1.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.
2.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
3.Effect of oxymatrine on mouse allergic contact dermatitis induced by DNFB and lymphocyte proliferation stimulated by Con A
Bin WU ; Xiaochang CAI ; Yaoying ZENG ; Jun SHI ; Tong WANG ; Jingxian ZHAO ; Xiuyan HUANG
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To explore the inhibitory effect of oxymatrine (OMT) on the allergic contact dermatitis (ACD) stimulated by dinitrofluorobenzene (DNFB) and its effects on the proliferation of the lymphocytes. METHODS: ① An ACD mouse model was established by stimulation with DNFB, and then the mice were injected intraperitoneally with different dosages of OMT, PBS and hydrocortisone (HCT) respectively, the swelling degree of their auricles was examined. ② Carboxyfluorescein diacetate, succinimidyl ester (CFDA-SE) dye and flow cytometer were used to examine the fluorescence intensity changes of lymphocytes stimulated by polyclonal stimulator ConA and OMT. RESULTS: ① compared with PBS group, OMT possessed the strong inhibitory effect on the ACD caused by DNFB in a dose-dependent manner, and its inhibitory effect was equivalent to the HCT of the same dosage with fewer side effects. ② In vitro experiments proved that OMT (500, 125 and 31 mg/L) had the ability to restrain the proliferation of lymphocytes of mouse. CONCLUSION: OMT possesses an inhibitory effect on the ACD induced by DNFB, and OMT is a kind of immunosuppressor.
4.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.
5.Construction and immunological responses of recombinant adenovirus containing Epstein-Barr nuclear antigen 1 in mice.
Yan-Yan TONG ; Hong-Xia LI ; Li-Xia ZHANG ; Zhan WANG ; Ling ZHOU ; Yi ZENG ; Hai-Jun DU
Chinese Journal of Virology 2014;30(4):429-435
This study aimed to construct recombinant adenovirus expressing Epstein-Barr nuclear antigen 1 (EBNA1) against nasopharyngeal carcinoma (NPC). The C-terminal region fragment of the ebna1 gene of Epstein-Barr virus was amplified from the standard strain B95-8 by polymerase chain reaction (PCR). The gene fragment was inserted into the pDC316 shuttle plasmid using the EcoRI and BgIII restriction enzyme sites. The pDC316-ebna1 shuttle plasmid and pBHG helper plasmid were cotransfected into HEK293 cells after sequencing. The soluble protein was extracted from HEK293 cells, which caused apparent cytopathic effects. The transcription and expression of the ebna1 gene were confirmed using flow cytometry and Western blotting. rAd-ebna1 titers were measured by the TCID50. rAd-ebna1 was injected into BALB/c mice at a dose of 2 x 10(8) VP per mouse, EBNA1 epitope-specific responses were measured at 1st, 2nd, 4th and 8th weeks post-immunization. The target fragment of ebna1 (939 bp) was obtained by PCR, and was in consensus with the sequence from the standard strain B95-8. Cytopathic effects were observed after the pDC316-ebna1 shuttle plasmid and pBHG helper plasmid were cotransfected into HEK293 cells. rAd-ebna1 was successfully recombined in HEK293 cells. EBNA1 protein was detected in HEK293 cells, rAd-ebna1 titers reached 10(8) TCID50/mL. Specific responses to CD4+ epitopes of EBNA1 were detected in the immunized mice. In conclusion, rAd-ebna1 was successfully constructed and induced specific responses to CD4+ epitopes of EBNA1 in immunized mice.
Adenoviridae
;
genetics
;
immunology
;
Animals
;
CD4-Positive T-Lymphocytes
;
immunology
;
virology
;
Epstein-Barr Virus Infections
;
immunology
;
prevention & control
;
virology
;
Epstein-Barr Virus Nuclear Antigens
;
administration & dosage
;
genetics
;
immunology
;
Genetic Vectors
;
genetics
;
immunology
;
Herpesvirus 4, Human
;
genetics
;
immunology
;
Humans
;
Male
;
Mice
;
Mice, Inbred BALB C
;
Viral Proteins
;
administration & dosage
;
genetics
;
immunology
6.Enatiomeric separation of beta-blocking agents and analogs
Lu-Shan YU ; Tong-Wei YAO ; Xiang-Jun WANG ; Su ZENG
Journal of Zhejiang University. Medical sciences 2002;31(6):414-418
OBJECTIVE: To evaluate enantiomeric separation methods for beta-blocking agents and analogs. METHODS: Enantiomeric separation of racemates of 11 beta-blocking agents and their analogs was performed using chiral stationary phases and 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl isothiocyanate (GITC). RESULTS: These beta -blocker racemates were separated into enantiomers in one or several chormatographic states such as propranolol, bisoprolol, metoprolol, celiprolol, carvedilol, sotalol, propafenone, ephedrine, and zomitriptan. Temperature had a significant effect on the resolution of the drugs when using chiralcel OD. Lower temperatures were associated with higher resolutions. CONCLUSION: When separating beta-blocking agents and their analogs, Chiralcel OD, Chiralpak AD, Chiral stationary phases and GITC chiral derivative reagents have complementary functions.
8.Effects of Tongyuan acupuncture combined with salt and moxibustion Shenque on behavior and hypotha-lamic-pituitary-adrenal axis in postpartum depression rats
Guifeng ZHANG ; Xiaoling WU ; Tong-Jun ZENG ; Yushan YU ; Yiwen ZENG ; Liqiang LI
The Journal of Practical Medicine 2018;34(8):1262-1266
Objective To explore the effects of Tongyuan acupuncture combined with herbal medicine and moxibustion on the behavior and hypothalamic-pituitary-adrenal axis-related hormone levels in postpartum de-pression rats.To explore the therapeutic mechanism of this method in the treatment of postpartum depression.Meth-ods A total of 120 SD rats were randomly divided into 6 groups(n=20).The blank group was not treated.The sham operation group was performed without surgery to remove the ovaries. The model group,drug group,routine acupuncture group and experimental group were established in the model of postpartum depression.After modeling successfully,the model group did not interfere with any treatment. The rats in treatment group were treated with fluoxetine hydrochloride. The acupuncture group was treated with acupuncture of Baihui and Taichong. The rats in the experimental group were treated with the method of acupuncture and moxibustion.At the end of the second and fourth week after the initiation of intervention,10 rats were randomly selected from each group.They were sacrificed and the levels of serum corticotropin-releasing hormone(CRH),adrenocorticotropic hormone(ACTH),cortisol (Cor)were measured.Results The levels of CRH,ACTH and Cor in the model group were higher than those in the blank group(P<0.05).The levels of serum CRH,ACTH and Cor in the drug group,routine acupuncture group and experimental group were lower than those in the model group(P<0.05).The levels of serum ACTH and Cor in the experimental group were lower than those in the drug group and routine acupuncture group(P<0.05).There was no significant difference in serum CRH,ACTH and Cor level between the experimental group and blank group(P >0.05).Conclusions Tongyuan acupuncture combined with salt moxibustion and Shenque have significant effects on the depressive symptoms and serum levels of CRH,ACTH and Cor in postpartum depression rats,which may be re-lated to the regulation of hypothalamic-pituitary-adrenal axis-related hormone levels.
9.Multivariate analysis of recurrence in T1 bladder transitional cell carcinoma.
Jun ZHANG ; Li ZENG ; Gui-zhong LI ; Xin-yu YANG ; Tong-li XIA ; Yan-qun NA ; Ying-lu GUO
Chinese Journal of Oncology 2003;25(5):475-477
OBJECTIVETo investigate the prognostic value of some clinicopathologic indexes and biologic tumor markers in predicting recurrence in T1 transitional cell carcinoma (TCC) of the bladder.
METHODSThe expressions of p53, E-cadherin and VEGF of 75 patients with T1 primary bladder TCC were detected by streptabitin peroxidase (SP) immunohistochemical methods. The effects of clinicopathologic indexes and biologic tumor markers on recurrence were assessed by Kaplan-Meier and Cox proportional hazards model.
RESULTSThe 1-, 3-, and 5-year recurrence- free survival rates were 68.0%, 45.3% and 20.9%. In Kaplan-Meier analysis, tumor mutifocality and the expression of p53, E-cadherin and VEGF were associated with recurrence. In multivariate analysis, the independent recurrence variables were tumor mutifocality, the expression of p53 and E-cadherin.
CONCLUSIONTumor mutifocality and the abnormal expression of p53 and E-cadherin are the variables that independently predict recurrence in T1 transitional cell carcinoma of the bladder.
Adult ; Aged ; Aged, 80 and over ; Cadherins ; analysis ; Carcinoma, Transitional Cell ; etiology ; mortality ; Female ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Recurrence, Local ; etiology ; mortality ; Neoplasm Staging ; Survival Rate ; Tumor Suppressor Protein p53 ; analysis ; Urinary Bladder Neoplasms ; etiology ; mortality ; Vascular Endothelial Growth Factor A ; analysis
10.Effects of blocking androgen receptor expression with specific hammerhead ribozyme on in vitro growth of prostate cancer cell line.
Qiangsong TONG ; Jun ZHAO ; Zhaohui CHEN ; Fuqing ZENG ; Gongcheng LU
Chinese Medical Journal 2003;116(10):1515-1518
OBJECTIVETo study the possibility of gene therapy for prostate cancer by blocking androgen receptor (AR) gene expression using a specific hammerhead ribozyme (RZ).
METHODSThe hammerhead ribozyme expression vector pcDNA-hAR-RZ, specific to AR mRNA, was constructed and transfected into the prostate cancer cell line LNCaP by using lipofectamine. Androgen receptor expression was measured by RT-PCR and immunohistochemical methods. Cellular proliferation activities were assayed using the tetrazolium bromide colorimetry method; cell cycle changes were observed by flow cytometry; and cell apoptosis was detected by the TdT-mediated dUTP-biotin nick end labeling method.
RESULTSOne to seven days after transfection with the ribozyme expression vector, AR mRNA expression at molecular and protein levels in LNCaP cells decreased by 32.6% - 40.7% (P < 0.05) and 21.0% - 87.64% (P < 0.05) respectively, and cell proliferation was inhibited by 18.28% - 35.34% (P < 0.05). Meanwhile, the cell cycle was arrested at the G2/M stage, and apoptotic morphological changes occurred with an apoptosis rate of 25.17% (P < 0.01).
CONCLUSIONRibozyme specific against AR mRNA is capable of inhibiting the expression AR and inducing the apoptosis in prostate cancer cells.
Apoptosis ; Cell Line, Tumor ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Genetic Vectors ; Humans ; Immunohistochemistry ; Male ; Prostatic Neoplasms ; therapy ; RNA, Catalytic ; physiology ; Receptors, Androgen ; analysis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection