The stool specimens of 235 patients with acute diarrhea were isolated and identifiedfor detection of ETEC,according to modified Elek test by purified anti-heat-labile entero-toxin serum and to the suckling mouse assay.Simultaneously,the other enteropathogenicorganisms were also isolated.33 strains of ETEC(14.04%)were obtained in total,amongwhich 29 strains(87.8%)belong to heat-stable enterotoxin(ST)type;3 strains(9.09%)heat-labile enterotoxin(LT)type and only 1 strain(3.1%)ST and LT mixed type.It wasshown that ETEC was one of the important enteric pathogens of patients with acute diar-rhea.The isolation,identification and incidence of ETEC were discussed briefly.

Objective To evaluate the clinical effect of lumbar intervertebral disc herniation (LIDH) by CT-guided precise injection around nerve root and in epidural cavity. Methods One hundred and eight patients of LIDH were treated by CT-guided precise injection around nerve root and in epidural cavity, and followed-up after 1, 3 and 6 months. Visual analogue score (VAS) for pain was used to evaluate the efficacy before and after treatment. Results In 108 cases, there were 83 patients (76.9%) with good result whose improvement of VAS was greater than or equal 50% after treatment 1, 3 and 6 months. There were 19 patients (17.6%) with fair result whose improvement of VAS was less than 50% after treatment 1, 3 and 6 months. There were 6 patients (5.6%) with invalid result whose improvement of VAS was unobvious after treatment 1, 3 and 6 months. Conclusion CT-guided therapy of LIDH by precise injection around nerve root and in epidural cavity is safe, accurate, effective, minimally aggressive technique and worth being practiced clinically.

Objective To study the clinical features of complicated upper urinary tract infection in children,aiming to provide a reference for clinical diagnosis and treatment of the disease.Methods The clinical data of 68 cases with complicated upper urinary tract infection hospitalized at the Department of Urinary Surgery NO.1,Children's Hospital of Shenzhen between January 2013 and August 2015 were retrospectively analyzed.Results A total of 68 cases,in which 44 were male,24 were female,and repeated infections were found in 15 cases.Fever was the most common clinical manifestation(48 cases,70.59％),while frequent urination and odynuria were rare(9 cases,13.24％).A total of 57 strains had been cultured form the submitted specimens which were obtained from the 68 cases,including 41 strains of gram-negative bacteria,15 strains of gram-positive bacteria and 1 strain of fungus.Two different strains were cultured in 3 children.Fourteen strains of Escherichia coli had 11 extended spectrum beta lactamases (ESBLs)-positive strains.In the 11 strains of klebsiella pneumoniae,8 trains were ESBLs-positive.The drug resistant rates of gram-negative bacteria to Ampicillin and Cefuroxime sodium were both more than 90％.The sensitive rates to Piperacillin/Tazobactam was more than 90％.Thirty-one cases of complicated upper urinary tract infection were cured by administering Piperacillin/Tazobactam,while 15 cases were cured by changing Cefuroxime sodium to Piperacillin/Tazobactam according to the drug sensitivity results.Conclusions Clinical manifestations of complicated upper urinary tract infection are untypical,and fever is the most common symptom.Repeated infection is common.The gram-negative bacteria is the dominant pathogen causing the complicated upper urinary tract infection.ESBLs-positive bacteria accounts for high proportion.The drug resistance rate to penicillin and the first,second generation of the cephalosporin is high.The drug sensitive rate of piperacillin tazobactam is high,with good prognosis.

Objective To investigate the clinical efficacy of 0.1％ mometasone furoate cream in the treatment of phimosis in children.Methods A prospective study was carried out over two years period on an outpatient which basis on two groups of patients with severe phimosis.598 children with severe phimosis (Kikiros classification 4-5) aged from 2 years old to 11 years old and 8 months were selected.311 cases in the observation group and 287 cases in the control group.The observation group applied a steroid cream the foreskin twice a day for 4 weeks,and the control group used local handling of the foreskin twice a day for 4 weeks.The effects of the two groups after 2 and 4 weeks of treatment were compared.Results 29 cases in the observation group and 47 cases in the control group were loss of follow up.In the steroids group which including 282 patients,68.8％ of patients (194 cases) showed a complete response (full retraction of the foreskin) to the therapy.The total efficiency rate of the 4 stage phimosis group is higher than the 5 grade phimosis group.Patients who had a history of balanoposthitis or urinary tract infection showed poorer improvement in preputial retraction.A total of 28 out of 240 patients (11.7％) in the control group showed a complete response to the therapy.The total efficiency rate of the observation group was significantly higher than the control group (x2 =173.121,P ＜ 0.01).There were 4 cases of discomfort in the observation group and 6 cases of foreskin injured in the control group.Conclusions Topical application of 0.1％ mometasone furoate cream in the treatment of severe phimosis in children is an effective,safe and simple non-invasive treatment with less adverse reactions.

Objective To explore the mechanism of apoptosis after cryotherapy on tumors.Methods (1) GL261 glioma cells (1×107 cell/10 μL) were injected into the subcutaneous one ofC57 mice to establish tumor-bearing mouse models;when the diameter of tumor reached to 15-20 mm,the mice were randomly divided into cryogenic treatment group and sham-operated group (n=1 0);mice in the cryogenic treatment group were given surgical cryotherapy,while those in the sham-operated group only performed surgery without cryotherapy.TUNEL was used to detect the cell apoptosis in glioma tissues 12 and 24 h after operation;and Western blotting was employed to detect the protein expressions of pro-caspase-8,pro-caspase-9 and poly-ADP-ribose polymerase (PARP).(2) GL261 glioma cells were divided into control group and one time cryogenic release group,and DMEM and one time of cryogenic release were given to the two groups,respectively;12 h after the treatment,TUNEL was used to observe the cell apoptosis in glioma tissues,and Western blotting was employed to detect the protein expressions.Results (1) TUNEL indicated that the cells in the S1 region of the glioma tissues from mice in the cryogenic treatment group were uniformly died;significant apoptosis was noted in cells of the S2 region at 12 h after treatment;while,24 h after treatment,S1 region still showed uniform necrosis,S2 region showed apoptotic regression,and S3 region showed new apoptosis at the target side.Western blotting indicated that pro-caspase-9 and PARP protein expressions at the S2 region were signficantly decreased as compared with those at the S1,S3 and S4 regions (P＜0.05),and pro-caspase-8 protein expression at the S3 region were signficantly reduced as compared with those at the S1,S2 and S4 regions in the cryogenic treatment group (P＜0.05).(2) TUNEL showed that significantly increased GL261 glioma cell apoptosis rate was noted in the one time cryogenic release group as compared with that in the control group (P＜0.05);Western blotting indicated that as compared with the control group,the cryogenic release group had significantly decreased pro-caspase-8 and pro-caspase-9 expressions (P＜0.05).Conclusion Cryogenic release or substances released from tumor tissues after cryoablation shows an effect on promoting apoptosis.

With the rapid development and adaptation of high-throughput sequencing in clinical settings, application of exome sequencing (ES) has been gradually expanded from pediatric to prenatal diagnosis in recent years. There is an urgent need to establish criteria for clinical grade ES in order to facilitate such a complex testing. The standardization of pre- and post-test consultation, quality control for sample processing process and validation of bioinformatics data analysis, and more importantly data interpretation and reporting, as well as appropriate reporting scope, is of great importance for health care stakeholders. To achieve this, a committee composed of a wide range of healthcare professionals has proposed an ES standard for prenatal diagnosis. This has provided expert opinion on the genetic counseling and reporting standards of prenatal ES for the purpose of applying ES technology in prenatal setting.

OBJECTIVE: To investigate the operative effect and value of window partial laryngectomy with endoscopy for treatment of stage T1b--T3 laryngeal cancer. METHOD: Twenty-seven cases with glottic laryngeal cancer were treated by window partial laryngectomy with endoscopy. After 3-year and 5-year survival rate and laryngeal functions (respiratory, pronunciation, swallowing function) recovery were evaluated. RESULT: The 3- and 5-year survival rates were 83.3% (15/18) and 75.0% (9/12) respectively. The decannulation rate was 92.6% (25/27) 4 weeks after operation. Three months after decannulation of respiratory function evaluation: the march to fast runner 77.8% (21/27), the jogger 14.8 (4/27), who walk briskly 7.4% (2/27). All patients had recovered after four weeks swallowing without choking cough and satisfied with phonation. Local recurrence rates at 3 and 5 year after operation were 10.5% (2/19) and 15.4% (2/13) respectively. CONCLUSION The research shows that window partial laryngectomy with endoscopy was applicable to stage T1b--T2 and selecting T3 laryngeal cancer patients. Postoperative respiratory, pronunciation and swallowing function recovery rate is satisfactory.
Adult ; Aged ; Carcinoma, Squamous Cell ; pathology ; surgery ; Female ; Follow-Up Studies ; Glottis ; pathology ; Humans ; Laryngeal Neoplasms ; pathology ; surgery ; Laryngectomy ; methods ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Survival Rate ; Thyroid Cartilage ; surgery ; Treatment Outcome

BACKGROUNDTo investigate the influence of the tumor metastasis suppressor gene nm23 H1 on the activity of extracellular signal-regulated protein kinase (ERK) in human high metastasis large cell lung cancer cell line L9981.

METHODSThe levels of total ERK1/2 and phospho-pERK1/2 were determined with p44/42 MAP kinase antibody and dually phosphospecific phospho-44/42 MAP kinase antibody in human high-metastasis large cell lung cancer cell lines L9981 (cell line with nm23-H1 gene deletion), L9981-nm23-H1 (cell line with nm23-H1 transfected ) and L9981-PLXSN (cell line with vector transfected) by Western blot method, respectively. The activity of phospho-ERK1/2 was determined with an ERK1/2 assay kit by immunopreciptation and Western blot analysis.

RESULTSThe expression levels of phospho-ERK1/2 kinase and the activity of phospho-ERK1/2 in the lung cancer cell line L9981-nm23-H1 were remarkably higher than those of the L9981 cell line and L9981-PLXSN cell line ( P < 0.01), but no significant difference in both the phospho-ERK1/2 expression and phospho-ERK1/2 activity was observed between the L9981 and L9981-PLXSN cell lines ( P > 0.05). There was no significant difference in the total ERK1/2 level among the three cell lines.

CONCLUSIONSnm23-H1 gene can obviously targetly suppress the activity of ERK1/2 in human high metastasis large cell lung cancer cell line L9981. This suggest that the mechanisms of nm23-H1 gene as a tumor metastasis suppressor gene may be related to its suppression to the MAPK/ERK signal transduction pathway.

BACKGROUNDIt has been known that oncogenesis and development of lung can- cer is a complex process regulated many genes and involved in multistages. Recent studies have demonstrated that signal transduction abnormality may play a very important role in these procedures. The aim of this study is to investigate the influence of exogenous MEK1/2 pathway inhibitor U0126 on the expression and activity of extracellular signal-regulated kinase (ERK1/2) of human high-metastatic large cell lung cancer cell line L9981 and its malignant biological behaviors.

METHODSThe expressive levels of total-ERK1/2, dually phosphorylated ERK1/2 and ERK1/2 relative activity of the human high-metastatic large cell lung cancer cell line L9981 (parent cell line with nm23-H1 gene hetero-deletion ) were detected by Western blot and immuno-precipitation technique after treating with different doses of U0126. The in vitro proliferative and invasive abilities of the lung cancer cell line were determined by MTT and modified Boyden chamber methods.

RESULTSThe ERK1/2 relative activity of L9981 gradually decreased accompanied with increase of U0126 doses, and a highly significant difference of phosphorylated ERK1/2 expression level existed among the different concentration groups of U0126 (P < 0.01), but no significant difference of total ERK1/2 expression was found among the different concentration groups of U0126 (P=0.387). After treatment with same concentration of U0126 for different time, the ERK1/2 relative activity of L9981 gradually decreased as the treatment time of U0126 prolonging, and a highly significant difference of phosphorylated ERK1/2 expression level was observed among different treatment time groups (P < 0.01). But no significant difference of total ERK1/2 expression level was found among different time groups (P=0.689). The inhibition of ERK1/2 pathway by MEK1/2 specific inhibitor U0126 targeting the ERK1/2 pathway of L9981 was dose- and time-dependent. After treatment with different concentration of U0126, the proliferation of L9981 gradually decreased accompanied with increase of U0126 concentration and a highly significant difference existed among different groups of U0126 concentration (P < 0.001). The in vitro invasion of L9981 gradually decreased accompanied with increase of U0126 concentration. No significant difference of in vitro invasion of L9981 was found among 0, 10 and 20μmol/l of U0126 (P > 0.05). A highly significant difference was observed when U0126 concentration increased to 40 and 60μmol/l compared with 0, 10 and 20μmol/l of U0126 (P < 0.001).

CONCLUSIONSThe inhibition of Ras-to-MAPK pathway by Ras-to-MAPK specific inhibitor U0126 targeting the Ras-to-MAPK pathway of the human high-metastatic large cell lung cancer cell line L9981 is dose-dependent and time-dependent. Suppressing or blocking of Ras-to-MAPK signal transduction pathway can reverse the invasive and metastatic phenotype of the human high-metastatic large cell lung cancer cell line L9981. These results suggest that the key kinase MEK1/2 of the ERK1/2 pathway may be a potent therapeutic target for human lung cancer.

BACKGROUNDnm23-H1 gene is a well-known tumor metastasis suppression gene. Our previous study has found that transfection of wild type nm23-H1 gene can significantly downregulate the ERK1/2 activity of human high-metastatic large cell lung cancer cell line L9981. The aim of this study is to investigate the influence of nm23-H1 and exogenous ERK1/2 pathway inhibitor U0126 on the extracellular signal-regulated kinase (ERK1/2) of human high-metastatic large cell lung cancer cell line L9981 and its malignant biological behaviors.

METHODSThe expressive levels of total-ERK1/2, dually phosphorylated ERK1/2 and ERK1/2 relative activity of the human high-metastatic large cell lung cancer cell lines, L9981 (parent cell line with nm23-H1 gene hetero-deletion), L9981-nm23-H1 (transfected with nm23-H1 gene ) and L9981-PLXSN (transfected with vector) were detected by Western blot and immunoprecipitation technique after treating with U0126 (40μmol/L for 20 minutes). The in vitro proliferative and invasive abilities among the above three lung cancer cell lines were determined by MTT and improved Boyden chamber methods.

RESULTSThe phosphorylated ERK1/2 expression level and relative activity in L9981-nm23-H1 lung cancer cell line were remarkably lower than those in L9981 and L9981-PLXSN lung cancer cell lines after being treated with U0126 (P < 0.01), but there was no significant difference between L9981 and L9981-PLXSN lung cancer cell lines. No significant difference of total ERK1/2 expression level was observed among the three lung cancer cell lines (P > 0.05) after being treated with U0126. The in vitro proliferation and invasion of L9981-nm-23H1 lung cancer cell line were remarkably lower than those of L9981 and L9981-PLXSN lung cancer cell lines (P < 0.01 ), but no significant difference was found between L9981 and L9981-PLXSN lung cancer cell lines (P > 0.05 ); U0126 could significantly down-regulate the in vitro proliferation and invasion of L9981 lung cancer cell line (P < 0.01).

CONCLUSIONSBlocking the activity of ERK1/2 in L9981 lung cancer cell line and transfecting the nm23-H1 gene into the L9981 lung cancer cell line may produce similar cell biological behavior changes, namely the significant reduction of in vitro proliferation and invasion of L9981 lung cancer cell line. These results indicate that the molecular mechanism which nm23-H1 gene reverses invasion and proliferation of the human high-metastatic large cell lung cancer cell line may be related to its effects of down-regulating the activity of the key kinase ERK1/2 of Ras-to-MAPK signal transduction pathway.