Objective: To determine if fusion genes of HSV-tk gene and cytokine gene have synergy on the cell killing of the Hep-2 human laryngeal carcinoma cell line in vitro. Methods: Different fusion genes expressing vectors PL(TI) SN, PL(TT)SN and PL(TK)SN were generated by recombinant DNA technology. Hep-2 was infected by the recombinant retrovirus. The positive clones were obtained after G418 selection and were termed Hep/TI, Hep/TT and Hep/TK respectively. The integration and expression of fusion genes in Hep-2 cells were identified by RT-PCR and Southern blot. The growth state and GCV killing effect of fusion genes modified cells were used to investigate the expression of fusion genes and antitumour effect on Hep-2 cells. Results: RT-PCR and Southern blot analysis confirmed the integration and expression of fusion genes in Hep-2 cells. There was no significant difference in cell proliferation between the Hep/TI and Hep/ TK.but the growth of Hep/TT was restrained. After the treatment of GCV the Hep/TI, Hep/TT and Hep/TK all showed high sensitivity to GCV. The killing effect of GCV on Hep/TT was the most siginificant and bystander effects were observed siginificantly in vitro. Conclusion: The fusion genes of HSV-tk and cytokine gene have synergistic effects on killing Hep-2 cell after treatment of GCV in vitro,which might have therapeutic potentials for laryngocarcinoma.

Objective To explore the diagnostic significance of C reactive protein(CRP)in diagnosing haemorrhagic fever with renal syndrome(HFRS).Methods 96 cases of patients with HFRS of different stages were enrolled in this study,and serum speci-men were collected.30 cases of patients with fever of unknown origin(fever of unknown origin group)and and 30 healthy individu-als(healthy control)were selected as control.Serum levels of CRP,alanine aminotransferase(ALT),aspartate amino-transferase (AST),creatine phosphokinase(CK),lactate dehydrogenase(LDH)and creatinine(Cr)were detected.Results Serum levels of CRP in patients with HFRS of different stages were lower than that in the fever of unknown origin group,had statistically significant differences(P <0.05).The variation trend of CRP in each stage of HFRS was consistent with the trend of AST,CK,LDH and Cr. Conclusion CRP has clinical significance in differentiating HFRS from fever of unknown origin.

Adenosine deaminase (ADA)plays an important role in the regulation of normal immune system.The altered activ-ity of serum ADA could associate with some autoimmune diseases(AID).Various studies abroad have found the elevated ac-tivity of serum ADA compared with healthy controls in many kinds of AID such as systemic lupus erythematosus (SLE), rheumatoid arthritis(RA),et al (P<0.05).Many studies foused on the correlation of altered activity of serum ADA with the disease severity,but there is still controversy.To explore the diagnostic value of serum ADA in AID,make a review a-bout the application research progress of ADA in several AID.

Objective To analyze the results of distal tibia and fibula fracture treated with delayed open reduction and internal fixation with anatomic plate.Methods 24 fracture of distal tibia and fibula were stabilized temporanrily by application of plaster splint or calcaned traction immediately after injury.The definitive internal fixations were performed on an average of 8 days(range 7 to 10 days) after injury as soon as the soft tissue recovered.Results All patients were available for follow-up at an average of 2.6 years(range,1 to 4.5 years) after surgery.All fracture headed at an average of 15.4 weeks(range,12 to 29 weeks) postoperatively.There were 16 excellent,6 good,1 fair,and 1 poor results.The excellent and good rate was 92%.Conclusion This two-stage treatment protocol for fracture of distal tibia and fibula leads to the best long-term outcomes.

Assay"'of Epstein-Barr virus (EBV) DNA was conducted in the tissues of 60 patients with nasopharyngeal carcinoma (NPC), 30 patients with chronic inflamation of nasopharyngeal mucous membrane and 40 patients with other malignant epithelial tumor respectively using DNA in situ hybridization method. The positive rate of EBV-DNA in patients of NPC was 71.6%, while it was negative in pat ents with other malignant tumors a small number of EB-VDNA positive cells were also discovered in the epithelial cells of paratumours and chronic inflamation of nasopharyngeal mucous membrane. Significant correlation was found in the study of serum EBV-antiby in patients of NPC and EBV-DNA in cancer cells. The results suggest that EBV may be of etiologic significance in the pathogenesis of NPC.

Purpose:To investigate the expression and significance of VEGF in lung carcinoma.Methods:SP immu- nohistochemical staining was used to detect the expression of VEGF and MVD in 42 cases of lung carcinoma.Results:The expression rate of VEGF was 81.0% in lung carcinoma.In the group of VEGF positive staining,microvessel density (MVD) was (45.68?9.23) per high power field which was significantly larger than(34.58?10.22) per high power field in the group of VEGF negative staining (P

Objective To learn the exposure levels and health hazards of long-term exposure to hexavalent chromium (Cr6+) through drinking water in a west county,and to provide a scientific basis for making preventive measures.Methods Five water points were selected to test the Cr6+ concentration of drinking water in the county in 2015,data of 3 water points with water Cr6+ concentrations exceeded the standard (Cr6+ ＞ 0.05 mg/L) were selected from 12 drinking water points in some west counties in the recent six years (2009-2014) as the exposed group,2water points that drinking water Cr6+ concentrations not exceeded the standard (Cr6+ ≤0.05 mg/L) in the county and adjacent to the exposed group were selected as the control group.Sixty villagers were selected as the investigation objects in each water point to conduct internal medicine,ears,nose,throat (ENT),dermatology and health examination,urinary chromium content,routine blood and urine test were done.Determination of hexavalent chromium concentration in drinking water was done according to The Drinking Water Standard Examination Method 1,5-diphenylcarbazide Spectrophotometry (GB/T 5750-2006);routine urine was tested using the 10 urine analyzer test;urinary chromium was tested using graphite furnace atomic absorption spectrometer;routine blood five classification was tested using automatic blood analyzer;determination of drinking water hexavalent chromium concentration was done according to The Hygienic Standard for Drinking Water (GB 5749-2006).Higher than 0.05 mg/L was judged as exceeded the standard;physical examination was done according to The Diagnostic Criteria of Occupational Chromium Nasal Disease (GB Z12-2002) and The Diagnostic Standard of Occupational Contact Dermatitis (GB Z20-2002).Results There were 184 and 109 people in the exposed group and the control group,respectively.The average concentration of Cr6+ exceeding the standard ratio was 2.82-3.22 in drinking water,the nasal septum nucosa perforation,skin erythema edema,skin ulcers were 4.9％ (9/184),4.3％ (8/184) and 4.3％ (8/184) in the exposed group,and the urinary chromium level (0.31 μg/L) in the exposed group was significantly higher than that of the control group (0.27 μg/L,Z =-4.078,P ＜ 0.05).Routine blood test result of mean corpuscular haemaglobin (MCH) was (29.56 ± 2.07) pg,platelet counts (PLT) was (222.38 ± 47.53) × 109/L and plateletcrit (PCT) was (0.25 ± 0.05)％ in the exposed group,it was all higher than those of the control group [(29.03 ±2.95) pg,(211.74 ±75.27)× 109/L,(0.24 ± 0.08)％,t =1,940,2.318,2.079,all P ＜ 0.05];standard difference coefficient of variation of red blood cell distribution width (RDW-CV) was (13.14 ± 1.05)％,lymphocyte number (LYMPH) was (2.01 ± 0.64) × 109/L in the exposed group,it was all lower than those of the control group [(13.38 ±1.54)％,(2.21 ± 1.02) × 109/L,t =-1.989,-1.956,all P ＜ 0.05].The positive rate of glycosuria,uric bravery red,and urine nitrite in routine urine test of exposed group was 17.39％ (32/184),23.36％ (43/184),7.61％ (14/184);it was all higher than those of the control group [(8.25％ (9/109),11.93％ (13/109),0.91％ (1/109),x2 =4.746,5.798,6.309,all P ＜ 0.05].Conclusions Urine chromium accumulation has been found in populations long-term exposed to 2.82-3.22 times excessive Cr6+ through drinking water,which has affected the population's health to some extent.Therefore,it is necessary to speed up the local drinking water improvement project.

Purpose　To observe the killing effect of HSV-TK/GCV system on osteosarcoma cells. Methods　The eukaryotic exprssoin vector (tgCMV-HyTK) containing HSV-TK gene was transduced into human osteosarcoma cell lines SAOS-2 by DOTAP. Total RNA extracted from HSV-TK expressing cells (SAOS-2TK) were tested by RNA dot blot analysis. A chemosensitivity of SAOS-2TK cells to GCV and “bystander effect” were measured by means of MTT colrimetric assay. Results　HSV-TK gene was expressed in SAOS-2TK cells. SAOS-2TK cells were susceptible to the cytotoxicity of GCV. A meaningful “bystander effect” was demonstrated. Conclusion　HSV-TK/GCV system may kill selectively the human osteosarcoma cell line SAOS-2.

Objective: To construct the eukaryotic expression vector pSurp-EGFP regulated by the survivin gene promoter and to detect the specific expression of the promoter in human laryngeal cancer Hep-2 cells by green fluorescent protein assay.Methods: Thesurvivin gene promoter was generated by polymerase chain reaction(PCR) and the CMV promoter of the pShuttle vector replaced by the survivin gene promoter to generate the plasmid pSurp.The three plasmids pShuttle,pSurp and pEGFP-C1 were respectively double-enzyme digested so as to produce the plasmids pCMV-EGFP and pSurp-EGFP carrying the CMV or survivin promoter.The purified pCMV-EGFP and pSurp-EGFP were transfected into Hep-2 cell and vascular endothelial cell ECV304 using liposome transfection reagent and the expressions of EGFP detected by the fluorescent microscope.Results: Thesurvivin gene promoter was successfully cloned by PCR,and thesurvivin gene promoter-regulated pSurp-EGFP was constructed.Green fluorescence was observed in Hep-2 cells but not in ECV304. Conclusion: The high specific activity of the survivin gene promoter in Hep-2 cells that we successfully constructed attributes to the studies of tumor specific gene therapy.

The incidence of rectal gastrointestinal stromal tumor is low,adequate diaganosis depends on the histopathological and immunohistochemical examination.Its treatment is a comprehensive therapy including surgery and molecular targeted therapy etal.The rectal gastrointestinal stromal is easily recurrent after operation,and so needs surveillance and following up.