Objective To analyze non -small cell lung cancer patients with platinum -based chemotherapy excision repair cross -complementing 1 (ERCC1)and breast cancer susceptibility gene 1 (BRCA1 )gene polymor-phism,to examine the correlation of ERCC1 and BRCA1 gene polymorphism and non -small cell lung cancer platinum chemotherapy drugs sensitivity and chemotherapy prognosis.Methods 140 cases of non -small cell lung cancer were selected as subjects of this study.All patients were given platnum -based chemotherapy,peripheral blood ERCC1 and BRCA1 genes polymorphism were determined.The distribution of ERCC1 Asn118Asn genotype and BRCA1 Ser1613Gly genotype was observed.The relationship between different genotypes and the effect of chemotherapy and survival time after chemotherapy was compared.Results The proportions of ERCC1 Asn118Asn TT genotype,CT genotype and CC genotype were 5.7%,30.7% and 63.6%.The proportions of BRCA1 Ser1613Gly GG genotype,AG genotype and AA genotype were 8.6%,52.9% and 38.6%.In 140 patients,completely cured,partial response,stable disease and progressive disease patients were 0 case,33 cases,61 cases and 46 cases,the proportions were 0.0%, 23.6%,43.6% and 32.9%,the chemotherapy effective rate was 33.8%.ERCC1 Asn118Asn genotype was signifi-cantly correlated with the effect of non -small cell lung cancer chemotherapy (χ2 =4.416,P <0.05 ).BRCA1 Ser1613Gly genotype was significantly correlated with the effect of non -small cell lung cancer chemotherapy (χ2 =13.256,P <0.05).By Cox regression analysis and Log -rank test analysis,the average survival time of BRCA1 Ser1613Gly gene CC genotype non -small cell lung cancer after chemotherapy was longer than the CT +TT genotype (OR =2.946,χ2 =5.136,P <0.05).The average survival time of BRCA1 Ser1613Gly gene AA genotype non -small cell lung cancer after chemotherapy was shorter than the AG +GG genotype (OR =3.124,χ2 =5.136,P <0.05).Conclusion ERCC1 Asn118Asn genotype and BRCA1 Ser1613Gly genotype was significantly correlated with non -small cell lung cancer platinum -based chemosensitivity and chemotherapy prognosis.

Objective:To construct double copy and x gene deleted HBV expression plasmid and study its expression in Hep3B cell line. Methods:The double copy HBV DNA ( adr Ⅰ) was used to inactivate HBV x gene by inserting mutation and gene recombination. The inserted 55 bp DNA sequence was synthesized artificially; the insertion point was ApaL Ⅰ of x gene area. After recombination, an x gene defected HBV plasmid containing single P, S and C gene was constructed,which can express in mammalian cell line. Another plasmid carrying double copy HBV DNA with normal x gene was constructed as contrast. Both were used to transfect Hep3B cells. Then the cells were screened by G418 and HBV virus in culture medium were isolated and detected by fluorescence quantitative PCR. Results: Plasmids pcDNA3 KN F1F2 and pcDNA3 ES HBV2 were constructed successfully. After cell transfection, the HBV DNA was highly expressed with both plasmids on the 3 rd, 6 th,14th day. Conclusion: The plasmids constructed can express in Hep3B cell line and cause HBV replication; x gene defected HBV gene has no effect on HBV replication in Hep3B cell line.

OBJECTIVETo study the injection of NKG5SV gene to inhibit growth and metastasis of hepatocellular carcinoma (HCC).

METHODSNKG5SV gene was inserted into retroviral vector pLXSN by normal methods. LacZ gene was used as control. LCI-D20 tumor together with saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was subcutaneously inoculated to the nude mice. Tumor formation rate and tumor size were noted 35 days after inoculation. LCI-D20 tumor was inoculated subcutaneously. Saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was intratumorally injected respectively 10 days after inoculation. Tumor growth was observed 35 days after inoculation. Liver cancer was resected 22 days after intrahepatic inoculation. Saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was respectively injected at incisal margin or intraspleen. Mice were killed 35 days after inoculation to observe tumor recurrence at incisal margin, intrahepatic metastasis and extrahepatic metastasis.

RESULTSTumor formation rate and tumor diameter(cm) were 1.76 +/- 0.11, 1.51 +/- 0.34, 0.33 +/- 0.04 in the control group, LacZ group, NKG5SV group respectively when tumor and different cDNA were inoculated together. Tumor diameter(cm) and weight(g) were 0.87 +/- 0.08, 0.83 +/- 0.05, 0.26 +/- 0.04; 0.43 +/- 0.06, 0.38 +/- 0.04, 0.08 +/- 0.06 in the control group, LacZ group, NKG5SV group respectively when different cDNA were injected into the LCI-D20 tumor. Sites with extrahepatic metastasis nidi, incisal margin recurrence tumor size(cm), intrahepatic metastasis nidi, metastasis involved hepatic lobes in the control group, LacZ group, NKG5SV group were 4.25 +/- 1.48, 4.25 +/- 1.04, 0.63 +/- 0.51; 1.51 +/- 0.27, 1.35 +/- 0.17, 0.81 +/- 0.17; 2.50 +/- 1.41, 2.38 +/- 1.06, 1.25 +/- 0.71; 2.13 +/- 0.99, 2.00 +/- 0.75, 1.38 +/- 0.74 respectively when NK cells were injected at incise margin. They were 4.38 +/- 1.85, 4.25 +/- 1.48, 1.00 +/- 0.75; 1.13 +/- 0.23, 0.97 +/- 0.29, 0.76 +/- 0.16; 2.50 +/- 1.41, 2.05 +/- 1.12, 0; 2.13 +/- 0.83, 1.75 +/- 0.88, 0 respectively when NK cell were injected intrasplenicly.

CONCLUSIONSNKG5SV gene can inhibit HCC growth and postoperative metastasis and recurrence.

Animals ; Antigens, Differentiation, T-Lymphocyte ; Cell Division ; drug effects ; Genetic Therapy ; methods ; Genetic Vectors ; administration & dosage ; genetics ; Humans ; Injections ; Liver Neoplasms, Experimental ; genetics ; pathology ; therapy ; Male ; Mice ; Mice, Nude ; Neoplasm Metastasis ; prevention & control ; Receptors, Immunologic ; genetics ; physiology ; Tumor Cells, Cultured ; Xenograft Model Antitumor Assays

OBJECTIVETo develop a new kind of vector system called gene-viral vector, which combines the advantages of gene and virus therapies.

METHODSUsing recombinant technology, an anti-tumor gene was inserted into the genome of replicative virus specific for tumor cells. The cell killing effect, reporter gene expression of the green fluorescence protein, anti-tumor gene expression of mouse interleukin-12 (mIL-12) and replication of virus were observed by the methods of cell pathology, fluorescence microscopy, ELISA and electron microscopy, respectively.

RESULTSA new kind of gene-viral vector system of adenovirus, in which the E1b-55 kD gene was deleted but the E1a gene was preserved, was constructed. The vector system, like the replicative virus ONYX-015, replicated and proliferated in tumor cells but not in normal ones. Our vector had an advantage over ONYX-015 in that it carried different kinds of anti-tumor genes to enhance its therapeutic effect. The reporter gene expression of the green fluorescence protein in tumor cells was much better than the adenovirus vector employed in conventional gene the rapy, and the expression in our vector system was as low as or even less than that in the conventional adenovirus gene therapy system. Similar results were observed in experiments with this vector system carrying the anti-tumor gene mIL-12. Replication and proliferation of the virus carrying the mIL-12 gene in tumor cells were confirmed by electron microscopy.

CONCLUSIONSGene-viral vectors are new vectors with an anti-tumor gene inserted into the genome of replicative virus specific for tumor cells. Because of the specific replication and proliferation of the virus in tumor cells, expression of the anti-tumor gene is increased hundreds to thousands of times. This approach takes full advantages of gene therapy and virus therapy to enhance the effect on the tumor. It overcomes the disadvantages of conventional gene therapy, such as low transfer rate, low gene expression, lack of target tropism, and low anti-tumor activity. We believe that this is a promising means for future tumor treatment.

Adenoviridae ; genetics ; Adenovirus E1A Proteins ; genetics ; Adenovirus E1B Proteins ; genetics ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Humans ; Interleukin-12 ; genetics ; Neoplasms ; therapy ; Recombination, Genetic ; Tumor Cells, Cultured ; Virus Replication

Objective:To construct 68Ga-1, 4, 7-trizacyclononane-1, 4, 7-triacetic acid (NOTA)-CD44 as a novel atherosclerosis tracer targeting hyaluronic acid (HA), and evaluate its biological property and molecular imaging features. Methods:Low molecular weight (LMW) recombinant human CD44 protein was selected, and the C-terminal of the protein was modified by sulfonation and coupled to the bifunctional ligand NOTA to synthesize a novel molecular probe 68Ga-NOTA-CD44 targeting HA. The biological properties of the probe, such as labeling rate and in vitro stability, were studied. Three atherosclerotic plaque model mice and three normal C57BL/6 mice were studied by 68Ga-NOTA-CD44 microPET/CT imaging and pathological examination. Results:68Ga-NOTA-CD44 tracer was synthesized and purified with the radiochemical purity above 99%, and the specific activity was up to 62.22 MBq/nmol. lts stability was good in PBS, and the radiochemical purity was over 90% after incubation for 3 h. After intravenous injection, the probe was metabolized mainly by the kidneys, and its metabolic level decreased successively in the liver, lungs and blood. MicroPET/CT imaging results of atherosclerotic model mice suggested that the uptake in the plaque of abdominal aorta was higher at 60 min after injection, with SUV max and target/background ratio (TBR) max of 1.14±0.02 and 4.95±0.93, and the probe had certain atherosclerotic plaque eroded targeting, which was consistent with the pathological result. Conclusions:As a novel probe, 68Ga-NOTA-CD44 is simple to prepare and has a high labeling rate. It has good physicochemical properties and in vivo biological properties, and can display atherosclerotic eroded plaques sensitively. 68Ga-NOTA-CD44 has a promising prospect to be a new molecular probe for early noninvasive recognition of atherosclerotic eroded plaques.