Objective To investigate the comparison of clinical curative effect and inflammation factors between irbesartan hydrochlorothiazide and candesartan cilexetil in the treatment of patients with hypertension.Methods A total of 86 patients with hypertension in our hospital from June 2014 to October 2015 were collected and randomly divided into two groups, 43 cases in the control group were treated by candesartan cilexetil,43 cases in the experimental group were treated by irbesartan hydrochlorothiazide.The blood pressure, CRP, IL-6 and NO indexes were detected in the two groups and the clinical curative effect of the two groups was compared.Results The systolic pressure and diastolic pressure levels in the experimental group were significantly lower than the control group ( P <0.05 ); the serum CRP, IL -6 levels in experimental group were significantly lower than the control group, the NO level was significantly higher than the control group (P<0.05).Conclusion Irbesartan hydrochlorothiazide in the treatment of patients with hypertension has a better clinical curative effect, and higher security.

OBJECTIVES:The effect of water-soluble polymers polyvinylpyrrolidone(PVP) and polyethylene glycol(PEG)on the complexation of drugs with β-cyclodextrin were investigated.METHODS:the thermodynamic properties of drugs in aqueous β-CD solution and in aqueous solut.RESULTS:The maximum solubilization concentration of PVP or PEG appeared from 1.5g.L－1 to 3.0g.L－1.At this low concentration,PVP or PEG had insignificant effect on the viscosity of the aqueous β-CD solution.The solubilising effect ofβ-CD on chloramphenicol was improved by 15% when containing 2.5g.L－1 polyethylene glycol in the solution and on vitamininum B2by 98%when containing 2.5g.L－1 polyvinylpyrrolidone.In aqueous solutions water-soluble polymers were shown to increase the solubilising effect of β-CD on drugs.Additing of PEG or PVP to the aqueous complexation medium resulted in an increased negative enthalpy change,together with an increased negative entropy change.CONCLUSION：Water-soluble polymers were shown to increase the solubilising effect of β-CD on drugs.

Objective:To investigate the changes of gene expression profile in lipopolysaccharide (LPS)-stimulated primary Kupffer cells ( KCs ) . Methods: Rat KCs were isolated and purified by means of in situ perfusion and density gradient centrifugation. After being identified by ink phagocytosis and ED2 staining test,KCs were stimulated with LPS. Gene expression profile were studied using gene microarrays,and the most significant upregulated gene was verified using real-time PCR. Results:27 genes were upregulated including Ces1f, Slc17a3, Slc21a4, Hsd17b2, Sorbs2, Ccdc116, Mgam, Myo5b, Etl4, Fabp1, Kif4b, Fosl1, Cyp4a1, Penk, Tmem221,Rpl5,Nr2f1,Hoxb1,Gpr165,Fam90a13p,Kpna6,Irak1bp1,Kcnh1 and 4 unnamed genes and 4 downregulated including Oc90,Tagln,Arxes2 and Olr830 in LPS-stimulated KCs. Among the upregulated genes, Ces1f was the most significant upregulatory gene. Real-time PCR confirmed that the levels of Ces1f were 23. 88 times higher in LPS-stimulated than control cells. Conclusion:There is a significant difference between LPS-stimulated and normal control cells in gene expression profile by microarray analysis,and Ces1f is the most significantly upregulated gene.

Objective To investigate the impacts of adenovirus on the transfection efficiency and proliferative activity of primary Kupffer cells (KCs). Methods Rat liver KCs were separated and purified by density gradient centrifugation , and was then transfected with adenovirus carrying green fluorescence protein (GFP) gene at different multiplicity of infection (MOI). After 24 h, the transfection efficiency was evaluated by fluorescence microscope and flow cytometry. The proliferative activity of KCs was assessed by colorimetric method. Results The positive percentages of GFP staining cells were statistically different among different doses of adenovirus (MOI 0, 100, 300, 500, 700 and 900) under fluorescence microscopy or by flow cytometry (P <0.05 for all comparisons). The cell proliferative activity had significant differences among MOI 300, 500, 700 and 900(P < 0.05 for all comparisons), but had no differences among MOI 0, 100 and 300 (P > 0.05 for all comparisons) by CCK8 assay. Conclusions KCs can effectively be transfected by GFP adenovirus; and with an increase in virus MOI, the transfection efficiency rises gradually. A higher dose of adenovirus may have a negative effect on cell proliferative.

Objective To investigate the effects of interferon regulator factor 3 (IRF3) shRNA on the expression of TLR4 downstream signal molecules including IRF3-IFN-β, NF-κB/p38 MAPK-TNF-α/IL-1βand IL-10 in lipopolysaccharide (LPS)-stimulated Kupffer cells (KCs). Methods KCs were isolated from rats by in situ perfusion. The adenovirus strains carrying IRF3 shRNA were used for the transfection of purified KCs. The isolated KCs were randomly divided into four groups including adenovirus(-) LPS(-) treatment group, adenovirus(-) LPS(+) treatment group, adenovirus(+) LPS(-) treatment group and ad-enovirus(+) LPS(+) treatment group. The levels of cytokines in the supernatants of KC culture were detec-ted by enzyme-linked immunosorbent assay ( ELISA ) . Real-time PCR and Western blot assay were per-formed to analyze the expression of related cytokines at mRNA and protein levels, respectively. Results The expression of IRF3 at mRNA and protein levels in primary cultured KCs were induced by LPS. The cel-lular constitutive expression of IRF3 at mRNA level and the LPS-induced expression of IRF3 were signifi-cantly inhibited after transfection of KCs with adenovirus strains carrying IRF3 shRNA. However, the nucle-ar constitutive expression of IRF3 protein was not affected by IRF3 shRNA. The expression of IFN-βat mR-NA and protein levels in KCs were induced by LPS, but were suppressed by the interference with IRF3 shR-NA. No significant changes of the cellular constitutive expression of IFN-βat mRNA and protein levels were observed in IRF3 shRNA-treated KCs. Enhanced expression of proinflammatory cytokines including TNF-αand IL-1β at mRNA and protein levels were detected in LPS-stimulated KCs. Transfection of KCs with ade-novirus strains carrying IRF3 shRNA inhibited the LPS-induced secretion of TNF-α and IL-1β, but neither LPS-induced expression of TNF-α and IL-1β at mRNA level nor cellular constitutive expression of TNF-αand IL-1βat mRNA and protein levels were affected by IRF3 shRNA. The LPS-induced expression of IL-10 at mRNA and protein levels were enhanced in IRF3 shRNA-treated KCs. However, the cellular constitutive expression of IL-10 at mRNA and protein levels were not affected by the adenovirus. The levels of phosphor-NF-κB p65 subunit and phosphor-p38 MAPK protein in the nuclei of KCs were increased upon the stimula-tion with LPS. Treatment of KCs with IRF3 shRNA showed no significant effects on nuclear phosphor-NF-κB p65 subunit and phosphor-p38 MAPK. Conclusion Transfection of LPS-stimulated primary KCs with ade-novirus strains carrying IRF3 shRNA could effectively inhibit the expression of IRF3 and the transduction of downstream signals. IRF3 enhanced the secretion of TNF-αand IL-1β, but inhibited the expression of IL-10 in LPS-treated KCs. The LPS-induced activation of NF-κB and p38 MAPK in KCs were not affected by IRF3 signal.

Objective:To investigate the effects of IRF 3 shRNA adenovirus on dynamic changes of early cytokines in LPS-stimulated primary Kupffer cells ( KCs ).Methods: Rat KCs were isolated and purified by means of in situ perfusion.After being infected with adenovirus carrying IRF 3 shRNA for 48 h, KCs were stimulated with LPS.Cell culture supernatants were collected respectively at 0,2,4 and 6 h after LPS stimulation as well as cells at 6 h.Supernatant cytokine secretion levels were detected by enzyme-linked immunosorbent assay ( ELISA).Intracellular gene expressions were tested by RT-PCR and Westeron blot.Results:IRF3 mRNA and protein were induced by LPS ,but suppressed by IRF 3 shRNA adenovirus in LPS-stimulated or non-stimulated KCs.IFN-βsecretions rose in the very early stage ( at 2 h) ,reached the peak at 4 h,and began to reduce but still remained high levels at 6 h after LPS stimulation in KCs.Interference adenovirus pretreatment suppressed IFN-βsecretions ( especially the secretion peak ) at each time point after LPS stimulation.IFN-βsecretions reached normal levels at 6 h after the stimulation in adenovirus-pretreated cells;TNF-αse-cretions rapidly increased in the very early stage and reached the peak at 2 h,then began to decrease gradually ,but remained high levels at 6 h after LPS stimulation in KCs.Interference adenovirus pretreatment inhibited LPS-induced TNF-αsecretions, especially the secretion peak;IL-1βsecretions did not increase untill 4 h, but reached a higher level at 6 h after LPS stimulation.Interference adenovirus suppressed IL-1βsecretions in the early stage of LPS stimulation;IL-10 secretions began to rise in the very early stage ,and gradually increased over time after LPS stimulation in KCs.Pretreatment of adenovirus with IRF 3 shRNA promoted upregulations of IL-10 secretions at each time point of the early of LPS stimulation.Conclusion:IRF3 gene expression can be silenced by IRF 3 shRNA ad-enovirus.IRF3 can promote its downstream signaling molecule IFN-βand pro-inflammatory cytokines including TNF-αand IL-1β,and block anti-inflammation cytokine IL-10 secretions in LPS-stimulated primary KCs.Therefore,IRF3 may play a central role in immune inflammatory injury of liver tissues.

Objective Aδ and C fiber nociceptors can be excited when people received contact heat stimulation(CHS) and this excitation may be reflected by functional magnetic resonance imaging(fMRI).The aim of this study was to observe the different fMRI characteristics in CHS with distinct temperatures in healthy people and to explore the pain related functional network.Depending on the fMRI results,we can evaluate the values of CHS-fMRI during the research of brain functional connectivity.Methods Twenty-two healthy volunteers were recruited to this study.During the acquisition of fMRI,the right dorsal forearm received two different CHS in 41 and 51 degree respectively(41 ℃ CHS-fMRI group and 51 ℃ CHS-fMRI group),and two series of fMRI data were obtained for each subject.The brain activation was obtained by using one sample t test for the 41 ℃CHS-fMRI group and 51 ℃CHS-fMRI group separately.The difference between 41 ℃ CHS-fMRI group and 51 ℃ CHS-fMRI group was analyzed by paired-sample t-test.Result These activated brain areas in 41 ℃ CHS-fMRI group included bilateral superior temporal gyrus (STG),contralateral transverse temporal gyrus,contralateral amygdala,bilateral insula,bilateral inferior frontal gyrus (IFG),contralateral hippocampus,bilateral putamen,contralateral supramarginal,ipsilateral postcentral,ipsilateral inferior parietal lobule,ipsilateral middle temporal gyrus (MTG),bilateral cerebelum,bilateral middle cingulate cortex and bilateral posterior cingulate cortex.These activated brain areas in 51 ℃ CHS-fMRI group included bilateral STG,bilateral amygdala,contralateral hippocampus,bilateral thalamus,contralateral putamen,bilateral insula,bilateral IFG,bilateral cerebellum,contralateral postcentral,contralateral superior parietal lobule,bilateral MTG,bilateral precuneus,and contralateral cuneus.Compared with 41 ℃ CHS-fMRI group,the 51 ℃ CHS-fMRI group showed higher brain activation in bilateral cerebellum (right:4.455,left:3.891),ipsilateral precuneus (4.150),contralateral insula (3.530),contralateral IFG (3.530),and contralateral postcentral (3.530; t =2.83,P ＜ 0.01).Conclusions There are common brain activated areas and specific areas for each group,which suggested that existence of two central pathways activated by Aδ and C fiber which have different effects in perception of pain and have their brain network responsively.It may become one of the ideal pain stimulation methods with CHS-fMRI,which warrant worth further research.

Objective To investigate the alterations of regional spontaneous activity in patients with classical trigeminal neuralgia (CTN) during resting state.Methods Twenty-seven patients with CTN (CTN group) and 27 healthy subjects (control group) were recruited and underwent a rest-state functional MRI.The regional homogeneity (ReHo) analysis was used to compare the differences of regional synchronization of spontaneous brain activity.And correlation tests were performed between ReHo values in the abnormal brain areas and clinical metrics (visual analogue scale and disease duration) of the disease.Results Compared with control group (P＜0.05,Gaussian random field correction),ReHo increased in bilateral primary somatosensory cortex (S1) and primary motor cortex (M1),right supplementary motor area (SMA),inferotemporal cortex and cerebellum,left thalamus,limbic lobe,parahippocampa gyrus,middle and superior temporal gyrus in CTN group;ReHo decreased in bilateral insula,prefrontal cortex and orbitofrontal cortex,right frontal medial cortex and superior temporal gyrus,left anterior cingulate area,supramarginal gyrus and cerebellum in CTN group.ReHo values in right frontal medial cortex was negatively correlated with the course of disease (r=-0.45,P=0.03).The ReHo values of left primary sensorimotor cortex were positively correlated with the visual analogue scale scores (r=0.46,P=0.02).Conclusion CTN patients has abnormal functional homogeneity of spontaneous brain activity in regions involved in the pain processing,which can help understanding mechanism of CTN.

Objective: To learn the status and influencing factors of development among infants at ages of 0 to 36 months in Xiaoshan District, Hangzhou City, so as to provide the reference for promoting healthy development of infants. Methods: Infants at ages of 0-36 months who underwent physical examination in Child Health Clinic of Xiaoshan District Community Health Service Center were selected in 2022. General data of infants and their mothers were collected through questionnaires, and the development status of infants was screened by Age and Stages Questionnaire (third edition). Factors affecting the development status were identified using a multivariable logistic regression model. Results: A total of 2 519 infants were investigated, including 1 339 males (53.16%) and 1 180 females (46.84%). There were 608 infants with abnormal development of at least one functional area of communication (CM), gross motor (GM), fine motor (FM), problems solving (CG) and personal-social (PS). The abnormal rate was 24.14%, and the abnormal rates of the above functional areas were 9.77%, 6.59%, 7.98%, 6.39% and 9.33%, respectively. Multivariable logistic regression analysis showed that gender (male, OR=1.563, 95%CI: 1.191-2.052), mother's childbearing age (≥35 years, OR=1.411, 95%CI: 1.001-1.988), mother's educational level (lower than junior college, OR=1.460, 95%CI: 1.116-1.912) were factors affecting abnormal development of CM; preterm birth (OR=2.323, 95%CI: 1.315-4.103) was factors affecting abnormal development of GM; gender (male, OR=1.654, 95%CI: 1.225-2.232) was factors affecting abnormal development of FM; gender (male, OR=1.511, 95%CI: 1.086-2.102) and mode of delivery (cesarean section, OR=1.460, 95%CI: 1.060-2.010) were factors affecting abnormal development of CG; gender (male, OR=1.340, 95%CI: 1.019-1.763) and birth weight (low birth weight, OR=1.985, 95%CI: 1.149-3.432) were factors affecting abnormal development of PS. Conclusions The rate of abnormal development among infants at ages of 0 to 36 months in Xiaoshan District is 24.14%. Gender, preterm birth, mode of delivery, birth weight, mother's childbearing age and mother's educational level could affect the development status of infants.

OBJECTIVE: To explore the effect of the oxidored nitro domain containing protein 1 (NOR1) gene knockdown on the biological behavior of HeLa cells in cervical carcinoma. METHODS: The recombinant plasmids pSUPER-shNOR1-1, pSUPER-shNOR1-2 and pSUPERscramble, which targeted to NOR1 gene, were constructed by pSUPER.neo+GFP vector, transfected into HeLa cells respectively using Lipofectamine 2000 reagent, and followed by G418 selection. The expression level of NOR1 mRNA and protein were determined by RT-PCR and Western blotting, respectively. Methyl thiazolyl tetrazolium (MTT) assay was performed to determine the growth curve of cell viability. The stable transfectants were treated with H₂O₂ and cell apoptosis was determined by Hoechst 33258 staining and terminal deoxynucleotidyl transferasemediated dUTP nick end labeling (TUNEL) assay. The expression levels of Bcl-2, cleaved caspase 9 and poly ADP-ribose polymerase (PARP) were measured by Western blot. RESULTS: NOR1- knockdown HeLa cells were successfully constructed by transfection of pSUPER-shNOR1-1 or pSUPER-shNOR1-2 plasmids into HeLa cells. MTT assay showed that the silence of endogenous NOR1 in HeLa cells could lead to the increase in cell viability and proliferation, and the inhibition of H₂O₂-induced apoptosis compared with the negative control. Western blot showed that the expression level of active caspase 9 and cleaved PARP was inhibited in NOR1-knockdown cells when they were treated with H₂O₂ while the expression level of Bcl-2 protein increased. CONCLUSION Silence of endogenous NOR1 facilitates the cell viability and growth of HeLa cells, and attenuates HeLa cells apoptosis induced by H₂O₂, which might be mediated by up-regulation of Bcl-2 level and down-regulation of the cleaved caspase 9 cascade.
Apoptosis ; Caspase 9 ; metabolism ; Cell Survival ; Down-Regulation ; Gene Knockdown Techniques ; Genetic Vectors ; HeLa Cells ; Humans ; Hydrogen Peroxide ; Membrane Transport Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; Transfection ; Up-Regulation