1.The correlation study between the distribution of two single nucleotide polymorphism(SNP)loci on chromo-some 9p21 and the macrovascular disease in Xinjiang Uygur and Han nationality people with type 2 diabetes
Libin YANG ; Rui MA ; Jun AI ; Lingxia CHENG ; Yongjie JIN ; Ling DU ; Jun HU ; Hui CHENG
Chinese Journal of Primary Medicine and Pharmacy 2015;(18):2733-2736,2737
Objective To assessed the distribution of two single nucleotide polymorphism (SNP)loci (rs2383206.rs10757278)on chromosome 9p21 in Xinjiang Uygur and Han nationality populations,and to investigate correlation and the incidence of all cases of macrovascular disease (coronary artery disease,carotid atherosclerosis and peripheral arterial disease)and analysis of risk factors.To further study the correlation between the incidence of two single nucleotide polymorphism (SNP)loci (rs2383206.rs10757278)on chromosome 9p21 in type 2 diabetes melli-tus(T2DM)of Han and Uygur ethnic and the incidence of all cases vascular disease,then to analysis the risk factors. Methods 497 adults with T2DM who were treated in the Endocrinology department in hospital from May 2012 to April 2014 were involved in this study,including 298 Uygur patients and 199 Han patients.215 non -T2DMpatients who were treated in the Cardiology department in hospital were also involved in the study,including 93 Uighur patients and 122 Han patients.Then the total 712 patients were detectedby using PCR -SNP Stream technology to analyse rs2383206.rs10757278 loci SNP genotyping.The relevant results were compared with t test,two different genotype distribution and allele frequency were compared with χ2 test,multiple factors analysis were calculated by Logisitic regression.Results The distribution of genotype with two SNP loci had no significant difference between the patients in Uygur group and Han group (rs2383206χ2 =5.570,P =0.062;rs10757278 χ2 =2.721,P =0.257 ),and there's no significant difference between the patients with macrovascular disease and non -macrovascular disease in all patients(rs2383206χ2 =0.120,P =0.950;rs10757278 χ2 =1.027,P =0.598).Logisitic regression analysis showed that the incidence of macrovascular was significantly associated with increasing age(χ2 =28.820,P =0.000)and fatty liver(χ2 =5.210,P =0.020)in Uighur group with type 2 DM.In Han group with type 2 DM,the macrovascular was significantly associated with the increase of age (χ2 =19.980,P =0.000),elevated fasting blood glucose (FPG)(χ2 =4.070,P =0.044)and poor controlled with glycosylated hemoglobin (χ2 =4.280,P =0.040). Conclusion This study found that there's no correlation between two single nucleotide polymorphisms (SNPS)loci (rs2383206.rs10757278)on chromosome 9 p21 large with macrovascular in Uygur group and Han group.Increasing age,higher FPG and poor controlled with glycosylated hemoglobin combined with fatty liver were the risk factors for macrovascular.
2.Selection and identification of singledomain antibody against Peste des Petits Ruminants virus
Dan LIU ; Lingxia LI ; Xiaoan CAO ; Jinyan WU ; Guoyu DU ; Youjun SHANG
Journal of Veterinary Science 2021;22(4):e45-
Background:
Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Singledomain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility.
Objectives:
The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV.
Methods:
A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2.
Results:
The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 106 cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein.
Conclusions
The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.
3.Selection and identification of singledomain antibody against Peste des Petits Ruminants virus
Dan LIU ; Lingxia LI ; Xiaoan CAO ; Jinyan WU ; Guoyu DU ; Youjun SHANG
Journal of Veterinary Science 2021;22(4):e45-
Background:
Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Singledomain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility.
Objectives:
The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV.
Methods:
A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2.
Results:
The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 106 cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein.
Conclusions
The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.
4.Inhibition of caspase-1-dependent apoptosis suppresses peste des petits ruminants virus replication
Lingxia LI ; Shengqing LI ; Shengyi HAN ; Pengfei LI ; Guoyu DU ; Jinyan WU ; Xiaoan CAO ; Youjun SHANG
Journal of Veterinary Science 2023;24(5):e55-
Background:
Peste des petits ruminants (PPR), caused by the PPR virus (PPRV), is an acute and fatal contagious disease that mainly infects goats, sheep, and other artiodactyls.Peripheral blood mononuclear cells (PBMCs) are considered the primary innate immune cells.
Objectives:
PBMCs derived from goats were infected with PPRV and analyzed to detect the relationship between PPRV replication and apoptosis or the inflammatory response.
Methods:
Quantitative real-time polymerase chain reaction was used to identify PPRV replication and cytokines expression. Flow cytometry was conducted to detect apoptosis and the differentiation of CD4+ and CD8+T cells after PPRV infection.
Results:
PPRV stimulated the differentiation of CD4+ and CD8+ T cells. In addition, PPRV induced apoptosis in goat PBMCs. Furthermore, apoptosis and the inflammatory response induced by PPRV could be suppressed by Z-VAD-FMK and Z-YVAD-FMK, respectively.Moreover, the virus titer of PPRV was attenuated by inhibiting caspase-1-dependent apoptosis and inflammation.
Conclusions
This study showed that apoptosis and the inflammatory response play an essential role in PPR viral replication in vitro, providing a new mechanism related to the cell host response.
5.Diagnostic and prognostic value of combined detection of serum Lp-PLA2,NSE and S-100β in patients with carbon monoxide poisoning complicated with acute cerebral infarction
Yanpin WU ; Yanjing XU ; Lingxia DU ; Yiliang QIN ; Hongzhan ZHANG ; Yanlei PANG ; Yalin WANG
International Journal of Laboratory Medicine 2024;45(2):204-207,212
Objective To explore the value of combined detection of lipoprotein-associated phospholipase A2(Lp-PLA2),neuron specific enolase(NSE)and S-100 calcium binding protein β(S-100β)in the diagnosis and prognosis evaluation of acute cerebral infarction(ACI)in patients with carbon monoxide poisoning(CMP).Methods A total of 102 patients with CMP complicated with ACI admitted to the hospital from Jan-uary 2020 to November 2021 were selected as the study group,meanwhile,102 patients with simple CMP were enrolled as the control group.Patients in the study group were followed up for 6 months after discharge,ac-cording to the follow-up results,they were grouped into good prognosis group(60 cases)and poor prognosis group(42 cases).The serum levels of Lp-PLA2,NSE and S-100β were detected by enzyme-linked immunosor-bent assay(ELISA).The receiver operating characteristic(ROC)curve was applied to analyze the value of the combination of serum Lp-PLA2,NSE and S-100β in the early diagnosis and prognosis evaluation of patients with CMP and ACI.Results Compared with the control group,the levels of Lp-PLA2,NSE and S-100β in the study group were obviously higher(P<0.05).The ROC curve analysis results showed that the area under the curve(AUC)of the combined detection of serum Lp-PLA2、NSE、S-100β for the diagnosis of CMP complicat-ed with ACI was greater than the AUC of single detection of each indicator(P<0.001).Compared with the good prognosis group,the levels of Lp-PLA2,NSE and S-100β in the poor prognosis group were obviously higher(P<0.05).The results of ROC curve analysis showed that the AUC of the combined detection of ser-um Lp-PLA2、NSE、S-100β for the prognosis of patients with CMP complicated with ACI was greater than the AUC of single detection of each indicator(P<0.05).Conclusion The expression of Lp-PLA2,NSE and S-100β in serum of patients with CMP complicated with ACI is high,and the combined detection of the three has certain value in the diagnosis and prognosis evaluation for patients with CMP complicated with ACI.
6.The association between calcium supplementation in antenatal childbearing aged women and risk of small for gestational age among neonatal singleton in Shaanxi province
Xiangyu GAO ; Shanshan LI ; Wentao WU ; Jiaoyang DU ; Chenlu WU ; Xi JIANG ; Binyan ZHANG ; Baibing MI ; Lingxia ZENG ; Shaonong DANG ; Hong YAN
Chinese Journal of Epidemiology 2021;42(4):651-655
Objective:This study explored the association between antenatal calcium supplementation in the childbearing aged women and risk of small for gestational age infant (SGA) among singleton in Shaanxi province,China.Methods:Multi-stage random cluster sampling method was employed to collect information about pregnant women, who were pregnant and had definite outcomes, and their infants, from 30 districts (counties) in 2010 to 2013. Information was collected by face-to-face questionnaire survey. Generalized linear mixed models were employed after adjusting covariates. Dependent variable was whether single-birth neonate was SGA, and independent variable was calcium supplementation of childbearing aged women in different pregnant periods.Results:A total of 28 357 childbearing aged women was recruited in this study. The age of these women was (28.08±4.74) years old, of which, 79.28% were rural residents and 60.90% had calcium supplementation intake. There was a number of 12 810 female in singleton neonates. The neonatal birth weight and gestational age were (3.27±0.16) kg and (277.44±8.80) day, respectively. The prevalence of SGA was 11.35% in total, and 10.48% in mothers with maternal calcium supplementation and 12.70% in mothers without maternal calcium supplementation in whole antenatal period. There were statistically significant differences seen in antenatal calcium supplementation within the subgroups of maternal age (whether the mother was an advanced maternal woman), residential area, maternal occupation, maternal parity, maternal education level, and household incomes ( P<0.05). After adjusting these covariates, the risk of SGA among childbearing aged women with antenatal calcium supplementation showed 16% decreased risk ( OR=0.84, 95% CI: 0.77-0.92). Further analysis of the different antenatal periods showed that calcium supplementation during the second and third trimester had a statistically significant difference in reducing the risk of neonatal SGA ( P<0.05). Besides, subgroup analysis showed that there was a statistically significant difference between the perinatal calcium supplementation and the single-born neonates with SGA Significance ( P<0.05) in non-advanced women, those who had a low education level and moderate household economic status groups. Conclusion:The risk reduction of SGA among singleton neonates is related to calcium supplementation during antenatal period in Shaanxi province.