Objective To construct lentiviral vector carrying rat′s calcitonin gene-related peptide(CGRP) gene for the following-up study on the function of CGRP .Methods CGRP gene segment was subcloned into shuttle plasmid ,become Puc57-CGRP .The pLenO-DCE-CGRP expression vector was be constructed by double digests .The pLenO-DCE-CGRP and 4 auxiliary packaging plas-mids were co-transfected into 293T cells .Cells were cultured for 48 hours .The supernant was collected and concentrated ,and then the viral titers were tested by multiple proportions dilution method and flow cytometer .The expression levels of CGRP were detec-ted in CGRP-modified 293T cells by Real-time PCR .Results The results of digestion and sequencing show that the pLenO-DCE-CGRP vector was constructed successfully .The titer of the lentiviral particles was 5 .1 × 108 TU /mL .Conclusion The high-titer lentvirus vector containing CGRP gene is constructed successfully ,which lay a foundation for transfecting mesenchymal stem cell (MSC) and studying the function of CGRP .

Objective To explore the relationship of acute myeloid leukemia kayotype with chemotherapeutic effect. Methods Conventional cytogenetic technique of G-band was used to analyze in 222 acute myeloid leukemia patients. Results Eighty-six cases (42. 1%) with cytogenetic abnormalities were found in 204 patients. Excepting for M_3 patients,the favorable,intermediate and adverse groups according for MRC criteria were treated with TA or CAG schema respectively. CR rate of M_3 was obviously higher than non-M_3. The favorable and intermediate group were obviously surpassinger than adverse group in TA schema. CR rate of the intermediate group was highter than that of other groups in CAG schema. CR rate of the adverse group in two schema were all low. CAG was surpassinger than TA schema in intermediate group. Conclusion Cytogenetics is an important chemotherapeutic and prognostic factor to acute myeloid leukemia. CAG schema could be the leader selection in the de novo presby or oligoac-crementional AML patients.

Professional features,which include taking students as its main body and improving the students' practical and operational ability as its aim are embodied in microbiology experimental teaching to improve the teaching quality and train high medical talents with different majors.Meanwhile,the design is intended to make medical graduates with different majors to quickly combine the microbiology knowledge with the practical situation in work thus to better serve the society.

Objective To explore the relationship of acute myeloid leukemia kayotype with chemotherapeutic effect. Methods Conventional cytogenetic technique of G-band was used to analyze in 222 acute myeloid leukemia patients.Results Eighty-six cases(42.1%) with cytogenetic abnormalities were found in 204 patients.Excepting for M3 patients,the favorable,intermediate and adverse groups according for MRC criteria were treated with TA or CAG schema respectively. CR rate of M3 was obviously higher than non-M3.The favorable and intermediate group were obviously surpassinger than adverse group in TA schema.CR rate of the intermediate group was highter than that of other groups in CAG schema. CR rate of the adverse group in two schema were all low. CAG was surpassinger than TA schema in intermediate group.Conclusion Cytogenetics is an important chemotherapeutic and prognostic factor to acute myeloid leukemia. CAG schema could be the leader selection in the de novo presby or oligoaccrementional AML patients.

OBJECTIVE：To explore the possible mechanism of astragalus in improving cardiac function.METHODS：To observe the effects of astragalus,added to basic anti-heart-failure therapy,on cardiac function and TNF in 45 cases of thronic heart failure.RESULTS：In comparison with basic therapy group,the improvement of cardiac function was more obvious and the level of serum TNF-α was lower in astragalus group.CONCLUSION：Astragalus probably has the action of decreasing TNF resulting in improvement of cardiac function.

Objective To explore relationship between resistin'adipenectin and insulin resistance(IR) in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS). Methods Based on polysomnography (PSG) and disease history ,68 patients were divided into three groups: mild ( L, n = 24 ), moderate ( M, n = 13 ), severe ( S, n = 31 ) OSAHS, and normal control ( n = 20 ). Fasting serum glucose, insulin, lipoids, resistin and adipanectin were measured. HOMA-IR was calculated, the correlation between HOMA-IR and resistin, adiponectin, lipoids, body mass index, waist-to-hip ratio, apnea-hypopnea index ( AHI), the lowest saturation of blood oxygen ( LSaO2) and the per-centage of total sleep time spent with an oxygen saturation less than 90% (T90) were analysed. Results The levels of serum resistins in each OSAHS group were (8.04 ±2. 14), ( 10.85±4.89 ), ( 13.34±3.52 )mg/L, and were sig-nificantly higher than those in control group ( 9.49 ± 2.40) mg/L ( P < 0.05 ), S were significantly higher than L (P<0.05) ;The levels of serum adiponectin of OSAHS groups were(6. 21 ± 1.74), (4. 19±1.80), (2.26± 1.17 ) mg/L, and were significantly lower than those in control group(9.49±2.40) mg/L (P < 0.05 ), and the inter-group differences were statistically significant (P <0. 05). HOMA-IR of M and S were(4.07±0.97), (5.61± 2.26) significantly higher than those of L and control group( 1.57 ±0. 58 ), (2.47 ±1.52 ) ( P < 0.05 ), and inter-group differences between M and S were statistical significant( P < 0.05 ). Spearman correlation analysis showed that HOMA-IR was significantly positive correlated with resistin, total cholesterol, triglyceride, body mass index,waist-hip rafio,AHI,T90( r =0. 794,0.438,0. 430,0. 351,0. 456,0.775,0.624 ,P <0.01 ) ,negative with adiponectin, LSaO2 (r=-0.563, -0.623 ,P <0.01 ). Partial correlation analysis showed that after the effect of adiponeetin and resis-tin were controlled,HOMA-IR was significant positive correlation with T90 and AHI(r =0.231,0.358 ,P <0.05 ). Multiple stepwise regression analysis showed that, to HOMA-IR, resistin and AHI were the most significant impact factors (R2 =0.613,F=69.810,P<0.01). Conclusions HOMA-IR is significantly positively correlated with the severity of OSAHS,independent of the blood lipids, body mass index, waist-hip ratio, resistin, adiponectin and other factors;the effects of blood lipids,body mass index,waist-hip ratio and other factors on HOMA-IR probably depend on resistin and adiponectin;AHI and plasma resistin level may determine the level of insulin resistance in patients with OSAHS.

Objective To investigate the calculation method and its inlfuencing factors of Z scores in the aortic root diameters measured by echocardiography in children. Methods A total of 105 children with median age 19 months, who came to Shenzhen Children′s Hospital from March 2012 to October 2012 were included. The diameters of aortic ring (ARD) and aortic sinus (ASD) were measured by two dimension echocardiography, Z scores of ARD and ASD were calculated using two different normal reference values regression equation and mean square error derived from Shenzhen children′s hospital (C method) and Pettersen et al (P method). The regression equation from C method and body surface area (BSA) formula from P method were adopted to calculate Z scores of ARD and ASD (ZH method). The Z results of ARD and ASD calculated by those three methods were compared and were analyzed for their normality probability distributions. Results Z scores of ARD and ASD derived from C method were all showed as normal distribution (P=0.067 and 0.650). Z scores of ARD and ASD derived from P method were all showed as normal distribution (P=0.208 and 0.970). Z score of ARD derived from ZH method was showed as non-normal distribution (P=0.027), but Z score of ASD was normal distribution (P=0.430). There were no significant differences in ARD-Z calculated by C method (0.41±0.89), P method (0.23±0.85) and ZH method (0.36±0.94) (F=1.117, P=0.309). There were signiifcant differences in the Z scores of ASD calculated by C method (0.38±0.89), P method (0.58±0.71) and ZH method (0.36±0.84) (F=5.443, P=0.005). Z scores of ARD (r=0.917, P=0.000) and ASD (r=0.900, P=0.000) calculated by C method correlated well with that by P method. Conclusions Calculation method of BSA and normal reference values regression equation were the main influencing factors of Z score value in quantifying children aortic root diameters by echocardiography. For the clinical applications. The normal reference value should be used which is suitable for the Chinese children.

ObjectiveTo investigate whether neuronal intermediate filament protein(NF-66) could be used as a molecular marker for the determination of malignancy of insulinoma.MethodsThe expression of NF-66 protein was detected in insulinoma and pana - cancerous tissues and 3 insulinoma cell lines by immunohistochemistry staining. The relationship between the expression of NF-66 protein and the clinicopathological characteristics,survival was analyzed by univariate and multivariate statistical analysis.ResultsExpression of NF-66 was found in 102(77％ ) out of 132 insulinomas and none of 98 paired control (P =4.86 × 10-31 ).NF-66 was highly expressed in 3 insulinoma cell lines.The expression of NF-66 was found in 96 (81％) out of 118 benign tumors (P =0.003),while out of 14 malignant insulinomas,6(43％ ) were found to express NF-66 ( P =0.003 ).The expression of NF-66 was significantly associated with tumor size (3 cm as the cut-off point),distant metastasis (38％ vs 81％,P =0.013) and distant plus lymph node metastasis (46％ vs 81％,P =0.009),respectively.The expression of NF-66 was not correlated with age,gender,recurrence and overall survival.ConclusionsDown-regulation of NF-66 was significantly associated with tumor malignancy,suggesting that NF-66 could be a potentially novel molecular bionarker to distinguish malignancy from benign insulinoma.

Objective:To construct a eukaryotic expression vector of leptin gene,and to transfect it into human placental mesenchy-mal stem cells(HPMSCs)and appraise its expression.Methods:Primers were designed and the leptin gene was obtained by RT-PCR from human adipose tissue.The aimed segments were inserted into the eukaryotic expression vector pIRES2-EGFP,plasmid pIRES2-EGFP-LEP was constructed and identified by restricted enzymatic resection,and then transfected it into HPMSCs by liposome.The ex-pression of leptin in the transfected cells was detected by RT-PCR and Western blotting,the multi-differentiation potential of the cells was indentified.Results:The length of specific fragment was 500 bp,the recombinant plasmid pIRES2-EGFP-LEP presented 5.3 kb and 500 bp bands by restriction enzyme digestion.Leptin gene was expressed in transfected HPMSCs and the transfected HPMSCs maintained multi-directional differentiation potential.Conclusion:The eukaryotic expression vector of leptin can be transfected and ex-pressed in HPMSCs.

Objective To observe the effect of adenovirus- receptor activity modifying protein-1 (RAMP1) on nuclear factor(NF κB) and myocardial fibrosis in heart of rabbit with myocardial infarction. Methods Myocardial infarction (MI) models were developed in 54 rabbits and they were randomly divided into RAMP1 group,EGFP group and control group according to whether pAd2-EGFP-RAMP1,pAd2-EGFP or PBS was injected into infarction region and its border.At 7 d,14 d and 28 d after injection,Left ventricular function indices such as LVEF,LVSd,LVDd and LVFS were estimated by echocardiogram,the expression of NF-κB in myocardium was detected by Western blot,the plasma level of TNF-α was measured by ELISA,and fibrosis and collagen content was measured by Masson stain. Results At 7 d after adenovirus injection,the expression of RAMP1was significantly increased in RAMP1 group (67.33 ± 3.97)％ as compared to EGFP group(20.59 ±3.26) ％ and PBS group ( 23.80 ± 5.08) ％ ( P ＜ 0.05 ).The expression of NF κB was decreased in RAMP1 group ( 26.54 ± 5.13 ) ％ versus EGFP group (62.60 ± 6.18) ％ and PBS group (62.95 ±5.17)％ (P＜0.05).The plasma level of TNF-α was lower in RAMP1 group than in EGFP group and in PBS group at different time[7 d:( 136.74 ± 5.42) μg/L vs.( 196.97 ± 14.17) μg/L,(203.67 ±13.90)μg/L; 14 d:( 154.51 ± 13.61 )]μg/L vs.( 112.22±6.74 )μg/L,(160.46± 14.27)μg/L ;28 d;(51.10± 5.62)μg/L vs.( 95.55 ± 9.94 )μg/L,( 98.96 ± 12.68) μg/L,all P＜ 0.05].The collagen content was reduced in RAMP1 group as compared with EGFP group and PBS group at 14 d and 28 d [14 d:(7.10±0.98)％ vs.(19.52±2.32)％,(17.91±0.96)％ ;28 d:(17.04±2.44)vs,(34.10±5.59) ％,(33.98±4.33)％,all P＜0.01].At 28 d after infarction,the infarct size was decreased in RAMP1 group (26.54 ± 5.13) ％ compared with EGFP group (32.20 ± 3.73) ％ and control group (35.58±2.65) ％ (P＜0.01),and better heart function appeared in RAMP1 group. Conclusions The high-expression of RAMP1 could decrease the collagen deposition and fibrosis in the border of infarction and improve heart function through lower expression of NF-κB and decreasing the plasma level of TNF-α.