1.The Application of Heating Effect in Breeding of Microorganism
Xing-Qiang GAO ; Yun-Hong HUANG ; Fei DAI ; Xue-Qin FU ; Zhong-Er LONG ;
Microbiology 2008;0(10):-
Advances in mechanism and application of the heating effect in breeding of microorganism are reviewed in this paper. Heat produces mutagenesis effect and screening effect. Heating mutagenesis effect is occurred through the substitution of G-C base pair induced by heat, and heating screening effect produces higher forward mutation rate induced by other mutagens.
3.Initial study of acoustic radiation force impulse(ARFI) elastography quantification for evaluation of solid liver tumors
Gang WU ; Qi WANG ; Long YANG ; Jianjun YUAN ; Quan SHEN ; Fei XUE ; Bing ZHAO ; Qiang TANG ; Qiang WU
Chinese Journal of Ultrasonography 2011;20(3):226-229
ObjectiveTo evaluate the potential value of acoustic radiation force impulse (ARFI)elastography in the characterization of solid liver tumors.MethodsForty-three patients with 56 liver tumors were evaluated with ARFI,which included 21 patients with hepatocellular carcinoma (HCC),8 patients with metastase,5 patients with cholangiocarcinoma(CCC),and 9 patients with hemangioma.The shear wave velocity of the tumor and background liver parenchyma were calculated,and results were compared with 30 healthy subjects.Statistical analysis was performed on the shear wave velocity for differentiation of normal liver,background liver parenchyma,and tumors.ResultsHCC and CCC had greater stiffness than metastase (P <0.05),there were no statistical differences between HCC and CCC (P = 0.179).Malignant liver tumors had significantly greater stiffness than hemangioma and normal liver (P = 0.000).34.5% (9/26) HCC and 33.3% (4/12) hemangioma appeared softer than the background liver.With a cut-off value of 1.5 m/s for the shear wave velocity,the sensitivity,specificity,positive predictive value and negative predictive value for malignancies were 79.5%,83.3%,94.5% and 52.6%,respectively.ConclusionsARFI elastography quantification is a promising noninvasive technique for assessing solid liver tumors.Use of ARFI elastography quantification may lead to new quantitative tissue characterization parameters for differentiating hemangioma and malignant liver tumors.
4.Polypyrimidine Tract Binding Protein Negatively Regulates the Expression of HBV Surface Antigen by Interacting with HBV Postranscriptional Regulatory Element
Li-Ying CHENG ; Xiao-Hua ZHANG ; Yi LI ; Xue-Fei CAI ; Yuan HU ; Ai-Long HUANG ; Hua TANG ;
China Biotechnology 2006;0(02):-
In order to demonstrate PTB bind to HPRE,reverse transcription,PCR-mediated detection,were used.HepG2.2.15 cell line and HBs-HPRE transient expression cells were adopted to identify PTB function in HBV life cycle.The results showed that PTB could directly bind to HPRE RNA.Functional analysis indicated that PTB could inhibit the expression of HBs antigen and this inhibition was in a dose-dependent manner in HepG2.2.15 cells.Higher expression of HBs in cells transfected pcDNA3-HBs-HPRE comparing with pcDNA3-HBs,and this high expression could also be inhibited by PTB.The data demonstrated that PTB inhibits HBs expression by interacting with HPRE.
6.Screening of proteins interacting with hepatitis C virus NS3 from T7-phage display library.
Ying HUANG ; Xue-fei CAI ; Mao-rui HE ; Jun ZHANG ; Ai-long HUANG
Chinese Journal of Hepatology 2006;14(8):561-564
OBJECTIVETo screen and identify proteins that interact with hepatitis C virus NS3 by means of T7-phage display system.
METHODSHepatitis C virus NS3 was expressed by prokaryotic expression and used as a selected molecule to biopan the T7 select human liver cDNA library; the selected positive clones were identified using DNA sequencing and analyzed with BLAST program in GenBank.
RESULTSAfter BLAST analysis in all the positive clones, the proteins which interacted with the hepatitis C virus NS3 were found to be serpin peptidase inhibitor, clade A, member 1 (SERPINA1) and cyclophilin-LC.
CONCLUSIONT7-phage display system is a convenient, rapid and effective method for screening interacting proteins. The proteins thus selected will provide an important means for studying the pathogenesis and carcinogenesis of HCV.
Cell Line ; Gene Library ; Hepacivirus ; metabolism ; Humans ; Peptide Library ; Protein Interaction Mapping ; methods ; Viral Fusion Proteins ; genetics ; isolation & purification ; metabolism ; Viral Nonstructural Proteins ; genetics ; metabolism
7.Nasal reconstruction with the modified expanded forehead skin flap.
Xiao-jing LI ; Ji-long NING ; Xue-hong GAO ; Lin ZHANG ; Fei ZHU
Chinese Journal of Plastic Surgery 2005;21(3):208-210
OBJECTIVETo investigate the effect of the modified expanded forehead skin flap in nasal reconstruction.
METHODSAccording to flap design, the frontal arteries that were not selected as the pedicle were ligated in order to enhance expansion and delay effects. Besides the middle forehead skin flap for nasal reconstruction, the expanded transversal forehead flap was employed with its donor site sutured directly, resulting in inconspicuous scar. This method was used for 11 cases of nasal reconstruction.
RESULTSAll the flaps survived. Postoperative follow up for 6 months to 8 years and 4 months showed satisfactory results with good appearance and function.
CONCLUSIONSThe method of modulating the blood supply to the flap and selecting the upper area of the forehead for the flap is an effective modification for nasal reconstruction.
Adolescent ; Adult ; Female ; Forehead ; surgery ; Humans ; Male ; Rhinoplasty ; methods ; Skin Transplantation ; methods ; Surgical Flaps ; Tissue Expansion ; Young Adult
8.Establishment of a stable cell line replicating hepatitis B virus DNA carrying the reverse transcriptase region derived from a clinical isolate.
Ming-que XIANG ; Xue-fei CAI ; Wen-lu ZHANG ; Ai-long HUANG ; Jie-li HU
Acta Academiae Medicinae Sinicae 2013;35(1):13-18
OBJECTIVETo establish a stable cell line that can replicate hepatitis B virus (HBV) DNA carrying the reverse transcriptase sequence derived from a clinical isolate.
METHODSNested PCR was used to amplify the HBV DNA fragment from the serum. The fragment was cloned into a plasmid that can support HBV replication in vitro by fragment substitution reaction (FSR), followed by the cloning of the neomycin expressing fragment downstream from HBV DNA. G418 selection was conducted after the transfection of HepG2 cells with the recombinant DNA. Real-time PCR and enzyme linked immunosorbent assay (ELISA) were used to screen stable cell lines that can replicate HBV DNA, and the replication of HBV DNA by the cell line was confirmed by using Southern blot analysis.
RESULTSFragment nt55-1654 amplified from the serum DNA was substituted to the plasmid pLL, generating the plasmid p11. The neomycin fragment was cloned into p11, leading to the plasmid p11-neo, and p11-neo was confirmed to be HBV-replication-competent. A stable cell line named 3-10 that can replicate HBV DNA was obtained.
CONCLUSIONSA stable cell line was established that can replicate HBV DNA carrying the reverse transcriptase sequence derived from a clinical isolate. Real-time PCR plus ELISA may help to rapidly screen out stable cell lines replicating HBV DNA.
Cell Line ; Cloning, Molecular ; DNA Replication ; DNA, Viral ; biosynthesis ; Genetic Vectors ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Hepatocytes ; cytology ; virology ; Humans ; Plasmids ; RNA-Directed DNA Polymerase ; genetics ; Virus Replication ; genetics
9.Oridonin inhibits proliferation of Jurkat cells via the down-regulation of Brg1.
Zhen-Zhen YE ; Fei-Long XUE ; Wen-Ping DING ; Xiang KONG ; Yi-Na SHEN
Chinese Journal of Contemporary Pediatrics 2017;19(11):1208-1212
OBJECTIVETo investigate the effect of oridonin on the human acute lymphocytic leukemia cell line Jurkat and its mechanism.
METHODSJurkat cells were cultured in vitro and treated with various concentrations (0, 1.25, 2.5, 5, and 10 μmol/L) of oridonin for different lengths of time (24, 48, and 72 hours). The proliferation of Jurkat cells was analyzed by MTT assay. The changes in nuclear morphology were evaluated by fluorescence microscopy at 12 hours after treatment with various concentrations of oridonin. The expression levels of Brg1, P53, and C-myc were determined by semi-quantitative Western blot in Jurkat cells treated with various concentrations of oridonin for 24 hours or 5 μmol/L oridonin for various lengths of time (0, 2, 6, 12, and 24 hours). The expression levels of P53 and C-myc and proliferation of Jurkat cells were evaluated after Brg1 expression was knocked down by Brg1-specific siRNA.
RESULTSCompared with the control group, the proliferation of oridonin-treated Jurkat cells was significantly inhibited in a concentration- and time-dependent manner (P<0.05). According to the florescence microscopic analysis, oridonin treatment led to nuclear pyknosis in Jurkat cells. Compared with the control group, Jurkat cells treated with 5 μmol/L oridonin had reduced expression of Brg1 and C-myc but elevated expression of P53. Brg1 knock-down led to a significant reduction in proliferation of Jurkat cells (P<0.05), up-regulated expression of P53, and down-regulated expression of C-myc.
CONCLUSIONSOridonin can inhibit the proliferation of Jurkat cells, probably via the Brg1 signaling pathway.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Proliferation ; drug effects ; DNA Helicases ; analysis ; physiology ; Diterpenes, Kaurane ; pharmacology ; Dose-Response Relationship, Drug ; Down-Regulation ; Humans ; Jurkat Cells ; Nuclear Proteins ; analysis ; physiology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Proto-Oncogene Proteins c-myc ; analysis ; Signal Transduction ; physiology ; Transcription Factors ; analysis ; physiology ; Tumor Suppressor Protein p53 ; analysis
10.Performance of computer-assisted imaging system in detection of squamous intraepithelial lesion of uterine cervix.
Ying-zhuo GAO ; Yun-long HUO ; Han-xue SUN ; Ya-fei QI ; Jin-ou WANG ; Qing-jie LÜ
Chinese Journal of Pathology 2013;42(8):543-546
OBJECTIVETo evaluate the performance of computer-assisted imaging system in the detection of cervical squamous intraepithelial lesion and quality-assurance.
METHODSManual PAP screening (n = 140 580) and image-assisted screening (n = 32 885) were compared for the detection rates of squamous cell abnormalities, the atypical squamous cells (ASC) to squamous intraepithelial lesion (SIL) ratio, the positive rates of high risk human papillomavirus (HR-HPV) test in the case of atypical squamous cells of undetermined significance (ASC-US), and the correlation between cytopathology and histopathology.
RESULTSCompared with manual screening, computer-assisted imaging system showed increased overall positive detection by 0.32%, decreased detection of ASC by 0.21%, increased detection of low-grade squamous intraepithelial lesion (LSIL) by 0.22%, increased detection of high-grade squamous intraepithelial lesion or worse (HSIL) by 0.31%, and decreased ASC to SIL ratio from 2.59 to 1.60. Computer-assisted imaging system did not change the HR-HPV positive rate of the patients who were ASC-US, or the coincidence rate between cytopathology and histopathology. Moreover, the productivity of the laboratory operation increased 58.33%.
CONCLUSIONComputer-assisted imaging system significantly increases the overall positive detection rate of cervical SIL, improves accuracy and work efficiency of screening, decreases the ASC/SIL rate, and strengths the quality-assurance of laboratory testing.
Carcinoma, Squamous Cell ; pathology ; Cervical Intraepithelial Neoplasia ; pathology ; Female ; Humans ; Image Interpretation, Computer-Assisted ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; pathology ; Uterine Cervical Dysplasia ; pathology ; Uterine Cervical Neoplasms ; pathology ; Vaginal Smears ; methods