1.A study on the expression of CD163 in neonatal rats with Escherichia coli sepsis
Xinxian GUAN ; Shenglin YU ; Jian WANG ; Shasha GAO ; Yiping LI
Chinese Journal of Neonatology 2016;11(5):375-378
Objective To study the expression of CD163 in macrophages and sCD163 level in the serum of neonatal rat model of Escherichia coli (E. coli) sepsis. Methods A total of 72 specific-pathogen-free (SPF) neonatal rats (P7) were randomly and equally assigned into experiment group and control group. E. coli was injected peritoneally and the sepsis model was established in the experiential group while normal saline (NS) was injected in the control group. Samples were collected at 2, 4, 6, 12, 24 h and 48 h after the treatment. CD163 expression in macrophages of lung and liver tissues were tested using immunohistochemical(IHC) method, and the dynamic changes of sCD163 concentration in the serum were monitored usingenzyme-linked immunosorbent assay (ELISA) method. Results In the experiment group, CD163 expression in macrophages of lung and liver were gradually decreased at eachtime point (P <0. 001). At 2 h, CD163 expression in macrophages showed no significant differences between the two groups (P >0. 05). At 4 h and later timepoints, the differences were statistically significant (P <0. 001) . Meanwhile, sCD163 in the serum increased gradually (P <0. 01). At 2 h, sCD163 in the serum showed no significant differences between the two groups (P >0. 05). At 4 h and later timepoints, the differences were statistically significant (P <0. 001). Conclusions CD163 plays an important role in sepsis.
2.Total glucosides of paeony inhibits adriamycin-induced nephropathy in rats through blocking TLR4/NF-κB/TGF-β1 signaling
Yali ZHU ; Fengjun GUAN ; Na AN ; Shasha BAI ; Shanshan YANG
Chinese Journal of Biochemical Pharmaceutics 2015;37(7):43-46
Objective To study effect and its possible mechanism of total glucosides of paeony ( TGP ) on adriamycin-induced nephropathy rats.Methods Thirty male SD rats were randomly divided into normal group ( CON) , adriamycin-induced nephropathy group ( ADRN) and TGP-treated ADRN group (TGP).The rat nephropathy model was established by adriamycin injection.At the end of the 8th week after treatment, ELISA was used to detect the level of Cr and BUN.The values of 24 urine protein was determined by urinary protein kit.Masson staining was used to observe the fibrosis.RT-PCR was used to detect the mRNA levels of TLR4/NF-κB/TGF-β1 .Immunohistochemical method was used to detect the content of TLR4/NF-κB/TGF-β1.ResuIts Compared with the CON group, the level of Cr, BUN, 24 urine protein of ADRN group rised(P<0.01), the degree of fibrosis of ADRN group were aggravated(P<0.01), and the expressions of TLR4/NF-κB/TGF-β1 of ADRN group increased significantly.However, compared with ADRN group, the(P<0.01) level of Cr, BUN, 24 urine protein of TGP-treated rats reduced(P<0.01), the degree of fibrosis of TGP-treated rats eased(P<0.05), and the expressions of TLR4/NF-κB/TGF-β1 of TGP-treated rats decreased significantly.ConcIusion TGP retards the process of fibrosis and reduces the pathological damage in adriamycin-induced rats by down-regulating the expression of TLR4/NF-κB/TGF-β1 signaling.
3.Transplantation of bone marrow mesenchymal stem cells affects the proliferation and function of CD4+T cells in mice
Shaohong SU ; Junfeng ZHANG ; Qianru LI ; Shasha GUAN ; Ying DU
Chinese Journal of Tissue Engineering Research 2014;(1):106-111
BACKGROUND:Mesenchymal stem cells are found to have the immunoregulatory activities and a potential application prospect in the treatment of autoimmune diseases.
OBJECTIVE:To explore the mechanism of transplanting mesenchymal stems cells on the treatment of multiple sclerosis.
METHODS:The mouse mesenchymal stems cells were prepared, and injected into the al ogenic and syngenic normal mice, to detect the frequency of CD4+CD25+Foxp3+T cells in the spleen, thymus, and lymph nodes by flow cytometry, and to detect the Foxp3, transforming growth factor-β1, and interleukin-10 mRNA in the spleen, thymus, and lymph nodes by reverse transcription-PCR.
RESULTS AND CONCLUSION:Transplantation of mesenchymal stem cells on normal mice led to a significant up-regulation of CD4+CD25+Foxp3+T cells, Foxp3, transforming growth factor-β1, and interleukin-10 mRNA in the spleen, thymus, and lymph nodes both in the al ogenic and syngenic transplant groups. Transplantation of mesenchymal stem cells may be an available method in the treatment of autoimmune diseases, and CD4+CD25+Foxp3+T cell, Foxp3, transforming growth factor-β1, and interleukin-10 may be involved in this process.
4.Association between Q-1, T2 locus allele in ADAM33 gene and chronic obstructive pulmonary disease in Xinjiang Kazak, Han population
Shasha WANG ; Jian GUAN ; Shan WANG ; Fangjuan CHENG ; Xia REN
The Journal of Practical Medicine 2016;32(7):1100-1103
Objective To explore the relationship of the ADAM33 gene Q-1,T2 single nucleotide polymorphism (SNP) and suffering from chronic obstructive pulmonary disease (COPD) in Xinjiang Kazakh and Han population. Methods Peripheral blood samples to extract DNA, and the single nucleotide polymorphisms of Q-1 and T2 in ADAM33 gene were detected by SNaPshot SNP genotyping. Results Case group compared with the control group, frequencies of Q-1 locus genotypes and alleles were significant differences in Kazak (P<0.05). In patient group, there were significant differences in ADAM33 gene Q-1 locus genotypes FEV1% predicted, FEV1/FVC of clinical indicators lung function in Kazak, Han(P < 0.05). Kazak Q-1 locus AA genotype,Han GA genotype than GG genotype were significant difference.Compare Kazak AA genotype,Han GA genotype with GG genotype is more likely to cause COPD in Q-1 locus(P<0.05). In the comparison of the case and control group the two ethnic, there was no significant difference between the frequencies of T2 locus genotypes and the frequencies of Q-1,T2 the alleles (P > 0.05). There were no significant differences in T2 locus genotypes and clinical indicators of lung function FEV1% predicted and FEV1/FVC in patient group (P > 0.05). Conclusion The ADAM33 gene Q-1 locus may be related to the COPD susceptibility in Xinjiang Kazak, Han.
5.Study on the Percutaneous Permeability of Hydrocortisone Cream with Different Substrates in Diabetic Model Rats
Jianming ZHANG ; Shasha LI ; Zhuoji XIE ; Dongling GUAN ; Guofeng LI
China Pharmacy 2015;(19):2667-2669,2670
OBJECTIVE:To study the percutaneous permeability of Hydrocortisone cream with different substrates in diabetic model rats. METHODS:The Hydrocortisone O/W(oil/water)cream,water-soluble cream and oil-soluble cream were respectively prepared. Wistar rats were randomly divided into normal control group and model group. Model group was given streptozotocin(40 mg/kg)to reproduce diabetic model. Franz diffusion cell percutaneous test and HPLC were used to detect the percutaneous permea-bility rates of Hydrocortisone O/W cream,water-soluble cream and oil-soluble cream in rats of 2 groups. RESULTS:Compared with normal control group,the percutaneous permeability rates of Hydrocortisone O/W cream and water-soluble cream were obvi-ously increased,with significant difference(P<0.05);there was no significant difference in the percutaneous permeability rate of oil-soluble cream (P>0.05). CONCLUSIONS:Hydrocortisone O/W cream and water-soluble cream are easier to go through the skin of diabetic model rats,and Hydrocortisone oil-soluble cream is hard.
6.Multiplex allele-specific PCR assays for the identification of mitochondrial 12S rRNA mutations
Jing ZHENG ; Aifen YANG ; Ting ZHANG ; Qiongmin ZHANG ; Shasha GONG ; Guanghua PENG ; Yi ZHU ; Minxin GUAN
Chinese Journal of Laboratory Medicine 2011;34(7):628-632
Objective To investigate the clinical application of multiplex allele-specific PCR assays for simultaneous detection of the mitochondrial 12S rRNA A1555G and C1494T mutations associated with aminoglycoside-induced hearing impairment.Methods Three standard plasmids of different genotypes (wild-type, A1555G mutant and C1494T mutant) were constructed for templates and allele-specific primers aiming directly at wild-type and mutant of mitochondrial DNA nt1555 and nt1494 were designed for developing a multiplex allele-specific PCR technique to detect the A1555G and C1494T mutations.Then the method was applied to clinical screening of 138 non-syndromic hearing loss subjects and confirmed by DNA sequencing.Results Multiplex allele-specific PCR was successfully applied to the detection of A1555G and C1494T mutations in a cohort of 138 Han Chinese genetically unrelated hearing-loss subjects.Finally, 11(7.97%) unrelated affected subjects harbored the A1555G and C1494T mutations in the 12S rRNA gene(10 cases for A1555G and 1 cases for C1494T), which was well consistent with results of DNA sequencing [7.97%(11/138), Kappa=1.000, P<0.01].Conclusion This study indicates that the multiplex allele-specific PCR assay is useful, convenient and reliable in the detection of the A1555G and C1494T mutations, which could identify the subjects at risk and effectively prevent of aminoglycoside-induced hearing loss.
7.Association between polymorphism of S1, S2 locus allele in ADAM33 gene and chronic obstructive pulmonary disease in Xinjiang Uygur population
Ee HAO ; Jian GUAN ; Xilin XU ; Yan GAO ; Zhonghong ZHANG ; Shasha WANG ; Shan WANG
Tianjin Medical Journal 2015;(3):229-232
Objective To investigate the association between polymorphism of S1, S2 locus allele in ADAM 33 gene and chronic obstructive pulmonary disease (COPD) and lung function in Xinjiang Uygur population. Methods Blood sam?ples from 217 COPD patients and 218 healthy controls were collected. Samples of DNA was extracted, and S1, S2 single nu?cleotide polymorphism (ADAM 33) was detected by ABI SNaPshot SNP genotyping. Results There were no significant dif?ferences in the frequencies of S1 locus CC, CT, TT genotypes and C, T alleles between patient group and control group (P>0.05). There were no significant differences in the frequencies of S1 locus CC, CG, GG genotypes and C, G alleles between patient group and control group (P>0.05). In patient group, there were no significant differences in S1, S2 locus genotype and clinical indicators of lung function display, and in the FEV1%predicted and FEV1/FVC (P>0.05). Haplotype analysis showed that there were no significant differences in three kinds of haplotypes between patient group and control group ( P>0.05). Conclusion There is no significant difference in the polymorphism of S1, S2 locus allele in ADAM 33 gene and the susceptibility to COPD in Xinjiang Uygur population.
8.Human umbilical cord mesenchymal stem cells co-culture ameliorates podocytic apoptosis:a possible role of HGF
Wenwen QI ; Shasha LYU ; Gang LIU ; Jing CHENG ; Yan SONG ; Tongtong MING ; Guangju GUAN
Chinese Journal of Nephrology 2014;(12):933-938
Objective To explore the effects of human umbilical cord mesenchymal stem cells (HUC?MSCs) on podocytic apoptosis and injury induced by high glucose (HG) and the underlying mechanisms. Methods Podocytes were divided into six groups according to treatment: ⑴ normal glucose group (NG);⑵high glucose group (HG);⑶mannitol control group (NG+Ma);⑷HUC?MSC co?culture group (HUC?MSCs); ⑸ recombinant human hepatocyte growth factor treatment group (rhHGF);⑹ neutralizing antibody group(HGF?NtAb). Cytometry and Hoechst staining were used to detect the apoptosis rates. Western blot was used to measure the ratio of active PARP to total PARP and the level of Bcl?2. Immunofluorescence was used to study podocytic apoptosis and injury. Neutralizing antibody (NtAb) was used to block its function and the recombinant cytokine was added to induce its function. Results High glucose induced podocytic apoptosis in a time?dependent manner, HUC?MSCs co?culture decreased the podocytic apoptosis rate and the expression of PARP (all P﹤0.05), increased the expression of Bcl?2, prevented the reduced expression and maintained the normal arrangement of podocytic podoplanin. The rhHGF prevented podocytic apoptosis and injury similarly to HUC?MSCs, the beneficial effect of HUC?MSC decreased when blockade of HGF. Conclusions HUC?MSCs co?culture ameliorates podocytic apoptosis and injure induced by HG, probably through secreting soluble HGF.
9.The association between the polymorphisms in the IL-4-590C/T gene and children asthma disease:a Meta analysis
Xia REN ; Shasha WANG ; Shan WANG ; Fangjuan CHENG ; Chengyuan MA ; Jian GUAN
Chongqing Medicine 2017;46(5):638-641,647
Objective To investigate the relationship between polymorphism of IL-4-590C/T and susceptibility of asthma.Methods The case-control articles reporting the relationship between IL-4-590C/T polymorphism and susceptibility of asthma were collected by China National Knowledge Infrastructure,WanFang data,VIP citation databases,Pubmed,Baidu Scholar,time limits are retrieved from the building a database to January 2016.The Meta-analysis software RevMan5.0 and Stata 12.0 was applied for heterogeneity test and pooled OR calculation.Results Seven case-control studies were selected,including 1 167 cases in the asthma group and 1 101 cases in the control group.Meta-analysis showed that both-590C/T polymorphisms genotypes were significantly associated with asthma,five kinds of senotypes OR(95% CI) were CT+CC vs.TT[0.7 (0.57-0.85)],CC vs.CT+ TT [0.56(0.43-0.72)],CC vs.TT[0.46(0.33-0.64)],CC vs.CT[0.64(0.48-0.85)],C vs.T[0.45(0.27-0.77)].From subgroup analysis,genotype CC vs.CT+TT[0.50(0.35-0.72)],CC vs.TT[0.50(0.27-0.95)],CT vs.TT[0.61(0.41-0.92)],C vs.T[0.47 (0.23-0.95)] with risk correlated in Asian children asthma(P value is 0.01,0.04,0.02,0.03).Genotype CC vs.CT+TT[0.63(0.44-0.90)],CC vs.TT[0.49(0.25-0.96)],CC vs.CT[0.67(0.45-0.98)] also indicated a significant correlation between-590C/T polymorphisms of IL-4 and asthma in non-Asian children(P value is 0.01,0.04,0.04).Conclusion Current evidence suggests that the-590C/T polymorphism of IL-4 gene is associated with children asthma.
10.Association between ADAM33 gene polymorphism with chronic obstructive pulmonary disease incidence in Kazakh of Xinjiang
Shasha WANG ; Jian GUAN ; Shan WANG ; Fangjuan CHENG ; Xia REN ; Xilin XU ; Yan GAO
Tianjin Medical Journal 2015;(12):1349-1352,1353
Objective To explore correlation of Xinjiang Kazakh population who suffered from COPD with polymor?phisms of F+1,S2,T1,ST+5 locus of ADAM33 gene. Methods Blood samples (n=193) from healthy controls (Control group, n=193) and COPD patients (Case group, n=197) were detected by SNP SNaP shot. Results Comparing case group with the control group, gene frequency and allele frequency of F+1 locus were of significant differences (P<0.05). In patient group, there were no significant differences in F+1 locus genotype and in clinical indicators include lung function FEV1 predicted and FEV1/FVC (P>0.05). The gene frequencies and allele frequency of S2、T1 and ST+5 locus were not significantly differ?ent between case group and control group (P>0.05). F+1 and S2 locus were analyzed by haplotype analysis which showed that there was significant differences in Hap1 (CC) haplotype between case group and control group (P<0.05), and OR<1 indicated that its haplotype may reduce the risk of COPD . There were significant differences (P<0.05) in Hap3(TC) haplo?type between case group and control group and OR>1 revealed that its haplotype may increase the risk of COPD . The distri?bution of Hap2 (TG) and Hap4 (CG) were not significantly different (P>0.05) between the 2 groups. T1 and ST+5 locus were analyzed by haplotype analysis which showed significant differences in haplotypes between case group and control group (P<0.05). Conclusion The occurrence of COPD may be related to the polymorphism of ADAM33 gene in F+1 locus in Xinjiang Kazakh.