1.CT perfusion imaging study of perihematomal cerebral blood flow in experimental intracerebral hemorrhage
Jian ZHOU ; Pei-yi GAO ; Xiao-guang LI ; Hong WAN
Chinese Journal of Rehabilitation Theory and Practice 2004;10(8):472-474
ObjectiveTo study the mechanism of hemodynamic alternation surrounding the hematoma in the acute stage of intracerebral hemorrhage (ICH) in rats.MethodsSeventy male Sprague Dawley rats were randomly divided into ICH group and sham operated group, which were microinjected with 40 μl fresh autologous blood or saline into the right caudatum respectively. The each group was divided into 7 subgroups at 1h,3h,6h,12h,24h,48h and 72h after the ICH. CT perfusion imaging in measurements of regional cerebral blood flow adjacent to hematomas was performed. The ratios of side to side were measured at the regions around the hematomas by personal computer aided mapping. So the parameters of regional cerebral blood flow(rCBF), regional cerebral blood volume(rCBV) and mean transit time(MTT) were calculated respectively.ResultsThe rCBF and rCBV adjacent to the hematomas were lower than those of the outer region pronouncedly. The alternation of rCBF around the hematoma were fluctuated, which reduced to the valley at 1h after ICH, and then gradually returned to the peaks at 6h and 24h after ICH. In the meantime, the rCBV around the hematoma reduced to the valley at 1h after ICH, and then gradually increased to the peak at 24h after ICH.ConclusionThe abnormal hemodynamic changes can be found in the perihematomal region after ICH. The alternation of rCBF around the hematomas are fluctuated, but the changes of rCBV remain continuous increase. The mass effect of hematoma, intracranial hypertension caused by the mass effect of hematoma, and the autoregulation of cerebral blood flow motivated by the initial depression of cerebral blood flow play a very important role in the changes of cerebral blood flow.
5.Establishment and application of neutralization assay based on H5N1 avian influenza pseudotyped virus in vitro
Guoliang ZHANG ; Boping ZHOU ; Cheguo CAI ; Xinchun CHEN ; Guilin YANG ; Jian LU ; Guang NIE ; Baoluo ZHOU
Chinese Journal of Clinical Infectious Diseases 2011;04(2):91-95
Objective To establish neutralization assay based on H5N1 avian influenza pseudotyped virus in vitro and to evaluate neutralizing titer of convalescent serum from 2 patients with H5N1 avian influenza.Methods pHR-Luc,pCMV△8.2 and CMV/R-SH or CMV/R-TH were cotransfected into 293T cell by co-precipitation with calcium phosphate.Pseudotyped virus supernatant was harvested 72 h posttranofection and identified the expression of HA and P24 by Western blot,and then we analyzed infective activity of 200 μL supernatant of pseudotyped virus.293T cell integrated HA was prepared and anti-HA antibodies in convalescent serum were measured with FACS assay.Neutralizing titers of convalescent serums against Shenzhen and Thailand pseudotyped virus were determined based on calculating IC50 with neutralizing assay.Results Pseudotyped virus involved P24 and HA,and precursor protein HA0 could cleavage into HA1 and HA2 with biological activity.Pseudotyped virus possessed better infective activity,and RLA value was about 2 × 104 with 200 μL supernatant.Both convalescent serums contained anti-HA antibodies and had cross-reactivity against different virus clades with FACS assay.Both convalescent serums had neutralizingactivity and could cross-neutralize different virus clades.However,both serums'neutralizing titers against Shenzhen virus were higher than Thailand.Conclusion We successfully constructed infectious pseudotyped virus which integrated HA of Shenzhen or Thailand virus,and it could be used for evaluation of serum neutralizing activity fast,efficiently and safely with broadly application prospect.
6.Construction of pseudotype retrovirus which integrates hemagglutinin of H5N1 avian influenza virus isolated from human in Shenzhen
Guoliang ZHANG ; Boping ZHOU ; Xinchun CHEN ; Cheguo CAI ; Jian LU ; Guilin YANG ; Guang NIE ; Baoluo ZHOU
Chinese Journal of Microbiology and Immunology 2009;29(1):53-57
Objective To construct pseudotype retrovirus which integrates hemagglutinin(HA)of H5N1 avian influenza virus(AIV)isolated from human in Shenzhen.Methods AIV HA gene was amplified bv RT-PCR,then it was ligated with pGEM-T vector,and identified by restriction enzyme digestion and sequenced.HA gene was cloned into CMV/R vector at the site of Sal Ⅰ and BamH Ⅰ.pHR-Luc,pCMV&8.2 and CMV-HA were co-transfected into 293T cell by co-precipitation with calcium phosphate.The pseudotype virus supernatant was harvested 72 h post-transfection and ultracentrifugation,and the HA and P24 expression on the surface of pseudotype virus was analyzed by western blot.Meanwhile.the infection activity of HIV-HA pseudotype virus was identified in different kinds of cell lines,including MDCK,HeLa,CHO and 293T.Results A/Shenzhen/406H/06 belonged to subclade2.3 with open reading frame(ORF)of HA gene encoded 567 amino acides,whose accession number was EF137706 in GenBank.HA gene was cloned into CMV/R successfully.After co-transfection of above vectors,it revealed that HA protein could integrate pseudotype virus by western blot,and precursor protein HA0 could cleavage into HA1 and HA2 with biological activity.Finally.HIV-HA pseudotype virus could infect 4 kinds of cell lines,which indicated its property of infectivity and catholicity.Conclusion The pseudotype retrnvirns wassuccessfully constructed,which can integrate HA protein of A/Shenzhen/406H/06 and had property of infectivity.It call be used in the further research,including selection of neutralizing antibodies and epitope analysis.
7.Optimal treatment for malignant glaucoma
Zhi-Jian, HUANG ; Wen-Qiang, ZHANG ; He-Zheng, ZHOU ; Guang-Jie, HAN
International Eye Science 2015;(1):141-143
Abstract?AlM:To investigate the choice of different treatments for malignant glaucoma.? METHODS: ln this retrospective case series, 21 malignant glaucoma patients ( 21 eyes ) admitted in Wuhan General Hospital of Guangzhou Military Command from May 2012 to May 2013 were analyzed. Sixteen eyes ( 76%) developed malignant glaucoma after filtration surgery, 3 eyes ( 14%) after EX - PRESS glaucoma filtration device, 2 eyes ( 10%) after glaucoma filtration Ahmed valve implantation. Main Outcome of corrected visual acuity, intraocular pressure ( lOP ) , anterior chamber depth and complications were detected.?RESULTS: lOP recovered by drug control in 13 eyes, anterior chamber depth. Four eyes were treated by vitreous water- bag aspiration combined with anterior chambers reconstructing. Two eyes were treated by cataract extraction and intraocular lens implantation. Two eyes were treated by posterior capsule excision combined with anterior vitrectomy. lOP before and after treatment was 29. 81±4. 98, 12. 71±3. 77mmHg, respectively (P=0. 00). Anterior chamber depth before and after treatment was 0.41± 0. 34, 2. 13 ± 0. 54mm, respectively (P = 0. 00). Corrected visual acuity before treatment was 0. 19 ± 0. 17, after treatment was 0. 20±0. 16 (P= 0. 36). Except for vitreous hemorrhage in 1 eye, there were no ocular or systemic adverse events observed in all patients.? CONCLUSlON: lt is good to diagnose malignant glaucoma in early period, and treated it step by step. For this can reduce lOP and restore anterior chamber.
8.Relationship between Spondyloppiphyseal Dysplasia Tarda Gene Escaping X Chromosome Inactivation and Spondyloppiphyseal Dysplasia Tarda Phenotype
chao, GAO ; huai-li, WANG ; qiang, LUO ; guang-yao, SHENG ; jian-hua, ZHOU ; tie-zheng, GAO
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To explore the relationship between X - linked spondyloepiphyseal dysplasia tarda (SEDL) gene escaping X chromosome inactivation( XCI) and SEDL phenotype. Methods RT - PCR was performed on total RNA which was isolated from blood samples of patients, female carriers and controls. Patients and female carriers were selected from the pedigree with SEDL caused by the mutation (IVS2 - 2A→C) of the gene. cDNA was analyzed by polyacrylamide gelelectrophoresis(PAGE). Results PAGE data indicateed that female carriers expressed both normal and mutant SEDL mRNA,meaning the SEDL gene escaping XCI. Family investigation showed carrier females in the SEDL pedigree presented no symptoms. Conclusions The SEDL gene escaping X chromosome in-activation is firstly identified from human body. This may explain that carrier females present no symptoms.
9.Oral administration of insulin inhibits islet beta cell apoptosis and prevents diabetes in NOD mice.
Tie-Jian JIANG ; Zhi-Guang ZHOU
Journal of Central South University(Medical Sciences) 2007;32(4):615-619
OBJECTIVE:
To investigate the effect of oral administration of insulin on insulitis beta cell apoptosis and diabetes in non-obese diabetic (NOD) mice, and to explore the mechanism of immune tolerance induced by insulin.
METHODS:
Eighty-six female NOD mice were randomly divided into an insulin group (n=43) and a phosphate buffered saline (PBS) group (n=43). From 4 weeks of age, the recombinant human insulin (Humulin R) 1 mg (70 microL) was administrated in the oral insulin group and 70 microL PBS in the control group respectively, twice per week before 12 weeks of age and then once weekly until 30 weeks. Insulitis and beta cell apoptosis of islets were observed at 12 weeks. IL-4 and IFN-gamma in the sera were measured by enzyme linked immunosorbent assay (ELISA). The expression levels of I-Abeta(g7), IL-4, IFN-gamma, IL-1beta, Fas and TGF-beta mRNA of islets, and IL-4, IFN-gamma, TGF-beta mRNA of Peyer's patch were measured by reverse transcription-polymerase chain reaction (RT-PCR) at 12 weeks.
RESULTS:
The incidences in the insulin group were significantly lower than those in the PBS group (55.6% vs 85.7% at 30 weeks, 70.4% vs 96.4% at 52 weeks, P<0.05). The insulitis scores in the insulin group were lower than those in the PBS group, but there was no statistical significance. Fas expression on islets and apoptotic beta cell rates in the insulin group were lower than those in the PBS group (P<0.05). In the insulin group, serum IL-4 levels were higher, and IFN-gamma levels were lower than those in the PBS group (P<0.05). The levels of I-Abeta(g7), IFN-gamma, IL-1beta and Fas mRNA transcription in islets and IFN-gamma mRNA transcription in Peyer's patch were both lower in the insulin group, and IL-4, TGF-beta mRNA levels were higher than those in the PBS group (P<0.05).
CONCLUSION
The specific autoantigen insulin may induce the immune tolerance and prevent the diabetes in NOD mice, but it cannot block the progression of insulitis. Oral administration of insulin can induce the regulatory T cells, and make Th1 to Th2 cytokine shifts in the system and islets, thus preventing the Fas-mediated beta-cell apoptosis and diabetes.
Administration, Oral
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Animals
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Apoptosis
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drug effects
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Cytokines
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metabolism
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Diabetes Mellitus, Type 1
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drug therapy
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pathology
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Female
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Insulin, Regular, Human
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administration & dosage
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pharmacology
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Islets of Langerhans
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cytology
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drug effects
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Mice
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Mice, Inbred NOD
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Th1-Th2 Balance
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fas Receptor
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metabolism
10.Study of chemical constituents in stem rind of Daphne giraldii.
Guang-Xiong ZHOU ; Yong-Chun YANG ; Jian-Gong SHI
China Journal of Chinese Materia Medica 2006;31(7):555-557
OBJECTIVETo study the constituents with the pain-relieving activity from the stem rind of Daphne giraldii.
METHODThe partition of the ethanol extract and chromatographic separation of the fractions were carried out by the monitoring of anelgesic pharmacological activity. The structures of the isolated compounds were determined by MS and NMR.
RESULTFour compounds were isolated from the pain-relieving fraction. Three of them were identified as diterpenes, gniditrin (1), gnidicin (2) and daphnetoxin (3). Compound 4 was determined as Z-octadecyl caffeate.
CONCLUSIONCompounds 1, 2 and 4 were isolated from the plant for the first time.
Analgesics ; chemistry ; isolation & purification ; pharmacology ; Caffeic Acids ; chemistry ; isolation & purification ; pharmacology ; Daphne ; chemistry ; Diterpenes ; chemistry ; isolation & purification ; pharmacology ; Heterocyclic Compounds, 4 or More Rings ; chemistry ; isolation & purification ; pharmacology ; Molecular Structure ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry