2.Mutations in the rpsE gene and spectinomycin resistance in Neisseria gonorrhoeae
Wen ZHU ; Faxing JIANG ; Xiaohong SU ; Wenjing LE ; Na WANG
Chinese Journal of Dermatology 2014;47(9):611-614
Objective To evaluate the relationship between spectinomycin resistance in Neisseria gonorrhoeae and mutations in the rpsE gene.Methods Genomic DNA was extracted from 4 clinical isolates of Neisseria gonorrhoeae with different levels of spectinomycin resistance.Then,PCR was performed to amplify the entire rpsE gene and the spectinomycin resistance-determining region (SRDR) in the 16S rRNA gene followed by direct sequencing.Two spectinomycin-sensitive Neisseria gonorrhoeae strains were transformed with the genomic DNA containing the mutant rpsE gene.Subsequently,the susceptibility of the transformants to spectinomycin was determined,and PCR was performed to amplify the rpsE and 16S rRNA genes in the transformants followed by sequencing.Results All the 4 spectinomycin-resistant Neisseria gonorrhoeae strains harbored an A70C transversion in the rpsE gene,but no abnormality in the SRDR of the 16S rRNA gene.No mutations were detected in the spectinomycin-sensitive Neisseria gonorrhoeae strains.The A70C transversion in the rpsE gene was also detected in the two Neisseria gonorrhoeae transformants with spectinomycin resistance.Conclusion The A70C point mutation within the rpsE gene is associated with spectinomycin resistance in Neisseria gonorrhoeae.
3.Recombinant mutant human tumor necrosis factor enhances anti-angiogenesis effect of cisplatin on Lewis lung carcinoma in mice
Su-Ju WEI ; Jiang-Lian YOU ; Wen-Feng HAN ; Yi-Bing LIU ;
Chinese Journal of Cancer Biotherapy 1995;0(03):-
Objective: To observe the role of rmh-TNF in enhancing the anti-angiogenesis effect of cisplatin on Lewis lung carcinoma in the mice.Methods: Lewis lung carcinoma model was established in C57BL/6 mice.Sixty model mice were randomly divided into 4 groups: control group,rmh-TNF group(1500000 U/kg),cisplatin group(6.15 mg/kg), and rmh-TNF plus cisplatin group.Twelve days after implantation of cancer cells,different drugs were injected intra- tumorallv for 3d.The expression of hypoxia inducible factor-1?(HIF-1?)gene in the tumor was identified by RT-PCR. Immunohistochemistry(IHC)image analysis was performed to determine the vascular endothelial growth factor(VEGF) and kinase domain region receptor(KDR)expression and the microvessel density(MVD).Expression of matrix metallo- proteinase-2(MMP-2)was detected by flow cytometry.Results: The MVD values in the control group,the rmh-TNF group,the DDP group and the combination group were(24.76?1.28),(18.95?1.22),(19.53?1.15),(10.43?1.05),respectively,with those of the rmh-TNF and DDP groups significantly lower than that of the control group and higher than that of the combination group(all P
4.Effect of Bioimprinting by Lauric Acid on Esterification Activity of Lipase
Xiong-Wen CAO ; Lei SU ; Qi-Shen CHEN ; Tao LIU ; Yun-Jun YAN ; Jiang-Ke YANG ;
China Biotechnology 2006;0(07):-
Bioimprinting is a new developed technique to improve the characteristics of enzymes.Bioimprinting by lauric acid was conducted to improve the esterification activity of lipase PS in sol-gel immobilization process with methyltrimethoxysila(MTMS) and tetramethoxysila(TMOS) as the precursors.Results generated by checking the esterification activity and scanning electron microscope showed that bioimprinting can enhance the specific activity and thermal stability of lipase PS.The bioimprinting system was optimized by orthogonal experiment,and the optimal condition for lipase bioimprinting is water/silane molar ration(R) 12,polyethylene glycol(PEG) 120?l,and lauric acid 0.15 mmol.Compared with the free enzyme and the non-imprinted enzymes,the specific activity of imprinted enzymes has been improved 44.3 fold and 2.4 fold,respectively.Imprinted lipase show better thermal stability,and the relative activity is 58% after incubated in 80 ℃ for 0.5 h,while no activity was detected for the free enzyme.
5.The therapeutic effects of electroacupuncture on post-stroke depression after lacunar infarct
Hong JIANG ; Xin-Yi WEN ; Ting-Hui SHI ; Su-Ming ZHANG ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(12):-
Objective To investigate the therapeutic effects of electroacupuncture (EA) on post stroke depres- sion (PSD) after lacunar infarction (LI).Methods Seventy-five PSD patients with lacular infarction were recruited and randomly divided into a control group and an EA group.All patients were treated with Fluoxetine,in addition to EA treat- ment in the EA group.Then all patients were evaluated using the Hamilton Depression Scale (HAMD),the Chinese stroke scale (CSS) and the Barthel Index (BI).The evaluations were carried out at 0,14 and 28 days.Results Depression symptoms (DSs) in the EA group were improved by day 14 of the treatment,and their HAMD scores had decreased.DSs in both groups had improved significantly by day 28,and the HAMD scores in the EA group were then significantly lower than those in the control group.Improvements in neurological impairment and in the activities of daily living were observed earlier in the EA group.Conclusion The combination of EA and Fluoxetine is helpful for PSD with complex patho- genesis and clinical symptoms.Therapeutic effects are enhanced and side effects are reduced.
6.Effect of bcl-xi overexpression in transgene mice with cerebral infarction and study of cytochrome Cexpression and caspase-3 expression
Fu-Rong WANG ; Yong-Sheng JIANG ; Yan LIU ; Wen-Wu XIAO ; Su-Ming ZHANG ;
Chinese Journal of Emergency Medicine 2006;0(09):-
0.05).At different time points after ischemia-reperfusion,the expression of cytochrome C and activation of caspase-3 were lower in the transgen mice than that in the wild type rats.Conclusions Under standard condition,overexpression of bcl-xl could significantly reduce the infarct area and improve neurological function in transgene mice than those in the wild type rats.The effect of overexpression of bcl-xl might be realized through inhibiting the apoptosis of neuron,and the mechanism might be that the overexpression of bcl-xl inhibit the release of cytochrome C and the activation of caspase-3.
7.Expression and significance of Oct4 B1 in colorectal cancer stem cells
Jiaping CHENG ; Jiang LI ; Xian SU ; Yilin CHEN ; Qingliang ZENG ; Kunming WEN
Chinese Journal of Immunology 2016;32(6):794-797
Objective:To investigate the expression and its possible role of Oct4B1 subtype of Embryonic stem cell transcription factor Oct4 in colorectal cancer stem cells. Methods: 3D microspheres were cultured by suspension culture to human colorectal cancer cell line SW480 cells. The 3D microspheres and SW480 cells were used as the research objects. Whether 3D microspheres were enriched cancer stem cells,we used the methods of cell differentiation experiments,soft agar cloning experiments,and the expression levels of cancer stem cells markers CD133,CD44 detected by flow cytometry. The expression levels of Oct4B1 mRNA were detected by RT-qPCR. Results:3D microspheres could differentiate into normal cancer cells. Compared with the parental SW480 cells,in vitro colony formation was significantly enhanced(P<0. 01),the percentage of positive cells of CD133 and CD44 were significantly increased ( P < 0. 01 ), the expression levels of Oct4B1 mRNA were obviously higher ( P < 0. 01 ) in 3D microspheres. Conclusion: Oct4B1 subtype of Embryonic stem cell transcription factor Oct4 in 3D microspheres enriched human colorectal cancer stem cells,which may be involved in the regulation of colorectal cancer stem cells.
8.A Study on Quality of the Precise Powder Decoction Pieces of Medicinal Flowers Lonicerae japonicae Flos
Zhiyao REN ; Wen XU ; Jing ZHANG ; He SU ; Linlin DONG ; Jiang XU ; Xiaohui QIU ; Zhihai HUANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(1):89-94
This study aimed at evaluating the quality of the precise powder decoction pieces (PPDP) of L.japonicae Flos (LJF) compared with the traditional commercial slices with chemical fingerprint methods and DNA molecular identification technology.Different specifications of PPDP were prepared,their dry extract contents were in contrast with that of commercial slices.The three batches of commercial slices were collected,and the content uniformity,fingerprint and similarity evaluation before and after the mixing and pulverization were studied by HPLC-DAD and DNA sequence alignment.As a result,the paste rate of PPDP was slightly higher than that of the traditional commercial slices.The dissolution of chlorogenic acid of PPDP was higher than that of the traditional commercial slices.RSD of inter-assay dissolutions of chlorogenic acid of commercial slices was 11.93%,which was reduced to 8.29% after mixing and preparing into PPDP.The fingerprint showed that the slimilarity of the fringerprint of the mixed and powdered LJF was elevated with 7 common peaks.All the common peaks were increased at different levels.In conclusion,compared with traditional commercial slices of LJF,PPDP apparently improved the dissolution rate and the quality uniformity,indicating that the boiled powder of CRP obviously presented vantages in clinic.
9.Changes and function of Bax and Fas in HL-60 cell apoptosis induced by two fungus polysaccharides
Henglan ZHANG ; Peie WEN ; Xia REN ; Xiaobai SUN ; Enyu SU ; Guosheng JIANG
Journal of International Oncology 2009;36(12):946-948
Objective To detect the proliferation inhibition and apoptosis of HL-60 cell induced by APS-1 and APS-2 isolated from Polyporus sp. M05 and to investigate its mechanism. Methods The proliferation inhibition was detected by living cells count method,and chosed proper concentration.Flow cytometry with propidium iodide staining was used to detect cell apoptosis. Semi-quantitative RT-PCR was used to detect the expression of apoptosis related gene. Results APS-1 and APS-2 could significantly inhibit the proliferation of HL-60 cells on a time and dose dependent manner. Apoptosis ratio increased to 47. 9% and 26. 8% after HL-60 cells were exposed to APS-1 and APS-2 respectively for 48 h,and the differences had statistical significance (P <0.01). After being induced by APS-1,mRNA of Bax,Fas,Caspase-3 was upregulated. And after being induced by APS-2,mRNA of Bax,Caspase-3 was upregulated,while Fas mRNA did not change. Conclusion APS-1 and APS-2 can inhibit the proliferation and induce apoptosis of HL-60 cells. Mechanism of HL-60 cell apoptosis induced by APS-1 is related to both mitochondrial pathway and Fas signaling pathway,while apoptosis induced by APS-2 is only related to mitochondrial pathway.
10.Effect of TGF-β1/SMAD signaling pathway on K562 cells growth inhibition caused by HMBA
Enyu SU ; Peie WEN ; Xia REN ; Xiaobai SUN ; Henglan ZHANG ; Tianhua TANG ; Haiquan REN ; Guosheng JIANG
Journal of International Oncology 2010;37(4):312-315
Objective To investigate the effect of TGF-β1/SMAD signaling pathway on K562 cells growth inhibition caused by HMBA. Methods After establishing the in vitro differentiation model with HMBA on K562 cells, the MTT assay was used to detect the proliferation of K562 cells, the cell cycle profile was detected by flow cytometry, and the mRNA expression of TGF-β1, SMAD3, SMAD4 and EVI1 was measured by RT-PCR assay. Results HMBA could inhibit the proliferation and promote the differentiation of K562 cells obviously, which was time and concentration-dependent, and the 72 h corresponding IC50, was about 2 mmol/L. Within 72 h, flow cytometry assay indicated that the ration of G0-G1 phase cells was up-regulated, and the results of RT-PCR showed that relative mRNA expression of TGF-β1, SMAD3 and SMAD4 at mRNA level was increased gradually while that of EVI1 was decreased gradually. Conclusion HMBA can inhibit K562 cells proliferation through TGF-β1/SMAD signaling pathway.