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.

Alzheimer’ s disease ( AD) , the most prevalent type of neurodegenerative disease, characterized by extracellular β-amyloid ( Aβ) plaque deposition, intracellular neurofibrillary tangles, and the loss of diverse neurons. Neuron loss might be the main cause that induces irreversible decline of cognitive function in AD patients. At present, AD therapy only relieves symptoms instead of fundamentally affecting the major pathologi-cal characteristics of the disease. Recently, rapid advances in neurogenesis and stem-cell biology have provided a new and pro-spective potential for AD treatment. Stem-cell types in the treat-ment of neurodegenerative diseases include neural stem cells ( NSCs ) , embryonic stem cells ( ESCs ) , mesenchymal stem cells ( MSCs ) , and induced pluripotent stem cells ( iPSCs ) . Stem cell therapy shows anti-AD function by replacing the dam-aged and lost neurons, rebuilding the cellular loop, inhibiting amyloidogenesis, promoting neurotrophic factor release, and reg-ulating immune reaction. This review highlights the recent pro-gress, mechanisms, and preclinical evidence of exogenous trans-planted stem cell, and addresses the current major challenges of stem cell transplantation in clinic. In addition, this review also summarizes the research status of compound promoting endoge-nous neurogenesis in brief.

Objective To investigate the influencing factors of chronic brucellosis.Methods Nested case control method was used to study newly diagnosed patients (n =600) with brucellosis in a cohort study in 2012.Data of general characterstics,clinical presentation,treatment and prognosis of those patients were collected.These patients were followed up for one year,and the chronic patients as the case group (n =248) and the healed patients as a control group (n =260).By means of Logistic multivariate analysis,factors turned brucellosis into chronic were screened.Result The chronic brucellosis-related factors were:gender,veterinary or epidemic prevention staff,muscle and joint pain,fatigue symptoms,and substandard treatment (x2 =5.163,16.445,14.977,17.154,8.813,all P ＜ 0.05).Conclusion Gender (female),veterinary or epidemic prevention staff,muscle and joint pain,fatigue symptoms,and substandard treatment are probably the chronic brucellosis-related factors

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 investigate the prevalence of anti-endothelial cell antibodies(AECA) and its possible role in the pathogenesis of propylthiouracil (PTU) induced ANCA positive vasculitis. Methods: Sera from 11 patients with PTU induced ANCA positive vasculitis and 10 patients with PTU induced ANCA but without clinical vasculitis were studied. Soluble proteins from in vitro cultured human umbilical vein endothelial cells were used as antigens and immunoblotting technique was performed to identify the specific target antigens. Results: In patients with PTU induced ANCA positive vasculitis group, 10 of the 11 patients in active phase were AECA positive and 7 of the 10 patients turned to negative in remission. AECA consisted of a group of heterogeneous antibodies. In patients with ANCA positive but without vasculitis, none was AECA positive. Conclusion: AECAs recognizing a variety of antigens could be found in sera from patients with PTU induced ANCA positive vasculitis and they had a much closer association with vasculitic disease activity compared with ANCA.

Objective: To investigate the relationship between the Smad4-independent pathway of TGF-?1 and drug-resistance of pancreatic cancer. Methods: The sensitivities of Smad4 homozygous-deleted pancreatic cancer cell line BxPC3 to different kinds of anti-cancer drugs (5-Fu, Gemzar, Oxaliplatin, Cisplatin, CPT-11 and Epirubicin) were observed by MTT assay before and after they were transfected with full-length cDNA of TGF-?1 or treated with TGF-?1 (5 and 10 ng/ml) solution. Western blot was used to detect p170 protein expression after stimulation with different concentrations of TGF-?1. Results: Cisplatin had the most powerful killing effect on BxPC3 cells, followed by Oxaliplatin, 5-Fu and CPT-11 with moderate effect and Gemzar and Epirubicin with the least effect. Cells transfected with full-length cDNA of TGF-?1 or treated with TGF-?1 solution became less sensitive to Cisplatin. Western blot revealed upregulation of p170 expression by TGF-?1. Conclusion: The Smad4-independent pathway of TGF-?1 can increase the drug resistance of pancreatic cancer cells through upregulating expression of p170.

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