Objective To explore the pertinence between the concentrations of seminal leptin (lep) and genital endocrine hormones, such as testosterone(T), follicle stimulating hormone( FSH), luteotropic hormone(LH) and insulin growth factor-1 (IGF-1), as well as the effect of Lep on sperm concentration,motility and genital function indexes.Methods 126 cases of infertility and 30 cases of normal fertility were randomly chosen.The contents of Lep, T, FSH and LH were detected by radioimmunoassay (RIA) and the IGF-1 content were detected by immunoradiometric assay (IRMA).The infertility group was divided into group A (sperm count ≥ 20×109 /L), group B (sperm count ＜ 20×109 /L) and group C of azoospermia.According to the number of white blood cell ( WBC ) in 10 high power fields (HPF), the infertility group was further divided into group WBC (WBC count ≥ 1×109 /L) and group Non-WBC(WBC count ＜ 1×109 /L).According to the sperm vitality and the rate of motility, group A was subdivided into the group of normal spermatic vitality (a + b≥50% ), the group of abnormal vitality( a + b ＜ 50% ) (a refers to the number of sperms with fast forward movement, and b refers to the number of sperms with slow or sluggish forward movement), the group of normal rate of spermatic motility( the rate ≥60% ) and the group of decreased rate of motility( the rate ＜60% ).According to the examination results of the normal contrast group, group A and group B were respectively divided into the following groups: the group of normal sperm penetrating power(≥40 mm), the group of decreased penetrating power( ＜ 40 mm), the group of normal intact acrosome rate(≥80% ), the group of decreased intact acrosome rate( ＜ 80% ), the group of normal terminal swelling rate( ≥60% ) and the group of decreased terminal swelling rate( ＜60% ).Results The concentration of Lep in the infertility group was (2.77±0.80) μg/L, significantly higher than the level of Lep [ (1.14 ± 0.31 ) μg/L ] in the contrast group ( t = 10.943,P ＜ 0.05 ).The contents of IGF-1 and T were ( 17.67±8.09) μg/L and (4.84±2.15) nmol/L respectively, significantly lower than the levels of IGF-1 [(24.79±9.32) μg/L] and T [(6.30±2.53) nmol/L] in the contrast group (t =4.205,3.228,P＜0.01).There existed no significant differences of the concentrations of FSH and LH between the two groups ( t = 1.655,1.378 ,P ＞ 0.05 ).The concentrations of FSH and LH were (32.61±9.14) U/L and (40.57 ± 12.40) U/L respectively in the infertility group and(29.63±7.56) U/L and (37.25±9.19) U/L respectively in the contrast group.The concentrations of Lep and IGF-1 and T showed negative correlation in the infertility group (r = -0.237, -0.316,P ＜ 0.01 ).The concentration of Lep had no correlation to the FSH and LH concentration (r = 0.104, 0.112, P ＞ 0.05 ).The concentration of Lep showed a gradual increase within group A, group B and group C (F = 115.93, P ＜ 0.01 ).The leptin contents in the above mentioned normal subgroups were found to be lower than the abnormal groups.Conclusions There exists pertinence between the Lep concentration and the concentrations of TGF-1, T, FSH and LH.Lep may decrease the sperm density and inhibit sperm vitality and the motility rate through inhibiting androgen secretion and sperm capacitation.

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 .

To study the effect of urokinase on platelet activation and aggregation in the patients with glomerulonephritis. Platelet counts(PC),mean platelet volume (MPV), and platelet distribution width (PDW) were determined with automatic blood cell analytic apparatus, the expressive levels of platelet granular membrane protein (GMP 140) and fibrinogen receptor(GPⅡb Ⅲa) on the surfaces of platelets were assayed with flow cytometry, and the levels of fibinogen and fibrin degradation products (FDP) in blood were measured with ELISA in 102 children with glomerulonephritis. The results showed that urokinase led to PC decreas,MPV increase and PDW exceeding normal value in 102 children with glomerulonephritis, including micro change (MC),glomerulosclerosis (GS),membranous nephropathy (MN), and proliferative glomerulonephritis (PGN).The expressive levels of GMP 140 after the treatment with urokinase were significantly higher than that before the treatment and controls( P 0 01). The results suggested that urokinase used in the patients with glomerulonephritis increased platelet activation function and decreased platelet aggregation function.

In order to improve the diagnosis and treatment of severe hemolytic uremic syndrome (SHUS),we summarized the rescuing experiences in 3 children with SHUS.Therefore,close observation of gastro intestinal symptoms,collaborated with repeated examinations of the blood,renal function,urinary output,blood pressure,and manifestations of the central nervous system could help predict to certain extent the occurrence of further hemolysis and renal failure in these patients.Energetic treatment of malignant hypertension and relieving the symptoms of center nervous system could successfully improve renal functions.If urinary output decreased quickly,creatinine (Cr) and blood urea nitrogen (BUN) increased,or severe infection occurred in SHUS patients,hemodialysis should be undertaken as early as possible to prevent caute renal failure and multiple organ failure syndrome (MOFS).Therefore,the treatment of SHUS in children has its peculiarity and complexity.

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.

0.05).Conclusion VAMT is as effective as VATS,whereas it can achieve a shorter operation time.

Objective To investigate the clinical characteristics and classification of diabetic patients with unprovoked ketoacidosis.Methods According to body mass index(BMI),56 patients were divided into 3 groups: low body weight group(LBW,BMI25 kg/m2,n=20).Clinical characteristics,including age,gender,the course of the disease,positive rate of glutamic acid decarboxylase antibody(GAD-Ab) and the function of islet beta-cell,were compared between these three group.Results There were no significant differences in some clinical features(glycemia,and ketosis status) at beginning of disease between 3 groups.The level of serum TG and Insulin was higher in OBW group than that in other groups.Only 3 patients were found GAD-Ab positive in LBW group.Conclusion The clinical and immunological characteristics of OBW and LBW patients were quite different,some of the obese patients should be classified into type2 diabetes,and some into idiopathic type1 diabetes.

Objective To investigate the regulatory effects of umbilical cord-derived mesenchymal stem cells (UC -MSCs) on Th17 cells and related cytokines in patients with systemic lupus erythematosus (SLE). Methods Human UC-MSCs were isolated and expanded and infused into fourteen SLE patients. Clinical changes were evaluated before and after transplantation by SLE disease activity index (SLEDAI), 24-hour urine protein, serum albumin and complement C3. The percentages of CD3 +CD8-IL17A + T cells in peripheral blood were detected by flow cytometry. Concentrations of plasma IL-6, TGF-β, IL-17A, IL-22were determined by enzyme-linked immunosorbent assay (ELISA). UC-MSCs and peripheral blood mononindependent samples t-test. Results SLEDAI scores decreased significantly at 3 month (7.8±1.2, t=2.19) and 6 month (6.9±0.9, t=4.2) after UC-MSCs transplantation than pre-transplantation level (10.4±0.9, P< 0.05). Twenty-four-hour proteinuria decreased significantly 6 months after MSCs infusion [(1489±260) mg vs (2454±322) mg, t=2.6, P<0.05]. Meanwhile, serum albumin and complement C3 levels had increased significantly since 1 month after transplantation (P<0.05). The percentages of peripheral blood CD3+CD8-IL17A+T cells decreased obviously in 1 week, 1 month and 6 months after UC-MSCs transplantation (all P<0.05). The coculture of UC-MSCs with PBMC from SLE patients resulted in a statistically significant reduction of CD3+CD8-IL17A+T cells percentage in PBMC (P<0.05), but was not in a dose dependent manner. No change of plasma IL+6, TGF-β, IL-17A and IL-22 levels was observed after UC-MSCs transplantation (P>0.05).Conclusion UC-MSCs transplantation down-regulates the percentages of CD3+CD8-IL17A+T cells in SLE patients, which may be one of the mechanisms for its therapeutic effect in refractory SLE.

Objective To construct a eukaryotic expressing vector harboring human DC-SIGN, and establish a BHK21 cell line stably and highly expressing DC-SIGN. Methods The DC-SIGN gene fragment which contained Not I and BamH I sites was amplified by PCR from pUNO-hDCSIGN1Aa plasmid, digested with Not I and BamH I, and then cloned into an eukaryotic expression vector pIRES-neo to construct eukaryotic expression vector pIRES-neo-DC-SIGN. The recombined plasmid was identified with Not I and BamH I enzyme digestion and sequencing, the latter was then transfected to BHK21 cells by LipofectamineTM 2000. After screening culture by G418, BHK21 cell line stably expressing DC-SIGN was established. The expression of DC-SIGN was identified by flow cytometry, Western blotting and immunofluorescence method. Results The gene sequence of DC-SIGN was consistent with that of design. PCR and double enzyme digestion analysis showed that the recombinant plasmid pIRES-neo-DC-SIGN was constructed successfully. After transfection, positive clones were selected with G418. After limiting dilution assay, BKH21 cell lines stably expressing DC-SIGN were established. The detection result of flow cytometry showed that the expression ratio of DC-SIGN positive clones was close to 90%. The result of immunofluorescence displayed that the expression of DC-SIGN was mostly located on the surface of cell membrane. Western blotting displayed the specific band of DC-SIGN protein. It showed that the BHK21 cells stably expressing DC-SIGN were successfully established. Conclusion DC-SIGN eukaryotic expression vector has been successfully constructed. The successful establishment of BHK21 cell lines which can stably express DC-SIGN provides a substantial foundation for further study on the DC targeting vaccines.

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.