Functional experiment plays an important role in basic theory and clinical practice. For training high-quality medical talents in medical colleges,many improvements have been made in the functional experiment teaching and certain effects have been obtained. But a comprehensive evalu-ation system was still needed. Clinical medical students of 2010 grade were taken as the appraisal objects. Assessment content,methods and standards were designed according to the basic requirements of functional experiment. Students were evaluated by ordinary performance,test papers and experiment competition. Practice showed that the system was of significant importance in improving the experi-mental teaching quality and developing students' practical ability and comprehensive quality.

Objective To use HSV 1 tk (herpes simplex virus type Ⅰthymidine kinase)/GCV (ganciclovir) in androgen independent prostate cancer cells(C4 2,PC3) in vitro in order to provide useful basis for clinical use. Methods HSV 1 tk gene was ligated to a pN 2A retroviral vector. Recombinant DNA molecules being introduced into a packaging cell line PA317,the high titer virus producer cells (VPC) were screened.The integration and expression of HSV 1 tk gene in VPC was observed by PCR and RT PCR. VPC was co cultured with these cancer cells in the light of 1∶1,1∶2,1∶4,1∶8.Cell viability (cytotoxicity) was assessed by SRB (sulforhodamine B protein dye binding)after the first day,the third day,the fifth and the seventh day. Results The highest titer VPC producing HSV 1 tk gene was isolated. Retrovirus mediated HSV 1 tk gene therapy was effective and active against such prostate cancer cells.The best one was co culture of VPC and cancer cells at 1∶1 and the fifth day followed by GCV. Compared with C4 2, PC3 decreased remarkably.The activation of apoptosis and other ways failed to be found. Conclusions Retrovirus mediated HSV 1 tk gene therapy in Vitro directly killed the tumor cells by cytolytic activity.

Objective] To explore the nursing method and effect of traditional Chinese medicine wet compress for the treatment of the disease of the blood vessel. [Method]Selects the hospital March 2013 to January 2015 ,a total of 96 patients with confirmed thrombotic occlusion vasculitis with the first period and the second period of patients, who were randomly divided into treatment group and control group, 48 cases in each, two groups at the same time, the routine nursing and the context of Mailuoning 30ml IV drip, treatment group based on the Tougu Tongmai Recipe wet applied external treatment, combined with TCM syndrome nursing. Two groups for 1 month for a course of treatment, observation of cold pain in the limbs, gap of claudication, soreness and numbness symptoms and treatment of total efficiency; readmission after followed up to observe the recurrence(deterioration), statistical analysis of parallel. [Results] The total effective rates of treatment group and control group were 95.8% and 68.75% respectively, the difference was statistically significant(P<0.05); the difference between the two groups was statistically significant(P<0.05); the difference between the two groups was statistically significant(P<0.05). [Conclusion] Compared with Mailuoning injection in the treatment of, Speranskia through vein wet compress external treatment nursing can effectively improve the thrombus occlusive vasculitis in patients with limb peripheral blood circulation, also can better reduce the recurrence rate, slow the progression of the disease, so as to achieve the purpose of disease treatment.

Background The parameter of corneal diameter in myopic eyes is widely used in clinic,but there are different points of view about the correlation of corneal horizontal diameter with other parameters of the myopic eye.Objective This study was to investigate the relevance of the other parameters to the corneal horizontal diameter(CHD)of myopia.Methods A total of 310 cases(310 eyes)of myopic patients aged 6-50 years old who visited Affiliated First Hospital of Zhengzhou University were collected for the study.Measuring items included gender,age,myopia diopter and corneal topography.The relationships between the CHD of the right eyes and seven factors including age,gender,degree of myopia,corneal curvature (CC),corneal astigmatism (CS),corneal central thickness(CCT),and anterior chamber depth (ACD)were analyzed by empower stats software.Results The distribution range of CHD was from 10.8 mm to 13.5 mm,with the average value (11.7±3.8)mm.There were significant differences in the CC and ACD between male and female patients by t test(t =-1.574,P＜0.001 ;t=-1.145,P =0.034).Through the smoothing curve fitting,the threshold effect and single factor and multiple regression analysis,the CHD was negative linear relationship with CC (β =-0.085,P =0.011).The ACD positive linear relationship with CHD was found (β=0.722,P＜0.001).And the CHD was not correlated with the degree of myopia,CS,CCT and gender(β =0.000,0.084,-0.001,0.105;all at P＞0.05).There was different inflection point in the curve relationship between male patients and female patients.Conclusions The CHD is linear negatively correlated with CC and line positively associated with ACD in 6-50 years old myopic patients.There is no relationship between CHD and gender,spherical equivalent degree,CS,CCT.There is curvilinear relationship with inflection point between CHD and age.

[ ABSTRACT] AIM:To investigate the effect of microwave radiation at different intensities on the rat myocardium and its possible mechanism.METHODS:The rats were radiated by the intensity of 500, 1 000, 1 500 and 2 000 W/m2 with 2 450 MHz microwave for 6 min.The heart tissue was collected 6 h after microwave radiation.ATP and mitochondria complexⅣandⅤwere measured.The changes of the tissue structures were observed under transmission electron micro-scope.The apoptosis of the myocardial cells was detected by a cell analyzer.The protein level of cleaved caspase-3 was de-termined by Western blotting.RESULTS:The concentration of ATP and activity of mitochondria complexⅣandⅤsigni-ficantly decreased compared with control group in the cardiac tissues.The decreased number, morphological abnormalities such as dissolved cavitation, matrix and obvious tumefaction of mitochondria were observed under transmission electron mi-croscope.The microwave radiation induced the apoptosis of myocardial cells in the rats.The cell apoptotic rate and the pro-tein level of cleaved caspase-3 increased with increasing intensity of microwave radiation ( P<0.05 ) .CONCLUSION:Microwave radiation has obvious injury effect on the rat heart, which can cause cardiac energy metabolism dysregulation and cardiac myocyte apoptosis.

Aim To study the effect and molecular mechanism of pretreatment with COPP on hypoxia/reoxygenation injury of H9c2 myocytes.Methods H9c2 myocytes model of hypoxia/reoxygenation injury was established and H9c2 myocytes were given COPP pretreatment before hypoxia/reoxygenation.Treatment with Znpp and all-trans retinoic acid(ATRA)inhibited HO-1 and Nrf2-ARE respectively.The level of LDH and CK in cell supernatants were measured.HO-1mRNA expression was analyzed by RT-PCR.HO-1 and Nrf2 protein expressions were analyzed by Western blot.Results Compared with hypoxia/reoxygenation group,the level of LDH and CK in COPP pretreatment groups decreased significantly and the level of HO-1mRNA,HO-1 protein expression and Nrf2 protein expression in the nucleus significantly increased.Znpp abolished protective effect of COPP pretreatment.ATRA blocked the nuclear accumulation of Nrf2 and decreased HO-1 protein expression that COPP pretreatment induced.Conclusions COPP can induce HO-1 overexpression which has protective effect on hypoxia/reoxygenation injury of H9c2 myocytes.Its mechanism is related to Nrf2-ARE signaling pathway.

Objective To investigate the effects of small interference RNA (siRNA) targeting HOXA9 on the proliferation and apoptosis of human acute monocytic leukemia U937 cell line.Methods Effective and specific siRNA oligo targeting HOXA9 was designed and compounded.It was transfected transiently into U937 cells by cationic liposome.The cells was divided into three groups:experimental group(siRNA targeting HOXA9 was transfected by liposome),negative control group (negative siRNA was transfected by liposome) and cell control group (add equal cells and medium).The expression of HOXA9 mRNA and protein were detected by reverse transcription PCR and Western blot.The cell proliferation was assessed by MTT.The apoptosis of each group were measured by Annexin V-FITC.Results Aftcr transfected by siRNA targeting HOXA9,the relative mRNA expression levels of HOXA9 in the experimental group,negative control group and cell control group were (22.980±0.548) ％,(82.371±1.517) ％ and (84.637±2.252) ％,respectively (P ＜ 0.05),and the relative protein expression levels were (50.377±2.773).％,(105.500±3.900) ％ and (111.392±3.905) ％,respectively (P ＜ 0.05).The inhibitory rates of cell proliferation and the apoptosis rates of the experimental group were significantly increased.The inhibitory rates of cell proliferation of 24 h,48 h and 72 h were (41.909±4.333) ％,(54.470±3.756) ％ and (65.835±1.024) ％,respectively,and the apoatosis rate was (26.800±2.081) ％.Compared with 2 controls,the experimental group differences had statistically significance (P ＜ 0.05).Conclusion siRNA targeting HOXA9 can effectively silence HOXA9 gene expression in U937 cell,suppress cell proliferation and induce cell apoptosis obviously,which providing experimental basis for clinical lenkemia therapy by targeting HOXA9 gene.

ObjectiveTo estimate the applications of ADC value and rADC value in the diagnosis of nodular lesions of breasts.Methods Fifty-two cases with 66 nodular lesions of breasts confirmed by histopathology underwent diffusion-weighted magnetic resonance imaging.Three b values (0,800 and 1000 s/mm2) were applied.The mean ADC values of the breast nodules,the ADC values of ipsilateral breast( rADC1 )and ADC values of contralateral breast (rADC2 )were respectively measured.The independent-samples t-test and chi-square test were used for statistical analyses.ResultsOf the 52 patients,there were 18 patients with infiltrating ductal carcinoma and 34 patients with fibroadenoma.50 patients with 64 lesions were examined by DWI.( 1 ) at b = 800 s/mm2,the mean ADC values of malignant nodules [ ( 1.01 ±0.09) × 10-3 mm2/s],rADC800-1 (0.52 ±0.07)and rADC800-2 (0.51 ±0.06) were lower than that of the benign nodules [ ADC value = ( 1.54 ± 0.28 ) × 10 -3 mm2/s,t = 8.217,P ＜ 0.01 ; rADC800-1 =0.77 ±0.15,t =9.339,P＜0.01 ; rADC800-2 =0.76 ±0.14,t = 10.394,P ＜0.01 ].The one-side upper limits of 95％ medical reference value of mean values of infiltrating ductal carcinoma were adopted as the threshold point to distinguish the malignant from the benign.The threshold value of breast malignant nodule ADC,the rADC800-1 and rADC800-2 were respectively 1.05 × 10-3 mm2/s,0.55 and 0.53.The sensitivities of the three methods were 75.0％,65.0％ and 60.0％ ; the specificities were 100.0％,95.7％ and 97.8％ ;the positive predictive values were respectively 100.0％,86.7％ and 92.3％ ; the negative predictive values were 90.2％,86.3％ and 84.9％; the diagnosis accordance rates were respectively 92.4％,86.4％ and 86.4％.( 2 ) at b = 1000 s/mm2,the mean ADC values of malignant nodules [ ( 0.93 ± 0.08 ) ×10-3 mm2/s],rADC1000-1 (0.53 ±0.09) and rADC1000-2 (0.52 ±0.07) were also lower than that of the benign nodules[ ADC value= (1.45 ±0.28) ×10-3 mm2/s,t=11.844,P＜0.01; rADC1000-1 =0.75 ±0.16,t=5.820,P ＜ 0.01 ; rADC1000-2 = 0.74 ± 0.15,t = 8.082,P ＜ 0.01 ].The threshold value points breast malignant nodule ADC,the rADC1000-1 and rADC1000-2 were respectively 0.97 × 10-3 mm2/s,0.58,0.55.The sensitivities were all 70.0％ ; the specificities were respectively 100.0％,95.7％ and 93.5％ ;the positive predictive values were 100.0％,87.5％ and 82.4％ ; the negative predictive values were 88.5％,88.0％ and 87.8％ ; the diagnosis accordance rates were 90.9％,87.9％ and 86.5％ respectively.There were no significant differences in specificities and the diagnosis accordance rates ( x2 = 1.232,2.263 ; P =0.942,0.812 ).Conclusions ADC value and rADC value are both important parameters of MRI in differentiating benign and malignant breast diseases.The study indicated that ADC value ( at b =800 s/mm2) was the most valuable parameter.

Objective：To explore the characteristics of behavioral problems and self-concept level in children with attention deficit hyperactivity disorder(ADHD)accompanying anxiety disorder.Methods：With stratified random sampling method,a sampling survey was made in 9495 children(aged 5～17 years)from 6 regions of Hunan province together.Using the two stage epidemiologic research method,three groups,including control group(214 cases),pure ADHD(219 cases)and ADHD accompanying anxiety disorder(111 cases)were determined as the subjects.All the subjects completed the Children's Self-Concept Scale(CSCS),and the Achenbach Child Behavior Checklist(CBCL)was completed by their parents.Results：(1)Compared to the control group,the pure ADHD group scored higher in CBCL [e.g.,the total score:(30.35±24.32)vs.(16.97±14.86)],and scored lower in CSCS [e.g.,total score:(50.85±9.98)vs.(57.46±10.80),P<0.05].(2)Compared to the control group,children with ADHD accompanying anxiety disorder scored higher in CBCL [e.g.,total score:(41.66±28.07),P<0.01],and in CSCS except physical activities(Ps<0.05).(3)Compared to the pure ADHD group,ADHD accompanying anxiety disorder group scored higher in CBCL and lower in CSCS[behavior:(10.44±3.17)vs.(11.55±2.80);anxiety:(8.06±2.61)vs.(9.21±2.61),gregariousness:(7.42±2.22)vs.(8.16±1.89);total score:(47.25±9.93)vs.(50.85±9.98);Ps<0.05].Conclusion：ADHD children have more behavioral problems and lower level of self-concept than normal children,and children with ADHD accompanying anxiety disorder have more prominent presence of behavioral problems and lower self-concept rating than children with pure ADHD.

Objective To evaluate the value of enzyme-linked immunospot assay (TB ELISPOT) combined with serum latex agglutination test (LA) for diagnosis of pulmonary tuberculosis plus pulmonary cryptococcosis.Methods Serum and biopsy specimens of 76 patients, who were suspected of pulmonary tuberculosis and/or pulmonary cryptococcosis based on clinical and imaging features, were collected from March 2006 to September 2008 in Shanghai Public Health Clinical Center. TB ELISPOT assay, LA and histopathological examination were performed in all the patients. Results Histopathological and pathogenic examination confirmed pulmonary cryptococcosis in 15 cases and pulmonary tuberculosis in 22 cases, pulmonary tuberculosis plus pulmonary cryptococcosis in 8 cases. The sensitivity and specificity of TB ELISPOT were 91% and 94.4%. The sensitivity and specificity of LA were both 100%. TB ELISPOT assay and LA test were both positive in the 8 cases of pulmonary tuberculosis plus pulmonary cryptococcosis.Conclusions The value of enzyme-linked immunospot assay combined with serum latex agglutination test is high for diagnosis of pulmonary tuberculosis plus pulmonary cryptococcosis.