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25 December 2011, Volume 3 Issue 4 Previous Issue    Next Issue

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Construction of the lentiviral expression system for small proteins YIN Qiao-Yun, LI Li, YU Hong-Jing, WANG Qi 2011, 3 (4):  271-276.  doi: 10.3969/j.issn.1674-5671.2011.04.01 Abstract ( 742 )   PDF (2719KB) ( 2513 )   Save Objective For further study the function and mechanism of chemotactic protein，to establish a lentiviral expression system and to construct a human ovarian cancer cell line that can express green fluorescent protein (GFP) stably.Methods Chemotactic factor CCL18，IL-8 and CXCL1 gene plasmids were amplified byPCR,restricted by endonuclease digestion and connected with lentiviral vector PWPI.The recombinant lentiviral vector was then mixed with the envelope plasmid PCMV-dR8.74 and packaging plasmids PMD2.G in proportion.LipofectinTM2000 293T cells were co-transfection with packaging viral particles.The viruses harvested from the supernatant were used to transfect ovarian cancer SKOV3 cells.The cells that strongly expressed GFP were selected by FACS，which served as the successfully transfected cells.The target genes were measured by Real-time fluorescent quantitative PCR. Western blot was adopted to confirm the expression levels of the target proteins.Results The efficiency of infection of the cells that express target gene was more than 95%.Real-time fluorescent quantitative PCR tests showed that the expression level of target gene improved significantly comparing to the control cells and tissues (P＜0.05). The expression of chemotactic protein in the transfected cells was validated by Western blot. Conclusion A lentiviral expression system for small protein is successfully constructed，which is an experimental basis for further study the function and mechanism of small-molecule protein. Related Articles | Metrics

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Correlation between IGF-1R polymorphism and genetic susceptibility to hepatocellular carcinoma QIN Yan-Xia, HUANG Tian-Ren, DENG Wei, YU Jia-Hua, ZHANG Chun-Yan, LI Ji-Lin 2011, 3 (4):  277-280.  doi: 10.3969/j.issn.1674-5671.2011.04.02 Abstract ( 548 )   PDF (806KB) ( 769 )   Save Objective To explore the correlation between IGF-1R polymorphism and genetic susceptibility to hepatocellular carcinoma (HCC) in the area with high incidence of HCC in Guangxi.Methods A case-control study was conducted which included 113 HCC patients and 113 normal controls.All of the cases were from the areas with high incidence of HCC in Guangxi.The SNP locus rs 61740868 of gene IGF-1R was selected as a genetic marker，and its genotypes as well as mutation frequency were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).Results There was no mutation in SNP locus rs 61740868 of IGF-1R in both HCC cases and controls.The expressions of IGF-1R were not statistically different between the two groups (P＞0.05).Conclusion Polymorphism of IGF-1R SNP locus rs 61740868 is not associated with HCC，it may not be a risk factor for HCC. Related Articles | Metrics

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The effects of chloroquine diphosphate and rapamycin at different concentration on autophagy of CNE-2 cells PAN Wen-Yan, ZHU Xiao-Dong, ZHAO Wei, QU Song, LI Ling, SU Fang, LI Xiao-Yu 2011, 3 (4):  280-283.  doi: 10.3969/j.issn.1674-5671.2011.04.03 Abstract ( 1116 )   PDF (873KB) ( 2143 )   Save Objective To investigate the effects of chloroquine diphosphate (CDP) and rapamycin at different concentration on autophagy of CNE-2 cells. Methods Western-blot were used to detect the expression level of autophagy-related protein LC3-II (microtubule-associated protein 1 light chain 3，MAP1-LC3-II) and p62 in nasopharyngeal carcinoma cell line CNE-2 that were treated with CDP and rapamycin at different concentrations.Propidium iodide (PI) staining was used to detect the apoptosis rate of CNE-2 cells.Results Among the CDP-treated groups，the expression levels of LC3-II and p62 increased more pronouncedly when CNE-2 cells were treated at the concentration of 20.60mg/L (P＜0.01).Among the rapamycin-treated groups，18.28μg/L group showed increased expression of LC3-II (P＜0.01) and reduced expression of p62 (P＜0.01).The apoptosis rates in 20.60mg/L CDP group and 18.28μg/L rapamycin group were not statistically different from that of the control group (P＞0.05).Conclusions Chloroquine diphosphate can inhibit autophagy of CNE-2 cells at the concentration of 20.60mg/L.Rapamycin can induce autophagy at the concentration of 18.28μg/L.Both of them do not affect the apoptosis rate of CNE-2 cells. Related Articles | Metrics

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The relationship between the expressions Stat3 and HIF-1α in Hep-2 cells under hypoxia LU Xiu-Ying, LI Xiao-Ming 2011, 3 (4):  284-287.  doi: 10.3969/j.issn.1674-5671.2011.04.04 Abstract ( 521 )   PDF (937KB) ( 502 )   Save Objective To investigate the expressions of p-Stat3，HIF-1α and VEGF in Hep-2 cells under hypoxia，and the effect of Stat3 antisense oligonucleotides (AS-ODN) on these expressions.Methods Expressions of p-Stat3，HIF-1α and VEGF of Hep-2 cells that were with or without Stat3 AS-ODN transfection，at hypoxic and normoxic conditions，were detected by flow cytometry.The survival rate of Hep-2 cells transfected with Stat3 AS-ODN and control groups were analyzed by MTT assay.Results The FI of p-Stat3 was up-regulated in 3h-hypoxia group，and it was stable after 6h at hypoxic condition.The FI of HIF-1α and VEGF increased after 6h and 12h at hypoxic condition. There was positive correlation between the expression of p-Stat3 and HIF-1α (r=0.691，P＜0.01).Compared to Stat3 S-ODN group and   lipofectamine  control group，the expressions of p-Stat3，HIF-1α and VEGF in Stat3 AS-ODN group at hypoxic condition were significantly decreased (P＜0.01 ). Pearson correlation analysis showed the significant positive correlation between the expressions of p-Stat3 and HIF-1α (r=0.912，P＜0.01).The survival rate of Hep-2 cell   reduced with the concentration augmentation of Stat3 AS-ODN (100nmol/L and 200nmol/L)，which were significantly different from the Stat3 S-ODN group and  HIF- 1α  and VEGFlipofectamine control group (P＜0.01).Conclusions Hypoxia can induce Stat3 activation in Hep-2 cells.The expression of p-Stat3 is earlier than that of  HIF-1α and VEGF.Blocking Stat3 can decrease the expression of HIF-1α under hypoxia，indicating Stat3 might modulate HIF-1α expression in Hep-2 cells at the hypoxic condition. Related Articles | Metrics

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Gene polymorphisms of GSTM1 and GSTT1 in the clustering families of hepatocellular carcinoma XIAO Kai-Yin, LI Le-Qun, PENG Min-Hao, QIN Xiao, PENG Tao, GUO Ya, CHEN Bin, LU Jing-Ning, QIN Quan-Lin, GUI Wen-Bo 2011, 3 (4):  287-290.  doi: 10.3969/j.issn.1674-5671.2011.04.05 Abstract ( 598 )   PDF (719KB) ( 683 )   Save Objective To investigate the relationship between gene polymorphisms of glutathione-S-transferase M1 (GSTM1) and T1 (GSTT1) and hepatocellular carcinoma (HCC)-clustering family.Methods  the genotype of GSTM1 and GSTT1 in the HCC-clustering family and high-incidence pedigrees of HCC were detected with PCR.Results The frequencies of GSTM1 null and GSTT1 null genotype were 68.8% and 47.5% in the group of HCC-clustering family respectively, which were significantly higher than those of the none-clustering HCC family (54.6% and 30.8%) and control group (53.3% and 25.3%).The same tendency was also found between highincidence pedigrees of HCC and control pedigrees. With the increasing number of HCC patient in familial history, the frequencies of GSTM1 null and GSTT1 null genotype increased too.The risk of HCC increased with the GSTM1 null genotype combined GSTT1 null genotype. Conclusion  Gene polymorphisms of GSTM1 and GSTT1 are associated with the susceptibility of HCC-clustering family. GSTM1 null genotype and GSTT1 null genotype are the risk factors in HCC-clustering family. Related Articles | Metrics

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Effect of fentanyl on growth and apoptosis of human hepatoma cell line bel-7404 ZOU Chun-Yun, RUAN Lin, ZHANG Yuan 2011, 3 (4):  291-294.  doi: 10.3969/j.issn.1674-5671.2011.04.06 Abstract ( 517 )   PDF (945KB) ( 1744 )   Save Objectives To study the effects of fentanyl on growth and apoptosis of human hepatoma cell line bel-7404.Methods bel-7404 cells were divided into four test groups ( F1，F2，F3 and F4) and a control group (C).Cells in the four test groups were incubated in RPMI-1640 medium that contained different concentration of fentanyl respectively，such as 5，50，500 and 5000 ng/ml. Cells in the control group were cultured in RPMI-1640 medium only. After 24-hour incubation，morphological changes of the cells were observed by inverted microscope，the level of cell proliferation were evaluated by the methods of MTT and colony formation，and thedistribution of cell cycle and the apoptosis rate were detected with flow cytometry (FCM). Results The results from MTT and colony formation on the test groups were significantly lower than that of the control group(P＜0.01). The percentage of apoptosis was significantly higher in the test groups than that of the control group(P＜0.05).When the concentration of fentanyl was ≥50ng/ml，the rate of apoptotic cells increased gradually with the increasing concentration of fentanyl，and the ratio of cells in G0/G1 phase enhanced gradually and in S phase decreased gradually，which was significantly different from that of the control group (P＜0.05). Conclusion The inhibitory effect of fentanyl on growth of bel-7404 cell is dose-dependent. Fentanyl is able to block cell cycle and to induce apoptosis. Related Articles | Metrics

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Inhibitory effect of norcantharidin on bone metastasis in nude mice model of NCI-H446 GAO Zhi-Di, PAN Qi, GAO Wei, MA Xiao-Ye, ZHU Feng-Qing 2011, 3 (4):  294-298.  doi: 10.3969/j.issn.1674-5671.2011.04.07 Abstract ( 521 )   PDF (731KB) ( 554 )   Save Objective To investigate the effect of norcantharidin on bone metastasis of human small cell lung cancer(SCLC). Methods A skeletal metastasis model of SCLC was established by injecting NCI-H446 cells into the right leg of nude mice. Sixty male health BALB/C mice were divided randomly into 3 groups including control group，high-dose group and low-dose group. The effects of norcantharidin on bone metastasis and bone metabolism were investigated by X-ray，pathological and   molecular biochemical detections，while the conditions of tumor growth were observed. Results Comparing to drug groups，the control group were with faster tumor growth，larger tumor size，and lower total weight gain at the end of the experiment. The levels of serum PICP and PINP were higher in control group，but which were not statistically different from that of the drug groups. Conclusions Norcantharidin can inhibit bone metastasis of lung cancer. Serum PICP and PINP levels can be used for early diagnosis and treatment monitoring of bone metastasis of SCLC. Related Articles | Metrics

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Values of diffusion-weighted imaging in differential diagnosing necrosis and recurrence of nasopharyngeal carcinoma after radiotherapy YANG Bai-Hua, CHEN Ying, XU Lu-Ying, YAO Yi-Qi, WU Jun-Xin 2011, 3 (4):  298-302.  doi: 10.3969/j.issn.1674-5671.2011.04.08 Abstract ( 660 )   PDF (997KB) ( 911 )   Save Objective To evaluate the role of diffusion-weighted imaging (DWI) and the value of apparent diffusion coefficient (ADC) in differential diagnosing necrosis and  recurrence of nasopharyngeal carcinoma after radiotherapy.Methods From Jul 2008 to Dec 2010，thirty-five patients were diagnosed postradiation nasopharyngeal necrosis，which were matched with the same number of patients randomly selected from the same period. All patients were confirmed pathologically，and received routine MRI and DWI (with the b values of 0s/mm2 and 800s/mm2). ADC values of necrotic foci and parts of recurrence tumor were measured，and their mean ADC values were calculated.Results  Nasopharyngeal necrosis usually showed heterogeneity and low signal on DWI，but the recurrence group showed high signal.The mean ADC values in nasopharyngeal necrosis group and recurrence group were (1.073±0.0383)×10.3mm2/s and (0.844±0.0309)×10.3mm2/s respectively，the difference of which was statistically significant (t=4.645，P＜0.001). The maximal values in these two groups were (1.728±0.0527)×10.3mm2/s and (1.477±0.0675)×10.3mm2/s respectively，the difference of which was also statistically significant (t=2.932，P＜0.005). Conclusions DWI and ADC value are important in differential diagnosis of nasopharyngeal necrosis and tumor recurrence after radiotherapy.DWI and ADC can be used as important supplementary to routine MRI. Related Articles | Metrics

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Expressions of CK19 mRNA and CK20 mRNA in peripheral blood of patients with laryngeal squamous cell carcinoma at prior-and post-treatment stages MENG Qing-Xiang, XIE Jing-Hua, GAO Xiong-Hui, LI Ming, WANG Lei, LI Peng 2011, 3 (4):  302-305.  doi: 10.3969/j.issn.1674-5671.2011.04.09 Abstract ( 433 )   PDF (716KB) ( 480 )   Save Objective To study the expressions of CK19 mRNA and CK20 mRNA in peripheral blood of patients with laryngeal squamous cell carcinoma before and after radical treatment，and its clinical significances.Methods The nested reverse transcription polymerase chain reaction (nested RT-PCR) was used to detect the expressions of CK19 mRNA and CK20 mRNA at different stages in 48 cases of laryngeal squamous cell carcinoma. Dynamic observation of the expressions of CK19 mRNA and CK20 mRNA was applied within 24 months after operation. Results The positive expressions of CK19 mRNA and CK20 mRNA in peripheral blood at prior-treatment stage of 48 cases were 37.5% (18/48) and 45.8% (22/48) separately.The positive expressions of CK19 mRNA and CK20 mRNA were decreased statistically within 24 months after treatment (P＜0.05)，but these phenomena did not show in the cases with local recurrence.The positive expressions of CK19 mRNA and CK20 mRNA at prior-treatment in the patient with local recurrence were statistically higher than that without local recurrence.Conclusion The dynamic detections by nested RT-PCR for CK19 mRNA and CK20 mRNA in peripheral blood of patients with laryngeal squamous cell carcinoma at prior-and post-treatment stages could be used to evaluate micrometastasis and clinic treatment，and to predict the risk of local recurrence in the level of molecular biology. Related Articles | Metrics

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Expression of VEGF-C and COX-2 in bronchioalveolar carcinoma and its significance ZOU Liang, GAO Hui-Chun, ZHOU Jian-Hong, FENG Qiong 2011, 3 (4):  306-309.  doi: 10.3969/j.issn.1674-5671.2011.04.10 Abstract ( 460 )   PDF (1020KB) ( 501 )   Save Objective To investigate the expression and clinical significance of VEGF-C and COX-2 in bronchioalveolar carcinoma (BAC). Methods 60 cases of BAC were enrolled in experimental group；the control group contained 20 cases of pulmonary adenocarcinoma with BAC features and 22 cases of pulmonary adenocarcinoma. Expressions of VEGF-C and COX-2 were detected by immunohistochemical. Results Positive rates of VEGF-C in BAC，pulmonary adenocarcinoma with BAC features and pulmonary adenocarcinoma were 66.7%，90.0% and 95.5% respectively，the difference of which was statistically significant (P＜0.01). The positive rates of VEGF-C were notably higher in non-mucinous BAC than that in mucinous (P＜0.01)，and in lymphonode metastasis than in that without lymphonode metastases (P＜0.05). The positive rate of VEGF-C was not correlated with patients’ gender and age，and the localization，size and TNM stage of tumor (P＞0.05).  Positive rates of COX-2 in BAC，pulmonary adenocarcinoma with BAC features and pulmonary adenocarcinoma were 63.3%，75.0% and 77.3% respectively，the difference of which was not statistically significant (P＞0.05). The positive rates of COX-2 were obviously higher in lymphonode metastasis than that without lymphonode metastases (P＜0.05)，in tumor with diameter >3cm than that with diameter <3cm (P＜0.05). The positive rate of COX-2 was not correlated with patients’ gender and age，and the localization，size，histological type and TNM stage of tumor (P＞0.05).The expression of VEGF-C was positively correlated with that of COX-2 (r=0.269，P＜0.05). Conclusion Combined detection of VEGF-C and COX-2 could be applied in assessing and predicting the invasion and metastasis of BAC. Related Articles | Metrics

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The safety of implanting slow-releasing 5-fluorouracil in abdominal cavity during the operation on hepatocellular carcinoma TANG Yang-Yang, LIN Xin-Ju, PENG Min-Hao, PENG Tao, XIAO Kai-Yin 2011, 3 (4):  309-312.  doi: 10.3969/j.issn.1674-5671.2011.04.11 Abstract ( 466 )   PDF (716KB) ( 862 )   Save Objective To study the safety of regional implant of slow-releasing 5-fluorouracil microparticles into abdominal cavity during the operation on hepatocellular carcinoma (HCC).Methods  Patients with radical resection of HCC were divided randomly into two groups. Treatment group were implanted slow-releasing 5-fluorouracil microparticles into abdominal cavity, the control group were without intraperitoneal chemotherapy.The blutbild, functions of liver and kidney, blood coagulation, peritoneal drainage, length of hospital stay and postoperative complications were observed.Results There were no significant difference on the indexes of blutbild, blood coagulation, functions of liver and kidney between the two groups (P＞0.05).The local pain, incidence of pleural effusion and volume of abdominal drainage were significantly higher in the treatment group than that of control group (P＜0.05).The incidences of other postoperative complication and length of hospital stay were not significantly different (P＞0.05).Conclusions Using the slow-releasing 5-Fluorouracil microparticles in patients with HCC can improve the effect of regional chemotherapy.The treatment is with less impact on organ-functions, less post-operative complications and?good safety. Related Articles | Metrics

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Influence of anemia on treatment efficacy and survival prognosis of patient with advanced non-small lung cancer ZHAO Wen-Hua, SONG Xiang-Qun, YU Qi-Tao, ZENG Ai-Ping, HE Jian-Bo, WANG Hui-Lin, NING Rui-Ling, ZHOU Shao-Zhang 2011, 3 (4):  312-315.  doi: 10.3969/j.issn.1674-5671.2011.04.12 Abstract ( 548 )   PDF (710KB) ( 625 )   Save Objectives To investigate the correlation between hemoglobin level and chemotherapy efficacy as well as survival prognosis in patients with unresectable non-small-cell lung cancer (NSCLC). Methods The clinical data of 280 patients with unresectable NSCLC who were admitted in the Guangxi Tumor Hospital from January 2005 to January 2007 were retrospectively analyzed.The patients were divided into anemia group and non-anemia group，according to their gender，clinical stage and histopathological type. Statistical difference was analyzed by χ2 test. Survival analysis was evaluated by Kaplan-Meier and log-rank test.  Multivariate analysis was performed by Cox stepwise regression model. Results The clinical benefit rate of chemotherapy in the anemia group was decreased (P＜0.05)，and the survival time in non-anemia group was longer than that of anemia group (P＜0.001). Between the two groups，the survival time of stage ⅢB/Ⅳ，adenocarcinoma and   undifferentiated  carcinoma was statistically different (P＜0.05)，but that of stage ⅢA，squamous carcinoma and adenosquamous carcinoma was not significantly different (P＞0.05). The Cox analysis showed that anemia was an independent prognostic factor for NSCLC patient. Conclusions Hemoglobin level before initial therapy is related to survival time and the clinical benefit rate of patients with NSCLC. Anemia is an independent prognostic factor for NSCLC patients. Related Articles | Metrics

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Effect of capecitabine combined with vinorelbine on treating the patients with relapsed and metastatic breast cancer LU Yong-Kui, XIE Wei-Min, ZHOU Wen-Xian, HU Xiao-Hua, TAN Xiao-Hong, LIAO Xiao-Li, LIU Zhi-Hui, LI Yong-Qiang, CEN Hong 2011, 3 (4):  315-318.  doi: 10.3969/j.issn.1674-5671.2011.04.13 Abstract ( 522 )   PDF (717KB) ( 598 )   Save Objectives To evaluate the effect and adverse reactions of capecitabine combined with vinorelbine on treating the recurrent and metastatic breast cancer that failed to the treatment of taxanes and anthracycline. Methods Thirty-two patients with recurrent or metastatic breast cancer were treated by capecitabine and vinorelbine.Vinorelbine was given at the dose of 25mg/m2 on day 1st and day 8th，and xeloda was given at the dose of 2 000mg/m2/d，twice a day for two weeks followed by one week of rest as a cycle. All patients were treated with two or more cycles.Results Among the 32 patients，five achieved complete response，12 achieved partial response. The total response rate was 53.1%，and the clinical benefit response (CR+PR+SD) was 87.5%.The common adverse reactions were hand-foot syndrome，skin pigmentation，nausea，vomiting，anorexia and fatigue.Mild moderate anemia and leukopenia were observed in 36.4% of patients. Mild elevation of total bilirubin and alanine transaminase (ALT) occurred in a few patients. Conclusion Capecitabine combined with vinorelbine is an effective ragmen for treating patients with relapsed and metastatic breast cancer，especially for those who have failed to chemotherapy with anthracycline and taxanes. The regimen is tolerable while has mild adverse reactions. Related Articles | Metrics

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Transrectal ultrasound-guided biopsy for diagnosing prostate cancer LI Xiang, LI Hang, ZHAO Sheng-Fa, YANG Wei-Ping, DING Zhan-Ling, LI Hong-Xue 2011, 3 (4):  318-320.  doi: 10.3969/j.issn.1674-5671.2011.04.14 Abstract ( 544 )   PDF (1164KB) ( 639 )   Save Objective To evaluate the value of transrectal ultrasound (TRUS)-guided biopsy for differential diagnosing prostate cancer (PCa) from other prostatic diseases.Methods Transrectal ultrasound-guided biopsy was performed on 35 patients whose digital rectal examination showed positive results，and TRUS results showed suspected abnormalities.Results The diagnostic sensitivity of TRUS-guided biopsy was 84.2%，and its specificity was 81.5%. Conclusion  Biopsy guided by TRUS is convenient and safe，and has important significance for clinical diagnosis. Related Articles | Metrics