The construction of the model curricula is the important branch of teaching quality engineering. The key to the development of the model curricula lies in the reform of the teaching model. As required to the national standards of the model curricula and to the purpose of innovative educational philosophy,with the years of continuous exploration and practice,the clinical immunology examination curriculum has kept developing from the teaching staff,teaching condition,content of course,teaching method and means etc.,which has brought about satisfactory teaching effects. Thus it is forming the model curricula with special features.

Objective To investigate the effects of macrophage migration inhibitory factor (MIF) overexpression on the expression of interleukin-8 (IL-8),martix metalloproteinase-9 (MMP-9) and invasion of human cervical cancer SiHa cells.Methods Chemical synthesis MIF eDNA gene,designed primer sequence including XhoI and BamHI enzyme sites,MIF gene was amplified by polymerase chain reaction (PCR),constructed eukaryotic expression vector pEGFP-N1/MIF and transfected into SiHa cells using Lipofectamine and won over-expression of MIF.The expression of MIF in supernatant fluid was detected by ELISA,the expression of MIF,IL-8,MMP-9 in both mRNA and protein levels were detected by real-time fluorescence quantitative-PCR and immunocytochemistry respectively.The effect of over-expressed MIF on migration was detected by Boyden small chamber.Results The expression of protein in supernatant fluid transfected with pEGFP-N1/MIF was significantly increased (Fgroup =8267.564,P ＜ 0.01),the expression of MIF,IL-8,MMP-9 in both mRNA and protein in SiHa cells transfected with pEGFP-N1/MIF were significantly increased (F values were 7019.619,2148.094,3303.540,1565.114,2807.300,523.466,P ＜ 0.01),and there was a positive correlation among MIF,IL-8,MMP-9 expression in both mRNA and protein (r values were 0.865,0.895,0.934,0.908,P ＜ 0.01).Invasion ability in SiHa cells transfected with pEGFP-N1/MIF was obviously increased (F=3430.898,P＜ 0.01).Conclusion The over-expression MIF gene in SiHa cells can promote cervical cancer cell invasion and metastasis of ability,which could be associated with the upregulation of IL-8 and MMP-9 expression.

Objective To elucidate whether down-regulation of homo sapiens eukaryotic translation elongation factor 1 alpha 2 (EEF1A2) expression induces apoptosis in pancreatic cancer cells and its possible mechanisms. Methods Two siRNAs targeting human EEF1A2 were synthesized and the siRNA/liposome complexes were transfected into the pancreatic cancer cell line BxPC-3. RTPCR and Western blot were used to analyze the change of EEF1A2 expression and the apoptosis rate of BxPC-3 cells was studied using Annexin-V/PI assay. To identify the mechanisms involved, the apoptosis associated proteins such as caspase-3, caspase-8, caspase-9, PARP, cytochrome C and Bid were detected by Western blotting. Results Both EEF1A2-targeting siRNAs reduced the EEF1A2expression, and the No. 2 siRNA inhibited EEF1A2 expression to less than 25 % in mRNA and protein levels. Down-regulation of EEF1A2 expression in BxPC-3 cells enhanced cell apoptosis (15.28% ±3.65%) at a greater level than negative siRNA-expressing cells (10. 11% ± 3. 05%) or mock cells (9.41 % ±4.14 %). Furthermore, reduction of EEF1A2 activated the pro-caspase-8, pro-caspase-3,pro-caspase-9,PARP and Bid to their active forms, and increased the expression of cytochrome C.Conclusions These data suggest that EEF1A2 down-regulation could significantly induce apoptosis of pancreatic cancer cell line BxPC-3, which is likely mediated by the death receptor and mitochondrial apoptotic pathways.

Clinical practice is an important stage in clinical education and the future career of the graduates. Taking the students of Clinical Inspection Major as an example,this article analyzes the questions need to be paid attention to in the clinical practice and the importance of pre-post training in increasing the quality of clinical practice.

Chemical disinfectants are generally used for virus inactivation and environment disinfection in biosafety laboratory, and the efficacy and evaluation of the disinfection are critical to ensure the laboratory biosafety.However, there is a current lack of applied standard to evaluate the virucidal efficacy of chemical disinfectants in our country.In this paper, a European Union standard“Method and Requirements of Virucidal Quantitative Suspension Test Method for Chemical Disinfectants Used in Human Medicine” was analyzed and a standard transformation scheme has been proposed.It is suggested that the model viruses should be increased from 3 to 6, including the surrogate viruses to substitute highly pathogenic viruses, and that the method to remove the residual chemical disinfectant and the calculation of 95%confidence interval should be incorporated into the standard.The suggestion will improve the scientific and operational standards related to disinfection and sterilization in biosafety laboratory in China.

Objective To explore the effect and the possible mechanisms of silent homo sapiens eukaryotic translation elongation factor 1 alpha 2(EEF1A2)gene on the growth of pancreatic cancer cell in vivo.Methods The pancreatic cancer xenograft models in mice were established.The mice were equally divided into control group,negative control group and EEF1A2 group,which were injected with PBS,negative control siRNA and EEF1A2 siRNA into xenograft tumors respectively.The size and weight of tumors in each group were measured.The expression of EEF1A2 and PCNA in tumor tissue of each group was detected by immunohistochemistry.The cell apoptosis rate in tumor tissue of each group was determined by TUNEL.Results In xenograft nude mice models,since the 17th day of injection,the growth of tumor size in EEF1A2 group was obviously slower than that of negative control group and control group(all P＜0.05).By the end of the treatment,the tumors were cut off and weighted.The weight of tumors in EEF1A2 group(0.27g± 0.06g)were significantly lower than those of control group and negative control group(0.39g± 0.08g and 0.43g± 0.07g,P＜0.05).EEF1A2 mostly expressed in cytoplasm of pancreatic cancer cell.In negative control group and control group,the positive cells distributed densely and the positive rate was(72.58 ± 25.47)％ and (76.75±23.19)％ respectively.The distribution of positive cells in EEF1A2 group was scattered and the positive rate was(34.78±21.36)％,the difference was statisically significant(P＜0.01).The expression of PCNA at protein level in EEF1A2 group was significantly lower those that of control group and negative control group(P＜ 0.01).The result of TUNEL test indicated that the cell apoptosis rate in EEF1A2 group was higher than those of control group and negative control group (P＜0.01).Conclusions The EEF1A2 gene can be effectively silented in vivo,which significantly inhibits the growth of pancreatic cancer cell.It may be related with inhibition of cell proliferation and promotion cell apoptosis.

Objective To investigate the effects of the expressions of thymidine kinase 1 (TK1) and Ki67 alone or their combination on the recurrence and metastasis of breast cancer.Methods Sixty-five samples were selected from the Second Affiliated Hospital of Guangzhou Medical University from March 2005 to June 2007,which were resected by surgical operation and confirmed as breast carcinoma by pathology.They were individed into two groups including 37 cases with recurrence or metastasis in 5 years (group A),28 cases without recurrence or metastasis in 5 years (group B).The expressions of TK1 and Ki67 in the two groups were detected by immunohistochemical staining assay.Then,Kaplan-Meier assay was used to describe survival curve.Results The positive expression rate of TK1 in group A was 91.8％,which was dramatically higher than that in group B 67.8％ (x2 =6.116,P =0.013).The positive expression rates of Ki67 in group A and B were 78.4％ and 42.9％ (x2 =8.635,P =0.003).The positive expression rates of TK1 combined with Ki67 in group A and B were 67.6％ and 39.3％ (x2 =5.159,P =0.023).Moreover,disease free survival of patients with positive expression of TK1 combined with Ki67 decreased significantly,compared with patients with positive expression of TK1 or Ki67 alone (x2 =6.137,P =0.046).Conclusion Positive expression of TK1 combined with Ki67 is the high risk factor of the reccurence or metastasis of breast carcinoma,and indicates poorer prognosis compared with positive expression alone.

Objective To develop a sandwich enzyme-linked immunosorbent assay ( ELISA) for the determination of a humanized antibody MIL50.Methods RiVax, a mutant of RTA (a chain of ricin) expressed in E.coli, was used as the coating protein.Horseradish peroxidase (HRP) labeled IgG of goats against humans was used to determine MIL 50 captured by RiVax coated on the plate.Results Compared with ricin, RiVax could bind well with MIL50,indicating it could be used as a coating protein to determine MIL50 instead of holotoxin.Using this method, the detective limit of MIL50 in PBST (PBS containing 0.1%Tween 20) could be as low as 0.030 mg/L.The absorbance value of ELISA for MIL50 in the ser-um and homogenate of the liver , spleen, lung, kidney, muscle of rats was lower than that in PBST .Conclusion The sandwich ELISA is a sensitive method for the analysis of distribution of MIL 50 in rat tissues.

Objective To investigate the prevalence of anxiety and depression in elderly patients with metabolic syndrome (MS) and its related factors.Methods A total of 672 subjects aged 60 or over undergoing health check-up in two Nanjing community health service centers from November 2015 to October 2016 were enrolled in the study.The basic information and the history of hyperlipidemia,hypertension and diabetes were collected by questionnaire survey;the results of physical examination and biochemical testing were documented.The prevalence of anxiety and depression were investigated by the Hospital Anxiety and Depression Scale (HADS).According to MS diagnostic criteria,the subjects were divided into MS group (n=181) and non-MS group (n=491).The HADS scores of two groups were compared and the influencing factors related to anxiety and depression were analyzed.Results The levels of systolic blood pressure (SBP),diastolic blood pressure (DBP),body mass index(BMI),levels of fasting blood glucose (FBG),triglyceride (TG),low density lipoprotein cholesterol (LDL-C) were higher and high density lipoprotein cholesterol (HDL-C) was lower in MS group than those in non-MS group (all P<0.05).The prevalence rates of anxiety and depression in MS group (30.9% and 34.8%) were significantly higher than those in non-MS group (20.2% vs.25.1%,χ2=8.655,6.288,P=0.003,0.012).Multivariate logistic regression analysis showed that obesity (BMI≥28 kg/m2),high FBG (≥7.0 mmol/L),hypertension [blood Pressure≥140/90 mmHg (1 mmHg=0.133 kPa)] were the independent risk factors for anxiety in MS patients (OR=3.987,2.827,2.375,respectively,all P<0.05);obesity (BMI≥28 kg/m2),high FBG(≥7.0 mmol/L),smoke,high TC (≥5.2 mmol/L),hypertension (≥140/90 mmHg) were the independent risk factors for depression in MS patients (OR=7.718,3.233,2.071,1.932,1.910,respectively,all P<0.05).Conclusion Elderly patients with metabolic syndrome are prone to anxiety and depression,and obesity,high FBG,hypertension and other factors are the risk factors for anxiety and depression.

Objective To investigate the expression of oxidative stress related factors,such as,superoxide dismutase (SOD),heme oxygenase-1 (HO-1),malondialdehyde (MDA) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) in patients with idiopathic membranous nephropathy (IMN) and explore the possible role of oxidative stress in the pathogenesis of IMN.Methods Serum,urine and renal tissue of 40 patients with IMN,20 patients with secondary membranous nephropathy (SMN),and 20 patients with minimal change nephropathy (MCD) were collected from the nephrology department of the First Affiliated Hospital of Zhengzhou University.All of them were diagnosed by renal biopsy from January 2016 to June 2016.Serum,urine and renal tissue of 20 healthy persons were collected as the normal control (NC).The expressions of SOD,HO-1,and 8-OHdG in renal tissues were detected by immunohistochemistry.The levels of SOD,HO-1,MDA and 8-OHdG in serum and urine specimens were detected by ELISA.The relationship between oxidative stress related factors and proteinuria in IMN was investigated as well.Results The expressions of SOD,HO-1 and 8-OHdG were significantly higher than those in MCD group and NC group in renal tissue;the expression of SOD in IMN group was significantly higher than that in MCD group and NC group in serum (all P ＜ 0.01).The expressions of HO-1,MDA and 8-OHdG in the serum and urine of the IMN group were significantly higher than those of the MCD group and the NC group (all P ＜ 0.05).There was a significant positive correlation between the urine expression of 8-OHdG and MDA and proteinuria in the IMN group.Conclusions Oxidative stress-related factors in IMN patients are elevated,which is closely related to clinical manifestations.In that sense,it is confirmed that oxidative stress is involved in the pathogenesis of IMN.