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.

Objective To study the expression of nephrin, podocin and ? actinin in glomeruli in puromycin aminonucleoside (PAN) nephrosis rat,and to disclose the possible association of these molecules with the development of proteinuria and the relation among these molecules′changes. Methods PAN nephrosis rat models were established by a single intraperitoneal injection of PAN. Indirect immunofluorescence (IF) staining and real time quantitative reverse PCR were used to study the expression of nephrin, podocin and ? actinin in glomeruli of model rats at 12 hours, 1 day, 36 hours,2 days, 5 days, 10 days, 15 days and 20 days after PAN injection. Results (1)In PAN rats, the urinary protein reached the peak level (P=0 02) at the 10th day and decreased back to control level at the 20th day. (2)The IF intensity of podocin decreased significantly at the 36th hr (P=0 04), the 2nd day(P=0 03), the 5th day(P=0 04) and the 10th day(P=0 006),while at the 15th day,the intensity began to recover (P=0 007)and reached to the control level at the 20th day. The distribution of podocin showed a linear pattern along the glomerular capillary wall (GCW) in control rats and discontinuous in PAN rats. (3)The intensity of nephrin staining decreased significantly at the 5th day (P=0 002), 10th day (P=0 007) and began to recover at the 15th day (P=0 04), while still abnormal at the 20th day(P=0 02). The distribution of nephrin in control rats showed a linear pattern along the GCW whereas a discontinuous pattern in PAN rats from the first day throughout the course. (4)The intensity of ? actinin in PAN rats increased significantly at the 20th day(P=0 009) accompanied by the increase of area ratio (P=0 007). (5)The mRNA of nephrin decreased(P =0 02)at the 5th day and recover at the 10th day. Conclusions Nephrin and podocin change prior to the presence of heavy proteinuria, which suggests their causative effects to proteinuria in PAN nephrosis. The distribution changes of nephrin and podocin occurre before the decrease of their expression level and seem to be an initial trigger factor of proteinuria. The significant decrease and recovery of podocin presents earlier than those of nephrin.The proteinuria level is correlated with the expression of podocin and nephrin.

Objective: To investigate the association between nephrin, podocin and ? actinin of the glomerular podocyte molecules, the morphometric change of podocyte foot process and the development of proteinuria. Methods: Puromycin aminonucleoside (PAN) nephrosis was established. Immunofluorescence staining, image analysis and real time quantitative PCR were employed to study the distribution and quantitation of glomerular expression of nephrin, podocin and ? actinin. Morphometric methods were applied to evaluate the morphology change of podocyte foot processes under electron microscopy. Results: (1) Before the onset of proteinuria, 2 days after PAN injection, the podocyte foot process became swollen;nephrin and podocin staining were changed into discontinuous pattern accompanied by the decrease of podocin staining intensity. The foot process became more swollen on day 5,and podocin intensity continued to decrease. Meanwhile, nephrin decreased significantly both in protein intensity and at mRNA level. (2) When heavy proteinuria [(130.8?30.7) mg/d, P =0.02]occurred, complete effacement of podocyte foot processes was revealed; both podocin and nephrin staining intensity decreased dramatically( P

Abstract： Arsenic and arsenic compounds have been listed as one of the toxic and harmful environment pollutants, and drinking, seafood intake, use of skincare products and inhalation of tobacco smoke are main routes of exposure to human arsenic exposure. The adverse effects of arsenic on pregnant outcomes have been paid much attention. Prenatal exposure to high-level arsenic has been found to increase the risk of spontaneous abortion among pregnant women. Based on national and international epidemiological studies on the correlation between arsenic exposure and spontaneous abortion during the period between 1992 and 2020, we review the association between arsenic exposure and spontaneous abortion and describe the mechanisms underlying spontaneous abortion caused by arsenic exposure, so as to provide insights into early prevention of spontaneous abortion.

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.

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

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.

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.