2.Association Study of NOD2 Gene and Coronary Artery Disease Based on Optimized DNA Ligase Chain Reaction
Yang LI ; Xi YANG ; Xiaoli TIAN
Chinese Circulation Journal 2017;32(6):569-574
Objective: Based on optimized method of DNA ligase chain reaction in medium/low throughput genotyping, we assessed the relationship between NOD-like receptor genes NOD1, NOD2 and coronary artery disease (CAD) occurrence. Methods: A multiplex PCR was conducted to enrich DNA template; probe design, annealing temperature, time and number of circulation of PCR were opfimizecl for allele specific ligation; allele specific products were identified by fluorescence PCR and capillary electrophoresis; the accuracy was verified by Sanger sequencing. Single nucleotide polymorphisms (SNPs) on NOD1 gene and NOD2 gene were examined in 1555 CAD patients and 1887 control subjects; the relationship between SNPs and CAD occurrence was studied. Results: Based on optimized PCR condition and allele specific probe design, 30 allele loci genotyping can be genotyped by 10ng DNA template at one time. Association study presented that rs751271 and rs1861759 on NOD2 gene were related to non-hypertensive CAD, allP<0.05; with Bonferroni correction, such correlation was still significant, allP<0.05. Conclusion: We optimized DNA ligase chain reaction and established a novel high accuracy, low cost method for the demand of medium/low throughput genotyping in clinical molecular diagnosis. With this method, we identified that rs1861759 and rs751271 on NOD2 gene were associated with non-hypertensive CAD.
3.Change of depression-like behavior in chronic alcoholism and withdrawal model, and co-mechanism of depression and chronic alcoholism in mice
Xi JIANG ; Furong TIAN ; Yingzheng ZHAO
Chinese Journal of Pathophysiology 2016;32(2):296-301
AIM: To investigate the behavior of depression in chronic alcoholism and withdrawal model of mice, and to explore the co-mechanism of alcoholism and depression.METHODS: A novel model of chronic alcoholism was constructed in this study.The animals were divided into normal control group, and alcohol 7 d, 14 d, 21 d and 28 d groups.The mice were given alcohol preference test on the 6th, 13th, 20th and 27th days.After the test, alcohol were withdrawn for 1 d, then the next day the mice were given behavior test of depression.After the test, the mice were sacri-ficed.The contents of 5-hydroxytryptamine (5-HT) and norepinephrine (NE) were detected by HPLC.The expression of cAMP response element-binding protein ( CREB) and brain-derived neurotrophic factor ( BDNF) was detected by Western blot.RESULTS:The mice showed an obvious drinking phenomenon, and the immobility time of forced swimming test and tail suspension test was significantly increased, with increasing drinking days and withdrawal times.5-HT level in 7 d group mice only increased in frontal cortex (P<0.05).However, compared with control group, 5-HT levels in hippocampus and cortex were decreased on the 21th and 28th days (P<0.01).NE levels in 21 d and 28 d groups were decreased in hippo-campus and frontal cortex (P<0.05), and no significant change was observed in 7 d and 14 d groups.The protein levels of p-CREB and BDNF were significantly decreased in hippocampus and frontal cortex of 12 d and 28 d groups (P<0.05), and no significant change was observed in 7 d group and 14 d group.CONCLUSION:The co-mechanism of alcoholism, withdrawal and depression is related to 5-HT.5-HT-cAMP-CREB-BDNF signaling pathway may be a common mechanism for alcoholism and depression.
4.Expressions of GLP-1R and CK17 in papillary thyroid cancer and clinical significance
Chongqing Medicine 2017;46(7):919-922
Objective To investigate the expressions of glucagon-like peptide-1 receptors (GLP-1R) and cytokeratin17 (CK17) in papillary thyroid cancer tissue and clinicopathological significance.Methods The immunohistochemistry and immunoblotting method were employed to detect the expressions of GLP-1R and CK17 in normal thyroid tissues (control group,50 cases) and papillary thyroid cancer tissue (observation group,80 cases).Then the correlation between the expressions of GLP-1R and CK17 with pathological factors of thyroid cancer was analyzed.Results Low expression of GLP-1R was found in 22 cases (44.00 %) of the control group,while GLP-1R was expressed in 77 cases(96.25 %) of the observation group,and the difference between both groups was statistically significant(P<0.01).The expression of CK17 was positive in 8 cases(16.00%) of the control group,while 77 cases(96.25 %) in the observation group were CK17 expression positive,and the difference between both groups was statistically significant(P<0.01).The difference between GLP-1 positive expression and clinicopathologic factors had no statistical significance(P>0.05).The difference between CK17 expression with sex and age had no statistical significance (P>0.05),and was obviously related with the tumor diameter,clinical stages and cervical node metastasis of thyroid neoplasms(P<0.05).Conclusion GLP-1R and CK17 all are highly expressed in papillary thyroid cancer tissue and CK17 expression is obviously related with the tumor diameter,clinical stages and cervical node metastasis of papillary thyroid cancer.
5.Sperm DNA fragmentation index and the success rate of IVF/ICSI.
Di XI ; Yun CHEN ; Yu-tian DAI
National Journal of Andrology 2016;22(1):77-81
Sperm DNA fragmentation index (DFI) refers to the percentage of DNA strand breaks in the total sperm. Many studies suggest that elevated DFI can lead to male infertility and early spontaneous abortion. High-DFI patients are more likely to fail in assisted reproduction and preliminary treatment or prevention methods have been developed for this population. This review focuses on the impact of DFI on clinical pregnancy outcomes and progress in the studies of its treatment.
Abortion, Spontaneous
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Chromatin
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DNA Fragmentation
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Female
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Humans
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Infertility, Male
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Male
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Pregnancy
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Pregnancy Outcome
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Sperm Injections, Intracytoplasmic
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Spermatozoa
6.The extraintestinal manifestation of inflammatory bowel disease:clinic study of 201 cases
Xi WU ; Xinguang LIU ; Yu TIAN
Chinese Journal of Practical Internal Medicine 2001;0(03):-
Objective To study the occurrence of the extraintestinal manifestations of inflammatory bowal disease.Methods By the retrospective study,the data of 201 IBD patients was analysed.Results A total of 21.43%of patients with ulcerative colitis(UC) and 15.79% of patients with Crohn's disease(CD) showed at least one kind of EMIs.In UC patients with EIM,the activity of disease was active phase in 89.74% and remassion phase in 10.26%.Tree CD patients were all in active phase. Musculoskeletal manifestation was the most frequent EIM,the second was dermatological manifestation,EIMs involving the genitourinary、thyroid、hepatobiliary were rarely. EN and/or PG and arthritis were more frequent coexisting other EIMs.Conclusion The prevalence of EIM was not rarely.EIM was most common among female and young patients.Arthritis and dermatological manifestations were most frequent of all assessed.The patient could have multiple EIMs and there was a tendency that EIM was more likely among patients in active phase、severe severity and with pancolitis.
7.Effect of erlotinib on renal injury in rats with STZ-induced diabetic nephropathy
Tian HUANG ; Xi CAI ; Ling ZHONG ;
Chinese Journal of Pathophysiology 2017;33(8):1460-1466
AIM: To investigate the effect of epidermal growth factor receptor (EGFR) inhibitor erlotinib on kidney injury in diabetic nephropathy (DN) rat and the underlying mechanism.METHODS: The rat model of DN was induced by intraperitoneal injection of streptozotocin (STZ) at dose of 55 mg/kg.One week after STZ injection, the rats with blood glucose level exceeding 16.7 mmol/L were identified as diabetic.Diabetic rats were randomly divided into 2 groups: STZ group and STZ+erlotinib group.In addition, the normal rats were used as control group.The rats in STZ+erlotinib group were treated with erlotinib at 100 mg·kg-1·d-1 for 4 weeks(5th~8th week).The fasting blood glucose (FBG), serum creatinine (SCr) and 24 h urine protein were measured.The pathological changes of the kidney were observed by HE staining and Masson staining.The protein levels of EGFR, p-EGFR, transforming growth factor β1 (TGFβ1), Smad2/3, p-Smad2/3, collagen Ⅳ (ColⅣ) and fibronectin in the kidney tissues were determined by Western blot.The reactive oxygen species (ROS) level and malondialdehyde (MDA) content in the renal tissues were futher analyzed.RESULTS: Compared with control group, the levels of FBG, 24 h urine protein and Scr were significantly increased in STZ group (P<0.01).Compared with STZ group, the levels of FBG, 24 h urine protein and SCr in STZ+erlotinib group were markedly decreased (P<0.05).In additon, the glomerular structure was restored to normal, the proliferative degree of mesangial cells markedly attenuated, and the epithelial cells were in alignment in STZ+erlotinib group.Moreover, erlotinib significantly inhibited the protein levels of p-EGFR, TGFβ1, p-Smad2/3, ColⅣ and fibronectin in the kidney tissues of STZ rats.In addition, erlotinib also significantly inhibited the levels of ROS and MDA in the kidney tissues of STZ rats.CONCLUSION: Erlotinib ameliorates STZ-induced diabetic nephropathy possibly through inhibiting the activation of EGFR/TGFβ1-Smad2/3 signaling pathway in association with suppression of fibrosis and oxidative stress.
8.Current situation and prospect of multidisciplinary team in thyroid cancer
Wen TIAN ; Hongqing XI ; Zheng WAN
Chinese Journal of Endocrine Surgery 2021;15(1):1-4
With the increasing number of complex and difficult thyroid cancer patients, the traditional medical model is difficult to solve the problem of diagnosis and treatment of patients optimally. In recent years, multidisciplinary team (MDT) model is applied clinically. For complex and difficult thyroid cancer diseases, the model can play a multidisciplinary advantage and provide more personalized and accurate diagnosis and treatment scheme for patients. The diagnosis and treatment of complex and difficult thyroid cancer involve thyroid surgery, otolaryngology, thoracic surgery, anesthesiology, endocrinology, nuclear medicine, radiotherapy, oncology, ultrasound, imaging, pathology and other disciplines. Based on the important role of MDT in diagnosis, treatment, nursing, postoperative management and clinical research of thyroid cancer, as well as the challenges and difficulties faced in implementation of MDT, the status and progress of multi-disciplinary comprehensive diagnosis and treatment of thyroid cancer will be systematically elaborated in the paper.
9.Preventive effect of Senecio cannabifolius Less.Ⅱon perfluoroisobutylene inhalation-induced acute lung injury in rats
Jiadan SUN ; Xiaobo WANG ; Shuang JIANG ; Ronggang XI ; Ye TIAN
Journal of International Pharmaceutical Research 2014;(4):444-448,472
Objective To preliminarily investigate the effect and possible mechanisms of Senecio cannabifolius Less.Ⅱ(FHC-Ⅱ) on perfluoroisobutylene (PFIB) inhalation-induced acute lung injury. Methods Totally 156 rats were randomly assigned to three groups: the control group, the PFIB group and the FHC-Ⅱ prevention group, with 32, 62 and 62 rats in each group respectively. The FHC-Ⅱprevention group were given FHC-Ⅱthree times per day at the dosage of 340 mg/kg before PFIB exposure. 1 h after the last time of FHC-Ⅱ administration, the FHC-Ⅱ prevention group were exposured to gaseous PFIB (0.2 mg/L) for 10 minutes in a static whole-body exposure inhalation system. The survival rate of the rats were recorded at 1, 2, 4, 8, 16, 24, 48 and 72 h post PFIB exposure;the lung index and total protein content in bronchoalveolar lavage fluid (BALF) were measured at 1 h, 2 h, 4 h, 8 h, 16 h and 24 h; IL-1β and IL-8 in sera were assayed by enzyme-linked immunosorbent assay (ELISA) at 1, 2, 4, 8 and 16 h post PFIB exposure and the histopathological examination of the lung tissue was performed at 8 h post PFIB exposure. Results FHC-II significantly reduced the content of the total protein in BALF, lung index and the levels of IL-1β and IL-8 in aera as well, and dramatically alleviated the histopathological changes in the lung tissue. Conclusion FHC-Ⅱ demonstrates some preventive effect on PFIB inhalation-induced acute lung injury in rats.
10.Research of the effect in intermedin on cell proliferation repair of renal tubular cell hypoxia-reoxygenation injury
Yanhong WANG ; Jihua TIAN ; Xiaole SU ; Xi QIAO ; Rongshan LI
Chinese Pharmacological Bulletin 2015;(4):482-486,487
Aim To evaluate the effect of intermedin (IMD)on cell proliferation and regeneration in rat tu-bular epithelial cell line (NRK-52E)that was subjec-ted to hypoxia-reoxygenation (H/R)injury.Methods The NRK-52E cells were divided into control group and three model groups (H/R,H/R +primitive vec-tor,H/R +IMD vector).The content of LDH was de-tected to observe the influence of IMD on H/R injury. The cell proliferation was detected by MTT.The cell cycle was detected by flow cytometry.Real-time PCR and western blotting were used to determine mRNA and protein levels.Results ① In comparison to the con-trol,H/R treatment decreased the cell viability and in-creased LDH activity (P <0.01 );in contrast,com-pared to H/R,IMD treatment ameliorated cell viability (79.1 5 ±1 .421 % vs 61 .22 ±1 .63%,P <0.05)and decreased LDH activities by 33.85% (P <0.01 ).②The proliferation of NRK-52E cells was significantly in-hibited by H/R treatment.In comparison to the con-trol,H/R treatment of NRK-52E cells increased the proportion of cells in the G0 /G1 phase but decreased the proportion of cells in the S and G2 /M phases. Moreover,the over-expression of IMD resulted in S and G2 /Mphase redistribution and the accumulation of G2 /M-phase cells.The real-time PCR and western blotting results indicated that the mRNA and protein expression levels of cyclin D1 ,CDK4 and p57 were increased in H/R-treated cells.IMD further stimulated this up-reg-ulated expression of cyclin D1 ,CDK4 and decreased the expression of p57 in NRK-52E cells.④Cyclin D1 had a predominantly nuclear localization in NRK-52Ecells,although cytoplasmic localization was also ob-served.Conclusion The study shows that the over-expression of IMD may promote renal cell proliferation and regeneration after renal tubular cell H/R injury via the up-regulation of cyclin D1 ,CDK and the down-reg-ulation of p57.