2.Clinical Analysis on 54 Children with Lennox-Gastaut Syndrome
yi-ming, CHAI ; shui-zhen, ZHOU ; dao-kai, SUN
Journal of Applied Clinical Pediatrics 2004;0(08):-
Objective To study the etiology,clinical characteristics,electroencenphalography(EEG),mental degree of Lennox-Gastaut syndrome(LGS). Method Retrospectively analyzed etiology,sex,age,seizure types,EEG,mental degree of 54 children diagnosed as LGS. Results The number of male was 36,female was 18,seizure onset from 1 month to 8 years,diagnosing age from 3 months to 11years. The EEG reveals 1.5-2.5 Hz spike-wave discharges and a slow baseline activity. Conclusions LGS is one of the most difficult epilepsys to treat and need frequently more than 2 antiepilepsy drugs. It is characterized by variable etiology,multiple types of intractable seizures, and has enormous detrimental effects on patient′s developmental health.
3.The anti-oxidant effect and the possible mechanism of plcroside Ⅱ in cerebral ischemia/reperfusion injury in rats
Li SUN ; Ling WANG ; Lei FANG ; Huanmin GAO ; Zhen LI ; Xiaodan LI ; Zhen ZHOU
International Journal of Traditional Chinese Medicine 2011;33(9):803-806
Objective To investigate the anti-oxidant effect and the possible mechanisms ofpicrodide II in cerebral ischemia/reperfusion injuries in rats. Methods A total of 90 adult, healthy, mmale Wistar rats were used to established the middle cerebral artery occlusion reperfusion (MCAO/R) models by intraluminal monofilament suture on the left external-internal carotid artery. The treatment group and the positive control group were respectively injected with 1.0% picroside II (10 mg/kg, 250 μl) and salvianic acid A sodium (10 mg/kg, 250 μl) via the tail vein, and the negative control group and sham-surgery group were injected with 0.1mol/L phosphate buffer saline (PBS) 250 μl. The neurological deficit scores were evaluated with Bederson's test. The cerebral infarction volume was observed with tetrazolium (TTC) staining. The apoptosis positive cells were counted by terminal deoxynucleotidyl transferase dUTP nick-end labeling and the expressions of inducible nitric oxide synthase (iNOS) and superoxide dismutase (SOD) were detected with immunohistochemical assay.The concentration of iNOS and SOD proteins in brain tissue was detected by enzyme linked immunosorbent assay.Results Neurological behavioral malfunction appeared in all the rats with MCAO/R. The infarction focuses emerged in the ischemic hemisphere following the MCAO/R injuries. The number of apoptotic cells and the expression of iNOS increased while the SOD reduced after MCAO/R. After the treatment of picrodide Ⅱ, the nervous behavioral function (1.28±0.38)improved, the infarction volume(68.73±4.46)% reduced, the number of apoptosis positive cells(6.10± 1.26), the expressions and the concentrations in brain tissue of iNOS(4.67+0.51)decressed while those of SOD (0.53 ±0.14) increased significantly compared with the negative control groups(t=3.16、 2.51、 4.15、3.12、 3.25, P<0.05). Conclusion PicrodideⅡ might play a neuroprotective effect by inhibiting the neuronal apoptosis and the expressions of iNOS and SOD after cerebral ischemia/reperfusion injuries.
4.Establishment and Phenotype Analysis of SHBG Knockout Mouse Model
Yiping SUN ; Yue WANG ; Zhen JIN ; Xiaoyan WANG ; Lei SUN ; Xuan ZHANG ; Chong FENG ; Xiaohua ZHOU
China Biotechnology 2017;37(8):39-45
Objective:To generate sex hormone binding globulin(SHBG) conditional knockout mice model.In order to investigate the physiological function of SHBG in vivo and to provide experimental means for the study of the relationship between SHBG and gestational diabetes mellitus.Methods:The mouse genomic DNA sequence of SHBG was verified through bioinformatic analysis.According to the SHBG genomic DNA sequence,the gene targeting and knockout vector were constructed.Transfection of the vectors to ES cells by electroporation was performed according to common protocol.Positive ES cells were screened and identified by PCR.Therefore,the dual selected ES cells were microinjected into blastula,then blastula transplantations into the host mice.The chimeric mice were mated with C57BL/6J mice,and the Flox mice were obtained after screening.The Flox mice were hybridized with EIIA-Cre transgenic mice,and the progeny of the SHBG gene knockout (SHBG-/-) mice were obtained by autocopuation for several times.Results:Several Flox homozygous mice and SHBG gene knockout mice were successfully obtained.Compared with control mice,homozygous mice of SHBG gene knockout were well developed and had reproductive ability.The growth and development of SHBG knockout mice were not significantly different from that of wild type mice.Conclusion:Homozygous mice model of SHBG gene knockout was successfully established,which laid the foundation for further study of the role of SHBG in the gestational diabetes.The SHBG gene knockout mouse model was successfully established and the preliminary phenotypic analysis was performed,which laid the foundation for further study on the role of SHBG in gestational diabetes mellitus.SHBG gene knockout mice were normal in appearance.Due to the limited number of samples and many unknown biological characteristics of gene knockout mice,it needs further study.
5.Inhibitory effects of RNA interference on expression of matrix metalloproteinase-2 gene and malignancy behaviours of ovarian cancer cells
Qingyuan SONG ; Xiujie SHENG ; Yingqun ZHOU ; Zhen LI ; Man SUN ; Zhihui WANG
Cancer Research and Clinic 2012;24(4):224-228
Objective To investigate the inhibitory effects of RNA interference (RNAi) on the expression of matrix metalloproteinase-2 (MMP-2) gene and growth, adhesion,invasiveness and migration of ovarian cancer cells. Methods One specific target sequence of MMP-2 gone and one non-specific sequence (NC group) were chosen,the medium DMEM as blank group.After transfection of ovarian cancer OVCAR-3 cells, the RT-PCR and Western blot were used to detect mRNA and protein expression of MMP-2 gene, the growth ability was detected by MTT assay, the abilities of adhesion was detected by cell adhesion assay, the invasion and migration were detected by Matrigel invasion assay and wound healing assay. Results By contrast to the NC group,the mRNA expression was decreased by 73.8 %,78.8 % and 78.4 %(P< 0.05) in 24 h,48 h and 72 h after transfection and protein expression was decreased by 72.6 %,81.2 % and 76.4 %(P< 0.05) respectively at the same time. The 48 h group had the most efficient inhibitory effect. Cell growth curve revealed that cell growth was not significantly inhibited (P> 0.05). Adhesion was significantly reduced,the inhibitory rate was 55.0 % at 60 min and 44.8 % at 90 min (P< 0.05),respectively. Invasion and migration were significantly reduced as well,the inhibitory rate on invasion and migration were 29.7 % and 35.8 %(P<0.05), respectively. Conclusion siRNA mediated MMP-2 down-regulation in ovarian OVCAR-3 cells can inhibits its adhesion,invasion and migration,but do not significantly affect its growth,suggesting a important target to ovarian cancer gene-therapies.
6.Methodological study of quantitative detection of Mycobacterium tuberculosis by immunomagnetic capture combined with PCR-ELISA
Zhen WANG ; Yuhua GONG ; Caidi QIAN ; Chunhong SUN ; Liping ZHOU ; Xingli FU ; Qixing SHAO
International Journal of Laboratory Medicine 2014;(21):2931-2933
Objective To establish a quantitative detection method for Mycobacterium tuberculosis by immunomagnetic capture combined with PCR-ELISA detection system with double internal standards(IMC-PCR-ELISA) .Methods The immunomagnetic (Dynabeads? ) which could specifically capture Mycobacterium tuberculosis were prepared .According to Mtp40 and IS6110 gene sequence of Mycobacterium Tuberculosis ,2 pairs of primers(upstream primer was modified with Biotin at 5′end) ,2 same-length mutant fragments with PCR amplified fragments ,and 3 capture probes(modified with digoxigenin at 3′end) were designed .Myco-bacterium tuberculosis were captured by immunomagnetic ,then detected by PCR-ELISA with double internal standards .Results The IMC-PCR-ELISA method could yield quantitative results in about 4 h with a detection limit at 5 × 103 copies/mL .There was a fine linear relationship between the copies of Mtp40(IS6110)in fact and in the calculation through formula when the concentrations of low internal standards were 30-70 copies/mL and the concentrations of high internal standards were 8 000-12 000 copies/mL (r2 =0 .998) .No nonspecific amplification was observed .Conclusion A rapid and quantitative method for the detection of Myco-bacterium tuberculosis was established successfully .The IMC-PCR-ELISA method was rapid ,sensitive ,secific and quantitative .
7.Migration and homing of bone marrow mesenchymal stem cells in segmental nerve injury
Xuefeng ZHOU ; Zhiwu REN ; Ming LU ; Yu WANG ; Zhen SUN ; Jiang PENG
Chinese Journal of Tissue Engineering Research 2015;(28):4465-4471
BACKGROUND:A large number of studies have confirmed that tissue-engineered stem cel therapy is feasible to repair peripheral nerve injury, but the repair mechanism is unclear.
OBJECTIVE:To observe the differentiation and homing of bone marrow mesechnymal stem cel s under local nerve microenvironment by exploring the migration and effect of bone marrow mesenchymal stem cel s in the repair of damaged nerve.
METHODS:Male SD rats, aged 8 weeks, were selected to establish segmental nerve injury models by freezing the sciatic nerve. Thirty-six model rats were randomized into three groups (n=12):frozen nerve injury group, cel injection into the nerve group, cel injection around the nerve group. Before modeling and at 4, 8, 12 weeks after cel implantation, the sciatic nerve function index was measured. Electrophysiological test, contractility recovery rate, wet weight recovery rate of the triceps surae were detected and Masson staining was performed;toluidine blue staining of the distal nerve injury and immunofluorescence staining of the damaged nerve were performed.
RESULTS AND CONCLUSION:At 4, 8, 12 weeks after cel implantation, the sciatic nerve function index was ranked as fol ows:frozen nerve injury group
8.Aβ25-35 protein in different concentrations to create a rat model resembling Alzheimer's disease and the differences in their learning and memory ability
Kai WANG ; Qiang LI ; Weiming SUN ; Jiachun XU ; Wei GUO ; Yang CAO ; Zhen ZHOU
Chinese Journal of Comparative Medicine 2017;27(3):14-19
Objective To observe the learning and memory ability of rats after injection of Aβ25-35 protein in different concentrations into the lateral ventricle assessed by Morris water maze test, and to explore the optimal concentration of Aβ25-35 in the preparation of AD model rats.Methods Male SD rats were randomly divided into sham operated group and model group.The rats of model group received Aβ25-35 injection in concentrations of 2 μg/μL, 4 μg/μL and 8 μg/μL, respectively.According to the Rat Brain Stereotaxic Atlas, 5 μL of aggregation of Aβ25-35 was injected into the right lateral ventricle to establish the AD rat model.7 days after successful modeling, Morris water maze was used to test thechanges of learning and memory ability of the rats.Results There was no significant difference in the average swimming speed between the two groups (P > 0.05).The escape latency time of rats in the model group was significantly increasedcompared with the sham group (P < 0.05).In the model group, the escape latency time of rats treated with 4 μg/μL and 8 μg/μL Aβ25-35 was significantly increased compared with the rats injected with 2 μg/μL (P < 0.05), while there was no significant difference between rats treated with 4 μg/μL and 8 μg/μL Aβ25-35 (P > 0.05).The activity time and distance of target quadrant of the rats injected with different concentration of Aβ25-35in the model group were significantly reduced compared with the sham group (P < 0.05), but no significant difference amongthe rats treated with different Aβ25-35 concentrations (P > 0.05).Compared with the sham-operated group, the number of platform-crossing of rats injected with different doses of Aβ25-35in the model group were significantly reduced (P < 0.05).In the model group, the rats treated with 4 μg/μL and 8 μg/μL was significantly reduced compared with the group with 2 μg/μL injection (P < 0.05).There was no significant difference between the rats injected with 4 μg/μL and 8 μg/μL (P > 0.05).Conclusions The recommended dose and concentration of Aβ25-35 to be injected into the unilateral ventricle to establisha rat model of Alzheimer's disease is 4 μg/μL in a volume of 5 μL.
9.Acute renal failure in systemic lupus erythematosus
Ling ZHOU ; Li-Jun SUN ; Jun BAO ; Zhen XU ; Hu-Ji XU ;
Chinese Journal of Rheumatology 2003;0(07):-
Objective To understand the clinical features of acute renal failure(ARF)as the initial presentation of systemic lupus erythematosus(SLE).Methods Eight cases of ARF in SLE from Jan 1995 to Apt 2006 were investigated,descriptive analysis and literature review were performed.Results①The symp- tom of ARF in SLE was mainly oliguria,with severe accompany symptoms and complications.②The level of leucocyte and hemoglobin was low in laboratory tests,also the complement level decreased significantly.The most frequent renal pathology was typeⅣ,Ⅳ+ⅤLN.③Large dose steroid and CTX were the mainstay of treatment.In addition,SCUF,CVVHDF and hemodialysis could be used for lethal conditions.Conclusion ARF can be the first manifestation of SLE and it usually represents more severe disease and more complica- tions.Large dose steroid and CTX can improve prognois.In cases refractory to steroid and if the effect is obso- lete,CTX treatment SCUF,CVVHDF and hemodialysis can be use.
10.Clinical and molecular genetic analysis of a multigenerational pedigree with generalized epilepsy with febrile seizures plus
Shui-Zhen ZHOU ; Yi-Qing SONG ; Chao CHEN ; Dao-Kai SUN ;
Chinese Journal of Neurology 2005;0(07):-
-2,and T)was detected on exon 11 in the mutational analysis of GABRG2.Our results indicate that genomic variations of GABRG2 are not likely to be substantially involved in the etiology of GEFS+in this family. Conclusion Our study fails to provide evidence supporting a causal relation between the SCN1A,SCN1B, GABRG2 mutation and the etiologic genes in this family,which indicates that GEFS+has with phenotypic and genotypic heterogeneity.