A novel nucleic acid amplification method,termed loop-mediated isothermal amplification(LAMP),which amplifies DNA with high specificity,efficiency,and rapidity under isothermal conditions,may be a valuable tool for the rapid detection of infectious diseases.This method employs a DNA polymerase that have activity of strand displacement DNA synthesis and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA.LAMP can amplify a few copies of DNA to 109 in less than an hour.The final products are stem-loop DNA with several inverted repeats of the target and cauliflower-like structures with multiple loops.A positive reaction would be shown as a ladder-like pattern in a gel electrophoresis analysis.Because of the advantage,the LAMP method will be widely applied to research of nucleic acid,clinical diagnosis of infectious diseases and detection of genetically modified organisms etc.

Objective To explore the effects of modified tubo-uterine implantations performed on women with proximal tubal occlusive infertility after femal sterilization with mucilago phenol.Methods Two hundred and eight infertile women who were admitted to the Second Affiliated Hospital of Sun Yat-sen University between 1986 and 2004 were included.They all accepted modified tubo-uterine implantation after occlusion of fallopian tubes with mucilago phenol.Results It was found that the occlusions were all located in the interstitial portion or isthmic portion of the fallopian tubes.Different degrees of pelvic adhesions were found in 65 cases.Fifty-seven cases were slightly adhesive,seven cases were of moderate degree and one case was severe.One hundred and ninety-nine cases were followed up after operations(95.7%).One hundred and ninety-three women accepted hydrotubation in the following month just after the operation and 185 women were found to be unobstructed(95.8%).One hundred and forty-three women became pregnant, the pregnant rate being 71.9%(143/199).One hundred and twenty-five women had term deliveries (87.4%),three women were in early pregnancy and two in midtrimester pregnancy.Eleven women had spontaneous abortion(7.7%).Two women had tubal pregnancy(1.0%).None of the 199 cases had any signs of endometriosis.Conelusions Modified tubo-uterine implantations are quite effective for proximal tubal occlusive infertility.It may be a favorable method for such kind of tubal occlusions.

Objective To study mental characteristics of children with light lead poisoning. Methods Eighty - six children with light lead poisioning and 33 control children were evaluated with WISC - R and social adaptive capacity(SAB). Results 1. Intelligence quotient( IQ)and SAB of children with light lead poisoning were lower than normal children. 2. There was also deficiency of perception factor and memory/ concentration factor among children with lead poisioning and the blood lead level had negative association with perception factor. Conclusion Light lead poisioning may affect children's IQ, especially in the perception factor and memory /concentration factor.

Adult ; Aged ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Mucin-2 ; Mucin-6 ; Mucins ; analysis ; Neoplasms, Cystic, Mucinous, and Serous ; chemistry ; Pancreatic Neoplasms ; chemistry

OBJECTIVETo investigate the effects and related mechanisms of hepatitis B virus X (HBx) protein on cell cycle and growth in hepatocellular carcinoma.

METHODSA human hepatocyte HepG2 cell line stably expressing a green fluorescent protein (GFP)-tagged HBx (HepG2/GFP-HBx cells) was used for the experiment, and HepG2 parental and HepG2/GFP cells was used as the controls. Effect of HBx on cell growth was evaluated by the MTT cell proliferation assay and on cell cycle progression by flow cytometry analysis of cells with or without treatment with 5-aza-2'-deoxycytidine (5-Aza-CdR; 5 pmol/L). Effect of HBx expression on promoter methylation status of the p16INK4A tumor-suppressor gene was detected by methylation-specific polymerase chain reaction and on p16 protein level was analyzed with western blotting.

RESULTSThe HepG2/GFP-HBx cells showed significantly higher cell proliferation at 72 hrs of culture (3.225+/-0.038 A490) than either control (HepG2: 2.012+/-0.022 A490, t = -46.86, P less than 0.001; HepG2/GFP: 2.038+/-0.029 A490, t = 42.51, P less than 0.001). The HepG2/GFP-HBx cells also showed significantly lower proportion of cells in the G0/G1 phase (16.45%+/-0.45%) than either control (HepG2: 44.81%+/-1.36%, t = -34.202, P less than 0.001; HepG2/GFP: 42.76%+/-1.58%, t = -28.88, P less than 0.001). However, 5-Aza-CdR treatment did lead to a significant amount of HepG2/GFP-HBx cells being arrested in the G0/G1 phase (33.25%+/-0.79%, t = 31.85, P less than 0.001). The p16INK4A promoter was methylated in the HepG2/GFP-HBx cells, and became demethylation after treatment with 5-Aza-CdR. However, no methylation of p16INK4A promoter was observed in both HepG2 and HepG2/GFP cells. The p16 protein level was significantly lower in the HepG2/GFP-HBx (vs. HepG2 and HepG2/GFP cells) and this level increased after treatment with 5-Aza-CdR.

CONCLUSIONHBx protein promotes hepatocellular carcinoma cell cycle progression and growth by shortening the G0/G1 phase, and the underlying mechanism may involve inducing p16INK4A promoter methylation and downregulating p16 protein expression.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Genes, p16 ; Hep G2 Cells ; Hepatitis B virus ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Promoter Regions, Genetic ; Trans-Activators ; pharmacology

Objective To analyze the medical environmental distribution characteristics of carbapenem-resistant Klebsiella pneumoniae (CRKP).Methods On April 25 and August 29,2016,random sampling was performed be-fore cleaning and disinfecting the surrounding of patients who isolated CRKP in a hospital,sampling were mainly object surfaces and medical devices.Distribution characteristics of CRKP were analyzed.Results The number of environmental sampling in surgical intensive care unit(SICU),respiratory ICU(RICU),and emergency ICU(EICU) were 90,41,and 56 respectively.CRKP was not found in SICU and RICU specimens;6 strains of CRKP were iso-lated in EICU,the isolation rate was 10.71%.In April 2016,103 specimens were taken,and 6 strains of CRKP were isolated,isolation rate was 5.83%,which was higher than in August of 2016(P <0.05).There were no sig-nificant differences in isolation rates of CRKP among different specimens and different responsible persons (all P >0.05).Conclusion Key departments,such as EICU,should be intensified cleaning and disinfection,so as to re-duce the spread of CRKP in the medical environment.

OBJECTIVETo observe the therapeutic effect of eye-needling combined with medication for treatment of ophthalmoplegia and explore the possible mechanism.

METHODSOne hundred and twenty cases were randomly divided into a treatment group and a control group. According to etiological factors, the control group were treated with medication and the treatment group with the medication plus eye-acupuncture at main point ocular muscles. Changes of the rima oculi, the range of ocular movement and the dialopia angle after treatment were recorded and statistically analyzed in the two groups.

RESULTSThe total effective rate was 93.4% and the cured rate was 54.1% in the treatment group, and 74.6% and 18.6% in the control group, with significant difference between the two groups (P < 0. 01).

CONCLUSIONEye-needling combined with medication has an obvious therapeutic effect which is better than simple medication for ophthalmoplegia.

Acupuncture Therapy ; methods ; Adolescent ; Adult ; Aged ; Child ; Combined Modality Therapy ; Female ; Humans ; Male ; Middle Aged ; Ophthalmoplegia ; therapy

OBJECTIVETo explore the mucosal protective effect on the quality of gastric ulcer healing.

METHODSGastric ulcers were induced in male rats by serosal application of acetic acid. Rats were gavaged for 14 days with saline, omeprazole (OME), teprenone (TEP) and TEP plus OME starting 3 days after ulcer induction. Then the tissues and blood samples were obtained and measured.

RESULTThe lower ulcer index (UI) and increased ulcer inhibition rate were observed in OME and OME+TEP groups. In TEP and OME+TEP groups, restored mucosa thickness increased, cystically dilated glands decreased, microvessels in connective tissue increased, the secretion of mucus, hexosamine, PGE(2), bFGF were enhanced, the expression of EGFR was increased.

CONCLUSIONTEP can improve the quality of gastric ulcer healing, when combined with OME,the effect is more marked.

Acetic Acid ; Animals ; Anti-Ulcer Agents ; therapeutic use ; Diterpenes ; therapeutic use ; Drug Therapy, Combination ; Gastric Mucosa ; drug effects ; metabolism ; pathology ; Male ; Omeprazole ; therapeutic use ; Rats ; Rats, Wistar ; Receptor, Epidermal Growth Factor ; biosynthesis ; Secondary Prevention ; Stomach Ulcer ; chemically induced ; drug therapy ; pathology ; Wound Healing ; drug effects

Purpose To investigate the expression and the methylation status of miR-4687-5P and STIM1 gene in esophageal squamous cell cancer (ESCC) cell lines and ESCC tissue samples,in order to explore the correlation between miR-4687-5P and STIM1 expression,as well as whether they have a common expression regulation mechanism. Methods The qRTPCR and methylation specific PCR (MSP) methods were applied respectively to examine the expression and methylation of miR-4687-5P and STIM1 genes in ESCC cell lines (TE13, KYSE150,T. Tn) and ESCC samples,and further to analyze their correlation. Results The expression of miR-4687-5P and STIM1 genes in ESCC was significantly decreased,and consistent. The weak expression of miR-4687-5P and STIM1 genes was detected in three ESCC cell lines. After treated with 5-Aza-2'-deoxycytidine (5-Aza-Dc,a demethylation agent) ,the expression levels of these two genes were obviously increased. Meanwhile, the methylation bands were obviously weakened or disappeared. The promoter region of STIM1 gene was hypermethylated in ESCC tissues,and its methylation frequency was correlated with the expression of STIM1 and miR-4687-5P (P < 0. 01) . Conclusion miR-4687-5P and STIM1 genes are down-regulated in esophageal carcinoma,and the expression of miR-4687-5P may be regulated by the promoter of its host gene STIM1,and the hypermethylation may be one of the common mechanisms leading to down-regulatory expression of miR-4687-5P and STIM1 genes in ESCC.

Brain Injuries ; pathology ; surgery ; China ; Craniotomy ; Earthquakes ; Hematoma, Epidural, Cranial ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Time Factors ; Treatment Outcome