Objective To investigate and compare the killing effects of different prodrugs combined with suicide gene HSV-tk on laryngocacinoma cell, Hep-2 in vitro. Methods Retroviral expressing vector pL(tk)SN was constructed by recombinant DNA technology. Hep-2 cells were infected by the recombinant retrovirus. The positive cloning was obtained after G418 selection and were termed Hep/tk. The integration and expression of tk gene in Hep-2 cells were identified by RT-PCR and Southern blot. The growth state and prodrugs killing effect of tk gene modified cells were used to investigate the expression of tk gene and antitumour effect on Hep-2 cells. Results RT-PCR and Southern blot analysis confirmed the integration and expression of tk gene in Hep-2 cells. There was no significant difference in cell proliferation between the Hep/tk and Hep-2. After the treatment of GCV,the Hep/tk showed high sensitivity to GCV and bystander effects were observed siginificantly in vitro. However the efficiency of another two prodrugs ACV and BVdU was lower than that of GCV. Both tk-positive and tk-negative Hep-2 cells were relatively insensitive to ACV and BVdU.Treatment of tk gene modified cells mixed with different proportion parental cells shoewd obviously bystander effects.Conclusions The laryngocarcinoma cells Hep-2 have sensitive to HSV-tk/GCV system and have significant bystander effects, which might have therapeutic potential value for laryngocarcinoma.

BACKGROUND: Mesenchymal stem cells (MSCs) can differentiate into osteoblasts in the action of dexamethasone; Meanwhile, bone morphogenetic protein-2 (BMP-2) can promote the cell proliferation and differentiation, and matrix secretion in the process of bone repair. BMP-2 plays an important role in treating fracture and bone defects by inducing bone formation both in vivo and in vitro.OBJECTIVE: To analyze whether dexamethasone induction in vitro could enhance the ability of BMP-2 modified MSCs in osteogenic conversion.DESIGN: A randomized paired design.SETTING: Xijing Hospital of the Fourth Military Medical University of Chinese PLA.MATERIALS: The experiments were carried out in the Orthopaedic Oncology Institute of Chinese PLA, the Fourth Military Medical University of Chinese PLA from February to August in 2004. Twenty 20-month-old New Zealand rabbits were provided by the experimental animal center of the Fourth Military Medical University of Chinese PLA [Certificate number: 2005C00117]. The rabbits were raised normally at room temperature with normal humidity. The samples were collected bilaterally, the cells from the left limb were taken as the dexamethasone-induced group, and those from the right limb as the control group.METHODS: MSCs treated and untreated with dexamethasone were transfected with BMP-2 gene, then the BMP-2 expression was determined by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical staining. The growths of the MSCs were observed, and the activity of alkaline phosphatase (ALP) and osteocalcin of MSCs were determined with ALP kit and osteocalcin radioimmunoassay kit after Ad-BMP-2 transfection.MAIN OUTCOME MEASURES: ① BMP-2 expression in MSCs; ② Morphological changes of the MSCs; ③ ALP activity and osteocalcin content in the MSCs. RESULTS: ① Expressions of BMP-2 gene could be observed in MSCs both treated and untreated with dexamethasone after transfection. ② The forms were irregular in most of the MSCs treated with dexamethasone, appeared as triangular or polygonal changes.They grew more slowly than those cultured with basic medium. There were no obvious changes of cellular forms after gene transfection. ③ Five days after transgene, the ALP activity in the MSCs supernatant in the dexamethasone-induced group was higher than that in the control group [(134.36±8.84), (104.02±7.83) nkat/L, t =3.350 6, P ＜ 0.01, n =20]. The amount of osteocalcin secretion in MSCs in the dexamethasone-induced group was higher than that in the control group [(14.68±0.73), (6.52±1.21) μg/L, t =3.568 2, P ＜ 0.01, n =20].CONCLUSION: Dexamethasone induction before transgene can promote the proliferation of BMP-2 modified MSCs and the conversion into osteoblasts.

Purpose:To study the effect of the herbal decoction Qingyi Xiaoji Formula(QYXJ) on the proliferation of human pancreatic cancer cell line SW1990 in vivo and to explore the mechanism of its functions by means of cDNA microarray.Methods:Tumor-burdened nude mice were randomized into control group,5-FU group,and QYXJ groups at different dosages.After treatment,inhibiting rates of tumor growth were calculated.Tumor mRNA of the control group and the QYXJ group at moderate dose was extracted.The fluorescent cDNA probes were prepared,labeled with two different dyes Cy3 and Cy5,and then hybridized with cDNA microarray and scanned for fluorescent intensity.The genes with different expression were identified through the analysis of gene expression profile.Results:Inhibition rates of tumor growth in the QYXJ groups were 21.31%,38.16% and 29.09%,in the dose of 18 g/kg,36 g/kg,and 72 g/kg respectively.7 genes with reduced expression were identified,the functions of which were oncogene,protein translation and synthesis,DNA synthesis and repair,cell signal transduction,etc.Conclusions:QYXJ decoction may inhibit the proliferation of pancreatic cancer in vivo,possibly by blocking the action of an oncogene and its downstream signaling,or by regulation of protein synthesis in cancer cells.

Objective To investigate the pathologic appearances,immunohistochemical features,and genetic changes of malignant triton tumor(MTT). Methods One case of MTT was studied pathologically and immunohistochemically,and the related literatures were reviewed. Results A huge mass,demonstrated in the thorax by X ray and CT scan was seen in the posterior mediastinum in the surgery.Histologically,the tumor was composed of spindle cells with significant atypia.Some of the tumor cells had dense eosinophilic cytoplasm.Immunohistochemical staining revealed positive for myoglobin,desmin and S-100 in most of the tumor cells.The pathological diagnosis was MTT of the posterior mediastinum. Conclusion Cases of MTT in the mediastinum are very rare,with less specific clinical and imaging manifestations.The diagnosis is mainly made on the basis of pathological examination and immunohistochemical staining.

Objective To investigate the value of thyroid transcription factor-1(TTF-1) in the diagnosis and biological behavior assessment of hepatocellular carcinoma (HCC). Methods Thirty liver specimens obtained from benign lesions were analysed, among which 25 were hepatic cirrhosis and inflammatory diseases, and the other 5 were adenomas. And there were 176 specimens of liver tumors, among which 142 were HCC (well differentiated, n=12; moderately differentiated, n=57; poorly differentiated, n=73), 17 were intrahepatic cholangiocellular carcinoma (ICC) and the other 17 were liver metastatic carcinoma (MC). The expression of TTF-1 was examined immunohistochemically in the above tissues, and the difference in expression of TTF-1 among different tissues was examined by Fisher's exact test, Kruskal-Wallis test and Spearman rank correlation analysis. Results TTF-1 was significantly expressed in the cytoplasms of all the hepatocytes besides tumors and liver benign lesions. The expression rate of TTF-1 in HCC was 78.9% (112/142), however, TTF-1 was negatively expressed in ICC and MC(P

Animals ; Ginkgo biloba ; chemistry ; Lung ; blood supply ; Malondialdehyde ; metabolism ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; Superoxide Dismutase ; metabolism

Objective To develop a sheet to assess infection risks in children with malignant tumors after chemotherapies.Methods A risk assessing sheet (to consult) was designed and used to consult 24 experts from 9 hospitals located in 6 provinces.After 3 rounds of expert consultation,a sheet (to try) was developed and tried by nurses in the clinic.And then a sheet (to use) was developed.Results The authority coefficient of experts was 0.79,the positive coefficient of experts was 1.00 and 0.96.The sheet (to try) developed after 3 rounds of expert consultation consisted of 30 items and was divided into 2 parts,one was about static items and the other was about dynamic items.Depending on the trying of the sheet (to try),the sheet (to use) was constituted by 8 fundamental immune state assessing items and 16 infection risk screening items.Conclusions Experts consulted in the study were quite reliable,representative and positive.The sheet (to use) seemed to have good pertinence and practicability.Further studies should be done to verify whether the sheet (to use) may help nurses to prevent children with malignant tumors from infections after chemotherapies.

Objective To observe and compare the clinical characteristics of colorectal related multiple primary carcinoma (MPC) and colorectal carcinoma (CRC) for promoting early diagnosis and treatment of colorectal related MPC. Methods Pathological and clinical documents of clearly staged CRC and colorectal related MPC cases from Jul. 1997 to Nov. 2007 were retrospectively analyzed. Results 573 colorectal carcinoma cases were analyzed, including 45 MPC (7.85 %). Parenteral multiple carcinoma originated most frequently from stomach, and then breast, ovary, lung, small intestine and other sites. Among all multiple primary colorectal carcinomas (MPCC), ascending colon carcinoma was most frequent (34.0 %).While among CRC cases, rectal cancer cases was most frequent(36.5 %). Comparing CRC and MPCC, there were no significant difference in terms of tumor family history. Median morbidity age was 57 years and 63 years respectively. Cases with previous colonic polyps accounted for 20.0 % of all MPC cases, while only 0.9 % of all CRC cases. The mOS of CRC and MPC was 93.7 month and 64.8 month respectively. Most frequent pathological type of CRC and MPC were both well-moderately differentiated adenocarcinoma, but more mucinous adenocarcinoma cases were observed in MPC. Conclusion Colorectal related MPC are relatively common among colorectal carcinoma patients. More patients with MPC especially MPCC has colonic polyp. mOS of MPC is shorter than that of CRC, indicating the poor prognosis of MPC compared with CRC.MPCC has multiple colonic polyps, shorter interval of secondary carcinomas, and shorter mOS, worse prognosis than MPC with parenteral tumor.

Objective To intensify clinical evaluation on the manifestation, diagnosis and treatment of primary malignant duodenal tumors. Method The clinical data of 81 patients with primary malignant duodenal tumors from 1990 to 2002 were analyzed retrospectively. Result Tumors located above, around and below the duodenal papilla accounted for 16%, 78%, 6% respectively. The common clinical presentations were weight loss(72%)?abdominal pain(64%)?jaundice(45%)?alimentary tract bleeding(40%)?vomiting(33%) and anemia(24%). The preoperative diagnosis rate was 80%. Pancreatoduodenectomy was performed in 54 cases, bypass operation in 21 cases, segmental duodenectomy and simple laparotomy in 2 cases each. The postoperative 3- and 5- year survival rate was 36% and 21% for patients undergoing curative resection. In those with bypass operation, the survival time was between 1～18 months. Two cases undergoing segmental duodenectomy died within one year. Conclusion Patients usually lack special symptoms and signs, early diagnosis is fairly difficult, pancreaticoduodenectomy performed for purpose of curative resection could improve the prognosis of this malignancy.

Objective To determine the effect of transcription and translation in telomeric related proteins,and synergism of progressive telomere shortening and cell cycle alteration in human lung adenocarcinoma A549 cell line,which is induced by antisense tankyrase oligonucleotide(asTANKS) combinated with antisense human telomerase reverse transcriptase(ashTERT) oligonucleotide.Methods A549 cells were randomly assigned as 3 test groups: ashTERT,ashTERT + asTANKS and asTANKS,three control groups(shTERT,sTANKS and blank).With individual intervention for different hours,the effect of transcription in hTERT mRNA was evaluated by RT-PCR,and telomerase activity was tested by ELISA-PCR,tankyrase activity was tested by Western blot as well.Moreover,telomere average length was analyzed by Q-FISH,and duration of proliferation was observed by population double test.Results Transcription in hTERT mRNA and telomerase activity for 48 hrs was inhibited obviously by ashTERT,but not by asTANKS.Progressive telomere shortening in A549 cells for 48 hrs was induced by either asTANKS or ashTERT(vs control,P