1.Effects of recombinant hirudin variant Ⅲ on expression of apoptosis-related genes during galactose-mediated human lens epithelial cells damage
Yu, OU ; Zhi-jun, YUAN ; Pei, GENG ; Wu-tong, WU
Chinese Journal of Experimental Ophthalmology 2011;29(7):581-584
Background Recombinant hirudin variant Ⅲ(rHV3) can effectively prevent galactose-induced human lens epithelial cells LECs injury,but little is known about the molecular mechanism of its action.Objective The present study was to investigate the effects of rHV3 on the expression of apoptosis-related genes in damaged LECs induced by galactose.Methods The rHV3 was extracted by our research group,and the biological activity of rHV3 was identified by titration of thrombase according to Markwardt's method.Human LECs (SRA01/04) were cultured using 125×10-3 mol/L D-galactose+10% FBS+D/F12 medium to establish the damaged human LECs model.rHV3 was added into the medium of the damaged human LECs model.Human LECs were cultured in D/F12 medium containing 10% FBS as normal control.The expression of apoptosis-related genes,such as aldose reductase (AR),bax,bcl2 and p53,in LECs at the mRNA level was detected using RT-PCR.The abundance ratio of target genes was presented with the absorbance (A) of gene mRNA/GAPDH mRNA.Results Compared to the normal control group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were significantly elevated in model group (t=3.90E-06,t=8.44E-04,t=5.15E-08,P<0.01).However,in the rHV3-treated group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were lower than those of model group (t=5.90E-06,t=1.51E-04,t=3.42E-06,P<0.01).The bcl2 mRNA/GAPDH mRNA was markedly downregulated in the model group when compared with the normal control group (t=1.86E-05,P<0.01);while after rHV3 addition,bcl2 mRNA/GAPDH mRNA increased in comparison with the model group (t=8.56E-05,P<0.01).Conclusion 125×10-3mol/L D-galactose induces the damage and apoptosis of human LECs.rHV3 likely plays a protective function on D-galactose-induced damage of human LECs by inhibiting the polyol pathway and mitochondria-mediated pathway.
2.The clinical and genetical characteristic of one dentatorubral-pallidoluysian atrophy pedigree with an onset of cognitive impairment
Juanjuan CHEN ; Zhenxing ZENG ; Jun WU ; Xiaoxin TONG ; Li YI
Chinese Journal of Neurology 2013;46(10):692-696
Objective To report the clinical and genetic characteristics of a dentatorubralpallidoluysian atrophy (DRPLA) pedigree with an onset of cognitive impairment.Methods Clinical data of this pedigree was collected.The numbers of CAG repeats in the exon 5 of atrophin-1 (ATN1) gene were analysed in the proband and the other 4 healthy family individuals.The polymerase chain reaction (PCR) products of the proband underwent cloning-sequencing using an original TA cloning kit.Results There were 5 patients in this family,4 with onset in adult and one in childhood.The proband had an onset manifestation of cognitive impairment,while the other 3 adult patients presented with ataxia.The two-year-old child in the pedigree had myoclonic epilepsy.The proband had 61 CAG repeats in the exon 5 of ATN1 gene.After TA cloning-sequencing of the proband ' s PCR products,there were 2 different numbers of CAG repeats,including 61 and 64.Conclusions The clinical manifestations of DRPLA can have obvious heterogeneity in one family.Some patients present with cognitive impairment.It is very important to test the numbers of CAG repeats of ATN1 gen for DRPLA diagnosis.Somatic mosaicism may be also observed in Chinese DRPLA patients.
3.Clinical analysis of 39 cases with reoperation for thyroid
Han DENG ; Chuang TONG ; Jianguo WANG ; Jun CAO ; Zhiguo WU
Chinese Journal of Primary Medicine and Pharmacy 2014;(14):2099-2101
Objective To explore the cause of the thyroid reoperation ,methods,surgical approach ,and the prevention of complications .Methods 39 cases of thyroid reoperation in patients with clinical data were reviewed and analyzed.Results Thyroid papillary carcinoma in 11 cases,follicular carcinoma in 2 cases,nodular 22 cases of thyroid,thyroid adenoma 1 case,3 cases of primary hyperthyroidism .The incidence of transient hypoparathyroidism ac-counted for 2 cases,temporary laryngeal recurrent nerve injury in 2 cases,chyle leakage in 1 case,in total of 12.8%of postoperative complications .Conclusion Improper operation method choice for the first time and the misdiagnosis was the main cause of reoperation , The reoperation of thyroid is difficult and has more complications .Preoperative evaluation and careful operation can prevent the occurrence of complications .
4.Application of Intelligent Device for Energy Expenditure and Activity in Evaluation of Protrusion of Intervertebral Disc
Jianxian WU ; Jun HUANG ; Wei JIANG ; Bin WANG ; Tong LIU
Chinese Journal of Rehabilitation Theory and Practice 2008;14(3):260-262
Objective To apply the Intelligent Device for Energy Expenditure and Activity(IDEEA)system to evaluate the locomotion of patients with protrusion of intervertebral disc(PID)quantitatively and dynamically.Methods 8 patients with PID and 9 volunteers as controls performed a series of daily activities in their nature environment.They were monitored with IDEEA.Results The velocity of flexion forward,time flexion forward,time stand up,time sit down of the patients were significant different to the controls(P<0.05),and the walking speed of the former was slower(P<0.05)than the latter's.Conclusion Physical performance of the PID patients was significantly reduced.The IDEEA can be a good tool to evaluate their ability of locomotion in their daily living.
6.Study on regulatory effect of Danshensu on lipid metabolism of hyperlipidemia rats.
Juan CHEN ; Jun DENG ; Yu-yan ZHANG ; Jian-guo LI ; Fei-yu WU ; Hai-tong WAN
China Journal of Chinese Materia Medica 2015;40(2):313-317
OBJECTIVETo explore the effect of Danshensu on the lipid metabolism of hyperlipidemic rats.
METHODSixty clean male SD rats were selected. Twelve of them were selected in the basic control group and fed with common foods, and the remaining rats were fed with the high-fat feeds. After the successful modeling, they were randomly divided into the high-fat control group and low dose (10 mg x kg(-1) x d(-1)), medium dose (20 mg x kg(-1) x d(-1)) and high dose (40 mg x kg(-1) x d(-1)) Danshensu (dissolved in saline) groups. Both of the two groups were abdominally injected with the same volume of normal saline once a day for consecutively 30 days. The serum TG, TC, HDL-C and liver ACC1, FAS, HMGR, CPT-I mRNA expressions were detected.
RESULT AND CONCLUSIONDanshensu could inhibit the LDL-C level, timely clear redundant cholesterol and effectively regulate the lipid metablism of hyperlipidemic rats by reducing the TC content, decrease the fatty acid by reducing the FAS mRNA expression, and reduce the synthesis levels of endogenous cholesterol by inhibit the HMGR mRNA expression.
Animals ; Hyperlipidemias ; drug therapy ; metabolism ; Lactates ; pharmacology ; Lipid Metabolism ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley
7.Effects of rapamycin on amyloid β-protein induced impairments of working memory and synaptic plasticity in rats.
Ming HAO ; Jia-qing TONG ; Jun ZHANG ; Mei-na WU ; Jin-shun QI
Chinese Journal of Applied Physiology 2016;32(1):18-21
OBJECTIVEThe present study investigated the effects of rapamycin on Aβ1-42-induced deficits in working memory and synaptic plasticity.
METHODSAfter bilateral hippocampal injection of Aβ1-42 and rapamycinin rats, spontaneous alternation in Y-maze and in vivo hippocampal long-term potentiation (LTP) of rats were recorded. All data were analized by two-way repeated measures analysis of variance (ANOVA).
RESULTS(Hippocampal injection of Aβ1-42 alone impaired working memory of rats; (2) Rapamycin did not affect working memory of rats, but alleviated Aβ1-42-induced working memory deficits, compared with Aβ1-42 alone group; (Aβ1-42 remarkably suppressed in vivo hippocampal LTP of fEPSPs in the CA1 region; (4) Pretreatment with rapamycin prevented Aβ1-42-induced suppression of LTP.
CONCLUSIONThese data indicates that rapamycin could protect against Aβ1-42-induced impairments in working memory and synaptic plasticity in rats.
Amyloid beta-Peptides ; adverse effects ; Animals ; Hippocampus ; drug effects ; Long-Term Potentiation ; Maze Learning ; Memory, Short-Term ; drug effects ; Neuronal Plasticity ; drug effects ; Peptide Fragments ; adverse effects ; Rats ; Sirolimus ; pharmacology
8.Clinical, myopathological and genetic research of a patient with distal myopathy caused by caveolin-3 deficiency
Juanjuan CHEN ; Wenshuang ZENG ; Chunxi HAN ; Jun WU ; Xiaoxin TONG ; Haiou ZHANG
Chinese Journal of Neurology 2015;48(9):786-790
Objective To report the clinical,myopathological and genetic features of a patient with distal myopathy caused by caveolin-3 (CAV3) deficiency.Methods The patient was a 27-year-old female.She had an onset symptom of asymmetric lower extremities weakness.The proximal limb-girdle muscles were involved subsequently.Clinical data of this patient were collected.The leg muscle magnetic resonance imaging (MRI) and an open biopsy of left tibialis anterior muscle were performed.In addition to histological,enzyme histochemical staining and ultrastructural examination,immunohistochemical staining with antibody against CAV3 was done.CAV3 gene was analyzed in the patient and her parents.Results Tl-weighted enhanced skeletal muscle MRI of the lower limbs showed the abnormal signal in distal and proximal muscles.Muscle biopsy showed moderate dystrophic changes and immunostaining for CAV3 showed reduced plasmalemma in the muscle fibers.Gene analysis disclosed a heterozygous c.136G > A (p.Ala46Thr)mutation in the CAV3 gene,and the patient's parents did not have this mutation.Conclusions We report a distal myopathy case caused by c.136G > A (p.Ala46Thr) mutation in the CAV3 gene,who had an onset symptom of asymmetric lower extremities weakness.The proximal limb-girdal muscles were also involved.This would help clinical doctors to know more about this rare myopathy.
9.The study on the features of dopamine secretion in microencapsulated cells of bovine retinal pigmentary epithelial cells in vitro
Wang-Chun DU ; Jian WANG ; Zheng-Tong DING ; Jian-Jun WU ; Yuping JIANG ;
Chinese Journal of Geriatrics 2003;0(08):-
ObjectiveTo detect the features of dopamine secretion of cultured bovine retinal pigmentary epithelial(RPE) cells. The level of dopamine and survival rate after passage and microencapsulation were also observed. MethodsThe primary culture of bovine RPE cells was made using enzyme digestion. After purification and identification the growth curve of the cell was observed. Alginate-polylysine-alginate(APA) microencapsulated cells were made with a high voltage electostatic system. The activity of the cells in the mirocapsule was investigated by trypan blue staining. The secretion of dopamine was determined by high performance liquid chromatography (HPLC) assay. ResultsThe cell had high purity of immunocytochemistry. The growth curve showed that the exponential growth occurred at the first 1~4 days. Dopamine content was first detected at the time point of 2 hour, and arrived at the peak at about 48 hour of the cultivation. The secretion of dopamine was not different between passages. Dopamine secretion was dramatically decreased in the first 4 days after microencapsulation (P
10.Effects of oxymatrine on lymphocyte proliferation and the quantity of regulatory T cells
Bin WU ; Yaoying ZENG ; Xiaochang CAI ; Jun SHI ; Lin CONG ; Tong WANG ; Xiangfeng ZENG
Chinese Journal of Pathophysiology 2000;0(07):-
AIM:To analyze the effects of oxymatrine(OMT) on the quantity of murine regulatory T cells(Tr cells) in the peripheral blood and mouse lymphocyte proliferation stimulated by Con A,and to probe into the immunological mechanism that OMT treats allergic contact dermatitis(ACD).METHODS:An ACD mouse model stimulated by dinitrofluorobenzene(DNFB) was established.Different dosages of OMT,PBS and hydrocortisone(HCT) were intraperitoneally injected(IP) into the mice.Blood samples were collected at 1 d,7 d,14 d,21 d and 28 d,then the T cells were isolated and marked with anti-CD3,anti-CD4,anti-CD25 three-colored immune fluorescence antibody to detect the quantity of CD4+CD25+ T cells with flow cytometry.The fluorescence intensity changes of lymphocytes which were isolated from mouse's lymph node and co-stimulated by polyclonal stimulator Con A and OMT were examined by carboxyfluorescein diacetate succinimidyl ester(CFDA-SE) staining and flow cytometry.RESULTS:OMT at concentrations of 500,125 and 31 mg/L had the ability to restrain the proliferation of lymphocytes from lymph node in a dose dependent manner.However,OMT at concentrations of 16,8,4 and 2 mg/L promoted the proliferation of T lymphocytes from lymph node,but was not obviously dependent on its concentration.Intraperitoneal injection of OMT increased the numbers of CD4+CD25+T cell in peripheral blood obviously(P