2.Effects of tetramethylpyrazine on fibrosis of atrial tissue and atrial fibrillation in a canine model of congestive heart failure induced by ventricular tachypacing.
Yazhou LIN ; Chunxuan XU ; Yulian DENG ; Lin CHEN ; Hai HUANG ; Jian DU
Journal of Integrative Medicine 2006;4(1):35-8
To explore the effects of tetramethylpyrazine (TMP) on fibrosis of atrial tissue and atrial fibrillation in a canine model of congestive heart failure (CHF) induced by ventricular tachypacing.
3.Effects of Different Compatibilities of Ramulus Cinnamomi and Peony in Guizhi Decoction on Diabetic Cardiac Autonomic Neuropathy.
Xiao LI ; Jin-long YANG ; Du-fang MA ; Hai-qing LIN ; Xiang-dong XU ; Hua Jiang YUE
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(6):741-745
OBJECTIVETo observe the preventive effect different compatibilities of Ramulus Cinnamomi (RC) and peony in Guizhi Decoction (GD) on diabetic cardiac autonomic neuropathy (DCAN).
METHODSTotally 60 male rats were randomly divided into 5 groups, i.e., the blank control DM group, the model group, the methycobal group, the 1:1 (RC/peony) Guishao group, the 2:1 Guishao group, and the 1:2 Guishao group, 10 in each group. Rats were pretreated with corresponding drugs for 1 week, and then induced diabetes by intraperitoneal injection of STZ. Drugs were administrated by gastrogavage for 4 more weeks after STZ-injection. Enzyme-linked immunosorbent assay (ELISA) was employed to detect levels of tyrosine hydroxylase (TH), choline acetyltransferase (CHAT), nerve growth factor. (NGF), and ciliary neurotrophic factor (CNTF) in myocardial homogenates.
RESULTSAfter 4-week modeling, body weight (BW) was obviously lower, but blood glucose (BG) was higher in STZ rats than in rats of the blank control DM group. There was no statistical difference in BW or BG among the 5 groups (P >0.05). Compared with the blank control group, TH, TH/CHAT, and NGF in left ventricle and ventricular septum increased, CHAT and CNTF increased in the model group (P < 0.05, P < 0.01). Compared with the model group, TH and TH/CHAT in left ventricle decreased (P < 0.05, P < 0.01), CNTF in left ventricle increased (P < 0.05), CHAT in left ventricle and ventricular septum increased (P < 0.05, P < 0.01) in the methycobal group. TH and TH/CHAT in left ventricle and ventricular septum decreased, CNTF in left ventricle and ventricular septum increased (P < 0.05, P < 0.01), CHAT in left ventricle and ventricular septum increased (P < 0.01), NGF in ventricular septum decreased (P < 0.01) in the 1:1 Guishao group. TH/CHAT in left ventricle decreased (P < 0.01), CHAT and CNTF in left ventricle and ventricular septum increased (P < 0.05, P < 0.01) in the 1:2 Guishao group. Compared with the methycobal group, CHAT in left ventricle decreased, TH and TH/CHAT in left ventricle increased in the 2:1 Guishao group (P < 0.05, P < 0.01). TH and TH/CHAT in ventricular septum decreased (P < 0.05), CHAT and CNTF in left ventricle and ventricular septum increased (P < 0.05, P < 0.01) in the 1:1 Guishao group. Compared with the 1:2 Guishao group and the 2:1 Guishao group, CHAT in left ventricle increased, TH/CHAT in left ventricle decreased, TH and TH/CHAT in ventricular septum decreased, CHAT in ventricular septum increased, CNTF in left ventricle and ventricular septum also increased in the 1:1 Guishao group (all P < 0.01).
CONCLUSIONSSTZ model rats had autonomic neural injury, manifested as lowered vagal nerve activity and hyperactive sympathetic nerves. GD could effectively suppress hyperactive cardiac sympathetic nerves and protect the vagus. Besides, GD (1:1) showed the optimal effect in regulating the balance of cardiac autonomic nerves and could be used in early prevention of DCAN.
Animals ; Blood Glucose ; Choline O-Acetyltransferase ; Diabetic Neuropathies ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Heart ; Heart Ventricles ; Male ; Myocardium ; Nerve Growth Factor ; Paeonia ; Rats ; Tyrosine 3-Monooxygenase
4.Pilot study of intravesical instillation of two new generation anthracycline antibiotics in prevention of superficial bladder cancer recurrence.
Si-Yang CHEN ; Lin-Dong DU ; Yu-Hai ZHANG
Chinese Medical Journal 2010;123(23):3422-3426
BACKGROUNDSuperficial bladder cancer accounts for 60% - 70% of all bladder cancer cases in China, when treatment consists of only transurethral resection of the bladder tumor (TUR-BT), recurrence and progresses in the bladder are observed in some patients. There are numerous reports of trials of intravesical instillation of anticancer agents with the objective of lowering this recurrence rate. The aim of this study was to compare the prophylactic efficacy and safety of epirubicin (EPI), pirarubicin (THP) and hydroxycamptothecin (HCPT) in superficial bladder cancer.
METHODSThis study enrolled a total of 189 patients who had been diagnosed with superficial bladder cancer during the period from 2004 through 2007 at Beijing Friendship Hospital. All patients were randomly allocated to one of three treatment groups. Patients in group A received 29 doses of EPI 30 mg/30 ml, patients in group B received 29 doses of THP 30 mg/30 ml, and patients in group C received 29 doses of HCPT 30 mg/30 ml, over a period of 24 months.
RESULTSThe recurrence-free rate in the 2 anthracycline treatment groups (A and B) were significantly better than that of the HCPT treatment group. In the safety evaluation, the incidences of pollakiuria, pain on urination, dysuria, hematuria, and contracted bladder were not significantly different between groups A and B, but some were significantly higher in groups A and B than that in group C.
CONCLUSIONThe efficacy of EPI and THP was significantly better than HCPT in the prevention of bladder cancer recurrence.
Administration, Intravesical ; Adult ; Aged ; Anthracyclines ; administration & dosage ; adverse effects ; Antibiotics, Antineoplastic ; administration & dosage ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; prevention & control ; Pilot Projects ; Urinary Bladder Neoplasms ; prevention & control
5.Construction of the subtracted cDNA library of striatal neurons treated with long-term morphine.
Bo BAI ; Hai-qing LIU ; Jing CHEN ; Ya-lin LI ; Hui DU ; Hai LU ; Peng-li YU
Chinese Medical Sciences Journal 2011;26(1):54-59
OBJECTIVETo construct a morphine tolerance model in primarily cultured striatal neurons, and screen the differentially expressed genes in this model using suppression subtractive hybridization (SSH).
METHODSSbtracted cDNA libraries were constructed using SSH from normal primarily cultured striatal neurons and long-term morphine treated striatal neurons (10-5 mol/L for 72 hours). To check reliability of the cell culture model, RT-PCR was performed to detect the cAMP-responsive element-binding protein (CREB) mRNA expression. The subtracted clones were prescreened by PCR. The clones containing inserted fragments from forward libraries were sequenced and submitted to GenBank for homology analysis. And the expression levels of genes of interest were confirmed by RT-PCR. Results CREB mRNA expression showed a significant increase in morphine treated striatal neurons (62.85 ± 1.98) compared with normal striatal neurons (28.43 ± 1.46, P < 0.01). Thirty-six clones containing inserted fragments were randomly chosen for sequence analysis. And the 36 clones showed homology with 19 known genes and 2 novel genes. The expression of 2 novel genes, mitochondrial carrier homolog 1 (Mtch1; 96.81 ± 2.04 vs. 44.20 ± 1.31, P < 0.01) and thymoma viral proto-oncogene 1 (Akt1; 122.10 ± 2.17 vs. 50.11 ± 2.01, P < 0.01), showed a significant increase in morphine-treated striatal neurons compared with normal striatal neurons.
CONCLUSIONSA reliable differential cDNA library of striatal neurons treated with long-term morphine is constructed. Mtch1 and Akt1 might be the candidate genes for the development of morphine tolerance.
Analgesics, Opioid ; pharmacology ; Animals ; Cells, Cultured ; Corpus Striatum ; cytology ; Drug Tolerance ; physiology ; Gene Expression Profiling ; Gene Library ; Molecular Sequence Data ; Morphine ; pharmacology ; Neurons ; cytology ; drug effects ; Nucleic Acid Hybridization ; methods ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; methods
6.Transfection of human hepatic stimulator substance gene could protect BEL-7402 cells against hepatotoxins.
Li CHEN ; Hong-liu SUN ; Lin YANG ; Hai-jun DU ; Wei AN
Chinese Journal of Hepatology 2004;12(2):99-101
OBJECTIVETo investigate protective effects of hHSS transfection against CCl4 or H2O2.
METHODScDNA coding for hHSS was constructed into eukaryotic vector of pcDNA3.1 and transfected into BEL-7402 hepatoma cells. The expression of hHSS was analyzed with Northern blot.
RESULTSThe growth of the hepatoma cells was remarkably enhanced 24 to 144h after hHSS gene transfection, which suggesting hHSS gene expression could stimulate cells activity. Meantime, incubation of both wild-type and vector-transfected as well as hHSS-transfected cells with CCl4 or H2O2 resulted in severe damage as marked by cell mortality and the rate of apoptosis. However, it appeared that the transfection of hHSS enabled the hepatoma cells to raise obvious resistance against CCl4 and H2O2 injury. Compared the vector cells to the vector-transfected cells, apoptosis ratio were (32.44+/-0.52)% and (25.60+/-0.66)% in which treated with CCl4, while (47.78+/-0.45)% and (37.40+/-0.69)% in which treated with H2O2, t value is 16.82 and 25.20, P<0.01. MAPK phosphorylation was also activated after HSS transfected.
CONCLUSIONThe function of hHSS gene expression could be related to proliferation of cell and protection against free radical damage.
Apoptosis ; drug effects ; Carbon Tetrachloride ; toxicity ; Cytoprotection ; Free Radicals ; Growth Substances ; genetics ; physiology ; Humans ; Hydrogen Peroxide ; toxicity ; Liver Neoplasms ; pathology ; Mitogen-Activated Protein Kinases ; metabolism ; Peptides ; genetics ; physiology ; Phosphorylation ; RNA, Messenger ; analysis ; Transfection
7.Studies on anti-endotoxin activity of F022 from Radix Isatidis.
Ai-hua LIN ; Shu-xian FANG ; Jian-guo FANG ; Guang DU ; Yun-hai LIU
China Journal of Chinese Materia Medica 2002;27(6):439-442
OBJECTIVETo study the anti-endotoxin activity and mechanism of F022 from Radix Isatidis.
METHODThe production of TNF-alpha and IL-6 of murine peritoneal macrophages stimulated by LPS was measured by ELISA. The temperature in rabbits was tested after i.v. administration of LPS. The lethality of BCG-primed mice was induced by LPS.
RESULTIf F022 was added to macrophages culture simultaneously with LPS or 1 h before addition of LPS, production of TNF-alpha and IL-6 by macrophages was remarkably inhibited in vitro. F022 inhibited the fever induced by LPS in rabbits and protected BCG-primed mice from LPS induced lethality if given before administration of LPS.
CONCLUSIONThe anti-endotoxin effect of F022 may inhibit LPS binding to its receptor, and it may be a LPS receptor antagonist.
Animals ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Fever ; chemically induced ; drug therapy ; Interleukin-6 ; secretion ; Isatis ; chemistry ; Lipopolysaccharides ; antagonists & inhibitors ; Macrophages, Peritoneal ; secretion ; Male ; Mice ; Mice, Inbred C57BL ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rabbits ; Tumor Necrosis Factor-alpha ; secretion
8.Polymorphism of human killer cell immunoglobulin-like receptor gene in Chinese Jiangsu Han population.
Yu-Dong DAI ; Rui-Ping DONG ; Hai-Lin DU
Journal of Experimental Hematology 2010;18(3):776-779
This study was purposed to investigate the distribution of killer cell immunoglobulin-like receptor(KIR)in Jiangsu Han population of China. KIR genomic typing were analyzed by PCR-SSP typing methods in 269 randomly unrelated healthy individuals. The results showed that all 16 known KIR genes were found in Jiangsu Han population, the total KIR genes found in Jiangsu Han population were 34 genes. Out of 34 KIR found genotypes, 15 genotypes (AA1, BX2, BX3, BX4, BX5, BX6, BX8, BX9, BX11, BX13, BX33, BX68, BX69, BX70, BX75) had also been identified in Zhejiang and Hong Kong populations, otherwise, 19 genotypes (BX10, BX12, BX17, BX23, BX26, BX27, BX28, BX31, BX35, BX42, BX47, BX57, BX72, BX74, BX79, BX154, BX188, BX231, BX370) had never been observed in Zhejiang and Hong Kong populations. The rare allele BX42 (detected only in Greece population) and BX231 (detected only in Sweden population) and BX370 (detected only in Macedonia population) were identified in Jiangsu Han population. In conclusion, all 16 known KIR genes were detected in Jiangsu Han population, the total 34 KIR genotypes are found in Jiangsu province, among them the BX42, BX231 and BX370 are rare KIR genotypes.
Asian Continental Ancestry Group
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genetics
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China
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Gene Frequency
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Haplotypes
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Humans
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Polymorphism, Genetic
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Receptors, KIR
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genetics
9.Experimental study of the inhibiting effect of the lentiviral vector mediated herpes simplex virus-thymidine kinase/ganciclovir on GVHD.
Kai-lin XU ; Feng ZHU ; Bing DU ; Fei GAO ; Hai CHENG ; Xiu-ying PAN
Chinese Journal of Hematology 2007;28(5):303-307
OBJECTIVETo study the effect of lentiviral vector mediated herpes simplex virus-thymidine kinase/ganciclovir (HSV-TK/GCV) on graft- versus-host disease (GVHD) after allogeneic bone marrow transplantation (allo- BMT) in mice.
METHODSDonor splenic lymphocytes from C57BL/6 which were infected by lentiviral vectors carrying HSV-TK were transplanted into 60Co gamma ray irradiated recipient mice with donor bone marrow cells. GCV 25 mg x kg(-1) x d(-1) was administered in 3 groups on day 0, +7, +12 respectively after transplant for 7 days by intraperitoneal injection. Survival time, severity of GVHD, incidence of GVHD, T lymphocytes immune reconstruction and of allogeneic chimerism ratio were detected after allo-BMT.
RESULTSThe average survival times for GCV 0 day, +7 day and +12 day group were (30. 10 +/- 5.21) d, (36.40 +/- 5.28) d and (28.20 +/- 4.82) d respectively, being significantly longer than that in the control group [(15.10 +/- 0.43) d] (P < 0.05). The 50 d-survival rate for TK/GCV + 7 day group was 60%. While for 0 day and +12 day group was 40% and 30% respectively. The incidence of grade III approximately IV GVHD in the control group was 100%, and the dead mice in experimental groups showed pathological changes of II approximately III GVHD. Long-term alive recipient mice only developed grade I approximately II GVHD after allo-BMT. The number of CD4+ lymphocytes in experimental groups was higher than that in control group (P <0.05), but CD8+ lymphocytes was lower on day +5, +10, +15 day (P <0.05). Allogeneic chimerism rate of recipient mice on +30 d was 100%.
CONCLUSIONSHSV-TK/GCV induced by the lentiviral vectors has a definite effect in prevention of GVHD after allo-BMT. GCV administrated from 7 days post-transplantation showed the best effects.
Animals ; Bone Marrow Transplantation ; immunology ; Ganciclovir ; pharmacology ; Genetic Vectors ; Graft vs Host Disease ; prevention & control ; Lentivirus ; genetics ; Lymphocyte Transfusion ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Simplexvirus ; enzymology ; Thymidine Kinase ; genetics ; Transfection ; Transplantation, Homologous
10.Research of the p65 gene function in the prostate cancer cell by the obtaining of shRNA sequences blocking the expression of nuclear factor kappa- B (p65) stably and construction of lentivirus vector
Hai HUANG ; Tao DU ; Jian HUANG ; Tianxin LIN ; Caixia ZHANG ; Wen DONG ; Xinbao YIN ; Zhenghui GUO ; Kewei XU ; Chun JIANG ; Jinli HAN
Chinese Journal of Urology 2010;31(6):386-390
Objective To obtain shRNA sequences that can stably block the expression of Nuclear Factor kappa- B (p65) in the prostate cancer cell line LNCaP and construct the lentivirus vector.And validate the gene function of p65 in the cell line. Methods According to p65 genetic information, we design siRNA1, siRNA2, siRNA3 those three siRNA sequences targeting the ods area of p65 gene and then form the corresponding four pairs of complementary single strand DNA of shRNA, including the sense strand and the antisense strand. The synthetic shRNA sequence was inserted into the empty pSIH1-H1-copGFP shRNA Vector, and after transfecting the prostate cancer cells , the inhibitory effect of p65 mRNA by different sequences was detected through real-time PCR, and the inhibitory effect of p65 protein expression was detected by Western-blotting. Thus we can obtain highly effective shRNA sequences in the inhibition of p65 in prostate cancer cells. MTT, flow cytometry, transwell were chosen to test the cell growth, migration and invasive power in vitro to compare the difference of the experimental group, control group and negative group. Results The third shRNA sequence had the best inhibitory effect and the inhibitory effect of p65 mRNA in prostate cancer cell line was 59 % and the protein was 81%. It's position locates in p65 (NM_021975 ) 1096-1113 and it's stemloop sequence is 5'-GATCCGCCCTATCCCTTTACGTCATTCAAGAGATGACGTAAAGGGATAGGGCTTTTTG-3'. After transfecting, the prostate cancer cell line had the low expression of p65 stably. Through MTT, we got the growth curve, which showed that the growth ability of experimental group was significantly decreased compared with the control group and the Logarithmic growth didn't appear in the first 96 hours. Flow cytometry test displayed that the percentage of G0-G1-phase cells in experimental group was 61.49%, and the control group was 44.89%, idle group was 41.52%, which was increasing oberviously. The S-phase cells in the experimental group was 28.58%, compared with the 47.36% and 46. 10% diminished. The results of transwell showed that the experimental group had 16. 5000±6. 62076 cells and the other two groups had 45. 6333 13. 54159 and 36. 8333±5. 68412 cells, which showed the invasive power of experimental group was significantly declined(P<0.05).Conclusions It's successful to obtain shRNA sequences that can stably block the expression of p65 in the prostate cancer cell line LNCaP and construct the lentivirus vector. p65 can positively regulates the biological behavior of prostate cancer LNCaP cell line in the cell growth, migration and invasive power.