1.Determination of Urinary Manganese by Flame Atomic Absorption Spectrometry with Coprecipitation
Journal of Environment and Health 2007;0(07):-
0.05). Conclusion The present method has many advantages such as simplicity of operation,rapidity,better accuracy for the accurate quantitative analysis of urinary manganese.
2.Association of-866G/A polymorphism in uncoupling protein 2 gene of patients with type 2 diabetes in Nanjing
Xujun SHEN ; Dalong ZHU ; Guoyu TONG
Chinese Journal of Practical Internal Medicine 2001;0(09):-
Objective To study the assiociation of-866G/A polymorphism in the promoter of uncoupling protein 2(UCP2)gene in patients with type 2 diabetes and B-cell function in Han population in Nanjing.Methods For the case-control study,PCR-RFLP method was used to determine the distributions of allele and genotype frequencies of-866G/A polymorphism in the human UCP2 gene in 229 type 2 diabetic patients and 196 normal control subjects.All of the testees accepted oral glucose tolerance test and insulin releasing test to detect B-cell secretion function and insulin sensitivity.Results The distribution of genotype and allele frequencies of-866G/A polymorphism in the promoter of the human UCP2 gene were significantly different between type 2 diabetic patients and normal control subjects(?2=6.555,P=0.038;?2=6.363,P=0.012 respectively).The frequency of A/A genotype and A allele in type 2 diabetic were significantly higher than that in contrast(OR=1.99,OR=1.42 respectively,both P
3.AngiotensinⅡpromotes fibroblasts phenotypic transformation through down-regulating adventitial catalase expression
Kai SHEN ; Zhuoming LIN ; Shiliang CHEN ; Guoyu YUAN ; Xiaoguang LIU
Acta Anatomica Sinica 2014;(3):411-415
Objective To investigate the relationship between angiotensin Ⅱ ( AngⅡ) and transformation of vascular fibroblasts phenotype .Methods Eighteen rats were randomly assigned into the untreated group , mini-pump infusion of saline group and mini-pump infusion of AngⅡgroup which was used as the hypertension model .Their systolic pressure and vascular morphology were examined .The expression of catalase ( CAT ) and 4HNE was examined by immunohistochemistry .Western blotting was used to examine the expression of CAT of adventitial fibroblasts which were cultured by different incubation times and concentrations of Ang Ⅱ.Results Compared with untreated and mini-pump infusion of saline groups , the systolic pressure and carotid media thickness stained by HE of mini-pump infusion of AngⅡgroup were significantly higher (P<0.01).The results of immunohistochemistry showed that the expression of CAT ofAngⅡgroup was significantly lower than that of untreated group , however the expression of 4HNE of AngⅡgroupwas higher than that of untreated group (P<0.05).Furthermore, the results of Western blotting indicated that the effect of Ang Ⅱ on down-regulation of CAT function was in a dose and incubation time dependent manner .Conclusion AngⅡdown-regulates adventitial CAT expression and promotes fibroblasts phenotypic transformation which leads to pathological arterial vascular remodeling .
4.Prevalence of depression and its risk factors in type2 diabetes mellitus
Juan ZHANG ; Yan BI ; Shahmei SHEN ; Guoyu TONG ; Shiwei CUI ; Weinan YU ; Dalong ZHU
Chinese Journal of Endocrinology and Metabolism 2011;27(10):796-799
Objective To investigate the prevalence of depression and its risk factors in type 2 diabetes mellitus.Methods Beck Depression Inventory (BDI) was used to evaluate depression in 2 966 type 2 diabetes mellitus patients [ male 1 463,female 1 503,age ( 56.4 ± 11.2 ) years,diabetes duration ( 6.3 ± 5.7 ) years ].Depression criteria:≤4 points,no depression group; 5-13 points,mild depression group; 14-20 points,moderate depression group; 21 points or higher,severe depression group.Meanwhile,the demographic and metabolic data and diabetes-related health behaviors were also investigated.The risk factors associated with depression were screened by logistic regression.Results 51% patients had depression,including 38% with mild,8% with moderate,and 5% with severe depression.Compared with no depression group,depression was correlated with female sex,low-annual income,diabetes education,diabetes treatment,with insulin and peripheral neuropathy( P<0.01 ).More smoking and younger age were found in severe depression( P<0.05 ).Partial correlation analysis showed that depression in type 2 diabetes mellitus was positively correlated with female sex,diabetes education,and peripheral neuropathy ; and negatively correlated with age (P<0.05).Conditional logistic regression equation showed that gender( OR =1.37 ),age ( 20-40 years,OR =1.52 ),diabetes education ( OR =1.51 ),and peripheral neuropathy ( OR =1.87 ) were risk factors for depression.Conclusion Depression is common in type 2 diabetes mellitus.More attention should be paid to screening depression in clinical practice.
5.To investigate the relationship between P15INK4B gene promoter methylation degree and different types of MDS and prognosis
Guoyu HU ; Kui TAN ; Chaohui YUAN ; Chanjuan SHEN ; Jing LUO ; Yuxia LIU
Journal of Chinese Physician 2015;17(9):1339-1342
Objective To determine the methylation level of P15INK4B gene promoter in different types of myelodysplastic syndromes (MDS) and its correlation with its prognosis.Methods Methylation frequency of the P15INK4B gene promoter in 44 cases of MDS were determined by methylation-specific polymerase chain reaction (PCR) and pyrosequencing,and its correlation with clinical classification and characteristics of MDS were statistically analyzed.Results Frequency of P15INK4B gene promoter methylation in myelodysplastic syndromes-refractory anemia with excess blasts Ⅱ (MDS-RAEB Ⅱ) patients was (46.89 ± 15.41) %,significandy higher than that in other types of MDS (P < 0.05),but no difference in promoter methylation frequency was detected among the other types of MDS (P > 0.05) ; frequency of P15INK4B gene promoter methylation was found to be correlated with decline in platelet upon diagnosis (t =9.02,P < 0.01),but showed no significant correlation with drop of hemoglobin or leukopenia (P >0.05).As for the correlation between P15INK4B gene promoter methylation and MDS risk stratification,no significant difference was detected between the low-risk and very low-risk groups (P > 0.05),but significant differences were detected among the medium-risk,high-risk,and very high-risk groups (P < 0.05).In addition,frequency of P15INK4B gene promoter methylation was (49.21 ± 8.78)% in MDS patients that developed leukemia in the following two year,significantly higher than that in MDS patients who didn't (19.64 ± 6.24) % (P < 0.05).Conclusions P15INK4B gene promoter methylation frequency is a valuable indicator of prognosis of MDS patients.
6.Angiotensin II regulates catalase expression and promotes fibroblast phe-notype transformation through ERK1/2 pathway
Kai SHEN ; Fen CHEN ; Zhuoming LIN ; Shiliang CHEN ; Guoyu YUAN ; Xiaoguang LIU
Chinese Journal of Pathophysiology 2014;33(4):711-714
AIM:To explore the effect of extracellular signal-regulated kinase 1/2 ( ERK1/2) on the vascular adventitial remodeling in a hypertension rat model .METHODS:The rats were randomly divided into control group , mini-pump infusion of saline group and mini-pump infusion of angiotensin II ( Ang II) group as the hypertension model .The sys-tolic pressure and vascular morphology of the rats were examined .Adventitial fibroblasts were treated with Ang II , PD98059 ( ERK1/2 inhibitor) and Ang II+PD98059.The catalase ( CAT) expression in the cells was detected by Western blotting . RESULTS:Compared with control group and mini-pump infusion of saline group , the systolic pressure and carotid media thickness (stained by HE) in mini-pump infusion of Ang II group were significantly increased (P<0.01).Meanwhile, ar-tery morphology in mini-pump infusion of Ang II group had obviously changed with a significant occurrence of pathological vascular remodeling .The result of Western blotting showed that the expression of CAT in the adventitial fibroblasts treated with Ang II+PD98059 was much higher than that in the cells treated with Ang II alone (P<0.05), indicating that down-regulation of CAT induced by Ang II was restored by ERK 1/2 signaling pathway .CONCLUSION:Ang II down-regulates CAT through ERK1/2 pathway and promotes cell phenotype transformation , which lead to pathological vascular remodeling .
7.Research on antibacterial property of snake venom antibacterial peptide OH-CATH and chitosan as artificial implant coating
Siman LI ; Guoyu YU ; Guomin ZHAI ; Zhenhua GAO ; Jihong SHEN ; Yancheng PENG
Chongqing Medicine 2014;(11):1287-1289
Objective To explore the bacteriostatic efficacy of snake venom antibacterial peptide OH-CATH and chitosan as ar-tificial implant coating on escherichia coli (E .coli) .Methods The catheters and dacron patches (blank group ,chitosan group ,cef-operazone group and antibacterial peptide group) were performed the pretreatment .Then the in vitro bacteriostatic test was conduc-ted to evaluate the antibacterial activity U value on E .coli ATCC 25922 and cephalosporin-resistant E .coli on 1 d and on 7 d after putting it into plasma .Results The cefoperazone group and the antibiotic peptide group had the very powerful antibacterial activity on E .coli ATCC 25922 on 1 ,7 d ,but the antibacterial activity of the cefoperazone group was stronger than that of the antibiotic peptide group(P<0 .05);the cefoperazone group had no antibacterial activity on cephalosporin-resistant E .coli ,but the antibiotic peptide group had significantly antibacterial activity on 1 ,7 d(P<0 .01) .Conclusion Snake venom antibacterial peptide OH-CATH has very obvious antibacterial effect on the standard strain and drug-resistant strain of E .coli and its value applied in artificial im-plant coating is affirmed .
8.On-line near-infrared spectroscopy optimizing and monitoring biotransformation process ofγ-aminobutyric acid$
Guoyu DING ; Yuanyuan HOU ; Jiamin PENG ; Yunbing SHEN ; Min JIANG ; Gang BAI
Journal of Pharmaceutical Analysis 2016;6(3):171-178
Near-infrared spectroscopy (NIRS) with its fast and nondestructive advantages can be qualified for the real-time quantitative analysis. This paper demonstrates that NIRS combined with partial least squares (PLS) regression can be used as a rapid analytical method to simultaneously quantify L-glutamic acid (L-Glu) andγ-aminobutyric acid (GABA) in a biotransformation process and to guide the optimization of production conditions when the merits of NIRS are combined with response surface methodology. The high performance liquid chromatography (HPLC) reference analysis was performed by the o-phthaldialdehyde pre-column derivatization. NIRS measurements of two batches of 141 samples were firstly analyzed by PLS with several spectral pre-processing methods. Compared with those of the HPLC reference analysis, the resulting determination coefficients (R2), root mean square error of prediction (RMSEP) and residual predictive deviation (RPD) of the external validation for the L-Glu concentration were 99.5%, 1.62 g/L, and 11.3, respectively. For the GABA concentration, R2, RMSEP, and RPD were 99.8%, 4.00 g/L, and 16.4, re-spectively. This NIRS model was then used to optimize the biotransformation process through a Box-Behnken experimental design. Under the optimal conditions without pH adjustment, 200 g/L L-Glu could be catalyzed by 7148 U/L glutamate decarboxylase (GAD) to GABA, reaching 99%conversion at the fifth hour. NIRS analysis provided timely information on the conversion from L-Glu to GABA. The results suggest that the NIRS model can not only be used for the routine profiling of enzymatic conversion, providing a simple and effective method of monitoring the biotransformation process of GABA, but also be considered to be an optimal tool to guide the optimization of production conditions.
9.Genomic epitopes of Xinjiang hemorrhagic fever virus and the evaluation of its laboratory testing value
Huahua ZHANG ; Guoyu ZHAO ; Yang LI ; Shen SHI ; Ran AN ; Jiangling YUAN ; Dongliang LIU ; Jingrui DOU ; Tao LUO ; Surong SUN ; Yujiang ZHANG
Chinese Journal of Laboratory Medicine 2023;46(2):127-136
Objective:To grasp the distribution of fine antigenic epitope profiles of nucleoprotein (NP) and glycoprotein (GP) fragments of Crimean-Congo hemorrhagic fever virus (CCHFV) and to clarify the value of dominant antigenic epitopes in laboratory testing of Crimean-Congo hemorrhagic fever (CCHF).Methods:In a minimal synthetic short peptide consisting of 8 amino acids was segmentally expressed by CCHFV YL04057 strain using a modified bio-peptide synthesis method from 2014 to 2021 in the laboratory of Xinjiang University, College of Life Sciences. Using CCHFV polyclonal antibody or monoclonal antibody 14B7 (IgM) or CCHFV-positive sheep serum as antibodies, the minimal antigenic epitopes (BCEs) with antigenic activity on NP and GP fragments were identified by immunoblotting, and the obtained BCEs with sequence polymorphism were spatially clustered with CCHFV from different regions using the neighbor-joining method to determine the combination mode of BCEs with geographical correlation of regional distribution, to explore its application in establishing serological diagnosis. A prokaryotic expression plasmid (pET-32a), an E. coli expression plasmid (pGEX-KG) and a prokaryotic expression plasmid with an incomplete glutathione (GST188) tag (pXXGST-ST-1) were used to construct and express six dominant antigenic epitopes of different peptide lengths on NP fragments, and an indirect Enzyme-linked immunosorbent assay (ELISA) was established. CCHF sheep serum identified by immunofluorescence assay (IFA) was used as a control, and the specificity, sensitivity and overall compliance of the recombinant proteins with different peptide lengths of antigenic epitopes with IFA assay results were statistically analyzed. Results:CCHFV, NP and GP fragments had a total of 30 antigenically active BCEs, among which the core intermediate fragment NP2 (aa 170 th-305 th), which had a concentration of antigenic epitopes in the NP fragment, has 6 BCEs, and the NP1 (aa 1 st-200 th) and NP3 (aa 286 th-482 nd) at both ends have 9 BCEs; the Gc (aa 1 st-558 th) and Gn (aa 533 th-708 th) fragments of the GP fragment have 14 BCEs and a long antigenic peptide (AP) containing 15 amino acids, and the amino acid sequence homology of the NP fragment BCEs was 97.1% and that of the GP fragment BCEs was 89.1%. There was a significant difference ( P=0.0281, P<0.05). Among the 9 BCEs with sequence polymorphism in the GP fragment, 6 combined BCEs from GnEc1, GnE2, GnE4, GcE3, GcE6 and GcAP-4 (Ap) could cluster 15 CCHFV strains from different regions of the world into 5 geographical taxa, AsiaⅠ, AsiaⅡ, AficaⅠ, AficaⅡ and Europe. The constructs expressing PET-32a-NP (full length), PGEX-KG-NP2 (aa 170 th-305 th), pGEX-KG-NP2-1 (aa 235 th-275 th), PGEX-KG-NP2-1-1 (aa 237 th-256 th), pXXGST-1-NP2-1-2 (aa 250 th-265 th) and PGEX KG-NP2-1-3 (aa 260 th-276 th), six recombinant proteins CCHFV NP rabbit polyclonal antiserum (pAb) Western Blotting reaction positive, 33 sheep sera tested by IFA XHF as a reference, the sensitivity of the assay established by indirect ELISA using the recombinant proteins constructed from two fragments of NP2 and NP2-1 as antigens. The sensitivity, specificity and overall compliance were the best, with 73.4% (11/15) and 66.7% (10/15) for sensitivity, 100% (18/18) and 94.4% (17/18) for specificity, and 87.9% (29/33) and 81.8% (27/33) for overall compliance. Conclusion:CCHFV NP and GP are distributed with a high number of BCEs with antigenic immunoreactivity, among which the dominant antigenic epitopes are of high value in the laboratory serological diagnosis of CCHF.