Objective To construct pcDNA3 1(+)GDNF recombinant eukaryotic expression plasmid and to investigate its expression in eukaryotic cells. Methods The coding sequence of GDNF was amplified from rat astrocytes by reverse transcription PCR (RT PCR) and cloned into pcDNA3 1(+) eukaryotic expression vector The recombinant pcDNA3 1(+)GDNF plasmid was then transfected into eukaryotic cells mediated by using Fu Gene 6 method Analysis by restricting enzyme digestion and DNA sequencing were carried out to demonstrate the sequence of the plasmid GDNF protein and its activity were then determined using pcDNA3 1(+)GDNF plasmid transfected eukaryotic cells Results RT PCR product is 640 bp specific segment Analysis by restricting enzyme digestion and DNA sequencing of pcDNA3 1(+)GDNF recombinant showed results from restricting enzyme were 640 bp and 300 bp segments respectively DNA sequencing revealed that GDNF cloning was successful The recombinant plasmid can express active GDNF protein in eukaryotic cells Conclusion The study on the role of both GDNF and gene therapy is significant in the treatment of Parkinson disease

Objective To investigate the relationship between the biological characteristics and the expres-sion of stromlysin (MMP-7) and microvessel density (MVD) in breast cancer tissue and the effect of MMP-7 on MVD of breast cancer. Methods 60 fresh samples were obtained from patients with breast cancer,and then the ex-pression of MMP-7 and vascular endothelial cell CD34 were studied by means of immunohistochemical assay (SP method). The relationship between MMP-7, MVD and age, tumor size, histological type, lymph node metastasis and expression of ER, PR, PCNA, p53 and CerbB2 was studied. The relationship between the biological characteristics and the expression of MMP-7 and MVD in breast cancer were evaluated. Results The positive expression rote of MMP-7 in the d>2 cm group,the group with lymph node metastasis,the group with positive expression of PCNA, p53 and C-erbB-2 (78.1%,74.2%,71.8%,67.6% and 72.2%)were all higher than that in the d≤2 cm group, the group without lymph node metastasis,the group without positive expression of PCNA,p53 and C-erbB-2(32.1%, 41.3%,38.1%,38.5% and 33.3%) (P<0.05). MVD was higher in the positive breast cancer tissue of the group of d>2 cm(34.61±6.97), the group with lymph node metastasis (34.37±7.50), PCNA ( 33.24±8.39), p53 (33.28±8.94), C-erbB-2 (33.55±8.57) than in the negative breast cancer tissue of the d≤2 cm group ( 28.60±9.82), the group without lymph node metastasis (27.48±8.66), PCNA (26.88±7.89), p53 (21.71± 7.59),C-erbB-2(27.42±27.69) (P<0.05). MVD(33.62±7.36)/ high power lens in the MMP-7 positive group was higher than that (27.86±9.45)/high power lens in the MMP-7 of negative group(P<0.05). The posi-tive expression of MMP-7 was correlated with MVD (r=0.380, P<0.05). Conclusion The high expression of MMP-7 and MVD is associated with the development and metastasis of breast cancer. MMP-7 could promote the mi-crovessel development in mammary cancer.

BACKGROUND:Colagen materials have good biocompatibility and biodegradability, but also had some problems such as low mechanical strength, poor resistance to degradation exposed in the process of clinical application. Numerous studies have reported that proper crosslinking could improve the disadvantage of colagen materials, regulate porous network structure, sweling and degradation of colagen materials. OBJECTIVE: To optimize carbodimide crosslinking process of colagen sponge and determine the best process conditions. METHODS:Colagen sponge was cross-linked by carbodimide for the preparation of loose and porous colagen sponge. Meanwhile, we optimized the conditions of cross-linking, in which the selected concentration of carbodimide was 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 mmol/L, linking time was 2, 4, 6, 8, 12, 16, 20, 24 hours, and linking temperature was 5, 10, 15, 20, 25, 30, 35℃. We evaluated the best process conditions of colagen sponge through the aperture, porosity, water absorption, and degradation rate. RESULTS AND CONCLUSION:The optimal conditions were carbodimide concentration 50 mmol/L, crosslinking temperature 20℃,crosslinking time 6 hours. At this point, the average pore diameter of colagen sponge was 105 μm, the porosity was 79.45%, water absorption was 287.14%, and the degradation rate was 15.04% (2 days). The crosslinking of colagen sponge significantly improved its water absorption and degradation resistance.

Objective To preliminarily analyze the credibility and validity of Chinese Medical Constitution Questionnaire(CMCQ).Methods CMCQ was used to investigate the distribution of Chinese medical constitution types of 450 women inpatients with postoperative nausea and vomiting (PONV) after gynecologic laparoscopic surgery from March to October of 2013.Credibility and validity were analyzed by Cronbach's α coefficient and factor analysis.Results Mixed constitution was identified in 184 (40.9％) patients.Cronbach's α coefficient of two dimensions (yang deficiency and qi stagnation) of CMCQ was proper,being 0.815 and 0.803 respectirely,while that of the dimension of harmony type was the lowest (0.514).The total variance explained by 9 common factors extracted from the results of factor analysis was only 48.5％.Conclusion CMCQ may need further revision based on the results of clinical application.

0.05). The interobserver variability in DSCT measurements was lower than in RT-3DE measurements in indexed left atrial volume and function. Conclusion DSCT can provide not only the information of coronary artery,but also highly reliable measurements of left atrial phasic volume and function at the same time.

Objective To apply the microbial transformation to transforming the total saponins in the fruit of Panax ginseng (SFPG) and preparing ginsenoside compound K (C-K). Methods The four microbial strains m14, m3, m8, and m9 were screened and isolated from the soil in the botanic garden planted for P. ginseng and they were used for the microbial transformation of SFPG to optimize the strains. Taking C-K contents as index the microbial transformation was detected and analyzed by TLC and HPLC. Results The strain m14 was found to transform the SFPG efficiently to C-K at first. The optimal culturing and transformation conditions of m14 were obtained: time, 6 d; temperature, 30 ℃; revolution of cradle, 160 r/min; initial pH value, 5.5; substratum concentration, 120 mg/mL. Under the optimal condition, the content of C-K was 41.65 times as much as before transformation by m14. Conclusion The m14 is the most effective strain among the four fungal strains. It is the new way available for the C-K industrialized production.

Objective To study the mechanism of enhancing effect of pretreatment with Geqin Tablets (GT). Methods The rat preconditioning models of myocardial ischemia were established by an improved pushing tube method. Cell apoptosis was determined with TUNEL. The expression of PKC was examined by immunohistochemical method. The serum SOD activity was tested by hydroxylamine hydrochloride method and MDA activity by TBA method. Results The amount of apoptotic cells increased in ischemia /reperfusion (IR) group significantly(P

Objective: To study the preparation technique and release characteristic of 5- fluorouracil-loaded chitosan microspheres for the intranasal administration. Methods:Using the liquid paraffin as the oil phase,and span-80 as the emuifier; 5- fluorouracil-loaded chitosan microspheres were achieved by emulsion-chemical crosslink technique. The orthogonal experimental design was applied to optimize the preparation procedure. Dynamic dialysis method was used to determine the releasing characteristic of microspheres in vitro and it influencing fators.Swelling behavior was expressed by swelling ratio.The degree of mucoadhesion was investigated by determining the mucociliary transport rate(MTR) of the microparticle across a frog palate. Results: Microspheres with a good shape and narrow size distribution were prepared. The average diameter was (43?4) ?m. The drug loading was 38.5%? 1.0%. The entrapment efficiency was 79.0%?1.8%.The drug release profile in vitro could be described by Higuichi eqution Q=0.1035t 1/2 +0.0284 (r=0.9965). Chitosan had good mucoadhesive property and caused a siginificant reduction in MTR(P

0.05), respectively. Conclusion: IgA1 could bind to HUVEC directly, and the binding capacities of DesIgA1 and Des/DeGalIgA1 to HUVEC were significantly higher than that of normal IgA1. Serum IgA1, especially deglycosylated IgA1, might play some role in vascular lesions of IgAN.

Objective To investigate the mutations of CLCNKB gene in a family with classic Bartter syndrome. Methods Genetic DNA was extracted from peripheral blood leucocytes of family members.The coding exons and intron exon junctions of CLCNKB gene were amplyfied by PCR and sequenced directly.Fifty unrelated healthy subjects were selected to exclude the possibility of polymorphism. Results A heterozygous(missense)mutation(482T>G,L161R)was detected in the exon 4 of patients.The hetemzygous mutation(L161R)was found in the mother,while no mutation was found in the father of this family.L161R had not been reported and was a novel mutation when referring to literatures and human genomic database home and abroad.Conclusion A new CLCNKB gene mutation(L161R)is identified for the first time.