Isolation and Biological Characterization of Lung Cancer Stem Cells / 中国医学科学院学报

To explore the methods of screening and biological characteristics of lung cancer stem cells. We selected the ABCG2 and ABCG2 cells from SPC-A-1/adriamycin(ADM)cell line induced by ADM and analyzed the tumorigenicity of ABCG2 and ABCG2 cells by flow cytometry and transplantation in nude mice. The average fluorescence intensity of SPC-A-1 cells was(1.001±0.014)×10 ,which was significantly lower than that of SPC-A-1/ADM cells [(10.257±0.023) ×10 ](=17.320,=0.001);the difference was also statistically significant between the ABCG2/BCRP-FITC treatment group and the SPC-A-1 control group(=5.269,=0.021) and the SPC-A-1 control group(=6.869, =0.012) and between the SPC-A-1/ADM cell control group and the SPC-A-1/ADM cell homotype control group(=8.112,=0.015).The positive rate of SPC-A-1/ADM cells treated with ABCG2/BCRP-FITC was 9.8%,39.84 times higher than that of SPC-A-1 cells;it showed significant difference between the ABCG2/BCRP-FITC group and the SPC-A-1/ADM group(=9.120,=0.005) and the SPC-A-1/ADM group(=8.257,= 0.006).The positive rate of group B cells was 684 times that of group A cells,and the difference was statistically significant(=11.235,=0.001),and the fluorescence intensity of group B cells was strong.The average tumorigenic volume of the mice inoculated with SPC-A-1 cells,group A cells,and group B cells was(6.96±1.82),(6.70±2.55),and(9.17±2.41) mm ,respectively.Among them,group B was the highest,but there was no significant difference among these three groups(=2.362,=0.086).The tumorigenic rate of group B cells was 75.00%,which was significantly higher than that of SPC-A-1 cells and group A cells(=19.780,=0.002). ABCG2 cells from human lung adenocarcinoma SPC-A-1/ADM cell line can be isolated by ABCG2 antibody combined with immunomagnetic beads sorting method,and the tumor formation rate in nude mice can be observed to explore the identification and biological characterization of lung cancer stem cells.

The differentiatial diagnostic value of 99Tcm-MIBI SPECT combined with localizable CT imaging in oxygen intervention for lung lesions / 中华核医学杂志

Objective To investigate the differentiatial diagnostic value of 99Tcm-methoxyisobutylisonitrile ( MIBI ) SPECT combined with localizable CT in the evaluation of oxygen intervention for suspicious lung lesions,and to establish a cost-effective imaging modality in the detection of malignant lung lesions.Methods From September 2008 to March 2009,47 consecutive patients with suspicious malignant lung lesions underwent 99Tcm-MIBI SPECT/CT prospectively. Patients with suspicious pneumonia were treated with antibiotics for about 4 d before imaging. All patients were cannulized with a nostril tube for oxygen inhalation before 99Tc-MIBI injection. SPECT combined with localizable CT of the chest was performed at 10 min and 2 h after 99Tcm-MIBI injection. The uptake ratios of lesion to contralateral normal lung parenchyma(early uptake ratio:EUR and delayed uptake ratio:DUR) were compared using independent-samples ttest. In addition,the diagnostic efficiency of uptake ratios of lung lesions was analyzed with receiver operating characteristic (ROC) curve. Results Forty-seven patients ( primary lung cancers:32,metastatic tumors of the lung:4,benign lung diseases:11 ) had 51 lung lesions,including 39 malignant and 12 benign lung lesions. The sensitivity,specificity,accuracy,positive predictive value (PPV),and negative predictive value (NPV) were 94.9% (37/39),83.3% ( 10/12),92.2% (47/51),94.9% (37/39) and 83.3% ( 10/12),respectively. The EUR of malignant lesions was 2.95 ± 1.16,whereas of benign lesions was 1.43 ±0.33. The DUR of the malignant lesions was 3.19 ± 1.74,whereas of benign lesions was 1.60 ±0.32. Both EUR and DUR were significantly different between malignant and benign lung lesions,respectively (t= -4.44,-3.12,respectively,both P＜0.01). The ROC curve showed that EUR value ≥1.625 provided the sensitivity of 97.4% (38/39) and specificity of 83.3% (10/12); DUR value ≥ 1.75provided the sensitivity of 94.9% (37/39) and specificity of 83.3% (10/12). Conclusion 99Tcm-MIBI SPECT combined with localizable CT imaging in oxygen intervention has a potential value in differentiating malignant from benign lung lesions.

Cloning, Sequencing of Suaeda heteroptera kitag CMO cDNA and Construction of its Recombinant Plant Expression Vector / 中国生物工程杂志

Total RNA was extracted from leaf of Suaeda hetroptera kitag, then the CMO ( choline monooxygenase) cDNA was amplified using the reverse transcriptase polymerase chain reaction ( RT-PCR) method and cloned into pMD-T-simple vector. The positive clones from the Blue/White Screen were sequenced. After confirming its validity, the CMO gene fragment was cloned into pBI121 vector. Double enzyme restriction and PCR analysis indicated that the pBI121/CMO recombinant plasmid was successfully constructed.

Expressions and activities of Rho GTPases in hypoxia and its relationship with tumor angiogenesis / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the expressions and activities of Rho GTPases in hypoxia and its relationship with tumor angiogenesis.</p><p><b>METHODS</b>Three tumor cell lines were used in this study: gastric cancer cell lines AGS, SGC7901 and hepatocellular carcinoma cell line HepG2. Expression level of Rac1 mRNA was detected by semi-quantitative RT-PCR. Activity of Rac1 was determined by pull-down assay and expression of HIF-1alpha, VEGF, p53 and PTEN protein was detected by Westernblot.</p><p><b>RESULTS</b>The expression level of Rac1 mRNA was significantly increased in hypoxia compared to normoxia. Pull-down assay showed that hypoxia-induced activity of Rac1 was elevated in a time-dependent manner and climaxed at 3 hours. The expressions of HIF-1alpha and VEGF protein were up-regulated, while those of PTEN and p53 protein were down-regulated.</p><p><b>CONCLUSION</b>These results indicate that hypoxia enhances Rac1 expression which might be involved in tumor angiogenesis by reacting with hypoxia-responsive genes.</p>

Differential expression of RPL6/Taxreb107 in drug resistant gastric cancer cell line SGC7901/ADR and its correlation with multiple-drug resistance / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the differential expression of RPL6/Taxreb107 between drug-resistant gastric cancer cell line SGC7901/ADR and gastric cancer cell line SGC7901 as well as its correlation with multiple-drug resistance (MDR) in gastric cancer cells.</p><p><b>METHODS</b>Total RNA was extracted from SGC7901 and SGC77901/ADR, with internal control RT-PCR, Northern blot, gene cloning and expression, construction of eukaryotic expression vector, gene transfection by electroporation. The accumulation and retention of ADR in transiently transfected cell was detected by flow cytometry.</p><p><b>RESULTS</b>The internal control RT-PCR and Northern blot showed high RPL6/Taxreb107 expression in SGC7901/ADR cell line. Sense and antisense eukaryonic expression vectors demonstrated by double enzyme digestion were successfully transfected into gastric cancer cell line SGC7901 and SGC7901/ADR respectively by electroporation. The accumulation and retention of ADR detected 48 hours after transfection showed that RPL6 gene had shown effect on drug resistance in gastric cancer cell.</p><p><b>CONCLUSION</b>The high expression of RPL6/Taxreb107 in drug resistant gastric cancer cell shows its correlation with multiple-drug resistance in gastric cancer.</p>

Fermentative technology of Soybean Isofiavone Glucoside Hydrolase-Producing Strain / 微生物学通报

A high active soybean isoflavone glucoside hydrolase-producing mould strain was isolated from spirit qu. Its optimal hydrolase-producing conditions were as follows: 2.5% wheat bran as carbon source, 1% NaNO3 as nitrogen source, initial pH7. 0, culture medium volume 40mL/250mL, inoculating quantity 8% , culture temperature 30℃, revolutions 160r/min and culture time 84h. The enzyme activity reached 82 U/mL. Cu2+ can inhibit Absidia sp. R strain from producing the hydrolase, the influence of other metal ions was not remarkable on it.

STUDY ON FERMENTATION CONDITIONS OF A 1.3-PD HIGH-PRODUCTION STRAIN FROM E. AEROGENES / 微生物学通报

The fermentation conditions of high 1.3 -propanediol-producing strain E. aero-N-56 were determined in this Paper. The optimum conditions of producing 1.3-PD were: initial pH 7.0, temperature 30℃, culture time 48 h, inoculum size 9% . Under the optimum conditions: the 1.3-PD productivity reached up to 23.68 g/L?d; the 1,3-PD yield of E. aero-N-56 up to 47.36 g/L in 30 L fermentor.