1.Cloning of human uracil N-glycosylase and its detection in cancer tissues by quantitative RT-PCR.
Hong-Bo BAO ; Chuan-Bao ZHANG ; Jin-Fang WANG ; Chuan-Nong ZHOU ; Fang LIU ; Xiao-Hang ZHAO ; Shi-Jun QIAN
Chinese Journal of Biotechnology 2003;19(5):561-565
The uracil in DNA comes from either the misincorporation of dUTP in place of dTTP or deamination of cytosine. In the latter case, it can result in a GC to AT transition mutation if the uracil is not removed before DNA replication. Base excision repair (BER) is a major pathway for removing DNA lesions arising from endogenous processes as well as those induced by exposure to exogenous chemicals or irradiation. BER is initiated by DNA glycosylases that excise aberrant bases from DNA by cleavage of the N-glycosidic bond linking to the base of its deoxyribose sugar. Uracil N-glycosylase (UNG) is the enzyme responsible for the first step in the BER pathway that specifically removes uracil from DNA. The UNG gene undergoes both temporal and spatial regulation mainly at the level of transcription. Normally cancer cells undergo over-proliferation and up-regulate their UNG during tumorigenesis. In this study we examine the correlation between UNG level and carcinogenesis, and explore the possibility of using UNG as a marker for cancer diagnosis. Human UNG gene was amplified from the total RNA of the human choriocarcinoma cell line, JEG-3, by RT-PCR. After purification, the 942bp full-length UNG cDNA coding sequence was digested with EcoR I and Sal I, and cloned into the digested pET-21 to construct a recombinant vector, pUNG. The UNG protein was expressed under the control of T7 promoter in E. coli BL21 (DE3) cells induced with IPTG. After ultrasonic treatment, the cell lysate and precipitate were analyzed by SDS-PAGE and a 39kD band was detected. The plasmid was serially diluted at appropriate concentrations and employed as standards in the subsequent quantification. Total RNAs were extracted from 18 pairs of clinical samples, each pair contains a sample of esophageal squamous cell carcinoma (ESCC) tissue and its surrounding normal esophageal epithelia. The copy numbers of UNG mRNA in these RNA samples were determined by real-time quantitative RT-PCR using a Lightcycler (Roche). UNG was present in 13 cases of ESCC (13/18, n = 18) but absent in all of the normal tissues. The results indicated that there was a correlation between high level of UNG expression and the carcinogenesis of ESCC.
Carcinoma, Squamous Cell
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genetics
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metabolism
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Cell Line, Tumor
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Cloning, Molecular
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Electrophoresis, Polyacrylamide Gel
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Esophageal Neoplasms
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genetics
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metabolism
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Humans
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In Vitro Techniques
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Reverse Transcriptase Polymerase Chain Reaction
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Uracil-DNA Glycosidase
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genetics
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metabolism
2.Risk factors for oncologic outcome after surgical treatment in patients with gastric cancer: a multivariable analysis of 1031 patients.
Xiao-bao LI ; Hong-wei ZHANG ; Qing-chuan ZHAO ; Guang-long DONG ; Wei-zhong WANG
Chinese Journal of Gastrointestinal Surgery 2010;13(2):115-118
OBJECTIVETo investigate the risk factors for the prognosis in patients with gastric cancer undergoing surgery.
METHODSClinical data of 1031 cases who underwent gastric cancer resection from January 2003 to December 2007 were studied using univariable analysis and multivariable regression analysis.
RESULTSIn 1031 cases,95 (9.2%) cases were early-stage gastric cancer. The other 936 (90.8%) cases were advanced gastric cancer. The tumor was resectable in 980 (95.1%) cases, of which 874 (84.8%) were curative resection,106 (10.3%) were palliative, and 51 (4.9%) were bypass procedures or laparotomy alone. The stage-specific 5-year survival rates were 93.2% (stage IA), 65.1%(stage IB), 52.3% (stage II), 41.4% (stage IIIA), 16.6% (stage IIIB) and 10.6% (stage IV), respectively. The 1-, 3- and 5-year survival rates were 80.2%, 58.0% and 48.2%, respectively. The independent risk factors associated with the prognosis of these patients were tumor size, serum albumin, curative resection, TNM staging and multidisciplinary treatment in both univariable and multivariable analyses.
CONCLUSIONSEarly curative resection is the most important treatment for the patients with gastric cancer. Individualized surgical procedure combined with multidisciplinary treatment can improve the outcome. Tumor size, serum albumin level and TNM staging are important predictors of survival in patients with gastric cancer.
Adult ; Aged ; Aged, 80 and over ; Causality ; Female ; Gastrectomy ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Stomach Neoplasms ; epidemiology ; mortality ; surgery ; Survival Rate ; Treatment Outcome
3.Clinical application of real time-polymerase chain reaction in determining cytomegalovirus viral DNA load in renal transplant recipients.
Chuan-Bao ZHANG ; Hui-Ying LAI ; Hong-Tao XU ; Da-Guang WANG ; Fei XIAO
Chinese Medical Journal 2012;125(19):3575-3577
BACKGROUNDCytomegalovirus (CMV) remains a significant clinical problem among immunosuppressed renal transplant patients. Quantitative PCR assays have become the most common methods in the determination of CMV infections in transplant patients. This study was to determine the relationship between CMV infection and the acute rejection of the transplanted kidney.
METHODSPlasma samples from 77 renal transplant patients that were pre-transplant negative for CMV infection were tested using real-time quantitative PCR and CMV gene-specific primers. The detected viral loads were retrospectively compared with the acute rejection rate and the chronic or mild rejection rates of the renal transplant.
RESULTSCMV-DNA was detected in 29 of 77 recipients, yielding a positive rate of detection of 37.7% for this procedure. Twelve of the 21 recipients (57.1%) who suffered acute rejection had positive CMV-DNA. Among the 56 recipients suffered from chronic or mild rejection, 17 (30.4%) had positive CMV-DNA plasma. Moreover, of the 29 recipients who had detectable CMV-DNA after transplant, 12 (41.4%) suffered from acute rejection; of the 48 recipients with undetectable CMV-DNA, only nine (18.8%) developed acute rejection. Post-transplant patients with acute rejection had a higher rate (57.1% vs. 30.4%, P = 0.03) of post-transplant CMV infection than those with chronic or mild rejection.
CONCLUSIONCMV infection is a risk factor of acute renal transplant rejection and CMV infection should be prevented and treated in renal transplant recipients.
Adult ; Cytomegalovirus Infections ; diagnosis ; genetics ; DNA, Viral ; genetics ; Female ; Humans ; Kidney Transplantation ; adverse effects ; Male ; Middle Aged ; Real-Time Polymerase Chain Reaction ; methods ; Viral Load ; Young Adult
4.Diagnosis and treatment of testicular and epididymal diseases in Tibetan area.
Bao-Peng ZHANG ; Ping-Cuo GESANG ; Chuan-Hong LI
National Journal of Andrology 2012;18(4):353-355
OBJECTIVETo explore the clinical types, prevention and treatment of testicular and epididymal diseases in the Tibetan area.
METHODSWe retrospectively analyzed the clinical data of 105 cases of testicular or epididymal diseases treated in our department from 2007 to 2009. All the patients were permanent inhabitants in Tibet.
RESULTSAmong the 105 patients, the main types of testicular and epididymal diseases were tuberculosis (27 cases) and tumor (21 cases). And 99% of the patients were Tibetan farmers and herdsmen.
CONCLUSIONTibetan farmers and herdsmen have a poor knowledge about testicular and epididymal diseases and their prevention and treatment. Clinicians and related institutions need to strengthen health education on these diseases in Tibetan area to improve the early awareness, early diagnosis, early prevention and early treatment of testicular and epididymal diseases.
Adult ; Epididymis ; pathology ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Testicular Diseases ; diagnosis ; epidemiology ; therapy ; Testis ; pathology ; Tibet ; epidemiology
5.Endoscopic papillary balloon dilatation vs. endoscopic sphincteropapillotomy for common bile duct stones: a meta analysis.
Liang HE ; Xiao-ping GENG ; Hong-chuan ZHAO ; Da-chen ZHOU ; Fu-bao LIU ; Yi-jun ZHAO ; Guo-bin WANG ; Zhi-gong ZHANG ; Fan HUANG
Chinese Journal of Surgery 2013;51(6):556-561
OBJECTIVETo evaluate the safety and efficacy between endoscopic papillary balloon dilatation (EPBD) and endoscopic sphincteropapillotomy ( EST) for common bile duct stones using meta-analysis method.
METHODSRandomizd controlled trials comparing EPBD with EST for common bile duct stones and published from January 1990 to July 2012 were recruited. This meta-analysis was conducted to estimate short-term and long-term complications. Fixed random effect model or random effect model was established to analyze the data.
RESULTSTwelve randomizd controlled trials were included in this analysis. These studies included 1865 patients, 925 of them were treated with EPBD and 940 were treated with EST. The analysis of basic characteristics of these included studies showed that: compared to EST, patients in the EPBD group were younger (OR = -1.16, 95% CI: -1.49 to -0.84, P = 0.00), while in two groups, there were no significant difference (P > 0.05) in gender proportion, average size of stones, number of gallstones, previous cholecystectomy, the number of merged duodenal diverticulum, common bile duct diameter, the total follow-up time. Also, compared to EST, the overall stone clearance in the EPBD group was lower (OR = 0.64, 95% CI: 0.42 to 0.96, P = 0.03), pancreatitis incidence was higher (OR = 2.67, 95% CI: 1.61 to 4.43, P = 0.00), incidence of bleeding (OR = 0.12, 95% CI: 0.04 to 0.34, P = 0.00), acute cholecystitis (OR= 0.39, 95% CI: 0.18 to 0.84, P = 0.02), total long-term complication rate (OR = 0.53, 95% CI: 0.36 to 0.77, P = 0.01), stone recurrence rate more than a year were lower (OR= 0.48, 95% CI: 0.26 to 0.90, P = 0.02). While in two groups, there were no significant difference (P > 0.05) in the stone removal on 1 '' attempt, the total near-term complications and acute cholangitis.
CONCLUSIONSOn the basis of lower rates of bleeding, EPBD seems to be preferred strategy over EST for endoscopic remove of common bile duct stones in patients who have coagulopathy. Although stone recurrence rate more than a year of EPBD is lower, but the overall stone clearance rate is lower and the risk of pancreatitis is higher than that of EST.
Dilatation ; Gallstones ; surgery ; Humans ; Postoperative Complications ; epidemiology ; Randomized Controlled Trials as Topic ; Sphincterotomy, Endoscopic ; Treatment Outcome
6.Determination of serum cholesterol by isotope dilution gas chromatography mass spectrometry
Jiang-Tao ZHANG ; Cui-Hua HU ; Chuan-Bao ZHANG ; Hai-Jian ZHAO ; Rong MA ; Jie-Hong XIE ; Jing WANG ; Jun DONG ; Shu WANG ; Ziyu SHEN ; Wenxiang CHEN ;
Chinese Journal of Laboratory Medicine 2001;0(03):-
Objective To develop a new isotope dilution gas chromatography mass spectrometry method (ID/GC/MS) for the measurement of serum cholesterol.Methods Serum was mixed with an isotope labeled internal standard ([3,4-~(13)C]-cholesterol) and treated with alcoholic sodium hydroxide to hydrolyze cholesterol ester to cholesterol.Cholesterol and internal standard was extracted and derived by N, O-Bis(trimethylsilyl) trifluoroacetamide to trimethylsilyl ethers.The derivation products were analyzed by capillary column GC combined with electron impact MS using scan and selected ion monitor (SIM) modes. Signals of cholesterol internal standard were corrected for the contributions from cholesterol and the signal ratio of cholesterol to internal standard for the calibrators were linearly regressed against cholesterol concentrations.The resulted regression equation was used for the calculation of serum cholesterol concentrations.Results The new ID/GC/MS method showed a mean within-run coefficient variance (CV) of 0.04%-0.81%.Comparison with two levels of standard reference material (SRM1951a) of National Institute of Standards and Technology (NIST) displayed a bias of 0.19% and 0.90% respectively.Conclusion A time-gaining ID/MS method has been established that is highly precise and accurate and can be used for the measurement of serum cholesterol.
7.CT-Guided Microcoil Localization of Small Peripheral Pulmonary Nodules to Direct Video-Assisted Thoracoscopic Resection without the Aid of Intraoperative Fluoroscopy
Zhen-guo HUANG ; Cun-li WANG ; Hong-liang SUN ; Chuan-dong LI ; Bao-xiang GAO ; He CHEN ; Min-xing YANG
Korean Journal of Radiology 2021;22(7):1124-1131
Objective:
To evaluate the feasibility, safety, and effectiveness of CT-guided microcoil localization of solitary pulmonary nodules (SPNs) for guiding video-assisted thoracoscopic surgery (VATS).
Materials and Methods:
Between June 2016 and October 2019, 454 consecutive patients with 501 SPNs who received CTguided microcoil localization before VATS in our institution were enrolled. The diameter of the nodules was 0.93 ± 0.49 cm, and the shortest distance from the nodules to the pleura was 1.41 ± 0.95 cm. The distal end of the microcoil was placed less than 1 cm away from the nodule, and the proximal end was placed outside the visceral pleura. VATS was performed under the guidance of implanted microcoils without the aid of intraoperative fluoroscopy.
Results:
All 501 nodules were marked with microcoils. The time required for microcoil localization was 12.8 ± 5.2 minutes. Microcoil localization-related complications occurred in 179 cases (39.4%). None of the complications required treatment. A total of 463 nodules were successfully resected under the guidance of implanted microcoils. VATS revealed 38 patients with dislocated microcoils, of which 28 underwent wedge resection (21 cases under the guidance of the bleeding points of pleural puncture, 7 cases through palpation), 5 underwent direct lobectomy, and the remaining 5 underwent a conversion to thoracotomy. In 4 cases, a portion of the microcoil remained in the lung parenchyma.
Conclusion
CT-guided microcoil localization of SPNs is safe and reliable. Marking the nodule and pleura simultaneously with microcoils can effectively guide the resection of SPNs using VATS without the aid of intraoperative fluoroscopy.
8.CT-Guided Microcoil Localization of Small Peripheral Pulmonary Nodules to Direct Video-Assisted Thoracoscopic Resection without the Aid of Intraoperative Fluoroscopy
Zhen-guo HUANG ; Cun-li WANG ; Hong-liang SUN ; Chuan-dong LI ; Bao-xiang GAO ; He CHEN ; Min-xing YANG
Korean Journal of Radiology 2021;22(7):1124-1131
Objective:
To evaluate the feasibility, safety, and effectiveness of CT-guided microcoil localization of solitary pulmonary nodules (SPNs) for guiding video-assisted thoracoscopic surgery (VATS).
Materials and Methods:
Between June 2016 and October 2019, 454 consecutive patients with 501 SPNs who received CTguided microcoil localization before VATS in our institution were enrolled. The diameter of the nodules was 0.93 ± 0.49 cm, and the shortest distance from the nodules to the pleura was 1.41 ± 0.95 cm. The distal end of the microcoil was placed less than 1 cm away from the nodule, and the proximal end was placed outside the visceral pleura. VATS was performed under the guidance of implanted microcoils without the aid of intraoperative fluoroscopy.
Results:
All 501 nodules were marked with microcoils. The time required for microcoil localization was 12.8 ± 5.2 minutes. Microcoil localization-related complications occurred in 179 cases (39.4%). None of the complications required treatment. A total of 463 nodules were successfully resected under the guidance of implanted microcoils. VATS revealed 38 patients with dislocated microcoils, of which 28 underwent wedge resection (21 cases under the guidance of the bleeding points of pleural puncture, 7 cases through palpation), 5 underwent direct lobectomy, and the remaining 5 underwent a conversion to thoracotomy. In 4 cases, a portion of the microcoil remained in the lung parenchyma.
Conclusion
CT-guided microcoil localization of SPNs is safe and reliable. Marking the nodule and pleura simultaneously with microcoils can effectively guide the resection of SPNs using VATS without the aid of intraoperative fluoroscopy.
9.Experimental study on two-way application of traditional Chinese medicines capable of promoting blood circulation and removing blood stasis with neutral property in cold and hot blood stasis syndrome I.
Er-Wei HAO ; Jia-Gang DENG ; Zheng-Cai DU ; Ke YAN ; Zuo-Wen ZHENG ; Qin WANG ; Li-Zhen HUANG ; Chuan-Hong BAO ; Xiu-Qiong DENG ; Xiao-Yan LU ; Zhi-Ling TANG
China Journal of Chinese Materia Medica 2012;37(21):3302-3306
OBJECTIVETo study the action characteristics of "two-way application and conditioned dominance" of traditional Chinese medicines with neutral property by observing the action characteristic of 10 traditional Chinese medicines capable of promoting blood circulation and removing blood stasis with neutral property in the microcirculation in rats with heat stagnation and blood stasis syndrome.
METHODThe rat model with heat stagnation and blood stasis syndrome was established by injecting carrageenan and dry yeast, and the rat model with cold stagnation and blood stasis syndrome was built by the body freezing method. Ten traditional Chinese medicines with neutral property, including 5 with hot property and 5 with cold property, were selected for intervention to observe blood flow rate and flow state indicators in rat auricles and make a comparative analysis on action characteristics of traditional Chinese medicines with neutral property.
RESULTANOVA showed that among the 10 traditional Chinese medicines with neutral property, 6 such as Typhae Pollen, Sappan Lignum and Vaccariae Semen can obviously increase the blood flow rate (P < 0.01 or P < 0.05) in the above two models; all of the 5 traditional Chinese medicines with cold property can increase the blood flow rate (P < 0.01 or P < 0.05) in the rat model with heat stagnation and blood stasis syndrome, but only Salvia miltiorrhiza can increase the blood flow rate (P < 0.01 or P < 0.05) in the rat models with cold stagnation and blood stasis syndrome, while other medicines showed no notable effect; among the 5 traditional Chinese medicines with hot property, Carthamus tinctorius and Ligusticum chuanxiong can increase the blood flow rate (P < 0.01 or P < 0.05) in the rat models with cold stagnation and blood stasis syndrome, but had no obvious effect to the blood flow rate in the rat models with heat stagnation and blood stasis syndrome. According to the analysis on average blood flow rate, traditional Chinese medicines with natural and cold properties showed similar effect on heat stagnation and blood stasis syndrome and better effect in increasing blood flow rate than those with hot property; those with natural and hot properties showed similar effect and better effect in increasing blood flow rate than those with cold property.
CONCLUSIONUnder the condition of heat stagnation and blood stasis syndrome, traditional Chinese medicines with neutral property have the similar action characteristics with those with cold property; wile under the condition of cold stagnation and blood stasis syndrome, traditional Chinese medicines with neutral property have the similar action characteristics with the Chinese medicinal herbs with hot property. This indicates the action characteristics of "two-way application and conditioned dominance" of traditional Chinese medicines with neutral property to some extent.
Animals ; Blood Circulation ; drug effects ; Blood Coagulation ; drug effects ; Male ; Medicine, Chinese Traditional ; Microcirculation ; drug effects ; Rats ; Syndrome
10.A case-control study on drinking green tea and decreasing risk of cancers in the alimentary canal among cigarette smokers and alcohol drinkers.
Li-na MU ; Xue-fu ZHOU ; Bao-guo DING ; Ru-hong WANG ; Zuo-feng ZHANG ; Chuan-wei CHEN ; Guo-rong WEI ; Xiao-ming ZHOU ; Qing-wu JIANG ; Shun-zhang YU
Chinese Journal of Epidemiology 2003;24(3):192-195
OBJECTIVETo explore the role of green tea in decreasing the risks of gastric cancer, liver cancer, esophageal cancer among alcohol drinkers or cigarette smokers.
METHODSA population based case-control study was conducted in Taixing, Jiangsu province.
RESULTSIn Taixing city, identified cases of stomach, liver and esophageal cancers were chosen with informed consent. The numbers were 206, 204, 218 respectively. Controls were chosen from normal population having lived in the area for longer than 10 years, also with informed consent. Green tea drinking seemed to have decreased 81%, 78%, 39% risk for the development of gastric cancer, liver cancer and esophageal cancer among alcohol drinkers. It might also have decreased 16%, 43%, 31% on the risks of developing the three kinds of cancers among cigarette smokers. Interaction assessment showed that drinking green tea could significantly decrease the risk of gastric cancer and liver cancer among alcohol drinkers, with ORs of interaction item 0.23 (95% CI: 0.10 - 0.55) and 0.25 (95% CI: 0.11 - 0.57) respectively.
CONCLUSIONHabit of drinking green tea seemed to have significant protective effects on the development of both gastric and liver cancer among alcohol drinkers while, green tea also having some protective effect on esophageal cancer among alcohol drinkers and on three kinds of cancers among cigarette smokers.
Adult ; Aged ; Alcohol Drinking ; adverse effects ; Case-Control Studies ; China ; epidemiology ; Digestive System Neoplasms ; epidemiology ; etiology ; prevention & control ; Esophageal Neoplasms ; etiology ; Female ; Flavonoids ; administration & dosage ; Humans ; Liver Neoplasms ; epidemiology ; etiology ; prevention & control ; Male ; Middle Aged ; Phenols ; administration & dosage ; Polyphenols ; Risk ; Smoking ; adverse effects ; Stomach Neoplasms ; epidemiology ; etiology ; prevention & control ; Tea ; chemistry