Objective To analyze the risk factors for hypertension in patients with chronic kidney disease stage 5(CKD5) . Methods The basic information of 390 CKD5 patients complicated with hypertension was collected for univariate analysis ,in‐cluding gender ,age ,primary disease ,dialysis method ,body mass index(BMI) ,complications(hyperlipidemia ,high uric acid ,car‐diac insufficiency) ,level of education ,parathyroid hormone(PTH)level.Univariate variables that showed statistical significance were then subjected to the multivariate analysis(Logistic regression)to identify the risk factors for hypertension in CKD5 pa‐tients.The defined daily dose(DDD)that satisfied the criteria interms of different stages was evaluated.Results Overall hyper‐tension control rate was 22.8%.Univariate analysis showed that the following variables were significantly associated with hy‐pertension in CKD5 patients :>40 years old ,male ,diabetic nephropathy ,hypertensive nephropathy ,hemodialysis ,hyperlipemia , high uric acid level ,and high PTH level(P<0.05).Logistic multivariate analysis showed that diabetic nephropathy ,hyperlipi‐demia ,high PT H level were the independent risk factors for hypertension in patients with CKD5.In hypertension segmented standard ,there was no difference in the DDD between stage 0 and 1(P>0.05) ,and DDD at stage 2 and 3 was increased signifi‐cantly when compared with that at 0 and 1 standard(P<0.05).Conclusion Overall hypertension control rate is very low in pa‐tients with CKD5.Diabetics ,hyperlipidemia ,high PTH level are independent risk factors for hypertension in patients with CKD5.

A software system of the pulse oximeter is presented in this paper. Based on Franklin C, this software system includes three main parts, one part is to automatically regulates the base line of signal, the second part is a controlled integral module,and the third part is a digital signal processing module. As the result, the pulse oximeter is satisfactory to clinical monitoring.

ObjectiveTostudytheroleof hotshockprotein (HSP)47in tubulointerstitial fibrosis induced by transforming growth factor β1(TGF-β1),and to explore its possible mechanism.Methods Human proximal tubular epithelial cells(HK-2) were divided into threegroups:control,TGF-β1andHSP47siRNA. Theexpressionsof HSP47, collagenⅣ,fibronectin(FN),plasminogen activator inhibitor 1(PAI-1) mRNA and HSP47,collagen Ⅳ,FN protein were detected by RT-PCR and Western blotting respectively.PAl-1 protein was detected by ELISA. ResultsHK-2expressedHSP47innormalmedium. ThemRNAandprotein expressions of HSP47 up-regulated in concentration- and time-dependent manner in HK-2 cells induced with increasing concentrations of TGF-β1(0,2.5,5,10 μg/L) and with prolong times (12,24,48 h),and peaked at 10 μg/L TGF-β1 for 48 h.Similar phenomena was observed in the mRNA andproteinexpressionsof collagenⅣ, FN, PAI-1inHK-2 cellsinducedbyincreasing concentrations of TGF-β1 (0,2.5,5,10 μg/L) at different time points (12,24,48 h),and peaked at 10 μg/L TGF-β1 for 48 h.HSP47 siRNA could significantly reduce the up-regulation of mRNA and protein expressions of HSP47,collagen Ⅳ,FN,PAI-1 in HK-2 cells induced by TGF-β1.Conclusion HSP47 can promote renal tubulointerstitial fibrosis maybe through the regulation of the expressions of collagen Ⅳ,FN,PAI-1.

Objective To evaluate the protective effects of mycophenolate mofetil (MMF) on the kidneys of diabetic rats and elucidate the associated mechanisms. Methods Wistar rats were divided into three groups: normal control rats, diabetic rats, and diabetic rats received and blood glucose were measured, and kidney pathology was observed. Inmmunohistochemistry and RT-PCR were used to analyze the expression of matrix metaUoproteinase-9 (MMP-9) and transforming growth factor 151 (TGF-β1). Results As compared with normal control rats, the 24 h urinary albumin excretion [(26.80±0.82) mg vs (6.64±1.42) mg], blood glucose[(22.18±3.36)mmol/L vs (6.40±0.87) mmol/L], Ccr [(0.220±0.380) ml/min vs (0.098±0.015) ml/min] of the diabetic rats were rised remarkbably. The 24 h urinary albumin excretion [(16.17±1.15) mg] and Ccr [(0.207±0.377) ml/min] of the diabetic rats received MMF were lower as compared to diabetic rats (P<0.05). MMP-9 in renal tissue of normal control rats was mainly expressed in glomerular mesangial ceils and renal tubular epithelial cells. Such MMP-9 expression was weak in diabetic rats and improved in the diabetic rats received MMF. There were significant differences among 3 groups. The expression of TGF-β1 was on the contrary. Conclusion Mycophenolate mofetil decreases 24 h urinary albumin, Cer and glomerular volume, which may be associated with the increase of MMP-9, the decrease of TGF-β1 expression and extracellular matrix deposition in renal tlssue.

Objective To observe the release of inflammation-related factors after angiotensin Ⅱ (Ang Ⅱ ) stimulation in rat tubular epithelial cells (NRK-52E), to analyze whether these effects were mediated by TLR4-MyD88 pathway, and to reveal the novel mechanism of injury by Ang Ⅱ on NRK-52E cells. Methods After synchronization, cells incubated with AngⅡ (10-7 mmol/L) were used as the stimulation group, cells without stimulation were as normal control. To determine the role of TLR4 and the adaptor MyD88, equal number of NRK-52E cells was added with 10-5 mmol/L candesartan or 20 mg/L TLR4 blocking peptide for 1 h and then incubated with Ang Ⅱ (10-7 mmol/L) respectively. RT-PCR was used to analyze TLR4 mRNA and MyD88 mRNA expression. Immunofluorescence and confocal microscopy were used to observe TLR4 protein expression. ELISA was used to detect the concentration of tumor necrosis factor-alpha (TNF-α) and heat shock protein 47(HSP47) in cell supernatant respectively. Results TLR4 and MyD88 were highly expressed in Ang Ⅱ-induced NRK-52E cells (P＜0.01), and the TNF-α and HSP47 levels were also increased markedly compared with control group (P＜0.01). In NRK-52E cells that were pre-incubated with candesartan, TLR4 and MyD88 expression were obviously inhibited,subsequently, HSP47 and TNF-α production decreased remarkably compared with Ang Ⅱ group (P＜0.01). TLR4 blocking peptide had the similar effect in a dose-dependent manner, in which its effect was dependent on inhibiting TLR4-MyD88 expression. Conclusion The mechanism of Ang Ⅱ -induced injury effect on NRK-52E cells is related to the increase of TLR4-MyD88 activity,which is followed by the enhance of TNF-α and HSP47 expression. This process is inhibited by candesartan via modulation of innate immune pathway.

Objective To observe the expression of toll like receptor 4(TLR4) Signaling and the release of inflammation factors in rat tubular epithelial cell(NRK-52E) under high glucose condition after TLR4-siRNA transfection.Methods Three TLR4-siRNA sequences were designed and synthesized.The transfection efficiency was observed by fluorescence microscope after transfection,and the expression of TLR4 mRNA was detected by real time PCR.The most effective siRNA was selected to be used for forward experiments.After transfection for 24 h,cells were stimulated with 25 mmol/L glucose and/or 10-7 mmol/L Angiotension Ⅱ (Ang Ⅱ) for 12 h,24 h; cells without stimulation were as normal control.Real-time PCR was used to analyze TLR4 and myeloid differentiation factor 88 (MyD88) mRNA expression; Western blot was used to observe TLR4/MyD88 and NF-κB protein expression.ELISA assay was used to detect the concentration of monocyte chemoattractant protein-1 (MCP-1),interleukin-6(IL-6) in cell supernatant after cells were stimulated for 24 h.Results TLR4/ MyD88 mRNA and TLR4/MyD88/NF-κB protein were highly expressed under high glucose or Ang Ⅱ co -incubated NRK-52E(P ＜ 0.01),the MCP-1 and IL-6 levels were also increased markedly compared with normal control group (P ＜ 0.01).TLR4/MyD88 mRNA and TLR4/MyD88/NF-kB protein expressions were obviously inhibited in cells that were transfected with TLR4-siRNA compared with high glucose group(P ＜ 0.01),MCP-1 and IL-6 production decreased remarkably compared with high glucose or Ang Ⅱ co-stimulated group(P ＜ 0.01).Conclusions High glucose can lead to the activation of TLR4/ MyD88/NF-kB signaling and the secretion of inflammation factors in NRK-52E,Ang Ⅱ further augments these effects.The effect can be blocked efficiently by specific siRNA gene silence.TLR4 signaling plays a pivotal role in the innate-immune inflammatory reaction in NRK-52E.

Objective To observe the regulation of Toll-like receptor 4 (TLR4) signal and the release of inflammation factors after angiotensin Ⅱ (Ang Ⅱ) stimulation in rat mesangial cells under high glucose condition,revealing the innate immune-related mechanism of injury by Ang Ⅱ on mesangial cells under high glucose.Methods After synchronization,cells incubated with Ang Ⅱ (10-7 mmo/L) and/or high glucose (25 mmol/L) were used as the stimulation group,cells without stimulation were as normal control (5.6 mmol/L glucose).To determine the role of TLR4 and the adaptor myeloid differentiation factor 88 (MyD88),equal number of HBZY-1 cells were added with 10-5 mmol/L irbesartan and/or TLR4 blocker (10 mg/L) for 1 h and then incubated with Ang Ⅱ (10-7 mmo/L) and/or high glucose (25 mmol/L) for 12 h or 24 h respectively.Real-time PCR was used to analyze TLR4 mRNA and MyD88 mRNA expression after 12 h.Immunofluorescence was used to observe TLR4 protein expression after 24 h; Western blotting was used to observe TLR4,MyD88 and nuclear factor κB (NF-κB) protein; ELISA was used to detect the concentration of MCP-1,IL-6 in cell supernatant respectively.Results Compared with normal control group,TLR4 mRNA and MyD88 mRNA were highly expressed in high glucose or Ang Ⅱ-induced HBZY-1 cells (P ＜ 0.01),TLR4,MyD88 and NF-κB protein as well as MCP-1,IL-6 were also up-regulated significantly (P ＜ 0.01).Compared with high glucose or Ang Ⅱ group,MyD88 and NF-κB protein as well as MCP-1,IL-6 were further up-regulated markedly in Ang Ⅱ and high glucose costimulated group (P ＜ 0.01).In HBZY-1 cells that were preincubated with irbesartan and/or TLR4 blocker,TLR4 and MyD88 protein expression were obviously inhibited,IL-6 and MCP-1 production were also decreased remarkably compared with high glucose and/or Ang Ⅱ group (P ＜ 0.01).Conclusions High glucose and Ang Ⅱ stimulate the release of proinflammatory factors in rat glomerular mesangial cells via TLR4-MyD88 pathway.This process is inhibited by irbesartan or TLR4 blocker via modulation of the signal.Ang Ⅱ has the positive-regulation potential on the release of inflammation factors via TLR4 signal in rat mesangial cells under high glucose condition.

Objective To investigate the effete of chitosan on rabbit carotid artery internal jugular vein fistula intimal hyperplasia and its regulation on TLR4/NF-κB signaling.Methods A total of 28 New Zealand white rabbits were randomly divided into the control group(n=4),the model group(n=12) and the chitosan group(n=12).Model group and chitosan group rabbits were established respectively carotid artery internal jugular vein fistula models.After AVF surgery,chitosan was smeared on venous blood vessels and anastomosis.After 4,6 and 8 weeks,the rabbits were separately sacrificed and the AVF venous vascular tissues were taken.The pathological changes of AVF venous vascular tissue in each group were observed.The changes of α-SMA were detected by immunohistochemistry method.The mRNA expressions of PCNA and TLR4 in the tissues were measured by Real-time PCR.At the same time,the protein expressions of PCNA,TLR4,MyD88 and NF-κB were detected by Western blotting.The experimental data were processed by two-factor analysis of variance in statistics.Results (1) After 4 weeks,vascular intimal was thicked in mdel group.In intimal hyperplasia,α-SMA was staining,and then proliferation of vascular smooth muscle cell was significant.As time increasing,more intimal hyperplasia shown obviously,the expression of α-SMA significantly increased.Compared with model group,chitosan group significantly reduced the degree of intimal hyperplasia,the level of α-SMA was significantly decreased,vascular smooth muscle cell proliferation was also extraordinarily decreased.(2) Compared with control group,the expression levels of PCNA,TLR4,MyD88 and NF-κB increased with time.The indices of Chitosan group were markedly higher than control group,but significantly lower than model groups.Conclusion Chitosan can inhibit the proliferation of rabbit VSMCs.The mechanism may be concerned in down regulating TLR4-mediated signaling pathway,reducing the possibility of intimal hyperplasia of rabbit AVF venous blood vessels.

Objective To explore the effect of chitosan on vascular smooth muscle cells inhibited proliferation from rabbit arteriovenous fistula and its mechanisms.Methods Established rabbit fistula model on carotid arteryinternal jugular vein.After 1 month cultured VSMCs with primary culture by tissue-pieces inoculation.Cultured VSMCs were divided into three groups:①normal control group.②FBS-treated group:cell were treated with 5％,10％,20％ for 48 h,respectively; established the model of rabbit VSMCs proliferation.③chitosan-treated group:VSMCs cultured with 20％ FBS were exposed to different doses of chitosan(10,100,500,1000,2000μg/ml) for 48 h.And VSMCs were treated for different time (0,12,24,48 h) with Chitosan 1000 μg/ml.Expression levels of PCNA and TLR4/ NF-κB were detected by Western blotting.RT-PCR were applied to measure the mRNA expression of PCNA and TLR4.The protein levels of TLR4 and NF-κB were detected by immunofluorescence.Results Compared with low concentration serum group,FBS-treated VSMCs exhibited a increase in mRNA and protein expression of PCNA and TLR4.FBS-induced protein expression of PCNA and TLR4/NF-κB were reduced by chitosan.Also mRNA expression of PCNA and TLR4 were reduced.They were dependent on concentration and time.In rabbit VSMCs TLR4 was mainly expressed in the cytoplasm and NF-κB expressed mainly in the nucleus.Compared with normal control group,TLR4 and NF-κB protein expression were significantly decreased by chitosan.Conclusion High concentration serum induced VSMCs proliferation.Chitosan can inhibit the proliferation of rabbit VSMCs.It is speculated that the mechanism may be related to the expression of TLR4 receptor activation,reducing expression of downstream factor MyD88 and NF-κB.It is suggest that chitosan can become potential new drugs of arteriovenous fistula prevention of intimal hyperplasia.

Objective To investigate the effect of suppressor of cytokine signaling 3 (SOCS3)on the proliferation of human mesangial cells stimulated by aggregated IgA1 (aIgA1) from patients with IgA nephropathy(IgAN),and explore its possible mechanism.Methods Serum monomeric IgA1 was isolated with jacalin affinity and Sephacryl S-200 HR chromatography from IgAN patients,and then heated to aggregated form (aIgA1).Human glomerular mesangial cells(HMC) were transfected with AdvSOCS3-IRES2-EGFP for 48 hours,and incubated with aIgA1 for 12-48 h.The cells were divided into blank control group,IgA1 group,IgA1 +Adv-EGFP group and IgA 1 +Adv-SOCS3-IRES2-EGFP group.The mesangial cell proliferation was observed through MTT,and the levels of SOCS3,TLR4,TGF-β1 protein and mRNA were detected through Western blotting and real-time PCR.Results HMC proliferation was promoted significantly after IgA1 stimulated at 24 h.Compared with control group,the protein and mRNA expression of SOCS3,TLR4,TGF-β1 were significantly increased in IgA1 group (P ＜ 0.05).Compared with IgA1 group and IgA1 +Adv-EGFP group,MTT absorbency was obviously reduced after incubation with aIgA1 for 24 h and 48 h in IgA+Adv-SOCS3-IRES2-EGFP group,and the protein and mRNA expression of TLR4 and TGF-β1 were significantly decreased in IgA1 +AdvSOCS3-EGFP group (P＜ 0.05).Conclusion Over-expression of SOCS3 may inhibit the proliferation of HMC stimulated by aIgA1,partly through down-regulating the expression of TLR4 and TGF-β1.