1.The effects of attenuated Salmonella carrying IL-2/NK4 double gene on immune function in mice
Chinese Journal of Biologicals 2024;37(5):571-576
Objective To study the effects of attenuated Salmonella(Ty21a-pIRES-IL-2-NK4,TPIN)carrying interleukin-2(IL-2)/4-kringle antagonist of hepatocyte growth factor(NK4)double gene on humoral and cellular immune function.Methods Eighteen BALB/c mice,half male and half female,were randomly divided into control group(1. 5 mL 10%NaHCO3 by gastric tube feeding),Ty21a group(0. 1 mL Ty21a by gastric tube feeding)and TPIN group(0. 1 mL TPIN by gastric tube feeding),with 6 mice in each group. The immunization was boosted twice 7 d after the initial immunization. At 21d after administration,the blood samples were collected from eyeballs and the serum was separated,which was detected for the serum IgG antibody level by ELISA. The thymus and spleen of mice were isolated aseptically,and the spleen cells were stimulated by Ty21a and TPIN respectively in vitro. After 72 h,the proliferation ability of spleen cells was measured by CCK-8 assay,and the expression level of cytokines in spleen cells was detected by ELISA. The spleen and thymus were weighed,the spleen and thymus indexes were analyzed,and HE staining was performed.Results Compared with the control group and Ty21a group,the serum IgG level(F = 111. 74,P < 0. 01)and the contents of IFNγ,IL-4 and IL-10 in spleen cell supernatant(F = 38. 21,11. 37 and 26. 92,respectively,each P < 0. 05)increased significantly,as well as the spleen and thymus indexes(F = 10. 419 and 5. 859,respectively,each P < 0. 05)showed significant increase. In mice of Ty21a and TPIN group,the thymus cortex widened,lymphocytes increased,and there was mild inflammatory reaction;the white pulp and lymphocytes in spleen increased with neutrophil infiltration.Conclusion TPIN has a good immune protective effect,and can significantly stimulate the body to produce humoral immunity and cellular immunity,which may have a good therapeutic effect on tumors.
3.Changes of ICAM-1 expression in transplanted lung at the early stage after rat pulmonary transplantation
Chinese Journal of Organ Transplantation 2000;21(4):213-215
Objective To study the changes of ICAM-1 expression in transplanted lung at the early stage after rat pulmonary transplantation.Methods The expression level of ICAM-1 protein and its mRNA in rat transplanted lung after 4 h of ventilation and reperfussion were detected by using immunohisto chemical method(SABC method)and semiquantitative reverse transcriptase polymerase reaction method in the experimental group(n=6).When donor lungs had been flushed with low potassium solution(LPDS) and preserved in 4.C LPDS for 12 h,left orthotopic pulmonary transplantation was performed.In control group (n=6), left pulmonary artery,pulmonary vein and bronchi were fully freed to be naked.Immunohistochemical test results were recorded in negative(-),suspicion(±),faint positive(+),positive(++)and intensive positive(+++).PCR products were separated by agrose gels and the density of the bands were determined by density scanning.Results The stained color of alveolar epithelial cells and pulmonary vascular endothdial cells in the transplanted lung was significantly more intensive in experimental group than in control group(P<0.01) and the relative density values(ICAM-1/βactin)were also significandy higher in experimental group(0.837±0.044) than in control group (0.442±0.037),P<0.01. Conclusions ICAM-1 expression in the transplanted lung was up-regulated in the early stage after pulmonary transplantation,which was related with the enhancement of ICAM-1 mRNA.
4.Role of bright vessel sign on raw three dimensional arterial spin labeling image in evaluating occlusion of intracranial artery
Zhihua XU ; Benqiang YANG ; Yang DUAN
Chinese Journal of Neurology 2017;50(5):354-357
Objective To explore the role of bright vessel sign (BVS) on raw three dimensional arterial spin labeling (3D ASL) image in evaluating occlusion of intracranial artery. Methods One hundred and twenty-two patients who were highly suspected of acute cerebral infarction were enrolled and analyzed. All patients were performed magnetic resonance scan with diffusion weighted image (DWI), 3D ASL and magnetic resonance angiography (MRA) sequences within 24 hours after admission. The presence or absence of restricted lesion on DWI, BVS and occlusion of intracrainal artery on MRA was reviewed. The sensitivity, specificity, positive predictive value, negative predictive value of BVS and consistency of BVS and MRA in assessing occlusion of intracranial artery were assessed. Results The sensitivity, specificity, positive predictive value and negative predictive value of BVS in assessing occlusion of intracranial artery were 83%, 99%, 95% and 96%, respectively.And the presence or absence of BVS on ASL was highly consistent with MRA in assessing occlusion of intracranial artery (κ=0.86, P<0.01). Conclusion BVS has a good sensitivity and high specificity in identifying occlusion of intracranial artery, and it is highly consistent with MRA.
5.Problems and countermeasures for management of medical equipment of CPAPF's mobile forces
Zhihua YANG ; Yonggang LI ; Guochen YANG
Chinese Medical Equipment Journal 2004;0(07):-
The management of medical equipment is a difficult part of health works for CPAPF'smobile health institutions,and it restricts the development of health institutions in a certain extent.So in order to do it well,this paper puts forward some countermeasures for the existing new problems in the management of medical equipment of CPAPF's mobile forces,whose aim is to enhance the ability of medical support for CPAPF's mobile forces.
6.Migration of olfactory ensheathing cells and their effects on neurological function recovery in rats with cerebral infarction
Zhihua YANG ; Wenli SHENG ; Minjian WANG
Chinese Journal of Tissue Engineering Research 2008;12(34):6759-6762
BACKGROUND: Olfactory ensheathing cell (OEC) transplantation can promote the recovery of neurological function in rats with cerebral infarction, while the migratory pattern of transplanted OECs and the relationship between OECs migrated to various encephalic regions and plasticity upregulation and recovery of neurological function of the encephalic region are still unknown.OBJECTIVE: To observe the migratory pattern and therapeutic value of OEC transplantation in rats with cortical cerebral infarction.DESIGN, TIME AND SETTING: Randomized, controlled, cell transplantation observation experiment was conducted between June 2002 and January 2004 at the Laboratory of Department of Neurology, the First Affiliated Hospital of Sun Yet-San University and Animal Experimental Center of SUN YET-SEN University, Guangzhou, Guangdong Province, China.MATERIALS: Sprague-Dowley rats, 2.5-month-old, were used for olfactory ensheathing cell culture. 150 Sprague-Dowley rats,60-90 days, were used to replicate stroke-prone reuovascular hypertensive rat model using two-kidney two-clip method.METHODS: We purified OECs with differential time adherent method and labeled OECs with Hoechst 33342 before transplantation. Seventy stroke-prone renovascular hypertensive rats were used to prepare middle cerebral artery occlusion model.model rats were randomly allocated to 3 groups to receive OEC transplantation: peri-mfarct cortex transplantation group,contralateral cortex transplantation group and bilateral transplantation group.MAIN OUTCOME MEASURES: The recovery of motor and sensory function was observed at 2 weeks and 6 weeks after transplantation with behavior and sensory function examination; the survival and distribution conditions of transplanted ceils were observed under the fluorescence microscope.RESULTS: The recovery condition of motor and sensory function of rats in bilateral transplantation group was obviously better than that of rats in peri-infarct cortex transplantation group and contralateral cortex transplantation group (P < 0.01), nerve fiber number and positive signal value of growth associated protein-43 in the marginal zone of cerebral infarction were also more than that in peri-infarct cortex transplantation group and contralateral cortex transplantation group. Transplanted cells in peri-infarct cortex transplantation group migrated to infarct and contralateral cortex along corpus callosum, transplanted cells in contralateral cortex transplantation group migrated to midline and infarct cortex along corpus callosum, transplanted ceils in bilateral transplantation group migrated along corpus callosum and could be seen in bilateral cortices while more in infarct cortex.CONCLUSION: Transplanted OECs can survive for a long time period, and these cells not only confine to injection point but also can migrate to infarct and contralateral cortex along corpus caliosum to promote the recovery of neurological function of rats with cerebral infarction, the effect is more significant in bilateral transplantation.
7.Infant with both alpha 1 antitrypsin deifciency and biliary atresia:acase report and literature review
Ying YANG ; Yan LIU ; Zhihua HUANG
Journal of Clinical Pediatrics 2016;34(8):610-613
Objectives To study the clinical characteristics and early diagnosis of infant with both alpha 1 antitrypin deficiency (α1-ATD) and biliary atresia (BA). Methods The clinical characteristics, serum biochemical parameters, gene mutations and treatment of one infant with both α1-ATD and BA was reported. Related literatures about liver disease caused by α1-ATD were reviewed and analyzed. Results The infant was characterised with neonatal cholestasis, hepatomegaly, elevated serum ALT, AST, total bilirubin (TB), direct bilirubin (DB) and γ-glutamyltransferase (γ-GT) and absence of bile secretion from the duodenal drainage tube. BA was conifrmed by laparotomy and pathological examination and Kasai′s operation was performed. Further, the infant was confirmed by SERPINA 1 gene mutation analysis, which leads to the diagnosis of α1-ATD. The case of infant with both alpha 1 ATD and BA has not yet been reported at home and abroad. According to the literatures, children with α1-ATD were characterized with cholestasis, hepatomegaly, hypoproteinemia, high serum ALT and AST, coagulation disorders caused by vitamin K 1 deifciency and hepatic dysfunction. Prognosis was poor without early diagnosis and treatment. Conclusions For infant cholestasis, a lot of auxiliary examinations should be performed to identify the etiology of cholestasis. Gene analysis could help differential diagnosis. Prompt diagnosis and early treatment are the key to improve the survival rate and prognosis.
8.Effects of low-concentration lidocaine on persistent sodium currents in hypoxic isolated rat CA1 hippocampal neurons
Yi ZHANG ; Yilin YANG ; Zhihua JIAO
Chinese Journal of Anesthesiology 1996;0(08):-
Objective To investigate the effects of low-concentration lidocaine on the persistent sodium currents enhanced by hypoxia in isolated rat CA1 hippocampal neurons. Methods Brains were harvested from 10-14 day old SD rats of both sexes. Hippocampi were immediately isolated and cut into slices (400-500 ?m) which were incubated in artificial cerebral-spinal fluid (ACSF) at 31 ℃ for 1-1.5 h. CA1 regions were isolated and hippocampal neurons were prepared by enzymatical digestion. The experiment was performed in 7 groups ( n = 10 each): hypoxie control group (C) and lidocaine 1, 3, 6, 10, 20, 30 ?mol groups (L1-6). The isolated neurons were transferred to the recording chamber. The persistent sodium currents were recorded using whole-cell patch clamp technique first under normal condition. The normal perfusion solution was then replaced with hypoxie and glucose free perfusion solution within 20 seconds. The persistent sodium currents were recorded again after being perfused with hypoxie and glucose free solution with and without lidocaine. Results The persistent sodium current was greatly enhanced after 5 min hypoxia as compared to the baseline value before hypoxia. The persistent sodium current in group L1-6 was significantly lower than that in group C after 5 min hypoxia. The inhibitory effect of lidocaine on the persistent sodium current enhanced by hypoxia was dose-dependent. Conclusion Low concentration lidocaine can inhibit the persistent sodium current enhanced by hypoxia.
9.Expression and Characterization of Human/Murine Chimeric Antibodies with Specificity for CD3 Antigen
Zhihua YANG ; Decheng SHEN ; Xiangyang LIU
Chinese Journal of Cancer Biotherapy 1995;0(02):-
Exons encoding the variable regions of the light and heavy chain of the murine McAb HIT3a against CD3 antigen were isolated from HIT3a gene library and inserted into mammalian expression vectors containing the human K and y1 region exons. The chimeric genes were transfected into murine SP2/0 hybridoma cells by Lipofectin. Antibody levels of culture supernatants from transfectomas ranged 21~32 ?g/ml. The chimeric antibodies bound specifically to human T cells and competed effectivelly with the parental anti-CD3 murine McAb for binding to CD3 antigen on human T cells. The ability to promote antibody-dependent cell-mediated cytolysis was significantly enhanced with the chimeric antibodies as compared with anti-CD3 murine McAb. The mitogenic activity of human T cells can be suppressed and enhanced by the anti-CD3 chimeric antibodies. The cytotoxicity to tumor cells and proliferation of human T cells mediated by the anti-CD3 chimeric antibodies were much higher than IL-2 alone. Chimeric HIT3a antibody is a clinically relevant, genetically engineering antibody with potential use in treatment of graft-versus-host disease in tranplantation, autoimmune diseases and some tumors.
10.Construction of Anti-CD3 Human/Murine Chimeric Antibody Genes for Expression in Mammalian Cells
Zhihua YANG ; Yiwan LI ; Decheng SHEN
Chinese Journal of Cancer Biotherapy 1995;0(02):-
For construction of anti-CD3 human/murine chimeric antibody genes, a selective first-strand cDNA synthesis from mRNA or RNA of murine McAb HIT3a was performed using murine Ig constant region primers, and then cDNA of heavy and light chain variable domains of murine immunoglobulin were amplified by PCR using a set of degenerated oligonucleotide primers. Using these cDNA fragments as probes, the L and H chain V region exons encoding the murine McAb anti-CD3 were isolated from the gene library of HIT3a DNA and inserted into mammalian expression vectors containing the human ? and yl constant region exons for construction of human/murine chimeric antibody genes.