Objective To study the inducing apoptosis of Hedyotis diffusa extract (HDE) on human hepatocarcinoma cell line Bel7402 and its molecular mechanism. Methods Human hepatocarcinoma cell line Bel7402 was used in vitro cell culture, there were 3 groups:control group, HDE group and 5-Fu group, the Bel7402 cells were dealed with them respectively. The apoptosis morphologic changes of human hepatocarcinoma cells were observed by invert microscope. The apoptosis rate was detected with hematoxylin stain by light microscope. Inhibition of Bel7402 cell proliferation was measured by MTT assay. The expression of oncogene Bcl-xl and anti-oncogene p53 of Bel7402 ceils were observed with RT-PCR assay. Results Compared with the control group, the condensation of nuclear, vacuolar degeneration of mitochondria could be found through light microscope. The apoptosis rate of HDE group was remarkably increased compared with that in control group (P

Objective To study the diagnos tic value of mammography, color Dopp ler ultrasound and biopsy in early breast cancer and to evaluate its diagnostic accuracy. Methods 55 patients with breast cancer an d 25 patients with benign breast masses, proved pathologically, were checked by mammogra phy, color Doppler ultrasound and biopsy. Results Mammography demonstrated bre ast cancer in 45 cases, the sensitivity and specificity were 82.0% and 88.0%, re spectively, with the accuracy of 83.8%; Color Doppler ultrasound revealed breas t cancer in 43 cases, the sensitivity, specificity and accuracy were 78.2%, 84 .0% and 80.0%; Biopsy demonstrated breast cancer in 53 cases, the sensitivity, specificity and accuracy were 96.4%, 92.0% and 95.0%. When ultrasound, mammagrap hy and biopsy were combined together, the sensitivity, specificity and accuracy were 98.2%, 96.0% and 97.5%. Conclusion Mammography, color Dopp ler ultrasound combined with biopsy can increase the diagnostic sensitivity and accuracy of ear ly breast cancers.

Objective:To evaluate the effect of gtfB specific antisense phosphorothioate-modified oligodeoxyribonucleotides(PS-ODNs) on S.mutans sucrose-dependent adherence.Methods:Antisense oligodeoxyribonucleotide targeted to gtfB sequence 709~726 bp(PS-ODN1) and 3 479~3 497 bp(PS-ODN2) were synthesized.Natural genetic transformation of S.mutans with PS-ODN1 and PS-ODN2 was respectively performed.Adhesion of S.mutans to saliva coated hydroxyapatite was examined by crystal violet staining and destain with absolute ethanol.The absorbance at 620 nm was measured by plate reader(the absorbance value derived from the wells without sucrose was used as background and was subtracted).Results:The adhesion ability of the strains treated with antisense PS-ODN was significantly lower than that of the control(P

0.05).Conclusion:So-fastTM can greatly improve the penetration of PS-ODN into S.mutans and can be used as appropriate delivery system of PS-ODN for S.mutans. No matter what approaches were adopted, the uptake rate reached the maximum at 5 ?mol/L and 2 h-exposure. The penetration can not be enhanced by increasing the PS-ODN concentration and the transformation time.

Objective To investigate the effect of expression level and methylation level of miR-200b on proliferation, invasion and apoptosis of gastric cancer cell line MGC-803 in vitro. Methods Normal human gastric epithelium cell line GES-1, and gastric cancer cell line MGC-803 cells were cultured and harvested to extract total RNA. Then miR-200b ex?pression level was examined via q-PCR;Methylation in promoter of miR-200b was revealed by Bisulphite PCR. MGC-803 cells were treated with different concentrations of 5′-Aza-CdR to test its effect on miR-200b expression and methylation of its promotor. The effect of its treatment at 10μmol/L for 72 h on invasion,proliferation and apoptosis of both cell lines were detected by Transwell assay, Flow cytometry and apoptosis assay respectively. The gene related to EMT (epithelial-mesen?chymal transition ) such as E-cadherin,N-cadherin,ZEB1,Slug were checked by Western blot. Results The expression of miR-200b in GES-1 is higher than that in MGC-803(P=0.022). The methylation level in promoter region of miR-200b in GES-1 is lower than that in MGC-803(P=0.034). After treated with 5′-Aza-CdR, the expression of miR-200b in MGC-803 cell was up-regulated in a timely dependent manner. On the contrary, the methylation in promoter of mirR-200b were down-regulated when 5′-Aza-CdR were added at 10μmol/L(P=0.043). What’s more, 5′-Aza-CdR administration increas?es cell apoptosis especially in aged cells and delays cell cycle at G0/G1 which in turn decrease ratio of cells in S phages. 5′-Aza-CdR treatment also decreased invasiveness and induced expression of E-cadherin, as well as down regulated the ex?pressions of N-cadhrin, ZEB1, Slug and MMP9 in MGC-803 cells. Conclusion Expression of miR-200b could be affected by methylation of promoter of miR-200b and in turn regulate proliferation and invasion of gastric cells in vitro.

AIM To establish anti-osteosarcoma antibody producing hybridoma cell lines and to study the characterization of the monoclonal antibodies. Methods BALB/c mice were immunized with human osteosarcoma cells OS-9607 and the immunized spleen cells were fused with SP2/0 cells to raise hybridoma. The propert of antibody and it's cytotoxic effect were studied respectively with immunohistochemistry methods using OS-9607 and normal hepatocytes、 Western Blot methods and MTT method. Results A hybridoma cell line named 3D9 was established and it secreted high quality mAbs steadily. 3D9 cell had all the characteristics of hybridoma. The mAb's corresponding antigens was specifically and highly expressed in human osteosarcoma. With enzyme-labeled immunohistochemical staining on formaldehyde -fixed sections from human osteosarcoma,it was found that 83％ of the specimens expressed the corresponding antigen. Most of them were expressed on the nuclear of cells, no positive expression was observed in kinds of normal tissues. Western Blot showed 3D9's corresponding molecule weight is Mr54 000. MTT assay proved that the cytotoxicitis of effective groups were higher than control groups. Conclusion A high quality hybridoma is cultured and the mAb secreted by it has osteosarcoma specificity and obvious cytotoxic effect. It may be a new biochemical mark of osteosarcoma, and it's clinical prospect of immunotherapy will be wide.

Objective To study the clinical significance and change law of the serum inflammatory factor in the elder patients with femoral neck fracture during perioperative period. Methods 62 patients with femoral neck fracture were selected as research object,and the serum IL-1,IL-6,IL-10,TNF-α,CRP and BGP of the patients at preoperative day and after the treatment at the first,third,seventh and fourteenth day were detected with ELISA. Results The serum IL-1 ,IL-6,IL-10,TNF-α and CRP went up first and then down,the levels were highest at the third day, and they were compared to the levels at preoperative day( t = 3.34,3.53,3.44,3.69, all P ＜ 0. 05 ), the levels at the fourteenth day returned to normal levels,but the serum BGP showed a rising trend,the level was highest at the fourteenth day, and it was compared to the level at preoperative day( t = 2. 87, P ＜ 0. 05 ). Conclusion The changes of the serum inflammatory factor in the elder patients with femoral neck fracture during periopearative period showed regular pattern, and it was meaningful for understanding the developing and prognosis.

Objective:To identify metastasis-associated genes in salivary gland adenoid cystic carcinoma(ACC).Methods:Salivary gland adenoid cytic carcinoma cell line ACC-2 and its highly metastatic ACC-M cells were used to screen the metastasis-related genes in ACC by microarray technology.Two fluorescent cDNA probes labeled with Cy3 and Cy5 dyes,were prepared from the mRNA samples of ACC-2 and ACC-M cells by reverse transcription method.The two color probes were then mixed and hybridized on the cDNA chip constructed by double dots of 1152 human genes,and scanned at two wave lengths.Differentialy expressed genes of the two cell lines were analyzed using computer.Then seven of the differently expressed genes were further validated by RT-PCR technique.Results:Of the 1,152 known genes and expressed sequence tags,26 showed significantly different expression level(minimum 2 fold) between the two cell lines.Among the 26 genes,19 were up regulated(with ratio more than 2) and 7 were down(with ratio less than 1/2).The results of RT-PCR analysis for 7 differently expressed genes were coincident with those of microarray assay.Conclusions:Down regulation of LIFR,LCP1,DPEP1 and ABLIM1,and up regulation of DCC,MMP1 and CNTN2 may be related to the highly metastatic potential of ACC-M cell line.

Objective To observe the sedation and analgesia in surface anesthesia in conscious glossopha-ryngeal LMA for the clinical effect of laparoscopic hysterectomy.Methods 90 patients undergoing elective laparo-scopic total hysterectomy were randomly divided into three groups,30 cases in each group.The observation group (group A):sedation,analgesia,full surface anesthesia,control group 1 (B group)control group 2 (group C)were treated with endotracheal intubation under general anesthesia.At intubation and pull tube stage,the patients'reaction, hemodynamic changes,pneumoperitoneum 1 h blood gas and perioperative complications were observed.Results The mean arterial pressure(MAP)and heart rate(HR)of A group were (92.7 ±10.6)mmHg and (82.8 ±12.1)/min. Those of B group were (98.4 ±11.6)mmHg,(89.1 ±11.4)/min,C group were (111.2 ±12.5)mmHg,(104.1 ± 13.2)/min,those in A group and B group were better than C group(A group and C group,t =6.18,6.52,P <0.01;B group and C group,t =4.11,4.71,all P <0.01).The pulse oxygen saturation(SpO2 )and peak airway pressure (Paw)of A group were (99.6 ±0.4)and (17.3 ±2.1)mmHg,those in group B were (99.5 ±0.5)% and (17.6 ± 2.0)mmHg,group C were (99.5 ±0.5)% and (22.5 ±2.8)mmHg.The differences between A group and B group were statistically significant(compared with C group,t =14.3,8.14,all P <0.01;B group and C group,t =12.7, 7.78,all P <0.01).The incidence rates of perioperative complications and adverse reactions of A,B,C groups were 27%,33%,67%,that in C group was significantly higher than A group and B group(χ2 =9.64,6.67,all P <0.01). Conclusion The laryngeal mask airway was used for laparoscopic total hysterectomy under the condition of sedation and analgesia,and it can shorten the time of the whole body,prevent the difficulty of intubation,emergency airway and anesthesia related complications.

Objective:To synthesize autoinducer-2 by the clone and prokaryotic expression of Streptococcus mutans(S.mutans)UAl59 luxS gene and to observe the influence factors.Methods:The expression vector pET21 a(+)-luxS of S.mutans UAl59 was transformed into Escheriehia coli BL2l(DE3).The S-ribosylhomocysteinase(Luxs)expression was induced by IPTG.The His tag fusion protein was isolated by Ni-chelating column and identified by Western blotting.Finally the protein was renatured by dialysis method.S-ribosylhomo-cysteine (SAH)was catalyzed by s-adenosylhomocysteine nucleosidas (Pfs)and LuxS,and then AI-2 was syntheszed.The AI-2 activi-ty was examined by luminescence of Vibrio harveyi BB1 70 when the concentration of LuxS protein or pH(4 -1 2)or the concentration of sodium fluoride was changed in reaction mixes of AI-2 synthesis.Results:Compared with the control group,with the increase of LuxS protein concentration,the relative activity of in vitro synthesized AI-2 increased gradually(P <0.001 ).When pH was between 6 -1 0, the relative activity of AI-2 were the highest,beyond the range of pH,the relative activity of AI-2 decreased(P <0.001 ).When a final concentration of sodium fluoride was more than 0.3%,the luminescence values decreased(P <0.05).Conclusion:LuxS fusion protein can promote the production of AI-2.Optimum pH for AI-2 biosynthesis in vitro must be between 6-1 0.Biosynthesis of AI-2 is inhibited by sodium fluoride with final concentration of more than 0.3%.