1.The effects of 5-Aza-2-deoxycytidine, a methylation inhibitor, on the proliferation of androgen-sensitive prostate cancer cell line, and on the regulation of methylation of glutathione s-transferase P1 and retinoic acid receptorβ2 gene
Mo SHEN ; Binghua CHEN ; Ping ZHOU ; Wu ZHOU ; Zhihua TAO
Chinese Journal of Geriatrics 2014;33(3):306-310
Objective To investigate the effects of 5-Aza-2'-deoxycytidine (5-Aza-dc),a methylation inhibitor,on the proliferation of androgen-sensitive prostate cancer line (LNCaP),and on its regulation of methylation on glutathione s transferaseP1 (GSTP1) and retinoic acid receptorβ2 (RARβ2) gene.Methods LNCaP cells were treated with 5-Aza-dc in different concentration,CCK8 method was used to detect the growth of LNCaP cells.The methylation of GSTP1 and RARβ2 gene in LNCaP cell was detected by nested methylation specific polymerase chain reaction (nMSP).Results The proliferation of LNCaP cells was inhibited after exposed to 5-Aza-dc.The methylation of GSTP1 and RARβ2 gene was changed from hypermethylation to demethylation by the 5-Aza-dc.These effects were dose-and time-dependent within certain concentration of 5-Aza-dc,but LNCaP cells grew better after 72 h than within 48 h when exposed to 5 Aza dc below 1.0 μmol/L.Also the methylation of GSTP1 and RARβ2 gene changed from hypermethylation to demethylation by the 5-Aza-dc was not different when exposed to 5-Aza-dc below 1.0 μmol/L within 72 h and 48 h.Conclusions 5-Aza-dc may effectively inhibit the growth of LNCaP cells and reverse the DNA methylation damage in some tumor suppressor genes,but the continuity and stability of low dose 5-Aza-dc is changeable.The study will provide a research basis for clinical treatment.
2.Immune surveillance on postoperative patients with advanced gastric cancer treated with concurrent chemotherapy and dendritic cells vaccine
Guoqiang Lü ; Binghua XU ; Xiaoming SHEN ; Li LI ; Fang XIANG
Chinese Journal of Digestion 2009;29(12):825-828
Objective To evaluate clinical outcome in postoperative patients with advanced gastric cancer treated with concurrent chemotherapy and dendritic cells (DCs) vaccine as well as alteration of immune function. Methods Sixty-four postoperative patients with advance gastric cancer were divided into control group and DC vaccine group, In control group, 38 patients were treated with LCH regimen consisted of 5-fluorouracil 0.75 g/d_(1-5) and oxaliplatin 0.2 g/d_1. In DC vaccine group,blood sample was obtained from 26 patients who were followed by LCH regimen treatment next day.One week after the chemotherapy, patients were immunized with DC vaccine for 2 times at interval of one week. The second cycle was performed after 28 days. The percentages of T lymphocytes and natural killer (NK) cells and cytokine levels before and after treatment were compared between two groups. The therapeutic effects (including no remote metastasis and enlarged lymph nodes in cavity and/or tumor reduced in volume) were also evaluated. Results The concentrations of T lymphocytes (CD3~+ and CD4~+ ), NK cells, interleukin (IL)-2, IL-12 and interferon-γ in DC vaccine group were significantly increased after vaccination compared with those before vaccination (P<0.05), and even higher than those in control group (P<0.05). The effective rate was higher in DC vaccine group (80. 76%) than that in control group (68.42%) with significant difference (P<0. 05). The side effects of chemotherapy such as the decreased peripheral white blood ceils and immune cells were less serious in DC vaccine group compared with control group (P< 0.05), while the uncomfortable incidence of gastrointestinal tract and peripheral neuritis showed no significant difference between two groups (P>0.05). Conclusions Application of concurrent chemotherapy and DC vaccine in patients with advanced gastric cancer after surgical treatment may achieve a short-term efficacy, meanwhile it can reduce the side effects induced by chemotherapy.
3.Effects of dexmedetomidine-propofol-fentanyl combined anesthesia on somatosensory and motor evoked potentials in patients undergoing cervical spine surgery
Sheng LIN ; Shuqin NI ; Dongxiu SUN ; Wei SHAO ; Binghua SHEN
Chinese Journal of Anesthesiology 2010;30(11):1284-1286
Objective To evaluate the effects of dexmedetomidine-propofol-fentanyl combined anesthesia on somatosensory evoked potentials (SEPs) and motor evoked potentials (MEPs) in patients undergoing cervical spine surgery. Methods Thirty-six patients undergoing cervical spine surgery were randomly divided into 2 groups (n = 18 each): propofol-fentanyl combined anesthesia group (group C) and dexmedetomidine-propofol-fentanyl combined anesthesia group (group D). Anesthesia was induced with TCI of propofol and iv injection of fentanyl.After the consciousness disappeared, a laryngeal mask airway was placed and the patients were ventilated. In group D, dexmedetomidine 0.5 μg/kg was injected over 10 min after the consciousness disappeared, followed by an infusion at a rate of 0.5 μg·kg-1 ·h-1 until the end of surgery. In group C, the equal volume of normal saline was administered instead of dexmedetomidine. SEPs (P15-N20) amplitudes and latency were measured and recorded before dexmedetomidine administration and at 10 min of dexmedetomidine infusion. The no-elicitation of MEPs was recorded. Results Compared with group C, there was no significant difference in P15-N20 amplitudes and latency in group D. The no-elicitation rate of MEPs in two groups was 0. Conclusion Dexmedetomidine-propofol-fentanyl combined anesthesia does not affect SEPs and MEPs in patients undergoing cervical spine surgery.
4.Expression of THANK in human PBMC activated with different stimulators
Dong WU ; Feng SHEN ; Yonghua LOU ; Binghua JIAO ; Mengchao WU
Journal of Cellular and Molecular Immunology 2001;17(4):304-306
Aim To analyze THANK gene expression in peripheral blood mononuclear cells(PBMC) stimulated with different stimulators and to clone whole length human THANK gene. Methods PBMC were conventionally isolated and cultured in RPMI1640 containing 10% FCS. After stimulated with LPS,TNF α ,IL 2,IFN γ ,PHA or PMA,the THANK gene expression in PBMCs was analyzed by RT PCR and THANK cDNA was cloned. Result RT PCR detection showed that THNAK gene expressed in PBMCs after stimulated with interferon γ for 3 days, whereas THANK'expression could not be detected after stimulated with LPS,TNF α ,IL 2,IFN γ ,PHA or PMA respectively. Then THANK gene was cloned by cloning PCR product and sequenced. Conclusion Human THANK gene is cloned successfully, thus providing the possibility for further research of THANK'function.
5.Recurrence prevention by embedding 5-FU microparticles on the cutting surface after hepatectomy for HCC
Tao ZHENG ; Feng XIE ; Jiamei YANG ; Weifeng SHEN ; Li GENG ; Binghua DAI ; Rongxi SHEN
Chinese Journal of General Surgery 2012;27(10):805-807
Objective To evaluate embedding 5-FU microparticles during operation after hepatectomy for small hepatocellular carcinoma to prevent recurrence. Methods A retrospective analysis on 184 patients was done.All patients received liver resection for HCC from 2007 Mar.to 2008 Mar.In 90 cases,5-FU releasing microparticles were embeded on liver cutting surface after the tumor was resected (group A).94 cases in which no micropaticles were given served as controls (group B).All patients had a single tumor( ≤5 cm) without visible vessel invasion. Results The postoperative disease-free survival ( DFS),and median period of DFS of group A vs.group B was 43 months vs 35 months.Postoperative DFS rates of 1-,2-and 3-year were 86%,76% % and 64% respectively in A group,while they were 77%,61%and 49% respectively in B group ( x2 =4.77,P < 0.05 ).In two groups the liver and kidney main function index( TB,ALB,ALT,BUN,Scr)on day 7 and postoperative complications were not statistically different (P > 0.05 ). Conclusions Embeding 5-FU microparticle after hepatectomy for HCC could increase the postoperative DFS rates,and patients do not suffer from significant liver and kidney malfunctions.
6.Expression, denaturation, renaturation and purification of THANK protein
Dong WU ; Feng SHEN ; Yonghua LOU ; Min PENG ; Binghua JIAO ; Mengchao WU
Academic Journal of Second Military Medical University 2001;0(09):-
Objective: To prepare highly purified THANK protein. Methods: THANK was efficiently expressed in E.coli as inclusion bodies. After bacteria were lysed under ultrasonication, TE buffer,1% TritonX and 2 mol /L urea was used to efficiently extract inclusion bodies. After denaturation with 8 mol/L urea,THANK was partially purified by Sephacryl S-200 chromatography, and then subsequent refolding step was optimized for a maximal recovery of biological active protein. The renatured THANK was purified to homogeneity with Q Sephadex Fast Flow gel filtration and then desalted by Sephadex G-25 chromatography. Results: The purity of biologically active THANK was above 97% in SDS-PAGE densitometric studies. Conclusion: An effective method of denaturation, renaturation and purification of recombinant THANK is established.
7.Treatment of old osteoporotic vertebral compression fractures with Sky bone expander percutane-ous kyphoplasty
Binghua SHEN ; Wei DU ; Jianqing LIU ; Shiqiao LU ; Jun LIANG ; Chuanbao WANG ; Leisheng WANG
Chinese Journal of Trauma 2009;25(3):227-231
Objective To discuss the clinical effect of Sky bone expander percutaneous ky-pbeplasty (Sky-PKP) in treatment of old osteoperotic vertebral compression fractures. Methods The study involved 27 patients (27 vertebrae) with old osteoporotic vertebral compression fractures treated by Sky-PKP from March 2005 to June 2007. Normotopia, lateral and dynamia radiographs, CT scanning and MRI were performed preoperatively to verify fluid collection in the vertebral body, vacuum phenomenon and open-close phenomenon. Visual analog scale (VAS), Oswestry disability index (ODI), anterior body height, middle line body height, posterior body height and kyphotic angle changes were measured on a lateral radiograph before and after treatment. Results All patients were followed up for mean 6.2 months, which showed no severe complications. VAS score was decresed from preoperative 7.8 to postop-erative 3.1 and ODI from 65% to 37%. However, The anterior vertebral height and middle line vertebral height were recovered for 4.6 nun and 5.7 mm respectively compared with preoperation. Correction of ky-photic angle was mean 5.6°postoperatively. There was no sitatistical changes in regard to posterior body height before and after operation. Conclusions Sky-PKP is a reasonable procedure for treatment of old osteoporotic vertebral compression fractures under strict control of indications, especially with vacuum phenomenon, open-close phenomenon and fluid collection. While high degree of difficulty in puncturation results in insignificant correction of kyphotic angle and body height.
8.The clone and expression of human THANK gene
Dong WU ; Feng SHEN ; Yonghua LOU ; Yuping ZHU ; Binghua JIAO ; Mengchao WU ;
Academic Journal of Second Military Medical University 2000;0(07):-
Objective: To clone THANK gene and express its extracelluar fragment. Methods: Using RNA isolated from HL 60 cell lines, THANK cDNA was amplified by RT PCR. The fragment was linked to pMD18 T vector and sequenced, and then the extracellular fragment of THANK was subcloned into pET vector and THANK protein expression was induced.Results: A 858 bp DNA fragment was amplified and the cDNA sequence was identical with the published sequence encoding THANK gene. Western blot showed that THANK protein with a relative molecular weight of 2.6?10 4 was expressed. Conclusion: Human THANK gene was cloned and expressed successfully, which provides a base of further research of THANK gene. [
9.Application of terminal ileum suspension in laparoscopic operation for low rectal cancer
Hui ZHAO ; Yifan SHI ; Zenghui YANG ; Chuanqing BAO ; Xiaoming SHEN ; Binghua XU
The Journal of Practical Medicine 2017;33(22):3741-3744
Objective To explore the value of terminal ileum suspension in the treatment of low rectal cancer.Methods 80 patients with low rectal cancer who underwent laparoscopic operation in our hospital from June 2015 to February 2017,were randomly divided into two groups:the control group (group C) and the test group (group T),40 cases in each group.In group C,laparoscopic radical resection of rectal cancer (Dixon) was performed and in group T Dixon was combined with terminal ileum suspension.Peripheral blood nutritional indicators (total plasma protein,albumin,pre-albumin,transferrin) and major electrolytes of two groups were observed 1 day before operation and 1,3 and 7 days after operation.The two groups were compared in terms of first exhaust time,postoperative hospital stay,total costs for hospitalization,postoperative discomforts and complications.Results There were no statistical differences in the levels of nutrition indicators and electrolytes between them (P > 0.05) and neither it was with first exhaust time,hospital stay,total costs of hospitalization,incidence of postoperative discomforts and complications (P > 0.05).The re-operation rate of group T with anastomotic leak was significantly lower than group C (P < 0.05).Conclusion Terminal ileal suspension does not affect patients' postoperative recovery without increasing the patient's suffering and economic burden,and can effectively reduce the reoperation rate caused by anastomotic leak.It is easy to operate.
10.Quantitative evaluation of extracellular volume fraction after acute ST segment elevation myocardial infarction by iodine density based on spectral detector CT
Binghua CHEN ; Dongaolei AN ; Jie HE ; Rui WU ; Ruoyang SHI ; Chongwen WU ; Ting YUE ; Ziyang FAN ; Yisi DAI ; Jialu SHEN ; Yingying DING ; Ji WANG ; Jun PU ; Xingbiao CHEN ; Lianming WU ; Jianrong XU
Chinese Journal of Radiology 2020;54(6):527-533
Objective:To explore the feasibility of quantitative evaluation of extracellular volume (ECV) fraction in acute ST-segment elevation myocardial infarction (STEMI) by dual-layer spectral detector CT.Methods:The clinical and imaging data of 20 patients with STEMI who underwent cardiac contrast-enhanced CT and MRI from January to October 2019 in Renji Hospital affiliated to Shanghai Jiaotong University School of Medicine were retrospectively analyzed.The dual spectral detector was used in the enhanced CT scan of the coronary artery with retrospectively gate and the late iodine enhancement with prospective gate. Conventional image and holographic spectral image were obtained by iterative and spectral reconstruction. The short axis image of the heart matched with MR image was obtained by multiplanar reconstruction. Based on the data of spectral based image, the IDD map was reconstructed for the calculation of myocardial CT-ECV during the late iodine enhancement. ECV of infarcted myocardium, salvageable myocardium and remote myocardium based on CT and MRI were calculated respectively. Bland-Altman consistency test and intra group correlation coefficient analysis (ICC) were used to compare the consistency of two measurements and different methods. The correlation between CT-ECV and MRI-ECV was compared by Spearman method.Results:The CT-ECV values of infarcted, salvageable, and remote myocardium were 51.21 (49.27, 53)%, 38.64 (36.17, 40)%, and 51.21 (49.27, 53)%, respectively. The difference was statistically significant ( H= 43.17, P<0.01). The CT-ECV value of infarcted myocardium was significantly higher than that of salvageable myocardium and remote myocardium ( Z=-24.60, 35.40, P<0.01), but there was no significant difference between salvageable myocardium and remote myocardium ( Z= 10.80, P=0.15). The T 1 values of infarcted myocardium, salvageable myocardium and remote myocardium were (1 554.85±70.94), (1 443.85±67.28) and (1 307.05±91.73) ms respectively, the difference was statistically significant ( F=51.35, P<0.01). The T 1 value of infarcted myocardium was higher than that of salvageable myocardium and remote myocardium ( t=-5.07, 9.55, P<0.01), and salvageable myocardium was significantly higher than that of remote myocardium ( t=5.38, P<0.01). The MRI-ECV values of infarcted myocardium, salvageable myocardium and remote myocardium were 55.00 (49.27, 57.75)%, 33.50 (29.00, 35.00)%,and 27.00 (26.00, 29.00)%, respectively. The difference was statistically significant ( Z= 47.12, P<0.01). MRI-ECV of infarcted myocardium was significantly higher than that of salvageable myocardium and remote myocardium ( Z=37.45, -20.30, P< 0.01), and salvageable myocardium was significantly higher than that of remote myocardium ( Z = 17.15, P<0.05). The difference between CT-ECV and MRI-ECV measured by two physicians was good. The bias of Bland-Altman analysis was -0.1% (95% CI:-5.5%-5.2%), 0.8% (95% CI:-9.8%-8.2%), and the ICC values were 0.92 and 0.94, respectively. The bias of Bland-Altman analysis in CT-ECV and MRI-ECV consistency test was 4.00% (95% CI:-9.0%-16.9%) and ICC value was 0.88, which had a good correlation ( r=0.75, P=0.001). Conclusions:The iodine density based ECV fromdual-layer spectral detector CT can be used to quantitatively evaluate the changes of extracellular space after acute STEMI, which is helpful to quantitatively evaluate the histological changes after myocardial ischemia.