Actins ; metabolism ; Animals ; Collagen Type III ; metabolism ; Collagen Type IV ; metabolism ; Desmin ; metabolism ; Diabetes Mellitus, Experimental ; complications ; Diabetic Nephropathies ; metabolism ; pathology ; HSP47 Heat-Shock Proteins ; metabolism ; Kidney Glomerulus ; metabolism ; pathology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Vimentin ; metabolism

OBJECTIVETo study the feasibility and risk of endoscopic retrograde cholangiopancreatography (ERCP) for children in clinic, and to evaluate the effects of ERCP in diagnosis and treatment of the pancreatic diseases in children.

METHODTotally 98 patients under 14 years of age who underwent ERCP from 1994 to 2011 were enrolled in the study. The data of diagnosis, anesthesia type, treatments, and postoperative complications were collected.

RESULTThe 98 patients were 4 to 14 years old, of whom 32 cases suffered from acute pancreatitis (30 cases with biliary disease and 2 with hyperlipidemia); 42 cases had chronic pancreatitis, of whom 36 had calculus of pancreatic duct, pancreatic pseudocyst was seen in 6 cases. Pancreas divisum was found in 20 cases, choledochopancreatic junction anomaly in 6 cases,and annular pancreas in 1 case. The operations of dissection of pancreatic duct and biliary duct, calculus removal, insertion of endoprosthesis and draining tube in pancreatic duct or biliary duct were performed. No patients died from ERCP complications. In observation of postoperative complications, acute pancreatitis after ERCP occurred in 1 case, with the incidence rate of 1%, 1 case had bleeding (1%), and 5 cases had hyperamylasemia (5%). All the complications were cured within 1 week. The average hospital stay was 5.51 d.

CONCLUSIONERCP is useful and safe in children under suitable condition of doctors and equipments, and no high rates of complications were observed.

Adolescent ; Calculi ; diagnosis ; surgery ; Child ; Child, Preschool ; Cholangiopancreatography, Endoscopic Retrograde ; methods ; Drainage ; Female ; Humans ; Hyperamylasemia ; epidemiology ; etiology ; Magnetic Resonance Imaging ; Male ; Pancreatic Diseases ; diagnosis ; surgery ; Pancreatic Ducts ; diagnostic imaging ; surgery ; Pancreatitis ; diagnosis ; surgery ; Postoperative Complications ; epidemiology ; etiology ; Retrospective Studies ; Sphincterotomy, Endoscopic ; Treatment Outcome

OBJECTIVETo investigate the effect of salidroside on hypoxia-induced apoptosis of corpus cavernosum smooth muscle cells (CCSMCs) in rats.

METHODSRat CCSMCs were cultured in vitro by the enzyme digestion method and identified by immunofluorescent staining of anti-alpha-SMA and anti-Desmin. The non-toxic dose of salidroside was determined by MTT assay. Low-oxygen mixed gas (1% O2, 5% CO2, and 94% N2) was piped into a modular incubator chamber to induce hypoxia. The CCSMCs were divided into a normal, a hypoxia, and a 32 microg/mL salidroside intervention group. The apoptosis of the CCSMCs was detected by flow cytometry and the expression of the caspase-3 protein determined by Western blot.

RESULTSThe majority of the CCSMCs were positive for alpha-SMA and Desmin at immunofluorescent staining. Salidroside at < 32 microg/ml produced no obvious toxicity to CCSMCs. Compared with the normal control group, the rates of early and late apoptosis of CCSMCs were both increased significantly in the hypoxia group ([12.77 +/-1.41]% vs [18.69 +/- 1.29]%, P < 0.01 and [14.63 +/- 2.00]% vs [21.03 +/- 1.530]% , P < 0.05). Western blot showed a markedly increased expression of cleaved caspase-3 (P < 0.01). Intervention with 32 microg/ml salidroside significantly reduced hypoxia-induced early apoptosis of CCSMCs ([13.46% +/- 1.87]%, P < 0.01) and decreased the expression of cleaved caspase-3 (P < 0.01).

CONCLUSIONSalidroside can reduce the expression of cleaved caspase-3 and inhibit hypoxia-induced apoptosis of CCSMCs in rats.

Animals ; Apoptosis ; drug effects ; physiology ; Caspase 3 ; metabolism ; Cell Hypoxia ; physiology ; Cells, Cultured ; Glucosides ; pharmacology ; Humans ; Male ; Myocytes, Smooth Muscle ; cytology ; drug effects ; enzymology ; Penis ; cytology ; drug effects ; Phenols ; pharmacology ; Rats

AIM:To observe the efficacy of intravitreal conbercept injection for chronic central serous chorioretinopathy (CSC).METHODS: Nine eyes of 9 patients diagnosed as chronic CSC between October 2015 to May 2016 were treated with an intravitreal injection of conbercept (0.5mg/0.05mL) (six patients were given the same does of intravitreal injection again at 1mo after the first injection).Follow-up observation was at 1, 2, and 6mo after injection.Observed indicators included best-corrected visual acuity (BCVA), intraocular pressure, optical coherence tomography (OCT), fundus fluorescein angiography (FFA), choroidal indocyanine green angiography (ICGA), macular fovea thickness (CMT), subfoveal choroidal thickness (SFCT).RESULTS:Seven of the 9 patients responded significantly to the drug, while 2 patients had no response.The CMT was 373.12±72.43μm at baseline, which decreased significantly to 332.05±67.13μm, 282.24±62.30μm and 225.56±71.08μm at 1, 2 and 6mo after the intravitreal injection.The mean thickness of SFCT was 422.11±64.82μm before treatment.The choroidal thickness of non-responsive patients before treatment was below average, respectively 353μm and 365μm.The SFCT of 1, 2, and 6mo after treatment was 391.45±75.24μm, 365.53±63.07μm, 355.40±66.65μm.Before treatment and 1mo after, there was no significant difference (P=0.074), but there was statistically significant (P<0.01) between those of before and 2mo and 6mo after.The mean BCVA of the prior treatment was 0.53±0.32, the after treatment was 0.65±0.20, there was no different between the two(P>0.05).CONCLUSION: Intravitreal conbercept injection in chronic CSC may have some effect in accelerating subertinal fluid resolution and decreasing the CMT.The SFCT within 6mo after treatment was significantly lower than pretreatment.The SFCT may be an indicator of whether patients respond.

Objective To investigate independent prognostic factors for overall survival (OS) in extensive disease small cell lung cancer (EDSCLC). Methods Between January 2003 and December 2006, 154 patients diagnosed with extensive stage small cell lung cancer were enrolled in this study.Prognostic factors such as gender, age, performance status, smoking history, weight loss, distant metastasis, the number of matastasis, brain metastasis, the cycle of chemotherapy and thoracic radiation therapy (TRT) for EDSCLC patients were evaluated by univariate and multivariate analysis. Results The median following-up time was 40. 5 months. The rate of follow-up was 92. 2%. The MST and overall survival rates at 3-year in smoking group and no-smoking group were 13 months, 11.8% and 17 months,22. 8%,respectively (χ2=3.40,P =0. 064);in ChT/TRT group and ChT group, they were 17. 2 months, 17.9%and 9.3 months,13.9%, respectively(χ2=10.47,P=0.001);and in the cycle of chemotherapy ≥4 group and ＜ 4 group, they were 16 months, 20. 1% and 9.3 months, 2. 9%, respectively (χ2=17.79,P=0. 000). By multivariate analysis, smoking history was a statistically significant unfavorable factor for OS in EDSCLC patients (versus no-smoking, hazard ratio (HR)=1.462, χ2=4.40, P=0.036). In addition, ≥4 cycles of chemotherapy and TRT were favorable prognostic factors ( ≥4 cycles vs ＜4 cycles, HR =0. 420,χ2 = 17. 17, P = 0. 000; ChT/TRT vs ChT, HR = 0. 634, χ2 = 6. 20, P = 0. 013). Conclusions Smoking is a independent unfavorable prognostic factor and ≥ 4 cycles of chemotherapy And TRT are independent favorable prognostic factors for OS in EDSCLC.

Objective To prenatally diagnose HA fetus with different clinical backgrounds. Methods Genetic tests were performed on 15 gravidas subjected for prenatal diagnosis of HA and different methods were employed for diagnosis according to the gestational weeks and clinical data. Amniotic fluid were taken from pregnant women within 23 gestational weeks for direct genotyping and indirect linkage analysis, since these women had probands with clear-cut mutations. Cordocentesis was performed for linkage analysis in pregnant women over 23 gestational weeks with probands whose types of mutation were unknown, while the FⅧ activity tests were carried out simultaneously. For the pregnant women over 23 gestational weeks without proband, cordocentesis was operated for measurement of FⅧ activity and karyotyping, but carriers of hemophilia A could not be detected in these cases. The introns 22 and 1 inversion of F8 gene were identified by long distance-polymerase chain reaction. Nucleotide sequencing was employed if the gene inversion could not be found and linkage analysis of 7 polymorphic markers, including DXS1108, F8Civs13, INTRON22,DXS1073,DXS9901, DXS15, DXS8069 and sex site (Amelo) were applied eventually. Identification of maternal blood contamination must be done before the tests. Results Fifteen samples were identified without maternal blood contamination. Five fetuses were diagnosed with hemophilia A. Meanwhile there were three pregnant women whose cord blood FⅧ activities were less than 1%. One of them was accompanied by trisomy 21; another had inversion mutation in introns 22 of F8 gene; the remaining one was identified with missense mutation in exon 23 (p. Arg2182Cys) of F8 gene. Conclusions Diverse methods should be applied in prenatal diagnosis of hemophilia A with different clinical backgrounds, for the sake of birth defects prevention.

Adult ; Female ; Humans ; Kidney Glomerulus ; metabolism ; pathology ; Lipoproteins ; metabolism ; Male ; Nephrosis, Lipoid ; metabolism ; pathology

OBJECTIVETo construct a eukaryotic expression vector for expressing hepatitis C virus (HCV) recombinant NS5B-EGFP fusion protein and obtain a stable transfected HepG2 cell line.

METHODSThe coding region of NS5B gene of HCV was amplified by PCR and was digested by Xho I/Kpn I. This fragment was inserted into pEGFPN3 with T4 ligase and transformed E. coli TG1. The positive recombinant plasmid was selected, then the recombinant plasmid was transfected into HepG2 cell by Lipofectin AMINE 2000. Cells containing stable transformants were selected by the ability of resistance to G418 and isolated with a limited dilution. The stable transfected cell line expressing high level NS5B-EGFP fusion protein was obtained.

RESULTSThe eukaryotic expression vector named pEGFPN3-ns5b was successfully constructed and the stable transfected HepG2 cell line expressing NS5B-EGFP fusion protein was obtained.

CONCLUSIONThe stable transfected HepG2 cell line could express NS5B-EGFP fusion protein, could be used for anti-HCV infection with ns5b gene as the target.

Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Viral Nonstructural Proteins ; genetics ; metabolism

OBJECTIVETo observe the curative effects of bone marrow stem cell (BMSC) and peripheral blood stem cell (PBSC) transplantations on the avascular necrosis of femoral head (ANFH).

METHODSTotally 122 ANFH patients (211 coxae) treated by BMSC or PBSC transplantations were enrolled from July 2004 to December 2006. All of them were classed to different stages according to the ARCO. Control group were desired as themselves before and after treatment. The puncture of femoral artery was conducted with digital subtraction angiography (DSA), and the tubes were inserted into medial femoral circumflex artery, lateral femoral circumflex artery and obturator artery with the cell suspensions were gradually poured into the arteries.

RESULTSThe joint pain, joint functions and walking distance of 122 patients were detected for the follow-up. Compared with before treatment, the calibers thickened; vessels increased and blood velocity quickened of femoral head blood-supply artery were observed in 15 patients after 6 months checked by DSA. The reduced areas of femoral head necrosis in 8 patients indicated the new bone formation between 12 and 24 months.

CONCLUSIONSAutologous BMSC and PBSC transplantation results in the new bone formation and improvement of ischemia in areas of femoral head necrosis at 6 months. The change of angiography was observed about 12 to 24 months after cell transplantation. The stem cell transplantation is convenient, safe and effective in the treatment of the ANFH with no adverse reaction, and can be considered as a new therapy of ANFH.

Bone Marrow Transplantation ; Femur Head ; blood supply ; pathology ; surgery ; Femur Head Necrosis ; surgery ; Follow-Up Studies ; Humans ; Ischemia ; surgery ; Peripheral Blood Stem Cell Transplantation ; Stem Cell Transplantation ; methods ; Transplantation, Autologous ; Treatment Outcome

This study is to investigate effects of genistein on rat femoral bone metabolic in vitro. Rat femoral tissues was isolated and randomly divided into two groups including control group and genistein (1 x 10(-5) mol x(-1)) group. Determinations of alkaline phosphatase (ALP) activity, calcium content and osteoprotegerin (OPG), type I-collagen (Collagen-I), RANKL, Runx-2 and bone morphogenetic protein (BMP-2) mRNA expression were done by real-time PCR. The results showed that 1 x 10(-5) mol x L(-1) genistein could increase the activity of ALP and contents of Ca, regulate bone metabolism activity of OPG, RANKL, BMP-2, Collagen-I and Runx-2 mRNA expression level. Genistein can significantly modulate bone metabolism related gene expression level of rat femoral tissue in vitro, and can increase calcium content and the activity of ALP.
Alkaline Phosphatase ; metabolism ; Animals ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Calcium ; metabolism ; Collagen Type I ; genetics ; metabolism ; Core Binding Factor Alpha 1 Subunit ; genetics ; metabolism ; Enzyme Activation ; drug effects ; Femur ; metabolism ; Gene Expression Regulation ; Genistein ; pharmacology ; Osteoprotegerin ; genetics ; metabolism ; Phytoestrogens ; pharmacology ; RANK Ligand ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction