The aerolysin genes (aerA) of BZ and NK isolates were cloned and sequenced. The sequence analysis showed that the partial aerA of BZ and NK isolates consisted of 1393 bp, encoding a protein of 464 amino acids. The similarity of nucleotide and amino acid sequences of aerA between BZ and NK isolates was 97.6% and 98.3% respectively. The nucleotide sequence of aerA of BZ strain exhibited 71.6% to 97.5% homology with other Aeromonas isolates, and the amino acid sequence exhibited 68.0% to 98.9% homology. The phylogenetic tree based on aerA nucleotide sequences from Aeromonas isolates was constructed with neighbor-joining method. It showed that there were three branches of aerolysin genes, and a close relation- ship among Aeromonas hydrophila isolates which were clustered into the same branch.

Objective To investigate the effect of advanced oxidation protein products (AOPPs) on human renal tubular epithelial cells(HK-2) autophagy.Methods HK-2 cells were stimulated with AOPPs.RT-qPCR and Western blot were used to determine the expression of autophagy related protein LC3-Ⅱ/LC3-Ⅰ,Beclin1 and p62;Western blot was utilized to examine the activation of p38 MAPK pathway.Then p38 MAPK inhibitor (SB203580) was added and co-processed with AOPPs.The change of autophagy was observed Also,autophagy inducer rapamycin was added and co-processed with AOPPs.RT-qPCR and Western blot were used to detect the expression of cell cycle inhibitory protein p27.The cell total protein level was detected by the bicinchoninic acid (BCA) method.The hypertrophy change was observed.Results AOPPs down-regulated the expression of LC3-Ⅱ/LC3-Ⅰ and Beclin1,up-regulated expression of p62 and activated p38 MAPK pathway;in comparison with the AOPPs alone treatment group,the expression of LC3-Ⅱ/LC3-Ⅰ and Beclin in the SB203580 co-processing group was increased,while p62 was decreased;the p27 expression and cells total protein in the sirolimus co-processing group were down-regulated.Conclusion AOPPs inhibits the autophagy of HK-2 cells by activating p38 MAPK pathway and the autophagy inhibition participates in HK-2 cell hypertrophy.

OBJECTIVETo discuss the effect of acupuncture for treatment of chronic functional constipation (CFC).

METHODSNinety cases were treated with acupuncture. The following two groups of acupoints were used alternatively once every other day. The acupoints in the first group were Tianshu (ST 25), Qihai (CV 6), Shangjuxu (ST 37) etc., and Zhongliao (BL 33), Xialiao (BL 34), Dachangshu (BL 25) etc. in the second group, electroacupuncture was used at Zhongliao (BL 33), Xialiao (BL 34), Tianshu (ST 25) and Shangjuxu (ST 37), once a day, 10 times constituting one course. The defecation frequency, difficulty degree of defecation, defecation time, endless sensation of defecation, stool quality and awareness of defecation were observed and the Patient Assessment of Constipation Quality of Life (PAC-QOL) was evaluated by constipation patients' diaries.

RESULTSThe scores of defecation frequency, difficulty degree of defecation, defecation time, endless sensation of defecation, stool quality, awareness of defecation and PAC-QOL were obviously improved after treatment (all P < 0.01). The total effective rate was 67.7% (61/90). The effect of acupuncture for chronic functional constipation in different dynamic mechanism was different. The effect of slow transit constipation (STC) was better than that of spastic pelvic floor syndrome (SPFS) (P < 0.05), and the effect of constipation caused by irritable bowel syndrome (IBS-C) was better than that of SPFS and relaxant pelvic floor syndrome (RPFS) (both P < 0.05). Fifty-two cases were effectively followed up. Three cases were cured, 6 cases were remarkably effective, 23 cases were effective and 20 cases were ineffective after 1 month of treatment. Three cases were cured, 5 cases were remarkably effective, 16 cases were effective and 28 cases were ineffective after 3 months.

CONCLUSIONThe effect of acupuncture for CFC with exact etiology, disease location and classification diagnosis is definite, but different dynamic mechanism has different effect. The treatment programs for SPFS and RPFS need to be optimized to improve the therapeutic effect.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Constipation ; physiopathology ; therapy ; Defecation ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the in vitro effect of HSV-tk/GCV using a hTERT promoter-driven vector system on Skov3 ovarian cancer cells.

METHODSAn expression vector (pBTdel-279-tk) containing tk gene under the hTERT promoter was constructed by molecular biological methods, and then was transfected into Skov3 ovarian cancer cells, normal ovarian epithelial cells (NOEC) and human embryonic lung fibroblast by cationic liposome. Following the transfection with tk, GCV was added, and MTT and flow cytometry methods were applied to investigate its antitumor effect. RT-PCR was used to detect the tk gene in ovarian cancer cells and normal cells after the transfection of pcDNA3-tk or pBTdel-279-tk.

RESULTSpBTdel-279-tk/GCV system induced apoptosis in hTERT-positive ovarian cancer cells, but not in hTERT-negative normal ovarian epithelial cells and fibroblasts. The hTERT promoter system was almost as efficient in inducing cancer cell death as the CMV promoter. tk gene was expressed in Skov3 cells and NOEC after pcDNA3-tk transfection, while positive was only in ovarian cancer cells after pBTdel-279-tk transfection.

CONCLUSIONThe telomerasespecific transfer of the tk gene under the hTERT promoter is a novel targeting approach for the treatment of ovarian cancer and may lead to an effective and specific gene therapy.

Apoptosis ; genetics ; Cystadenocarcinoma, Serous ; genetics ; pathology ; DNA-Binding Proteins ; Female ; Ganciclovir ; pharmacology ; Genetic Therapy ; Humans ; Ovarian Neoplasms ; genetics ; pathology ; Promoter Regions, Genetic ; genetics ; Simplexvirus ; genetics ; Telomerase ; genetics ; Thymidine Kinase ; genetics ; Transfection ; Tumor Cells, Cultured

OBJECTIVETo elucidate the reconstruction of neovascularization that occurred in the superiorly and inferiorly based posterior pharyngeal flaps in different time postoperatively.

METHODSTen mongrel dogs were randomly divided into two experimental groups, which were performed superiorly or inferiorly based posterior pharyngeal flap surgery respectively. Each group was then subdivided into five subgroups, and were sacrificed immediately after operation or on 3, 7, 30, 90 day postoperative respectively. Microangiography was used to exhibite the vessel.

RESULTS1. The blood vessel reconstruction of the superiorly based posterior pharyngeal flap was more rapid compared with the inferiorly based flap. The 3-day flap has established an axial vascular network, which was mature on the 30-day flap. The superiorly based posterior pharyngeal flap was mainly supplied by the pedicle. 2. The blood vessels reconstruction of the inferiorly based posterior pharyngeal flap was firstly occurred in the pedicle and apex of the flap, which grew slowly to the middle of the flap. The inferiorly based posterior pharyngeal flap was supplied by the pedicle and the soft palate. A mature axial vascular network was exhibited on the 90-day flap, which was not mature on the 30-day flap.

CONCLUSIONBoth superiorly and inferiorly based posterior pharyngeal flap can establish an axial vascular network and gain ample blood supply.

Animals ; Dogs ; Neovascularization, Physiologic ; Palate, Soft ; blood supply ; Pharynx ; blood supply ; Random Allocation ; Reconstructive Surgical Procedures ; Surgical Flaps ; blood supply

OBJECTIVETo investigate the effect of lithium chloride (LiCl) on cell cycle of HK-2 cells and explore the possible pathways involved.

METHODSHK-2 cells were treated with LiCl at different concentrations (5, 12.5, 20, and 25 mmol/L) for 12, 24, 48, or 72 h, and the changes in cell cycle and viability were detected using flow cytometry and CCK-8 assay, respectively. Western blotting was used to analyze the changes in the expressions of cyclin B1 and CDK1 (the two G2 phase-related proteins) and those of AKT/GSK-3β signaling pathway-related proteins in the treated cells.

RESULTSLiCl treatment time- and concentration-dependently increased HK-2 cell percentage in G2 phase and decreased the cell vitality. The expressions of cyclin B1, CDK1, p-GSK-3β, and β-catenin increased and the expression of p-AKT decreased significantly in the cells as LiCl treatment time and concentration increased.

CONCLUSIONLiCl may cause HK-2 cell cycle arrest in G2 phase through activation of the AKT/GSK-3β signaling pathway.

Objective The activation of NF-kappa B (NF-κB) signaling pathway plays an important role in the development of helicobacter pylori associated gastritis (HAG). The article aimed to investigate the effects of polygonum capitatum on the treatment of HAG in NF-κB signaling pathway and observe whether the regulation of NF-κB acetylation by silent information regulator 1 (SIRT1) affects the therapeutic effects of HAG. Methods The immortalized human gastric epithelial cells (GES-1) were cultured and the H.pylori stand- ard strain ATCC700392 was used for the replication of HAG cell model by 100∶1. The cells were divided into model group,drug group and normal control group. Cells were treated with 80 μg/mL in drug group,H.pylori and GES-1 were cultured together in model group and untreated GES-1 cells were taken as control group. Real time PCR was used to detect the mRNA levels of SIRT1,NF-κB/p65 and TNF-α. Western blotting was used to detect the expression levels of SIRT1,NF-κB/p65 and its acetylated protein in the total protein,as well as the expression levels of SIRT1 and NF-κB/p65 in cytoplasm and nuclear protein. Results At 12 h after the infection of H. pylori,the level of TNF-α in the supernatant was higher than that in the normal control group(P<0.05). The expression of SIRT1 de-creased in the cytoplasm of model group,while the expression levels of NF-κB/p65,acetyl-NF-κB p65(Lys310) and TNF-α in the nu-cleus increased (P<0.05). But after the treatment of polygonum capitatum,the expression of SIRT1 in the nucleus increased(P<0.05) while the expression of NF-κB/p65,acetyl-NF-κB p65(Lys310) and TNF-α decreased (P<0.05). Conclusion Polygonum capita-tum can activate the SIRT1 in the nucleus,which makes activated NF-κB/p65 in the nucleus carry out deacetylation modification in or-der to antagonize the cell damage induced by H.pylori.

OBJECTIVETo study the clinical, pathologic and radiologic features of chondroblastoma occurring in sites other than epiphysis and apophysis of long bones, and to investigate possible reasons for misdiagnosis.

METHODSThe clinical, pathologic and radiologic data of 18 chondroblastoma cases occurring in atypical sites were collected from 5 major hospitals in Shanghai during the past 12 years. S-100 immunostaining was performed to confirm the cartilaginous differentiation of the tumor cells.

RESULTSChondroblastoma occurred in small bones of feet in 10 of the 18 cases (55.6%) studied, being commonest in the talus and calcaneus bones. Mean age of the patients was 27.8 years, with 55.6% over 25 years of age. Radiologic examination revealed expansive, multilocular and well-demarcated radiolucent lesions in most cases. There was local cortical destruction in 5 cases (28%) and soft tissue infiltration in 1 case. In 10 cases (55.6%), the tumor was associated with aneurismal bone cyst or simple bone cyst formation. None of the cases studied was accurately diagnosed clinically before the operation. In 2 cases, the pathology was also misdiagnosed, often being diagnosed as aneurismal bone cyst or giant cell tumor.

CONCLUSIONSChondroblastoma occurring in atypical sites are often associated with atypical age, radiologic features and pathologic findings at presentation. Thorough understanding of the potential pitfalls is essential in order to avoid misdiagnosis.

Adolescent ; Adult ; Bone Cysts, Aneurysmal ; diagnosis ; diagnostic imaging ; pathology ; Bone Diseases ; diagnosis ; diagnostic imaging ; pathology ; Bone Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; Calcaneus ; Child ; Chondroblastoma ; diagnosis ; diagnostic imaging ; pathology ; Diagnostic Errors ; Female ; Giant Cell Tumor of Bone ; diagnostic imaging ; pathology ; Humans ; Male ; Radiography ; Talus

OBJECTIVETo investigate whether the p38 mitogen-activated protein kinase (MAPK) signaling pathway mediates advanced oxidation protein products (AOPPs)-induced epithelial-to-mesenchymal transition (EMT) in tubular cells.

METHODSHuman proximal tubular cells (HK-2 cells) exposed to AOPP-bovine serum albumin (BSA) were examined for expressions of p38 MAPK and phosphorylated p38 MAPK using Western blotting. Western blotting and quantitative RT-PCR were used to examine the protein and mRNA expressions of EMT markers E-cadherin and vimentin and endoplasmic reticulum stress marker glucose-regulated protein (GRP) 78 in cells treated with SB203580 (an inhibitor of the p38 MAPK signaling pathway) prior to AOPP exposure. The cells treated with AOPPs following pretreatment with salubrinal (an inhibitor of endoplasmic reticulum stress) were also examined for expressions of p38 MAPK and phosphorylated p38 MAPK.

RESULTSAOPP treatment induced the phosphorylation of p38 MAPK in HK-2 cells. AOPP-induced decrease in E-cadherin expression and overexpression of vimentin and GRP78 were partly inhibited by pretreatment of the cells with SB203580. Salubrina partly suppressed AOPP-induced phosphorylation of p38 MAPK in the cells.

CONCLUSIONp38 MAPK signaling pathway, which is regulated by endoplasmic reticulum stress, might mediate AOPP-induced EMT in HK-2 cells.

OBJECTIVETo establish baseline patterns of rotavirus diarrhea and to describe its epidemiologic features in Changchun city, prior to rotavirus vaccine immunization.

METHODSHospital-based surveillance was conducted among children under 5 years old with acute diarrhea in Changchun Children's Hospital. Fecal samples were determined to identify rotavirus by PAGE and/or ELISA. G serotypes of rotavirus were identified by ELISA and/or nested RT-PCR. P genotyping were carried out by RT-PCR. All data were computerized and analysed by "Generic Manual on Rotavirus Surveillance" set by CDC in the USA.

RESULTSIn total, 2 343 diarrhea cases were screened and 1 211 fecal samples were collected. Rotavirus was detected in 31.0% among outpatients and 52.9% in inpatients. During the peak of the season (November through March), 58.6% of diarrhea was caused by rotavirus among inpatients. 95.0% of rotavirus diarrhea cases occurred among children aged < 2 years. The predominant strain was serotype G1 (82.4%), followed by G2 (5.0%), G3 (3.3%), G4 (0.9%). P genotyping showed that P[8] and P[4] were the most common ones. Nine different P-G combinations were identified, four strains (P[8]G1, P[4]G2, P[8]G3, and P[8]G4) commonly seen worldwide accounted for 75.6% of the total. Taken together with uncommon strains, including the novel types P[4]G4 and P[8]G2, it highlights the extraordinary diversity of rotaviruses circulating in China.

CONCLUSIONRotavirus is the major cause of severe child diarrhea in Changchun. Developing a rotavirus vaccine for prevention of severe disease and reduction of treatment costs seemed to be necessary.

Child, Preschool ; China ; epidemiology ; Diarrhea ; etiology ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Feces ; virology ; Female ; Genotype ; Hospitals, Pediatric ; statistics & numerical data ; Humans ; Infant ; Male ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; complications ; epidemiology ; virology ; Sentinel Surveillance ; Serotyping