1.Expression, purification and effect of different immunization approaches of Tir from EHEC O157:H7
Ling WANG ; Liping HU ; Beiguo LONG ; Yong ZHOU ; Jun LUO ; Guangming YUANG ; Hongying FAN
Chinese Journal of Microbiology and Immunology 2011;31(4):335-340
Objective To clone and express translocation intimin receptor(Tir)of enterohemorrhagic Escherichia coli(EHEC)O157:H7,and to analyze the effect of different routes on the induction of immunity to the recombinant protein.Methods The tir eucoding genes were amplified from EHEC O157:H7 strain guangzhou 246 genome,and genes were cloned into the vector pET-30a(+).The pET-30a(+)tir recombinant was transformed into E.coli BL21.and expression was induced bv IPTG.The expressed product was analyzed by SDS-PAGE and purified by Ni-IDA affinity chromatography.The immunized mice sera and fecal against the recombinant protein was detected.Resuits The length of the tir is 1677 bp,with the initiation codon ATG and the termination codon TAA.Double enzyme digestion and DNA sequencing confirmed that the recombinant expression plasmid pET-30a(+)-tir was constructed.The recombinant protein was expressed in Escherichia coli expression system,and was purified by Ni-IDA affinity chromatography.The mice were able to produce a high serum IgG antibody titer after both subeutaneous and intranasal immunizations.Meanwhile,the intranasal immunization induced serum and fecal IgA antibody titer was significantly higher than that of the subcutaneous immunization group.Conclusion Tir molecule is potential vaccine candidate for preventing EHEC disease.
2.Analysis of hysteroscopy results of 539 patients accept in vitro fertilization-embryo transfer
Qian PENG ; Yuyan LI ; Ling LONG ; Min LI ; Qinghua QU ; Hong LUO
Chongqing Medicine 2015;(16):2217-2218,2221
Objective To evaluate uterine cavity of the patients who accept the treatment in vitro fertilization‐embryo trans‐fer(IVF‐ET)and determine the clinical value of hysteroscopy for the patients .Methods Between January 2012 and May 2014 ,539 cases who accepted the IVF‐ET and hysteroscopy ,were divided into two grops ,group A(258 patients examinated before IVF‐ET cycle) and group B(281 patients after the IVF‐ET failure) ,and retrospective analysis was performed for their uterine cavity and cervix canal data .Results The total incidence of abnormal hysteroscopic findings was 49 .54% ,and group B was significantly high‐er than that in group A(54 .10% vs .44 .57% ,χ2 =4 .5 ;P=0 .043) .The percent of abnormal cervix canal was 18 .74% ,and there wasn′t significant difference between the two groups .Conclusion The incidence of abnormal hysteroscopic findings in IVF‐ET is higher than that in normal patients ,so hysteroscopy has an important clinical value for diagnosis and evalution ,especially for pa‐tients with failure of IVF‐ET .
3.A Study on Anti-EV71 Effects of Ganlu Xiaodudan in Vitro
Bichen AI ; Youshun HE ; Guorong ZHAO ; Yirong HE ; Ling LONG ; Can LI ; Chengyu LUO
Chinese Journal of Information on Traditional Chinese Medicine 2014;(11):62-65
Objective To explore anti-EV71 effects of Ganlu Xiaodudan in vitro. Methods Ribavirin was taken as control drug, with the help of cell culture to observe anti-EV71 inhibition rate of Ganlu Xiaodudan in inhibiting-virus-directly experiment, therapeutic-inhibiting-virus experiment, preventive-inhibiting-virus experiment and preventive-therapeutic-inhibiting-virus experiment. Results In inhibiting-virus-directly experiment, therapeutic-inhibiting-virus experiment and preventive-therapeutic-inhibiting-virus experiment, virus inhibition rate of Ganlu Xiaodudan was higher than ribavirin. In preventive-inhibiting-virus experiment, virus inhibition rates of Ganlu Xiaodudan and ribavirin both were almost zero. Conclusion Ganlu Xiaodudan has better antiviral effects on EV71 than ribavirin, and it can affect more than one link of multiplication of EV71.
4.The role of STAT3 in inflammatory bowel disease and colitis-associated cancer and research progress of the related drugs
Xiao-fan CHENG ; Hu-tai-long ZHU ; Ling LIU ; Jing LUO ; Zhi-jie SUN ; De-li DONG
Acta Pharmaceutica Sinica 2022;57(8):2253-2261
Signal transducer and activator of transcription (STAT) 3 is a critical transcription factor for cell proliferation and survival. It is activated within cells by many cytokines to mediate immune and inflammatory responses to injury. Inflammatory bowel disease (IBD), represented by Crohn′s disease (CD) and ulcerative colitis (UC), is a chronic inflammatory disease of the intestinal tract. STAT3 has been shown to be abnormally activated in IBD colon tissues by many pro-inflammatory cytokines, leading to disruption of the intestinal mucosal barrier and excessive innate immune and Th17 responses. The persistent chronic inflammation eventually leads to intestinal fibrosis and stenosis. In addition to immune responses, STAT3 is also involved in intestinal fibrosis in IBD by promoting the transcription of fibrosis-related genes. Colitis-associated cancer (CAC) is a particularly aggressive subtype of colorectal cancer and is associated with chronic inflammation-induced IBD. STAT3 has also been associated with CAC initiation and development. STAT3 is overactivated in tumors, which leads to suppression of the anti-tumor activity of immune cells and promotion of cancer cell proliferation, tumor angiogenesis, invasion, and migration. In the present article, we summarize the role of STAT3 in IBD and CAC and the research progress of the related drugs developed for UC and CAC treatment.
6.Murine model of busulfan-induced spermatogenesis regeneration: a quantitative evaluation.
Xiao-min LUO ; Ci ZHANG ; Si-xing YANG ; Ling-long WANG
National Journal of Andrology 2010;16(5):395-399
OBJECTIVETo quantitatively evaluate the murine model of spermatogenesis regeneration induced by two-dose busulfan injection.
METHODSFifty-four male mice were randomly divided into a control and two model groups of equal number, the former treated by two-dose intraperitoneal injection of 50% DMSO solution at 10 ml/kg, and the latter by that of busulfan at 10 mg/kg and 15 mg/kg respectively to establish spermatogenesis regeneration models, both at the interval of 24 days between the two doses. Spermatogenesis in seminiferous epithelia was evaluated by Johnsen score, and the expressions of GATA-4 and GDNF mRNA in Sertoli cells were detected by real time quantitative PCR at 3, 4 and 8 weeks after the treatment.
RESULTSJohnsen score kept stable in the control group at all stages (P > 0.05), but higher than in the model groups at 3 and 4 weeks (P < 0.01). It was lower in the 15 mg/kg than in the 10 mg/kg model group at 4 and 8 weeks (P < 0.01) , and than in the control group at 8 weeks (P < 0.05), but had no significant difference between the 10 mg/kg and the control groups (P > 0.05). Nor did the expression of GATA-4 mRNA in Sertoli cells show any significant difference among the three groups at different stages after the treatment (P > 0.05), and that of GDNF mRNA at different stages in the control group (P > 0.05). Compared with the controls, the level of GDNF mRNA in Sertoli cells was significantly higher at 3 weeks but lower at 4 weeks in the model groups (P < 0.01), and lower in the 15 mg/kg group (P < 0.01) and comparable in the 10 mg/kg group at 8 weeks (P > 0.05); and it was lower in the 15 mg/kg than in the 10 mg/kg group at all stages (P < 0.01).
CONCLUSIONTwo-dose intraperitoneal injection of 10 mg/kg busulfan at the interval of 24 days is an optimal option for the establishment of a murine model of spermatogenesis regeneration. Higher dose of busulfan may induce deficient expression of GDNF in Sertoli cells and result in incomplete restoration of spermatogenesis.
Animals ; Busulfan ; adverse effects ; Glial Cell Line-Derived Neurotrophic Factor ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Models, Animal ; RNA, Messenger ; Regeneration ; drug effects ; Sertoli Cells ; drug effects ; Spermatogenesis ; drug effects ; Spermatozoa ; physiology ; Testis ; drug effects ; physiology
7.Study on Attitudes towards Deprescribing among Elderly Patients with Chronic Diseases in Several Communities of Chongqing
Sha ZHANG ; Long MENG ; Jiadan YANG ; Ling LUO ; Feng QIU
China Pharmacy 2018;29(10):1408-1411
OBJECTIVE:To provide reference for promoting smoothly implementation of deprescribing. METHODS:A total of 335 elderly patients with chronic disease were randomly selected from 4 communities and 9 village clinic of a Chongqing community health service center. Questionnaire survey was conducted about general information of respondents,disease and drug use condition and attitude to deprescribing by using Australian Patients'Attitudes Towards Deprescribing questionnaire as reference. The results of questionnaire survey were analyzed statistically. The cognitive function was assessed by Hastgawa Dementia Scale. The degree of weakness was evaluated by Chinese edition of Edmonton Frail Scale. The quality of life was evaluated by Chinese edition of European Five Dimensional Health (EQ-5D) Scale. RESULTS:Totally 311 valid questionnaires were obtained,with effective recovery rate of 92.8%. Overall,311 participants were recruited,with a median age of 70 years,5 types of median disease (hypertension was most common) and 5 types of median medication (mainly calcium channel blocker and non-insulin hypoglycemic agent);54.7%(170 cases)participants took multiple drugs. The attitudes towards deprescribing were that 39.5% of respondents believed that they took too many drugs;73.9% had a desire to reduce their drugs;83.6% reported that they would be willing to reduce drugs if their doctor said it was possible. The patients taking multiple drugs preferred to use less drugs (P=0.001) and reduce current drugs (P=0.001),and were more willing to reduce drug cost by reducing drug use;they were also more worried about drug side effects than those without taking multiple drugs (P=0.005). CONCLUSIONS:The phenomenon of elderly chronic diseases patients with multiple diseases and multiple drug use are popular in community. It is necessary to simpilfy prescription. Most elderly patients would like to reduce their medications if doctors say it is possible;the patients with multiple drug use are more willing to simplify prescription than those without multiple drug use.
8.Study on islet function of SD rats in second and third trimesters of pregnancy
huan-zhen, CHEN ; xiao, WANG ; feng-ying, LI ; yun, LIU ; hong-mei, LONG ; ling, WU ; cui-ping, ZHANG ; guo, LI ; min, LUO
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(04):-
Objective To investigate the glucose tolerance and ability of insulin secretion in SD rats in second and third trimesters of pregnancy. Methods SD rats with pregnancy of 15 d were selected as experimental group(n=6),and another 6 rats of the same batch without pregnancy were served as controls(n=6).Intraperitoneal glucose tolerance trials(IPGTT) were conducted in these two groups.Rat islets were isolated after in situ collagenase digestion through pancreatic duct perfusion,islet morphology was observed by inverted phase contrast microscope,and insulin secretion was determined by radioimmunoassay. Results It was revealed by IPGTT that the levels of glucose at 30,60,90 and 120 min were significantly lower in experimental group than those in control group(P
9.Expression and Purification of Human Parathyroid Hormone Peptide(1-34) in Escherichia coli
Jian-Feng LI ; Hong-Jian XIAO ; Qiu-Yan JI ; Zhi-Hua LI ; Hai-Ting LONG ; Ling-Mei YAN ; Na LUO ; Wei-Ming XU ;
China Biotechnology 2006;0(03):-
Human parathyroid hormone peptide1-34(hPTH1-34) was highly expressed in Escherichia coli by inserting the synthesized hPTH1-34 cDNA into pThioHis, the prokaryotic expression vector. The expressed hPTH1-34 was purified by chelating sepharose immobilized metal ion affinity, reverse and filter chromatographic steps. Its purity was verified above 95% by HPLC. The quality was identified by N-terminal sequencing and MALDI-TOF-MS analysis. In vitro analysis showed the adenylate cyclase of ROS 17/2.8 cells was activated by hPTH1-34.
10.Relationship between mitochondrial DNA instability and interleukin-8 activity in gastric mucosa.
Xian-long LING ; Dian-chun FANG ; Rong-quan WANG ; Shi-ming YANG ; Yuan-hui LUO
Chinese Journal of Pathology 2003;32(1):35-37
OBJECTIVETo evaluate the relationship between mitochondrial DNA instability (mtMSI) and interleukin-8 (IL-8) activity in gastric mucosa of various lesions.
METHODSIL-8 level in gastric mucosa was assayed using ELISA method. The mtMSI was detected by PCR-SSCP techniques.
RESULTSmtMSI was observed in 11 out of 30 (36.7%) gastric cancers, 2 of 15 (13.3%) intestinal metaplasia, 2 of 10 dysplasia and 1 of 10 chronic atrophic gastritis. IL-8 level in mtMSI+ group [(76.8 +/- 3.8) pg/mg] was significantly higher than that in mtMSI- group [(48.3 +/- 3.6) pg/mg, P < 0.05].
CONCLUSIONmtMSI closely correlates with IL-8 level in gastric mucosa and is involved in gastric carcinogenesis.
DNA, Mitochondrial ; genetics ; Enzyme-Linked Immunosorbent Assay ; Gastric Mucosa ; metabolism ; pathology ; Gastritis, Atrophic ; genetics ; metabolism ; Genomic Instability ; Humans ; Interleukin-8 ; metabolism ; Metaplasia ; genetics ; metabolism ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Precancerous Conditions ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism