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Sequencing of phosphopeptides sulfonated by 4-sulfophenyl isothiocyanate using post-source decay matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

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Progress in Natural Science:Materials International 2006年第9期16卷 930-935页

作者： TANG Jianguang , WANG Yongjun , ZHANG Hainan, WANG Chunyu, HU Zhiping, XUE Zhigang, XIA Kun3 and SHI Xiaoliu ( department of Neurology, the second xiangya hospital, central south university, changsha 410011, china department of digestion, the second xiangya hospital, central south university, changsha 410011, china National Key Laboratory of Chinese Genetics, changsha 410078, china) department of Neurology The Second Xiangya Hospital Central South University Changsha 410011 China department of digestion The Second Xiangya Hospital Central South University Changsha 410011 China National Key Laboratory of Chinese Genetics Changsha 410078 China

Phosphorylation/dephosphorylation is probably the most common and important reversible post-translational modification of proteins. Analyzing the functional effects of phosphorylation is helpful for understanding the biological functions of proteins. Identification of the phosphorylation sites of phosphorylated protein is a prerequisite for research on phosphorylation. In this work, an effective and simple method of identification of protein phosphorylation sites has been developed. Phosphopeptides were selectively enriched with immobilized metal affinity chromatography (IMAC) and subsequently chemically modified by 4-sulfophenyI isothiocyanate, and then the chemically modified phosphopeptides were sequenced with post-source decay (PSD) matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry for detecting phosphorylation sites. The charge of derivatization by 4-sulfophenyl isothiocyanate introduces a negative sulfonic acid group at the N-terminus of a peptide, and enables the selective detection of only a single series of C-terminal y-type ions. This chemically assisted method greatly simplifies the extremely complex pattern of PSD fragment ions and makes the PSD spectra more easier to be interpreted. The phosphorylation sites of a synthesized model phosphopeptide and human c-myc protein have been successfully identified by this method.

关键词： phosphopeptides sequencing 4-sulfophenyl isothiocyanate post-source decay matrix-assisted laser desorption/ion-ization time-of-flight mass spectrometry.

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Vitrectomy for vitreous amyloidosis

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International Journal of Ophthalmology(English edition) 2011年第3期4卷 307-310页

作者： Jiang You department of Ophthalmology,the second xiangya hospital of central south university,changsha 410011,Hunan Province,china department of Ophthalmology the Second Xiangya Hospital of Central South University Changsha 410011 Hunan Province China.

AIM: To study the clinical features of vitreous amyloidosis and the effect of vitrectomy for it. METHODS: We treated 6 eyes (4 Patients) with vitreous amyloidosis from 2008 to 2009, and followed up for 18-30 months after vitrectomy. RESULTS: The visual acuity ranged from counting fingers to 3/50 before surgery and 15/50-40/50 after vitrectomy. No severe complication was observed. CONCLUSION: Vitrectomy is an effective and safe treatment for eyes with vitreous amyloidosis. A long period of follow-up after surgery should be performed.

关键词： vitreous amyloidosis surgical treatment

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Potential role of retina as a biomarker for progression of Parkinson's disease

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International Journal of Ophthalmology(English edition) 2011年第4期4卷 433-438页

作者： Tian Tian 1, Xiao-Hua Zhu 2, Yun-Hai Liu 1 1department of Neurology, xiangya hospital, central-south university, 87 xiangya Road, changsha 410008, Hunan Province, china 2department of Ophthalmology, the second xiangya hospital, central-south university, 139 Renmin Middle Road, changsha 410011, Hunan Province, china department of Neurology Xiangya Hospital Central-South University 87 Xiangya Road Changsha 410008 Hunan Province China.

Optical coherence tomography (OCT) noninvasively quantifies the thickness of the retinal nerve fiber layer (RNFL). OCT has been studied in several neuro-ophthalmic conditions, including Parkinson’s disease (PD). Recent studies suggest that the quantitative analysis of RNFL can be precisely and noninvasively done by OCT scans and the results suggest that the thickness of RNFL is significantly decreased in patients with PD compared with age-matched controls and the foveal retinal thickness correlates with disease severity in PD. In this article, the application of OCT imaging of the retina in PD was reviewed. Literature survey of PubMed was carried out using the search terms of "Optical Coherence Tomography" combined with "Parkinson’s Disease" and "retinal nerve fiber layer" (without restriction to the year of publication). Some related articles were also included. The search was completed in Jul. 2011 and revised and updated as necessary. The aim of this article is to review the current literatures on the use of optical coherence tomography in patients affected by PD and to enhance its use in clinical practice in neuro-ophthalmology.

关键词： Parkinson disease optical coherence tomography retinal nerve fiber layer thickness

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Paclitaxel ameliorates fibrosis in hepatic stellate cells via inhibition of TGF-β/Smad activity

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World Journal of Gastroenterology 2010年第26期16卷 3330-3334页

作者： Jun Zhou, De-Wu Zhong, Qun-Wei Wang, Xiong-Ying Miao, Xun-Di Xu, department of Hepatobiliary Surgery, second xiangya hospital, central south university, changsha 410011, Hunan Province, china department of Hepatobiliary Surgery Second Xiangya Hospital Central South University Changsha 410011 Hunan Province China.

AIM: To investigated if paclitaxel can attenuate hepatic fi brosis in rat hepatic stellate cells (RHSCs). METHODS: RHSCs were cultured in vitro and randomly assigned to four groups: normal control group (treated only with Dulbecco's Modified Eagle's Medium), Taxol group (200 nmol/L paclitaxel was added to the cell culture), transforming growth factor (TGF)-β group (5 ng/mL recombinant human TGF-β1 was added to the cell culture), and TGF-β + Taxol group. TGF-β signaling cascade and status of various extracellular matrix proteins were evaluated by real time reverse transcriptase polymerase chain reaction and Western blotting. RESULTS: The paclitaxel treatment markedly suppressed Smad2/3 phosphorylation. This was associated with attenuated expression of collagen Ⅰ and Ⅲ and fi bronectin in ***: These data indicate that 200 nmol/L paclitaxel ameliorates hepatic fi brosis via modulating TGF-β signaling, and that paclitaxel may have some therapeutic value in humans with hepatic fi brosis.

关键词： Transforming growth factor-β Hepatic fibrosis Paclitaxel Smad Microtubules

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Multiple dose pharmacokinetics of risperidone and 9-hydroxyrisperidone in Chinese female patients with schizophrenia

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Acta Pharmacologica Sinica 2006年第3期27卷 381-386页

作者： Zhi-ling ZHOU~2 Xin LI~3 Huai-yan PENG~4 Xi-yong YU~2 Ming YANG~2 Feng-li SU~1 Feng WANG~1 Rong-hua ZHU~1 Chun-yu DENG~2 Qiu-xiong LIN~2 Chuan-yue WANG~5 Wen-biao LI~5 Shu-guang LIN~2 Huan-de LI~(1,6) ~1Clinical Pharmacy Research Institute,xiangya second hospital,central south university,changsha 410011,china ~2Research Center of Medical Science,Guangdong Provincial People's hospital,Guangzhou 510080,china ~3Vascular Surgery department,xiangya second hospital,central south university,changsha 410011,china ~4department of Clinical Laboratory,Third xiangya hospital,central south university,changsha 410013,china ~5Affiliated Beijing An Ding hospital,Capital university of Medical Science,Beijing 100088,china Research Center of Medical Science Guangdong Provincial People's Hospital Guangzhou China Vascular Surgery department Xiangya Second Hospital Central South University Changsha China department of Clinical Laboratory Third Xiangya Hospital Central South University Changsha China Clinical Pharmacy Research Institute Xiangya Second Hospital Central South University Changsha China Affiliated Beijing An Ding hospital Capital University of Medical Science Beijing China

Aim:To study the multiple dose clinical pharmacokinetics of risperidone and itsmain active metabolite,9-hydroxyrisperidone,in Chinese female patients ***:The subjects were 23 Chinese female inpatients aged18-65 years who met the CCMD-Ⅲ（third revision of the Chinese Criteria of Men-tal Disorders）criteria for *** were tested after 17 d of treat-ment with 2 mg risperidone twice *** concentrations of risperidone and9-hydroxy-risperidone were assayed by using validated high performance liquidchromatography-mass spectrometry（HPLC-MS）***:Risperidonewas rapidly absorbed(Tmaxwas 1.6 h)and its T1/2in plasma was short（3.2 h）.9-hydroxy-risperidone was quickly metabolized from the parent drug with a meanTmaxof 2.5 *** had a long half-life of 24.7 *** Cavssof risperidone and 9-hydroxy-risperidone were 36.9±33.1 and 110.6±30.5 μg·h·L-1,respectively,and the AUC0-12sswere 443.2+397.4 and 1327.2±402.3 μg·h·L-1,***/F and V/F ofrisperidone were 8.7±6.2 L/h and 34.1±24.3 L,*** varia-tions for pharmacokinetic parameters were quite large for *** 23 sub-jects experienced high prolactin levels when treated with *** was no correlation between prolactin level and the concentration ofrisperidone,9-hydroxy-risperidone,or the active ***:Risperidoneshowed large interindividual variations in *** ofrisperidone resulted in high serum prolactin *** results indicate that sys-temic exposure to risperidone and 9-hydroxy-risperidone in female Chinese schizo-phrenic patients is higher relative to published data for white Caucasian *** studies regarding the PK/PD relationship may be required to develop areasonable clinical dosage regimen for Chinese female patients.

关键词： pharmacokinetics risperidone 9-hydroxyrisperidone schizophrenia

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GPC3 fused to an alpha epitope of HBsAg acts as an immune target against hepatocellular carcinoma associated with hepatitis B virus

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Hepatobiliary & Pancreatic Diseases International 2011年第2期10卷 164-170页

作者： Jun-Wen Yang,Dong-Ye Yang,Fang-Gen Lu,Cai-Hong Li,Hui Chen,Ning Xie and Xin Zhao department of Digestive Diseases,second xiangya hospital,central south university,changsha 410011,china department of Digestive Diseases Second Xiangya Hospital Central South University Changsha 410011 China

BACKGROUND:The incidence of hepatocellular carcinoma (HCC)in china is closely related to the population infected with hepatitis B virus(HBV).HCC cells with HBV secrete soluble HBsAg into blood but do not express it on the cell membrane This study aimed to construct and investigate a new glycosyl phosphatidylinositol(GPI)-anchored protein(GPC3+α+EGFP) as a DNA vaccine against HCC associated with HBV. METHODS:A recombinant plasmid(pcDNA3.1(+)/GPC3+ α+EGFP)was constructed and verified by restriction endo nuclease digestion and ***3.1(+)/GPC3+α+ EGFP was transfected into HepG2 cells(experimental group) using lipofectamine ***-N1-transfected HepG2 cells were used as a negative control,and non-transfected HepG2 cells sreved as a blank ***2 cells that steadily expressed the fusion protein GPC3+α+EGFP were screened by G418,propagated,and co-cultured with lymphocytes from healthy *** proliferation was measured by the classic sulforhodamine B *** was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL),and Fas gene transcription was determined by quantitative fluorescent PCR. RESULTS:The pcDNA3.1(+)/GPC3+α+EGFP plasmid was successfully *** the experimental group,green fluorescence was observed at the cell periphery and in the cytoplasm,whereas in the negative control group,fluorescence was evenly distributed throughout the *** of the experimental group significantly decreased after 72 hours compared to the negative and blank control ***,the number of apoptotic cells was statistically different among the three groups as determined by a contingency table Chisquare test;the experimental group had the highest incidence of *** gene transcription in the experimental group was higher than in the two control groups,and an increasing trend with time in the experimental group was observed. CONCLUSION:A chimeric,membrane-anchored protein, GPC3+α+EGFP,localized

关键词： HBsAg-αepitope glypican 3 hepatocellular carcinoma hepatitis B virus protein engineering

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The targeting expression of the vascular endothelial growth factor gene in endothelial cells regulated by ***-1

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Science china(Life Sciences) 2008年第11期51卷 959-965页

作者： ZHENG XiangRong, ZHANG ShangShang, YANG YuJia, WANG Xia, ZHONG Le & YU XiaoHe department of Pediatrics, xiangya hospital, central south university, changsha 410008, china department of Pediatrics Xiangya Hospital Central South University Changsha China

The success of gene therapy depends largely on the efficacy of gene delivery vector systems that can deliver genes to target organs or cells selectively and efficiently with minimal toxicity. Here, we show that by using the ***-1 regulatory element, we were able to restrict expression of the transgene of vascular endothelial growth factor (VEGF) to endothelial cells exclusively in hypoxic conditions. Eukaryotic expression vectors such as ***-1, pcDNA3.1-VEGF+Pa, pcDNA3.1-ppET-1+ EGF+Pa, and ***-1+VEGF+Pa were constructed by using a series of nuclear molecule handling methods like PCR, enzyme digestion. The recombinant vectors were transfected into HUVEC cells and HL7702 cells by the lipofectin method. GFP expression was observed with a fluorescence microscope to validate the specificity of expression in endothelial cells under the regulation of ***-1 element. Cobalt chloride (final concentration 100 μmol/L) was added to the medium to mimic hypoxia in vitro. After transfection of vectors, the expression of VEGF mRNA was detected by RT-PCR, and the expression of VEGF was detected by Western blotting and ELISA methods under normoxia and hypoxia, respectively. The cell proliferation rate was detected by the MTT test. The ex- pression of GFP revealed that the exterior gene was transcripted effectively in endothelial cells regu- lated by the ***-1 element, while the expression of GFP was very weak in nonendothelial cells. The results of RT-PCR, Western blotting and ELISA showed that VEGF gene expression in the ***-1+VEGF+Pa group and in the pcDNA3.1-ppET-1+VEGF+Pa group was higher in hypoxia than it was in normoxia (P<0.05). The MTT test showed that the proliferation rate of HUVEC transfected with HPVA under hypoxia exceeded that of the control group. We conclude that the ***-1 element was expressed specifically in endothelial cells, and can increase the expression of VEGF in hypoxia and stimulate proliferation of

关键词： hypoxia-responsive element human preproendothelin-1 promoter vascular endothelial growth factor endothelial cells

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Modulation of TGFβ_2 and dopamine by PKC in retinal Müller cells of guinea pig myopic eye

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International Journal of Ophthalmology(English edition) 2011年第4期4卷 357-360页

作者： Jun-Feng Mao, Shuang-Zhen Liu, Wen-Juan Qin, Qian Xiang department of Ophthalmology, xiangya hospital of central south university, changsha 410008, Hunan Province, china department of Ophthalmology Xiangya Hospital of Central South University Changsha 410008 Hunan Province China.

AIM: To investigate the effect of protein kinase C (PKC) on transforming growth factor-β2 (TGFβ2) and dopamine in retinal Müller cells of guinea pig myopic eye. METHODS: Myopia was induced by translucent goggles in guinea pig, whose retinal Müller cells were cultured using the enzyme-digesting method. Retinal Müller cells were divided into 5 groups: normal control, myopia, myopia plus GF109203X, myopia plus PMA, myopia plus DMSO. PKC activities were detected by the non-radioactive methods. TGFβ2 and tyrosine hydroxylase (TH) proteins were analyzed by Western Blotting in retinal Müller cells. Dopamine was determined by the high-performance liquid chromatography- electrochemical detection in suspensions. RESULTS: After 14 days deprived, the occluded eyes became myopic with ocular axle elongating. Müller cells of guinea pigs were obtained using enzyme digestion. Compared with normal control group, the increase in PKC activity and the up-regulation in TGFβ2 expression were found in retinal Müller cells of myopic eyes, with the decrease of TH and dopamine content (P <0.05). After PKC activated by PMA, TGFβ2 and TH content were up-regulated with the increase of dopamine content (P <0.05). While the PKC activities was inhibited by GF109203X, proteins of TGFβ2 and TH were down-regulated in the myopic eyes, with the decrease of dopamine content (P <0.05). CONCLUSION: TGFβ2 and dopamine are modulated by PKC in Müller cells of the myopic eyes in guinea pig.

关键词： protein kinase C retinal Müller cell transforming growth factor-β2 dopamine myopia

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Effects of β-arrestin 2 on cytokine production of CD4+ T lymphocytes of mice with allergic asthma

Effects of β-arrestin 2 on cytokine production of CD4+ T ly...

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中华医学会呼吸病学年会——2011（第十二次全国呼吸病学学术会议）

作者： guyi wang department of Respiratory Medicine,the second xiangya hospital of central south university,changsha,Hunan 410011,china

β-arrestin 2 has been shown to participate in the pathogenesis of asthma by inducing Th2 cell migration to the *** P-arrestin 2 regulates cytokine production of CD4 T cells is still *** aim of the present study was to investigate the effect of p-arrestin 2 on the cytokine production of CD4 T lymphocytes and the mechanism involved in a mouse model for asthma.

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老年人不宜使用的药物

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中国实用内科杂志 2011年第1期31卷 17-21页

作者：蹇在金中南大学湘雅二医院老年病科长沙410011

不适当药物的使用是导致老年人发病和死亡的一个重要原因。文章介绍了国际上广泛应用的两个老年人用药指南。Beers标准介绍了48种药物在老年人中应避免使用或限量使用和20种老年人特定病症不宜使用药物的理由及危害等级。加拿大标准介绍... 详细信息

不适当药物的使用是导致老年人发病和死亡的一个重要原因。文章介绍了国际上广泛应用的两个老年人用药指南。Beers标准介绍了48种药物在老年人中应避免使用或限量使用和20种老年人特定病症不宜使用药物的理由及危害等级。加拿大标准介绍了38种药物在老年人中应避免使用,并对老年人不适当处方行为提出了替代治疗方案。推广两个指南的应用,将有利于改善老年人不适当药物应用的局面,使老年人少受药物不良反应的危害。

关键词：药物不良反应不适当药物 Beers标准加拿大标准老年人

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