The protein expression of cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-activated Cl(-) channel, in ovarian stimulated premature female rat ovary during a cycle of follicle development and corpus luteum formation was investigated. Animals were injected with 10 U pregnant Mare's serum gonadotropin (PMSG) and subsequently 10 U hCG 48 h later. Time-dependent immunohistochemistry and Western blotting experiments were performed before and 24, 48, 72 h after hCG treatment. The immunohistochemistry revealed that administration of PMSG stimulated the CFTR expression in thecal cell layer and granulosa cell layer of mature follicles 48 h post injection, coincident with the PMSG-induced peak in follicular estradiol. However, the expression of CFTR in the granulose lutein cell layer and thecal lutein cell layer was time-dependently reduced following hCG injection, in accordance with the gradually increased progestogen level during luteum corpus formation. Western blotting analysis demonstrated that rat ovarian tissue expressed the special CFTR band at 170 kD. It is concluded that cAMP-dependent Cl(-) channels are involved in regulation of follicle development and luteum formation.
Connective Tissue Growth Factor/genetics ; Connective Tissue Growth Factor/*metabolism ; Connective Tissue Growth Factor/*physiology ; Corpus Luteum/growth & development ; Ovarian Follicle/growth & development ; Ovary/*metabolism ; Rats, Wistar

The CrdS protein responding to the acidic adaptation was prokaryotic-expressed in our Laboratory to explore the regulatory mechanism in the acidic adaptation of Helicobacter pylori (H. pylori). The whole genomic DNA of H. pylori strain 26695 was abstracted and set as the template firstly. And then the hp1364 gene coding CrdS protein was amplified via the PCR technique. Then the clonal recombinant plasmid pUCm-T-hp1364 and the prokaryotic expression plasmid pQE30-hp1364 were built and identified by the methods of PCR, cutting with two enzymes and sequencing. After that, the plasmid pQE30-hp1364 was transferred into the E. coli XL1 blue and induced with IPTG. Using western blot and SDS-PAGE, it can be analyzed that the expressed recombinant protein existed mainly in the form of the inclusion bodies and its relative molecular mass was about 46 kDa. The successfully attained recombinant protein CrdS will provide the material to explore the regulatory mechanism in the acidic adaptation of H. pylori and the new way to resist the infection of H. pylori.
Bacterial Proteins ; biosynthesis ; genetics ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; metabolism ; Helicobacter pylori ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; genetics

ObjectiveTo study the effect of lentivirus-mediated microRNA-21 RNAi on biological behaviors in human hepatic cancer cells.MethodsMicroRNA-21 specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector,pGCSIL-GFP.HepG2 cells were infected by microRNA-21 siRNA recombinant lentivirus (miR-21-siRNA-Ⅳ).The HepG2 cells were devided into SI group,NC group and N group in vitro.The expression of the targets of miR-21 was detected by RT-PCR.Cell growth was analyzed by MTT assay.The invasion was dectected by tmnswell method.Apoptosis was detected by Hoechst33258.BALB/c nude mice were randomly divided into SI group and NC group.The growth of transplant tumors in BALB/c nude mice were observed.Results ( 1 ) The expression level of miR-21 was inhibited significantly by miR-21-siRNA-lⅣ.(2) The proliferation of HepG2 was also markedly suppressed in MTT at the 96 h point ( P =0.0031,P ＜ 0.05 ).(3) The number of cells that migrated through the chamber of SI group decreased ( P =0.0004,P ＜ 0.05 ).(4) The cell apopotosis in SI group increased markedly.In addition,the caspase 3 mRNA significantly increased ( P =0.0002,P ＜ 0.05 ).( 5 ) Tumor growth curve was not statistically different between groups ( P =0.0002,P ＜ 0.05 ).ConclusionsMicroRNA-21 specific siRNA suppresses the proliferation and migration of HepG2 cells and induces tumor cell apoptosis inhibiting the growth of transplanted tumor in Balb/c nude mice.

0.05],significantly higher than stage Ⅲ,Ⅳ[60%(6/10),50%(5/10);P

Objective To explore the correlation between Helicobacter pylori (Hp) infection and recurrent abdominal pain (RAP) in children over 6 years old and its relative treatment.Me-thods One hundred and eighty children over 6 years old with the diagnosis of RAP from Mar.2007 to Feb.2009 were selected,30 healthy children without RAP were taken as the healthy control group at the same period.14C-urea breath test (14C-UBT) was used to detect whether the patient was infected by Hp.The radical cure of Hp was given to the Hp-positive children with RAP,and the remission rate of children with RAP and the Hp negative rate were observed.Results The positive rate in RAP group was 58.33%,which was higher significantly than that in healthy control group(20.00%)(P0.05).The positive rate of those with and without bad eating habits were 45.56% and 12.78%,and the positive rate of with and without family gastrosis history was 49.44% and 8.89%,which both had significant difference between them(Pa

Objective To isolate and culture sweat gland epithelial cells in vitro,and to study the effects of acetylcholine (ACh) on intracellular flee calcium concentration ([Ca~(2+)]i) of cultured sweat gland epithelial cells.Methods Sweat glands epithelial cells were collected by enzymatic digestion.After ACh was added to the primary and first passage cells,[Ca~(2+)]i was examined using confocal laser scanning microscopy (CLSM) and the Ca~(2+) sensitive dye Fura 3/AM.Results The primary and first passage epithe- lial cells grew well.After ACh was added,opening of the calcium channel and significant [Ca~(2+)]i increase were observed when the primary and first passage cells were incubated with high concentration of calcium (2 mmol/L);no significant [Ca~(2+)]i increase was observed in those cultured without calcium.Conclusion Upon stimulation with ACh,calcium channels of cultured primary and first passage sweat gland epithelial cells would open,influx of extracellular Ca~(2+) occurred,which resulted in an increase of [Ca~(2+)]i.Extracellular bound calcium was therefore converted into intracellular free calcium.

Objective To investigate the diagnosis and treatment of anticoagulation complication,heparin-induced thrombocytopenia (HIT),induced by low-molecular-weight heparin (LMWH) after joint replacement.Methods Retrospectively analyzed the clinical manifestations,treatment and prognosis of 4 patients after total knee arthroplasty (TKA) suffered from HIT induced by LMWH.Results The morbidity rate of HIT induced by LMWH was 0.3％(4/1376).Platelets were found less than 70 × 109/L after 4-9 d,minimum to 16 × 109/L.Different level of cardiorespiratory distress and lower extremity deep venous thrombosis all appeared.Once diagnosed,patients were immediately stopped using LMWH and replaced with argatroban,fondaparinux or rivaroxaban and warfarin.Deep vein thrombosis gradually disappeared and cured in 3 patients after treatment.One patient suffered cerebral hemorrhage and multiple pulmonary embolism,received craniotomy hematoma removal and decompressive craniectomy.But the patient didn't wake up after 1-month treatment,became a vegetative one.Conclusions HIT is rare but dangerous,which can result in deep vein thrombosis,pulmonary embolism,cerebral hemorrhage and other serious consequences.Early diagnosis and timely treatment can reduce the rate of disability and mortality.

Female ; Hemangioma ; microbiology ; therapy ; Humans ; Infant ; Infant, Newborn ; Male ; Microbial Sensitivity Tests ; Ulcer ; microbiology ; therapy

Objective:To investigate the feasibility and safety of perimembranous ventricular septal defects (PmVSD) closure solely by femoral vein approach under transesophageal echocardiography (TEE) guidance.Methods:From January 1,2014 to May 31,2016,26 patients with PmVSD in Second Xiangya Hospital were selected,with age at 3.2-6.0 (4.3±0.7) years old and body weight at 15.0-19.5 (16.7±1.4) kg.The diameter of VSD was 3.5-4.8 (4.1±0.3) mm.All patients were treated by percutaneous PmVSD closure solely by femoral vein approach under TEE guidance.The effect of the procedure was evaluated by TEE and transthoracic echocardiography (TTE).The clinical follow-up study was conducted by TTE at 1,3,6 and 12 month (s) after the procedure.Results:Twenty cases were successfully treated with percutaneous PmVSD closure solely by femoral vein approach under TEE guidance,and the success rate was 76.9％.Six patients were converted to perventricular closure under TEE guidance because the guide wire in two cases or catheter in other cases could not pass through PmVSD.The diameter of symmetrical VSD occluder was 6.0-7.0 (6.2±0.4) mm.The procedural time was 12.0-64.0 (26.8±6.3) min.The residence time at ICU was 1.8-2.4 (26.8±6.3) h.The in-hospital time was 4.0-5.0 (4.4±0.5) d.There were 3 patients with immediate post-operative trivial residual shunt and incomplete right bundle branch block (IRBBB).All patients survived with no peripheral vascular injury or complications such as tricuspid regurgitation,pericardial tamponade and pulmonary infection.The residual shunt disappeared in 3 patients and IRBBB became normal rhythm in 3 patients at 1 month follow-up time point.No patients suffered from occluder malposition,residual shunt,pericardial effusion,arrhythmia (atrio-ventricular block),aortic valve regurgitation and tricuspid regurgitation.Conclusion:TEE-guided percutaneous PmVSD closureby femoral vein approach is safe and effective.

Objective To observe the symptom and characteristics of liver injury induced by different dosage of long-term administration of rifampicin(RIF) in mice.Methods Twenty-four healthy female ICR mice were randomly divided into 4 groups(6 mice in each group):control group,low dosage group,medium dosage group and high dosage group.The four groups were treated with 0,100,200,400 mg·kg-1·d-1 RIF respectively for 2 weeks.Mice blood and liver tissue samples were collected at 6 hours after the last administration for serological test and liver histological observation.Results No mice died before execution.The TBA,DBIL and TBIL of high dosage group all increased compared with the control group, but the ALT,AST and ALP showed no obvious change.The TBA and DBIL of medium dosage group increased compared with the control group, while the TBIL,ALT,AST and ALP showed no obvious change.In the low dosage group,there was no obvious change in terms of TBA,DBIL,TBIL,ALT,AST,and ALP compared with the control group.Obvious pathological change occured in the liver of mice in all the experimental groups.HE staining showed edema and feather steatosis in liver cells, accompanied by a large number of inflammatory cells infiltration and a few sporadic cholestasis.With the increasing of RIF dosage,the liver pathological change became more obviously.In the experimental group,electron microscope showed that there were a lot of fat droplets in the liver cells wrapped slurry,and part of the capillary bile duct were slightly expanded with different electron density and irregular shape of bile sample material inside.The pathologic changes get more obvious with the increase of the concentration of rifampicin as well.Conclusion RIF could induce liver injury after 2 weeks' treatment at different dosages,mainly pathological changes included liver cell steatosis,inflammatory cell infiltration,and cholestasis.Rifampicin induced liver injury in a concentration-dependent manner.However,the mechanism of rifampicin-induced liver injury in mice needs further study.