced by the sspA-deleted mutant strain with that of the parent strain by HPLC analysis, which showed that sspA-deleted mutant produced SP I (72%), SP II (18%), and SP III (9.6%), whereas parent strain produced SP I (7.8%), SP II (67%), and SP III (25%), respectively, demonstrating the role of ssp A in the acylation of SP I into SP II and SP III. The ssp A-deleted mutant strain obtained in this study may be used for the production of SP I, or may serve as a good starter for the construction of spiramycin derivatives.
Acyltransferases ; genetics ; Aminoglycosides ; biosynthesis ; Gene Deletion ; Genes, Bacterial ; genetics ; Genetic Engineering ; methods ; Streptomyces ; enzymology ; genetics

This study aimed to understand the dynamic distribution of infectious bronchitis virus (IBV) Jin-13 strain in SPF chickens. Ninety-day-old SPF chickens were inoculated with Jin-13, a virulent strain, and dissected at day 1, 4, 7, 10, 14, 21, 28 or 35 post-inoculation (dpi). Samples of heart, liver, spleen, lung, trachea, kidney and duodenum were collected and the N gene was detected by Sybr Green I real-time quantitative RT-PCR assays. The established method had a good linear correlation from 7.77 x 10(8) to 10(0) copies/microL. SPF chickens developed typical clinical signs of IBV at the 4th dpi, and the IBV viral concentration of tissues and organs gradually increased with a peak of up to 7.13 x 10(4) copies/microL. The viral concentration of most organs decreased by the 10th dpi, but those of the kidney, trachea and lung remained positive for IBV at 28 dpi and the heart was still positive for IBV at > 35 dpi. The results of this study, showed that the Jin-13 strain can cause prolonged virus excertion in chickens with severe renal damage.
Animals ; Chickens ; Coronavirus Infections ; veterinary ; virology ; Infectious bronchitis virus ; isolation & purification ; pathogenicity ; physiology ; Lung ; virology ; Poultry Diseases ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; Specific Pathogen-Free Organisms ; Trachea ; virology ; Virulence

Objective: To understand the epidemiological characteristics of human infection with avian influenza A (H7N9) virus in China, and provide evidence for the prevention and control of human infection with H7N9 virus. Methods: The published incidence data of human infection with H7N9 virus in China from March 2013 to April 2017 were collected. Excel 2007 software was used to perform the analysis. The characteristics of distribution of the disease, exposure history, cluster of the disease were described. Results: By the end of April 2017, a total of 1 416 cases of human infection with H7N9 virus were confirmed in China, including 559 deaths, the case fatality rate was 39.5%. In 2016, the case number was lowest (127 cases), with the highest fatality rate (57.5%). The first three provinces with high case numbers were Zhejiang, Guangdong and Jiangsu. The median age of the cases was 55 years and the male to female ratio was 2.3∶1. Up to 66% of cases had clear live poultry exposure history before illness onset, 31% of cases had unknown exposure history and only 3% of the cases had no live poultry exposure history. There were 35 household clusters (5 in 2013, 9 in 2014, 6 in 2015, 5 in 2016, 10 in 2017), which involved 72 cases, accounting for 5% of the total cases. Conclusions: The epidemic of human infection with H7N9 virus in China during 2013-2017 had obvious seasonality and spatial distribution. There was limited family clustering. Infection cases were mostly related to poultry contact.
Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Animals ; China/epidemiology* ; Cluster Analysis ; Disease Outbreaks ; Epidemics ; Female ; Humans ; Incidence ; Influenza A Virus, H7N9 Subtype/isolation & purification* ; Influenza, Human/virology* ; Male ; Middle Aged ; Poultry ; Sex Distribution

OBJECTIVETo investigate the relationship between polymorphisms of the methylenetetrahydrofolate reductase (MTHFR) C677T or A1298C and the response to fluoropyrimidine (5-FU)-based chemotherapy in advanced stomach cancer (SC).

METHODS75 cases with advanced SC were analyzed. All patients were treated with 5-FU-based chemotherapy and DNA of peripheral blood leukocytes was obtained before therapy. MTHFR genotypes were detected by PCR-RFLP method.

RESULTS(1) Of all the cases, the frequencies of MTHFR C677T C/C, C/T and T/T genotype were 32.0%, 44.0% and 24.0%, while the frequencies of MTHFR A1298C A/A, A/C and C/C genotype were 69.3%, 29.3% and 1.3%, respectively. The overal response rate to 5-FU-based chemotherapy was 29.3%. (2) The response rate to therapy among MTHFR C677T T/T genotype patients (83.3%) was significantly higher than the C677T C/T genotype (15.2%, chi(2) = 22.27, P = 0.000) or the C677T C/C genotype (8.3%, chi(2) = 23.44, P = 0.000). As compared with patients with C677T C allele, patients with C677T T/T genotype had a 7.64-fold sensitivity to 5-FU-based chemotherapy (adjusted for sex, age, prior adjuvant therapy and chemotherapy program, 95% CI: 3.14 - 18.62). The response rate to therapy among patients with MTHFR A1298C A/A genotype (36.5%) was significantly higher than patients with A1298C C allele (13.0%, chi(2) = 4.19, P = 0.041, adjusted OR = 3.75, 95% CI: 0.94 - 14.87). The response rate to therapy among patients with MTHFR C677T T/T and A1298C A/A genotypes (86.7%) was significantly higher than other groups of C677T and A1298C genotypes (15.0%, Fisher exact: P = 0.000, adjusted OR = 6.57, 95% CI: 2.8 - 15.6). (3) The incidence rates of nausea/vomiting in MTHFR C677T T/T, C/T or A1298C A/A genotypes were significantly higher than other genotypes, but the incidence rates of other treatment-related adverse reaction in MTHFR C677T or A1298C genotypes were not significantly different.

CONCLUSIONThese results in the present study suggested that the polymorphisms of MTHFR were associated with clinical response to 5-FU-based chemotherapy, suggesting that MTHFR genotypes could identify advanced SC patients that would be responsive to 5-FU-based chemotherapy.

Adult ; Aged ; Antimetabolites, Antineoplastic ; therapeutic use ; Drug Resistance, Neoplasm ; genetics ; Female ; Fluorouracil ; therapeutic use ; Genotype ; Humans ; Male ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Stomach Neoplasms ; drug therapy ; enzymology

OBJECTIVETo analyze the gene expression level of fibroblast activation protein in HBV related hepatocellular carcinoma patients and discuss its clinical significance.

METHODSFAP gene expression in 33 hepatocellular carcinoma patients cancer tissues, peficancerous tissues, distant relative normal liver tissues and 13 normal liver tissues were examined by reverse transcription PCR; and real-time fluorescent quantitative PCR (qRT-PCR) was used to quantify their expression.

RESULTSFAP were expressed in all the tissues,the relative expression values in cancer tissues, peficancerous tissues and distant relative normal liver tissues were 5.14 +/- 6.69, 1.58 +/- 0.96, 1.63 +/- 0.94, respectively, the differences were statistically significant (F = 4.401, P < 0.05); and in TNM stage I, II, IIII, they were 2.89 +/- 3.35, 4.15 +/- 4.69, 10.09 +/- 9.51 respectively; in well-differentiated, differentiated and poorly differentiated hepatocellular carcinoma were 1.62 +/- 1.74, 3.84 +/- 3.79, 1.26 +/- 13.34 respectively. The differences were all statistically significant (P < 0.05).

CONCLUSIONFAP may play an important role in the occurrence and development of HBV related hepatocellular carcinoma.

Adult ; Aged ; Carcinoma, Hepatocellular ; etiology ; metabolism ; Female ; Gelatinases ; genetics ; physiology ; Gene Expression Regulation, Neoplastic ; Hepatitis B ; complications ; Humans ; Liver Neoplasms ; etiology ; metabolism ; Male ; Membrane Proteins ; genetics ; physiology ; Middle Aged ; RNA, Messenger ; analysis ; Serine Endopeptidases ; genetics ; physiology

OBJECTIVETo establish a rapid and simple method to detect Y chromosome microdeletions directly using whole blood as starting material for multiplex-PCR.

METHODSUsing a self-prepared pHpH-Bufferq, multiplex-PCR amplification was directly carried out from the anticoagulant whole blood sample without DNA extraction step. Twelve sequence tagged sites (STS), namely SY84, SY86, SY127, SY134, SY124, SY132, SY152, SY157, SY239, SY242, SY254 and SY255, in AZFa, AZFb, and AZFc gene regions were detected in 5 different tubes. In order to ensure the validity of the experiments, sex-determining region Y (SRY) and X-linked or Y-linked zinc finger gene (ZFX/Y) were used as internal controls. Furthermore, conventional PCR using genomic DNA extracted from each blood sample was performed in parallel for evaluating the accuracy of the experiments.

RESULTSA total of 156 male samples were detected, and a normal male sample and a female sample were used as a positive and a negative control respectively. The results showed that 144 samples had no deletion; one sample was AZF-deleted; one sample was AZFb-deleted; seven samples were AZFc-deleted; one sample was both AZFb- and AZFc- deleted; and two samples were all AZFa-, AZFb- and AZFc- deleted. The observed results from two kinds of starting material (whole blood and purified DNA) are completely consistent.

CONCLUSIONIn our method, PCR amplification was directly carried out from whole blood without any DNA extraction step. So it has the advantages of low cost, simple process, time-saving operation and less cross-contamination. The whole process can be completed within 2 hours. Thus the efficiency of clinical detection is improved greatly.

Azoospermia ; genetics ; Cells, Cultured ; Chromosome Deletion ; Chromosomes, Human, Y ; Female ; Humans ; Male ; Oligospermia ; genetics ; Polymerase Chain Reaction ; Sex Chromosome Aberrations

OBJECTIVETo demonstrate the effect of bromoxynil on membrane potential and respiratory control rate (RCR) in isolate mitochondria from mice liver tissue in vitro and the intervention of NAC.

METHODSThe mitochondrial was randomized to control group, bromoxynil-poisoned group and NAC-protected group. S3, S4 and RCR of the mitochondria in each sample was detected by the method of oxygen electrode. Each sample was stained by JC-1 and the changes of membrane potential of mitochondria were observed under fluorescence microscope.

RESULTSThe S3 [(0.031 +/- 0.008) nano atoms oxygen x mg(-1) x min(-1)], RCR (1.820 +/- 0.181) of bromoxynil-poisoned group and RCR (4.253 +/- 0.210) of NAC-protected group were significantly lower than those of control group (P<0.01); the S4 [(0.017 +/- 0.004) nano atoms oxygen x mg(-1) x min(-1)] of NAC-protected group was significantly higher than control group (P<0.01). The S3 [(0.046 +/- 0.005) nano atoms oxygen x mg(-1) x min(-1)] and RCR of NAC-protected group were significantly higher than group B (P<0.01), S4 [(0.011 +/- 0.001) nano atoms oxygen x mg(-1) x min(-1)] of NAC-protected group was significantly lower than bromoxynil-poisoned group (P< 0.01). Observation under fluorescence microscope: the red fluorescence of mitochondria was dim or disappeared in bromoxynil-poisoned group while brightened in NAC-protected group but still dimmer than control group.

CONCLUSIONIn vitro, the mitochondrial RCR and the mitochondrial membrane potential are decreased after the mitochondria is incubated with bromoxynil, and NAC could improve it.

Acetylcysteine ; pharmacology ; Animals ; Electron Transport ; drug effects ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Mice ; Mice, Inbred ICR ; Mitochondria, Liver ; drug effects ; metabolism ; Nitriles ; toxicity