1.The immunoprecipitation method for the determination of ?-glutamyltransferase complexed to two lipoproteins in serum and its application for the diagnosis of liver malignancies
Jiaping YU ; Aihua WANG ; Pengfei GU ; Jun CHEN
Chinese Journal of Laboratory Medicine 2000;0(06):-
Objective To establish the immunoprecipitation method to detect the activity of ?-glutamyltransferase (GGT) complexed to low-density-lipoprotein and very-low-density-lipoprotein in serum (LDL-LDL-GGT) and to observe the diagnostic efficiency in discriminating liver malignancies from other hepatopathy diseases.Methods Using apo B antibody as a precipitant to precipitate the parts of GGT which connected to LDL and VLDL. 65 cases of liver malignancies, 53 cases of cirrhosis, 32 of chronic hepatitis and 75 of health people were analysed. Results Apo B antibody can precipitate LDL and VLDL thoroughly with various serum samples. The withinrun CV are 3 6%~8 2% and the betweenrun CV are 5 5%~9 8%. The linearity range is 0~587 U/L. With the cut off value of 10 U/L, the sensitivity for liver malignancies is 86 2%, the specificities for discriminating liver malignancies from cirrhosis and chronic hepatitis is 67 9% and 81 3% respectively. Conclusion It has important significance for the diagnosis of liver malignancies with the immunoprecipitation method for determination of serum LDL-VLDL-GGT activity.
2.Prokaryotic expression of human ubiquitous mitochondrial creatine kinase, preparation of its antiserum and clinical application
Jian ZHANG ; Aihua WANG ; Pengfei GU ; Jiaping YU
Academic Journal of Second Military Medical University 1999;0(12):-
Objective:To clone the full-length cDNA of human ubiquitious mitochondrial creatine kinase(uMtCK) and express its protien in E. coli. The protein was used to immunize rabbit to prepare anti-uMTCR polyclonal antibody which was then used to detect the expression of uMtck in gastrointestinal carcinoma. Methods: A 1 062 bp segement was amplified from human HaLa cell by RT-PCR,which was then inserted into pMD18-T plasmid and proved to be the coding sequence of uMtCK by endonuclease digestion and sequencing. The coding sequence of uMtCK was inserted into pQE30 plasmid and recombinant plasmid pQE30-uMtCK was transformed into E. coli. The expressed protein was purified with a His. Bind column and was analyzed with SDS-PAGE and Westen blot and the activity of uMtCK was tested. An antiserum against uMtCK was perpetrated by immunizing rabbit with purified uMtCK. The expression of uMtCK in 59 patients with gastric and colonic cancer was detected by immunohistochemistry. Results:The coding sequence of uMtCK was successfully obtained by RT-PCR and the product was successfully inserted into pQE30 to construct a recombinant vector pQE30-uMtCK. The expression of uMtCK was effective and soluble, A good specific antiserum against uMtCK was proved by Western blot. The expression of uMtCK of gastric and colonic cancer was 76. 5%. Conclusion:The coding sequence of uMtCK is coined and expressed in E.coli. uMtCK is a prospective marker for the diagnosis of gastric and colonic cancer.
3.Isolation of algicolous fungi and screening of their antitumor activity
Pengfei HAO ; Tianjiao ZHU ; Tiezheng JIA ; Qianqun GU ; Weiming ZHU
Chinese Journal of Marine Drugs 2001;0(05):-
Objective To isolate algicolous fungi from algae and obtain antitumor strains.Methods Selective isolation of algicolous fungi from algae was done,and antitumor activity corresponds to their lethiferous effect on brine shrimp and cytotoxicity against K562 cell was determined.Results 221 strains of algicolous fungi were isolated from 28 algae samples collected from intertidal zones of the Yellow Sea.Eight strains showed lethiferous activity and eight showed cytotoxicity.Conclusion The results indicated that algicolous fungi would be one of the important resources to search for new antitumor compounds.
4.Cytotoxicity of GS and IL-2 Activated PBMC to Malignant Glioma Cells
Junsheng LUO ; Lixue GU ; Huanjiu XI ; Bingjie WEI ; Xingbo LIU ; Jianwu QIU ; Pengfei ZHANG ; Hongren SHAN
Chinese Journal of Cancer Biotherapy 2000;7(4):273-274
Objective: To investigate a new method for improving the therapeutic effect on malignant glioma. Methods: A new type of killer cells, named GS-LAK, was induced by means of costimulating the peripheral ginsenoside(GS) and interleukin-2 (IL-2). Comparing with control group-LAK cells, cytotoxicity of GS-LAK cells against malignant glioma cells(BT325) was examined with MTI method. Results: It showed that GS-LAK cells exhibited some advantages over LAKcells in proliferation, cytotoxicity, as well as the utilizing of IL-2. Conclusion: The application of GS-LAK cells mightopen a new prospect to clinical therapeutic approach to malignant glioma.
5.Quantitative Determination of Agarotetrol in Agarwood by HPLC-DAD
Yufan GU ; Qian ZHANG ; Huixia HUO ; Zheng HUANG ; Jing ZHANG ; Yunfang ZHAO ; Jun LI ; Pengfei TU
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(12):2643-2646
This study was aimed to establish a quantitative method for the determination of agarotetrol in agarwood to control its quality. The HPLC analysis was performed on a Diamonsil C18 column (4.6 mm í 250 mm, 5 μm), eluted with a mobile phase of water with 0.1% formic acid and acetonitrile in gradient mode with the flow rate of 0.7 mL·min-1. The detection wavelength was set at 252 nm. The column temperature was 30oC. The results showed that the separation of agarotetrol and adjacent peaks were more than 1.5, which achieved a baseline separation. A good lin-earity (r = 0.999 8) was observed in the range of 2.0-125.0 μg·mL-1, with the average recovery of 102.75%. The a-garotetrol cannot be less than 0.15% based on the data of Agarwood samples. It was concluded that the method was accurate and reliable to determine the content of agarotetrol, which can be used for the quality control of agarwood.
6.Risk factors analysis of ulcerative colitis-associated colorectal cancer
Yanhui GU ; Guangsen HAN ; Shijia ZHANG ; Yuzhou ZHAO ; Jian LI ; Pengfei MA ; Yanghui CAO ; Mingke HUO
Chinese Journal of Digestive Surgery 2017;16(7):736-740
Objective To explore the risk factors of ulcerative colitis-associated colorectal cancer (UC-CRC).Methods The retrospective case-control study was conducted.The clinicopathological data of 536 patients with ulcerative colitis (UC) who were admitted to the Henan Tumor Hospital from March 2004 to June 2015 were collected.Observation indicators:(1) follow-up results:cases with follow-up,follow-up time,cases of UC-CRC,age of onset,pathological type of UC-CRC;(2) risk factors analysis affecting occurrence of UC-CRC:gender,age of onset,course of disease,severity of disease,disease classification,extent of lesion,smoking history,family history of colorectal cancer,anemia,hypoproteinemia,body weight loss,extraintestinal manifestations,colonic polyps,backwash ileitis,atypical hyperplasia,anxiety or depression,treatment method and regular endoscopy reexamination.Follow-up using outpatient examination and telephone interview was performed to detect prognosis of patients up to April 2017.Patients underwent colonoscopy once every 6 months within 3 years after diagnosis and once every 1 year after 3 years.Measurement data with skewed distribution were described as M (range).The univariate analysis was done using the chi-square test and Fisher exact probability.The multivariate analysis was done using the Logistic regression model.Results (1) Follow-up results:of 536 patients,450 were followed up for 26.0-120.0 months,with a median time of 76.4 months.During the follow-up,16 patients were complicated with UC-CRC,including 9 males and 7 females.Age of onset of colorectal cancer was 14-78 years,with an average age of onset of 44 years.Pathological type:high-differentiated right colon adenocarcinoma was detected in 5 patients,high-and moderate-differentiated left colon adenocarcinoma in 3 patients,left colon signetring cell carcinoma in 2 patients,moderate-differentiated rectal tubular adenocarcinoma in 3 patients,highdifferentiated rectal papillary adenocarcinoma in 2 patients and malignant lymphoma in 1 patient.(2) Risk factors analysis affecting occurrence of UC-CRC:the results of univariate analysis showed that course of disease,extent of lesion,colonic polyps and atypical hyperplasia were risk factors affecting occurrence of UC-CRC (x2 =14.848,18.885,10.554,P<0.05).The results of multivariate analysis showed that course of disease > 10 years,lesion involving the whole colon,colonic polyps and atypical hyperplasia were independent risk factors affecting occurrence of UC-CRC (OR=12.893,17.847,7.326,19.742,95% confidence interval:1.726-74.337,1.445-89.793,1.263-43.128,3.625-96.524,P<0.05).Conclusion The course of disease > 10 years,lesion involving the whole colon,atypical hyperplasia and colonic polyps are independent risk factors affecting occurrence of UC-CRC.
7.Irreversible electroporation for treatment of locally advanced pancreatic cancer
Junjun SU ; Ming SU ; Kai XU ; Pengfei WANG ; Shichun LU ; Wanqing GU ; Yongliang CHEN
Chinese Journal of Hepatobiliary Surgery 2017;23(7):464-467
Objective To evaluate the overall survival in patients with locally advanced pancreatic cancer (LAC) treated with irreversible electroporation (IRE) and chemotherapy.Methods A retrospective study on the overall survival of 30 patients with LAC treated with IRE,and 30 patients with LAC treated with chemotherapy from July 2015 to October 2016 in the PLA General Hospital was conducted.Results For the 30 patients with LAC who underwent IRE successfully,there were 21 women and 9 men.The median age was 59 (36 ~81) years.Twenty-four patients had primary pancreatic head cancer and 6 had body cancer.Twelve (40.0%) of these patients had chemotherapy after the IRE ablation.The 90-day mortality in the IRE treated patients was 3 (10.0%).For the 30 patients with LAC who were treated with chemotherapy,the 90-day mortality was 6 (20.0%).In comparison of the IRE treated patients with the chemotherapy treated patients,improvements on disease-free survival (6 months vs.4 months,P < 0.05) and overall survival (11 months vs.5.6 months,P < 0.05) were observed.Conclusion IRE ablation of LAC was safe and could potentially improve overall survival when compared with the standard chemotherapy treatment.
8.Laparoscopic extraperitoneal radical prostatectomy
Changjun YIN ; Wei ZHANG ; Wei ZHANG ; Min GU ; Qiang Lü ; Xiaoxin MENG ; Pengfei SHAO ; Zhengquan XU ; Yuangeng SUI
Chinese Journal of Urology 2010;31(3):199-202
Objective To present the experience of laparoscopic extraperitoneal radical prostatectomy and evaluate its safety and efficacy. Methods A total of 91 patients diagnosed with localized prostate carcinoma were admitted from February 2003 to June 2008. The level of serum PSA ranged from 7. 5 - 47. 0 ng/ml(mean 14. 0 ng/ml). The volume of the prostate ranged from 35 - 75 ml(mean 52 ml). Biopsy was performed before the operation and the pathological results revealed prostate carcinoma with Gleason score no more than 8. CT, MR and ECT revealed there was no lymph node or seminal vesicle involvement and there was no bone metastasis. The procedures were performed with an-tegrade techniques and pelvic lymphadenectomies were performed in 32 cases and nerve-sparings were performed in 11 cases. Results The operation duration ranged from 105 - 270 min (mean 173 min). Intraoperative blood loss was 110 - 1200 ml(mean 315 ml). Incontinence occurred in 19 cases in early stage and 18 cases recovered within 3 months. Positive surgical margin occurred in 11 cases. There was no complication of urethra stricture during 3 - 30 months' follow-up. No lymph node was involved in 32 cases with pelvic lymphadectomy. Five of the 11 cases received nerve-sparing prostatectomy had normal erectile function during the follow-up. Conclusions Laparoscopic extraperitoneal radical prostatectomy is a safe, effective and efficient surgical procedure with the minimal invasion, less morbidity and rapid recovery. Laparoscopic radical prostatectomy is emerging as an alternative to open radical prostatectomy.
9.Association between serum level of interleukin-18 and insulin resistance in women with polycystic ovary syndrome
Yifei ZHANG ; Yisheng YANG ; Jie HONG ; Weiqiong GU ; Chunfang SHEN ; Min XU ; Pengfei DU ; Xiaoying LI ; Guang NING
Chinese Journal of Endocrinology and Metabolism 2008;24(3):278-281
Objective To evaluate the relationship between interleukin-18(IL-18) and insulin resistance, measured by euglycemic hyperinsulinemic clamp, in women with polycystic ovary syndrome (PCOS). Methods Forty-two young women with PCOS and 38 age-and body mass index (BMI)- matched control women were recruited in this study. A complete hormonal assay was performed and serum IL-18 level was determined in each subject. And euglycemic hyperinsulinemic clamp test was completed in 41 of the above subjects. Results Serum IL-18 levels were increased in the PCOS women, as compared with the control (P=0.033). When all subjects were divided into lean and obese groups, the IL-18 levels were slightly increased in the obese subjects (P=0.902). IL-18 levels were negatively correlated with all the clamp parameters [mean glucose infusion rate (M), M-to-insulin ratio (M/I) and glucose metabolic clearance rate (MCRg), r =-0.419,-0.396,and-0.405,P=0.006,0.010 and 0.009 respectively], but were positively associated with HOMA-β index(r=0.334, P=O.035). Conclusion Serum IL-18 level was significantly increased in PCOS women and was strongly associated with the parameters obtained from the euglycemic hyperinsulinemic clamp, indicating that IL-18 may be an important mediator between inflammation and insulin resistance.
10.Lymph nodes sorting based on nanometer carbon staining and supplying artery distribution in right-sided colon carcinoma
Yuzhou ZHAO ; Guangsen HAN ; Xiaobing CHEN ; Ying LIU ; Jian LI ; Yanhui GU ; Pengfei MA ; Chenyu LIU ; Mingke HUO
Chinese Journal of General Surgery 2017;32(1):12-14
Objective To evaluate a novel lymph node (LN) sorting method on surgical resected sample guided by nanometer carbon staining and principle following the supplying artery tributory in rightsided colon carcinoma.Methods From May 2015 to June 2016,51 patients were randomly divided into two groups adopting traditional LN sorting method and that of a combination of nanometer carbon and artery guided.The final LN status were compared between the 2 groups.Results The total LN number and the positive LN in novel method group were higher than control group (437 vs.349,70 vs.54).The dissection time used,the number of harvested positive LN that was < 5 mm were significantly different [(13.1 ± 3.2) minvs.(17.8 ±3.8)min,t=4.75,P=0.000;1.0±l.0vs.0.2 ±0.6,t=3.51,P=0.000].The number of patients with harvested LN less than 12,the rate of positive lymph nodes,the rate of metastasis were not significantly different (all P > 0.05) between the two groups.Conclusions Use of nanometer carbon development combined with artery approach facilitates LN sorting,yielding more positive LNs,and increating the accuracy of pathological staging in right-sided colon cancer.