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PCD-hIL-2 and Chinese medicine accelerate the immunofunction reconstitution in bone marrow transplantation mice

Journal of Peking University(Health Sciences).2001;33(2):178-180，182.

Objective: In order to study how to accelerate the reconstitution immunofunction in BMT mice, first of all, we established a immunodeficiency model of BMT in BALB/C mice. Then BMT mice were injected with PCD-hIL-2 directly into skeletal muscle, and treated with traditional Chinese medicine. Methods: The experiment groups are designed as（A）Chinese medicine + PCD-hIL-2;（B）PCD-hIL-2;（C）Chinese medicine +hIL-2;（D）Chinese medicine;（E）hIL-2;（F）BMT;（G）normal control;（H）radiation control. Results: We compared groups A B C D to E or F groups, found（1）The splenocytes/thymocytes count increase obviously.（2）Killing activity of NK cell rises obviously in vivo.（3）The response of splenocytes、thymocytes、BM cells to mitogen goes up.（4）The reactivity of splenocytes to foreign IL-2 goes up. （5）CFU-GM count is increased. Conclusion: The expression of hIL-2 is very low by nude DNA injection ,but it is enough to have biological function and therapeutic effect .If only Chinese medicine was applied, the immunological condition was obviously recovered.

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The expression and role of IL-6, IL-6R and acute phase reaction factor in peripheral blood mononuclear cells of multiple myeloma

Xiaojun HUANG ; Xuan DUAN ; Yan LI ; Hong LIN ; Janfan FU ; Daopei LU

Journal of Peking University(Health Sciences).2001;33(2):170-172.

Objective： To study the expression and role of IL-6, IL-6 receptor(IL-6R) and acute phase reaction factor (APRF) in PMNC of multiple myeloma(MM) patients. Methods:The bioactivity of IL-6 and the level of sIL-6R in sera of MM patients were measured using IL-6 dependent cell line 7TD1 and ELISA respectively. The APRF activity of PMNC DNA binding protein was observed by gel blocking electrophoresis (EMSA). Results:The bioactivity of IL-6 and the level of sIL-6R in MM sera were significantly higher than those of the control and the level was coordinated with that of the tumor. The results of PMNC DNA binding protein EMSA showed that PMNC of MM patients expressed APRF after stimulation of IL-6,whereas there was no activity of APRF in the normal PMNC. Conclusion:In MM,some regulation machines of IL-6 signal transduction pathways changed (eg.protein kinase), resulting in the activation of transcription of acute phase reaction element(APRE), APRF may have a role in pathogenesis of MM.

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The expression and purificiation of human Annexin Ⅴ overexpressed in E. coli and the detection of apoptosis

Chunhui DI ; Qun SHI ; Yingmei ZHANG ; Cheng ZHAO ; Yingcheng ZHONG ; Dawu HUANG ; Dalong MA

Journal of Peking University(Health Sciences).2001;33(2):167-169.

Objective: in order to provide rapid and reliable method. Methods: Encoded Annexin Ⅴ cDNA was amplifyed from U937 cDNA libary by PCR and then subcloned into E coli expression vector. MS2-Annexin Ⅴ fusion protein could be overexpressed in E　coli. The MS2 bacteria protein could be removed by thrombin digestion.The mature Annexin Ⅴ was obtained by ion exchange chromatography and the FITC labled Annexin Ⅴ could be used in the detection of apoptosis. Results:Up to 37% of the total bacterial proteins was rhAnnexin Ⅴ as showed by SDS-PAGE. The purification of Annexin Ⅴ is over 99%. The FITC labled Annexin Ⅴ could efficiently detect apoptosis. Conclusion: We successfully established the technique procedure of obtaining a large quantity of Annexin Ⅴ and provided the basic routine for popularizing the detection of apoptosis' with high effciency.

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The mechanism of protective effects of melatonin on global ischemia/reperfusion induced brain injury in rats

Yu RAO ; Baoshan KU

Journal of Peking University(Health Sciences).2001;33(2):164-166.

Objective: To study the effects of melatonin（MT） on inducible nitric oxide synthase (iNOS) expression in hippocampal CA1 at 24 hours and TDT-mediated dUPT nick end labling (TUNEL) positive cells in hippocampal CA1 at 48 hours after global ischemia（20 min）/ reperfusion in rats. Methods: MT was injected intraperitoneally at 0 h, 1 h, 2 h and 6 h after ischemia（20 min） induced by the“occlusion of four arteries”, 2.5 mg*kg－1 or 10 mg*kg－1 each time, respectively. Researching the expression of iNOS in hippocampal CA1 with the immunocytochemistry method 24 hours after reperfusion and the number of TUNEL positive cells in hippocampal CA1 48 hours after reperfusion utilizing TUNEL. Results: At these doses, MT could decrease iNOS expression in hippocampal CA1 at 24　h after global ischemia （20 min）/reperfusion and the number of TUNEL cells in hippocampal CA1 at 48h after reperfusion in rats. Conclusion: Decreasing the expression of deleterious iNOS maybe one of the mechanisms involved in protective action of MT on ischemia-vulnerable brain region.

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Multi-groups controled study on the association of Helicobacter pylori infection with gastric cancer and stomach diseases

Yongwen JIANG ; Runtian WANG ; Tao WANG ; Jieping ZHANG ; Daonian LEI

Journal of Peking University(Health Sciences).2001;33(2):160-163.

Objective: This multi-group case control study was performed to determine whether Helicobacter pylori infection was associated with gastric cancer and stomach diseases. Methods: Gastric cancer and stomach disease patients that were diagnosed pathologically between 1994 and 1996 years were divided into several case groups and control group, all histological specimens collected in gastric cancer high risk region of Shandong Province were used to measure exposure history. Results: After variables concerning age, sex and education were adjusted, odds ratios of chronic superficial gastritis in antrum, peptic ulcer disease and active degree of inflammation associated with H. pylori infection remained significant (OR was 2.072, 2.980, 2.086 respectively). Correlation analyses showed negative relationship between degree of stomach diseases and H. pylori infection(r=－0.217). Conclusion: The results support the hypothesis of a relationship between H. pylori infection and the development of chronic superficial gastritis in antrum, peptic ulcer disease and active degree of inflammation.

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Characterization of the antagonistic effect on α1-adrenoceptor of the Chinese herb medicine Xiao Long Tong Bi

Zhizhen LU ; Yinzeng LI ; Qide HAN ; Jinming JIA

Journal of Peking University(Health Sciences).2001;33(2):157-159.

Objective: To better understand the antagonistic effect of Xiao Long Tong Bi (XLTB), a Chinese herb medicine, on α1-adrenoceptor (α1-AR). Methods: (1) Radio ligand binding assay . Specific　125I-BE2254(2-β(4-hdroxyphenyl)-ethyl amino-methyl-tetralone) binding was measured by incubating membrane of canine cerebral cortex with a single concentration of　125I-BE2254 in the presence of 15 concentrations of XLTB. Half-effectual concentration of inhibition (IC50) and Hill coefficients (nH) were determined by Hill plots. (2) Contractile responses of rat prostate strip in vitro were determined. pKB values for XLTB in competitively inhibiting NE-stimulated contraction of tissues were measured by the method of Ainlakshana. Results: XLTB competitively inhibited binding of　125I-BE2254 to α1-AR in a concentration -dependent manner. IC50 values for XLTB in canine cerebral cortex were (34.0±6.0)　g*L－1, the Hill efficiency value (0.7±0.1) was significantly decreased from unity. Contractile studies showed that XLTB competitively antagonized the NE concentration-response curve with pKB values of (37.0±11.0) g*L－1 or (30.0±8.0) g*L－1 when XLTB concentration was 70 g*L－1 or 170 g*L－1, respectively. The pKB values for XLTB in antagonizing NE-induced contraction of tissues were showed to fit in well with the IC50 values on rat prostate. Conclusion: These results suggest that XLTB appears to be a competitive antagonist for α1-AR.

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The effect of an inhibitor of inducible nitric oxide synthase on the embryo development of mice

Junmei WANG ; Mingqing SONG ; Bin LIU

Journal of Peking University(Health Sciences).2001;33(2):153-156.

Objective: To determine whether aminoguaidine（AG）,a selective inhibitor of inducible nitric oxide synthase (NOS), affect the embroy development of mice, and to study the mechanisms of iNOS affecting on pregnancy. Methods: AG(10 mg*kg－1*d－1 or 20 mg*kg－1*d－1) were injected subcutaneously to mice from day 3，7，14. kill mice at day19. The weight of fetus and placenta, the number of fetus and fetal resorptions were measured. Histological changes of placenta and umbilical cord were observed. Immunohistochemistry and NADPH histochemistry were adopted to study NOS activity of placental and umbilical cord respectively. Results: The conceptus in the uterus were resorbed in the early pregnancy. In the early and middle pregnancy the weight of pregnant mouse and the number of fetus of AG groups was decreased. The number of fetal resorptions was increased. But the weight of fetus and placenta were not affected. The expression of iNOS in the placenta were no difference significantly between the two groups with image analysis. Conclusion: AG inhibits the implantation and growth of embryo in the early and middle pregnancy, buy doesn't affect the formed fetus and placenta significantly.

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Oxidized low density lipoprotein stimulated endothelial nitric oxide by activating lectin-like oxidized-LDL receptor

Yaqin XU ; Junhua ZHANG ; yang KE ; jing CAO ; Tao NING ; Liya FENG ; Chaoshu TANG

Journal of Peking University(Health Sciences).2001;33(2):150-152.

Objective:To explore the role of lectin-like oxidized low density lipoprotein (ox-LDL) receptor(LOX1) in ox-LDL stimulating endothelial nitric oxide(NO) production. Methods: LOX1 mRNA expression was detected by reverse transcription-polymerase chain reaction(RT-PCR). The quantity of NO was detected by Enzyme-method. Results: Incubation of ox-LDL increased endothelial NO production and LOX1 mRNA expression. When HUVECs were incubated with ox-LDL as well as the inhibitor of LOX1, polyinosinic acid,the increase of NO production and LOX1mRNA expression were attenuated. Conclusion: Ox-LDL enhanced endothelial NO production in a concentration-dependent manner, and the effect of ox-LDL on endothelial NO was mediated by LOX1.

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Effect of urotensin Ⅱ on nitric oxide synthese/nitric oxide pathway in rat aorta

Zhisheng JIANG ; Yonggang ZHANG ; Yongfen QI ; Song XU ; Shaoping XU ; Yongzheng PANG ; Chaoshu TANG

Journal of Peking University(Health Sciences).2001;33(2):147-149.

Objective: To study the effect of urotensin(U Ⅱ) on nitric oxide synthase(NOS)/nitric oxide(NO) system in rat aorta. Methods: The aortic slices were incubated in vitro. The medium nitrite content, vascular NOS activities and cGMP content were measured. Results: After incubation of aorta for 1.5－3.0 h, UⅡ(10－9－10－7　mol*L－1) increased nitrite production ( 14.8%－80.9%, P<0.01) in dose-dependent manner. U Ⅱ significantly stimulated the total NOS activities, mainly activated constitutive NOS (P<0.05 or P<0.01), however, the inducible NOS activities were not altered (P>0.05). Incubation of aortic slices with U Ⅱ (10－9－10－7 mol*L－1) increased vascular cGMP content in dose-dependent manner. Co-incubation vascular tissue with U Ⅱ and　GN-L-nitro-arginine, an inhibitor of NOS, obviously inhibited the U Ⅱ-induced NOS activation, NO production and cGMP formation. Conclusion: The results suggest that U Ⅱ can activate the vascular NOS/NO pathway, increasing tissue NO production and cGMP content.

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Helicobacter pylori induces the synthesis of cyclooxygenase-2 in human gastric epithelial carcinoma cell line BGC-823

Yongchang CHEN ; Jianzhong ZHANG ; Tie LI ; Tao SHENG ; Wenyu ZHU

Journal of Peking University(Health Sciences).2001;33(2):144-146.

Objective: To investigate the effect of Helicobacter pylori (H. pylori)　on the synthesis of cyclooxygenase-1 (COX-1)　and cyclooxygenase-2 (COX-2) in gastric epithelial cells. Methods: a VacA(+) and CagA(+) international standard H. pylori line NCTC11637 and a human gastric epithelial carcinoma cell line BGC-823 were used. Western Blotting was applied to detect the synthesis of cyclooxygenase. Results: The content of COX-2 protein increased obviously after the cells were incubated with H. pylori sonicating extract for 1　h and the increase lasted for at least 6　h whereas the content of COX-1 protein did not change during the incubation with H. pylori extract. H. pylori lipopolysaccharide (LPS) had no effect on COX-2 synthesis. Conclusion: H. pylori stimulated the synthesis of COX-2 in BGC-823 cells and the effect was LPS-independent.

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