Objective To evaluate the potential association of CD4+CD25+ T cells with alopecia areata.Methods Totally,this study enrolled 23 patients with progressive alopecia areata,25 patients with stable alopecia areata,and 25 healthy controls.Peripheral blood was isolated from these subjects followed by isolation of CD4+ CD25+ regulatory T cells,which were then cuhured with the presence of anti-CD3 and-CD28 monoclonal antibodies for four days.Subsequently,enzyme-linked immunosorbent assay was performed to measure the levels of interleukin (IL)-10 and transforming growth factor (TGF)-β1 in the culture supematant of these T cells.Results The levels of IL-10 and TGF-β1 were (31.68 ± 6.78) pg/ml and (32.29 ± 6.8) pg/ml respectively in the culture supernatant of CD4+CD25+ regulatory T cells from patients with progressive alopecia areata,significantly lower than those from the healthy controls ((57.34 ± 14.15) pg/ml and (57.43 ± 15.16) pg/ml,both P ＜ 0.05) and patients with stable alopecia areata ((52.56 ± 13.02) pg/ml and (61.75 ± 14.10) pg/ml,both P ＜ 0.05).However,no significant difference was observed in the supernatant levels of IL-10 or TGF-β1 between the healthy controls and patients with stable alopecia areata.Conclusions The secretion of IL-10 and TGF-β1 by CD4+CD25+ T cells is decreased in patients with progressive alopecia areata,which may contribute to the pathogenesis of alopecia areata.

OBJECTIVE To set up an intranasal ovalbumin-induced animal model of allergic rhinitis(AR) accompanied with olfactory dysfunction in mice. By observing the olfactory pathway in mice using manganese-enhanced magnetic resonance imaging (MEMRI) and the relatively morphologic structural and immunological changes in olfactory epithelium, the influence of AR on olfactory receptor neurons(ORNs) was studied.METHODS Forty SD mice were randomly divided into two groups, the research group(n=30) and the control group(n=10). The research group was intraperitoneally injected and intranasal application of ovalbumin to establish an AR mice model. The olfactory function of the mice was evaluated by buried food test(BFT). ELISA was performed to measure the level of IgE in serum. MEMRI images were acquired with a 7.0 T micro-MR scanner. HE staining and immunohistochemistry were used to observe the tissues morphology change of olfactory mucosa and OMP expression.RESULTS The olfactory function evaluation of the AR mice model indicated that the incidence of olfactory dysfunction in AR mice was 40.0%. The AR mice with olfactory dysfunction had no signal enhancement in MEMRI. The olfactory epithelium became thinner, layer numbers of ORNs were decreased with disorder arrangement and the OMP expression was decreased in AR mice with olfactory dysfunction compared with that in AR mice without olfactory dysfunction(P=0.018) and the control group(P=0.0141).CONCLUSION An animal model of AR accompanied with olfactory dysfunction in mice was successfully established. The influence of AR on ORNs and thus cause the change of the olfactory pathway is one of the major pathogenesis of olfactory dysfunction in AR.

ObjectiveTo study the application of three-dimensional simulated surgical technique in precise hepatectomy. MethodsFrom July 2009 to February 2010, 16 patients with primary liver cancer underwent preoperative simulated imaging and three-dimensional simulation of liver resection.The 3D extent of simulated hepatectomy and actual hepatectomy was compared and analyzed. ResultsThe shape and the extent of the liver resected were very similar in the simulated and the actual hepatectomies. The mean differences in the length, breadth and depth of the remnant livers were 0. 6118 cm,0. 4490 cm and 0. 3199 cm, respectively. ConclusionsSimulation hepatectomy could predict the extent of the actual liver resection, and provided accurate guidance and preoperative planning for precise hepatectomy.

BACKGROUND: Biological characteristics of porcine cardiovascular system are similar to that of human. The metabolism, immune system, mechanism of disease have 99% homology between pig and human. OBJECTIVE: To develop ventricular aneurysm animal models by plugging left anterior descending branch. DESIGN, TIME AND SETTING: The animal observational study was performed at the Laboratory of Functional Material and Animal Experimental Center, Tongji Hospital Affiliated to Tongji University from December 2005 to July 2007. MATERIALS: A total of 13 pigs, of both genders, weighing 30-40 kg, were used in this study. METHODS: After abdominal cavity and intravenous anesthesia, balloon with "WYW" stent was put into 13 pig left anterior descending coronary artery following No. 7 artery sheathing canal was implanted. The balloon was dilated and the stent was positioned into the distal point of the first diagonal branch to obstruct the artery under digital subtraction angiography. During and after the operation, electrocardiogram was monitored and recorded to maintain stable vital sign. MAIN OUTCOME MEASURES: Electrocardiogram, serum myocardial enzymes, myocardial radionuciide imaging, echocardiography and angiography and pathological changes were observed. RESULTS: One rat died of anesthetic accident before surgery. Six rats died from ventricular fibrillation during plugging, and the other 6 rats were survival. 4 weeks following surgery, coronary angiography showed blood flow at distal end was blocked 100%. Left ventricle angiography demonstrated that wall motion at apex of the heart and the left ventricle disappeared. Before embolism, electrocardio-monitoring displayed electrocardiogram was normal. After plugging, ST segment was raised continuously; amplitude of R wave was decreased; T wave was erected; ST-T fusion wave appeared; precordial leads were obvious. ST segment was reduced to basic level 2 week later, and pathological Q wave appeared 4 weeks later. Serum treponin was increased at 12 hears following surgery compared with before surgery (P < 0.01). Radioactive nuclide myocardial imaging revealed that cardiac apex and left ventdcle anterior wall showed radioactive nuclide perfusion filling defect, thin ventricular wall and ventricular aneurysm. Cardiac ultrasonic showed six pigs suffered from weaken segmentation contraction motor of local ventricular wall, especially at anterior wall of the left ventricle and cardiac apex. After hematoxylin-eesin staining, optical microscope exhibited that cardiac muscle fiber disappeared, which was replaced by collagen fiber, with the presence of some capillary at the infarct of cardiac apex. Pyknosis and lysis of residual cardiac muscle fiber were visible in infarct region of anterior wall of the right ventricle. Many inflammatory cells and capillary infiltration were detected besides collagen fiber filling. CONCLUSION: The new approach of "WYW" intervention embolization is able to develop the ventricular aneurysm animal model, which is similar to clinical pathophysiological variation. The outcome is precise and reliable.

Objective To observe the targeted function of a mesothelin antibody modified nanoprobe in human pancreatic cancer BxPC3 cell.Methods The Fe3O4@SiO2 nanoprobe was prepared by St(o)ber method,and then quantum dots (CdTe) and mesothelin antibody was crosslinked to obtain the properties of targeting and fluorescent.Fluorescent nano Fe3O4@SiO2 probes and BxPC3 cells were incubated in vitro for 30 min.Its targeting performance was tested by the CCD imaging system and magnetic separation technology.HepG-2 and K562 cells with low expression of mesothelin were selected as reference cells.Results This preparation method of nanoprobe could produce a uniform and narrow distribution particle with particle size mainly ranging from 120 to 140 nm.The cell adsorption experiments showed that the adsorption efficiency of BxPC3,HepG-2 and K562 by nanoprobe without crosslinking antibody were less than 20％,as a non-specific adsorption; and the adsorption efficiency of BxPC3,HepG-2 and K562 by crosslinking mesothelin antibody nanoprobe were (53.9 ± 1.8) ％,(8.0 ± 2.1) ％ and (8.9 ± 2.3) ％ respectively,and the adsorption capacity with BxPC3 was significantly increased.Conclusions The nanoprobe modified by mesothelin antibody can effectively recognize BxPC3 cells which highly expressing mesothelin.

Objective To explore the relationship between blood lipid level and primary open angle glaucoma(POAG)and intraocular pressure(IOP).Methods A retrospective case-control study was designed.80 POAG patients from EYE & ENT hospital of FUDAN university were collected as a case group from January 2015 to May 2016.Meanwhile 80 age and gender matched healthy people were collected as a control group.The differences of gender in men subgroup (POAG:44,Control:39) and female subgroup (POAG:36,Control:41) showed no statistic significance.Serum total cholesterol(TC),total triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),apolipoprotein A(Apo A) and apolipoprotein B(Apo B) were measured using Roche automatic biochemical analyzer.The analysis of the above mentioned lipid levels between POAG group and control group was conducted by independent student′s T test and multivariate logistic regression.The relationship between lipid level and IOP was analysied using Pearson correlation test.Results The TG level was (1.40±0.71)mmol/L in the POAG group,(1.00±0.37)mmol/L in the control group.The difference had statistic significance (t=4.467,P<0.001).The HDL-C level in the POAG group was (1.22±0.36)mmol/L,(1.38±0.25)mmol/L in the control group.The difference also had statistic significance (t=-3.228,P=0.002).Comparison between the men subgroup and female subgroup found that the TG level in the POAG group of men was (1.47±0.71)mmol/L,(1.01±0.36) mmol/L in the control group and the HDL-C level were (1.14±0.31) mmol/L,(1.32±0.20)mmol/L in two groups.Both the level of TG and HDL-C had statistic significance in men (t=3.717,P<0.001;t=-3.240,P=0.002).The TC,TG level in the POAG group and control group of women respectively were (4.59±0.81)mmol/L,(1.33±0.72)mmol/L and (4.21±0.64)mmol/L,(0.99±0.38)mmol/L.Both blood level were significantly higher than the control group in women (t=2.278,P=0.026;t=2.503,P=0.016).Logistic regression analysis showed that the TG level correlated with POAG (P=0.004,OR=2.831,95% CI=1.392-5.759)and both POAG in the men and women (P=0.025,OR=3.223,95% CI=1.156-8.989;P=0.045,OR=2.554,95% CI=1.020-6.395).Pearson test showed that the TG level of POAG patients was positively associated with IOP in spite of gender(P<0.05).Conclusion The peripheral blood level of TG was significantly increased in the POAG patients and significantly correlated with IOP,which indicated that the increased peripheral blood level of TG may be the independent risk factor for the POAG..

Objective A large sample of multi-factor Logistic analysis of clinical biochemical indicators from elderly cataract patients were done to find the characteristics of possible changes in potential factors for cataract-related indicators and explore the pathogenesis of cataract from the perspective of laboratory diagnosis.Methods A retrospective case control study was conducted .Cataract patients from Eye-Ear Nose Throat Hospital of Fudan University were collected during June 2011 to July 2015 and routinely tested for 14 clinical biochemistry indicators including ALB , GLU, TP, GLB, BUN, URCA, Cr, LDH, ALP, Ca, Na, K, Cl, and P.The mean age of the 3 527 cataract patients (1 804 males, 1 723 females) was (64.47 ±11.29) years.Those data were reserved by LIS (Laboratory Information System) system and were retrieved for large data analysis . The control group was filtered from hospitalized patients besides cataract patients during January 2014 to July 2015.The mean age of the 3 333 control subjects ( 1 770 males, 1 563 females ) was ( 64.04 ±9.03 ) years.Biochemical indicators between cataract group and control group were analyzed by independent student′s t-test and Logistic regression .Meanwhile , the objects were divided into 3 groups according to the age (40-59 years,60-79 years,≥80 years) for independent student′s t-test with the control group respectively .Results There are no statistical difference in age ( t=1.663,P=0.096) and gender ( t=2.63,P=0.105 0) between cataract group and control group ( P>0.05).The results of Logistic regression between cataract group and control group were as follow : ALB (OR=1.053,95%CI=1.019-1.088),BUN(OR=1.113,95% CI=1.076-1.152),Cr(OR=1.007, 95%CI=1.003-1.010),GLB(OR=1.049,95% CI=1.018 -1.081),GLU(OR=1.175,95% CI=1.139-1.211),Cl(OR=1.059,1.032 -1.088),Na(OR=1.180,95% CI =1.150 -1.212).Those might be enrolled as risk factors for cataract (P<0.05).The level of ALB, GLU, Na and Cl in cataract group according to different age groups were significantly higher ( t=8.780,t=4.670, t=9.695, t=6.415, P<0.05) than that of control in 40-59 age group;the level of ALB, BUN, Cr, GLU, Na and Cl in cataract group was significantly higher ( t=4.974, t=9.414, t=4.123, t=6.906, t=14.741, t=9.212, P<0.05) than control in 60 -79 age group; the level of BUN, Cr, GLU, Na and Cl in cataract group was significantly higher (t=6.077, t=4.906, t=2.626, t=5.459, t=3.424, P<0.05) than control in 80-age group.Conclusions The level of ALB, BUN, GLB, Cr, GLU, Na and Cl in peripheral blood of cataract patients was significantly increased which suggested that those change might associated with the pathogenesis of cataract .

Objective To study relationships between myocardial injury and the levels of serum complement C3, C4 and C5b-9 in patients with acute coronary syndrome (ACS). Methods A retrospectively analysis was conducted. 170 ACS patients [including 110 cases of ST-segment elevation myocardial infarction (STEMI) and 60 cases of non-ST-segment elevation acute coronary syndrome (NSTE-ACS)] with ischemic chest pain or chest discomfort onset within the prior 12 hours admitted to the cardiology department of Tianjin Union Medicine Center from January 2014 to July 2016 were enrolled. Thirty-six healthy cases were enrolled as control during the same time. The levels of serum complement C3, C4 and C5b-9 on 1, 3 and 7 days after admission and myocardial function indicators were analyzed. Major adverse cardiovascular events (MACE) and readmission rate were analyzed after 1 year follow-up. The correlation between serum complement levels and myocardial function indicators was analyzed by Pearson correlation analysis. Results ① The levels of serum C3, C4 and C5b-9 on the first day in NSTE-ACS group and STEMI group were significantly higher than control group [C3 (g/L): 1.04±0.33, 1.26±0.35 vs. 0.39±0.21, C4 (g/L): 0.31±0.14, 0.33±0.10 vs. 0.19±0.07, C5b-9 (g/L): 575.46±197.26, 659.26±160.77 vs. 501.40±141.51, all P < 0.05]. There were no changes of serum C3, C4 in NSTE-ACS group, but C5b-9 decreased after a peak (g/L: 700.63±218.42) at 3 days. Serum complements in STEMI group reached peak on the third day [C3 (g/L): 1.37±0.33, C4 (g/L): 0.42±0.12, C5b-9 (g/L): 754.72±136.22]. The levels of serum C4 and C5b-9 in STEMI group were higher than NSTE-ACS group on the third and seventh day. ② The levels of troponin T (TnT), creatine kinase-MB (CK-MB), solution intercellular adhesion molecule-1 (sICAM-1), global registry of acute coronary events (GRACE) scores and percutaneous coronary intervention (PCI) numbers in STEMI group were significantly higher than those in the NSTE-ACS group, which were as opposite as left ventricular ejection fraction (LVEF). However, there were no significant differences in levels of serum N-terminal pro-brain nitric peptide (NT-proBNP), Fibrinogen (Fib), readmission rate and incidence of MACE between STEMI and NSTE-ACS groups. ③ According to GRACE, patients with ACS were divided into low risk group (≤ 108 scores, 26 cases), intermediate risk group (109-140 scores, 61 cases) and highest group (> 140 scores, 83 cases). TnT and sICAM-1 in intermediate risk group were significantly increased as compared with low risk group. Levels of TnT, sICAM-1, C3, C4 and C5b-9 in the highest group were significantly higher than the low and intermediate risk groups, however the lowest LVEF was found in the highest group. ④ It was shown by Pearson correlation analyses that levels of serum C3, C4, C5b-9 were positively correlated with TnT (r value was 0.481, 0.367, 0.292, respectively, all P <0.01), sICAM-1 (r value was 0.298, 0.249, 0.365, respectively, all P < 0.01), but negatively correlated with LVEF (r value was -0.384, -0.260, -0.200, respectively, all P < 0.01). In addition sICAM-1 positively correlated with TnT (r = 0.536, P = 0.000), but negatively correlated with LVEF (r = -0.341, P = 0.001). Conclusions Serum complements activation was found in the acute phase of ACS patients. Serum complement C3, C4 and C5b-9 are involved in the process of myocardial injury, and may reflect severity of myocardial injury and cardiac dysfunction.

Objective To analyze the effect of edaravone combined with ginkgo diterpene lactone in treatment of acute cerebral infarction (ACI) and discuss the better clinical treatment to ACI .Methods 100 cases of ACI and who accepted treat-ments in our hospital were selected as the research objects in January 2014 to January 2015 .50 patients as observation group were given the treatment of edaravone and ginkgo diterpene lactone at the same time ,while other 50 patients as control group were merely given the treatment of ginkgo diterpene lactone .At the same time ,both groups were given treatment of conven-tional therapy .The total neurological function ,effective rate ,adverse reactions ,and the treatment satisfaction of both groups before and after the treatment were observed .Results There were no significant different of NIHSS score of two groups before treatment (P>0 .05) .After treatment ,NIHSS score of the observation group was significantly lower than the control group (P<0 .05) .The total effective rate of observation group was 90％ ,which was significantly higher than the control group (66％ ,P<0 .05) .The main adverse reactions were rash ,nausea ,dizziness of two groups ,and there were no significant differ-ent of adverse reactions rate of two groups through the treatment (P>0 .05) .The treatment of patients with satisfaction rate of observation group was 94％ ,which was significantly higher than control group (84％ , P< 0.05) .Conclusion Edaravone combined with ginkgo diterpene lactone had a good effect on improving nerve function of ACI patients ,which could improve the treatment of patients with satisfaction ,high security ,and could be recommended in clinical .

The expression vector of SmIg scFv fragment was constructed in patient with B cell chronic lymphocyte leukemia (B-CLL) and expressed in E. coli to obtain scFv fragment, and the effect of the protein on the proliferation of stimulated peripheral blood mononuclear cells (PBMC) was investigated in vitro. Two pairs of primers were designed, and variable region genes of light chain and heavy chain were amplified by PCR respectively from the pGEM-T vectors previously constructed in our laboratory which containing light chain gene or Fd fragment of heavy chain gene. The PCR product was digested, purified and inserted into pHEN2 vector to construct the soluble expression vector pHEN2-scFv. After the induction by IPTG, the scFv protein was identified by SDS-PAGE electrophoresis and purified by Ni-NTA-Chromatography. MTT was used to determine the effect of purified protein on the proliferation of stimulated PBMC in vitro. Plasmid PCR and restriction enzyme digestion of pHEN2-scFv revealed the pHEN2-scFv vector was constructed successfully. Id-scFv protein was expressed in positive clone after induced by IPTG. SDS-PAGE analysis showed that the relative molecular weight of fusion protein was about 30 kD (1 kD= 0.9921 ku), which was consistent with the theoretically predicted value. Proliferation of PBMC could be induced by purified Id-scFv. It was suggested that the expression vector of SmIg scFv fragment was constructed successfully, and scFv protein was expressed and secreted from E. coli, which could induce proliferation of PBMC. This may lay an experimental foundation for further research of Id-HSP complex vaccine for B-CLL.