Objective To explore whether the inhibited expression of fibrocystin by RNA interference can increase epidermal growth factor (EGF)-induced cell proliferation and its possible mechanism . Methods A stable PKHD1-silenced HEK 293 cell line was established . Cell proliferation rate, intracellular Ca2+ concentration and extracellular signal-reguhted kinase 1/2(ERK1/2) activity were assessed after treatment with EGF, verapamil and Bay K8644 . Results The proliferation rate of PKHD1-silenced HEK-293 cells was found to be significantly higher after EGF stimulation compared to the control HEK 293 cell (231 .5% vs 152 .8%, P＜0 .01) . PKHD1-silencing lowered the intracellular Ca2+ concentration and caused EGF-induced ERK1/2 overactivation in the cells(P＜0 .01 ) . When cells were treated with verapamil for 4 hours to lower the intracellular Ca2+ concentration, the cell proliferation rate was significantly increased after 20 ng EGF for 24 hours . The verapamil treatment increased the level of activated ERK1/2 in EGF-treated cells . An increase of intracellular Ca2 + in PKHD1-silenced ceils repressed the EGF-dependent ERK1/2 activation and the hyperproliferative response to EGF stimulation . Conclusions Inhibition of fibrocystin can cause EGF-induced excessive proliferation through decreasing intracellular Ca2+ resulting in EGF-induced ERK1/2 activation . The loss of fibrocystin may lead to abnormal proliferation in kidney epithelial cells and cyst formation in ARPKD through modulation of intracellular Ca2+ concentration .

Objective:To study the possible therapeutic mechanisms of triptergium wilfordi in treating glomerulonephritis.Methods:Thirty seven patients with IgA nephropathy were detected urine mononeuclear cells densities with flow cytometry during taking triptergium wilfordi tablets,30 healthy people were used as normal control.Results:The patients urine CD3 +?CD4 +?CD8 +?CD14 + and CD44 + cells densities were much higher than normal control's,but obviously decreased after triptergium wilfordi treatment.CD4/CD8 ratio was much lower in patients than in normal control.It increased markedly after treatment.Patients who reached remission initially had a lower CD4 + cell percentage and higher CD14 + cells percentage than those showed no response to triptergium wilfordi treatment.Conclusion:Triptergium wilfordi could modulate kidney immune cell function and adhesive molecule CD44 expression.Its effects were associated with renal immunity.

Platelet count (PC) and plasma prothrombin time (PPT) of sixty- six patients with cyanotic congenital heart disease (cyanotic CHD) were determined, and linear relationship analysis was made between PC, PPT and hemoglobin (HB), Hemotocrit saturation of blood (SaO2). Blood viscosity (BV) and hemotocrit (HT) were also examined. It was discovered that there was a significant positive linear relationship between SaO2 and PC, HB and PPT, and a significant negative linear relationship between PC and HB, BV, HT, respectively. There was a significant difference of PC in cyanotic-CHD (n = 66) as compared with noncyanotic-CHD (t=13.9508, P

Objective To analyze the alterative characteristics of the extravascular lung water (EVLW ) in the patients with septic shock and clarify its value on the prognosis of these patients.Methods By the methods of retrospective analysis,according to the ultimate survival,21 patients with septic shock were divided into survivor group (10 cases) and non-survivor group (11 cases).The clinical features of the patients were observed and hemodynamic monitoring was made with PiCCO monitor.The EVLW was measured and the relationship between the EVLW and the prognosis of patients was analyzed.Results On the first,second and third day,EVLW was (12.7 ±1.8),(11.3 ±1.3),(10.1 ±1.3) ml/kg in survivor group,and (14.4 ± 1.0),(14.6 ± 1.4),(14.6 ±1.3) ml/kg in non-survivor group respectively,and there were statistical differences between two groups (P <0.05).However,on the second day after the intensive therapy,EVLW in survivor group dropped significantly(P＜0.05),but the non-survivor group only declined slightly,and compared with the result of the first day,there was no obvious difference (P >0.05).Conclusions The EVLW in the patients with septic shock increases significantly.The dynamic changes of the EVLW may be one of the factors for predicting the prognosis of patients with septic shock.

ObjectiveTo evaluate the effect of xuebijing injection on inflammatory response and cellular immune function in patients with severe sepsis.MethodsSixty-two patients with severe sepsis from September 2008 to August 2009 were divided into treatment group(30 patients) and control group (32patients) by random digits table.All the patients received sepsis-bundle therapy and patients in treatment group added xuebijing injection therapy with 100 ml,twice a day,for 7 days.The levels of serum tumor necrosis factor-alpha(TNF-α ),interleukin(IL)-6,IL-10 and C-reactive protein (CRP),peripheral blood T lymphocyte CD4+、CD8+ 、CD4+/CD8+,the expression of human leucocyte antigen (HLA)-DR on CD14+peripheral blood mononuclear cell (PBMC) were detected before and aftertreatment.ResultsThere was no significant difference in the levels of serum TNF-o,IL-6,IL-10,CRP,peripheral blood Tlymphocyte CD4+、CD8+、CD4+/CD8+ and the expression of HLA-DR on CD14+ PBMC before treatment between two groups(P ＞ 0.05).After treatment,compared with those in control group,the levels of serum TNF- α,IL-6,IL-10 and CRP in treatment group were significantly decreased [ ( 64.4 ± 13.5) ng/L vs.(96.1 ± 22.1 ) ng/L,( 153.8 ± 23.8 ) ng/L vs.(180.1 ± 21.7) ng/L,(73.8 ± 13.8) ng/L vs.(101.1 ± 11.7) ng/L,(53.7 ± 18.8) mg/L vs.(91.3 ± 32.8)mg/L,P ＜0.05],while peripheral blood T lymphocyte CD4+/CD8+ and the expression of HLA-DR on CD14+PBMC were significantly increased [ 0.311 ± 0.021 vs.0.424 ± 0.035,0.201 ± 0.017 vs.0.238 ± 0.038,1.78 ±0.21 vs.1.56 ±0.18,(38.4 ± 11.5)％ vs.(18.1 ± 12.1)％,P＜0.05].ConclusionXuebijing injection can reduce the inflammatory response and ameliorate immune disorder in patients with severe sepsis.

Objective To investigate and compare the change of extravascular lung water (EVLW) and levels of cytokines in septic patients without clinical acute lung injury (ALI) /acute respiratory distress syndrome (ARDS) with those in spetic patients with sepsis-induced ALI/ARDS in order to determine the role of EVLW involved in the pathogenesis of lung injury in the patients by quantifying the relationship between EVLW and biomarkers of lung injury in patients with sepsis.Methods A total of 40 septic patients complicated either with or without clinical ALI/ARDS after sepsis. In each patient,transpulmonary thermodilution (PiCCO) was used to measure cardiovascular hemodynamics and EVLWI for 7 days via an arterial cannula indwelled within 72 hours after diagnosis of severe sepsis was made, and serial bronchoalveolar lavages (BAL) were carried out.Other examinations including blood gas analysis,ventilator parameters,chest X-ray and the levels of pro-inflammatory cytokines such as tumor necrosis factor (TNF-oα),interleukin-1 in the BAL were recorded.In-hospital and ICU mortalities were also observed.Results Of total 40 patients,29 were complicated with clinically defined septic ALI/ARDS ( ARDS n =15,and ALI n =14).The septic patients complicated with ALL/ARDS had significantly higher amount of EVLWI and higher levels of TNF-α and interleukin-1 in the BAL than patients without ALI/ARDS ( P ＜ 0.05).The arterial oxygen tension/fractional inspired oxygen ratio,lung injury score,and the levels of TNF-α,IL-1 in the BAL correlated with EVLWI.Moreover,in-hospital mortality,ICU mortality and the length of ICU stay of the patients with high amount of EVLWI were markedly increased than those of patients with low amount of EVLWI. Conclusions In septic patients complicated with ALI/ARDS, the extravascular lung water index correlates with oxygenation,lung injury severity and inflammatory cytokines in lung.Determination of EVLWI may be useful for evaluation of severity of lung injury and prognosis of septic patients.

AIM: To investigate the effects of Lipopolysaccharide(LPS) and interieukin 1 receptor antagonist (IL- 1ra) on mesangial cells proliferation and nitric oxide synthesis. METHODS: Glomerular mesangial cells from SD rats were cultured. The first and second passages of cultured cells were used for the experiment. LPS and LPS plus IL- 1ra were added in cell cultures, respectively. By using chemical method the nitrite in supernatants was measured ,3H- TaR incorporation was determined to evaluate the GMC proliferation. Northern and slot hybridizations were performed to detect the expression of iNOS mRNA. RESULTS: There were expression of iNOS mRNA, more production of nitrite(0.64 + 0.25 vs 0. 12 + 0.06 nmol/104 cell) in supernatants and GMC proliferation(3735 + 1177.9 vs 1785 + 280.6) in LPS group compared to the control. While compared with LPS group, in LPS + IL- 1ra GMC group, expression of iNOS mRNA decreased by 40%, nitrite increased(3.28 + 0.33 nmol/104 cell), proliferation of GMC decreased (818 + 77.27). CONCLUSION: LPS could activate the GMC to express iNOS mRNA and produce more nitrite. IL - 1ra could partially inhibit the effects of LPS on the expression of iNOS mRNA in GMC, but not nitrite. There is no synchronous correlation between NO production and GMC proliferation.

G) was relatively rare. Conclusion: DHPLC is a rapid and reliable method for polymorphism screening. Eight polymorphisms in the NR3C1 gene are detected in Chinese population with the technique of DHPLC , of which two haplotypes have been identified for the first time.

Objective To analyze type IV collagen a5 chain mRNA and to study the relationship between genotype and phenotype in X-linked Alport syndrome. Methods Total RNA was isolated from the cultured skin fibroblasts of 21 unrelated Chinese X-linked Alport syndrome patients (17 males and 4 females),then ?5 chain (Ⅳ) mRNA was analyzed by using reverse-transcription-polymerase reaction (RT-PCR) and direct sequencing. Meanwhile,by using PCR and direct sequencing,detection of COL4A5 gene mutations at genomic DNA level was carried out. Results Abnormal ?5 chain IV cDNA was detected in 21 X-linked Alport syndrome patients,and seventeen mutations detected in this study were novel mutations. In 15/21 of patients,identical COL4A5 mutations were detected both at mRNA level and genomic DNA level,and in 6/21 of patients with splicing-site mutations,changes in transcript structure differed from changes in genomic DNA level. 16/21 of patients belonged to X-linked juvenile kindreds,and 2/21 of patients to adult kindreds. Conclusion Different type of mutations in COL4A5 can lead to the severe form of X-linked Alport syndrome,and mRNA-based procedures can both directly detect mutations in the coding sequences,as well as changes in transcript level or structure,and can identify some abnormalities that would otherwise have been missed by DNA-based procedures.

AIM: To investigate the effects of Lipopolysaccharide(LPS) and interleukin 1 receptor antagonist(IL-1ra) on mesangial cells proliferation and nitric oxide synthesis. METHODS: Glomerular mesangial cells from SD rats were cultured. The first and second passages of cultured cells were used for the experiment. LPS and LPS plus IL-1ra were added in cell cultures, respectively. By using chemical method the nitrite in supernatants was measured,3H-TdR incorporation was determined to evaluate the GMC proliferation. Northern and slot hybridizations were performed to detect the expression of iNOS mRNA. RESULTS: There were expression of iNOS mRNA, more production of nitrite(0.64?0.25 vs 0.12?0.06 nmol/104 cell) in supernatants and GMC proliferation(3735?1177.9 vs 1785?280.6) in LPS group compared to the control. While compared with LPS group, in LPS+IL-1ra GMC group, expression of iNOS mRNA decreased by 40%, nitrite increased(3.28?0.33 nmol/104 cell), proliferation of GMC decreased (818?77.27). CONCLUSION: LPS could activate the GMC to express iNOS mRNA and produce more nitrite. IL-1ra could partially inhibit the effects of LPS on the expression of iNOS mRNA in GMC, but not nitrite. There is no synchronous correlation between NO production and GMC proliferation.