1.Preoperative diagnostic value of ~(131)I-SPECT/CT fusion images in detecting the cervical and superior mediastinal metastases of DTC
Jiangong WANG ; Jun LI ; Wanning HU ; Baoming CHEN ; Ruishan ZHANG
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(04):-
OBJECTIVE To investigate the diagnostic value of 131I-SPECT/CT fusion images in detecting the metastases of differentiated thyroid carcinoma(DTC)in the regions of the head,neck and superior mediastinum.METHODS 131I-SPECT/CT fusion images was preformed for 30 patients with DTC,and the results of image were compared with the pathology after surgery.RESULTS There were 110 cases positive findings in 115 metastatic lesions with 131I-SPECT/CT fusion images examination(95.65%).5 cases of the metastatic lesions showed negative in 131I-SPECT/CT(4.35%).There was no false positive case.CONCLUSION 131I-SPECT/CT fusion images can detect and locate the metastasis and exclude the false positive results accurately.It is a good method to detect the cervical and superior mediastinal metastatic lesion in differentiated thyroid carcinoma.
2.Resveratrol regulates iNOS to inhibit atherosclerosis in C57BL/6J mice
Li QIN ; Li GUAN ; Ruishan LIN ; Qianqian XUE ; Jinwen XU ; Fuman YAN ; Xiaoying LI ; Haimei LIU
Chinese Journal of Pathophysiology 2015;(9):1601-1605
AIM:To investigate the effect of resveratrol on the lipids ( CHOL, TG, LDL-C and HDL-C) , ni-tric oxide ( NO) , peroxynitrite anion ( ONOO-) and the expression of inducible nitric oxide synthase ( iNOS) in the artery of the mice with ovariotomy ( OVX) .METHODS:The lipid levels and NO level in the serum were measured .The chan-ges of atherosclerosis were evaluated with Oil Red O staining .The expression of iNOS was measured by DAB staining and Western blot .The ONOO-production was measured by DAB staining .RESULTS:Compared with sham group , the levels of the lipids and NO production in OVX +high fat (HF) group were increased (P<0.05).Compared with OVX+HF group, the levels of the lipids and NO production in resveratrol group were decreased (P<0.05).Fourteen weeks later, the atherosclerosis model was successfully established .Compared with OVX +HF group, the iNOS expression and the ONOO-production in resveratrol group were decreased ( P<0.05 ) , while those in sham group were increased ( P <0.05).CONCLUSION:Resveratrol prevents and treats atherosclerosis by inhibiting the iNOS expression in C 57BL/6J mice.
3.Electro-acupuncture on Du meridian up-regulates the expression of Bcl-2 mRNA and its protein in early spinal cord injury of rats
Zhiying ZHANG ; Ruishan DANG ; Chuansen ZHANG ; Liang LI ; Congli REN ; Eryu CHEN
Chinese Journal of Tissue Engineering Research 2006;10(31):185-187
BACKGROUND: It is proved that acupuncture can remarkably promote recovery of nervous function after spinal cord injury (SCI). Previous studies in our groups have been proved that electro-acupuncture can inhibit apoptosis in early period of SCI, but the mechanism is unclear yet.OBJECTIVE: To study the effect of electro-acupuncture on expressions of apoptosis inhibitory gene Bcl-2 mRNA and protein with hybridization in situ and immunohistochemistry and discuss the possible mechanism of apoptosis inhibited by electro-acupuncture in early SCI.DESIGN:Opening animal study.SETTING: Department of Anatomy, the Second Military Medical University of Chinese PLA; Shanghai Acupuncture & Moxibustion and Meridian Research Center.MATERIALS:Adult male SD rats of pathogen-free aged 2-3 months were selected in this study. Bcl-2 hybridization in situ kit was provided by Wuhan Boshide Biotechnology Company Limited and Bcl-2 antibody (1:200) was provided by Santa Cruz Biotechnology Company.METHODS: The experiment was completed at the Laboratory of Anatomy of the Second Military Medical University of Chinese PLA from July 2004 to December 2005. All experimental rats were randomly divided into model group, electro-acupuncture group, methylprednisolone group and sham operation group. T10 spinal cord was injuried by the modified Allen's method and treated with electro-acupuncture immediately, and then the expressions of Bcl-2 mRNA and protein were evaluated with hybridization in situ and immunohistochemistry combined with image quantitative analysis.MAIN OUTCOME MEASURES: ① Expressions of Bcl-2 mRNA and protein in early SCI; ② effect of electro-acupuncture on expressions of Bcl-2 mRNA and protein.RESULTS: The rats were supplied when they died during the experiment,and all 42 rats were involved in the final analysis. ① Moderate expression of Bcl-2 mRNA and protein was observed in the sham operation group.Expression of Bcl-2 mRNA was increased in model group at 6 hours after SCI, but expression ofBcl-2 protein was not changed. At 24 hours after SCI, both expressions of Bcl-2 mRNA and Bcl-2 protein were increased.Expression of Bcl-2 mRNA and protein was higher in electro-acupuncture group than that in mo del group (P < 0.05), and there was no significant difference from that in methylprednisolone group. ② Amount of positive Bcl-2 mRNA cells was increased in electro-acupuncture group and methylprednisolone group at 6 hours after treatment, and gray value was decreased.There was significant and remarkably significant difference from those in sham operation group and model group, respectively (P < 0.05-0.01). After 24-hour treatment, amount of positive Bcl-2 mRNA cells was increased in electro-acupuncture group and gray value was decreased.There was significant and remarkably significant difference from those in sham operation group and model group, respectively (P < 0.05-0.01). ③ After 24-hour treatment, amount of immunohistochenistry positive Bcl-2 cells was increased in electro-acupuncture group and gray value was decreased. There was significant and remarkably significant difference from those in sham operation group and model group, respectively (P < 0.05-0.01).CONCLUSION: Electro-acupuncture can up-regulate the expressions of Bcl-2 mRNA and protein so as to inhibit apoptosis in early SCI.
4.Research progress in aromatic prenyltransferases originated from microorganisms.
Xiao LIU ; Ridao CHEN ; Dan XIE ; Jianhua LI ; Ruishan WANG ; Jungui DAI
Acta Pharmaceutica Sinica 2013;48(2):161-9
The prenylation of aromatic compounds plays an important role in the natural product research because it not only gives rise to an astounding diversity of primary and secondary metabolites in plants, fungi and bacteria but also enhances the bioactivities and bioavailabilities of these compounds. However, further investigation of prenylated aromatic compounds is frequently hindered due to their low content in nature and difficulties in chemical synthesis. Cloning aromatic prenyltransferase genes followed by heterologous expression would be attractive tools for the chemoenzymatic synthesis of bioactive molecules. This review summarizes the classifications, structural investigations, enzymatic catalysis and other progress in aromatic prenyltransferases originated from microorganisms.
5.Identification of neural tissue-committed stem cells, a new cellular group, in bone marrow
Zhiying ZHANG ; Congli REN ; Chuansen ZHANG ; Liang LI ; Ruishan DANG ; Zhengdong KONG
Chinese Journal of Tissue Engineering Research 2008;12(8):1577-1580
BACKGROUND: It has been widely accepted that both bone marrow-derived mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs) have the capacity to differentiate into neural lineages. Some scholars believe that in addition to HSCs and MSCs, bone marrow (BM) also harbors a highly mobile population of CXCR4+ tissue committed stem cells (TCSCs), including skeletal muscles, heart, liver, and neural tissue. OBJECTIVE: To make sure that neural tissue-committed stem cells (NTCSCs) reside in the bone marrow, and to establish a purification and culture method for bone marrow-derived NTCSCs.DESIGN: Opening animal study.SETTING: Department of Anatomy, the Second Military Medical University of Chinese PLA. MATERIALS: Adult Sprague-Dawley (SD) rats (pathogen-free) were provided by the Animal Center of the Second Military Medical University of Chinese PLA. Dulbecco's modified eagle's medium (DMEM)/F12, B27, N2 and epidermal growth factor (EGF, Invitrogen Company), basic fibroblast growth factor (bFGF, CytoLab Ltd), rabbit anti-rat Nestin,CXCR4, β-Tublin Ⅲ, glial fibrillary acidic protein (GFAP, Santa Cruz Company), mouse anti-rat microtubule associated protein 2ab (MAP2ab) (Clone11-5B), cyclic nucleotide 3'phosphohydrolase (CNPase, Clone AP20, NeoMarkers Company), fluorescent(fluorescein isothiocyanate, Cy3) marker reagents (Wuhan Boster Bioengineering Co., Ltd), nuclear fluorescent dyes 4,6-diamidino-2-phenylindole(DAPI)(Sigma), immunohistochemistry reagents (Vector Laboratories Company) , and NycoPrepTM separation liquid (1.077A, Axis-Shield Company) were used in this study.METHODS: This study was performed in the Department of Anatomy, the Second Military Medical University of Chinese PLA from January 2004 to December 2006. Bone marrow was harvested from bilateral femurs and tibias of 2-3 weeks SD rats. Mononuclear cell layer was isolated by NycoPrepTM separation liquid and suspended in DMEM/F12(1:1)serum-free medium supplemented with 2% B27,1% N2, 20 μg/L bFGF, 20μg/L EGF, 1×105 U/L penicillin and 100 mg/L streptomycin. NTCSCs were isolated and propagated by suspensive growing from adherent cells in bone marrow in DMEM/F12 free-serum medium. MAIN OUTCOME MEASURES: NTCSCs were identified by immunocytochemistry for CXCR4, a marker of TCSCs and nestin, a marker of neural stem cells, and neural lineages marker protein after differentiation of cellular spheres. RESULTS: The NTCSCs spheres expressed nestin, a neural stem cell marker as well as CXCR4, a marker of TCSCs. The NTCSCs' spheres were naturally differentiated in DMEM medium with 15% fetal bovine serum. The differentiated cells expressed β-Tublin Ⅲ, MAP2ab, CNPase and GFAP, markers of neural lineages. CONCLUSION: NTCSCs reside in bone marrow and naturally differentiate into neural lineages in vitro.
6.Polymorphism analysis on chemokine SDF-1α transcript being the ligand for HIV-1 coreceptor CXCR4
Xiuying ZHAO ; Hao WU ; Ruishan LI ; Jiaqing HUANG ; Chiwai CHAN ; Bojian ZHENG
Chinese Journal of Laboratory Medicine 2008;31(5):523-527
Objective To determine the polymorphism in CXC chemokine SDF-1α transcript and its effects on HIV infection.Methods Three groups of study subjects lived in Hang Kong were recruited:278 HIV-heahhy donors of Chinese origin.49 HIV+Caucasians and 13 Chinese with high risk behavior to HIV but kept uninfected.Genomic DNA and RNA were extracted from eripheral blood mononucleal cell. The PCR and RT-PCR reaction were set up accordingly.Sequence of the SDF-1α promoter,the open reading frame(ORF)and the 3'untranslate region(3'UTR)were analyzed.Two steps PCR reaction using two reverseprimers with mismatched nucleic acid were employed to screen the frequency of a novel mutation. Results equencing analysis from 100 subjects indicated that non mutation happened in tlle promoter and ORF of SDF-1α.A novel mutation Was detected from 3'UTR of SDF-1α.It is a "GA" insertion in "G" rich region near the stop code of SDF-1α.The mutation Was named as SDF-1-3’GA+and submitted to GenBank (AY874118).The mutation happened in three roups.with allele frequency of 15.1% in the healthy Chinese.of 30.7% in the high risk Chinese group.Conclusions our results confirm that SDF-1 genes arerelatively conserved.None noteworthy mutation is identified in the promoter and ORF regions of SDF-1α However.a novel mutation is identified from the 3'UTR of SDF-1α. It would be worthwhile to etermine effect of the novel mutation on HIV infection.
7.Expression of prokaryotic ubiquitin-like protein (Pup)-proteasome system in drug-resistant Mycobacterium tuberculosis
Li HE ; Ying LEI ; Fang WU ; Le ZHANG ; Jiangdong WU ; Xudong CAO ; Bin ZHU ; Bo WU ; Ruishan LI ; Yuqing ZHANG ; Zhao WANG ; Wanjiang ZHANG
Chinese Journal of Immunology 2015;(6):741-747,752
Objective:To study the different gene expressions of Pup-proteasome system between the original drug resistance Mtb and the cultured Mtb( INH-Mtb,RFP-Mtb,SM-Mtb,EB-Mtb,MDR) which were at the different concentrations of the Mtb drug,and to explore whether the different Pup-proteasome system gene expression was relevent to the clinical isolates of Mtb-resistant which was widely spreaded in Xinjiang region. Methods:Culturing INH-Mtb,RFP-Mtb,SM-Mtb,EB-Mtb,MDR strains at Original drug resistance Medium,low concentrations of drug Medium and high concentrations of drug Medium to the logarithmic phase,and extract total RNA from each group of Mtb. Using SYBR GreenⅠreal-time PCR to detect the Pup-proteasome system expression level in different concen-trations of drug Mtb in each group of Mtb. Results: Compared with the original state of drug-resistant strains,Pup gene in INH-Mtb, RFP-Mtb,SM-Mtb and MDR strains group were down-regulated 0. 74,0. 23,0. 28,0. 57 times;Dop gene were up-regulated 1. 33,1. 63, 1. 14,2. 88 times;PafA gene were up-regulated 1. 69,1. 30,1. 58,1. 32 times;Mpa gene were up-regulated 3. 05,1. 79,1. 31,2. 27 times in low concentrations of Anti-Mtb drugs condition, the difference was statistically significant ( P<0. 05 ) . Compared with the original state of drug-resistant strains,Pup gene in INH-Mtb,RFP-Mtb,SM-Mtb,MDR strains group were down-regulate 0. 58,0. 37, 0. 43,0. 78 times;Dop gene were up-regulated 2. 62,2. 49,1. 69,2. 95 times;PafA gene were up-regulated 2. 16,1. 48,2. 02,2. 21 times;Mpa gene were up-regulated 1. 63,3. 22,1. 13,3. 94 times in the high concentrations of Anti-Mtb drugs condition,the difference was statistically significant(P<0. 05). Conclusion:Through the different concentrations of antibiotic selection pressure,these groups of Mtb strains expressions in the Pup-proteasome system of Pup gene,Dop gene,Mpa gene and PafA gene were different,The results reveal that Pup-proteasome system is associated with the drug resistance in Mtb which was spreaded in Xinjiang region.
8.Correlation study between prokaryotic ubiquitin-like protein (Pup)-proteasome system of Mycobacterium tuberculosis and persistence of Mycobacterium tuber-culosis
Bin ZHU ; Ying LEI ; Fang WU ; Hui ZHANG ; Chunjun ZHANG ; Le ZHANG ; Jiangdong WU ; Xudong CAO ; Bo WU ; Li HE ; Yuqing ZHANG ; Ruishan LI ; Zhao WANG ; Wanjiang ZHANG
Chinese Journal of Immunology 2015;(4):447-452
Objective:To explore the regulation of prokaryotic ubiquitin-like protein ( Pup )-proteasome system on the persistence of Mycobacterium tuberculosis by inducing Mycobacterium tuberculosis to being persistence state under hypoxia conditions and then analyzing the difference on the expression levels of Pup,Dop,PafA and Mpa gene at various time and different conditions. Methods:The total mRNA of the international standard virulent strains of Mycobacterium tuberculosis(H37Rv),which were cultured under hypoxia and aerobic conditions, were extracted from each group at various time and purity of the mRNA were identified.The expression of Pup,Dop,PafA and Mpa genes of M.tuberculosis strains in each group were quantified by SYBR Green I qRT-PCR,which aimed at finding the difference among the expression of Pup,Dop,PafA and Mpa genes at various time and different conditions.Results:The expression levels of Pup, Dop, PafA and Mpa genes in Mycobacterium tuberculosis under hypoxia conditions were measured at various times.The expression levels of Pup,Dop,PafA and Mpa genes:compared with the 0 d,the expression of the Pup gene was up-regulated 1.66,2.43 and 2.76-fold at 4 d,7 d,10 d respectively(P<0.05);the expression of the Dop gene was up-regulated 1.38, 1.91,2.54 and 3.28-fold at 2 d,4 d,7 d,10 d respectively(P<0.05);the expression of the PafA gene was up-regulated 1.22,1.75, 2.37 and 2.67-fold at 2 d,4 d,7 d,10 d respectively( P<0.05);the expression of the Mpa gene was up-regulated 1.66,2.21 and 2.63-fold at 4 d,7 d,10 d respectively(P<0.05).Take the aerobic conditions as control,under hypoxic conditions with the same culture time,the expression of the Pup gene was up-regulated 1.85,2.81 and 2.93-fold in 4 d,7 d,10 d respectively(P<0.05);the ex-pression of the Dop gene was up-regulated 1.20,1.76,2.01 and 3.01-fold in 2 d,4 d,7 d,10 d,respectively( P<0.05);the expression of the PafA gene was up-regulated 1.22,1.57,2.29 and 2.29-fold in 2 d,4 d,7 d,10 d,respectively(P<0.05);the expression of the Mpa gene was up-regulated 1.16,1.58,2.16 and 2.69-fold in 2 d,4 d,7 d,10 d,respectively(P<0.05).Conclusion:Under hypoxic conditions,there were significant differences on the expressions of Pup, Dop, PafA and Mpa genes at various times;what′s more, significant differences on the expressions of Pup,Dop,PafA and Mpa genes exist between hypoxic and aerobic conditions at the same time,Prokaryotic ubiquitin-like protein( Pup)-proteasome system plays a regulatory role on M.tuberculosis′s persistence.
9.Correlation study between prokaryotic ubiquitin-like protein (Pup)-proteasome and drug resistance of clinical isolates of Mycobacterium tuberculosis
Li HE ; Ying LEI ; Fang WU ; Le ZHANG ; Jiangdong WU ; Xudong CAO ; Bin ZHU ; Bo WU ; Ruishan LI ; Yuqing ZHANG ; Zhao WANG ; Rui ZHUANG ; Wenjuan LI ; Chen LIANG ; Chao FAN ; Wanjiang ZHANG
Chinese Journal of Immunology 2014;(11):1441-1447,1451
Objective:To study the correlation different drug-resistant mycobacterium tuberculosis clinical isolates of prokaryotic ubiquitin-like protein ( Pup )-proteasome of Pup, Dop, PafA, Mpa gene expression level and mycobacterium tuberculosis Pup-proteasome system with Xinjiang region widely popular drug-resistant mycobacterium tuberculosis in clinical isolates resistance.Methods:Total RNA of Mtb was extracted from cultured Mtb during the logarithmic phase in drug-susceptible strains in Xinjiang region,the clinical strains drug sensitive to INH,RFP,SM and EB respectively,and multidrug-resistant(MDR) strains.And then the purity of total RNA was identified.The expressing of Pup-proteasome relevant gene( Pup,Dop,Mpa,PafA) were quantified using SYBR GreenⅠqRT-PCR which aimed at finding the correlation between Mtb Pup-proteasome system and drug resistance of Mtb clinical isolates widespread in Xingjiang region by analyzing the expression of Pup, Dop, PafA, Mpa gene among different isolates.Results:Compared with the drug-senstive clinical isolates, mRNA expression level of Pup, Mpa gene was down-regulated in resistant M.tuberculosis clinical isolates INH ( INH-MTB) ,RFP ( RFP-MTB) ,SM ( SM-MTB) and EB ( EB-MTB) ,mRNA expression levels of genes in Dop and PafA was higher in resistant M.tuberculosis clinical isolates,the difference was statistically significant(P<0.05).Compared with MDR strain, the expression of Pup, Dop, Mpa gene were up-regulated different in the resistant M.tuberculosis clinical isolates isolates ,the expression of PafA gene was down-regulated different,the difference was statistically significant( P<0.05).Conclusion:The differentially expressed gene of Pup、Dop、PafA、Mpa gene in sensitive strains,INH-MTB,RFP-MTB,SM-MTB,EB-MTB and MDR strains.The Mycobacterium tuberculosis Pup-proteasome system.Therefore,the Pup-proteasome system have association with the drug resistance of Mtb strains widespread in Xinjiang region.
10.Correlation study between Mycobacterium tuberculosis PhoPR two-component system and pathogenicity of different virulent Mycobacterium tuberculosis
Bo WU ; Fang WU ; Le ZHANG ; Jiangdong WU ; Hui ZHANG ; Chunjun ZHANG ; Xudong CAO ; Bin ZHU ; Ruishan LI ; Li HE ; Zhao WANG ; Yuqing ZHANG ; Rui ZHUANG ; Wenjuan LI ; Chen LIANG ; Chao FAN ; Wanjiang ZHANG
Chinese Journal of Immunology 2014;(10):1320-1324,1329
Objective:To explore the correlation between Mycobacterium tuberculosis PhoPR two-component system and the pathogenicity of different virulent MTB by analysing the expression levels difference of PhoP gene and PhoR gene in BCG ,H37Ra, H37Rv and XJ-MTB respectively.Methods:Total RNA extracted from four different virulent MTB strains and the integrity of total RNA were identified by using agarose gel electrophoresis.The expression of PhoP gene and PhoR gene were quantified by using SYBR Green I FQ-PCR.The expression levels difference of these genes were compared in different virulent MTB strains .Results: The relative expression levels of PhoP gene in between four different virulent MTB strains from high to low were XJ -MTB(9.05),H37Rv(1.00), H37Ra(0.25),BCG(0.08) respectively ,and the expression levels difference of PhoP gene were statistically significant in different virulent MTB strains ( P<0.05 );the relative expression levels of PhoR gene in four different virulent MTB strains from high to low were XJ-MTB(5.72),H37Rv(1.00),H37Ra(0.18),BCG(0.07) respectively,and the expression levels difference of PhoR gene were sta-tistically significant in different virulent MTB strains ( P<0.05 ).The expression levels of PhoP gene and PhoR gene at XJ-MTB were statistically significant difference compared with BCG ,H37Rv,H37Ra respectively (P<0.05);the expression levels of PhoP gene and PhoR gene at H37Rv were statistically significant difference compared with BCG ,H37Ra respectively (P<0.05);the expression levels of PhoP and PhoR gene at BCG were not statistically significant difference compared with H 37Ra (P>0.05).Conclusion:Significant expression levels difference of PhoP gene and PhoR gene are confirmed in different virulent MTB strains .Therefore,the Mycobacterium tuberculosis PhoPR two-component system is correlated with the pathogenicity in different virulent MTB strains.