Objective To investigate the role and intracellular signal transduction mechanism in the injury of renal tubular cells induced by postasphyxial-serum in neonate.Methods Human renal proximal tubular cell(HK-2 cell) was used as target cell. The experiment was designed as:control group, asphyxia group ,and pyrrolidine dithiocarbamate (PDTC)blocking group. The attacking concentration of serum was 20%, and the apoptosis rate of HK-2 cells was detected by flow cytometer.Results Compared with controls[(13.3?1.70)%],after being stimulated with postasphyxial-serum, the apoptosis rate of HK-2 cells of asphyxia group [(46.73?3.68)%] and PDTC blocking group [(31.19?2.79)%]were significantly increased(P

Objective To investigate the role of postasphyxial-serum of neonate in inducing injury of human renal proximal tubular cells(HK-2 cells).Methods HK-2 cells were used as target cell.The neonatal different concentration postasphyxial-serum of 1,3,7 days after asphyxia were used as attacking means.The experimental groups were divided into 15 groups:the 2.5%,5.0%,10.0%,(20.0%) attacking concertration groups of 1,3,7 day after asphyxia and control group of each concertration.The culture medium and concertration of the control group and the experimental groups were the same.The changes of morphology were observed under inverted microscope,the cell viability was measured by 3-(4,5-dimethyl-2-thiazoly1)-2,5-diphenyl-2H-tetrazolium bromide(MTT) method and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,the changes in morphology of HK-2 were most serious and obvious,the cell viability were obviously decreased(all P

Abstract: Objective　To investigate the distribution and drug resistance of pathogenic bacteria separated from ascites of patients in Children’s Hospital Affiliated to Zhengzhou University from 2015 to 2021, and to provide a basis for rational clinical antimicrobial agents. Methods　Bacterial culture, bacterial identification and drug sensitivity analysis were performed on 1 058 non-duplicate ascites culture specimens from January 2015 to December 2021. The clinica1 and microbiologica1 data were ana1yzed by WHONET 5.6 and SAS 9.4 Results　Of the 1 058 specimens, 586 (55.39%) were positive for pathogenic bacteria, with a total of 781 strains isolated. There was no significant trend of increase or decrease in the positivity rate over different years. Male children (63.99%) were more prevalent than female children. Appendicitis (59.22%) was the most common disease and Escherichia coli was the most common causative bacteria. Among neonates (≤28 d), the bacteria with the highest detection rate were Klebsiella pneumoniae (23.50%) and Enterococcus faecium (23.50%), while among children (>28 d), the highest detection rate was Escherichia coli (35.98%). Gram-negative bacteria accounted for 64.79% of the 781 strains, mainly Escherichia coli (38.28%), Klebsiella pneumoniae (8.58%), and Pseudomonas aeruginosa (5.89%); Gram-positive bacteria accounted for 29.45%, mainly Enterococcus faecium (8.58%), Streptococcus constellatus (2.69%), and Enterococcus avium (2.43%); fungi accounted for 1.66% and anaerobic bacteria accounted for 4.10%. The resistance rates of Escherichia coli to cefoperazone/sulbactam, piperacillin/tazobactam, imipenem and meropenem were 6.02%, 4.35%, 4.35%, and 3.68%, respectively. The resistance rates of Klebsiella pneumoniae to these drugs were 59.70%, 59.70%, 50.75% and 53.73% respectively. Linezolid-resistant strains of Enterococcus faecium were found. Conclusion　Appendicitis is the most common abdominal infection in children, and the distribution of ascites pathogens varies with ages and diseases. The pathogenic bacteria are mainly Gram-negative bacteria, and the drug resistance of Klebsiella pneumoniae was more serious. It is particularly important to use antibiotics correctly and rationally to reduce the emergence of drug resistant bacteria.

Anti-Bacterial Agents ; therapeutic use ; Child, Preschool ; Female ; Femoral Artery ; diagnostic imaging ; surgery ; Humans ; Lower Extremity ; blood supply ; Lung ; diagnostic imaging ; pathology ; Male ; Mycoplasma pneumoniae ; Pneumonia, Mycoplasma ; complications ; microbiology ; therapy ; Thrombolytic Therapy ; methods ; Thrombosis ; etiology ; microbiology ; therapy ; Tomography, X-Ray Computed

OBJECTIVETo explore a new method for early avascular necrosis of femoral head (AVNFH) therapy.

METHODSSixty-nine AVNFH New Zealand adult rabbits were randomly divided into three groups with equal number. In Group A, deproteinized bone (DPB) that absorbed with recombinant plasmid pcDNA3.1-hVEGF165 was implanted into the drilled tunnel of necrotic femoral head. In Group B, only DPB was implanted. In Group C, only tunnel was drilled without DPB or plasmid implanted. Femoral head specimens were obtained at postoperative 1, 2, 4, 8, 16 weeks. The expression of VEGF165 and collagen I was detected by immunohistochemistry. Bone formation was detected generally by X-ray. Angiogenesis and the repair of the femoral head were observed histologically.

RESULTSThe expression of VEGF 165 could be detected 2 weeks after implantation in Group A, but it was not observed in other groups. The result of collagen I expression had a significantly difference 2, 4 and 8 weeks after operation in Group A from those in other groups (P < 0.01). X-ray results indicated that there was more bone formation in Group A than in other groups. The regenerated capillary vessels staining result of necrotic femoral head in Group A was significantly different from those in other groups at postoperative 2 and 4 weeks (P < 0.01).

CONCLUSIONSTransfection of hVEGF165 gene enhances local angiogenesis and DPB-VEGF compound improves the repair of necrotic femoral head. Deproteinized bone grafting with VEGF gene transfer provides a potential method for the treatment of osteonecrosis.

Animals ; Bone Transplantation ; Collagen Type I ; analysis ; Femur Head Necrosis ; pathology ; physiopathology ; therapy ; Genetic Therapy ; Immunohistochemistry ; Neovascularization, Physiologic ; Rabbits ; Transfection ; Vascular Endothelial Growth Factor A ; analysis ; genetics

Five structural important residues of rice nonspecific lipid transfer protein LTP110 were mutated by site-directed mutagenesis. Sequence results showed that they were all mutated successfully. After trying various E. coli expression systems, thioredoxin fusion expression system was found to be a proper system to express wild type and mutant LTP110. cDNA sequences encoding wild type LTP110 and the mutants Y17A, P72L, R46A, D43A, C50A were cloned into two kinds of thioredoxin fusion expression vectors. The expression results were compared. In pTrxFus/GI724 expression system, wild type LTP110 and the mutants Y17A, P72L, R46A could be expressed at low level while D43A and C50A could not be expressed normally; in pET32a(+)/BL21 (DE3) trxB- expression system, wild type LTP110 and all mutant proteins could be expressed very well and the levels were higher than that in pTrxFus/GI724 system. LTP110 fusion protein expressed in pET32a(+) vector was purified and its activity was checked by fluorescence labeled fatty acid. Results indicated that the recombinant LTP110 fusion protein has lipid binding activity. This work provides good basis for the further study.
Amino Acid Sequence ; Carrier Proteins ; genetics ; isolation & purification ; metabolism ; Gene Expression ; Genetic Engineering ; Genetic Vectors ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Oryza ; genetics ; Plant Proteins ; genetics ; isolation & purification ; metabolism ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; metabolism ; Thioredoxins ; genetics

Objective To study the feasibility and clinical values of diffusion tensor tractography (DTT)in the spinal cord at 3.0 T MR.Methods Forty patients with spinal cord compression including cervical cord herniation and cervical spondylosis(30 cases),tumors in spinal canal(9 cases)and old injury in cervical vertebrae(1 cases)and 20 healthy volunteers participated in this study.Single-shot spin- echo echo-planar diffusion tensor sequence for tractography of the spinal cord was performed.The fibers of spinal cord were visualized by using fiber tracking software.Results On the DTT maps,the normal spinal cord was depicted as a fiber tract showing color-encoded cephaloeaudally,which indicated anisotropy in the cephalocaudal direction.By setting two ROI,the main spinal cord fiber tracts,such as corticospinal or spinothalamic tract,were visualized.The tracts from two sides of the brain did not completely cross.It was asymmetric in the number of tracts on the two sides in most normal subjects(8/10).The tracts of all patients with cord compression were seen oppressed or damaged in different degrees.The DrrT in patients with cervical spondylosis and extramedullary-intradural neurolemmoma demonstrated that tracts were oppressed but not damaged.The DTT in one ependymoma showed that tract was markedly compressed and slightly damaged.Conclusion DTT is a promising tool for demonstrating the spinal cord tracts and abnormalities,can provide useful information for the localization of compression and evaluation of the impairment extent on the white matter tracts of the spinal cord.

Adult ; Carcinoma, Hepatocellular ; Female ; Humans ; Liver Neoplasms ; Pregnancy ; Pregnancy Complications, Neoplastic

OBJECTIVETo evaluate the efficacy and safety of intravesical instillation with gemcitabine after first-line intravesical chemotherapy failure, including mitomycin (MMC), epirubicin (EPB) and camptothecin (CPT), in the treatment of non-muscle-invasive bladder cancer (NMIBC).

METHODSFrom June 2007 to October 2008, 72 patients with NMIBC, who had tumor recurrence within one year of first-line intravesical chemotherapy, were assigned to 3 groups (24 cases each). Group A received intravesical gemcitabine in a dose of 1000 mg, Group B received 2000 mg gemcitabine, and Group C received original intravesical chemotherapy. The time of reccurrence and adverse effects were recorded.

RESULTSThe 2-year tumor free survival rates of the 3 groups were 66.7%, 75.0% and 45.8%, respectively. The 2-year TFS rate of the patients who received gemcitabine was 70.8%, significantly higher than 45.8% of the patients treated by original chemotherapy. There was one case with renal function impairement in the groups A and B, respectively. There was no significant difference between the rates of low urinary tract symptoms in the 3 groups. No severe hematological side effects were observed in this study.

CONCLUSIONThe intravescal chemotherapy with gemcitabine in patients with recurrent bladder tumor after first-line intravesical chemotherapy is effective and well tolerated, however, renal function should be routinely assessed.

Administration, Intravesical ; Adult ; Aged ; Antibiotics, Antineoplastic ; therapeutic use ; Antimetabolites, Antineoplastic ; administration & dosage ; adverse effects ; therapeutic use ; Antineoplastic Agents, Phytogenic ; therapeutic use ; Camptothecin ; therapeutic use ; Deoxycytidine ; administration & dosage ; adverse effects ; analogs & derivatives ; therapeutic use ; Disease-Free Survival ; Dose-Response Relationship, Drug ; Epirubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Mitomycin ; therapeutic use ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; Urinary Bladder Neoplasms ; drug therapy ; pathology

OBJECTIVETo investigate the effects of angelicanaphtha on proliferation, cell cycle, apoptosis, and collagen synthesis of human umbilical vein endothelial cells (HUVEC).

METHODSHUVEC was cultured and passaged in Dulbecco's modified Eagle's medium (DMEM) and treated with angelicanaphtha 1 mg/ L, 4 mg/L, and 16 mg/L at 1, 3, 5, and 7 day respectively. The proliferation was measured with MTT method. The cell cycle and apoptosis were analyzed with flow cytometry and collagen synthesis was determined with radioimmunoassay.

RESULTSThe proliferation of the HUVEC was accelerated by angelicanaphtha < or =4 mg/L and inhibited by angelicanaphtha at 16 mg/L (P < 0.05). Lower concentration (< or = 4 mg/L) of Angelicanaphtha decreased cells in G0/G1 phase and increased significantly cells in S phase and inhibited the apoptosis (P < 0.05). However, angelicanaphtha at 16 mg/L increased cells in G0/G1 phase and decreased cells in S phase and induced the apoptosis (P < 0.05). The collagen synthesis of HUVEC was inhibited by angelicanaphtha in concentration-dependent manner (P < 0.05 or 0.01).

CONCLUSIONThe proliferation effects of angelicanaphtha on HUVEC had dualistic regulation of increase by lower-concentration and inhibition by higher-concentration. Collagen synthesis of HUVEC was inhibited by angelicanaphtha in concentration-dependent manner.

Angelica sinensis ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type III ; biosynthesis ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Oils, Volatile ; pharmacology ; Umbilical Veins ; cytology