Adolescent ; Adult ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Female ; Flavonoids ; therapeutic use ; Hepatitis B, Chronic ; drug therapy ; Humans ; Male ; Middle Aged ; Phytotherapy ; Salvia miltiorrhiza

Objective To investigate a potential role for ORF50 promoter of human herpesvirus 8(HHV 8)in reactivation of HHV 8 in primary effusion lymphoma(PEL)BC 3 cells by HIV 1 related inflammatory cytokine,hepatocyte growth factor/scatter factor(HGF/SF). Methods The persistent stimulation of BC 3 was performed by recombinant HGF/SF and treated BC 3 cells were collected at the 3 rd and 7 th day after persistent stimulation respectively. Northern blot, quantitative PCR(real time PCR), and electron microscopy(EM)were carried out to detect the expression of mRNA of minor capsid protein ORF26 and the presence of viral particles of HHV 8 from treated BC 3 cells. Co transfection of BC 3 and human umbilical veil endothelial cells (HUVECs) with HHV 8 ORF50 promoter driven luciferase construct built previously by us and recombinant expression plasmid named pEV containing HIV 1 Tat coding gene was performed and stimulation of co transfected cells with recombinant HGF/SF and/or HIV 1 recombinant gp120 was conducted at 24 hours after transfection to induce luciferase gene expression. Then relative luciferase activity unit(RLU) was calculated at 24 hours after stimulation. In the meantime, stimulation of co transfected cells with 12 O tetradecanoylphorbol 13 acetate(TPA) was performed to be used as positive control. Results It was found that HGF/SF induced an 4.1 folds increase in mRNA expression of ORF26 when it was added individually to BC 3 cells at the 7 th day after persistent stimulation. Viral particles of HHV 8 were readily identified in BC 3 cells stimulated with HGF/SF at the 7 th day with EM analysis. Recombinant HGF/SF could upregulate promoter activity of HHV 8 ORF50 promoter in BC 3 and HUVECs cells and however, HIV 1 Tat and gp120 failed to act synergistically with HGF/SF to enhance ORF50 promoter activity in HUVECs. Conclusions The results show that HIV 1 related inflammatory cytokine HGF/SF is responsible for the induction of HHV 8 lytic cycle replication and the induction may be, at least partly mediated by ORF50 promoter of HHV 8.

Objective To observe the proliferation and phenotype-switching of pulmonary arterial smooth muscle cell (PASMC) induced by hypoxia and interfered by Ad-PKGIα. And to investigate the potential regulative role of PKGIα gene in the molecule mechanism of hypoxia pulmonary vessel remodeling (HPVR). Methods To establish the pure PASMC cultured by tissue-sticking methods. Semi-quantitative reverse transcription and polymerase chain reaction (RT-PCR) and Western blot were used to examine the PKGIα mRNA and protein expression after PASMC were transfected by Ad-PKG. The mRNA and protein expressive change of smooth muscle α actin(SM-α-actin) determined the degree of cell phenotype-switching. The changes of PASMC proliferation were determined by flow cytometry and ~3H-TdR incorporated way. Results Ad-PKGIα could transfect into PASMC and highly express. Hypoxia down-regulated the expression of SM-α-actin protein (44. 25±5.34 in normoxia, 32. 18±4. 19 in 12 h hypoxia condition, 21.90 ±2. 44 in 24 h hypoxia condition, P < 0. 05), that could be blocked by the transfeetion of Ad-PKGIα. Hypoxia could push PASMC mitosis and proliferating(~3H-TdR incorporated way: 7570 ± 371 in normoxia,12 020± 831 in 12 h hypoxia condition,14 924 ± 1491 in 24 h hypoxia condition, P <0. 05), that could be blocked by the transfection of Ad-PKGIα, too. Conclusions The results suggested that PKGIα signaling pathway might play an important role in the molecule mechanism of HPVR. And PKGIα gene might be a target point of gene therapy.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Azithromycin ; pharmacology ; Bacteremia ; microbiology ; Child ; Child, Preschool ; Culture Media ; Drug Resistance, Bacterial ; Drug Resistance, Multiple, Bacterial ; Escherichia coli ; drug effects ; isolation & purification ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Methicillin Resistance ; Middle Aged ; Penicillin G ; pharmacology ; Salmonella paratyphi A ; drug effects ; isolation & purification ; Staphylococcus aureus ; drug effects ; isolation & purification ; Staphylococcus epidermidis ; drug effects ; isolation & purification

Objective:To investigate the association between LPHN3 andattention-deficit/hyperactivity disor-der(ADHD)in Chinese Han children.Methods:Based on the Diagnostic and Statistical Manual of Mental Disor-ders,Fourth Edition (DSM-IV)diagnosis criteria,921 normal controls and 1052 ADHD children were included in the study.The Clinical Diagnostic Interview Scale (CDIS )was used to assess symptoms and ADHD sub-types.ADHD was divided into three subtypes,namely ADHD inattentive type (ADHD-I),ADHD hyperactive-im-pulsive type (ADHD-HI),and ADHD combined type (ADHD-C).The ADHD rating-scale was used to assess ADHD symptoms.Including inattentive symptoms,hyperactive symptoms,impulsive symptoms and ADHD total symptoms.Three single nucleotide polymorphisms (SNPs)of LPHN3 were genotyped.Case-control studies were conducted to investigate the association of each SNP with the ADHD and the subgroups using chi-square test.Results:rs11131347 was associated with ADHD P <0.05,OR =0.86(0.76 -0.98)],but the difference didn't survive significance after corrections,The frequency of minor allele C in cases and control was0.409 vs.0.445.For different genders,rs11131347 was significantly associated with ADHD boys [P <0.05,OR =0.82(0.71 -0.96)], The frequency of minor allele C in cases and control was0.402 vs.0.449.For different subtypes,rs11131347 was associated with ADHD-C [P <0.05,OR =0.85(0.74 -0.98)]and ADHD-C boys[P <0.05,OR =0.82(0.70 -0.97)],but none of the difference survived significance after corrections,The frequencies of minor allele C in cases and control were respectively 0.407 vs.0.445 and 0.401 vs.0.449.Genotypes distribution analysis indicated that rs11131347 was associated with ADHD in general (recessive model,P <0.05),ADHD boys (additive model,P <0.05;dominant model,P <0.05;recessive model,P <0.05),ADHD-C(recessive model,P <0.05)and ADHD-C boys (dominant model,P <0.05 ),however,none of the difference survived significance after correc-tions.rs11131347 was nominal associated with impulsive scores(P <0.05).Conclusion:These findings suggest that the polymorphism of LPHN3 is probably involved in the pathological mechanisms of ADHD and its core symptoms of impulsivity.

Objective To investigate the association of uncoupling protein 2 ( UCP-2 ) gene promoter -866G＞A polymorphism and ischemic stroke in diabetic patients.Methods A total of 844 type 2 diabetic patients including 404 cases with ischemic stroke and 440 cases without ischemic stroke were selected for the 4 year prospective study,Genomic DNA was extracted from the whole blood samples of subjects,UCP-2 gene promoter -866G ＞ A polymorphism was detected by TaqMan MGB probe method,and then the genotype and allele gene frequencies were compared.Results The risk of ischemic stroke in type 2 diabetic female patients with AA+GA genotypes of UCP-2 was higher than that with GG genotype (P＜0.05),but there was no difference among male patients with three genotypes.Conclusions UCP-2 gene promoter -866G ＞ A polymorphism increases the risk of ischemic stroke in Chinese diabetic women.

Purpose To investigate the effect of tolerogenic dendritic cells ( DC) on T lymphocytes in the spleen during the develop-ment of zymosan-induced sepsis in mice, and to explore whether PD-L1 blockade could alleviate the immunosuppressive effect of tolero-genic DC on T lymphocytes. Methods Mice sepsis model was established by intraperitoneal injection of zymosan. Splenic DC and T lymphocytes were isolated respectively by using anti-CD11c and anti-CD3 magnetic beads. The expressions of PD-L1, PD-1 and PIR-B on splenic DC were measured, and IL-12 and IL-10 secreted from DC were determined. Mitogen-induced T lymphocyte proliferation and IL-2 secretion were assessed. Anti- PD-L1 antibody was added into mixed culture of tolerogenic DCs with normal Tcells. T cell proliferation and IL-2, IL-12 and IL-10 concentrations in the supernatant of mixed culture were determined. Results At 5 days and 12 days after zymosan injection, the expressions of PD-L1, PD-1 and PIR-B on splenic DC increased greatly, secretion of IL-12p70 re-duced and that of IL-12p40 and IL-10 augmented in DC, which were associated with decrease of T cells proliferation and IL-2 secre-tion. Administrating anti-PD-L1 antibody into the mixed culture of tolerogenic DC and Tcell could alleviate the suppression of DC on T lymphocyte proliferation and secretion of IL-2, and ameliorate the ability of DC secreting IL-12 and IL-10 as well. Conclusions At late stage of zymosan-induced sepsis, the formation of splenic tolerogenic DC resulted in immunosuppression of T lymphocytes. Anti-PD-L1 antibody could improve the immunoactivity of DC and T lymphocyte through intervening PD-L1/PD-1 pathway.

Aim To observe the influences of ginsenoside Rg1 on the spontaneous contraction of small intestine smooth muscle of rabbits in vitro and explore the mechanism.Methods With the isothermal perfusion of small intestine in vitro,the influences of ginsenoside Rg1 on the spontaneous contraction of small intestine was observed and the mechanism of ginsenoside Rg1 was studied.Results Ginsenoside Rg1 reduced the amplitude of contraction of small intestine smooth muscle in rabbits in a dose-depended manner.Bay K8644 and nitro-L-arginine methylester(L-NAME)could completely block the inhibition of ginsenoside Rg1 on the contraction of small intestine smooth muscle.Ginsenoside Rg1 inhibited the intracellular calcium-depended contraction induced by rynodine in the Ca2+ free Tyrode's solution.Conclusions Ginsenoside Rg1 inhibits the contraction of small intestine smooth muscle of rabbits in vitro.The mechanism may be related to increase NO concentration in small intestine smooth muscle so that it inhibits extracellular Ca2+ inflowing via cell membrane and intracellular Ca2+ releasing via sarcoplasmic reticulum.

BACKGROUND: von Willebrand factor (vWF) is only released from endothelial cells and platelets and is an in vivo and in vitro marker of endothelial injury in septic patients with acute lung injury (ALI). Interleukin-8 (IL-8), as a proinflammatory mediator causing recruitment of inflammatory cells, induces an increase in oxidant stress mediators and makes it as a key parameter for localized inflammation. However, it has not been well established whether the level of serum IL-8 is associated with the severity of lung injury and whether it is a prognosis marker for severe lung contusion. This study was to investigate the expression of plasma vWF and IL-8 and their association with the severity and outcomes of severe pulmonary contusion. METHODS: A total of 63 patients were divided into a severe pulmonary contusion with acute respiratory distress syndrome (ARDS) group and a non-ARDS group, or a survivor group and a non-survivor group, or an injury severity score (ISS) <20 group and an ISS ≥20 group. Another 20 healthy volunteers served as controls. The levels of plasma vWF and serum IL-8 were measured by enzyme-linked immunosorbent assay (ELISA) at 1, 3, 5 and 7 days after injury. The expression patterns of the plasma vWF and serum IL-8 were compared between different groups. RESULTS: The concentrations of plasma vWF and serum IL-8 were significantly increased in all severe pulmonary contusion patients at all time points in comparison with the control group. The concentrations of plasma vWF in patients with ARDS increased during the whole study period, but vWF in patients with non-ARDS increased gradually until day 5 and then decreased at day 7. The concentration of serum IL-8 showed a similar expression pattern in both groups, but the expression increased more significantly in the ARDS group than in the non-ARDS group. Interestingly, both plasma vWF and serum IL-8 levels steadily increased in the non-survivor group. Furthermore, the level of plasma vWF was higher in the ISS≥20 group than in the ISS<20 group. The level of serum IL-8 in the ISS≥20 group was consistently high, while that in the ISS<20 group peaked at day 3 and decreased at day 5. In addition, the level of plasma vWF was positively correlated with platelet count, but negatively correlated with oxygen index. The level of serum IL-8 was positively correlated with white blood cell count and ISS score, and inversely correlated with oxygen index. CONCLUSION: The elevated levels of plasma vWF and serum IL-8 in severe pulmonary contusion patients reflect the severity of pulmonary injury and patients outcomes, suggesting that the plasma vWF and serum IL-8 are sensitive markers for clinical evaluation of the severity of pulmonary injury and predication of patient prognosis.

Artemisia lactiflora is an important medicinal plant in China. The antitumor and antioxidant activities of the extracts of 54 endophytic fungi from the plant were screened via MTT assay and DPPH scavenging radical assay, respectively. The bioactive strains were identified based on similarity of 5.8S gene and internal transcribed spacer (ITS) sequences. The results showed that extracts from ten (18.5%) isolates exhibited antitumor activity, and which from two (3.7%) isolates exhibited antioxidant activity. The Alternaria sp. GYBH47 strain was simultaneously having antagonistic activity against HL-60 leukemia, MCF-7 breast and COLO205 colon cell lines, and Phomopsis sp. GYBH42 strain having cytotoxic and antioxidant activities. The results indicated that endophytic fungi from Artemisia lactiflora are potential resources to find valuable bioactive components.
Antineoplastic Agents ; chemistry ; pharmacology ; Artemisia ; microbiology ; Biphenyl Compounds ; metabolism ; Cell Line, Tumor ; Endophytes ; chemistry ; classification ; physiology ; Free Radical Scavengers ; chemistry ; pharmacology ; Fungi ; classification ; physiology ; Humans ; Picrates ; metabolism