Objective One hundred and sixteen cases of pediatric patients with severe heart fail-ure,pneumonia combined application of phentolamine dopamine synthesis with conventional medical treat-ment were compared the clinical efficacy,analyze and judge the feasibility of both combination therapy clini-cal significance and its value.Methods Collected from July 2013 to April 2015 period to the author hospi-tal and diagnosed with pneumonia in children with heart failure in 116 patients.Which were randomly divid-ed into two groups,group A +phentolamine medical treatment with dopamine and B group routine clinical medicine comprehensive treatment.Clinical observation by a number of factors to analyze comparative A,B two groups after the treatment efficacy.Results A significant effect of the treatment group and the number of effective treatments were more than group B and A group of overall response rate (91.38%)than in group B (60.34%);A group invalid proportion treated patients (8.62%)was significantly less in group B (39.66%);A group of children mortality (5.17;Change a group on heart rate and blood pressure had a better than group B;and the difference was greater (P <0.05).A,B two members appeared after treat-ment of mild nasal congestion and mild bloating and other adverse reactions,for Group A after two cases of children with nasal 1% solution of furosemide Ma nasal symptoms disappear;for two cases of group B chil-dren with mild bloating conventional medical treatment after symptoms have been effectively controlled.A, B two groups in terms of adverse events was not significantly (P >0.05)Conclusions A group of overall efficiency significantly higher in group B and body recover faster in children,low mortality,children with severe pneumonia prompted phentolamine and dopamine treatment with heart failure better.

Objective To investigate the expression and significance of Rho-associated protein kinaseⅡ(Rock Ⅱ)in preeclamptic placenta and umbilical artery.Methods Semiquanfitative RT-PCR and Western blot were used to investigate the expression of RockⅡmRNA and RockⅡprotein in placenta and umbilical artery from 35 women with moderate preeclampsia(MPE group)、38 women with severepreeclampsia(SPE group)and 45 normal third trimester pregnant women(control group),the S/D value of umbilical artery was examined by ultrasound.Results (1)The expression of Rock Ⅱ mRNA of Dlacenta in MPE group(0.82±0.14)and SPE group(0.93±0.13)were signifieantly higher than that in control group (0.70 ±0.12,P＜0.01).The expression of Rock Ⅱ protein of placenta in MPE group(0.79±0.15)and SPE group(0.92±0.12)were significantly higher compared with control group(0.68±0.11,P<0.01).The expression of Rock Ⅱ mRNA and protein of placenta in SPE group were higher compared with MPE group(P＜0.01).(2)The expression of Rock Ⅱ mRNA of umbilical artery in MPE group(0.69±0.13)and SPE group(0.55±0.12)were significantly lower than that in control gmup(0.76±0.10,P＜0.01).The expression of RockⅡ protein of umbilical artery in MPE group(0.68±0.10)and SPE group(0.51±0.12)were lower compared with control group(0.75±0.13,P＜0.01).The expression of RockⅡ mRNA and protein of umbilical artery in SPE group were significantly lower compared with MPE group(P＜0.01).(3)There were no correlations between the expression of RockⅡ mRNA and protein in placenta and umbilical artery and the S/D value and birth weight(P＞0.05).Conclusion The upregulated expression of Hock Ⅱ in placentas and downregulated expression in umbilical artery may be a compensation in preeclampsia.

Aim To construct eukarytic vector for rat endostatin(endos) cDNA and observe its expression in C6 cell.Methods cDNA encoding rat endostatin was amplified from newborn brain tissue with RT-PCR and inserted into the eukarytic vector pBudCE 4.1.Recombinant was identified with KpnI,XhoI double digestion,PCR and nucleotide sequencing of the target gene.After successful reconstruction of the genes of endostatin,the recombinants was transfected into C6 cells with lipofectintechniques.The positive clones were screened out through zeocin resistance test.The endostatin in supernate of the positive clones was identified with Western-blot and MTT method.With immunocytochemistry,the endostatin in the positive clones was located.The quantities of VEGF in supernate of the positive clones were quantified with ELISA assay.Results The size of the amplified endostatin gene fragment was in accord with that we expected.And the gene sequence inserted into the eukarytic vector pBudCE 4.1 was consistent with the known sequence.Endostatin was secreted from the positive clone.Down-requlation of vascular endothelial growth factor(VEGF) was found in the positive clones.Conclusion The recombinant of rat endostatin gene clone had been established and inserted into the eukarytic vector pBudCE 4.1 successfully and endostatin was expressed in C6 cells.This provides a basis for further studies of endostatin effects in vivo,and creates the conditions for final clinical trial

Aim To investigate the influence of endostatin on pathological morphology of C_6 glioma and its molecular pharmacologic mechanism.Methods The C_6 cells,the C_6 clone stably transfected with endostatin cDNA(endo-C_6),which the endostatin with biological activities can be secreted from,and the C_6 clone stably transfected with empty vector pBudCE4.1 cDNA(pBud-C_6) were injected subcutaneous in nude mice to establish different tumoral model respectively.The morphologies of these tumoral tissues were observed and compared to each other under light and electron microscope.The expression of VEGF in tumor tissue was determined by ELISA.Results The expression of VEGF in endo-C_6 glioma(endo-C_6G) tissues was lower than that in C_6 glioma(C_6G) and pBud-C_6 glioma(pBud-C_6G).Moreover,endo-C_6G tissue was characteristic of a mimetic envelope,no intratumor bleeding and cystis degeneration,apoptosis of tumor cells,and edema in and around tumor.Rarefied vessels were found in tumor,and no vessel like structure formed by tumor cells was observed.The large irregular necrosis focus was showed in tumor,but mild vascular reaction around necrosis focus and peritumor,rare surrounding invasion.The basal lamina was discontinued.The basemembrane(BM) was loose.Few vesicular vacuolar organelle(VVO) structures were observed in plasma of endothelial cells.In C_6G and pBud-C_6G,tumor lesions demonstrated significant vascular reaction,intratumor bleeding,necrosis,edema in and around tumor,and surrounding infiltration.Vessel like structure formed by tumor cells was also observed.When examined with electron microscope,plenty of VVO structures were observed in plasma of endothelial cells,the worse the edema,the more the VVO were,which coincided with the expression of VEGF.Mostly,loose basal lamina surrounded by small amounts of collagen fibers was multilayer and integrated and continuous.No correlation between gene transfection and fenestra formation or cleft of capillary endothelial cell was observed,no apoptosis of endothelial cells were found.Conclusion In glioma,the apoptosis of endothelial cells tissue was not induced directly by endostatin,but the angiogenesis and vascular reaction can be inhibit by endostatin by down-regulation of the expression of VEGF in C_6 glioma cells.

ObjectiveTo determine the subcellular localization of exogenous human papillomavirus type 16 E6 protein(HPV16 E6) and hDaxx in HeLa cells and their effects on tumor necrosis factor (TNF)-α-induced apoptosis.MethodsHeLa cells were transfected with plasmids pDsRed-monomer-C1/HPV16 E6,pEGFP-CI/hDaxx,pEGFP-C1 and pDsRed-monomer-C1 respectively.Subsequently,Western blot was carried out to quantify the expression of fusion proteins DsRed-HPV16E6 and EGFP-hDaxx in transfected cells,and laser scanning confocal microscopy to observe the subcellular distribution of HPV16 E6 protein and hDaxx.Some HeLa cells were divided into 5 groups:untransfected (control group),untransfected and treated with TNF-α(TNF-ot group),transfected with pcDNA3.1 (-) and treated with TNF-α(empty vector group),transfected with pcDNA3.1 (-)/HPV16 E6 and treated with TNF-α (HPV16 E6 group),cotransfected with pcDNA3.1(-)/HPV16 E6 and pcDNA3.1 (-)/hDaxx and treated with TNF-α (cotransfected group).After additional culture,the cells were collected and subjected to flow cytometry(FCM) to evaluate the apoptosis of cells as well as spectrophotometry to determine the relative activity of Caspase-8 and Caspase-3.ResultsWestern blot showed that both DsRed-HPV16 E6 and EGFP-hDaxx were expressed in HeLa cells.In Hela cells transfected with pDsRedmonomer-C1/HPV16 E6 or pEGFP-C1/hDaxx alone,the red fluorescence of HPV16 E6 was observed in the nucleus and cytoplasm,while the green fluorescence of hDaxx only in the nucleus; in those cotransfected with pDsRed-monomer-C1/HPVl6 E6,HPV16 E6 and hDaxx proteins were regionally aggregated near the nuclear membrane in nuclei,and hDaxx was partly translocated from the nucleus to the cytoplasm.The apoptosis rate and relative activity of Caspase-8 and Caspase-3 were statistically lower in HPV16 E6 group than in the empty vector group and cotransfected group(21.4％ ± 1.1％ vs.27.0％ ± 0.9％ and 32.5％ ± 2.1％,0.057 ± 0.003 vs.0.092 ±0.012 and 0.109 ± 0.013,0.054 ± 0.006 vs.0.093 ± 0.005 and 0.110 ± 0.004,all p＜ 0.01).Conclusions HPV16 E6 protein induces the partial translocation of hDaxx from the nucleus into the cytoplasm and colocalizes with hDaxx in the cells.The apoptosis of HeLa cells induced by TNF-α can be suppressed by HPV16 E6 protein,while the overexpression of hDaxx can attenuate the suppressing effect of HPV16 E6 protein on apoptosis in Hela cells.

Objective To investigate the clinical effect of rhG-CSF combined transplantation of autologous bone marrow mesenchymal stem cells on cerebral infarction. Methods 42 acute cerebral infarction patients were randomly divided into tow groups: Injection rhGCSF combined autologous bone marrow mesenchymal stem cells group(treatment group, n=20)and conventionality therapy group(control group, n=22).The efficacy was assessed by National Institutes of Health Stroke Scale(NIHSS), Barthel Index(BI)and diffusion-weighted imaging(DWI),perfusion-weighted imaging(PWI)at baseline and the 3rd month after treatment. Result The scores of NHISS and BI treatment group were 4.8±2.0 and 78.5±7.2 respectively. while were 7.2±2.4 and 56.1±6.3 in control group at the 3rd month after treatment. The difference were significant(P＜0.01).PWI in treatment group Was higher than that in control group. There was no significant side-effect in treatment group. Conclusion rHG-CSF combined autologous bone marrow mesenchymal stem cells is a safe, efficient treatment for acute cerebral infarction patients, which is a better way than conventionality therapy.

Objective To find out the distribution of drinking-tea-borne fluorosis in the six ethnics in Qinghai Province, and to provide basic data for prevention and control of the disease. Methods In 2010, according to the requirement of “The National Surveillance Program of Drinking-Tea-borne Fluorosis”, six ethnics accounted for 99.59% of total population in Qinghai Province were investigated in 28 counties having brick-tea drinking habit. Three townships and a town in each county, two administrative villages(residents’ committee) in each township and town were chosen and 50 adults in each administrative village and residents ’ committee were selected to check skeletal fluorosis, dental fluorosis, urine fluoride and daily drinking amount of tea water. Five to six samples of drinking tea water were determined. Dental fluorosis was examined by Deans method; the fluoride content of brick-tea and urine were determined by fluoride ion selective electrode; the skeletal fluorosis was diagnosed based on “Endemic Osteofluorosis Clinical Indexing Diagnosis Standard”( WS 192-2008 ) . Results A total of 10 335 adults were surveyed, the number of Tibetan, Han, Hui, Mongolian, Tu and Salar ethnics were 4 972, 3 063, 1 196, 634, 235 and 235, respectively. The daily drinking amounts of tea water in Mongolian, Tibetan, Hui, Tu, Han and Salar ethnics were 2.53, 2.19, 1.74, 1.63, 1.22 and 1.07 L, respectively. Daily fluoride intakes in Tibetan, Mongolian, Tu, Hui, Han and Salar ethnics were 3.99, 2.78,2.27, 2.16, 1.78 and 1.28 mg, respectively. The medians of urinary fluoride concentration of the Tibetan, Tu, Hui, Han, Mongolian and Salar ethnics were 1.46, 1.19, 1.12, 0.98, 0.93 and 0.81 mg/L, respectively. The prevalence rates of dental fluorosis of the Hui, Han, Tibetan, Tu, Mongolian and Salar ethnics were 34.53%(413/1 196), 27.07%(829/3 063), 21.60%(1 074/4 972), 20.00%(47/235), 17.98%(114/634) and 6.38%(15/235), respectively. The incidence rates of clinical skeletal fluorosis of the Tibetan, Mongolian, Han, Hui, Tu and Salar ethnics were 13.42%(667/4 972), 11.04%(70/634), 9.31%(285/3 063), 7.61%(91/1 196), 5.53%(13/235) and 4.26%(10/235), respectively. Conclusions The distribution and prevalent status of drinking-tea-borne fluorosis in the six ethnics of Qinghai Province are different. Tibetan and Mongolian ethnics are the key population concerning the prevention and control of the disease.

Objective:To investigate whether pORF5 plasmid protein of Chlamydia trachomatis(Ct) induces 1L-1βand 1L-18 production in THP-1 cells,and its potential molecular mechanism.Methods:pORF5 plasmid protein was used to stimulate THP-1 cells at different concentrations(0,3,6,12,24,36 μg/ml),then the inflammatory cytokines IL-18 and IL-1βwere detected by ELISA at the time of 0,8,16,24,36 h;The mRNA expression of NALP3 inflammasome were detected by Realtime-PCR,and Caspase-1 activity was determined by Western blot analysis.THP-1 cells were transfected with siRNA targeting NALP3 and ASC gene for 24 h or pretreated with Caspase-1 inhibitor(Z-YVAD-FMK) for 30 min,and subsequently stimulated with pORF5(24 μg/ml) for 24 h,then secretion of IL-1βand IL-18 were analyzed by ELISA.Results: The pORF5 plasmid protein induced THP-1 cells to secrete IL-1βand IL-18 by dose-and time-dependent manners,production of IL-1βand IL-18 reached their peaks(491 pg/ml and 186 pg/ml) at concentration of 24 μg/ml,and the peak amount of IL-1βand IL-18 occurred at 24 h and 16 h post-stimulation respectively.pORF5 plasmid protein in-creased mRNA expression of NALP3 inflammasome and activated Caspase-1 in THP-1 cells.NALP3 siRNA,ASC siRNA and Z-YVAD-FMK reduced pORF5-induced IL-1βand IL-18 production when compared with control groups(P<0.05).Conclusion:pORF5 plasmid protein could induce THP-1 cells to produce IL-1βand IL-18 through NALP3 inflammasome activation,which may play an important role in the pathogenesis in Ct infection.

Objective To observe the illness change trend of drinking water type endemic arsenic poisoning in Qinghai Province, comprehensively evaluate the effect of prevention and control measures, in order to provide a scientific basis for timely adjustment of the prevention and control measures. Methods From 2010 to 2013, according to the Drinking Water Type Endemic Arsenic Poisoning Monitoring Programs, 3 villages in 2 counties within Qinghai Province were randomly selected as monitoring sites. The arsenic content in drinking water of residents was measured, water improvement projects in all monitoring villages were investigated; at the same time an investigation of arsenic disease in resident population was conducted, and urinary arsenic content was monitored. Arsenic in drinking water and urine was determined by hydride generation atomic fluorescence spectrometry, and arsenic poisoning was diagnosed using Endemic Arsenic Poisoning Diagnostic Criteria (WS/T 211-2001). Results Of the three water improvement projects, two were water arsenic exceeded the standard, and one was intermittently operated. From 2010 to 2013, the arsenic poisoning detec tion rate in Baoning Village was 27.30% (193/707), 31.90%(245/768), 29.35%(221/753) and 28.22%(219/776); in Kecai Village was 32.62%(107/328), 34.83%(124/356), 31.26%(131/419) and 29.35%(118/402);and in Manimotai Village was 56.58%(43/76), 52.81%(47/89), 45.10%(46/102) and 34.69% (34/98), there was no significant difference statistically of the arsenic poisoning detection rates in the three monitored villages in the 4 years (χ2 =3.09, 0.04, 0.92, all P>0.05). From 2010 to 2013, women arsenic poisoning detection rate was 36.45%(203/557), 40.59%(246/606), 36.12%(225/623) and 34.77%(218/627), respectively;men was 25.27%(140/554), 28.01%(170/607), 26.57%(173/651) and 23.57%(153/649), respectively;women arsenic poisoning detection rates were higher than those of men (χ2 = 16.25, 21.32, 13.49, 19.38, all P < 0.05). Arsenic poisoning detection rate of people younger than 60 years old had a tendency to increase with age. In 2012 and 2013, 105 and 93 urine samples were tested, respectively; urinary arsenic geometric mean was 0.113 and 0.149 mg/L. Conclusions Water improvement projects and water quality are not optimistic, and prevalence of arsenic poisoning is still at a higher level. A sound long-term monitoring program should be established as soon as possible, the management and maintenance of water improvement projects should be strengthened, and the monitoring and prevention work should not be neglected.