1.Expression of junctional adhesion molecule-1 in rat corneal tissue
Chinese Journal of Experimental Ophthalmology 2012;30(3):204-208
BackgroundJunctional adhesion molecule-1 (JAM-1) is intercellular transmembrane protein newly discovered and associated with the tight junction.Tight junction plays an important role in keeping the transparency of cornea,but there are few studies about JAM-1 in cornea tight junction.ObjectiveThis study was to determine the expression of JAM-1 in corneal epithelium,stroma,endothelium,and locate the distribution of JAM-1.MethodsThe corneas of two SFP Wistar rats were obtained,and the samples of epithelial lamella,corneal stroma and endothelium with Descemet membrane were prepared respectively for the detection of the expression of JAM-1 mRNA using reverse transcription polymerase chain reaction(RT-PCR).Primers were designed according to the genes as RGD web provided,and the objective genes were JAM-1,occludin and claudin-1.Products of PCR were examined by 1.5% agarose gel electrophoresis and assayed with GelDoc-lt UVP Imaging System.The corneal paraffin sections and stretched preparations of epithelium and endothelium of corneal sections from other two SFP Wistar rats were prepared for the examination of JAM-1protein expression and location by immunohistochemistry.The use of experimental animals followed the Statement of ARVO. Results JAM-1,occludin and claudin-1 mRNA were expressed in rat cornea epithelium,stroma and endothelium.PCR melting curves showed the limpid unimedality.The expression level of JAM-1 mRNA was similar to occludin mRNA and higher than that of claudin-1 with the highest level in the epithelium layer.Immunohistochemistry assay revealed that there was definite JAM-1 antibody staining in the cornea epithelium,stroma and endothelium layers.However,the basal layer of corneal epithelium presented with the strongest staining in comparison with stromal and endothelial layers.Stretched preparations of corneal epithelium and endothelium showed that JAM-1 protein appeared at the junction site of epithelial cells and endothelial cells.The basal layer of the corneal epithelium showed the strongest response,and the staining of corneal endothelium was extensive and diffuse. Conclusions JAM-1 is a composition of intercellular tight junction which expresses in cornea epithelium,endothelium and stroma.However,its appearance and level vary from different corneal layers.
2.The application of reduced sample number of Bergman's minimal model method in insulin resistance syndrome
Jie HONG ; Guang NING ; Xiaowei WANG
Chinese Journal of Endocrinology and Metabolism 2000;0(06):-
Objective To study the mechanism of insulin resistance in obesity, impaired glucose tolerance (IGT) and type 2 diabetes. Methods Insulin sensitivity index (SI), glucose effectiveness (S G) and insulin secretory function of ? islet cells were assessed by the reduced sample number of Bergman's minimal model method with frequently sampled intravenous glucose tolerance test in patients with obesity, IGT and type 2 diabetes as well as in the controls. Body mass index (BMI) and waist hip ratio (WHR) were determined in four groups. Results SI and S G were significantly higher in the controls than those in the IGT and type 2 diabetes patients. The S G in obesity group was nearly the same as that in the controls, but its SI was significantly lower. The area under curve 1 (AUC 1) of insulin in the controls was smaller than that in obesity and IGT (P
3.Changes in plasma monocyte chemoattractant protein-1 in patients with acute coronary syndrome and patients with restenosis after angioplasty
Jie YU ; Jieming MAO ; Guang WANG
Chinese Journal of Tissue Engineering Research 2007;0(04):-
AIM: To observe acute coronary syndrome (ACS), including acute myocardial infarction (AMI), and unstable angina pectoris (UAP) and plasma monocyte chemoattractant protein-1 (MCP-1) in patients with restenosis after angioplasty, and compared with normal controls. METHODS: Sixty-eight patients were selected from the Department of Cardiology, Third Hospital Affiliated to Peking University between December 2005 and April 2006, including 30 patients with AMI, 20 patients with UAP and 16 patients with restenosis after angioplasty. Thirty healthy people were selected simultaneously to be the controls. All subjects knew and agreed with the items. The level of blood lipid was determined in all enrolled subjects: ① Blood sample was obtained from the elbow of patients with AMI immediately at hospitalization (2-12 hours from onset). ② Blood sample was obtained from patients of UAP group and patients of restenosis after angioplasty group immediately after the hospitalization (within 24 hours of onset). ③ Blood sample was obtained from fasting subjects of the normal control group in the morning. Blood sample was centrifuged, separated of the plasma and then frozen at -70 ℃. The level of plasma MCP-1 was determined with ELISA for statistical analysis. RESULTS: A total of 98 enrolled subjects were involved in the analysis of results, and no one withdrew from the study. Comparison in plasma MCP-1 among all groups: The plasma MCP-1 in ACS group, UAP group and restenosis after angioplasty group were (166.7?46.5,149.4?54.9,119.7?25.0,89.2?26.4) ng/L respectively, and it was significantly higher in ACS group, UAP group and restenosis after angioplasty group than that in the normal control group (F =21.27,P
4.Right cheek swelling and pain for 2 weeks---Cushing′ syndrome complicated with facial infection
Tingwei SU ; Jie HONG ; Yiming GAO ; Weiqing WANG ; Guang NING
Chinese Journal of Endocrinology and Metabolism 2016;32(3):245-247
[Summary] A 61-year-old female patient was diagnosed as ACTH independent Cushing′ syndrome, right adrenal adenoma, right facial infection. The situation was well controlled with antibacterial agents, abscess incision drainage and ketoconazole therapy. The patient received partial adrenolectomy of her right adrenal gland 16 months later. This case indicated that anti-adrenal agents could be a reasonable choice when the Cushing′syndrome patients were acompanied with severe infection.
5.Role of Helicobacter pylori cheA gene in chemotaxis in vitro and colonizationin vivo
Guang CHEN ; Jie YAN ; Lihui XU ; Shenghai WU ; Xianjun WANG
Chinese Journal of Microbiology and Immunology 2010;30(11):1031-1037
Objective To determine the effect of cheA gene of Helicobacter pylori in the bacterial chemotaxis in vitro and colonization in vivo. Methods The entire cheA and cheY genes were amplified and cloned from genomic DNA of H. pylori NCTC11637 strain. Subsequently, the prokaryotic expression systems of cheA and cheY genes were generated and the target recombinant proteins rCheA and rCheY were extracted by Ni-NTA affinity chromatography. Rabbits were immunized with either rCheA or rCheY for obtaining antisera, and rCheA-IgG and rCheY-IgG in the antisera were prepared using ammonium sulfate precipitation plus DEAE-52 column chromatography. A suicide plasmid of cheA gene was constructed and then a cheA gene knock-out mutant ( cheA - ) was generated based on homologous recombinant exchange using the suicide plasmid. The cheA- mutant was identified using PCR and sequencing. The phosphorylation levels of CheA and CheY molecules of cheA - and wild-type strain were determined by using rCheA-IgG and rCheY-IgG anchoring the target proteins and protein phosphorylation detection kit. The differences of chemotaxis in vitro and colonization in vivo between cheA- mutant and wild-type strain were compared using chemotactic model and BALB/c infection model of H. pylori. Results The cheA gene knock-out in genome of cheA- mutant was confirmed by the results of PCR and sequencing. After treated with 0. 001-0. 1 mol/L HCI for 10 min, the phosphorylation levels of CheA and CheY molecules of wild-type strain were rapidly descended from ( 59.6 ±11.5) μmol and (55.5 ± 10.2) μmol to ( 10.8 ± 2.6) and (5. 5 ± 1.2) μmol (P < 0.05 ), while the phosphorylation of CheY molecule of cheA - mutant was no markedly changed with a persistent lower level ( P >0.05). The diameters [(10-20) ± (2-3) mm] of chemotactic aggregative rings of cheA- mutant were significantly less than those [(16-24) ± (2-3)mm] of wild-type strain (P <0.05). The positive isolation rate (90%) of H. pylori in gastric biopsy specimens of mice that infected with wild-type strain was remarkably higher than that (40%) of mice that infected with cheA- mutant (P <0.05). The result of fluorescence quantitative was also showed that the numbers (6.3 × 103 ±2.1 × 103 copies/mg) of H. pylori in gastric biopsy specimens of wild-type strain infected mice were significantly larger than those (8.3 × 101 ±3. 1 × 101 copies/mg) in gastric biopsy specimens ofcheA- mutant infected mice (P<0.05). Conclusion The cheA gene of H. pylori has an important role in the bacterial chemotaxis in vitro and colonization in vivo.
6.Research of pro-/anti-inflammatory cytokines in the study of coronary heart disease.
Chinese Journal of Integrated Traditional and Western Medicine 2010;30(1):87-90
The study of the relationship between inflammation, inflammatory cytokines and coronary heart disease (CHD) has become a hot spot. The disequilibrium of pro- and anti-inflammation cytokines tempts the attention abroad and domestically, and this provides a new way to prevent and treat CHD in Chinese medicine. This paper discussed the research progress of pro- and anti-inflammatory cytokines in the study of CHD.
Coronary Disease
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Cytokines
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Humans
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Inflammation
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Integrative Medicine
7.Changes of natural killer cells in peripheral blood in patients with newly-diagnosed type 1 diabetes mellitus
Minglan YANG ; Xiaofang ZHANG ; Li WANG ; Jie GAO ; Bokai WANG ; Jie HONG ; Xiaoying LI ; Weiqing WANG ; Guang NING ; Weiqiong GU
Chinese Journal of Endocrinology and Metabolism 2012;28(6):455-459
Objective To investigate the changes in nunmber and function of natural killer ( NK ) cells in patients with newly-diagnosed type 1 diabetes mellitus.Methods Cell courning was performed in peripheral blood mononuclear cell ( PBMC ) subsets in 43 cases with newly diagnosed type 1 diabetes ( T1D ),14 cases with newly diagnosed type 2 diabetes( T2D ) and 21 cases of normal controls by flow cytometry sorting.And then,isolating and collecting NK cells were performed in T1D patients and normal controls.Real time PCR was performed to evaluate the mRNA expression levels of NK cell activity related genes IFN-γ,perforin,NKp46,and NKp30 in NK cells.Results Compared to normal controls,both the proportion and the absolute counting of NK cells in PBMC from patients with T1D were significantly decreased [( 102±86 )/μl vs ( 355±264 )/μ1,P<0.01],while only the proportion of CD4+ cell were slightly increased( P<0.05 ).No statistical difference was observed regarding CD8+ T cells ( P>0.05 ).mRNA expression levels of NK cell activity related genes perforin and NKp46 in NK cells were remarkably down-regulated ( P<0.05 ),while IFN-γ and NKp30 were not changed compared with normal controls.Conclusions The reduced number and functional deficiency of NK cells may lead to the immune dysfunction in T1D and play an important role in the development of T1D.
8.Clinical characteristics and diagnostic analysis of thyrotropinoma
Xiaohua JIANG ; Jie CAI ; Weiqing WANG ; Qingfang SUN ; Jing XIE ; Shu WANG ; Guang NING
Chinese Journal of Endocrinology and Metabolism 2012;28(9):729-733
Objective To analyze clinical charaCteristics and diagnostic experience on thyrotropinoma (TSHoma).Methods Clinical characteristics,laboratory findings,diagnostic experience,and surgical outcome were summarized from 16 cases of TSHoma in our hospital from January 2006 to September 2011.Results ( 1 )Among 16 cases ( 9 male,7 female ) aged ( 51.63 ± 13.23 ) years,14 ( 87.5 % ) cases presented with hyperthyroidism and 2 were diagnosed by physical examination. None of them had exophthalmos, acropachy, or pre-tibial myxedema.( 2 ) Serum FT4 and FT3 levels were all above normal range with serum TSH ( 4.37 ± 2.77 ) μIU/ml.There were 25% (4/16) cases with positive serum thyroglobulin antibody and/or thyroid peroxidase antibody,none with positive TSH receptor antibody.TRH stimulating test was performed in 9 cases,3 were with positive response ( peak TSH level increased by 5.45-9.20 μIU/ml compared with baseline),and 6 without response (peak TSH level increased by -0.01-3.15 μIU/ml compared with baseline).TSH was suppressed to 27.15% (5.19%-99.15 % ) of the baseline in 11 cases in which somatostatin suppression test was carried out.Prolactin and insulin-like growth factor-Ⅰ levels were increased in 2 cases.MRI performed in 15 patients showed 10 cases of microadenomas and 5 cases of pituitary adenoma.A mass in nasal cavity was found in one case,where no mass was found in the pituitary.( 3 )Surgery was made in 11 cases ( 9 patients underwent transsphenoidal operation,one craniotomy,and one underwent operation via endoscopic nasal septum ). All patients (11/11) were proved to yield pituitary adenoma pathologically.Conclusion Thyrotropinoma should be considered in hyperthyroidism with unsuppressed TSH level,and dynamic tests facilitated early diagnosis.Ectopic thyrotropinoma shoull be considered when normal pituitary morphology was shown by MRI.
9.Approach to the patients with medullary thyroid cancer
Xiaohua JIANG ; Jie CAI ; Lei YE ; Shu WANG ; Yongju ZHAO ; Weiqing WANG ; Guang NING
Chinese Journal of Endocrinology and Metabolism 2012;28(5):433-438
Medullary thyroid cancer (MTC) is characterized hy the secretion of calcitonin that is derived from parafollicular cells.20%-25% of MTC are hereditary.Compared with other types of thyroid cancer,MTC is prone to recurrence,metastasis,and younger onset age.RET gene germline mutation accounts for the hereditary MTC,and somatic mutation is responsible for part of sporadic cases.A good correlation between phenotype and genotype is reported.We present in this article a case of medullary thyroid cancer patient with genetic diagnosis and treatment as well as postoperative follow-up together with RET gene screening results in her family members in order to call attention to the diagnosis and treatment of MTC.