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.

Cestode Infections ; parasitology ; Humans ; Infant ; Male

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 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.

Objective To determine the relationship between normal weight obesity (NOW) and cardiovascular risk factors.MethodsA total of 940 adults who received a health examination in out hospital were recruited in a cross-sectional study,and 407 with a body mass index (BMI) of18.5 to 25 kg/m2were enrolled for further analysis.Body fat percentage ( BF％ ) was measured by bioelectrical impedance analysis (BIA),and the subjects were assigned to the NOW group ( BF％ ≥25％ for male or BF％ ≥35％for female) or the control group ( BF％ ＜ 25％ for male or BF％ ＜ 35％ for female).Cardiovascular risk factors and their detection rates were compared between the two groups by using independent sample t test and x2 test.The correlationbetweenNOW and cardiovascular risk factors was assessedbylogistic regression.Results The prevalence of NOW in men and women were13.1％ and14.9％,respectively.The prevalence of NOW was increased with age ( x2 =6.90,P ＜0.05 ).Systolic blood pressure (SBP),diastolic blood pressure ( DBP ),total cholesterol ( TC ),triglycerides ( TG ),low-density lipoprotein cholesterol ( LDL-C) and serum uric acid (SUA) were significantly increased in the NOW group (t values were 2.97,2.44,2.54,5.09,2.71and 3.91,respectively; all P ＜ 0.05 ) ; whereas high-density lipoprotein cholesterol ( HDL-C) was significantly decreased in the NOW group (t =-3.90,P ＜ 0.05 ).The prevalence of hypertension,hyperglycemia,high triglyceride,low HDL-C,dyslipidmia and hyperuricemia was increased in the NOW group in comparison with the control group ( x2 values were 6.76,5.58,14.50,11.97,10.97 and 8.76,respectively;allP＜ 0.05 ).LogisticregressionshowedNOWincreasedtheriskof hypertension,hyperglycemia,dyslipidmia or hyperuricemia by 2.186,2.120,2.088 or 4.175 times.After adjustment for age and gender,the risk for hyperuricemia was decreased to 3.491,but remained statistically significant higher.Conclusions NOW may be correlated with cardiovascular risk factors,and those with NOW could be at higher risk for cardiovascular diseases.

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 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.