Objective To investigate whether HIV-1 Nef could promote the angiogenesis and tu-morigenesis induced by KSHV vIL-6 through regulating PTEN/PI3K signaling pathway .Methods Lipo-some transfection was used to transfect cDNA of pPTEN , dominant-negative ( DN) construct of PI3K and control vector into endothelial cells , which stably express KSHV vIL-6 and HIV-1 Nef.Microtubule forma-tion assay and chicken chorioallantoic membrane ( CAM) assay were used to evaluate microtubule formation and angiogenesis , respectively .Expressions of PTEN and PI 3 K were measured by Western blot .Results Both overexpression of PTEN and inhibited expression of PI 3K suppressed the vIL-6-induced microtubule for-mation and angiogenesis in CAM mediated by Nef .Conclusion HIV-1 Nef enhances vIL-6-induced angio-genesis and tumorigenesis through regulating PTEN/PI3K signaling pathway .

AIM:To investigate the effects of angiotensin II ( Ang II) on the immune maturation and the oxi-dized low-density lipoprotein (Ox-LDL)-uptaking capacity of human monocyte-derived dendritic cells (DCs).METH-ODS:Human peripheral blood mononuclear cells were isolated by density gradient centrifugation , and the monocytes were purified by positive selection with anti-CD14 magnetic beads.After cultured with rhGM-CSF (100 μg/L) and rhIL-4 (50μg/L) for 5 d, the monocytes differentiated into immature DCs .On the 6th day of the culture, the cells were treated with various concentration levels of Ang II or pretreated with losartan .The immunophenotypic expression of HLA-DR and CD83 was analyzed by flow cytometry .The secretion levels of IL-12 and IFN-γin the culture supernatants were measured by ELISA.Furthermore, DCs were incubated with DiI-labelled Ox-LDL.The DiI-Ox-LDL-incorporated fraction was investiga-ted by flow cytometry .The mRNA expression of 3 scavenger receptors , scavenger receptor A ( SR-A) , CD36 and lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1), was examined by real-time PCR.RESULTS: Ang II induced the maturation of human monocyte-derived DCs, stimulated the expression of CD83 and HLA-DR, and promoted the secre-tion of IL-12 and IFN-γ, which were suppressed by losartan .Furthermore, Ang II increased the Ox-LDL-uptaking capacity of DCs, which was partially reduced by losartan .The incubation of DCs with Ang II enhanced the mRNA expression of LOX-1 in a dose-dependent manner , which was reduced by losartan .However, the expression of SR-A and CD36 was not changed .CONCLUSION:Ang II promotes the immune maturation of human monocyte-derived DCs and increases the up-take of Ox-LDL probably through the up-regulation of LOX-1 expression.

Objective: To observe the changes of circulating fractalkine and its receptor CX3CR1 level in patients with chronic congestive heart failure (CHF).
Methods: Our work included 2 group, CHF group, n=55 patients and Control group, n=25 healthy subjects. Plasma level of soluble fractalkine (sFKN) was measured by ELISA, CX3CR1 in peripheral blood mononuclear cell was examined by lfow cytometry method. The relationship between sFKN and NT-proBNP was studied.
Results: Compared with Control group, CHF group had increased sFKN level, P=0.004, and the patients with NYHY III, IV were more than NYHY II, and CHF group also had the higher CX3CR1 expression (14.7 ± 8.1), P<0.05. The CX3CR1 level increased accordingly with NYHY classiifcation, as the patients with NYHY II, CX3CR1 was at (25.1 ± 12.4), P=0.03 compare with Control group;with NYHY III, CX3CR1 was at (37.3 ± 11.0) , P=0.04 compared with NYHY II;with NYHY IV, CX3CR1 was at (41.7 ± 11.1), P=0.009 compared with NYHY II. The circulating sFKN level was positively related to pro-BNP level (r=0.364, P<0.01).
Conclusion: The circulating FKN l and its receptor CX3CR1 might be involved in pathogenesis of immune-inlfammatory pathogenesis in CHF patients.

Objective To assess the academic level of papers on systematic reviews and meta-analysis published in Chinese Journal of Pediatricsand methodology they used.Methods Basic data were extracted from 13 papers on sys-tematic reviews and meta-analysis published in Chinese Journal of Pediatrics .The methodology they used was assessed according to the preferred reporting items for systematic reviews and meta-analysis (PRISMA) and ANSTAR Scale and analyzed using the RevMan5.0.Results The PRISMA score was 14-23.5 (mean 20.0±3.11) and the AMSTAR score was 3-7.5 (mean 6.04±1.38) for the methodology used in papers on systematic reviews and meta-analysis published in Chinese Journal of Pediatrics .Conclusion The methodology used in papers on systematic reviews and meta-analy-sis published in Chinese Journal of Pediatrics is not quite valid and should thus be improved .

Objective To observe the dynamic changes of heme oxygenase 1,NAD(P)H:quinine oxidoreductase 1 and Nuclear factor-E2-related factor 2 in the lung tissue of acute H2S-intoxicated rats and intervention effects of ulinastratin(UTI).Methods A total of 96 SD rats of clean grade were divided randomly(random number)into four groups:normal control group(NS group,n =8),UTI control group(UTI group,n =8),H2S-intoxicated model group(H2S group,n =40,rats were exposed to H2S(200 × 10-6)for 1 h to establish the H2S-intoxicated model)and UTI treatment group(H2S +UTI group,n =40,rats were intraperitoneal injected with the dose of UTI 105 U/kg).H2S group and H2S + UTI group were sacrificed 2,6,12,24 and 48 h after modeling.The activity and mRNA expression of HO-1 and NQO-1 in the lung tissue were measured by ELISA and RT-PCR methods,and the expression of Nrf2 mRNA and protein in the lung tissue was detected by RT-PCR and Western Blot methods.Pathological changes of lung tissue were observed by lightmicroscope and the lung injury score was used to evaluate inhalation injury.Results The pulmonary HO-1 activity and mRNA expression in rats of H2S group at 2,6,12 h(P ＜ 0.01)after intoxication were markedly increased than that in NS group:In comparison with H2S group,the pulmonary HO-1 activity and mRNA expression increased at 6,12,24,48 h(P ＜0.01).The pulmonary NQO-1 activity and mRNA expression in rats of H2S group at 2,6,12,24 h(P＜ 0.01)after intoxication were markedly increased than that in NS group; In comparison with H2S group,the pulmonary NQO-1 activity and mRNA expression increased at 6,12,24,48 h(P ＜ 0.01).The pulmonary Nrf2 mRNA and protein expression in rats of H2S group at 2,6,12 h(P ＜0.01 or P ＜0.05)after modeling were markedly increased than that in NS group and reached peak 2 hour after modeling; In comparison with H2S group,the pulmonary Nrf2 mRNA and protein expression increased at 6,12,24,48 h(P ＜0.01).At 24 h after modeling,the degree of lung damage were also decreased in H2S group compared with H2S + UTI group in the lightmicroscope.Histopathological examination showed that the degree of lung injury in H2S + UTI group was less severe than that in H2S group especially in the 12,24 and 48 h (P ＜0.01).Conclusions HO-1,NQO-1 and Nrf2 are involved in the pathogenesis of acute lung injury induced by H2S-intoxicated in rats.UTI may improve the imbalance in redox and activate HO-1,NQO-1 and Nrf2 can reduce lung injury and protect the lung injury induced by H2S in rats.

ObjectiveTo investigate the protective effect of paraoxonase 1 (PON1) gene against liver oxidative stress injury in mice due to dichlorvos poisoning.Methods Experiment 1: 12 male Balb/c mice were randomly divided into three groups, with 4 mice in each group: control group, green fluorescent protein lentivirus control group (Lv-GFP group), and recombinant PON1 lentivirus group (Lv-PON1 group). 2×107 TU of Lv-GFP or Lv-PON1 was transfected via tail vein, while normal saline was given to those in control group. Blood was collected on 0, 1, 3, 5, 7, 9 days via fundus venous plexus for the assay of serum PON1 activity. PON1 mRNA and protein expression levels were respectively determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot on the 3rd post-lentivirus transfection day. Experiment 2: according to the random number table method, another 96 male Balb/c mice were divided into four groups of 24 mice in each control group, dichlorvos group, Lv-GFP intervention group, and Lv-PON1 intervention group. Lv-GFP or Lv-PON1 was transfected via tail vein followed by intraperitoneal injection of dichlorvos 9 mg/kg, while those in control group were given normal saline. Six mice in each group were sacrificed respectively at 6, 12, 24, 48 hours, and liver tissue was collected. PON1 mRNA and nuclear factor E2-related factor 2 (Nrf2) mRNA expression levels were determined by RT-PCR, and PON1 protein level was determined by Western Blot. The content of malondialdehyde (MDA) and glutathione (GSH) in the liver tissue were determined by chemical colorimetry. The activity of superoxide dismutase (SOD) and catalase (CAT) were measured by double antibody sandwich enzyme linked immunosorbent assay (ELISA).Results Experiment 1: after Lv-PON1 was transfected to normal mice, PON1 activity in serum gradually increased and maintained a high level on 3rd day, while that of the control group and Lv-GFP group showed a normal low level. On the 3rd post-lentivirus transfection day, PON1 mRNA and PON1 protein expressions in liver were significantly higher than those of control group and Lv-GFP group. Experiment 2: compared with control group, the mice in dichlorvos group showed significant decreases in PON1 mRNA, PON1 protein, Nrf2 mRNA as well as GSH, SOD, CAT levels at 6 hours [PON1 mRNA (gray value):0.237±0.075 vs. 0.674±0.011, PON1 protein (gray value): 0.602±0.086 vs. 0.998±0.124, Nrf2 mRNA (gray value): 0.089±0.012 vs. 0.126±0.010, GSH (mg/g): 3.84±0.33 vs. 5.52±0.40, SOD (μg/g): 0.383±0.040 vs. 0.564±0.052, CAT (ng/g): 7.32±1.28 vs. 12.46±1.54, allP< 0.05], and remarkable increase in MDA content (nmol/g: 7.78±0.41 vs. 2.34±0.25,P< 0.05). With the extension of time, PON1 mRNA, PON1 protein, Nrf2 mRNA and GSH, SOD, CAT levels gradually increased, MDA content gradually decreased, Nrf2 mRNA expression level had risen to the level of control group at 24 hours (0.133±0.019 vs. 0.126±0.009,P> 0.05), and it was higher than that of the control group at 48 hours (0.206±0.028 vs. 0.124±0.010,P< 0.05). Compared with that of the dichlorvos group, Lv-PON1 intervention group showed a significant increase in PON1 mRNA, PON1 protein, Nrf2 mRNA and GSH, SOD, CAT levels [PON1 mRNA (gray value): 0.726±0.021 vs. 0.237±0.075, PON1 protein (gray value): 0.739±0.050 vs. 0.602±0.086, Nrf2 mRNA (gray value): 0.158±0.007 vs. 0.089±0.012, GSH (mg/g): 4.30±0.22 vs. 3.84±0.33, SOD (μg/g): 0.454±0.062 vs. 0.383±0.040, CAT (ng/g): 8.98±1.02 vs. 7.32±1.28, allP< 0.05], and a decrease in MDA content (nmol/g: 6.56±0.44 vs. 7.78±0.41,P< 0.05).Conclusion Regulation of PON1 gene can reduce MDA content, enhance SOD and CAT activities, increase GSH content, and it may also up-regulate Nrf2 mRNA expression to play a protective effect against oxidative stress of liver injury induced by dichlorvos poisoning.

Objective To report a familial dentinogenesis imperfecta type Ⅱ (DGI type Ⅱ) with a novel splicing mutation in DSPP (dentin sialophosphoprotein) gene.Methods Based on the result of linkage analysis performed previously to map the candidate gene DSPP in the family, the promoter,the first four exons and exon-intron boundaries of DSPP were directly sequenced for the members of the DGI type Ⅱ family. Denaturing high performance liquid chromatography (DHPLC) analysis was performed to confirm the results of sequencing.Results A novel splicing mutation of 23 bp deletion in intron 2 of DSPP gene was identified by DNA sequence analysis. The mutation changed acceptor site sequence from CAG to AAG, and might result in functional abolition of possible branch point site in intron 2. DHPLC result was consistent with that of sequencing. The mutation may be identified in all affected individuals, but not found in normal members of the family and 50 controls.Conclusion These results suggest the deleted mutation of DSPP gene causes DGI type Ⅱ in the family. The mutation has not been reported before.

Objective: To report a case of azoospermia with a karyotype of 45,X,der(Y)t(Y;13)(q11.2;q12),-13,accompanied with slight bilateral gynecomastia and multiple nodules.Methods: The karyotype was identified by karyotyping and FISH,and the breakpoints of the Y chromosome and the copy number of the BRCA2 gene in 13q12 determined by PCR-STS and DNA polymorphic analysis.The testis and nodule tissues of the patient were obtained for biopsy.Results: FISH confirmed SRY and centromere of the Y chromosome on the questionable 13 chromosome and the karyotype to be 45,X,der(Y)t(Y;13)(q11.1;q12),-13.ish der(Y)(SRY+,DYZ3+,wcp13+).PCR-STS showed the deletion of regions AZFa,b and C,with a breakpoint located inYq11.1 below sY82.No deletion of the BRCA2 gene was observed.The patient was diagnosed with Sertoli cell-only syndrome by testicular biopsy and with angiolipomata by pathological examination of the nodule tissue.Conclusion: The patient's phenotype of complete masculinization could be attributed to presence of the SRY gene,and his azoospermia with small testis to the absence of a fragment from Yq11.1 to Yqter.However,the molecular mechanism of angiolipoma remains unknown.

OBJECTIVE: To study the pathological morphological changes for diagnosing the cause of death of extensive soft tissue injury or crush syndrome. METHODS: The tissues were stained by HE and IHC. RESULTS: (1) The Mb positive rate was 60%, 75%, 95% respectively. (2) Both the HSP70 positive rate of hearts and brains were 90%. CONCLUSION (1) The animal model of broad soft tissue injury was established. (2) Accumulated the pathological morphological data for diagnosing the cause of death of extensive soft tissue injury or crush syndrome.
Animals ; Cause of Death ; Crush Syndrome/pathology* ; Forensic Medicine ; HSP70 Heat-Shock Proteins/metabolism* ; Kidney/pathology* ; Myocardium/pathology* ; Myoglobin/metabolism* ; Rats ; Rats, Sprague-Dawley ; Soft Tissue Injuries/pathology*

OBJECTIVETo explore the retreatment of CHC patients with initial treatment failure and how to achieve SVR.

METHODS54 patients who had experienced treatment failure were enrolled and retreated with standard treatment of pegylated interferon and ribavirin or intensive treatment, respectively. Their SVR rates were statistically compared, to decide two therapies' application.

RESULTS54 patients had been retreated, and total SVR rate was up to 75.92%, with 88.46% in relapsed patients and 64.29% in non-responders. After retreatment with pegylated interferon and ribavirin, SVR rate was 95.45% in patients with prior interferon monotherapy, and 64.71% in patients with prior interferon and ribavirin, and 60% in patients with prior pegylated interferon alpha-2a monotherapy. SVR rate of relapsed patients was significantly higher than that of non-responders.

CONCLUSIONSIn CHC patients with treatment failure, SVR rate of retreatment with standard treatment or intensive treatment still can be up to 60%-90%. Retreatment with standard therapy can be applied to patients who had received interferon monotherapy or interferon plus ribavirin. Three types of patients who need intensive retreatment were as following: patients nonresponsive to interferon plus ribavirin or pegylated interferon alpha-2a monotherapy, and patients with treatment failure who had received prior standard treatment.

Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Female ; Hepacivirus ; drug effects ; genetics ; isolation & purification ; physiology ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; therapeutic use ; Retreatment ; Ribavirin ; therapeutic use ; Treatment Failure ; Viral Load ; drug effects ; Young Adult