1.Basic and clinical study of interferon a in hepatocellular carcinoma.
Hui-chuan SUN ; Lu WANG ; Wei-zhong WU
Chinese Journal of Surgery 2008;46(21):1606-1608
3.The unusual typing of short tandem repeats.
De-jian LU ; Yu-chuan CHEN ; Hui-ling LU
Journal of Forensic Medicine 2002;18(2):118-123
Short tandem repeats (STRs) have been widely used in forensic sciences such as stain analysis and paternity testing. Although most of STR typing could give the reliable and clear results, some unusual typing have been observed in forensic practice. The anomalous typing could result from a lot of causes, including DNA genetic variation, poor quality or quantity of DNA template, different typing system or method, nonspecific reaction in PCR or anomalous electrophoresis migration. The unusual results may disturb the right interpretation of STR typing.
DNA Fingerprinting/methods*
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Forensic Medicine/methods*
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Gene Frequency
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Genotype
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Humans
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Polymerase Chain Reaction/methods*
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Polymorphism, Genetic
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Tandem Repeat Sequences/genetics*
4.Detection of coxsackie virus B3 gene in myocardium and spleen samples in myocarditis with sudden death.
Yu Chuan CHEN ; Jian Ding CHENG ; Hui Hui MA ; Bing Jie HU ; Chun Lan YAO
Journal of Forensic Medicine 2001;17(2):74-76
OBJECTIVE:
To detect the Coxsackie virus B3(CVB3) gene in myocardium and spleen tissues in viral myocarditis(VMC) with sudden death and to explore the diagnostic method for VMC by means of seeking pathogene.
METHODS:
By in situ RT-PCR, the detection of CVB3 gene in myocardium and spleen sections were performed in sudden death group caused by VMC and non-cardiac death group.
RESULTS:
In VMC group, CVB3 gene-positive signals were seen in myocardium sections(3 out of total 8 cases, No. 1, 4, 7 cases) and spleen sections(4 out of total 8 cases, No. 2, 4, 6, 7 cases). In non-cardiac death group, no positive signals were detected in both myocardium and spleen tissues.
CONCLUSION
Positive detection of CVB3 gene in both myocardium and spleen maybe an important character of VMC and can improve the detecting pathogene in diagnosing VMC.
Death, Sudden
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Enterovirus B, Human/genetics*
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Heart/virology*
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Humans
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Myocarditis/virology*
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Reverse Transcriptase Polymerase Chain Reaction
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Spleen/virology*
5.Effects of ARNT2 gene on invasion and migration of human hepatocellular carcinoma HCCLM6 cell line.
Wei-Wei LI ; Wei-Zhong WU ; Ying LIANG ; Chuan-Li XIAO ; Zhong-Hua TAO ; Lu WANG ; Hui-Chuan SUN ; Jia FAN
Chinese Journal of Hepatology 2010;18(1):27-31
OBJECTIVETo investigate the effects of ARNT2 on invasion and migration of HCCLM6 cells.
METHODSFour short hairpin oligos targeting to ARNT2 were s cloned into the pLVTHM vector. Lentiviral vectors shRNA-ARNT2i, pCMV-dR8.74 and pMD2G were cotransfected into 293T cells using Lipofectamine 2000. HCCLM6 was infected with virus supernatant. ARNT2 mRNA and protein expressions were detected using quantitative Real time-PCR and Western blot, respectively. The invasion and migration of HCCLM6 cells were evaluated using wound healing assay and cell invasion assay in vitro. Statistical analysis was performed with SPSS 16.0.
RESULTSThe relative mRNA levels of ARNT2 were 0.154+/-0.024, 0.860+/-0.145, 1.004+/-0.009 in shRNA-ARNT2i virus infected HCCLM6 cells, mock-infected cells and control vector virus infected cells (F = 113.14, P more than 0.01). The expression of ARNT2 at protein level was 16.45+/-1.6, 44.56+/-2.07 in the HCCLM6 cells infected with shRNA-ARNT2i virus and negative control vector virus, respectively (t = 18.58, P less than 0.01). The scrape wound of HCCLM6 cells infected with shRNA-ARNT2i virus healed faster than cells infected with control vector virus or mock-infected cells. The number of cells invading through Matrigel was higher in the HCCLM6 cells infected with shRNA-ARNT2i virus (13.25+/-1.04) than that in mock-infected HCCLM6 cells and the HCCLM6 cells infected with negative control vector virus (6.50+/-2.56, 6.75+/-2.05) (F = 29.645, P less than 0.01).
CONCLUSIONInhibition of ARNT2 gene promotes the invasion and migration of HCCLM6 cells.
Aryl Hydrocarbon Receptor Nuclear Translocator ; genetics ; metabolism ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; metabolism ; Blotting, Western ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; genetics ; Humans ; Lentivirus ; genetics ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Neoplasm Invasiveness ; Polymerase Chain Reaction ; methods ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Transfection
6.Expression of augmenter of liver regeneration in hepatic tumor cells and its clinical significance.
Hang SUN ; Hui-feng YU ; Chuan-xin WU ; Xiao-qin GUAN ; Qi LIU
Chinese Journal of Hepatology 2005;13(3):205-208
OBJECTIVETo investigate the effects of augmenter of liver regeneration (ALR) on the proliferation of hepatocytes and hepatic tumor cells and the expression of ALR in herpatocellular carcinoma (HCC).
METHODSPrimary rat hepatocytes, QGY and HepG2 cells were cultured separately with ALR from different species. Cell proliferation was detected by their 3H-TdR uptake. The expression of ALR was examined in 9 normal hepatic tissues and 21 HCC cases using immunohistochemistry method.
RESULTSDifferent ALRs could stimulate the proliferation of HepG2 and QGY cells in a dose-dependent way in vitro, but all ALR had no influence in the proliferation of primary rat hepatocytes. The expression of ALR was absent in normal hepatic tissues, but present in all HCC hepatic tissues. However, the expression of ALR had no relationship with the differentiation and size of the carcinomas.
CONCLUSIONALR might play an important role in the occurrence and development of HCC.
Animals ; Carcinoma, Hepatocellular ; metabolism ; Hepatocytes ; metabolism ; Liver Neoplasms ; metabolism ; Liver Regeneration ; drug effects ; physiology ; Male ; Proteins ; genetics ; metabolism ; Rats ; Rats, Wistar
7.Construction and identification of expressing siRNA plasmid against human augmenter of liver regeneration.
Lin TANG ; Qi LIU ; Hang SUN ; Ni TANG ; Hui GUO ; Jian-Chuan DENG
Chinese Journal of Hepatology 2004;12(9):534-537
OBJECTIVESTo detect whether there is an expression of human augmenter of liver regeneration (hALR) in HepG2 cells. To develop a kind of RNAi that specifically targets human augmenter of liver regeneration by synthesizing small interfering RNA (siRNA) in vivo, and to assess the inhibitory effect of this siRNA on hALR expression.
METHODSThe expression of hALR in HepG2 cells was observed with immunocytochemistry. The RNAi plasmid pSIALR-A and the unrelated control plasmid pSIALR-B were transfected into HepG2 cells. Forty-eight hours after transfection, the protein level of hALR was measured with immunocytochemistry; meanwhile, the reverse transcription PCR (RT-PCR) was performed to detect the expression of hALR mRNA.
RESULTShALR was expressed by HepG2 cells. siRNA plasmid pSIALR-A, which targets the cDNA of hALR and the unrelated control plasmid pSIALR-B, was successfully constructed. Both immunocytochemistry and RT-PCR showed that pSIALR-A inhibited the expression of hALR in HepG2 cells significantly, compared with that of pSIALR-B.
CONCLUSIONThe results showed that the small interfering RNA targeting hALR suppresses the expression of hALR in a sequence-specific manner
Base Sequence ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Molecular Sequence Data ; Plasmids ; genetics ; Proteins ; genetics ; pharmacology ; RNA, Small Interfering ; genetics ; Transfection
8.A study on the relationship between glutathione S-transferases gene polymorphism and susceptibility response to hypoxia.
Hui-qin YAN ; Xue-chuan SUN ; Kong-xiang LIU ; Sheng-wei WANG ; Tao LIU
Chinese Journal of Applied Physiology 2006;22(3):334-337
AIMTo investigate the relationship between glutathione S-transferases gene polymorphism and susceptibility response to hypoxia.
METHODSIn the case-control study, the gene polymorphisms of glutathione S-transferases were tested in Tibetan mountaineers and sea-level Han Chinese by multiple-PCR and PCR-RELP.
RESULTSThe frequency of GSTT1 null genotype was significant different between Tibetan mountaineers and sea-level Han Chinese (P < 0.05), OR = 1.86 (95% CI = 1.01-3.39), and also for GSTP(1-105) mutant genotype in two groups (P < 0.01), OR = 2.19 (95% CI = 1.16-4.13). There was significant difference between A allele and G allele of GSTP(1-105) groups (P < 0.01). There was no difference for GSTM1 null genotype between two groups (P > 0.05), OR = 0.78 (95% CI = 0.43 - 1.42).
CONCLUSIONGSTT1 and GSTP(1-105) genotype may be associated with susceptibility response to altitude hypoxia.
Adult ; Alleles ; China ; Genotype ; Glutathione S-Transferase pi ; genetics ; Glutathione Transferase ; genetics ; Humans ; Hypoxia ; genetics ; Male ; Mountaineering ; Polymorphism, Genetic ; Reactive Oxygen Species ; metabolism ; Young Adult
9.Reconstructing the JAK/STATs signal pathway restored the anti-proliferative response of MHCC97 on interferon alpha.
Wei-zhong WU ; Hui-chuan SUN ; Yan-qin GAO ; Lu WANG ; Zhao-you TANG ; Kang-da LIU
Chinese Journal of Hepatology 2006;14(4):277-280
OBJECTIVETo elucidate the roles of JAK/STATs signal pathway on anti-proliferative effects induced by IFN-alpha in MHCC97.
METHODSAn IRF9 expression vector was transfected into MHCC97 with Dosper. The expression of IRF9, cycle regulating proteins and the forming of ISGF3 complex were detected using Western blot and EMSA, respectively. Cell proliferation and distribution were monitored using MTT and flow cytometry.
RESULTSHigh expression of IRF9 restored the anti-proliferative response of MHCC97 on IFN-alpha treatment and delayed the cell transition from S phase to G2 phase induced by IFN-alpha.
CONCLUSIONThe integrity and functions of JAK/STATs signal pathway played an important role in mediating the anti-proliferative effects of IFN-alpha in MHCC97.
Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Interferon-Stimulated Gene Factor 3, gamma Subunit ; genetics ; Interferon-alpha ; metabolism ; pharmacology ; Janus Kinases ; genetics ; physiology ; Liver Neoplasms ; genetics ; metabolism ; pathology ; STAT Transcription Factors ; genetics ; physiology ; Signal Transduction ; Transfection
10.Transcription factor activity profile of human hepatocellular carcinoma cell lines with different metastatic potentials.
Qi PAN ; Lu WANG ; Hui-chuan SUN ; Yin-kun LIU ; Sheng-long YE ; Zhao-you TANG
Chinese Journal of Hepatology 2006;14(1):37-40
OBJECTIVESTo examine the activities of transcription factors (TFs) in human hepatocellular carcinoma (HCC) cell lines with different metastatic potentials, so as to identify the TFs associated with HCC metastasis.
METHODSTranscription factor activity profile of Hep3B, MHCC97L and MHCC97H, three HCC cell lines with different metastatic potentials, were examined using protein/DNA array. Electrophoretic mobility shift assays (EMSA) and Western blot were used to confirm the results obtained by protein/DNA array.
RESULTSFrom a total of 345 screened TFs, 7 activity differential TFs were found, of which 5 showed increased activity, including p53, hypoxia inducible factor-1 alpha (HIF-1alpha), signal transducer and activator of transcription 3 (Stat3) and Sp1, and 2 showed decreased activity including Rb and Smad3.
CONCLUSIONThe abnormal functioning of transcription factors is closely associated with HCC metastasis. Our present findings could be of help in expanding our understanding of the mechanism of HCC metastasis and identify new predictive biomarkers and therapeutic targets.
Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; DNA Fingerprinting ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; Protein Array Analysis ; Transcription Factors ; classification ; genetics ; metabolism