In this study,the loop-mediated isothermal amplification(LAMP)method was used to develop a rapid and simple detection system for porcine circovirus type 2(PCV2).According to the PCV2 sequences published in GenBank,multiple LAMP primers were designed targeting conserved sequences of PCV2.Using the DNA extracted from PCV2 isolates HUN-09 and SD-09 as the template,LAMP reactions in a PCV2 LAMP system was performed,the amplification products were detected by adding SYBR Green I and could be observed directly by the naked eye.The results showed highly-efficient and specific amplification in 30 min at 63℃ with a LAMP real-time turbidimeter.Furthermore,PCV2 DNA templates,with a detection limit of 5.5×10-5ng of nucleic acid,indicated that this assay was highly sensitive.The results obtained with the naked eye after SYBR Green I staining were consistent with those detected by the real-time turbidimeter,showing the potential simplicity of interpretation of the assay results.The LAMP assay appeared to have greater accuracy than PCR and virus isolation for the analysis of 18 clinical samples.In addition it offers higher specificity and sensitivity,shorter reaction times and simpler procedures than the currently available methods of PCV2 detection.It is therefore a promising tool for the effective and efficient detection of PCV2.

Objective: To introduce electroacupuncture (EA) treatment on cervical spondylosis radiculopathy (CSR) at home and abroad in the prior ten years. Methods: EA, combined EA and other treatments for NCS are classified and analyzed. Results: The most commonly used acupoints are JiaJi (Ex-B2), Fengchi (GB 20), Waiguan (TE 5), Quchi (LI 11). EA is widely applied to treat CSR because of its definite therapeutic effects and safety. Conclusion: The validity of other evaluation indexes needs to be proved by more rigorously designed randomized controlled trials (RCT) with scientific methods.

Objective To study a series of tests to the IMRT planning system RayStation,and verify the feasibility of this Standard.Referring to the standard Performance and test methods for intensity modulated radiation therapy (IMRT) treatment planning system (Standard).Methods Referring to the Standard,the tests include:dose goals used for planning,point dose calculation accuracy and dose distribution calculation accuracy.And test phantoms include:AAPM TG119 report's test cases and solid water phantom for verification,and IMRT test cases include:multitarget,mock prostate,head/neck and CTshape (easier version and harder version) and so on.Referring to the Standard,we optimize and calculate doses of interest in RayStation,and use the common measurement equipments and tools,such as an ionization chamber,films,a detector array,and etc,to measure the doses on the accelerator,then calculate and analyze the errors between them.Results The results of dose goals for five different mock plans meet the requirements of the Standard.And the point dose ionization chamber measurements are in line with the requirements of the Standard,and the total error of the mock plans was (0.83 ± 1.65) ％.The γpass rates of per-field measurements using detector array are all greater than 99.0％,and the γpass rates of composite film measurements are all greater than 92.5％.Conclusions The results of three trials to IMRT performance of RayStation are in full compliance with the requirements of the Standard.And also it is an evidence for the feasibility of this Standard.

Objective To understand laboratory diagnosis and clinical characteristics of human immunodeficiency virus (HIV ) infection and AIDS patients in three comprehensive hospitals .Methods Laboratory diagnosis and clinical characteristics dates of people living with HIV/AIDS patients consulted in Chongqing Emergency Medical Center between 2007 to 2013 were retrospective‐ly analyzed .Results Totally 47 355 cases were carried HIV antibody screening during 7 years ,179 cases of HIV antibody were positive in preliminary screening ,171 cases were confirmed as positive .Among 5 cases of HIV antibodies result unconfirmed ,2 ca‐ses were followed up ,1 case was ruled out HIV infection ,1 case was converted to HIV antibody positive .People living with HIV al‐ways merging double or multiple infection with hepatitis b virus (HBV) ,hepatitis c virus (HCV) and treponema pallidum (TP) and so on .People living with HIV aged from 18 to 86 years old ,9 .36% was over 60 years old .Most patient has two or more clinical manifestations when consulted a doctor .Conclusion There were false‐positive of HIV antibody preliminary screening ,HIV anti‐body positive results must be confirmed by Western blot confirmatory test .Uncertainty of HIV antibody results should be judged by regularly follow‐up or combining with other detection methods ,epidemiological data .Routine HIV antibody screening should be adopt for HBV ,HCV and TP infection .Elder patients should not be ignored .Clinical specificity of HIV/AIDS is not strong ,it is need to be valued and identified from other cause similar symptoms of diseases caused by phase identification ,in order to reduce missed diagnosis and misdiagnosis .

Rhamnolipid biosurfactant is mainly produced by Pseudomonas aeruginosa that is the opportunistic pathogenic strain and not suitable for future industrial development. In order to develop a relatively safe microbial strain for the production of rhamnolipid biosurfactant, we constructed engineered Escherichia coli strains for rhamnolipid production by expressing different copy numbers of rhamnosyltransferase (rhlAB) gene with the constitutive synthetic promoters of different strengths in E. coli ATCC 8739. We further studied the combinatorial regulation of rhlAB gene and rhaBDAC gene cluster for dTDP-1-rhamnose biosynthesis with different synthetic promoters, and obtained the best engineered strain-E. coli TIB-RAB226. Through the optimization of culture temperature, the titer of rhamnolipd reached 124.3 mg/L, 1.17 fold higher than that under the original condition. Fed-batch fermentation further improved the production of rhamnolipid and the titer reached the highest 209.2 mg/L within 12 h. High performance liquid chromatography-mass spectrometry (LC-MS) analysis showed that there are total 5 mono-rhamnolipid congeners with different nuclear mass ratio and relative abundance. This study laid foundation for heterologous biosynthesis of rhanomilipd.
Bacterial Proteins ; genetics ; Batch Cell Culture Techniques ; Decanoates ; Escherichia coli ; metabolism ; Fermentation ; Glycolipids ; biosynthesis ; Hexosyltransferases ; genetics ; Industrial Microbiology ; methods ; Multigene Family ; Promoter Regions, Genetic ; Pseudomonas aeruginosa ; Rhamnose ; analogs & derivatives ; biosynthesis ; Surface-Active Agents ; metabolism

Objective To compare the dose to the contralateral breast, ipsilateral lung, and the whole lung in the tangential field radiotherapy for primary breast cancer using the dynamic wedge or physical wedge. Methods With thirteen breast cancer patients chosen, the dose distribution was computed for the plan used in practical treatment with the dynamic wedge and physical wedge. Plans were compared using dose volume histograms for the contralateral breast, ipsilateral lung and the whole lung. As for the contralateral breast, the dose distributions were not computed for the whole breast but computed for the two regions similar to a rectangular area in the axial slice and parts of the whole breast. The mean dose was used to evaluate CB1, CB2 and ipsilateral lung, and V_ 20 was used to evaluate the whole lung. The treatment planning system used was Varian CadPlan. An ionization chamber in a water phantom was used to measure some point doses to simulate the dose to the contralateral breast. Results When using the 30? dynamic wedge, the mean dose to CB1 and CB2 was 1.5%-3.9% and 1.1%- 2.6% , and the mean dose to the ipsilateral lung was 4.1%-14.7%. When using the 30? physical wedge, the mean dose to CB1 and CB2 were 1.5%-4.4% and 1.2%-3.0%, respectively, and the mean dose to the ipsilateral lung was 4.4%-15.2%. The values of V_ 20 were equal. When using the 15? dynamic wedge, the mean dose to CB1 and CB2 decreased compared to 15? physical wedge, but the value reduced was smaller than when using 30? wedge. Also, the measured results verified that the dose to the normal tissue is reduced using the dynamic wedge. Conclusions The mean dose to the contralateral breast was reduced by using the dynamic wedge instead of the physical wedge, and the mean dose to the ipsilateral lung or V_ 20 is reduced or equal to each other. So the probability of normal tissue complication such as a second breast malignancy or pneumonitis associated with radiotherapy are likely to be reduced.

Objective To construct DNA damage and repair gene apurinic/apyrimidinic endonuclease (APE1) siRNA expression vector pSilence APE1 and investigate its inhibitory effect on the expression of APE1 in osteosarcoma cell HOS and 9901 in vitro. Methods An expression plasmid of a short hairpin RNA target APE1, pSilence APE1 was constructed and transfected to 9901 and HOS cell by lipofectamine. The expression of APE1 protein in HOS and 9901 osteosarcoma cells posttransfected with pSilence APE1 was detected by immunohistochemistry. Meanwhile, the dose-effect and time-effect relationship of APE1 gene silence induced by pSilence APE1 were measured using Western blot analysis. Results After evaluation and sequencing, the APE1 siRNA expression vector pSilence APE1 was constructed successfully. The result of Western blotting and immunohistochemistry showed that APE1 protein in osteosarcoma cells could be knocked down specifically by pSilence APE1, and the inhibition rate of APE1 expression was 72%-95%. The best inhibition of expression of APE1 gene was 3.0 ?g and at the 72 h using pSilence APE1. Conclusion APE1 siRNA expression vector pSilence APE1 is successfully constructed that can significantly knock down APE1 gene expression in osteosarcoma cells.

Objective To study the expressions and correlation of cyclooxygenase-2 (COX-2) and epidermal growth factor receptor (EGFR) in human transitional cell carcinoma of bladder (TCC) tissues and explore the correlation of their expressions in the incidence and development of TCC. MethodsThe expressions of COX-2 and EGFR were detected by immunohistochemical technique in 48 cases of TCC and in 10 cases of normal bladder mucosa tissues. ResultsThe expressions of COX-2 and EGFR in TCC was significantly higher than those in normal bladder mucosa tissues (P

Objective To detect the expression of apurinic/apyrimidinic endonuclease 1 (APEI) and explore its correlation with the expression of mutant p53 in hepatocellular carcinoma (HCC). Methods The expression of APE1 and mutant p53 was detected by SP immunohistochemical method in 10 specimens of normal liver tissue, 40 specimens of liver cirrhosis tissue and 103 specimens of HCC tissue which were collected at the Department of Pathology of Daping Hospital from 1991 to 2004. All data were analyzed by chi-square test, correla-tion analysis and K Independent-Samples Tests. Results The expression rate of APE1 in HCC was 100.0%, which was significantly higher than that in normal liver tissue (40.0%) and liver cirrhosis tissue (82.5%) (χ~2= 47.852, P < 0.01). The expression of APE1 was only detected in the nucleus in normal liver tissue. Ectopic expression of APE1 in cytoplasm was detected in liver cirrhosis tissue and HCC tissue, with the rate of 20.0% and 53.4%, respectively (χ~2=20.757, P <0.01). There was statistical difference in clinical staging and pathological grading of HCC with different combinations of APE1 expression (intranuclear or ectopic expression) and mutant p53 expression (positive or negative expression) (χ~2=12.910, 14.481, P < 0.01), and HCC with ectopic expression of APE1 and positive expression of p53 had high malignant degree. Conclusion Overexpression and ectopic expression of APE1 in cytoplasm may play important roles in the genesis and progression of HCC, and the ectopic expression of APE1 and p53 mutation may have synergistic effect.

Objective To investigate the value of combined detection of multi-tumor markers in the diagnosis of primary hepatocellular carcinoma (HCC) and to establish the discriminant equation. Methods Using a protein chip, 12 tumor markers in the serum from 98 patients with HCC and 67 patients with benign liver diseasewho had been admitted to Daping Hospital from November 2003 to April 2006, and 46 healthy individuals during he same period were analyzed. A discriminant equation was established to discriminate primary HCC from benign liver diseases. All the data were processed by variance analysis and chi-square test. Results The positive rates of the tumor markers were 89% (87/98) in patients with primary HCC, 19% (13/67) in patients with benign liver disease and 4% (2/46) in healthy individuals. There was statistical difference in the serum level of alpha-fetoprotein (AFP), eareinoembryonic antigen (CEA), ferritin (FER), CA19-9 and CA125 among the 3 groups (F =59.530, 40.472, 31.708, 75. 897, 153.066, P <0.05). Combined detection of AFP, CEA, FER, CA19-9 and CA125 improved the diagnostic accordance rate to 89%, which was significandy higher than the diagnostic accordance rate (64%) when only AFP was detected (X2 = 16.362, P <0.05). The accuracy of the discriminant equation was 90%. Conclusions Combined detection of multi-tumor markers is superior to AFP detection. Combined detection of multi-tumor markers can be used in screening of the HCC patients in HCC high risk population and in the early diagnosis of primary HCC.