Objectives To increase the safety of blood supply and to evaluate the feasibility of Nucleic Acid amplification test (NAT) of blood donations in blood bank. Methods Individual donor plasma samples serologically negative for HCV, HBV and HIV detected by ELISA were pooled according to the size of 20?50 ?l. HCV RNA and HBV DNA in pooled samples were detected by AcuGen AG 9600 AmpliSensor qualitative PCR methods. Individual donor plasma samples in positive pooled samples were further tested by PCR. Results One(0.01%)of 8805 donations was PCR for HCV RNA positive. Six (0.4%)of 1 441 donations were PCR for HBV DNA positive. The whole procedure took three days from pooling donor plasma samples to identifying the positive samples. Conclusion It is feasible to incorporate NAT into ELISA screening blood donations for HBV and HCV. NAT will further increase the safety of blood supply.

ObjectiveTo analyze the levels of matrix metalloproteinase 9 (MMP-9 ) in cerebrospinal fluid (CSF) of patients with central nervous system (CNS) infection caused by different pathogens.MethodsThe levels of MMP 9 in CSF were detected by enzyme linked immunosorbent assay (ELISA) in 10 patients with tuberculous meningitis in both acute phase and recovery phase,10 purulent meningitis,10 cryptococeal meningitis,10 viral encephalitis and 10 controls.The differences among the groups were compared by t test. The correlations between MMP-9 levels and the cell count,glucose, chloride, and protein in CSF were analyzed by Spearmanrank correlation.ResultsThe CSFMMP-9levelsingroupsoftuberculousmeningitis, purulentmeningitis,cryptoeoccal meningitis and viral encephalitis were (569.46±162.42),(182.79±99.06),(54.69±19.93) and (18.52±10.31) ng/mL,respectively,which were all significantly higher than that in controls (3.51± 1.53) ng/mL. There were significant differences between tuberculous meningitis group and other groups (t=2.925,3.041,3.237,3.454;P0.0340,0.0270,0.0080 and0.0001,respectively). Moreover,in tuberculous meningitis group,the MMP-9 level in acute phase was (569.46±162.42) ng/mL,which was significantly higher than that in recovry phase (294.30+89.06) ng/mL.The CSF level of MMP-9 in tuberculous meningitis group was positively correlated with CSF protein (r=0.509,P=0.044),negative correlated with CSF glucose (r=-0.451,P=0.008) and chloride (r=-0.637,P=0.007),but no correlation with CSF cell count (r=0.308,P=0.246). And there were no correlations in other groups. ConclusionsMMP-9 level in CSF increases significantly in patients with tuberculous meningitis and purulent meningitis,which can be used as a marker of CNS infection.Dynamic monitoring of thc CSF level of MMP-9 may be meaningful for the diagnosis and treatment.

Objective To investigate the effects of neferine on the proliferation and the P-glycoprotein(P-gp) expression of refractory/relapsed acute leukemic cells and to provide experimental evidence for further clinical use. Methods MTT and immunocytochemistry SABC methods were used respectively to observe the alteration of the proliferation and the expression of P-gp in refractory/relapsed acute leukemic cells after treating with neferine. Results The inhibition ratio on acute leukemic cells of neferine adding adriamycin(ADM) group was significantly higher than that of ADM group (P0.05). Conclusion [WTBZ]Neferine can inhibit the proliferation of refractory/relapsed acute leukemic cells, and reduce the P-gp expression of refractory/relapsed acute leukemic cells and consequently reverse multidrug resistance(MDR).

Objective To improve diagnosis of straight back syndrome.Methods 21 patients with straight back syndrome include 12 male and 9 female,and aged 15～36 years old(mean 21 years old).The standard P-A and left lateral chest film were taken and the A-P diameter and transverse diameter of the chest were measured,and the ratio between the both was calculated.Results The A-P diameter of the chest were 5.3 cm～8.5 cm(mean 6.6 cm)in male and 4.2 cm～8.1 cm(mean 6.2 cm)in female.the ratio between the A-P diameter and transverse diameter of the chest were 0.22～0.37(mean 0.34)in male,and 0.19～0.366(mean 0.31)in female.Conclusion The ratio between the A-P diameter and transverse diameter of the chest ≤0.37 is the diagnostic basis of the straight back syndrome.

Objective To identify SEN virus and analyse the sequence of its partial gene in blood donors in China.Methods Two pair primers from ORF1 of SENV genome were designed and samples from blood donors were detected for SENV DNA by nested PCR.The isolates from blood donors were cloned and sequenced.Results 6 isolates were obtained in 329 sera from blood donors.The sequence analysis showed that there were about 52%～100% homology of deduced amino acid between SENV A～H genotypes and 6 isolates from blood donors. The Phylogenetic tree analysis showed that 6 SENV isolates from blood donors belong to SENVH genotype.Conclusion There exists SENVH genotype infection among blood donors in China. SENV may be transmitted through blood transfusion.

Multidrug resistance (MDR) plays a major obstacle to successful gastric cancer chemotherapy. The purpose of this study was to investigate the MDR reversal effect and mechanisms of hyperthermia in combination with neferine (Nef) in adriamycin (ADM) resistant human SGC7901/ADM gastric cancer cells. The MDR cells were heated at 42°C and 45°C for 30 min alone or combined with 10 μg/mL Nef. The cytotoxic effect of ADM was evaluated by MTT assay. Cellular plasma membrane lipid fluidity was detected by fluorescence polarization technique. Intracellular accumulation of ADM was monitored with high performance liquid chromatography. Mdr-1 mRNA, P-glycoprotein (P-gp), γH2AX expression and γH2AX foci formation were determined by real-time PCR, Western blot and immunocytochemical staining respectively. It was found that different heating methods induced different cytotoxic effects. Water submerged hyperthermia had the strongest cytotoxicity of ADM and Nef combined with hyperthermia had a synergistic cytotoxicity of ADM in the MDR cells. The water submerged hyperthermia increased the cell membrane fluidity. Both water submerged hyperthermia and Nef increased the intracellular accumulation of ADM. The water submerged hyperthermia and Nef down-regulated the expression of mdr-1 mRNA and P-gp. The water submerged hyperthermia could damage DNA and increase the γH2AX expression of SGC7901/ADM cells. The higher temperature was, the worse effect was. Our results show that combined treatment of hyperthermia with Nef can synergistically reverse MDR in human SGC7901/ADM gastric cancer cells.

Objective To observe the changes of number and proportion of CD4+CD25+FOXP3+ cells in peripheral blood of active rheumatoid arthritis patients (RA),and explore the function of recombinant human TNFR Ⅱ-Fc on the CD4+CD25-FOXP3+ Treg cells.Methods ① Forty severe RA patients were selected,who were divided into the combined treatment group (TNFR Ⅱ-Fc+MTX) and MTX only group according to the principle of randomized,double-blind,parallel and placebo-controlled study.All patients were treated for 12 weeks.Flow cytometry was used to analyze and compare the expression ratio of CD4+CD25+FOXP3 +Treg cells of RA patients' peripheral blood.Forty healthy controls were selected for parallel comparison.② VAS,DAS28,HAQ average of the two groups at different periods were compared.Matched t test was used to examine the quantity data between the groups.Results ① The proportion of CD4+CD25+FOXP3+ cells in the peripheral blood of active rheumatoid arthritis patients was significantly lower than healthy group [ (5.4±1.4)％ vs ( 7.5±1.5 )％,P＜0.01 ].The proportion of CD4+CD25+ FOXP3+ Treg cells in combined treatment group was significantly higher [(7.0+1.2)％ vs (5.2±1.6)％,P＜0.01 ] after TNFR Ⅱ-Fc and MTX combination therapy for 12 weeks.The increase rate of CD4+CD25+FOXP3+Treg cells in combined treatment group was evidently more remarkable than MTX only group [ (7.0 ± 1.2)％ vs (5.6 ±0.7 )％,P＜0.01].② After 12 weeks treatment,the arerage scores of VAS,DAS28 and HAQ of the combined treatment group were better than MTX only group,and the difference was statistically significant (P＜0.01).Conclusion This study has shown that the healing effect of TNFR Ⅱ -Fc combined with MTX is better than MTX only.TNFR Ⅱ -Fc can restore and improve the proportion of CD4+CD25+FOXP3+ Treg cells in active RA patients,which is likely to be an important mechanism for the treatment of RA patients.

Objective To examine the distribution of systemic inflammation and risk factors of cardiovascular disease (CVD) in patients with psoriatic arthritis (PsA) by comparing with healthy controls.Methods Forty PsA patients and 44 controls were recruited into this cross-sectional study.We evaluated the disease activity and severity [erythrocyte sedimentation rate (ESR),C reactive protein(CRP) and Disease Activity Score (DAS)28],functional ability in patients with predominant axial involvement [Bath AS disease activity index (BASDAI) and Bath AS functional index (BASH)],traditional CVD risk factors and inflammation between these two groups of patients.Then,we compared risk factors for CVD between 40 consecutive PsA patients and 44 controls,adjusted for body mass index (BMI).The frequencies were compared using chi-square tests for categorical variables.Student's t-tests or Mann-Whitney U-tests were used forcontinuous variables where appropriate.Association between the traditionaland metabolic risk factors and the hs-CRP level were assessed using Spearman correlations.Finally,we also assessed the role of inflammation on the CVD risk factor by using a BMI and hs-CRP-adjusted model.Results The BMI of PsA patients was significantly higher than that of the controls.After adjusting for the BMI,PsA patients had a higher prevalence of hypertension (OR=5.615,95％CI 1.844-17.099) and diabetes mellitus (OR=10.655,95％CI 1.150-98.683) than the controls.PsA patients had significantly increased systolic and diastolic blood pressures [(SBP) and (DBP)],total cholesterol (TC)/high density lipoprotein cholesterol (HDL),insulin resistance,inflammatory markers (hsCRP,white cell count and platelet) and decreased HDL compared to the controls.As excepted,the hsCRP level [4.0 (2.1-13.9) vs 1.7 (1.3-2.2)],platelet and white cell counts were significantly increased in the PsA group reflecting underlying inflammation.Further adjustment for hsCRP level rendered the differences in the prevalence of hypertension (OR=3.544,95％CI 1.151-10.914);but the DBP,HDL and sugar levels were non-significantly different between the two groups,while the differences in other parameters were significant.Conclusion The data support the hypothesis that PsA may be associated with hypertension,obesity and dyslipidemia because of the shared inflammation pathway.

Objective To analyze the characteristics and significance of matrix metalloproteinase-9 (MMP-9) expression in the pathophysiological processes of tuberculous meningitis in mice.Methods Sixteen mice were intracerebroventricularly injected with H37RV suspension as the model group.Meanwhile,the other 16 mice were injected with 0.9％ sodium chloride solution as the control group.Thirty days later,all mice were decapitated and the brain tissue were respectively used to for Mycobacterium tuberculosis (M.tuberculosis) incubation,pathological changes observation,MMP-9 activity detection by zymography,blood-brain-barrier permeability and moisture content detection,and immunofluorescence stain of MMP-9,glial fibrillary acidic protein (GFAP) and integrin αM (OX-42).The t test was used to compare the differences between the two groups.Results Every experimental mouse was injected with (1.271±0.111) × 106 colony-forming units (cfu) M.tuberculosis.Thirty days later,the amount of M.tuberculosis in brain tissue homogenates was (4.900± 1.407) × 104 cfu/mL,and the hematoxylin and eosin staining showed dilatation of subarachnoid and ventricular and infiltration of a large number of inflammatory cells.The cumulative absorbance (A) of MMP-9 bands of brain tissue was 47 821 ± 19 932 in the model group and 10 082 ± 3 544 in the control group.The difference was statistically significant (t =3.728,P=0.010).The evans blue (EB) content of brain tissue was (11.8 ± 3.6) μg/g in model group and (4.7 ±3.4) μg/g in control group.The difference was statistically significant (t=2.887,P=0.028).The moisture of brain tissue was 0.849±0.035 in model group and 0.775±0.037 in control group.The difference was statistically significant (t=2.925,P=0.026).The immunofluorescence staining showed that the infected brain tissue expressed high degrees of MMP-9,GFAP and OX-42.And MMP-9 was overlapped with both GFAP and OX-42 obviously.Conclusions The activity of MMP-9 is significantly enhanced in brain tissue of mice suffering from tuberculous meningitis and participates in blood-brain barrier damage,tissue edema and inflammatory cells exudation.Microglia cells-astrocytes network is involved in the secretion of MMP-9.

OBJECTIVE: To analyze the mdr-1 gene promoter methylation and histone acetylation status in both MCF-7/Adr and MCF-7 cells and to preliminarily explore the epigenetic mechanism of multidrug resistance in breast cancer. METHODS: mdr-1 gene promoter methylation status of the 2 cell lines was detected by methylation-sensitive PCR. mRNA expression of DNA methyltransferases (DNMTs) and histone deacetylases (HDACs) was detected by real-time quantitative PCR. Acetylation level of histone H3 and H4 was examined by optical density assay. RESULTS: Promoter hypermethylation of mdr-1 gene was observed in MCF-7 cells whereas hypomethylation was found in MCF-7/Adr cells. Expression of DNMT1, DNMT3a, and DNMT3b mRNA in MCF-7/Adr cells significantly decreased compared with that of MCF-7 cells (P<0.05). H3 and H4 histone acetylation levels of MCF-7/Adr cells were obviously higher than those of the MCF-7 cells (P<0.01). Expression of HDAC1, HDAC2, HDAC7, and Sirtuin type 1 (SIRT1) mRNA in MCF-7/Adr cells was significantly reduced (P<0.05). CONCLUSION Hypomethylation of the promoter region of mdr-1 gene, high H3 and H4 histone acetylation, and low mRNA expression of DNMTs and HDACs may be important epigenetic factors for the development of MDR in MCF-7/ Adr cells.
ATP Binding Cassette Transporter, Subfamily B, Member 1 ; genetics ; Acetylation ; Breast Neoplasms ; pathology ; Cell Line, Tumor ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; metabolism ; DNA Methylation ; genetics ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Epigenesis, Genetic ; Female ; Histone Deacetylases ; genetics ; metabolism ; Histones ; chemistry ; Humans ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; genetics ; metabolism