50 000 bp)was extracted from frozen gastric cancer tissue and their corresponding normal samples and used for RLGS assay.The genome DNA was digested by methylation-sensitive restriction enzyme Not Ⅰ, and labeled by radioisotope ~(32)p,then separated by two-dimensional electrophoresis and autoradiography. Experimental conditions for each step were optimized step by step.DNA fragment sequences for the dots on scanning profile were identified according to the human RLGS database.Results RLGS assay was set up and used to get the RLGS profiles of the representative tested samples successfully.These profiles can display more than 1 200 available dots averagely,the profiles of high quality DNA sample can display more than 1 800 dots which is the average level at an excellent RLGS lab,discrepant dots which had weaken or enhanced signals and their sequence information were obtained.The result can be reproduced.Conclusion The RLGS assay is established,stabilized for detection of DNA methylation of tissue samples.

Objective To investigate the distribution of acquired carbapenemases in carbapenemresistant strains of Klebsiella pneumoniae, and explore its role in epidemiology of nosocomial infection. Methods From November 2008 to March 2009, twenty clinical isolates of carbapenem-resistant Klebsiella pneumoniae were collected from children hospitalized in Wuhan children's hospital. MICs of antibiotics were tested by DNA of Klebsiella pneumoniae. Modified Hodge test was used to screen strains producing carbapenemases,combined imipenem(IPM)-EDTA , meropenem(MEM)- EDTA and ceftazidime(CAZ) - EDTA double-disk synergy test (DDST) were used to detect metallo-β-lactamase-producing. PCR amplification of the carbapenemase and integrase genes, and sequencing were performed. Plasmid conjugation transfer experiments and Southern hybridization were applied to study the mode of drug resistance transmission. Results Four types of Klebsiella pneumoniae were detected by PFGE, type A consisted of 5 strains, including 3 strains of type Al and 2 strains of type A2), type B (2 strains), type C (12strains) and type D (1 strain). Type A and C were the main drug resistant clones. Eight strains of Klebsiella pneumoniae carried both KPC-2 and IMP-4 genes, 10 strains carried IMP-4 gene, 2 strains carried KPC-2 gene. None of NDM-1 ,GIM, SPM, SIM, OXA-23, and VIM carbapenemase genes was detected in 20 isolates. All of 20 isolates carried lntl which were found to be located on bacterial chromosome by Southern blot. Conclusions KPC-2and IMP-4 genes are the major carbapenemase genes in Klebsiella pneumoniae isolated in Wuhan.Transmission of drug resistance is mainly through vertical transmission of type C resistant clone and horizontal transmission of Intl on bacteria chromosome.

Objective To investigate the molecular epidemiology of CRAB isolated from children in wuhan. Methods Forty non-repetitive strains of CRAB were collected from hospitalized children of emergency department, neonatal medicine, cardiothoracic surgery, bone surgery, respiratory medicine and renal medicine in Wuhan children's hospital during December 2008 and May 2009. MIC values were PFGE; KPC, IMP, GIM, SPM, SIM, OXA-23, VIM genes and integrase gene were amplified by PCR and then sequenced to confirm the genotypes.; Plasmid conjugation experiment was used to study the transfer method of bacterial resistance and southern blot hybridization was used to target the resistance genes. Results Susceptible rates of 40 strains to gentamicin, tobramycin, amikacin, ciprofloxacin, levofloxacin, trimoxazole were 20%, 5%, 93%, 93%, 95%, and 23% respectively. Eleven types of clone were detected by PFGE,including 29 strains of type A clone, 2 strains of type B clone, and 1 strain for each type of C to K clone. Eleven isolates produced both IMP-4 and OXA-23 carbapemase. Twenty-six isolates only possessed OXA-23 carbapemase. Thirty-six strains carried class Ⅰ integron. The results of southern blot hybridization showed that Intl, IMP-4 and OXA-23 type were located on chromosome. Conclusions Type A clone of CRAB is the most common. OXA-23 and IMP-4 type are the major acquired carbapemases, especially the OXA-23 is the most common type. The horizontal transmission of OXA-23 and IMP-4 gene mediated by Int1 and the spread of type A resistant clone is the major way of the spread of carbapenem-resistant Acinetobacter baumannii in the region.

Data quality management system is essential to ensure accurate, complete, consistent, and reliable data collection in clinical research. This paper is devoted to various choices of data quality metrics. They are categorized by study status, e.g. study start up, conduct, and close-out. In each category, metrics for different purposes are listed according to ALCOA+ principles such us completeness, accuracy, timeliness, traceability, etc. Some general quality metrics frequently used are also introduced. This paper contains detail information as much as possible to each metric by providing definition, purpose, evaluation, referenced benchmark, and recommended targets in favor of real practice. It is important that sponsors and data management service providers establish a robust integrated clinical trial data quality management system to ensure sustainable high quality of clinical trial deliverables. It will also support enterprise level of data evaluation and bench marking the quality of data across projects, sponsors, data management service providers by using objective metrics from the real clinical trials. We hope this will be a significant input to accelerate the improvement of clinical trial data quality in the industry.

OBJECTIVETo observe the clinical effect of Huikangling Tablet (HT, extracted from Scabrous Patrinia root) on peripheral blood micrometastasis of differentiated thyroid carcinoma (DTC) patients.

METHODSTotally 87 DTC patients with positive micrometastasis were randomly assigned to the treatment group (45 cases) and the control group (42 cases). DTC endocrine inhibition treatment standards were executed in all patients. They all took levothyroxine sodium (50 microg/tablet, from low dose, 25 microg each time, once per day, 0.5 h before breakfast), and its dosage was gradually added one week later. The dosage was adjusted according to tested results of TSH combined recurrence risk stratification and endocrine suppression induced adverse reactions risk stratification. Patients in the treatment group took HT (0.4 g per tablet, 3 tablets each time, three times per day for a total of 12 weeks) combined TSH suppression therapy, while those in the control group only received TSH suppression therapy. Peripheral micrometastatic cytokeratin 19 (CK19) and polymorphic epithelial mucin1 (MUC1) were detected by FCM at week 4 and 12. Meanwhile, distant metastasis and adverse reactions were observed.

RESULTSAfter 4-week treatment positive micrometastasis was shown in 18 cases (40%) of the treatment group and 29 cases (69%) in the control group with statistical difference (chi2 = 5.68, P < 0.05). After 12-week treatment positive micrometastasis was shown in 7 cases (15.6%) of the treatment group and 17 cases (44.7%) in the control group with statistical difference (chi2 = 8.49, P < 0.01). Pulmonary metastasis occurred in 2 cases and bone metastasis in 1 case of the control group at follow-ups. Cervical lymph node metastasis without accompanied recurrence of thyroid cancer occurred in one case of the treatment group. No obvious liver or renal abnormalities occurred.

CONCLUSIONHT inhibited peripheral blood micrometastasis of DTC patients and its mechanism needed to be further studied.

Antineoplastic Agents ; pharmacology ; therapeutic use ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Neoplasm Micrometastasis ; drug therapy ; Neoplasm Recurrence, Local ; Tablets ; Thyroid Neoplasms ; drug therapy

Purpose To investigate the expression of polo-like kinase 1 (Plk1) and Cyclin B1, p21WAF1in cervical carcinoma, and to determine the relationship between the expression of the three proteins and tumor clinicopathological features. Methods The expres-sion of Plk1, Cyclin B1 and p21WAF1 was detected in 102 cases of cervical carcinoma, 20 cases of (cervical intraepithelial neoplasia, CIN) , and 20 cases of nomal cervical tissues by the technique of tissue chip and immunohistochemical staining of EliVision. Statistical analyses of the data were performed with SPSS 19. 0 software. Results The positive rates of Plk1 in cervical carcinoma and CIN were 70. 5%, 55. 0%, respectively, which were significantly higher than normal cervical tissues (10%) (P<0. 01);The positive rates of Cyclin B1 in cervical carcinoma and CIN were 52. 9% and 30. 0%, respectively, which were significantly higher than normal cervical tissues (10%)(P <0.01); The positive rates of p21WAF1 in cervical carcinoma and CIN were 23.5% and 10.0%, respectively, which were significantly higher than normal cervical tissues ( 0 ) ( P<0. 01 ) . There were no significant differences between cervical carcinoma and CIN in the positive rates of Plk1, Cyclin B1 and p21WAF1. The expression of Plk1 was associated with the depth of carci-noma invasion (P<0. 05), that of Cyclin B1 was associated with lymph node metastases and the depth of carcinoma invasion (P<0. 05)and that of p21WAF1 in cervical carcinoma was associated with histological grade (P<0. 05). In cervical carcinoma, the expres-sion of Plk1 was positively correlated with Cyclin B1 (rs =0. 297, P=0. 002) and negatively correlated with p21WAF1(rs = -0. 403, P<0. 001). Conclusion The expression of Plk1, Cyclin B1 and p21WAF1 is involved in the occurrence and development of cervical carcinoma, and the former two are also related with prognosis of cervical carcinoma. The combination of the three would provide a new target for clinical treatment.

Objective To investigate the effects of intestinal ischemia reperfusion (IIR) on the progression of inflammatory reaction in hemorrhagic necrosis pancreatitis (HNP).Methods Eighty rats were randomly divided into sham operation (SO) group,acute edematous pancreatitis (AEP) group,AEP + IIR group and HNP group according to the random number table.Erythrocyte velocity (EV),functional capillary density (FCD) and leukocyte adherence (LA) were observed at 0,1,2,3 and 6 hours after the models were completed.The serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected.All data were analyzed by using the analysis of variance or the t test.Results The level of EV in the AEP group significantly decreased at 1 hour,and got increased at 3 hours,while the level of EV in the AEP group was still significantly lower than that in the SO group ( t =9.60,P ＜ 0.05 ).The levels of EV in the AEP + IIR group and HNP group constantly decreased,and increased at 6 hours,but were continually lower than that in the AEP group ( t =6.03,6.12,P ＜0.05 ).The level of FCD in the AEP group was significantly lower than that in the SO group at 3 hours ( t =8.20,P＜0.05).The levels of FCD in the AEP + IIR group and HNP group were significantly lower than that in the AEP group at 3 hours (t =35.60,23.80,P ＜ 0.05 ).Compared with AEP group,the level of LA in the AEP group was significantly increased at 1 hour ( t =75.00,P ＜ 0.05 ) and reached peak at 3 hours.The levels of LA in the AEP + IIR group and HNP group were significantly higher than that in the AEP group at 1,2,3,6 hours (t =23.00,29.50,53.00,38.70,23.10,48.20,39.20,47.50,P＜0.05).Compared with SO group,the level of TNF-α in the AEP group significantly increased since l hour (t =77.00,P ＜ 0.05),and began to decrease at 3 hours; the levels of TNF-α in the AEP +IIR group and HNP group at 2 hours were significantly higher than that in the AEP group (t =23.50,18.10,P＜0.05).The levels of IL-6 in the AEP group at 1,2,3,6 hours were significantly higher than those in the SO group ( t =93.50,146.00,243.60,209.20,P ＜ 0.05 ).The levels of IL-6 in the AEP + IIR group and HNP group at 1 hour were not significantly different from that in the AEP group ( t =2.30,2.03,P ＞ 0.05),while the levels of IL-6 in the AEP + IIR group and HNP group at 2 hours were significantly higher than that in the AEP group (t =35.63,29.80,P ＜ 0.05 ).Conclusion IIR may enhance the inflammatory reaction of AEP and IIR might be one of the factors to exaggerate the inflammatory reaction of HNP.

Objective To explore the prognostic impact of miR137 target gene Kruppel-like transcription factor 12 (KLF12) in multiple myeloma (MM). Methods The target genes of miR137 were predicted by software. The GFP analysis of KLF12 and the prognosis of MM were constructed. Overexpressing miR137 in MM NCI-H929 cell line was also constructed. Real-time qPCR and Western blot were used to detect the expression of KLF12 in this cell line. Results The target genes of miR137 were MITF, BUE2H, SH3BP5 and KLF12. High expression of KLF12 in 455 patients included 75 patients (16.5 %) died, 104 patients with low expression of KLF12, and 25 patients (24.0 %) died, but no significance was detected in the different subgroups. KLF12 expression was higher in MM NCI-H929 cell line with miR137 over expression. The expression of miR137 was positively correlated with the expression of KLF12. Conclusion miR137-KLF12 is an important index to judge the prognosis of MM.

Objective To investigate the clinicopathological features,diagnosis,treatment,prognosis and causative agent of a case of eumycetoma on the submaxilla.Methods A case of eumycetoma diagnosed in our department was assessed for its clinical and pathological features as well as mycologic and molecular identification.Related literature was reviewed.Results The patient was primarily characterized by swelling of the submaxilla,with multiple sinuses draining many black granules.Pathologic examination revealed a pyogenic granulomatous inflammation,and a number of lotus rhizome node-like hypha were observed in tissue samples through PAS staining.Sequence analysis of multiple loci of the isolates,including ITS 1,ITS2 and D1/D2,showed that it was mostly similar to Madurella mycetomatis with a homology of 97%.Conclusion This is a case ofeumycetoma on the submaxilla induced by a novel species of Madurella.

AIM:To study the effect of posterior sub - Tenon's capsule injection of triamcinolone acetonide ( TA ) in treatment of patients with diffuse diabetic macular edema ( DME) before panretinal photocoagulation ( PRP) .
METHODS:Retrospective analysis of the clinical data of 96 cases (96 eyes) with DME treated in our hospital from October 2008 to May 2012, and the patients were divided into the study group and control group, each group with 48 cases ( 48 eyes ) , the control group were only treated with PRP, and for the study group, TA was injected one week before PRP. At 6mo after treatment, best-corrected visual acuity ( BCVA) and retinal thickness changes of two groups were compared, the changes of intraocular pressure in two groups was analyzed.
RESULTS:After treatment, two groups were followed up for 6mo, compared with before treatment, the expression of BCVA in control group was reduced, and rise in the study group, with significant difference between the two groups (P<0. 05), and during the follow-up period, IOP change was in the normal range for the two groups, with no the difference (P>0. 05), the study group had foveal thickness reduction of 9. 6μm, the control group was increased by 31. 9μm, with significant difference (P<0. 05), parafoveal thickness in the study group decreased 5μm, significantly increased 22. 1μm in the study group, centre concave surrounding thickness increased 0. 4μm in study group and 19. 4μm for the control group, with no significant difference (P>0. 05).CONCLUSION:TA injection in patients with diffuse DME before PRP is safe and effective, and it is superior to simple PRP therapy, and it can be applied in primary hospital.