2.The application of in vivo multi-channel recording methods in the studies of the affective pain in rats.
Zhen-hua LI ; Yang YANG ; Miao-miao HOU ; Yuan WANG ; Xia QIN ; Che ZHANG ; Yu ZHANG
Chinese Journal of Applied Physiology 2016;32(1):41-45
OBJECTIVETo record the electrical activities of Antirior cingulate cortex (ACC) neurons by in vivo multi-channel recording methods using the model of complete freund's adjuvant (CFA) induced conditioned place avoidance (C-CPA), which has been set up in our previous studies.
METHODSThe electrode was self-made and the CPA responses were recorded by in vivo multi-channel recording method.
RESULTS(1) The electrical activities of ACC neurons could be successfully recorded by the self-made electrode. (2) Before or after the injection of CFA, rats were respectively conditioned to the different place. The firing rates of ACC neurons in the CFA-paired place vs that in the non-CFA-paired place was (0.853 ± 1.377) imp/s vs (0.221 ± 0.971) imp/s (P < 0.05, n = 26). (3) The CPA responses in the CFA-paired place vs that in the non-CFA-paired place were (303.55 ± 61.77)s vs (140.32 ± 33.52)s(P < 0.05, n = 6).
CONCLUSIONThe firing rates of rACC (rostral Anterior Cingulate Cortex) neurons were involved in the occurrence of the affective pain.
Animals ; Electrodes ; Freund's Adjuvant ; Gyrus Cinguli ; cytology ; Neurons ; cytology ; Pain ; diagnosis ; Pain Measurement ; methods ; Rats ; Rats, Sprague-Dawley
3.Observation of the change in ocular anterior segment after pterygium surgery with ultrasound biomicroscopy
Feng, HUA ; Zheng-Yi, LI ; Ji-Meng, CHEN ; Guo-Ting, HOU ; Li-Shan, YANG ; Yuan-Zhen, HUANG
International Eye Science 2015;(7):1280-1281
AlM: To observe the effect of anterior chamber depth and angle change after pterygium excision.METHODS:Thirty cases (30 eyes) of primary pterygium were underwent pterygium excision. Central anterior chamber depth, four direction angle open distance ( AOD ) and open angle ( AA ) were measured preoperatively and postoperatively by ultrasound biomicroscopy ( UBM ) and the intraocular pressure was observed.RESULTS:Preoperative and Postoperative intraocular pressure were 15. 17±10. 6 and 16. 23±2. 61mmHg, and the central anterior chamber depth were 2. 28±0. 39 and 2. 33± 0. 24mm. The four directions of AOD and AA were no statistical difference.CONCLUSlON:The anterior chamber depth and the angle change is not obvious after pterygium excision.
4.Advances in high-throughput transcriptome research of traditional Chinese medicines.
Zhao-Bao ZHANG ; Lin HOU ; Qing PAN ; Xu-Min WANG ; Qing-Hua CUI ; Jing-Zhen TIAN ; Lu-Yu MA
China Journal of Chinese Materia Medica 2014;39(9):1553-1558
Traditional Chinese medicine is a treasure of Chinese culture, absorbing the wisdom of the Chinese people. Continuous application of new technologies makes traditional Chinese medicine research advance with the times. After several years of development, high-throughput transcriptome study has become a mature research tool in biology. This paper reviewed the advances in medicine transcriptome study, and compared two sequencing platforms, Roche's GS FLX platform and Illumina's HiSeq 2000 platform. Moreover, this paper introduced medicine transcriptome analysis process, with Panax quinquefolius and Lonicera japonica for examples, showing the characteristics of traditional Chinese medicine transcriptome studies. High-throughput transcriptome studies facilitate traditional Chinese medicine research with overall understand of functional genes, give clear elucidation of metabolic pathways, lay molecular foundation for the traditional Chinese medicine research and offer modern interpretation for traditional Chinese medicine theory. However, the current study faces several difficulties, including weak molecular basis, high sequencing cost and staff shortages in data anaysis. In the future, with the development in sequencing technology, the combination of transcriptome and other genomics, such as proteome and metabolome, will lay a solid foundation for the new high-throughput screening and developing model for the traditional Chinese medicine industry.
Biomedical Research
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methods
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trends
;
Forecasting
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Gene Expression Profiling
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methods
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Gene Expression Regulation, Plant
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Humans
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Lonicera
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genetics
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Medicine, Chinese Traditional
;
methods
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trends
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Panax
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genetics
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Phytotherapy
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methods
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trends
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Transcriptome
;
genetics
5.Analysis of prevention of mother-to-child transmission of HIV (PMTCT) work in Zhumadian city, 2001 - 2009.
Fu-kun WANG ; Yong NIE ; Jian LIU ; Zhen-hua HOU ; Xiu-zhen JIAO ; Zhao-wen LI
Chinese Journal of Preventive Medicine 2009;43(11):988-990
OBJECTIVETo analyze the current status of maternal HIV infection, mother to child transmission, and the work accomplishments in preventing mother-to-child transmission of HIV (PMTCT).
METHODSDuring October, 2001 to May, 2009, HIV voluntary consultation and examination were carried out in 339 866 pregnant women in the urban areas, while 594 pregnant women who tested positive were intervened, and interventions were also conducted among 326 babies who were born to HIV positive mothers, including HIV immune body examination on the babies when they were 12 months and 18 months old.
RESULTSA total of 594 pregnant women were found HIV positive, with the positive rate of 0.17% (594/339 866). And the rate was declining year by year. The highest rate was 0.47% (37/7837) in 2002, and the lowest rate was 0.12% (86/73 343) in 2008. Of the 594 positive pregnant women, 228 (38.38%) terminated pregnancy voluntarily, 43 (7.24%) kept on pregnancy and 317 (53.37%) parturients. Of 326 babies born by the 317 parturients, 317 survived.298 received curbing intervention for mother to child transmission (PMTCT), the ratio was 94.01% (298/317). Of 224 babies who were 18 months old, 221 accepted examination, and 7 HIV positive. The maternal infant transmission rate after intervention was 3.17% (7/221).
CONCLUSIONThrough the prevention of mother-to-child transmission of HIV, the HIV infection status in the pregnant women can be timely observed, which can effectively decrease the level of mother-to-child transmission of HIV.
Acquired Immunodeficiency Syndrome ; prevention & control ; transmission ; Adult ; China ; Female ; Humans ; Infant ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; prevention & control ; Pregnancy
6.Three-dimensional reconstruction and anatomic variation of the portal vein based on 64-slice spiral CT data.
Wen-qiang TAO ; Zhi-dian HOU ; Da-chuan XU ; Zhen-yu BI ; Wen-hua HUANG
Journal of Southern Medical University 2011;31(1):121-124
OBJECTIVETo investigate the three-dimensional reconstruction methods of the portal vein using 64-slice spiral CT data and the anatomical variation of the portal vein.
METHODSThree-dimensional reconstruction of the portal vein was performed using Mimics software based on the 64-slice spiral CT data of 64 cases. Each model of the portal vein and its branches was evaluated according to the presentation rate, depiction quality and anatomic variation.
RESULTSThe reconstructed model showed a depiction rates of 100% for the 4-grade branches of the portal vein. The stem of the portal vein and the left and right branches of the level III or above were all displayed, but in 2 cases the superior mesenteric vein and in 1 case the spleen vein was displayed only to the level IV. Of the 64 cases, 50 (78.1%) had normal portal vein and 14 (21.9%) showed anatomical variations.
CONCLUSIONThe 3D model vividly mimics the anatomic variations of the portal vein to provide valuable information for surgical plans.
Adult ; Female ; Humans ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Portal Vein ; anatomy & histology ; diagnostic imaging ; Tomography, Spiral Computed ; methods ; Young Adult
7.Effect of TNF-alpha gene polymorphism on outcome of thalidomide-based regimens for multiple myeloma.
Juan DU ; Zhen-Gang YUAN ; Chun-Yang ZHANG ; Wei-Jun FU ; Hua JIANG ; Bao-An CHEN ; Jian HOU
Chinese Journal of Hematology 2009;30(10):649-653
OBJECTIVETo evaluate the effect of polymorphism at the -238 and -308 position of the TNF-alpha promotor region on the clinical outcome of thalidomide (Thal)-based regimens for the treatment of multiple myeloma (MM).
METHODSThe polymorphism at the -238 and -308 position of the TNF-alpha promotor region of 168 MM patients treated with Thal-based regimens were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Genotypes were tested for association with overall response by logistic regression, and survival was evaluated by univariate and multivariate analysis.
RESULTSIn TNF-alpha -238 position, 11 (6.5%) patients had GA genotype and 1 (0.6%) AA genotype. In TNF-alpha -308 position, 19 (11.3%) had GA genotype and 1 (0.6%) AA genotype. In univariate analysis, the TNF-alpha -238 GA + AA genotypes were associated with a significantly prolonged progression free survival (PFS) (P = 0.017), and a better overall survival (OS) (P = 0.150). Multivariate COX regression analysis showed that TNF-alpha -238 polymorphic status was an independent prognostic factor for prolonged PFS (P = 0.049).
CONCLUSIONThe TNF-alpha -238 polymorphic status is associated with a favorable clinical outcome in MM patients treated with thalidomide-based regimen. The polymorphism status of TNF-alpha gene might be of promise for developing a more informative stratification system for MM.
Adult ; Aged ; Aged, 80 and over ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; genetics ; Polymorphism, Genetic ; Prognosis ; Promoter Regions, Genetic ; Thalidomide ; therapeutic use ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; genetics
8.LiCl suppresses tumor growth and inhibits DNA replication in prostate cancer.
Chuan-ling HOU ; Zhen-hua ZHANG ; Di-lai HUANG ; Ai-jing SUN
Chinese Journal of Pathology 2012;41(7):475-478
OBJECTIVETo assess the effects of LiCl on prostate cancer growth and to explore the underlying mechanisms.
METHODSEffects of LiCl on cell growth in vitro and in vivo were determined by cell counting and xenografts of prostate cancer cells. Alterations in cell proliferation and the expression of DNA replication-related protein were determined by MTT assay, BrdU incorporation and Western blot.
RESULTSCompared to PBS control group, the number of prostate cancer cells (PC-3) were lower treated with 10 mmol/L LiCl, the number was 1.9×10(5), 4.8×10(5) and the difference was significant (P < 0.05). The inhibition rate of cellular proliferation were 50%, 95% and 98%, respectively, in LiCl group, NaCl and KCl control group, the difference was significant (P < 0.05). The A-Value of BrdU incorporation was 1.5, 1.3 treated with 10 mmol/L, 30 mmol/L LiCl, while the A-value of BrdU incorporation was 4 in PBS control group, the difference was significant (P < 0.05). On the protein level, LiCl downregulates expression of cdc 6, cyclins A and cyclins E, and cdc 25C, and upregulates expression of the CDK inhibitor p21(CIP1). The mean volume and weight of xenograft tumor were 50 mm(3) and 296 mg after LiCl intraperitoneal injection, But PBS control group were 180 mm(3) and 957 mg, the difference was significant (P < 0.05).
CONCLUSIONLiCl disrupts DNA replication and suppresses tumor growth of prostate cancer cells in vitro and in vivo.
Animals ; Antineoplastic Agents ; pharmacology ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin A ; metabolism ; Cyclin E ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; DNA Replication ; drug effects ; Humans ; Lithium Chloride ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Nuclear Proteins ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology ; Tumor Burden ; drug effects ; cdc25 Phosphatases ; metabolism
9.Effect of microRNA on proliferation caused by mutant HBx in human hepatocytes.
Xiao-yu FU ; De-ming TAN ; Zhou-hua HOU ; Zhi-liang HU ; Guo-zhen LIU ; Yi OUYANG ; Fei LIU
Chinese Journal of Hepatology 2012;20(8):598-604
To study the effect of micro (mi)RNA on cellular proliferation induced by hepatitis B x protein, HBx, in human liver cells and to investigate the underlying molecular mechanism of this cancer-related effect. The human L02 hepatocyte cell line was stably transfected with HBx (L02/HBx) or an HBx mutant (L02/HBx-d382) that induces higher levels of cellular proliferation. The differential miRNA expression profiles were determined by microarray analysis and confirmed by real-time PCR. Two miRNAs, miR-338-3p and miR-551b, that were found to be significantly down-regulated in the L02/HBx-d382 cells were selected for further study and transfected individually into cells using the lipofectamine procedure. The cell survival rate was analyzed by MTT assay, and cell cycles were assessed by flow cytometry. Expressions of cyclinD1, cyclinG1, and E2F1 were assessed by real-time PCR and Western blotting. Compared with the microarray miRNA profile of L02/pcDNA3.0 cells, six miRNAs were up-regulated and five miRNAs were down-regulated in the L02/HBx-d382 cells, while four miRNAs were up-regulated and 12 were down-regulated in the L02/HBx cells. The microarray results were consistent with real-time PCR results. Transfection of miR-338-3p and miR-551b significantly inhibited the cell survival rates (P less than 0.001) and induced G0/G1 phase cycle arrest. According to MTT results: for L02/HBx-d382 cells, compared with lipofectamine or non-transfected (NC) controls, the t value of miR-338-3p was 10.402, 9.133 and the t value of miR-551b was 8.763, 7.403; for L02/HBx cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 9.105, 8.074 and the t value of miR-551b was 7.673, 7.52. According to flow cytometry results: for L02/HBx-d382 cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 12.173, 11.107 and the t value of miR-551b was 15.364, 13.377; for L02/HBx cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 15.416, 13.378, and the t value of miR-551b was 13.276, 13.109. The protein levels of cyclinD1, cyclinG1, and E2F1 were significantly reduced by both miR-338-3p and miR-551b ( P less than 0.001). For L02/HBx-d382 cells, compared with lipofectamine or NC controls: E2F1 had t = 11.132, 10.031 and 12.017, 10.973, respectively; cyclinD1 had t = 15.654, 15.013 and 15.447, 14.733, respectively; cyclinG1 had t = 8.017, 7.661 and 7.402, 7.417, respectively. For L02/HBx cells, compared with lipofectamine or NC controls: E2F1 had t = 14.244, 13.331 and 15.022, 14.468, respectively; cyclinD1 had t = 8.695, 8.137 and 7.877, 7.503, respectively; cyclinG1 had t = 7.73, 7.471 and 7.596, 7.41, respectively. In contrast, the mRNA levels for E2F1, cyclinD1, and cylcinG1 showed no significant differences between the miRNA transfected cells and controls. Wild-type HBx and the high proliferation-inducing mutant HBx can influence the miRNA expression profile of L02 cells. HBx down-regulates miR-338-3p and miR-551b in L02 cells, and the high proliferation-inducing mutant has a more robust effect. The mechanism of miR-338-3p- or miR-551b-mediated cell growth inhibition appears to be related to the direct modulation of cyclinD1, cyclinG1, and E2F1.
Blotting, Western
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Carcinoma, Hepatocellular
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genetics
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metabolism
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pathology
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Cell Cycle
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Cell Line
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Cell Proliferation
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Cyclins
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genetics
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metabolism
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Gene Expression Regulation, Neoplastic
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Genes, Viral
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Hepatitis B virus
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genetics
;
metabolism
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Hepatocytes
;
metabolism
;
pathology
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Humans
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Liver Neoplasms
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genetics
;
metabolism
;
pathology
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MicroRNAs
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genetics
;
metabolism
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Mutation
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Oligonucleotide Array Sequence Analysis
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RNA, Messenger
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genetics
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Real-Time Polymerase Chain Reaction
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Trans-Activators
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genetics
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metabolism
;
Transfection
10.Expression of the human papillomavirus type 16L/E7 fusion protein in E. coli and observation of its immunogenicity in mice.
Hou-wen TIAN ; Zhen-mei YE ; Zhen-hua LU ; Ren JAO ; Tao BIAN ; Li ZHAO ; Li RUAN
Chinese Journal of Experimental and Clinical Virology 2006;20(2):33-37
BACKGROUNDMany epidemiological and experimental evidences prove that cervical cancers are strongly associated with genital high-risk types of human papillomavirus (HPV). HPV16 is present in 50% of the tumor specimens. Thus, it is important to develop vaccines against HPV16 and cervical cancer. The authors studied the expression of the HPV16 L1DeltaCE7N fusion protein in E. coli and observed its immunogenicity.
METHODSThe fragment of HPV16 L1DeltaC gene and the E7N gene were amplified by PCR separately; the fusion gene named L1DeltaCE7N was generated by fusing E7N to the C terminal of L1DeltaC then the chimeric gene was cloned into prokaryotic expression vector pGEX-2T and expressed in E. coli strain JM109. The L1DeltaCE7N protein expressed were detected by Western blot. Finally its immunogenicity was characterized in immunized mice.
RESULTSIt was proved that the sequence and open reading frame of fusion gene L1DeltaE7N was correct by sequencing; SDA-PAGE gel analysis showed that HPV16 L1/E7 fusion protein was highly expressed in E. coli; the protein was expressed as soluble form and the molecular weight was about 85 x 10(3). The fusion protein could be purified by affinity chromatography and gel filtration. The ELISA result indicated that L1/E7 could elicit specific antibodies against L1 and E7 in immunized mice. In vivo tumor protection test indicated that tumor formation was retarded or prevented in the mice after vaccination with L1/E7, when C57 BL/6 mice were challenged by syngeneic HVP16E6 and E7 transformed tumor cells.
CONCLUSIONHPV16L1/E7 fusion protein was expressed in E. coli, it can be a candidate for prophylactic and therapeutic vaccine for HPV16-associated infection and tumors.
Animals ; Blotting, Western ; Cell Line, Tumor ; Escherichia coli ; genetics ; Female ; Humans ; Immunization ; methods ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Neoplasms, Experimental ; immunology ; pathology ; prevention & control ; Oncogene Proteins, Fusion ; genetics ; immunology ; metabolism ; Oncogene Proteins, Viral ; genetics ; immunology ; metabolism ; Papillomaviridae ; genetics ; immunology ; metabolism ; Papillomavirus Infections ; immunology ; pathology ; prevention & control ; Papillomavirus Vaccines ; administration & dosage ; immunology ; Recombinant Fusion Proteins ; immunology ; metabolism ; ultrastructure