2.Establishing a serum-free primary culture method for cortical neurons of new-born rats
Chuanhao YANG ; Dong ZHAO ; Qi LIU ; Yunxiang JI ; Lingjian LI ; Hui XU ; Jing DAI ; Xuejun HE ; Yezhong WANG
Chongqing Medicine 2014;(29):3901-3903,3906
Objective To establish a serum-free primary culture method for cortical neurons of new-born rats .Methods The cortical tissue was digested and the cells were planted in the medium containing 10% fetal bovine serum ,and then maintained feed-ing with neurobasal medium containing B27 after 4 to 8 h .The morphology was observed under phase-contrast microscope .RT-PCR ,Western blot and immunocytochemistry were applied to identify the expression of NSE gene and protein in neurons .Results A large number of neurons began to adhere to the cover glasses after 2 to 8 h .They showed different shapes-shuttle ,triangle pyram-idal ,or no regular after clinging to the plate .Their processes connected to nets and were different in length and thickness .They well developed at the 7th to 10th day .The isolated and cultured cells were confirmed as neurons by RT-PCR ,Western blot and immuno-cytochemistry .Conclusion This technique is an easy and practical tool for primary culture of new-born rats cortical neurons with high purity ,and can be used as an in-vitro model of research .
3.Effects of herbal compound 861 on hepatic stellate cell expressing endothelin-1 protein and mRNA.
Hui-guo DING ; Shu-zhen TANG ; Bao-en WANG ; Ji-dong JIA ; Chun-hui ZHAO
Chinese Journal of Hepatology 2003;11(5):308-308
Animals
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Cells, Cultured
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Drugs, Chinese Herbal
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pharmacology
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Endothelin-1
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biosynthesis
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genetics
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Hepatocytes
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metabolism
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Liver
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cytology
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metabolism
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Liver Cirrhosis
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prevention & control
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RNA, Messenger
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biosynthesis
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genetics
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Rats
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Rats, Sprague-Dawley
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Simian virus 40
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genetics
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Transfection
4.Incidence of vascular anomalies and variants associated with unilateral venous pulsatile tinnitus in 242 patients based on dual-phase contrast-enhanced computed tomography.
Cheng DONG ; Peng-Fei ZHAO ; Ji-Gang YANG ; Zhao-Hui LIU ; Zhen-Chang WANG
Chinese Medical Journal 2015;128(5):581-585
BACKGROUNDA comprehensive assessment of various vascular anomalies and variants associated with venous pulsatile tinnitus (PT) by radiography is essential for therapeutic planning and improving the clinical outcome. This study evaluated the incidence of various vascular anomalies and variants on the PT side and determined whether these lesions occurred as multiple or single entities.
METHODSThe dual-phase contrast-enhanced computed tomography images of 242 patients with unilateral venous PT were retrospectively reviewed. The vascular anomalies and variants on the symptomatic and asymptomatic sides were analyzed, and the incidences of anomalies or variants on each side were compared. The number of anomalies and variants on the symptomatic side in each patient was calculated.
RESULTS(1) A total 170 patients (170/242) had more than one anomaly or variant on the symptomatic side, and 58 patients (58/242) had a single lesion on tomography. (2) There was a statistically significant difference in the incidence of dehiscent sigmoid plate (P = 0.000), lateral sinus stenosis (P = 0.014), high jugular bulb (P = 0.000), sigmoid sinus diverticulum (P = 0.000), jugular bulb diverticulum (P = 0.000), dehiscent jugular bulb (P = 0.000), and a large emissary vein (P = 0.006) between the symptomatic and asymptomatic sides. (3) Dehiscent sigmoid plate (86.4%) was the most frequent lesion on the symptomatic side, followed by lateral sinus stenosis (55.8%), high jugular bulb (47.1%), sigmoid sinus diverticulum (34.3%), jugular bulb diverticulum (13.6%), dehiscent jugular bulb (13.6%), large emissary vein (4.1%), sinus thrombosis (1.2%), and petrosquamosal sinus (0.8%).
CONCLUSIONSVarious vascular anomalies and variants occur more frequently on the venous PT side. Preliminary findings suggest that venous PT patients may have multiple vascular anomalies or variants on the symptomatic side.
Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Tinnitus ; diagnostic imaging ; epidemiology ; etiology ; Tomography, X-Ray Computed ; methods ; Young Adult
5.Retreatment with interferon of relapsed chronic hepatitis C patients.
Ming-hui LI ; Yao XIE ; Hui ZHAO ; Wei-ni OU ; Dao-zhen XU ; Zhi-meng LU ; Kang-xian LUO ; Ji-dong JIA ; Yu-ming WANG ; Gui-zhen ZHAO ; Shu-lin ZHANG ; Da-zhi ZHANG
Chinese Journal of Hepatology 2006;14(1):3-6
OBJECTIVETo evaluate the efficacy and investigate the influencing factors of the interferon (IFN) retreatment for patients with chronic hepatitis C relapsed after a previous IFN treatment.
METHODSA retrospective study was designed to analyze the retreatment with IFN of 60 relapsed chronic hepatitis C patients. All patients were from a randomized, opened and multi-center clinical trial about the efficacy and security of PEG-IFNalpha-2a compared to CIFNalpha-2a in the treatment of chronic hepatitis C in China. There were 35 patients treated with PEG-IFNalpha-2a and 25 with CIFNalpha-2a. The main parameter to evaluate the efficacy was sustained viral response (SVR) rate. The influence of viral concentration in serum, genotype and drug categories on the responses to IFN were analyzed.
RESULTSFor all the patients, the end of treatment virus response (ETVR) and SVR rates were 55.00% and 35.00% respectively. ETVR rate of PEG-IFNalpha-2a was significantly higher than that of CIFNalpha-2a (74.29% and 28.00% respectively, P < 0.01). SVR rate of PEG-IFNalpha-2a was also markedly higher than that of CIFNalpha-2a (45.71% and 20.00% respectively, P < 0.05). However, there was no significant difference between the high and low viral load groups. Among the patients with genotype 1, ETVR and SVR rates of PEG-IFNalpha-2a (75.00%, 45.83%) were significantly higher than those of CIFNalpha-2a (22.22%, 11.11%), (P < 0.01, P < 0.05 respectively), but in patients with genotype non-1, there were no such differences between the two groups.
CONCLUSIONSome relapsed patients were not responsive to the IFN retreatment. The efficacy of PEG-IFNalpha-2a was superior to CIFNalpha-2a. The conventional IFN was not suggested to be used in the relapsed cases with genotype 1. The viral load was not associated with the efficacy of IFN retreatment.
Adult ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis C, Chronic ; therapy ; Humans ; Interferon-alpha ; therapeutic use ; Interferon-beta ; Interferons ; therapeutic use ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; Recurrence ; Retrospective Studies
6.The factors associated with viral relapse after interferon treatment in chronic hepatitis C patients.
Lie MA ; Hui ZHAO ; Yao XIE ; Ming-hui LI ; Dao-zhen XU ; Zhi-meng LU ; Kang-xian LUO ; Ji-dong JIA ; Yu-ming WANG ; Gui-zhen ZHAO ; Shu-lin ZHANG ; Da-zhi ZHANG
Chinese Journal of Hepatology 2006;14(8):565-568
OBJECTIVETo investigate the relationship between hepatitis C virus (HCV) genotype, serum viral load and ALT levels, and the factors associated with the viral relapse after IFN treatment in patients with chronic hepatitis C.
METHODSThe HCV RNA levels were determined with Cobas Amplicor Monitor Test, version 2.0, and HCV genotypes were examined by means of PCR products of 5' NTR digested with restriction endonucleases. The patients with chronic hepatitis C were treated with PEG-IFN alpha -2a and Roferon-A for 24 weeks. Those with a viral response after 24 week treatment were followed for an additional 24 weeks. The association of clinical characteristics, such as sex, age, the way of the HCV infection, IFN treatment history and platelet counts, and the HCV genotype, virus load and medicine used for the viral relapse after IFN treatment were analyzed.
RESULTSOf the 208 chronic hepatitis C patients, the ALT levels were not related to HCV RNA levels (r = 0.093, P > 0.05). No difference of ALT levels between HCV genotypes was found, and the HCV RNA load was also of no difference between HCV genotype 1 patients and non 1 patients. Of the 119 patients with viral response after 24 week treatment, 58 cases (48.7%) relapsed after another 24 week's follow-up. Relapse was not significantly related to the clinical characteristics, such as sex, age, mode of the infection, treatment history of IFN, AST/ALT ratio, platelet counts and the baseline viral load. Among patients with genotype 1 virus, the relapse rate was significantly higher than those patients with non-genotype 1 virus (54.5% vs 32.1%, P=0.039). The relapse rate after PEG-IFN alpha -2a treatment was lower than that of Roferon-A treatment (47.0% vs. 52.8%), but not significantly.
CONCLUSIONThe viral relapse of chronic hepatitis C patients after IFN treatment was significantly associated with the genotypes of the HCV.
Antiviral Agents ; therapeutic use ; Female ; Genotype ; Hepacivirus ; genetics ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Middle Aged ; RNA, Viral ; blood ; Recombinant Proteins ; Recurrence ; Treatment Outcome ; Viral Load
7.Non-relationship between HCV genotype,serum viral load,and ALT,AST levels in chronic hepatitis C patients
Hui ZHAO ; Yao XIE ; Ming-Hui LI ; Dao-Zhen XU ; Zhi-Meng LU ; Kang-Xian LUO ; Ji-Dong JIA ; Yu-Ming WANG ; Gui-Zhen ZHAO ; Shulin ZHANG ; Dazhi ZHANG ;
Chinese Journal of Laboratory Medicine 2000;0(06):-
0.05),the difference of ALT,AST level and AST/ALT ratio between high viral load (serum HCV RNA≥8.5?10~5 IU/ml) group and low viral load (serum HCV RNA
8.Aminoguanidine alleviates early brain injury after subarachnoid hemorrhage in rats
Peng YANG ; Dong ZHAO ; Qi LIU ; Yunxiang JI ; Licang ZHU ; Jing DAI ; Hui XU ; Shijun TANG ; Wenxue ZHU ; Xiaotian LI ; Yezhong WANG
Journal of Medical Postgraduates 2015;(8):794-798
Objective Recent studies have indicated that early brain injury is the leading cause of death in patients with subarachnoid hemorrhage ( SAH) .Our study investigated the role of aminoguanidine ( AG) in early brain injury after SAH . Methods Sixty-eight male SD rats were equally randomized into four groups of equal number :control, sham, SAH, and AG.The animals in the sham group were injected with isotonic saline solution , while those of the latter two groups with femoral artery blood ( FAB) and FAB+AG, respectively, into the pre-chiasmatic cistern to induce SAH. At 24 hours after modeling , all the rats were killed for HE staining , obtainment of behavioral neurological assessment ( BNA ) scores by Garcia, measurement of the apoptosis of neurons by TUNEL , and de-termination of the expressions of the iNOS and NSE proteins by West-ern blot. Results The results of HE staining showed the presence of more red blood cells in the subarachnoid cavity of the rats in the SAH group, with a significantly decreased BNA score ( 14.47 ± 0.62) as compared with those in the control (17.94 ±0.24), sham (17.59 ±0.51), and AG group (15.71 ±0.47) (P<0.05). The rate of positive cells was remarkably higher in the SAH group ([42.38 ±2.38]%) than in the control ([6.35 ±0.94]%), sham ([6.85 ±0.69]%), and AG group ([30.48 ±2.89]%) ( P<0.01), with significant differences among the latter three groups (P<0.05).The expressions of iNOS and NSE were markedly higher in the SAH group ([3.86 ±0.07] and [1.59 ±0.06]) than in the control (0 and[0.35 ±0.09]), sham ([2.96 ±0.34] and [0.38 ±0.08]), and AG group ([3.41 ±0.04] and [0.70 ±0.12]) ( P<0.05).Both the expression levels of iNOS and NSE were positively correlated with the rate of positive cells (r=0 .879 and 0.935, P<0.01). Conclusion AG can alleviate early brain injury after SAH in SD rats by improving the neuro-ethologic function , suppressing the apoptosis of neurons , and reducing the expressions of iNOS and NSE .
9.Clinical characteristics comparison between triple-negative and non-triple-negative breast cancer.
Hui-ming ZHANG ; Li-xue XUAN ; Ji-dong GAO ; Bao-ning ZHANG ; Ping ZHAO
Chinese Journal of Surgery 2009;47(7):506-510
OBJECTIVETo compare the clinical characteristics of triple-negative (TN) breast cancer and non-triple-negative (NTN) breast cancer, enrich the information of TN patients, and provide evidences for individualized combined treatment.
METHODSThe data of 408 cases received operation in the year of 2002 was enrolled in this study. TN patients were confirmed according to the immunohistochemical (IHC) test of estrogen receptor (ER), progesterone receptor (PR) and HER-2/neu. The clinical characteristics, recurrence, metastasis and survival were compared between the two groups.
RESULTSSeventy-seven patients (18.9%) were confirmed TN cases. The median follow-up was 64 months (range, 3-79 months). Of all the cases, 58 occurred local recurrence or metastasis and 51 died, it was 19 and 12 in TN group. Compared with the NTN group, the TN patient tended to be younger and the tumor mass larger (P=0.015 and 0.011). However, axillary lymph nodes metastasis occurred more often in NTN patients than in TN patients (P=0.001). The rate of local recurrence and metastasis in TN group was significantly higher than in NTN group (P=0.005 and 0.025), and TN cases were more likely to develop lung metastasis than NTN patients (P<0.01). The 3-year and 5-year overall survival rate in TN group were significantly lower than in NTN group (86.4% vs. 93.4%, P=0.0205; 77.7% vs. 87.9%, P=0.0215). The 3-year and 5-year disease-free survival rate in TN group were also significantly lower than in NTN group (78.4% vs. 92.4%, P=0.0038; 72.8% vs. 85.8%, P=0.0041). Tumor size, lymph node status and triple-negative were the most important factors influencing the prognosis on multivariate Cox regression analysis.
CONCLUSIONSTN breast cancer haa some specific clinical characteristics. The prognosis of TN patients is worse than that of NTN patients. Further study is needed to find individualized treatment for TN breast cancer patients.
Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; metabolism ; pathology ; therapy ; Female ; Follow-Up Studies ; Humans ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Prognosis ; Receptor, Epidermal Growth Factor ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Retrospective Studies ; Survival Analysis ; Young Adult
10.Establishment and preliminary characterization of hybridoma cell lines secreting monoclonal antibodies against Prion Proteins.
Li ZHAO ; Rong JI ; Jian-wei WANG ; Chun-hui HAN ; Xiu-ping YU ; Xiao-ping DONG ; Tao HUNG
Chinese Journal of Experimental and Clinical Virology 2003;17(2):133-136
OBJECTIVETo obtain monoclonal antibodies (McAbs) which can be widely used to detect mammalian prions (PrP) and to develop diagnostic tests for screening transmissile spongiform encephalopathies (TSE) as well as for studying pathogenesis of prion-related diseases.
METHODSBALB/c mice were immunized separately with bovine PrP peptide 29-48 (BoP1) and 89-108 (BoP2) coupled to keyhole limpt hemocyan. Two hybridoma cell lines secreting monoclonal antibodies against these peptides were established by cell fusion and 2 to 3 rounds of cell cloning. The reactions of the McAbs to the recombinant bovine (Bo)PrP(25-242), human (Hu)PrP(23-231) and hamster (Ha) PrP (23?231) were tested separately by Western blotting.
RESULTSThrough cell fusion, two hybridoma cell lines secreting McAbs against BoP1 and BoP2, designated D11 and D8 accordingly, were identified by ELISA and cell cloning. The McAbs produced by these cell lines reacted well with the recombinant PrP proteins; (Bo) PrP (25-242), (Hu) PrP (23-231), and (Ha) PrP (23-231), respectively.
CONCLUSIONSTwo McAbs reacting with bovine, human and hamster PrPs were successfully generated, they are potential to be used to detect PrPs in mammals and to study the mechanism of pathogenesis of TSE.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Antibodies, Viral ; biosynthesis ; immunology ; Antibody Specificity ; Cattle ; Cricetinae ; Cross Reactions ; Encephalopathy, Bovine Spongiform ; prevention & control ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Hybridomas ; secretion ; Male ; Mice ; Mice, Inbred BALB C ; PrPSc Proteins ; immunology ; Prion Diseases ; prevention & control ; Prions ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; immunology