The fused gene (PTH-HSA) of parathyroid hormone (PTH) gene and Human Serum Albumin(HSA) gene was amplified without linker by Overlapping PCR technology. The spliced gene was clone into Pichia pastoris secretory vector pPIC9K. With the help of promoter AOX1 and mat ? signal peptide, the PTH-HSA gene was designed to secretory expression.Linearized by restriction enzyme SalI, The recombinant plasmid pPIC9K/PTH-HSA was transformed into Pichia pastoris KM71 by electroporation. The recombinant strains which were identified by G418 and PCR analysis were induced by methanol to express protein PTH-HSA. The target protein was expressed in fermentation supernatant. Western blot analysis of the fusion protein showed that the expressed fusion protein PTH-HSA had the antigenicity of HSA.adenylate cyclase assay proved that the fused protein exhibited the bioactivity to stimulate cAMP synthesis The specific activity of broth was about 318IU/ml.

Objective To analyze the clinical and pathological characteristics of children with lupus nephritis(LN).Methods The data of 116 inpatients from Mar.2000 to Nov.2008 with LN were retrospectively analyzed.The clinical,immunochemical and pathological data were recorded.Renal tissue was observed by light microscopy and electron microscopy after HE,PAS,Masson and PASM staining according to WHO standards.Follow-up results showed complete remission,partial remission,disease activity,renal insufficiency and death.Results Of the 116 cases of LN,there were 27 male and 89 female with a ratio of male to female 1.03.3,and the mean age was(12.0?2.2) years.The incidence of nephrotic syndrome was 63.8 %(74 cases),and acute nephritis was 21.5%(25 cases).Class Ⅳ LN was the most frequent type(14 cases,50%) and classⅢ was next(25 cases,21.5%).In view of the outcome,rapidly progressive glomerulonephritis and class Ⅳ LN were the worst.LN was initially controlled in 96.5% of the patients.Relapses of LN were most caused by the intermittent treatment.Totally 32 cases showed different renal injury,and 6 cases progressed to death.Conclusions Renal biopsy is important to diagnosis,treatment and prognosis evaluation of LN.Long and regular treatment is important for children with LN.

Background Congenital heart disease (CHD) is a diverse group of diseases determined by genetic and environmental factors.Considerable research has been done on genes associated with the development of the heart.Recently,focus is on the role of transcription factor NFATc1 in the development of proper valve and septa.As part of a larger study,high density single nucleotide polymorphism (SNP) scanning was used to explore the relationship between NFATc1 gene polymorphism and susceptibility to ventricular septal defect (VSD) in the Chinese Han population.Methods One hundred and ninety-two pediatric patients with congenital VSD and 192 matching healthy control subjects were studied.The haplotype reconstructions were calculated by PHASE2.0 software.Haploview software was used to perform linkage disequilibrium assessment and define haplotype blocks.The algorithm used for defining the blocks was the confidence interval method.Results The NFATc1 gene region can be divided into 11 haplotype blocks.Strong linkage disequilibrium existed within blocks 6,8,9,and 11.Three SNPs (rs7240256,rs11665469,and rs754505) within the NFATc1 gene had significant correlation with VSD by single marker association analysis.In addition,two haplotypes correlated with VSD.Conclusions NFATc1 is associated with the occurrence of VSD and it may be a predisposing gene to CHD in Han Chinese.This finding has set a direction for further genetic and functional studies.

Objective To establish VX2 hepatic carcinoma model in rabbits by implanting the tumor fragment into the liver through percutaneous puncture under ultrasound guidance and to observe its growing and metastatic characteristics, to determine the optimal time for interventional experiment study with the model. Methods Inoculation of VX2 carcinoma fragment was performed in 28 New Zealand white rabbits. PET/CT and ultrasonography (US) examinations were carried out in the second, third and forth week after the inoculation, and each time two tumor-bearing rabbits were sacrificed for pathologic study. Results The successful rate of model establishment was 89.28% (25/28). On PET or CT scans, single lesion in the liver was demonstrated in 25 rabbits. Two, three and four weeks after the inoculation, the maximum diameter of the tumor was (4.82±0.80) mm, (16.05±2.89) mm and (30.08±5.38) mm respectively, while the metastasis rates was 0% (0/25), 13.04% (3/23), 76.19% (16/21) respectively. No significant necrosis was found in the second week after inoculation, only tiny coagulation necrosis was revealed in the third week, and massive necrosis was seen in the forth week. Conclusion Percutaneous inoculation of the tumor fragment into the liver under ultrasonographic guidance is a simple method to establish VX2 hepatic carcinoma in rabbits with a high successful rate. The third week after inoculation is the suitable time for making interventional experiment study.

A bacterium strain BFHM2002 is isolated from Lake Donghu, Wuhan. BFHM2002 has advantages that it can produce melanin with a high rate and high yield in the absence of tyrosine. Induced by tyrosine, melanin yield can be dramatically increased. BFHM2002 may be identified as a new strain in Bacillus firmus, for melanin-production.

The hom gene encoding for homoserine dehydrogenase was amplified from the genomic DNA of Corynebacterium glutamicum ATCC 13032.After the kanamycin-resistant gene(Km)cassette from plasmid pET28a was inserted into the center of hom,the hom::Km cassette was then electroporated into the competent cell of C.glutamicum ATCC 13032.And kanamycin-resistant clones were obtained.PCR was performed to confirm whether the Km gene was integrated into the hom gene of these clones and the recombinant strains of hom-disrupted were screened out.Fermentation results showed that the lysine yield of the hom-disrupted strain C.g-hom::Km-8 reached 4.7 g/L,which was 6.7 times that of C.glutamicum ATCC 13032.

In order to isolate genes related with the osmoadaptation and glycerol metabolism of Candida glycerinogenes, a transformation system based on the dominant selectable marker Zeocin and restriction enzyme-mediated integration (REMI) was established. Effects of seven restriction enzymes on transformation efficiency of C.glycerinogenes were tested. Transformation conditions were optimized in the presence of Hind III. Under the optimal conditions of OD_ 600 ≈1.3, voltage of 1.5 kV, 2.0?10~9 competent cells/mL, 100 units of Hind III added, the transformation efficiency was up to 129 trnaformants/?g DNA. 58% of transformants were stable on nonselective medium. These results suggest that REMI technique would be beneficial to the genetic transformation of C.glycerinogenes.

A method to determine dihydroxyacetone (DHA) in fermentation broth was developed by high performance liquid chromatography (HPLC). DHA was separated on a Alltima C18(5?m,250?4.6mm). The mobile phase was 0.5% methanol solution (pH adjusted to 3.0 with H3PO4), the flow-rate was 1.0 ml/min and the detective wavelength was 200 nm. The detection limits of DHA was 0.1 g/L～10.0 g/L. 6.2 g/L DHA in the fermentation broth was detected by HPLC, which was in agreement with the result by spectrophotometric method.The method was applicable for DHA determination in the fermentation process.

More than 20 strains capable of producing dihydroxyacetone from glycerol were isolated from 4 different natural environment samples by using two detection methods. The strain 6-8 which could grow on medium containing glycerol as sole carbon source had a higher converting capability. Under a better culture, the highest DHA production of the strain 6?8 reached 6.4 g/L. In addition to general morphological and bio-chemical characteristics, the strain 6?8 was identified by 16S rDNA sequence and systematic analysis. The results showed that 16S rDNA sequence of the strain 6-8 had similarity of 99.7% with Acinetobacter sp. suggesting that the strain 6-8 is one of subspecies of Acinetobacter sp.

The method for analysis and determination the cleavage of soybean sterol, in which the soybean sterol was degraded and the products androst-1,4-diene-,17-dione (ADD) and androst-4-ene-3,17-dion (AD) were developed by Liquid Chromatography-mass Spectrometry. The HPLC conditions adopted were: a All- tima ODS-2 column (250 mm?4.6 mm, 5 ?m), a mobile phase consisted of menthanol-water (70:30), a flow rate of 1.0 mL/min, a room column temperature. and the detective wavelength was 244 nm.The ZMD Micromass electrospray ionization (ESI)-mass spectrometer was employed. In such conditions the corre- sponding HPLC chromatogram and MS spectrum were obtained. The method has a linear ranger of 0.01 mg/mL ~ 0.09 mg/mL, R2 =0.9999, the recoveries of ADD and AD were 102.6% and 105.90%, the RSD of ADD and AD were 3.02%, 3.5% and 3.08%, 3.24%. This method showed high sensitivity, accuracyand easy to perform. It is suitable to analysis the process cleavage of soybean sterol as well as quality control of product.