1.Applications of Phospho-histone H2AX Enzyme-Linked Immunosorbent Assay in Vitro Genotoxicity Evaluation of Cigarette Smoke
Liwei FU ; Huan CHEN ; Jin YANG ; Hongwei HOU ; Qingyuan HU
Chinese Journal of Analytical Chemistry 2014;(3):320-325
Phospho-histone H2AX(γH2AX) has been widely used in vitro genotoxicity evaluation of ionizing radiation, carcinogenic substances and cigarette smoke as an important biomarker of DNA double stranded breaks ( DSBs) . The study developed an enzyme-linked immunesorbent assay for detection of the content ofγH2AX in cells to evaluate the genotoxicity of cigarette smoke. The study exposed CHO cells with cigarette smoke total particulate matter ( TPM) and cigarette smoke condensate ( CSC) of different dose, dose-effect relationship between smoke exposure and DNA damage and the poisonous difference of cigarette smoke components has been investigated by detecting the content change ofγH2 AX in cells under different exposure time and dose. In addition, reactive oxygen species ( ROS) in cells were also detected to study the mechanism of cigarette smoke exposure induced DSBs. The experiment result showed that the required time for the level ofγH2AX in cells grew to the peak prolonged with increased exposure dose. Once the level of γH2AX in cells grew to the peak value, then it decreased slowly; the level of γH2AX in cells rised with increasing dose of cigarette smoke;The effect of TPM on the content change of γH2AX was more than CSC; Moreover, smoke exposure could induce concentration increase of ROS in cells, and a good correlation of content change of ROS and γH2AX in cells were found, free radicals in cigarette smoke may be one major cause of DSBs.
2.Clinical effect of total excision of the thyroid and most resection in the treatment of thyroid nodule, and its influence to thyroid function
Huasen HOU ; Yuequan FU ; Weina JIN ; Maoqiang LI
Chinese Journal of Primary Medicine and Pharmacy 2016;23(11):1660-1663
Objective To investigate the safety of thyroid gland resection and primary resection in the treatment of thyroid nodule and its influence to thyroid function.Methods 86 patients with thyroid nodules were selected as the research subjects.The patients were randomly divided into two groups.43 cases in the observation group implemented the thyroid gland resection treatment.43 cases in the control group received thyroid gland subtotal resection.The curative effect,safety,thyroid function and other indicators were compared between the two groups.Results The operation time and hospitalization time in the observation group after operation[(105.65 ± 12.54)min;(6.35 ± 2.01)d]were shorter than those in the control group[(149.41 ± 13.68)min;(9.62 ± 2.45)d].The amount of bleeding during operation in the control group [(134.51 ± 9.64) mL] was significantly higher than the observation group [(84.62 ± 6.35) mL],there was significant difference between the two groups (P < 0.05).The total effective rate of the observation group was 95.55%,which was significantly higher than 72.09% in the control group,the difference was statistically significant (P < 0.05).Before operation,the between the two groups had no significant difference (P > 0.05).After surgical treatment,serum FT3 and serum FT4 levels in two groups were decreased,but those in the observation group[(11.62 ± 3.02),(51.24 ± 7.25)pmol/L] were significantly lower than the control group [(14.14 ± 5.11) pmoL/L;(60.52 ± 6.35) pmol/L],there were obvious differences (P < 0.05).The incidence rate of complications such as throat edema,hemorrhage,postoperative hoarseness in the observation group was 9.30%,which in the control group was 23.26%,the difference was statistically significant (P < 0.05).Conclusion The thyroid gland resection therapy for thyroid nodules has high safety and significant curative effect,it can quickly improve the thyroid function,reduce relapse rate,has higher application value in benign and malignant tumors indistinguishable after surgery.It can reduce the length of stay in hospital,is conducive to the recovery of patients,it is worthy of clinical promotion.
3.An analysis of the result of tumor screening for health examination participants
Zhiming ZHAO ; Jun FU ; Guichun YU ; Huiwen JIN ; Yanwei HOU ; Li TAO
Chinese Journal of Health Management 2011;05(3):143-145
Objective To analysis the result of tumor screening for health examination participants.Methods Tumor screening was applied for 15 863 health examination participants without malignant tumor disease history.Clinical examination,laboratory tests and imaging studies were comprehensively analysed combining the results of other medical items.The cancer detection rate and asymptomatic rate were compared among groups of different age,gender and existence of relevant clinical symptoms,respectively.χ2 test and Fisher exact test were adopted for statistical analysis.Results Among 475 tumor patients with a definite diagnosis,116 were malignant tumor;The total detection rate of the group who were younger than 35 year-old(0.617%)was higher than other groups,the detection rate of intracranial neoplasms of the two groups who were younger than 45 year-old (0.206% and 0.132%,respectively)was higher than other groups,the total detection rate of 75-85 group (0.248%)was lower than other groups; The detection rate of lung cancer,liver cancer of the male(0.228% and 0.080%,respectively)was higher than the female,The detection rate of breast cancer and intracranial neoplasms of the female(0.366% and 0.100%,respectively)was higher than the male; There are 67 patients without correlated clinical symptoms,the asymptomatic rate was 57.8%.Conclusions Tumor screening was of importance in health management,which should be carried out actively in health examination.
4.XAGE-1b mRNA expression in the blood specimens of patients with liver cancer, cirrhosis and benign liver diseases and in healthy individuals
Zhenyu HOU ; Zeya PAN ; Jingzhong ZHANG ; Tongming JIN ; Siyuan FU ; Weiping ZHOU
Chinese Journal of Hepatobiliary Surgery 2011;17(8):645-647
Objective To investigate the significance of XAGE-1bmRNA expression in the blood specimens of patients with primary liver cancer, cirrhosis and benign liver diseases and in healthy individuals. Methods Venous blood specimens of patients with primary liver cancer (n= 125), cirrhosis (n= 23), benign liver diseases (n= 34) and healthy individuals (n = 41 ) were collected. XAGE-1b mRNA was detected by real-time fluorescence quantitative PCR. Results The expression levels of XAGE-1b mRNA in patients with primary liver cancer, cirrhosis, benign liver diseases and healthy in dividuals were 3.72 (0.93, 10.2) ×10-5, 0 (0, 0. 56) ×10-5, 0 (0, 0)×10-5, 0 (0, 0) ×10-5, respectively. The XAGE-1b mRNA expression in patients with primary liver cancer was obviously higher than the patients with cirrhosis, benign liver diseases and in healthy individuals. The expression levels for patients with cirrhosis was higher than patients with benign liver diseases and in healthy individuals. The expression levels for patients with benign liver diseases and healthy individuals were similar. With a optimal cut-off value of 8. 385 × 10-7 , the sensitivity, specificity, positive predictive value, and negative predictive value of XAGE-lb mRNA for diagnosing primary liver cancer were 80. 0%, 89.8%, 90.9% and 77.9% respectively. The positive rates for patients with primary liver cancer and cirrhosis were 80.0% and 30.4% respectively. Conclusion XAGE-lb mRNA can be used as a tumor marker for primary liver cancer. It contributes to the differentiation between primary liver cancer, cirrhosis and benign liver diseases.
5.Preparation of bovine serum albumin nanoparticles loaded with gemcitabine and the cytotoxic effect on pancreatic cancer cells in vitro
Jinming LI ; Wei CHEN ; Jiang LONG ; Chen JIN ; Weiyue LU ; Quanxing NI ; Deliang FU ; Huimin HOU
Chinese Journal of Pancreatology 2008;8(5):295-297
Objectives To investigate preparation of gemcitabine albumin nanoparticles, and its property of slow-release, the cytotoxic effect on pancreatic cancer cells (PANC1) in vitro, for improving the effect of regional intra-arterial infusion chemotherapy in pancreatic cancer with new medicament in the future. Methods The gemcitabine albumin nanoparticles were prepared with bovine serum albumin and gemcitabine with the desolvation-crosslink method, the concentration of gemcitabine was detected by high performance liquid chromatography (HPLC). The cytotoxic effect on pancreatic cancer cells in vitro were detected with MTT colorimetric assay. Results The mean diameter of gemcitabine albumin nanoparticles was (156.2±2.2) nm, and Zeta potential was (-20.4±1.41)mV, drug loading was 10.8%, drug release time in virto was 3 hours respectively. Gemcitabine albumin nanoparticles (0.01~50 μg/ml) had a 31%~44% inhibitory rate on PANC1 cell, which was similar to the inhibitory rate of same concentration of gemcitabine (26%~47%). Conclusions The new preparation of gemcitabine albumin nanoparticles had obvious drug slow-release effect, which may help improve the effect of regional intra-arterial infusion chemotherapy for pancreatic cancer.
6.Bioinformatics analysis and construction of eukaryotic expression plasmid of Cx50 V64G mutation
Ping, LIU ; Ying, LIN ; Yue-Ying, YANG ; Jian-Qiu, ZHENG ; Ying, HOU ; Di, JIN ; Xiao-Bo, FU ; Hong-Mei, MA
International Eye Science 2007;7(5):1206-1208
AIM : To construct and analyze eukaryotic expression plasmid inserted by Cx50 with V64G mutation through bioinformatics software.METHODS: The full coding domain sequence of Cx50 with V64G mutation was acquired from the blood of patients with cataract and was cloned into pcDNA3.1 /Amp (+).The constructed plasmid was identified with PCR , enzyme digestion and sequencing. The analysis of Cx50 with V64G mutation was performed with bioinformatics software.RESULTS : Cx50 with V64G mutation was successfully amplified and its eukaryotic expression plasmid was constructed. Valine-64 is well conserved in the first extracellular loop of connexin 50 in different species and also in different human α -type gap junctional proteins.CONCLUSION : The successive reconstruction and verification of eukaryotic expression plasmid containing Cx50 with V64G mutation established the foundation for further studying the mechanism of cataract.
7.Efficacy of interferon-alpha therapy for HBeAg-negative chronic hepatitis B and its influencing factors.
Qian-guo MAO ; Kang-xiang LUO ; Qun-fang FU ; Xiao-rong FENG ; Ya-bing GUO ; You-fu ZHU ; Jie PENG ; Jin-jin HOU
Chinese Journal of Hepatology 2004;12(10):582-584
OBJECTIVETo investigate the efficacy of interferon-alpha (IFN-alpha) therapy for HBeAg-negative chronic hepatitis B.
METHODSSixty-five Chinese HBeAg-negative chronic hepatitis B patients were treated with 5 MU recombinant rIFN-alpha 1b subcutaneously thrice weekly for 5 to 24 months, followed by 12 months of treatment-free follow-up; one hundred and eighty-eight Chinese HBeAg-positive patients served as controls. For each patient, serum alanine transaminase (ALT) was measured biochemically and serum HBV DNA level was detected with fluorescent-quantitative PCR, HBeAg with enzymoimmunoassay every 1 to 3 months during therapy and during the follow-up period. HBeAg loss (only for HBeAg-positive cases), HBV DNA undetectable, and ALT normalization: the three together were considered a combined response.
RESULTSRates of combined response were similar in HBeAg-negative patients (58.5%, 38/65) or HBeAg-positive ones at the end of treatment (weighted chi square test, chi2 = 1.878, P<0.05), but were higher at the end of the follow-up period in the HBeAg-negative cases (75.4%, 49/65) (weighted chi square test, chi2 = 4.796, P<0.05). Furthermore, relapse rates at the end of the follow-up period, were also similar in HBeAg-negative patients (15.8%, 6/38) or HBeAg positive (chi2 = 0.205, P>0.05). Combined response was achieved at a median of 6.0 months (2-16 months) of treatment course in HBeAg-negative patients while at a median of 6.0 months (1-22 months) in HBeAg-positive cases (Z = -0.186, P>0.05, by the Wilcoxon rank sum test). The only factor predictive of combined response, by binary logistic regression analysis, was inflammatory activity in the liver biopsy. Gender, age, baseline ALT level, baseline HBV DNA level, and anti-HBe were not predictive factors.
CONCLUSIONInterferon-alpha therapy induces a similar primary and sustained response in HBeAg-negative and in HBeAg-positive chronic hepatitis B patients.
Female ; Follow-Up Studies ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; immunology ; therapy ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Treatment Outcome
8.Relationship between single nucleotide polymorphisms in IL28B gene and response to interferon treatment in chronic hepatitis B patients.
Chu-ming CHEN ; Fu-yuan ZHOU ; Yuan-ping ZHOU ; Yan-li ZENG ; Wei LI ; Jin-lin HOU
Journal of Southern Medical University 2011;31(12):2012-2015
OBJECTIVETo explore the relationship between the single nucleotide polymorphisms (SNPs) of rs12979860 and rs8099917 in IL28B gene and the response to interferon treatment in hepatitis B e antigen (HBeAg)-positive patients with chronic hepatitis B patients.
METHODSPeripheral blood samples were collected from 82 HBeAg-positive patients with chronic hepatitis B receiving interferon treatment, including 38 with favorable response to the treatment and 44 without response. IL28B gene was amplified from the chromosomal DNA, and rs8099917 SNP was genotyped based on PCR-RLFP and rs12979860 SNP by sequencing.
RESULTSIn the responsive patients, the distribution frequencies of TT and TG+GG genotypes and allele G in SNPrs8099917 were 81.6% (31/38), 18.4% (7/38), and 9.2% (7/76), as compared to the frequencies of 97.7% (43/44), 2.3% (1/44), and 1.1% (1/88) in nonresponsive patients, respectively. The frequencies showed significant differences between the responsive and nonresponsive patients (P=0.014 for genotypes and P=0.025 for allele G). The distribution frequencies of CT genotypes and allele T in SNPrs12979860 showed no differences between the responsive and nonresponsive patients (P<0.05).
CONCLUSIONrs8099917 SNP is probably associated with the response to interferon treatment in HBeAg-positive patients with chronic hepatitis B, and Allele G may be predictive of the treatment success.
Adult ; Alleles ; Antiviral Agents ; therapeutic use ; Female ; Genotype ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; genetics ; therapy ; Humans ; Interferons ; therapeutic use ; Interleukins ; genetics ; Male ; Polymorphism, Single Nucleotide ; Young Adult
10.Factors related to chronic hepatitis B relapse after interferon-alpha treatment: a follow-up study.
Ding-Li LIU ; Kang-Xian LUO ; Xiao-Rong FENG ; Qun-Xiang FU ; Jin-Lin HOU
Journal of Southern Medical University 2007;27(8):1264-1270
OBJECTIVETo investigate the related to relapse of chronic hepatitis B (CHB) after recombinant interferon-alpha (rIFN-alpha) treatment.
METHODSThis investigation involved 523 pathologically confirmed CHB patients including 403 HBeAg-positive and 120 HBeAg-negative patients, who were treated with 5 MU rIFN-alpha subcutaneously thrice a week for 6-25 months. For each patient, serum alanine aminotransferase (ALT) was measured biochemically, serum HBV DNA level detected with quantitative fluorescent PCR, and HBeAg level with enzyme immuoassay every 1-3 months during therapy and every 3-6 months during the follow-up period.
RESULTSEarly response to rIFN-alpha treatment was observed in 302 (57.7%) patients at the end of treatment, among whom 39.4% (119/302) suffered relapse during the follow-up for 39.2-/+21.5 months. Age, HBeAg status before treatment, and follow-up duration were the predictive factors for post-treatment relapse. The mean age of patients with CHB relapse was significantly higher than that of the sustained responders (P<0.001), and the relapse rates in HBeAg-negative group (55.8%, 43/77) were significantly higher than that in HBeAg-positive group (33.8%, 76/225) at the end of follow up (P<0.001). The relapse rate and accumulative relapse rates at each year during the follow-up (for 5 years as the longest) differed significantly (P<0.001, P=0.000), but the accumulative relapse rates differed little between the years after the initial 2 of the follow-up (P=0.670). The relapse was not related to the patient's gender, pretreatment serum ALT, HBV DNA, grade of liver inflammation, stage of liver fibrosis, or duration of treatment. In HBeAg-positive patients, however, the mean HBV DNA was significantly higher in relapse group than in sustained response group (P=0.017).
CONCLUSIONAge, pretreatment HBeAg status, and follow-up duration are independent predictive factors for post-treatment CHB relapse. In HBeAg positive patients, pretreatment serum HBV DNA is also one of the risk factors for relapse.
Adult ; Age Factors ; Alanine Transaminase ; blood ; DNA, Viral ; blood ; Female ; Follow-Up Studies ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; drug therapy ; therapy ; Humans ; Interferon-alpha ; therapeutic use ; Logistic Models ; Male ; Recurrence ; Treatment Outcome