Objective In this research with the method of Alu -PCR we investigate the integration of hepatitis B virus ( HBV) DNA in human hepatocarcinoma cell line (PLC/PRF/5) genome DNA.Methods We at first extracted the genome DNA from PLC/PRF/5 cells, and then the potential integration fragments of HBV DNA and human genome DNA were amplified with according to the Alu -PCR after three rounds PCR .The Alu-PCR amplification products were observated with agarose gel electrophoresis , then integration positive electrophoresis bandings were chipped and purified for nucleic acid sequencing .At last he bioinformatics information was acquired by blast online.Results Through agarose gel electrophoresis after Alu -PCR amplification, we got four potential integration bindings , among which we got three integration sequences of HBV DNA in human genome DNA .These integration sequences could be individually located in the human chromosome of 03p21.31, 05p15.33, 12q13 and 12-q14.1.Conclusion With Alu-PCR we can accurately measure the integration of HBV DNA in human genome DNA , and Alu-PCR can be a a convenience and economic method in the study of HBV DNA ′s integration in human genome DNA .

OBJECTIVE To establish a rapid detecting method for Clostridium perfringens on traumatic tissues by type-specific multiplex PCR.METHODS Established a simple and rapid method(TLS method) for purifying the DNA of genomes and plasmids in standard strains of C.perfringens wild strains on surface of open traumatic tissues and detected DNAs by type-specific multiplex PCR.RESULTS All types of C.perfringens could be detected by type-specific multiplex PCR.The sensitivity by PCR for type A of C.perfringens arrived at 7.5?103/ml and a whole run could finish within 5 h;the results by PCR entirely corresponded with those by cultureing.CONCLUSIONS New methods for purifying DNA of genomes and plasmids of C.perfringens are simple and rapid;there are high specificity and sensitivity for detecting DNA by multiplex PCR within short time,which can be practiced in clinical laboratory.

Objective To study the effectiveness of the laboratory diagnosis of multiple myeloma(MM)patients with immun-ofixtion electrophoresis (IFE),protein electrophoresis (SPE)and immunoglobulins and light chain quantitative analysis. Methods Selected 192 MM patients and 30 healthy controls during June 2012 to December 2013 and analyzed the results of IFE,SPE and immunoglobulins,and light chain quantitative analysis in MM patients.Results M protein bands were seen in 120 cases (62.5%)by using SPE and M protein were positive in 174 cases (90.6%)among the 192 MM patients by using IFE.IFE showed that IgG were maximum type of the M protein (106 of 192,55.2%).There were IgG-λ type 66 cases (34.4%),IgA type 36 cases (18.8%)and free light chain type 24 cases (12.5%).Immunoglobulins of different immuno-phenotypes had higher than the nomal group with serum immunoglobulin and light chain quantitative analysis (P <0.05). The detection rate was 67.7% (130/192).Whateverκ-type M protein orλ-type M protein,the ratio ofκ/λwas significantly abnormal (P <0.05).The detection rate was 85.4% (164/192).Conclusion The better detection rate of immunological techniques such as immunofixtion electrophoresis and immunoglobulins quantitative analysis might provide valuable basis for the diagnosis and treatment of MM clinically and prevent misdiagnosis.

Objective To investigate the clinical significance of homocysteine(Hcy) and glycated hemoglobin (HbA1c) in patients with diabetic retinopathy.Methods From March 2013 to February 2015,a total of 168 patients with diabetes from Beijing Jishuitern Hospital were divided into diabetic retinopathy group (76 cases,32 male,44 female,average age 62.97 ± 12.46) and non-diabetic retinopathy group (92 cases,46male,46 female,average age 58.14 ± 13.71),and 65 cases of healthy subjects (46male,46 female,average age 60.24 ± 10.85) were enrolled as normal control group as well.Hcy and HbA1c were detected in all groups.Multiple independent samples nonparametric tests were used to compare the difference between groups.The concentrations of Hcy and HbA1c between DR group and NDR group were analysised by ROC curve analysis.Result The Hcy levels in normal control group,NDR group and DR group were 7.00 (4.55-10.50) μmol/L,12.45 (10.30-15.10) μmol/L,20.10 (16.63-25.68) μmol/L respectively.The HbA1c levels in normal control group,NDR group and DR group were 5.30 (5.00-5.70) ％,6.80 (5.83-8.03) ％,7.25 (6.50-8.60) ％ respectively.There was no significant difference in Hay and HbA1c levels between groups (P ＜ 0.01);Compared with non-diabetic retinopathy group,levels of Hcy,HbA1c in diabetic retinopathy group were obviously higher;The optimal diagnostic cut-off values of Hcy was 14.35 μmol/L,at this cut-off value,the sensitivity was 96.05％,specificity was 68.48％;The optimal diagnostic cut-off values of HbA1c was 6.05％,at this cut-off value,the sensitivity was 98.68％,specificity was31.25％.Conclusion Patients with Diabetic retinopathy might have hyperhomocystinemia,and detection of Hcy may contribute to the early diagnosis of diabetic retinopathy.

Treatment based on syndrome differentiation is an essential feature of traditional Chinese medical diagnosis. The interventions based on changes of syndrome types in randomized controlled trials are complicated, leading to the difficulty of blind method enforcement. This article described a double-blind method. It could be used in randomized controlled trials under the condition of different syndrome types and different medications. It numbered drugs in two stages, and in two phases to achieve double-blind. This method not only guaranteed investigators and subjects to be in blinded conditions, but also achieved using different medications for patients of different syndromes. It also caused no drug waste. It was scientific and feasible.
Double-Blind Method ; Humans ; Medicine, Chinese Traditional ; Randomized Controlled Trials as Topic ; Single-Blind Method

OBJECTIVE To explore the risk factors of gas gangrene after open trauma for its early diagnosis and precaution.METHODS The data of 42 patients with gas gangrene after open trauma from Aug 2003 to Aug 2008,were collected.The risk factors of gas gangrene after open trauma had been retrospectively analyzed from the cause of the injuries,the distribution of latent period,wound,systemic symptoms,laboratory examination and treatment.RESULTS Gas gangrene after injury was related with open wounds for various reasons.Latent period of gas gangrene was 24-72 h.Early risk factors of gas gangrene among them mainly included degloved or avulsed skin-injury(100%),musce trauma(100.00%),contaminated wound and retained foreign bodies(95.24%),weak pulse of peripheral artery(95.24%),nervous injury(90.48%),and lower temperature of skin aound the wound(83.33%).There were obviously systemic symptoms in infective stage but in early stage were not.CONCLUSIONS Gas gangrene is extremely rare in injury patients but it is life-threatening.So more attention should be paid to the risk factors in order to decrease the incidence of gas gangrene after trauma.Improving the prognosis of patients;the wounded locations should be effectively treated early so as to improve the prognosis.

Objective To evaluate the diagnostic value of the metabolic parameters for differentiating focal autoimmune pancreatitis (F-AIP) and pancreatic cancer (PC) by dual time 18F-FDG PET/CT scan.Methods Ten F-AIP patients and 20 PC patients in Changhai Hospital from May 2011 to November 2014 were enrolled in this study.All the AIP patients were histological confirmed or diagnosed by clinical follow up.The PC patients were histological confirmed and gender-and age-matched with F-AIP patients.50％ SUVmax was set as the threshold to fine-tune the boundary of interest.The extracted parameters included SUV SUV metabolic tumor volume (MTV),total lesion glycolysis (TLG),target-to-background ratio (TBR) and the retention indexes(RI) of all the parameters above.The PET/CT imaging features were also observed.Results The high metabolic lesions were observed in both F-AIP patients and PC patients.There were 6 F-AIP patients whose lesion was located in pancreas head,4 F-AIP patients whose lesion was located in pancreas body and tail.There were 12 PC patients whose lesion was located in pancreas head,8 PC patients whose lesion was located in pancreas body and tail.In F-AIP patients,2 cases had dilated pancreatic duct,6 had dilated biliary duct,8 had increased metabolism in mediastinal lymph node and 2 had abdominal lymphadenopathy,which were 8,5,5 and 14 cases in PC patients.The positive rate of mdeiastinal lymphadenopathy in F-AIP patients was statistically higher than that in PC patients,while the positivity rate of abdominal lymphadenopathy in AIP patients was lower than that in PC patients.The difference was statistically significant (both P ＜ 0.05).There were no statistical differences on the positivity rate of the dilated pancreatic duct,intra-and extra-hepatic bile duct between two groups.SUVmax,SUVmean and MTV in F-AIP were 5.37 ± 0.88,3.48 ± 0.66,21.79 ±15.60 in early stage and 6.45 ±1.51,4.23 ± 1.10,19.36 ± 14.63 in delayed stage,and those in PC were 8.31 ±3.08,5.41±1.95,9.26±8.35 in early stage,and 9.75±3.86,6.36±2.56,9.09±10.71 in delayed stage.SUVmax and SUVmean in F-AIP were lower than those in PC,whereas MTV were larger in F-AIP than that in PC.ROC curves for SUVmax,SUVmean and MTV were made.The AUC of SUV was the highest at 0.85,the cut-off value was 4.45,the corresponding sensitivity was 65％ and the specificity was 90％.TLG,TBR and RI of all the parameters were not statistically different in F-AIP and PC.Conclusions The 18F-FDG PET/CT metabolic parameters,such as SUVmax,SUVmean,MTV,could be of special diagnostic significance in discriminating F-AIP from PC.

Objective To investigate the dynamic changes of dengue viral loads and IgM antibody in patients with dengue fever ( DF) and to analyze their relationships with disease severity. Methods A to-tal of 1 508 serum samples were collected from 1 140 hospitalized patients including 1 050 mild cases and 90 severe cases within 10 days after the onset of DF in Guangzhou in 2014. The viral loads were determined by using fluorescence quantitative RT-PCR. ELISA was performed to measure the dengue virus ( DENV)-spe-cific IgM antibody. Results In general, the DENV viral loads in patients declined gradually from 108 copies/ml on day 1 to 103 copies/ml on day 10 after the onset of DF. The viral loads in severe cases were significantly higher than those in mild cases on days 5 to 7 (P<0. 05). The positive rates of DENV RNA in serum samples also decreased with the disease progression from 100% on day 1 to 40% on day 10. Com-pared with the mild cases, the patients with severe DF showed higher positive rates of DENV RNA on day 6 and day 8 (P<0. 05). The DENV-specific IgM antibody could be detected on day 2 and the secretion of IgM antibody increased gradually with the disease progression. The levels of IgM antibody in mild cases were sig-nificantly higher than those in severe cases (P<0. 001). The positive rates of IgM antibody in patients in-creased form 8% on day 2 to 95% on day 6. Higher positive rates of IgM antibody were detected in mild ca-ses on days 5 and 6 as compared with those in patients with severe DF (P<0. 05). Conclusion High viral load and low level of IgM antibody during the fastigium of DF were closely associated with the disease severity.

Objective To explore the status and related influencing factors of disaster preparedness for ICU nurses and provide reference for carrying out learning and training of the knowledge of specialized disaster rescue. Methods With convenience sampling of network questionnaire, ICU nurses in two Fujian province hospitals attended the investigation. Results A total of 347 valid questionnaires were recovered. The score of ICU nurses disaster preparedness score was 146.32 ± 38.71. located in the middle level. Sex, working years, educational background, disaster relief experience, disaster knowledge and skill training were the main factors influencing the disaster preparedness of ICU nurses (t=-4.190-2.682, P＜0.01). Conclusions The level of disaster preparedness degree for ICU nurses was not optimistic. It was necessary to actively formulate and improve targeted training of disaster nursing theory knowledge and disaster nursing skills drills, so as to effectively promote the disaster preparedness of ICU nurses.

Adaptor Proteins, Signal Transducing ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Case-Control Studies ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Membrane Proteins ; metabolism ; RNA, Messenger ; genetics