1.Advancement of the relationship between PPARγ and asthma
Shengmei LI ; Weiping CHEN ; Chaobin SHEN
International Journal of Pediatrics 2010;37(1):23-25
Peroxidsome proliferator-activated rreceptor γ(PPARγ) has characteristics of regulation of adipocyte differentiation and lipid metabolism.In recent years,more and more evidences suggest that PPARγ plays an important role in the regulation of immune and inflammatory response.The PPARγ expression increased in airway of asthma patients, and PPARγ was involved in airway inflammation and airway hyperresponsiveneas. Recent studies have shown that PPARγ ligands may have a role in the treatment of asthma.
2.Study on the Regulatory Effects of Bombesin on the Gastrointestinal Morphology and Proliferation of Mucosa Cells in Neonatal Rabbits
Zhenbiao LI ; Shengmei WU ; Longhua QIAN
Chinese Journal of Perinatal Medicine 2000;0(04):-
Objective To investigate the regulatory effects of bombesin on the gastrointestinal morphology and proliferation of mucosa cells in neonatal rabbits. Methods Twenty four neonatal rabbits were divided into big,small dose experimental group and control group. The gastrointestinal morphology in neonatal rabbits was observed by using Video Image Digtal Analysis System and electron microscopy, and the proliferative rate of gastrointestinal epithelium cells was detected by using immunohistochemical assay. Results The villous height of duodenum were (520?76),(513?31),(379?44) ?m in three groups respectively. That in experimental group with big or small dose were significantly higher than that in control group( P
3.Screen and identification of immunodominant fragment of pORF5 plasmid protein from Chlamydia trachomatis
Zhansheng HE ; Yan ZOU ; Shengmei SU ; Wenbo LEI ; Zhongyu LI
Chinese Journal of Immunology 2016;(1):59-64
Objective:To investigate the immunogenicity of pORF5 plasmid protein,and further to screen and identify its im-munodominant domian.Methods: 10 different fragments of pORF5 gene including full length were amplified from the DNA of Chlamydia trachomatis serovar D by PCR and cloned into appropriate site of pGEX-6p vector to construct recombinant vectors after digested with BamHⅠand NotⅠrestriction endonucleases.After identification by PCR and sequencing,the recombinant plasmids were transformed into XL1 Blue E.coli to express the GST fusion proteins.ELISA and Western blot were carried out to identify the immunogenicity and immunoreaction of pORF5 plasmid protein.10 different fragments were reacted with sera from patients urogenitally infected with Chlamydia trachomatis, mouse polyclonal antibodies and mouse monoclonal antibodies of pORF5 plasmid protein with ELISA method.Results: pORF5 plasmid protein displayed strong immunogenicity and could induce a strong antibody response in human.The reactivity of human antibodies almost completely disappeared,when the native structure of pORF5 plasmid protein was de-stroyed.F6 that only lacked the N-terminal 66 amino acids was recognized by antibodies in ELISA as strongly as the whole pORF5 plasmid protein was.However,no other fragments were significantly recognized although there was a minimal reactivity of F2 and F3 with antibodies.Conclusion:pORF5 plasmid protein was an immunodominant antigen containing conformation-dependent epitope,and the C-terminal three quarters of pORF5 amino acid sequence was required for maintaining its immune dominance and conformation.The significance of the above findings lay a foundation for the further study on pORF5 protein function and vaccine development.
4.Investigation of the correlation between a group of metastasis-associated proteins and metastatic potency in breast carcinoma
Aiju FANG ; Yongsheng GAO ; Shengmei LI ; Wenjun LIU ; Bin MENG
Chinese Journal of General Surgery 2000;0(11):-
Objective To investigate the expression and the significance of a group of metastasis-associated proteins in invasive ductal breast carcinoma(IDC).Methods Tissue microarray containing 247 IDC specimens was constructed.The expressions of ?-B crystallin,CD44v6,MMP-2 and TIMP-2 were detected by immunohistochemistry,and the relation between the expression of these proteins and the clinicopathologic character was analyzed.Results(1) The expression rates of ?-B crystallin,CD44v6,MMP-2 and TIMP-2 in IDC were 70.0%,61.5%,57.5% and 57.1% respectively,and significantly higher than those of normal breast tissues(P
5.A survey of serum calcium, phosphorus, iron, zinc, and alkaline phosphatase in children from Kaschin-Beck disease areas and control areas in Qinghai Province
Shiyu LIU ; Qiang LI ; Yuxia SHI ; Zhijun ZHAO ; Shengmei LI ; Liyan SUN ; Lihua WANG
Chinese Journal of Endemiology 2015;34(7):473-475
Objective To investigate the serum levels of biochemical indexes such as calcium,phosphorus,iron,zinc,and alkaline phosphatase of children in Kaschin-Beck disease areas and control areas in Qinghai Province.Methods According to the results of Kaschin-Beck disease monitoring in Qinghai Province,Tangnaihai Township of Xinghai County and Gandu Town of Hualong County were chosen as Kaschin-Beck disease areas and Qushian Township of Xinghai Country was chosen as a control.Children aged 7 to 15 in boarding schools in these areas were chosen as the study subjects in June 2014.Serum alkaline phosphatase level was determined by enzyme colorimetry and the serum iron,zinc,calcium and phosphorus levels were determined by colorimetry.Calcium to phosphate ratio was calculated at the same time.Results Fifty-nine qualified serum samples of children were sampled in Kaschin-Beck disease areas and 45 in control area.Serum alkaline phosphatase levels in Kaschin-Beck disease areas and control area were (311.34 ± 85.31) and (264.09 ± 73.44)U/L,respectively,and the difference was statistically significant (t =-2.97,P < 0.05).Children's serum phosphorus levels in Kaschin-Beck disease areas and control area were (1.62 ± 0.17) and (1.43 ± 0.13)mmol/L,respectively,and the difference was statistically significant (t =-6.29,P < 0.05).Calcium to phosphorus ratio were 1.62 ± 0.17 and 1.82 ± 0.21,respectively,and the difference was statistically significant (t =5.53,P < 0.05).Compared with the control area,the level of children's serum zinc was (19.96 ± 1.70) and (20.59 ± 2.45)μmol/L;the level of serum calcium was (2.59 ± 0.11) and (2.57 ± 0.11)mmol/L;and the level of serum iron was (15.06 ± 7.02) and (17.01 ± 6.70)μmol/L.The differences between these three biochemical indicators were not statistically significant between Kaschin-Beck disease areas and control area (t =1.39,-0.64,1.44,all P > 0.05).Conclusion In Kaschin-Beck disease areas in Qinghai Province,the level of children's serum alkaline phosphatase and phosphorus have increased markedly and the calcium to phosphate ratio has decreased obviously.
6.Analysis of differential protein expression profile of HeLa cells stably transfected with Chlamydia trachomatis pORF5 gene
Wenting DAI ; Zhansheng HE ; Shengmei SU ; Zhou ZHOU ; Chaoqun CHEN ; Zhongyu LI
Chinese Journal of Microbiology and Immunology 2017;37(1):6-13
Objective To analyze the protein expression profile of HeLa cells transfected with pORF5 gene of Chlamydia trachomatis. Methods A lentiviral expression vector containing pORF5 gene was constructed. The lentiviral expression vector and helper plasmids were co-transfected into 293T cells to construct the recombinant lentivirus, which was used to infect HeLa cells. HeLa cells transfected with pORF5 gene and control HeLa cells were sorted out by flow cytometry. The isobaric tags for relative and absolute quantitation ( iTRAQ) approach combined with nano-liquid chromatography-tandem mass spec-trometry ( NanoLC-MS/MS) analysis was performed to understand protein expression profiles and to iden-tify and quantify the differentially expressed proteins in the pORF5-transfected HeLa cells ( pORF5-Hela) and the control HeLa cells. Quantitative real-time PCR ( qRT-PCR ) and Western blot analysis were performed to detect the expression of some proteins at mRNA and protein levels, respectively. Results HeLa cell line stably transfected with pORF5 gene and control HeLa cell line were constructed successful-ly. Totally 314 proteins were differentially expressed between the pORF5-HeLa and control HeLa cells, 159 of which showed increased expression and the other 155 showed decreased expression in pORF5-HeLa cells. The differentially expressed proteins were involved in many processes, such as metabolic process, immune response, biological adhesion and so on. Results of qRT-PCR showed that the expression of HIST1H1C(histone H1. 2C), HBA1(hemoglobin subunit alpha), PARK7(parkinson disease protein 7), HMGB1(high mobility group protein B1) and HMGB2 at mRNA level in pORF5-HeLa cells were up-regulated, while the expression of CLIC1 ( chloride intracellular channel protein 1 ) , KRT7 ( typeⅡ cy-toskeletal 7), SFN(14-3-3 protein sigma) and CDKN2A(cyclin-dependent kinase inhibitor 2A) were down-regulated. Western blot analysis confirmed the enhanced expression of HMGB1 and PRAK7 at pro-tein level. The results of qRT-PCR and Western blot analysis were consistent with proteomic data. Con-clusion Expression profiles for differentially expressed proteins between pORF5-HeLa and control HeLa cells were established successfully. The differentially expressed proteins regulated by pORF5 gene were found to be related to cell metabolism, proliferation, adhesion and so on, suggesting that pORF5 might promote the growth and proliferation of Ct by regulating protein expression and biological behavior of host cells.
7.Relationship between transforming growth factor-β1 and interleukin-10 in breast milk and allergic diseases in infants
Jianxing ZHU ; Liya ZHANG ; Huajun LI ; Tongxin CHEN ; Yazhong ZHU ; Shengmei WU
Chinese Journal of Perinatal Medicine 2010;13(5):364-370
Objective To verify the relationship between transforming growth factor-β1(TGF-β1) and interleukin-10 (IL-10) in breast milk and allergic diseases development in infants. Methods Totally 191 mothers (99 allergics and 92 controls) and their full-term newborns participated in this prospective study on development of children atopy. Maternal blood, cord blood, colostrum and mature milk were assayed for TGF-β1 and IL-10 by enzyme-linked immunosorbent assay. Infants underwent pediatrician evaluation for allergic diseases at six months old. Concentrations of TGF-β1 and IL-10 from allergic and non-allergic mothers and prevalence of allergic diseases of infants were compared. Results The level of IgE in allergic mothers was 30 750 IU/L(6600-410000 IU/L),lower than that in non-allergic mothers[50000 IU/L(7100-610000 IU/L)](Z=-3. 444,P=0. 001).No difference in the concentration of TGF-β1, IL-10 and IgE in mature milk was observed between allergic and non-allergic mothers. TGF-β1, IL-10 and IgE levels in colostrum of allergic mothers were 2300 pg/ml(620-7000 pg/ml), 12. 8 pg/ml(7.5-560.0 pg/ml)and 7000 IU/L(5100-56000 IU/L),significantly higher than those in non-allergic mothers[1830 pg/ml(1240-9400 pg/ml), 11. 1 pg/ml (7. 2-630.0 pg/ml)and 6700 IU/L(5200-35000 IU/L)] (Z=-2. 215, -2. 730 and -2. 706,P<0.05).In both allergic and non-allergic mothers, TGF-β1 and IL-10 levels in cord blood were higher than those in maternal blood, while IgE was lower. TGF-βl and IL-10 and IgE levels in colostrum were higher than mature milk(P<0.05). At six months old, the prevalence of allergic diseases of infants from allergic mothers(59. 6%, 59/99) was significantly higher than those from non-allergic mothers (21. 7%, 20/92)(x2= 28. 177, P= 0. 000). The prevalence of allergic diseases of infants who completed two weeks' colostrum-fed after birth (44.5 %, 73/164) was significantly higher than those who did not (22.2%,6/27)(x2 =4. 749,P-=0. 029). Conclusions High concentration of TGF-βl and IL-10 in colostrum does not show any protective effect against allergic diseases in infants. The prevalence of allergic diseases of colostrum-fed infants is significantly higher than non colostrum-fed infants, showing that colostrum-fed might play a role in allergic diseases development.
8.Purification and immunological characteristics of monoclonal antibody 2H4 against Chlamydia trachomatis pORF5 plasmid protein
Zhongyu LI ; Yimou WU ; Qiulin HUANG ; Shengmei SU ; Zhou ZHOU ; Chaoqun CHEN ; Hui ZHOU ; Guangming ZHONG
Chinese Journal of Microbiology and Immunology 2011;31(11):1041-1045
ObjectiveTo purify and characterize the monoclonal antibody (McAb) against Chlamydia trachomatis pORF5 plasmid protein.Methods The hybridoma cells stably secreting specific McAb against pORF5 were cultured in a large scale,and protein G purification by affinity chromatography was used to purify 2H4 McAb.ELISA was used to determine the antibody titer,and identify McAb isotype.Immunofluorescence assay (IFA) and Western blot were performed to detect McAb specificity.Results The purity of 2H4 antibody was 93%,the titer reached 1:1024,and 2H4 McAb was identified to belong to IgG2a isotype,2H4 McAb reacted strongly with the GST-pORF5 fusion protein and endogenous pORF5 protein expressed by Chlamydia trachomatis serovar A,D,L2,Chlamydia muridarum ( MoPn ),Chlamydia psittaci 6BC,but not other chlamydial plasmid proteins and Chlamydia pneumoniae(Cpn) AR39 strain.Conclusion2H4 McAb against pORF5 protein was successfully purified with a high titer and specificity which lay a foundation for further study on pORF5 protein structure and function.
9.Volume kinetics of lactated Ringer's solution during induction of epidural versus general anesthesia in elderly patients
Jianhong XU ; Yuhong LI ; Xiongxin ZHANG ; Fangping BAO ; Yan XIANG ; Shengmei ZHU
Chinese Journal of Anesthesiology 2013;33(11):1376-1379
Objective To compare the volume kinetics of lactated Ringer' s solution during induction of general and epidural anesthesia in elderly patients.Methods Forty ASA Ⅰ or Ⅱ patients,aged 66-86 yr,weighing 45-86 kg,undergoing elective surgery,were studied.In epidural anesthesia group (n =20),lactated Ringer' s solution was infused intravenously starting from 10 min before epidural anesthesia was performed with local anesthetics.In general anesthesia group (n =20),lactated Ringer' s solution 1000 ml was infused intravenously over 60 min starting from 20 min before induction of anesthesia.Arterial blood samples were obtained every 5 min for measurement of hemoglobin concentrations.The plasma dilution,volume increase,and volume expansion efficacy were calculated.Results The plasma dilution,volume increase and volume expansion were significantly higher at 30-60 min of lactated Ringer' s solution infusion in general anesthesia group than in epidural anesthesia group (P < 0.05 or 0.01).Conclusion The volume expansion of lactated Ringer' s solution is greater in elderly patients during induction of general anesthesia than that during induction of epidural anesthesia.
10.The expression of CD147 and PTEN in non small cell lung cancer and their correlation with clinical pathological factors
Shuguang LIU ; Zhenning ZOU ; Hongmei MA ; Xingen WANG ; Jing LI ; Shengmei XU
Chongqing Medicine 2016;45(19):2638-2640,2644
Objective To analyze the expression of extracellular matrix metalloproteinase inducer (CD147)and phosphatase and tensin homolog deleted on chromosome ten(PTEN)in non small cell lung cancer (NSCLC) ,and to explore the correlations be‐tween expressions of CD147 and PTEN and those with clinicopathological factors .Methods The expressions of CD147 and PTEN proteins in tissues of 64 cases of patients with NSCLC and 10 cases of normal paracancerous tissues were determined by using im‐munohistochemical SP method .The correlations between expressions of CD147 and PTEN with clinicopathological factors were ana‐lysed ,as well .Results The expression of CD147 in NSCLC tissues(75 .00% )was significantly higher than that in paracancerous tissues(0 .00% ,P<0 .05) .The expression of CD147 was strongly associated with degrees of differentiation ,lymph node metastasis and TNM stage(P<0 .05) .The expression of PTEN in NSCLC tissues (32 .81% )was significantly lower than that in paracancerous tissues(80 .00% ,P<0 .05) .Expression of PTEN was strongly associated with TNM stage (P<0 .05) .Spearman correlation analy‐sis shown that CD147 expression was negatively correlated with PTEN expression (r= -0 .442 ,P<0 .05) .Conclusion The abnor‐mal expression of CD147 and PTEN might play an important role in the malignant progression of NSCLC .