Objective To explore the distribution of parvovirus B19 (HPVB19) infection in patients with leukopenia.Methods Patients who visited the Affiliated Hospital of Hangzhou Normal University from January 2015 to June 2016 were analyzed.Patients with peripheral leucocytes count less than 3.5×109/L were included in experiment group and healthy people were included in control group.HPVB19 IgG and IgM were detected by enzyme-linked immunosorbent assay, and HPVB19 DNA was detected by quantitative polymerase chain reaction.Differences in continuous data between two groups were compared with two-sample t test and those in categorical data were compared with Chi-square test.Results A total of 79 patients were included in experiment group, including 32 males and 47 females.Ages ranged from 24 to 62 years old.And 126 healthy individuals were included in control group, including 55 males and 71 females.Ages ranged from 28 to 67 years old.The positive rates of HPVB19 IgG, IgM and DNA in experiment group were 34.2%, 5.1% and 3.8%, respectively, while those in control group were 36.5%, 0 and 0, respectively.The detection rates of HPVB19 IgM and DNA between two groups were significantly different (χ2=6.507, P=0.011 and χ2=4.856, P=0.028, respectively).Sequence analysis for 3 of the HPVB19 DNA positive samples showed that there were two single nucleotide polymorphisms in VP1/VP2 sequence from one patient, which contributed to the 153rd (L/H) and 219th (N/Y) amino acids mutations, respectively.Phylogenetic analysis found that two strains belong to genotype 1a and one strain belongs to genotype 1b.Conclusions Detection rate of parvovirus HPVB19 infection (positive rates of HPVB19 IgM and DNA) in leukopenic patients is significantly higher than healthy controls.HPVB19 should be detected before considering transfusion in leukopenic patients in clinical practice.

Objective: To explore the impact of arsenic on cholesterol efflux and the expression of ATP-binding cassette, sub-family A, member 1 ( ABCA1 ), ATP-binding cassette transporter G1 ( ABCG1 ), and scavenger receptor class B member I ( SRBI ) in macrophages, so as to provide the evidence for the mechanism of arsenic induced atherosclerosis. Methods: The human myeloid leukemia mononuclear cells ( THP-1 ), induced by phorbol myristate acetate, and mouse primary macrophages were treated with 0, 0.625, 1.25, 2.5 and 5 μmol/L NaAsO2 for 48 hours. Then the cells treated with 2.5 μmol/L NaAsO2 were changed to arsenic free mediums for 48 hours and collected every 12 hours to analyze the time effect of arsenic. The expression levels of ABCA1, ABCG1 and SRBI were determined by quantitative polymerase chain reaction and western blotting. Cholesterol efflux rates were measured by 3H isotope tracer. Results: Arsenic significantly down-regulated the expression levels of ABCA1 and ABCG1, and cholesterol efflux in a dose-dependent manner. The levels of ABCA1 mRNA decreased by 69% and 72%, the levels of ABCG1 mRNA decreased by 42% and 34%, and the rate of cholesterol efflux decreased by 55% and 59% in THP-1 and mouse primary macrophages cells treated with 5 μmol/L NaAsO2 ( all P<0.05 ). Arsenic had no significant effect on SRBI expression ( all P>0.05 ). Arsenic inhibited ABCA1 expression and cholesterol efflux in THP-1 in a time-dependent manner. Compared with cells before the exposure of arsenic, the level of ABCA1 mRNA and the rate of cholesterol efflux in THP-1 bottomed at 48 hours by 43% and 42%, and gradually recovered when arsenic was removed. Conclusions Arsenic inhibits cholesterol efflux by down-regulating the expression of ABCA1 and ABCG1 in macrophages.

Objective:To study the effects of andrographolide on the expression of IL-18 related cytokines by peripheral blood monocytes.Methods:After treatment of andrographolide in different concentrations on LPS stimulated human peripheral blood mononuclear cells (PBMC) and LPS+IFN-γ/IL-4 activated magnetic bead-sorted monocytes (PBM),the transcription level of IL-18,IL-18BP,IL-18Rα and IL-18Rβ was detected by real-time RT-PCR;and secreted IL-18,IL-18BP and IL-1β,IL-1Rα by PBM was detected with ELISA.Results:Andrographolide regulated the transcription of IL-18 and IL-18BP in a dose-and time-dependent effect.Andrographolide up-regulated the transcription of IL-18BP in LPS stimulated PBMC,and increased the ratio of IL-18BP/IL-18.The ratio of IL-18BP/IL-18 rose from 9.60 to 214 in LPS+IFNγ activated PBM,and IL-1Rα/IL-1β declined from 9 200 to 6 520 in IL-4 activated PBM by andrographolide treatment.Conclusion:Andrographolide can regulate IL-18 related gene transcription and expression in activated peripheral blood monocytes,and inhibit the excessive expression of IL-18 during inflammation.

Objective To investigate the effects of andrographolide on the expressions of Th1 cytokine [interferon (IFN)-γ] mRNA and Th2 cytokines [interleukin (ID-4, IL-10] mRNA of peripheral blood mononuclear cells (PBMCs) from patients with chronic hepatitis B (CHB) and its anti-hepatitis B virus (HBV) activity. Methods PBMCs from CHB patients were cultured with 10 mg/L andrographolide (experimental group) or 0.1% DMSO (control group). HepG2. 2. 15 cells were stimulated with andrographolide of different concentrations (experimental group) or adefovir (control group). The expressions of IFN-γ mRNA, IL-4 mRNA and IL-10 mRNA of PBMCs and the replication of HBV DNA in HepG2. 2. 15 cell line were detected by fluorescence quantitative PCR. Results The expression of IFN-γ mRNA in PBMCs cultured with 10 mg/L andrographolide for 16 h was higher than that in control group (Z=-2. 78, P=0. 05), and the expressions of IL-4 and IL-10mRNA in experimental group were lower than control group (Z= -3. 82, P<0. 01), while the ratio of IFN-γ/IL-4 mRNA was higher than control group (Z= - 3. 82, P<0. 01). Andrographolide with different concentrations had no effect on the replication of HBV DNA in HepG2. 2.15 cells ((=11. 88, P>0.05). However, adefovir had inhibitory effect on the replication of HBV DNA (t =15. 95,P< 0. 05). Conclusion Andrographolide can regulate the expressions of IFN-γ mRNA, IL-4 mRNA and IL-10 mRNA in PBMCs from CHB patients and improve Thl/Th2 balance, while it has no effect on the replication of HBV DNA.

Objective To investigate the association of the LIPC-C480T (rs1800588) and lipid levels and dyslipidemia in different age-and-sex groups in Han Chinese population.Methods The serum TC,TG,LDL-C and HDL-C were detected by automatic biochemical analyzer in 2420 health adults (1527 men and 893 women).The genotypes of rs1800588 were detected by M ALDI-TOF MS.According to the age difference (≤44,45-59 and ≥60-year-old),the total samples were divided to young (241 men and 201 women),middle-aged (652 men and 360 women) and older (634 men and 332 women) groups.The effects of genotypes on 4 serum lipid indicators in each age-and-gender group were analyzed by one way analysis of variance (ANOVA),and the odd risk of genotypes on dyslipidemia was estimated by binary Logistic regression analysis.The P value less than 0.05 was considered statistically significant.Results The frequence of allele T for LIPC rs1800588 in this population is 39.4％.In each age group the lipid parameters are quite different between males and females.Compared with those with CC genotype,middle-aged and elder men with CT or TT genotype have higher TC and HDL-C levels,and elder men with TT genotype also have higher TC level ; young women bearing CT genotype have higher TC level,and the CT and TT genotypes have higher HDL-C levels,middle aged women with CT or TT genotype have higher TC and TG levels,and CT genotype also have higher HDL-C level,the elder women with TT genotype have higher HDL-C level.Compared with those CC genotype individuals,the risk for mixed hyperlipidemia and hypercholesterolemia increases 2.318 folds (P =0.004) and 2.571 folds (P ＜ 0.001) respectively,while the risk for low HDL-C decreases 1.908 folds (P =0.029) for TT genotypes individuals among elder males; the hypercholesterolemia risk increasc 1.688 (P =0.036) and 2.099 times (P =0.040) in CT and TT genotypes respectively,and the risks for hypertriglyceridemia and mixed hyperlipidemia are 2.060 (P =0.038) and 2.381 (P =0.019) times higher than those with CC genotype among middle-aged females.Conclusions The LIPC rs1800588 site associates with the lipid levels and dyslipidmia risk in Han Chinese in an age-and-sex model.This SNP site has higher impact on lipid levels and dyslipidemia among elder males and middle-aged females,and the T allele is the risk factor.

Objective: To investigate protective effects of nicotinamide mononucleotide (NMN) on ethanol-induced DNA damage in L02 cells, so as to provide the evidence for adjuvant therapy of NMN on alcoholic liver diseases. Methods: L02 cells were pretreated with different concentrations of NMN (0, 1, 2, 4 and 8 mmol/L) for 6 h, and then were exposed to 0.4% ethanol for 12 h. The treated cells were divided into the control group, 0.4% ethanol group and different concentrations of NMN groups. Cell viability was analyzed using trypan blue staining for determining the concentration of NMN as a protective agent. The effects of NMN on ethanol-induced DNA damage in L02 cells were evaluated using immunofluorescence detection and reactive oxygen species (ROS) assay. L02 cells were exposed to 0.4% ethanol for 12 h, cultured in a medium containing a protective concentration of NMN, and divided into PBS group and NMN group. Cell viability was detected at 0, 2, 4, 8, 16 and 32 h, and the effects of NMN on repairing ethanol-induced DNA damage were evaluated by alkaline comet assay. Results: The cell viability was lower in 0.4% ethanol group than than in the control group, and was higher in different concentrations of NMN groups than in 0.4% ethanol group (all P<0.05), with no significant difference in the cells viability between 4 mmol/L and higher concentrations of NMN groups and the control group (all P>0.05). Therefore, 4 mmol/L NMN was selected as a protective agent. The cell tail moments, relative immunofluorescence intensities of γH2AX and relative levels of ROS were higher in 0.4% ethanol group than in the control group, and lower in 4 mmol/L and higher concentrations of NMN groups than in 0.4% ethanol group (all P<0.05). The cell viability was increased and the cell tail moment was shortened with the increase of 4 mmol/L NMN intervention time; and the cell viability in 4 h and more of NMN groups were higher, and the cell tail moment were lower than that in PBS group (all P<0.05). Conclusions NMN attenuates DNA damage in a dose-dependent manner and promotes the repair of DNA damage in a time-dependent manner. NMN has a protective effect on ethanol-induced DNA damage in hepatocytes.

OBJECTIVEConstruct the gene library of apoptosis related genes in acute promyelocytic leukemia (APL) cell line NB4 cells treated by arsenic trioxide to clarify the apoptotic mechanism of NB4 cells.

METHODAPL cell line NB4 cells treated with or without arsenic trioxide for 24 hours. Total RNA was extracted and suppress subtractive hybridization (SSH) was conducted according to the manual. With the cDNA of the apoptosis cells as the tester and that of control cells as the driver, forward and reverse hybridization was performed. Differentially expressed genes were linked with pGEM-Teasy cloning vector and transformed into E. coli DH5alpha. The positive clones were screened by blue and white spot. PCR were used to amplify these genes.

RESULTThe subtractive cDNA libraries related with apoptosis of NB4 cells were successfully constructed.

CONCLUSIONThe constructed subtractive libraries are suitable for further study on the functional genes associated with apoptosis ofNB4 cells induced by arsenic trioxide.

Apoptosis ; drug effects ; genetics ; Arsenicals ; pharmacology ; Cell Line, Tumor ; DNA, Complementary ; analysis ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Gene Library ; Humans ; Leukemia, Promyelocytic, Acute ; pathology ; Nucleic Acid Hybridization ; methods ; Oligonucleotide Array Sequence Analysis ; Oxides ; pharmacology ; Sequence Analysis, DNA