Background The orbital fibroblasts (OFs) in thyroid associated ophthalmopathy (TAO) play important roles in the proliferative and inflammatory response.Seeking the drug which inhibit OFs growth is of a vital significance for the prevention and treatment of TAO.Research documented that colchicine has an anti-fibrosis effect.But its influence on OFs of TAO patient is few known.Objective This study was to investigate the effect of colchicine on growth and apoptosis of OFs in vitro.Methods The retroobital connective tissue was obtained form 3 TAO patients and cultured using explant method.OFs were passaged and identified by immunochemistry,and 3-8 genetaions of cells were used in the study.Colchicine at the concentrations of 1 × 10-8,1 × 10-7,1 × 10-6,1 × 10-5,1 ×10-4 mol/L was added into the RPMI 1640 with 10％ fetal bovine serum(FBS) to incubated the cells for 24,48 and 72 hours respectively,and only RPMI 1640 was used to culture the cells as the control group.Cell counting kit-8 (CCK-8)was used to detect the absorbance value (A450) of OFs for the evaluatuion of OFs and the inhibitory rate of colchicine to OFs.The colchicine of 1 ×10-6,1 ×10-5,1 × 10-4 mol/L was added into the culture medium for 48 hours,and then the apoptotic rate of the cells and the cell percentage in various cellular cycle was assayed by flow cytometry(FCM).The expression of transforming growth factor-β (TGF-β)in the cells was detected by immunochemistry to assess the influence of colchicine on the serection of the cells.Results Cultured cells showed the spindle-like in shape and the cell number was significantly increased with the incubation time.After incubated with 1 × 10-4,1 × 10 5,1 × 10-6,1 ×10-7,1 × 10-8 mol/L colchicines,the A450 values were gradually reduced with the increase of the concentrations of colchicine(F ion =62.004,P＜0.05),and significant differences were found between different contrations of colchicine groups(all P＜0.05).Aslo,gradually declined A450 values of the cells were seen with the lapse of culture time among the groups(Ftime =459.582,P＜0.05).The inbitory rate of colchicine to the cells was elevated with the increase of concentrations.The apoptotic rates of the cells were (1.73 ± 0.15) ％,(21.04 ± 4.56) ％,(31.84 ±6.21)％and(35.32±5.56)％ in the control group and 1 × 10-6,1 × 10-5,1 × 10 4 mol/L colchicine groups respectively,with statistically significant difference among the 4 groups (F =83.905,P＜0.05).With the increase of concentrations of colchicines,the cell percentage in G2 +M phase lessened gradually,showing significant difference among the control group and the 1 × 10-6,1 × 10-5,1 × 10-4 mol/L colchicine groups (F =20.443,P＜0.05).The expression of the TGF-β in the cells was (97.60± 2.09) ％ in the control group,and that in the 1 × 10-4 mol/L colchicine group was (44.43 ± 3.96) ％,presenting a significant difference between them (t =65.330,P ＜ 0.05).Conclusions Colchicine can induce apoptosis of OFs and inhibit the prolilferation of OFs in a time-and dose-dependent manner probably by decreasing the TGF-β secretion

Objective To investigate clinical significance of proinsulin and true insulin in obese children with impaired glucose tolerance (IGT).Methods There were 21 IGT and 52 normal glucose tolerance (NGT) children. Control cases were 40 normal children. The levels of serum fasting proinsulin,true insulin,insulin,c-peptide and glucose were measured in all the subjects.Results 1.Levels of fasting proinsulin,c-peptide, glucose, insulin, true insulin and homeostasis insulin resistance in obese children with IGT showed significant difference compared with NGT (P

Adolescent ; Blood Vessels ; pathology ; Female ; Femur Head ; blood supply ; pathology ; Femur Head Necrosis ; pathology ; Humans ; Male

Purpose:To evaluate the differences in expression of carcinoembryonic antigen mRNA (CEA mRNA) in various tissues and its use in the diagnosis of digestive tract carcinoma.Methods:There were 35 patients with digestive tract carcinoma and 12 patients with non carcinoma disease in the study. The expression of CEA mRNA in the digestive tract tissues specimens was detected with nested reverse transcriptase polymerase chain reaction(RT PCR). A contrast test was detection of carcinoembryonic antigen (CEA) in blood sepcimen with enzyme linked immunosorbent assay (ELISA). Results:The expression of CEA mRNA in cancer tissues with RT PCR was 29 (82.86%) with positive results and that of CEA in blood with ELISA was 15 (42.86%) in the patients with digestive tract carcinoma. The difference in positive results was significant ( P

Objective To investigate the diagnostic significance of endoscopic findings in Henoch-Schonlein purpura(HSP),especially when abdominal pain preceded the cutaneous lesions.Methods The clinical data and gastroscopic findings in 37 cases of children with HSP were studied and analysed retrospectively in order to detect the pathological changes in the stomach and duodenum mucosa.The biopsy was taken in the pathological changeing place,and the relationship between clinical and endoscopic findings was analyzed.Results Detection rate of the pathological changes in the stomach and duodenum mucosa was 62.2%,31.3% of which experienced only cutaneous lesions,100% of which presented the acute abdominal pain.Three patients were not checked up the pathological changes.Of them,1 had arthritis,2 had Henoch-Schonlein nephritis.Characteristically endoscopic findings in the stomach and duodenum mucosa were found.The endoscopic findings included anabrosis,hyperemia,edema and hemorrhage.Conclusions Detection rate of the pathological changes in the stomach and duodenum mucosa is higher.Endoscopy is very helpful to the early diagnosis of HSP in children,especially abdominal pain presented firstly.

Objective To describe the genetic epidemiologic features of alopecia areata (AA) patients in China and to presume the possible genetic mo del of AA.Methods A case-controlled study of 1032 AA patients was performed to analyze the effect of genetic factors on the liability to AA.Complex segreg ation and heritability analysis were performed using Falconer's method and SAGE-REGTL programs.Results The mean age of onset was 28.98 ? 13.43 years.The d ifference in the mean age of onset was not significant between males and females.A total of 82.6 percent of patients experienced their first episode of AA befo re the fourth decades of life.A positive family history of AA was obtained in 8 7 patients (8.43%).The prevalences of AA were 1.58%,0.19% and 0.03% in the firs t-,second-and third-degree relatives of the probands respectively,which were significantly higher than those in the controls(P

Objective To analyze the differential proteomics of ASMC stimulated by wild IL-13 and mutant IL-13 and to investigate the relations of protein profiles of ASMC to asthma and possible targets for the treatment of bronchial asthma.Methods The total proteins of ASMC stimulated by wild IL-13 and mutant IL-13 were separated by immobilized pH gradient(IPG)-based 2-DE and the differentially expressed protein spots were identified by matrix assisted laser desorption-time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE detected approximately(840?21)spots on wild IL-13 samples and(892?17)spots on mutant IL-13 samples(n=3)and(685?19)spots matched.Six significantly differential proteins were subjected to MALDI-TOF-MS analysis and three of them were identified as stathmin 1,Ribosomal protein p~0 and NADH dehydrogenase.Conclusions ASMCs stimulated by wild IL-13 and mutant IL-13 present different proteomic profiles that may shed some light on the mechanism for the asthma causing effect of wild IL-13 and mutant IL-13.

Objective To investigate the effect of sevoflurane postconditioning on oxidative stress responses during focal cerebral ischemia-reperfusion (I/R) in rats.Methods Twenty-four male Wistar rats,weighing 240-280 g,were randomly assigned into 3 groups:sham operation group (group S),focal cerebral I/R group (group I/R) and sevoflurane postconditioning group (group SP).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 350 mg/kg.Focal cerebral I/R was produced by middle cerebral artery occlusion.In group SP,3.9％ sevoflurane (1.5 MAC) was inhaled starting from 20 min before reperfusion until 10 min after reperfusion.While 100％ O2 and air were given instead of sevoflurane in groups I/R and S,respectively.Six rats chosen from each group at 24 h of reperfusion were sacrificed and brains were removed for determination of malondialdehyde (MDA),glutathione (GSH),superoxide dismutase (SOD),catalase (CAT),glutathione peroxidase (GSH-Px) and glutathione reductase (GR) levels and for microscopic examination.The cerebral infarct size was measured by TTC staining.Results Compared with group S,MDA level and cerebral infarct size were significantly increased in groups I/R and SP,and GSH,SOD,CAT,GSH-Px and GR levels were decreased in group I/R,and GSH-Px level was decreased in group SP (P ＜ 0.05).Compared with group I/R,cerebral infarct size and MDA level were decreased,and GSH,SOD,CAT,GSH-Px and GR levels were decreased in group SP (P ＜ 0.05).The pathological changes were significantly attenuated in group SP compared with group I/R.Conclusion The mechanism by which sevoflurane postconditioning mitigates focal cerebral I/R injury in rats is related to enhanced antioxidase activity and inhibition of oxidative stress responses.

Objective A new method for detecting K-ras mutations based liquid chip was used to evaluate K-ras mutations associated with non-small cell lung cancer (NSCLC) patients,to direct the personalized treatment and prognosis evaluation.Methods Take the diagnosis technology research methods,the sensitivity and repeatability of the liquid chip K-ras gene mutation detection method were assessed.A total of 100 NSCLC patients from Nov 2011 to Feb 2012 in Shanghai Chest hospital were included in this study,the fresh tumor tissues were collected for DNA extraction.The 2nd exon 12 and 13 codons,containing 8 K-ras mutations occuring in high frequency were amplified by polymerase chain reaction (PCR),followed by ligation of the PCR products to a series of special probes using ligase detection reaction (LDR),then the PCR-LDR products were analyzed by liquid chip platform.Direct sequencing was applied to compare with the detection results.Results The sensitivity of liquid chip technology detection was 10％-20％,higher than the traditional sequencing method by 1％.Average CV value was 4％-15％ and showed good repeatability.5 K-ras mutations in 100 patients (5％) were detected using multiplex PCR-LDR combined fluid chip methods,including 3 Glyl2Val and 2 Gly12Asp mutations in exon 2.The 5 K-ras mutations were verified accurately by direct sequencing.Conclusions The novel detection method of K-ras mutations based PCRLDR and fluid chip shows high throughput,high sensitivity,good repeatability and the results are reliable and accurate.This method can be used to accurately identified K-ras mutations for NSCLC patients prior to their targeted therapy with TKIs.

ObjectiveTo observe protective effects of agmatine (AGM) on inflammatory response and spleen immune function in mice with trauma.Methods Forty-eight adult male C57BL/6 mice were randomly divided into three groups (n= 16 each), including control group, model group (bilateral femoral fracture and removal of 35% of the total blood volume), and AGM group (trauma/hemorrhage & AGM 200 mg/kg). Eight mice in each group were sacrificed at 3 hours and 24 hours, respectively, after modeling, and blood samples and tissue homogenate of spleen and liver were collected. The contents of tumor necrosis factor-α (TNF-α), interleukins (IL-6, IL-1β) in serum and liver tissue were determined with enzyme linked immunosorbent assay (ELISA). Serum aspartate transaminase (AST), alanine aminotransferase (ALT) and lactic dehydrogenase (LDH) were determined with automatic biochemistry analyzer. Spleen proliferation response stimulated with concanavalin A (ConA) was evaluated with methyl thiazolyl tetrazolium colourimetry (MTT).γ-interferon (IFN-γ) and IL-2 releases were determined with ELISA.Results Compared with control group, 3 hours after trauma/hemorrhage, the levels of serum TNF-α, IL-6, and IL-1β in model group were significantly elevated [TNF-α (ng/L): 145.38±31.50 vs. 23.06±11.14, IL-6 (ng/L): 496.94±50.76 vs. 47.13±17.47, IL-1β (ng/L): 321.31±43.02 vs. 29.25±16.24,allP< 0.01]. It was found that AGM treatment could alleviate the increase in serum pro-inflammatory mediators induced by trauma/hemorrhage, such as TNF-α (ng/L:111.56±25.47 vs. 145.38±31.50), IL-6 (ng/L: 412.56±44.33 vs. 496.94±50.76), IL-1β (ng/L: 273.38±45.25 vs. 321.31±43.02,P< 0.05 orP< 0.01). Twenty-four hours after trauma/hemorrhage, serum pro-inflammatory mediators were recovered to the levels in control group. There was no significant difference in TNF-α and IL-6 levels at 3 hours after trauma/hemorrhage among groups. Compared with control group, the expressions of liver TNF-α and IL-6 in model group were increased at 24 hours following trauma [TNF-α (ng/mg): 32.93±4.90 vs. 26.58±2.33, IL-6 (ng/mg): 11.20±1.66 vs. 8.38±0.89,bothP< 0.01]. However, AGM inhibited the level of TNF-α (ng/mg:28.92±3.16 vs. 32.93±4.90) and IL-6 (ng/mg: 9.03±1.28 vs. 11.20±1.66) in the liver as induced by trauma/hemorrhage (P< 0.05 andP< 0.01). At 24 hours after modeling, model group and AGM group had distinctly higher serum AST, ALT, LDH levels than those of control group [AST (U/L): 405.9±31.2, 245.7±22.1 vs. 128.2±15.9; ALT (U/L): 92.1±6.3, 51.6±5.0 vs. 30.1±3.2; LDH (U/L): 606.7±36.3, 478.7±25.3 vs. 384.0±16.6, allP< 0.01]. Nevertheless,the increase in serum AST, ALT and LDH was alleviated in AGM group (allP< 0.01). Meantime, trauma/hemorrhage produced a noticeable depression of proliferation of splenic cells and IFN-γ and IL-2 release stimulated with ConA compared with control group [proliferation rate: (40.97±4.13)% vs. (89.99±7.76)%, IFN-γ(ng/L): 91.6±12.3 vs. 353.2±21.5,IL-2 (ng/L): 53.4±6.4 vs. 91.0±12.2,allP< 0.01]. In contrast, AGM notably restored the capacity of proliferation response of splenic cells [proliferation rate: (74.86±5.75)% vs. (40.97±4.13)%, P< 0.01],enhanced the release of IFN-γ and IL-2 stimulated with ConA [IFN-γ (ng/L): 327.8±23.6 vs. 91.6±12.3, IL-2 (ng/L): 74.8±10.4 vs. 53.4±6.4, bothP< 0.01].Conclusion AGM can dramatically alleviate spleen immunosuppression, excessive inflammation and organ damage induced by trauma/hemorrhage.