Abstract:Objective:To establish a MDA-MB-231 human breast cancer cell line stably express GFP-C-Daam1, and to investigate the influence of Daam1 on the mobility of breast cancer cells. Methods:The lentivrial vectors containing GFP or GFP-C-Daam1 were constructed and infected human breast cancer cells MDA-MB-231. MDA-MB-231 cells were screened by puromycin and stable expression of GFP or GFP-C-Daam1 was confirmed by Western blotting. Boyden chamber assay was used to test the intrinsic motility of the cells. Results:The cell lines stably expressing GFP or GFP-C-Daam1 were established successfully by the lentivrial infection. The GFP-C-Daam1 expressing cells showed greater intrinsic motility, suggesting the products of the interesting gene were active and functional. Conclusion:MDA-MB-231 breast cancer cell lines stably expressing GFP or GFP-C-Daam1 were successfully established using lentiviral methods. These cell lines can be used as model cells for further study of active Daam1 targeting.

Abstract:Objective:To investigate the effect of prostaglandin E2 (PGE2) on expression and enzyme activity of matrix metalloproteinase 2(MMP2) by EP1 receptor in cholangiocarcinoma cells HuCCT1 cells. Methods:HuCCT1 cells were treated with PGE2, EP1 receptor agonist, EP1 receptor antagonist, protein kinase C(PKC) inhibitor and Ca2+ chelating agent, RT-PCR and gelatin zymography were employed to detect the mRNA level and enzyme activity of MMP2 in HuCCT1 cells. Results:The mRNA levels of MMP2 increased 89.14%(P < 0.01), 163.89%(P < 0.01); and the enzyme activity of MMP2 increased 69.11%(P < 0.05), 117.65(P < 0.01) compared with the control group after treated with 5 μmol/L PGE2, 5 μmol/L EP1 receptor agonist 17-PT-PGE2 respectively. The mRNA level and the enzyme activity of MMP2 in HuCCT1 cells treated with 10 μmol/L EP1 receptor antagonist sc-51322 decreased 47.74%(P < 0.05),84.58%(P < 0.01) compared with the cells treated with PGE2. After treated with 5 μmol/L PGE2 or 5μmol/L 17-PT-PGE2, the enzyme activity of MMP2 in EP1R-pcDNA3 transfected HEK293 cells increased 36.07%(P < 0.05), 61.59%(P < 0.05) compared with the control group. When treated with 5 μmol/L PKC inhibitor BIS-1, 10 μmol/L Ca2+ chelating agent BAPTA-AM, the mRNA levels of MMP2 decreased 44.17%(P < 0.05),34.42%(P < 0.05); and the enzyme activities of MMP2 decreased 70.95%(P < 0.05),71.82%(P < 0.05) compared with the cells treated with 17-PT-PGE2. Conclusion:PGE2 might up-regulate the mRNA level and the enzyme activity of MMP2 through EP1 receptor in cholangiocarcinoma HuCCT1 cells, which could be partly related to the Ca2+/PKC signaling pathway.

Abstract:Objective:To isolate, purify, culture and identify the carcinoma-associated fibroblasts(CAFs) from the human epithelial ovarian neoplasm. Methods:The primary ovarian CAFs were obtained from human epithelial ovarian neoplasm and purified by trypsin digestion and adherence repeatedly. The cells were verified according to morphological observation and immunocytochemical staining of certain proteins. Additionally, their proliferation indexes were assayed. Results:The purified ovarian CAFs were obtained successfully. The typical morphology of ovarian CAFs was simple fusiform fiber like cells. The ovarian CAFs exhibited lower proliferation than tumor cells and showed positive staining for fibroblast specific protein-1, vimentin, desmin and especially for α-smooth muscle actin, and negative staining for cytokeratin(broad-spectrum), cytokeratin 7, cytokeratin 20, CD31 and CD90. Conclusion:The primary cultured CAFs derived from the human ovarian carcinoma tissue can be obtained successfully with digestion method. This provides us an experimental foundation for further studies on the roles of CAFs in the initiation and progression of the cancer.

Abstract:Objective:To investigate the inhibitory effect of monoclonal antibody(McAb) NJ488-1 on the lung adenocarcinoma cell line SPC-A1 in vivo and in vivo. Methods:SPC-A1 cells were plated on a soft agar matrix and treated with McAb NJ488-1 (0, 200, 400, 800, 1600, 2000 μg/ml). After 2 weeks’ incubation, the colony formation efficiency and the inhibition ratio of colonies were calculated. Xenograft was established by subcutaneous injection of SPC-A1 cells and antibodies were administered intraperitoneally at three different doses (200, 400, 800 μg per mouse). Animals were monitored by tumor size. Three weeks after the initiation of treatment, tumors were removed and weighed, and tumor growth inhibition was calculated. SPC-A1 cells were cultured with or without 400 μg/ml McAb for 24 h or 48 h. The morphology changes of cells were observed and the apoptosis rates were measured by flow cytometry. Results:In soft agar assay, the colony formation efficiency in McAb groups reduced in a dose-dependent manner. The higher level of antibody inhibited colony formation compared to the lower one, with 62.5% inhibition ratio at 400 μg/ml vs 23.4% at 200 μg/ml. When the concentration of McAb was 800 μg/ml or even higher, the inhibition ratio was up to 100%. McAb caused varying degrees of decrease in tumor volume compared with control mice(P = 0.004, P = 0.003, 400 μg and 800 μg McAb group vs control). In the 200, 400, 800 μg McAb groups, tumor growth inhibition was 10.44%, 37.29% and 44.04%, respectively. The average tumor weights of the 400, 800 μg McAb group and the control group, the 200 μg McAb group and 800 μg McAb group were significantly different(P = 0.032, P = 0.015, P = 0.048, respectively). NJ488-1 led to the obvious cyto-morphology changes and significantly induced the apoptosis of SPC-A1 cells in a time-dependent manner(P < 0.001). Conclusion:The McAb NJ488-1 inhibited the growth of lung adenocarcinoma cell line both in vitro and in vitvo. It is suggested that cell apoptosis induced by NJ488-1 might be the mechanism of the anti-tumor activity.

Abstract:Objective:To identify the expression of leptin receptor in breast cancer cell line MCF-7, and to explore the effects and mechanisms of leptin on the anti-proliferative effect of tamoxifen on MCF-7. Methods:The expression of leptin receptor was detected by immunofluorescence and Western blot. After treated with 100 ng/ml of leptin and 1 000 nmol/L of tamoxifen(TAM), cell proliferation and the levels of estrogen receptor α(ERα) in MCF-7 were examined. Plasmid pERE-LUC which containing estrogen response element(ERE) and reporter gene luciferase was constructed and transfected into MCF-7 cells, the expression of luciferase was detected to reflect the effects of leptin on the transcription of ERα. Results:Leptin receptor was positively expressed in MCF-7 cells. Leptin weaken the inhibition of TAM on MCF-7 proliferation by upreglulating the transcription of ERα. Conclusion:Leptin could interfere with the inhibition of TAM on breast cancer cells, it may contribute to the efficacy of tamoxifen treatment for breast cancer patients.

Abstract:Objective:To prepare 5-fluorouracil(5-FU) magnetic liposome with Fe/Fe3O4 core-shell structural magnetic nanoparticles(FCSN) and analysis its characteristics. Methods:Fe3O4 nanoparticles were used to prepare FCSN by reduction-oxidation method. The characteristics of FCSN were determined by X-ray diffraction (XRD), transmission electron microscopy (TEM) and vibrating specimen magnetometer (VSM). Using reverse-evaporation method to prepare 5-FU-FCSN magnetic targeted liposome and the best formula of magnetic liposome was obtained through the four factors of three levels orthogonal experiment. The characteristics of magnetic liposome were measured with particle size analyzer(PCS) and TEM. Entrapment efficiency was determined by gel-chromatography. Results:FCSN was similar to a globe with the average particle size of 70 nm. XRD and TEM observation of FCSN shown that its shell was Fe3O4 and kernel was Fe, magnetic saturation intensity was 107.54 emu/g. 5-FU-FCSN magnetic liposome was similar to a globe with the average particle size of 202.5 nm, the entrapment efficiency of 33.5%. 5-FU-FCSN magnetic liposome was relatively stable in 4℃ refrigerator after a month. Conclusion:5-FU-FCSN magnetic liposome was successfully prepared, which can be stored in 4℃ refrigerator for a long time.

Abstract:Objective: To explore the mechanism of large-scale transcriptional silence during germinal vesicle (GV) oocytes maturation in mice. Methods:The separated GV oocytes were obtained from 4~6-week-old ICR female mice. As for the observation about the expressional and distributional changes of both markers in the process of the chromatin configuration from non-surround nucleolus(NSN) type into surround nucleolus(SN) type in mice GV oocytes, these oocytes were immunofluorescence stained with RNA polymeraseⅡ antibody(anti-RNA PolⅡCTD, clone 4H8) and the second serine phosphorylated antibody at RNA polymeraseⅡCTD group (anti-RNA PolⅡCTD Ser2-P, clone H5); as well as histone H3K4/H3K9 tri-methylation antibodies (anti-H3K4me3, anti-H3K9me3). Results:In NSN type GV oocytes, RNA CTD PolⅡSer2-P was distributed with characteristic plaques shape in nucleus; meanwhile, H3K4me3 also presented the strong positive signals in nucleus. However, in SN type oocytes, RNA CTD PolⅡSer2-P lost the characteristic distribution and diffused in nucleus; moreover, the positive signal in nucleus showed by H3K4me3 disappeared. Conclusion:The reduction of RNA polymeraseⅡactivity and the change of histone modification status played an important role in the large-scale transcriptional silencing during germinal vesicle oocytes maturation of mice.

Abstract:Objective:To explore the regulatory effects of lactic acid bacteria on the secretion of tumor necrosis factor(TNF)-α, interleukin(IL)-12 p40 and transforming growth factor(TGF)-β in THP-1 cells induced by lipopolysaccharide(LPS). Methods: THP-1 cells were firstly stimulated by phorbol 12-myristate 13-acetate(PMA) to differentiate into macrophage-like cells, and were secondly treated by LPS to induce inflammatory cytokines. Six groups were designed, including blank control group, LPS-treated group, L.para-treated alone group, L.para plus LPS-treated group, L.acid-treated alone group and L.acid plus LPS-treated group. The levels of TNF-α mRNA in THP-1 cells were determined by RT-PCR. TNF-α, IL-12 p40 and TGF-β in the supernatant of THP-1 cells were assayed by ELISA. The subcellular localization of NF-κB p65 was observed by immunofluorescence, while the quantitation of NF-κB(p65/p50) in THP-1 nucleus was detected by TransAMTM NF-κB kit. Results:① The secretions of TNF-α, IL-12 p40 and TGF-β in PMA-induced macrophage-like THP-1 cells were remarkably increased when treated by LPS, the same as L.para or L.acid treated alone respectively. However, TNF-α and IL-12 p40 in L.para-treated alone group were lower than those in LPS-treated group, whereas TNF-α and IL-12 p40 in L.acid-treated alone group were similar to the LPS-treated group. TGF-β induced by L.para or L.acid was strikingly higher than that of LPS. Interestingly, TNF-α and IL-12 p40 in PMA-induced macrophage-like THP-1 cells stimulated by LPS were notably inhibited when cells were treated with L.para plus LPS or L.acid plus LPS. TNF-α and IL-12 p40 in L.para plus LPS-treated group were much lower than in LPS-treated group. No inhibition of TGF-β were observed in PMA-induced macrophage-like THP-1 cells when treated with both lactobacilli.② Similar to result one, phosphorylation of IκB-α and nuclear NF-κB(p65/p50) in PMA-induced macrophage-like THP-1 cells increased in LPS-treated group, L.para-treated alone group and L.acid-treated alone group, while non-phosphorylation of IκB-α accordingly decreased. The regulatory effect of L.para was a little weaker than L.acid. Phosphorylation of IκB-α and nuclear translocation of NF-κB were significantly reduced in PMA-induced macrophage-like THP-1 cells when treated with L.para plus LPS or L.acid plus LPS. Conclusion:L.para and L.acid can inhibit the secretion of LPS-induced inflammatory cytokines TNF-α and IL-12 p40 when they interacted with LPS respectively. The negative regulation of lactic acid bacteria was related to inhibition of NF-κB signaling. Besides, the regulatory effects of lactic acid bacteria were evidently different among species.

Abstract:Objective:To study neural cell tropism of human herpesvirus 6A(HHV-6A) and its effect on neural cell cycle. Methods:The cell morphology was assessed by a light microscope. The fragment of HHV-6A U22 gene was amplified by PCR and relative amount of HHV-6A U22 gene was analyzed using quantitative real-time PCR. Expression of HHV-6A gB was examined by immunocytochemical and Western blot analysis. Cell proliferation was measured by MTT assay. Cell cycle analysis was evaluated by the propidium iodide assay. Results:On the fifth day after being infected by HHV-6A,U373 cells had no obvious change,but SK-N-SH and SHG44 cells showed cytopathic effect. HHV-6A U22 gene was detected in all three infected cell lines and its relative amounts kept reducing all the time. HHV-6A gB was positive in the three infected cell lines,furthermore, the expression levels in U373 and SHG44 were more than those in SK-N-SH. HHV-6A stimulated the proliferations of U373 and SHG44 cells,however,inhibited the proliferation of SK-N-SH cells significantly. Compared with uninfected cells,percentages of U373 and SGH44 cells were decreased in phase G1 but increased in phase S and G2 after cells were infected by HHV-6A,as to SK-N-SH,the percentages were increased in phase G1 but decreased in S and G2 phases. Conclusion:All three neural cell lines can sustain replication of HHV-6A,and moreover,HHV-6A can boost astrocyte proliferation and inhibit neuronal cell proliferation.

Abstract:Objective:To investigate the effect of 17β-estradiol (E2) on cultured cardiomyocyte hypertrophy induced by mechanical stretch. Methods:Cardiomyocytes from neonatal rats were cultured in vitro and cardiomyocyte hypertrophy was induced by mechanical stretch. The effect of E2 on cardiomyocyte hypertrophy was evaluated by the surface areas of cardiomyocytes using immunocytochmistry and the β-MHC protein levels detected by Western blot. The integrin β1 protein levels were determined by Western blot. Results:Mechanical stretch for 24 h significantly increased cell surface areas and β-MHC protein levels, which suggested that mechanical stretch is a direct stimulus for isolated cardiomyocytes to develop hypertrophy. Mechanical stretch also significantly increased the expression of integrin β1 protein in cultured cardiomyocytes. Pretreatment with 100 nmol/L E2 for 30 min significantly attenuated the increases in cell surface areas and β-MHC protein levels induced by mechanical stretch which were attenuated by estrogen receptor antagonist ICI182780. Furthermore, E2 also decreased the levels of integrin β1 protein induced by mechanical stretch, and this was also attenuated by ICI182780. Conclusion:E2 prevents cardiomyocyte hypertrophy and the increase of integrin β1 protein induced by mechanical stretch in vitro.

Abstract:Objective:To explore the effect and mechanism of grape seed proanthocyanidin extract(GSPE) on airway inflammation and hyperresponsiveness in a murine model of allergic asthma. Methods:BALB/c mice(n=32; aged 8 weeks) were randomly and averagely divided into 4 groups: control group, asthma group, budesonide group and GSPE group. The mice were sensitized and challenged with ovalbumin(OVA) to produce a murine model of asthma. The GSEP group mice were intraperitoneally injected with GSPE[50 mg/(kg·d)]. Mice in the budesonide group were treated with aerosolized budesonide. The airway responsiveness of mice in each group to acetylcholine chloride (Ach) was evaluated. HE staining and PAS staining were used to observe airway inflammation in lung tissue and airway mucus secretion, respectively. Quantification of cytokines in bronchoalveolar lavage fluid(BALF) and total serum immunoglobulin E(IgE) were detected by ELISA. The protein expression of inducible nitric oxide synthase(iNOS) in lung tissue was evaluated by Western blot analysis. Results:As compared with the control group, the asthma group showed significant increase of airway resistance, total inflammatory cell and eosinophil counts and interleukin(IL)-4 level in BALF, total IgE level in serum, as well as iNOS expression, while the level of interferon (IFN)-γ in BALF decreased (P < 0.05). Remarkable reduction was observed in all the above items except IFN-γ level, which increased, in the GSEP group and the budesonide group(P < 0.05). Conclusion:GSPE decreases the progression of airway inflammation and hyperresponsiveness by downregulating the iNOS expression, promising to be a potential drug in the treatment of allergic asthma.

Abstract:Objective:To clone the rabies virus G protein(RVG) gene fragments into prokaryotic expressing vector, and to express and purify RVG, which could be applied as diagnostic antigen for indirect ELlSA assay to detect rabies virus antibodies. Methods:The G gene of rabies virus was amplified by RT-PCR with a pair of specific primers designed according to the relevant sequence of GenBank. The PCR product was cloned into prokaryotic expression vector pGEX-6P-1 to construct the expression plasmid pGEX-RVG. The positive recombinant plasmid was transformed into host bacteria E.coli BL21(DE3) and induced with IPTG. In order to determine the optimal induction condition for soluble expression, optimization condition was studied. The glutathione S-transferase (GST) affinity chromatography was used to purify the recombinant protein, and the biological activity of RVG was analyzed by Western blot. Results:The prokaryotic expression vector pGEX-RVG were successfully constructed. After inducted by IPTG, the fusion protein about 36 000 was observed,which was the same as expected. The recombinant fusion protein has a fine biological activity which was confirmed by Western blot. Conclusion:The rabies virus G protein was successfully expressed and purified, which could be provided diagnostic antigen for indirect ELlSA assay for the detection of rabies virus antibodies.

Abstract:Objective:To investigate whether rat adipose derived stem cells could be induced into Schwann cell-like cells in vitro. Methods:Rat adipose-derived stem cells (ADSCs) were isolated, cultured, and multi-potential differentiation capacity of rat ADSCs were analyzed. The neurospheres were induced from rat ADSCs with epidermal growth factor (EGF) and basic fibroblast growth factor(bFGF). Surface markers were detected in neurospheres. Retinoic acid, forskolin, platelet-derived growth factor(PDGF) and heregulin were used to induce neurospheres into Schwann cell-like cells. The morphology was observed and surface markers were detected in Schwann cell-like cells. Results:Rat ADSCs were mono-layer fibroblastic cells, and were nestin negative and fibronectin positive and could undergo osteogenic and adipogenic differentiation. Rat ADSCs could be induced into floating neurospheres which were nestin positive and fibronectin negative. Neurospheres could differentiate into Schwann cell-like cells which were spindle like in shape and could express classic Schwann cell markers glial fibrillary acidic protein(GFAP), S100 and p75. Conclusion:Rat ADSCs could differentiate into Schwann cell-like cells in vitro. Schwann cell-like cells may be useful for the treatment of neurological diseases.

Abstract:Objective:To investigate the effect of mancozeb on T-type calcium currents(ICaT) in spermatogenic cells. Methods: Ca2+ currents were obtained in acutely dissociated mouse spermatogenic cells by the whole-cell patch clamp technique and the effects of mancozeb on ICaT were observed. Results:Mancozeb at the concentrations of 10, 20, 40, 80 and 120 μmol/L significantly inhibited ICaT in the mouse spermatogenic cells, with the K50 value of 35.60 μmol/L and the inhibition rates of mancozeb were(9.93±0.90)%,(20.93±2.50)%,(25.92±2.80)%,(42.64±3.10)% and (56.80±3.20)%, respectively(n=6, P < 0.05). Mancozeb of 120 μmol/L significantly changed the activation and inactivation of ICaT: the half activation potential(V1/2a) and the activation steepness factor (κa) from (-47.09±1.05)mV and (8.67±0.78)mV to (-51.46±0.84)mV and (10.56±0.619)mV, respectively(n=5, P < 0.05); the half inactivation potential (V1/2i) and the inactivation steepness factor (κi) from (-62.96±3.36)mV and (9.93±1.41)mV to(-64.11±5.55)mV and (13.64±1.95)mV, respectively (n=5, P < 0.05). Conclusion:Mancozeb has significant inhibitory effects on ICaT in mouse spermatogenic cells with concentration dependence.

Abstract:Objective:To investigate the mechanism of vascular endothelial growth factor(VEGF) expression in osteoblasts induced by durative pressure. Methods:Osteoblasts were isolated from the calvarias of neonatal(12 days old) Sprague-Dawley rats, cultured and identified according to published protocols. The cells were pretreated with or without PD98059 and cultured for 0.5, 2.0, 6.0 h under 100 kpa of pressure in sealed containers. The VEGF concentrations secreted into DMEM were detected by ELLSA. The mRNA levels of VEGF were evaluated by RT-PCR. The total ERK1/2 and the phosphorylation of ERK1/2 in osteoblasts were detected by Western blot. Results:The durative pressure increased the expression and secretion of VEGF, and meanwhile the phosphorylation of ERK1/2 in osteoblasts increased significantly. Pretreated the osteoblasts with PD98059 inhibited the phosphoryation of ERK1/2 and reduced the the expression and secretion of VEGF significantly. Conclusion:Durative pressure induces the expression of VEGF in osteoblasts by the activation of ERK1/2.

Abstract:Objective:To evaluate the prevalence of anti-tuberculosis drug resistance (DR) among tuberculosis (TB) patients in Jiangsu province and provide statistical data for government to prevent and control TB. Methods:The TB patients in Jiangsu province were sampled and drug susceptibility test(DST) was carried out according to the “Guidelines for surveillance of drug resistance in tuberculosis” developed by WHO/IUATLD. Results:①The DST results showed that 40.96% Mycobacteria tuberculosis strains were resistant to at least one drug we tested, and the proportion was 32.80% among new cases and 55.90% among previously treated ones. The prevalence of multidrug-resistant(MDR) TB was 16.61%, with a significant difference in new cases (7.63%) and in previously treated cases(33.07%). ② Patients in middle-age or with lower education background or living in the middle or north part of Jiangsu province had higher risks for DR or MDR. Conclusion:The high prevalence of drug resistance in Jiangsu province has been a major challenge for TB control.

Abstract:Objective:To evaluate the current situation of professional exposure to human immunodeficiency virus(HIV) in order to discuss the measure of postexposure prophylaxis Methods:The cases of occupational exposure to HIV in 2003~2009 were investigated and the data were statistically analyzed. Results:Among the totally 72 cases, there were 41 cases with acupuncture or incised wound, 10 cases with grab or bite wound, 15 cases were exposed by skin while 6 cases were exposed by mucosa. Fifty six cases were taken place in medical health institutions, 2 cases were happened in centers of disease prevention and control, 9 cases were occurred in public security agencies, one case was happened in blood transfusion services and the other 4 cases were belong to other occasions. Sixty one cases were local processed immediately after exposure, 42 cases took medicines with basic procedure, and 14 cases were treated with reinforcement procedure while the other 16 cases hadn’t taken any measures because of individual negative judgment. All cases were HIV antibody negative in the follow test. Conclusion:The risk of HIV infection after exposure to HIV in occupational occasions was relatively low. However, the consciousness of prevention should be strengthened to avoid the occurrence of blood transfusion diseases.

Abstract:Objective:To study the changes of serum levels of interleukin-6(IL-6) and C-reactive protein(CRP) in patients with gastric carcinoma and the relationship of the expression of vascular endothelial growth factor(VEGF) and CD31 with the serum level of IL-6. Methods:IL-6 was evaluated by enzyme-linked immunosorbent test(ELISA). CRP was detected by Beckman albumen measuring system. The expression of VEGF and CD31 in gastric carcinoma tissues was detected by immunohistochemistry. Results:The serum level of IL-6 in patients with gastric carcinoma was significantly higher than controls(P < 0.001) and concerned with staging. After operation, the serum level of IL-6 decreased significantly(P < 0.001). The percentage of VEGF positive cells and the microvessel density(MVD) labeled by CD31 antibody gradually increased with the staging. They were all positively correlated with the serum level of IL-6. Conclusion:The detection of serum IL-6-VEGF and CD31 in gastric carcinoma patients is useful to early diagnosis, therapy and to anticipate the condition and prognosis.

Abstract:Objective:To investigate the relationship of X-ray repair cross complement gene 1(XRCC1), poly (ADP-ribose)polymerase-1(PARP1) and apurinic/apyrimidinic endonuclease 1(APE1) polymorphisms with the efficacy of platinum-based chemotherapy for treatment of the patients with advanced non-small-cell lung cancer(NSCLC). Methods: A total of 151 patients with advanced NSCLC were routinely treated with platinum-based chemotherapy, and their clinical responses were evaluated. XRCC1 G28152A (Arg399Gln), XRCC1 C26304T (Arg194Trp), PARP1 T2444C (Val762Ala) and APE1 T1349G (Asp148Glu) of the patients were genotyped using the TaqMan method. The association of these polymorphisms with the clinical responses was analyzed using unconditional logistic regression model. Results:XRCC1 G28152A polymorphism was significantly correlated with clinical benefit. The efficacy of chemotherapy of XRCC1 28152 GA genotype carriers significantly increased compared with patients with GG genotype(adjusted OR=2.85, 95%CI:1.291~6.277, P < 0.05). Patients carrying XRCC1 28152 A allele(GA/AA) had better clinical response than patients with wildtype allele (adjusted OR=2.48,95%CI:1.330~6.075, P < 0.05). Compared with CC and CT genotypes, XRCC1 26304 TT genotype carriers had a roughly 64% decreased efficiency but with no statistical significance (adjusted OR=0.36,95%CI:0.040~3.298). A similar result was found in polymorphism of PARP1 T2444C,the response rate of chemotherapy of the patients with CC genotype significantly decreased compared with other (adjusted OR = 0.37, 95%CI:0.118~1.170). No significant association was found between APE1 T1349G polymorphism with clinical response. Conclusion: The XRCC1 G28152A (Arg399Gln) polymorphism is significantly associated with the clinical benefit of NSCLC patients receiving platinum-based chemotherapy, and the XRCC1 G28152A genotypes detected by TaqMan method may be useful in clinical applications to predict the sensitivity of chemotherapy in NSCLC patients.

Abstract:Objective:To compare perioperative effect of lobectomy by totally video-assisted thoracoscopic surgery(tVATS) or traditional open surgery(TOS) on immunologic function and prognostic factors of non-small cell lung cancer(NSCLC) patients. Methods:Forty NSCLC patients underwent lobectomy by TOS or tVATS were selected,20 patients in each group. The serum levels of interleukin-6(IL-6),insulin-like growth factor binding protein-3(IGFBP-3) and vascular endothelial growth factor (VEGF) of patients before surgery and at the 1st,3rd and 5th day after surgery were determined using the ELISA. At the same time,the clinical data were collected and compared between two groups. Results:There was no significant difference in the preoperative serum levels of IL-6,IGFBP-3 and VEGF between two groups. The levels of IL-6 and VEGF at the three postoperative time points in tVATS group were significantly lower than those in TOS group. The levels of IGFBP-3 at the three postoperative time points in tVATS group were significantly higher than those in TOS group. There were no death and no serious complications in two groups. No conversion to thoracotomy occurred in tVATS group. There was no significant difference in operative time,numbers of stations and numbers of lymph node dissected,the chest drainage days and drainage volume after surgery between two groups. In tVATS group,intraoperative blood loss,postoperative pain score,days of getting out of bed and postoperative hospital stay were significantly lower than those in TOS group. Conclusion:Comparing with TOS,the effect of lobectomy with tVATS on the immune suppression and the influence on tumor metastasis and recurrence is much less. It can improve the prognosis of patients,provide favorable conditions for the subsequent treatment.

Abstract:Objective:To explore the potential role of the surgical treatment in limited small cell lung cancer(SCLC) and analyze the factors affecting the prognosis. Methods:From January 1985 to December 2005, the survival rate of 70 in-patients with limited SCLC confirmed by histological pathology and imaging were analyzed. Furthermore, the related prognostic factors of patients underwent surgery were investigated by Kaplan-Meier estimator and Cox regression analysis. Results:The five-year survival rates of patients with stage Ⅰ,Ⅱ,Ⅲ SCLC were 63.8%, 40.1% and 33.2%, respectively. The Cox regression analysis indicated that TNM stage, application of platinum-based chemotherapy, operative manner and KPS scores were independent prognostic factors for SCLC patients after surgery. Conclusion: Patients with limited SCLC may benefit from the comprehensive treatment based on surgery. TNM stage, application of platinum-based chemotherapy, operative manner and KPS scores were independent prognostic factors for SCLC patients after surgery.

Abstract:Objective:To observe the effects of dezocine on the agitations after remifentanil-sevoflurane anaesthesia. Methods: Eighty ASAⅠorⅡ grade patients who were scheduled for total hysterectomy under laparoscopy,aged from 46 to 65 years,were randomly divided into experimental group(group E, n=40) and control group(group C, n=40). Patients in group E were administered with 10 mg dezocine and in group C with 2 ml normal saline at 20 min before the end of the operations. Heart rate(HR) and the mean arterial pressure(MAP) at the end of the operations, endotracheal extubation and 5 min after endotracheal extubation were recorded. Agitation score during endotracheal extubation and the Ramsay sedation score 5 min later were observed. Duration from the end of the operation to the extubation was also recorded. Results:HR and MAP at the end of the operations had no significant differences between two groups (P > 0.05),but they raised markedly in group C at extubation and 5 min later(P < 0.05). They were also obviously increased in group E at extubation(P < 0.05),but had no significant differences 5 min later(P < 0.05). There were remarkably differences in HR and MAP at extubation and 5 min later between group E and group C(P < 0.05).Agitation score was lower in group E than in group C during extubation(P < 0.05). Ramsay sedation score was higher in group E than in group C 5 min after extubation(P < 0.05). Duration from the end of operation to extubation had no statistic differences between the two groups(P < 0.05). Conclusion:Dezocine can effectively alleviate the agitations after remifentanil-sevoflurane anaesthesia.

Abstract:Objective:To investigate the association between methylenetetrahydrofolate reductase (MTHFR) 677C/T polymorphism and Alzheimer’s disease(AD). Method:A meta-analysis including 1 048 cases and 1 220 controls in eligible studies were performed. The odds ratio(OR) of genotype and allele frequencies were regarded as statistics value, a random-effect or fixed-effect model was used in the analysis, and publication bias was assessed. Results:Nine papers met the criterion and were included in the meta study, and no significant publication bias was showed. The OR(95%CI) values of T/C and(TT+TC)/CC were 1.21(1.07~1.37) and 1.44(1.07~1.93). Conclusion:The MTHFR C677T polymorphism is associated with AD.

Abstract:Objective:To analyze the clinical features of Gitelman syndrome(GS) in order to improve our understanding of this syndrome. Methods:Retrospective study was conducted on 14 cases of GS in past 5 years in Department of Endocrinology, the First Affiliated Hospital of Nanjing Medical University. Results:The age ranged from 13~54(32.9±12.0) years old with 5 males and 9 females. The history ranged from one month to 15 years. Average body mass index (BMI) was (21.0 ± 2.9) kg/m2 while average systolic blood pressure was (109.6 ± 10.8) mmHg and diastolic blood pressure was (69.6 ± 6.7) mmHg. Clinical symptoms included lower extremities weakness, recurrent paralysis, polydipsia, polyuria, nocturia and tetany. Laboratory findings included hypokalemia, hypomagnesaemia, excessive urine potassium, hypocalciuria, metabolic alkalosis. Recumbent-upright tests showed: supine and upright plasma renin activity (PRA) augmentation, angiotensin-Ⅱ (ATⅡ) elevation and hyperaldosteronism. Renal biopsies revealed juxtaglomerular apparatus hyperplasia (3/4). All symptoms were relieved after potassium and magnesium supplementation or with combined spironolactone. However serum potassium(11/14) and magnesium(10/14) levels were still below normal range. Conclusion:The clinical features of GS includes hypokalemia, hypomagnesaemia, hypocalciuria, hypotension and hyperreninemia, high level of ATⅡ, hyperaldosteronism and high pH in arterial blood gas analysis which can be shorten as “four hypos and four hypers”. The treatment of this syndrome includes potassium and magnesium supplementation, or combination with anti-aldosterone medications. Generally, these patients have good prognosis.

Abstract:Objective:To evaluate the changes of serum prostate specific antigen(PSA) levels in type 2 diabetes and hypertension patients. Methods:Totally 612 patients from 40 to 78 years old were divided into three groups: diabetes group, hypertension group and control group. All the patients were invited to complete a questionnaire and serum PSA test. Then a two-way ANOVA was performed to examine the association of type 2 diabetes, hypertension and PSA. Results:The serum PSA level of diabetes group was lower than that of control group and there was statistically significant difference between them(P < 0.05). The serum PSA level of hypertension group was higher than that of control group. But the difference between the two groups wasn’t statistically significant(P > 0.05). Conclusion:Our results suggest that the patients with type 2 diabetes experience lower serum PSA levels comparing with those without diabetes. Further research is needed to elucidate whether the difference results in a relatively lower less cancer detection among diabetic men.

Abstract:Objective:To measure the rabbit liver volume(LV) by volume-measuring method supplied by multi-slice CT (MSCT) software and evaluate it’s application value in liver transplantation. Methods:Ten New Zealand rabbits underwent plain and enhanced 3-phase scan using 16-slice CT. Rabbit LV was measured by both method for measuring outline and volume-measuring method supplied by MSCT software in vivo respectively, and followed by measurement of water displacement in vivo. Results:Rabbit LV was(61.00 ± 8.95) cm3 when measured by method for measuring outline, which was significant lower than (73.25 ± 7.49) cm3 when measued by measuring water displacement, whereas no significant difference was found between(73.30 ± 10.88) cm3 and(73.25 ± 7.49) cm3 when comparing rabbit LV measured by volume-measuring method supplied by MSCT software and measurement of water displacement. Conclusion:Measurement of water displacement is better for measuring rabbit LV than measuring outline.

Abstract:Objective:Comparison of different scanning and injection protocols for bronchial artery CT angiography(CTA). Methods:Eighty cases scheduled for bronchial artery CTA were randomly assigned into four groups according to the injection protocol. Cases in group 1 to group 3 adopted the injection rate of 4, 5 and 6 ml/s respectively, and used automatic threshold preset at 300 HU to trigger the initiation of the scan, with total contrast dose of 1.5 ml/kg body weight. Cases in the group 4 adopted an injection rate of 6 ml/s, triggering threshold of 120 HU, and 1.0 ml/kg body weight of contrast. High density contrast(350 mgI/ml)was used in all cases. The CT values of pulmonary artery, superior vena cava, aortic artery and bronchial artery before and after enhanced scanning were measured in the 4 groups. Numbers of cases with excellent image quality of bronchial artery display were also evaluated. Results:The CT values of pulmonary artery and superior vena cava among four groups hadn’t significant difference(P > 0.05). The CT values of aortic artery in group 4 were higher than other three groups significantly(P < 0.05). The CT values of bronchial artery in group 4 were higher than group 1 and group 2 significantly (P < 0.05). The excellent rate of bronchial artery CTA for group one to group four were: 50%, 55%, 65% and 80%, respectively. Conclusion:This research showed that the injection rate of 6 ml/s, triggering threshold of 120 HU, 1.0 ml/kg body weight of high density contrast dose provide higher contrast,and it’s on optimal protocol for bronchial artery CTA.

Abstract:Objective:To explore the CT findings and features of pancreatic serous cystadenoma, mucinous cystadenoma and mucinous cystadenocarcinoma. Methods:Clinical data and spiral CT findings of 26 cases with pathologically proven pancreatic cystic tumors were analyzed retrospectively. Results:Thirteen lesions were serous cystadenoma which showed round or lobulated cystic or cystic-solid tumor, fibrotic septum were seen in 7 cases and central dot calcification were found in 5 cases. Cystic wall and septum was enhanced in portal venous phase imaging. Eight lesions were mucinous cystadenoma, they showed round or irregular low density mass, and 2 of them were single cyst and 6 cases were multiple cyst. The cystic wall was uneven and calcification of cystic wall and septum were found in 2 lesions. Five lesions were mucinous cystadenocarcinoma. Wall nodules were relatively large. One case located in the pancreatic head had occurred pancreatic duct dilatation and with serum CA-199 increased. Internal calcification was detected in 1 case and peripancreatic fat space was obscured in 2 cases. Conclusion:The CT imaging can display the size, shape, internal characteristic of pancreatic cystadenoma and relationship with adjacent tissue, it provides important information for clinical diagnosis and therapy.

Abstract:Objective:To investigate clinical diagnostic value of transvaginal color Doppler ultrasound on atypical ectopic pregnancy. Methods:Transvaginal color Doppler ultrasound two-dimensional images, blood flow distribution and frequency spectrum characteristics of 32 atypical ectopic pregnancy patients confirmed by operative pathological diagnosis in past 4 years were retrospectively analyzed. Results:Transvaginal color Doppler ultrasound displayed atypical ectopic pregnancy lesions clearly: 20 cases (62.5%) with annex area pouch and solid bag piece, 9 cases (28.1%) with annex area solid bag piece, 3 cases (9.4%) had only oviduct local increase coarse swollen; 4 cases (1.3%) with intrauterine “false pregnancy sac” signs; 25 cases(78.1%) with and 7 cases(21.9%) without endometrial thickening; all cases with abdominal pelvic cavity accumulated fluid pine; nourish blood flow signal was detected in 29 cases(90.6%), pulsed Doppler spectrum showed the high-speed low resistance widened bloods tream spectrum, the resistance index was 0.40±0.07, 3 cases(9.4%) were failed to explore blood flow signal. Color Doppler ultrasound diagnostic accuracy rate was 81.3%(26/32). Conclusion:Transvaginal color Doppler ultrasound display images clearly and fast, don’t need to fill bladder. Early diagnosis of the atypical ectopic pregnancy based on ultrasonographic and blood flow spectrum performance characteristics has high accuracy rate, and should be a preferred choice in clinical examination.