Objective To explore alterations of cognitive function and P300 in depressed patients with Parkinson's disease(PD).Methods Zung's self-rating depression scale(SDS) and Hamilton depression rating scale(HAMD) were used to divide the 67 patients with PD into two groups, PD with depression and PD without depression. The cognitive function and P300 were measured, further comparing and the correlation analysis were made.Results The score of Mini-mental state examination (MMSE) in the PD patients was in normal extent,but significantly lower than that in the normal controls(P

Aging ; psychology ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Learning ; drug effects ; Male ; Memory ; drug effects ; Rats ; Rats, Wistar

Objective:To obtain a large amount of erythromycin B and to investigate the activity site in eryK. Methods:The key sequence of the BC loop region in eryK gene was knocked out and the eryK gene with 101 bp deleted was amplified by overlapping PCR,and cloned into vector pWHM3 to construct recombinant plasmid. The Saccharopolyspora erythraea mutant AK17 was constructed through chromosomal homologous recombination technique.Results and Conclusions:The S.erythraea mutant AK17 was constructed. The results of TCL and MS analysis showed that the major fermentation product of AK17 is erythromycin B.

Background Diabetic retinopathy (DR) is a common ocular complication of diabetes,and its pathogenesis is associated with a variety of factors.c-Jun N terminal kinase (JNK),one of the genes involving in apoptosis,plays an important role in the pathology of diabetes,and relative research is catching increasing interests in recent years.Objective This study was to quantify the expression of JNK3 in retinas of DR murine.Methods Forty-eight SPF male C57BL/6 mice were randomly divided into the diabetes group and the normal control group.Diabetic mouse models were establishend by intraperitoneal injection of 1％ streptozocin (STZ) dissolved by sodium citrate buffer,and equvilant volume of sodium citrate buffer was used in the same way in the mice of the control mice.The left eyeballs were obtained 2,4,8 weeks after modeling and the retinas were collected.Real-time quantitaive PCR was perfored to detect the expression of JNK3 mRNA in retinas.The use and care of the experimental mice complied with the Administration of Experimental Animals in Kunming Medical College.Results Blood glucose levels were significantly higher in 2,4,8 weeks after modeling in the diabetic group compared with the normal control group (t=-5.675,-5.498,-5.347,all at P＜0.01).The relative expression levels of JNK3 mRNA (A value) in the retinas were significantly different between the groups at various time points (Fgroup =102.345,P＜0.05 ; Ftime =131.679,P＜ 0.05).The relative expression levels of JNK3 mRNA in the retinas were 3.21 ±0.14 and 5.43 ±O.37 in 4 and 8 weeks after modeling in the diabetic group,which were significantly elevated in comparison with the normal control group (2.54±0.42 versus 2.26±0.67) (t =4.073,23.399,both at P＜0.05).Compared with the second week and fourth week,the relative expression levels of JNK3 mRNA in the retinas in the eighth week were significantly raised in the diabetic group (t =10.756,16.857,both at P ＜ 0.05).Conclusions JNK3 expression in the retina upregulates in diabtic mice in a time-dependent manner.JNK3 is paopably involved in the pathogenesis and development of DR.

Cell viability of Pichia pastoris was detected by flow cytometry (FCM) with two reagents fluorescein diacetate (FDA) and propidium iodide (PI). Compared with FDA/PI double-stained dot plots and PI single-stained dot plots,the latter could divide dead and living cells into two separate zones,and get the correct proportion. Then PI single-stained method was used to detect the change of cell viability in Pichia patoris fermentation. At glycerol batch and fed-batch phase,little dead cells were detected. At methanol fed-batch phase,cell viability decreased when cell weight increased,and was only 73.8% at 88 h.

Proteolytic degradation has been a severe problem when Pichia pastoris is employed to express recombinant proteins.One alternative method to circumvent this problem is to construct protease gene disruptant.However,the main study of gene disruption is focused on nonrecombinant Pichia pastoris rather than recombinant strain.In our study,we established two different methods to directly disrupt PRC1 and KEX1 gene in recombinant Pichia pastoris.On the basis of this,we further discussed and compared the application and advantages of both methods.

The kinase domain of receptor tyrosine kinase(RTK) ErbB2 was expressed fused with GFP in Pichia pastoris. Recombinant expression vector pPIC3.5K was constructed in Escherichia coli TOP10. The right P. pastoris transformants were screened on his-deficient plates and YPD-G418 plates by turns after electroporation of recombinant vector, and then induced by methanol in baffled shake bottles. The strain with highest protein yield was scaled up in a 5 L fermentor. Recombinant protein was analyzed with tyrosine kinase assay after Ni2+ affinity chromatograph. Results showed that the 100 kD recombinant protein with tyrosine kinase activity was successfully expressed in P. pastoris.

The bioreactor production of recombinant Lateolabrax japonicus growth hormone (rljGH) expressed intracellularly by Pichia pastoris was investigated. A strategy of feeding methanol at the exponential rate was established and the effect of specific growth rate on the rljGH production was examined. The results indicated that the average specific production rate increased and the rljGH production duration decreased as the specific growth rate increased. The maximum specific rljGH production (0.58 mg/g WCW) was achieved at a specific growth rate of 0.029/h. The effect of supplementing ammonium sulfate, peptone and yeast ex- tract on the rljGH production was further investigated. The results indicated that the effects of ammonium sulfate and peptone were not significant. Supplementing yeast extract of 2.5 g/L was advantageous for the rljGH production. The duration of the rljGH production was increased to 23 h from 17 h and the fermenta- tion stability of run-to-run could be improved.

A fusion expression vector pPIC3.5K-PDGFR? was constructed to express recombinant receptor tyrosine kinase PDGFR? and the right Pichia pastoris transformants were screened on his-deficient plates and YPD-G418 plates by turns after electroporation of strain GS115, a high yield strain named M3 was screened. The strain M3 was cultured in a 5 L fermentor and His-GFP-PDGFR? fusion protein was purified by Ni2+ chelating affinity chromatography. One distinct peak was obtained after elution with 250 mmol/L imidazole. Fusion protein was proved to be 90.08 kD by western blotting, and have tyrosine kinase activity by ELISA. Results showed that the receptor tyrosine kinase PDGFR? was successfully expressed in P. pastoris and could be used as a target for small molecule selective inhibitors screening.

Objective To explore the effect of uric acid (UA) on the rat glomerular mesangial cells (GMCs) and its possible mechanism in vitro. Methods Cultured rat GMCs were treated with various concentrations of UA (50 μmol/L, 100 μmol/L, 300 μmol/L) in the presence or absence of U0126, apocynin, rotenone. The incorporation of 3H-thymidine (3H-TdR) and cell counting were used to measure GMCs proliferation. GMCs cell-cycle was analyzed by flow cytometry. CyclinD1 and cyclin A2 expression were determined by real-time PCR and Western blotting analysis. ERK1/2 phosphorylation was detected by Western blotting. Reactive oxygen species (ROS) was measured by 2, 7-dichlorofluorescein diacetate (DCFDA) fluorescence. Results (1) Uric acid increased GMCs proliferation in dose-dependent manner compared with the control groups, as assessed by 3H-TdR incorporation and cell counting. GMCs proliferation induced by 300 μmol/L uric acid was increased by more than 1.5-fold assessed by both of the methods. (2) Uric acid decreased cell number in G1/G0 phase and increased cell number in S phase in dosedependent manner, as assessed by flow cytometry. (3) Uric acid iuduced cyclin D1 and cyclin A2 expression in dose-dependent manner. (4)Uric acid increased pospho-ERK1/2 in dose-dependent manner and ERK1/2 specific inhibitor U0126 could suppress uric acid-induced cell proliferation.The inhibition percentage of U0126 was 22% and 31% assassed by cell counting and 3H-TdR incorporation, respectively. (5) Uric acid increased ROS production in dose-dependent manner.NADPH oxidase inhibitor apocynin could also significantly inhibit uric acid-induced ROS production, ERK phosphorylation and cell proliferation. In contrast, rotenone had no effect on them.Conclusion Uric acid can stimulate rat GMCs proliferation, partially by the activation of ERK pathway via NADPH oxidase-derived ROS generation.