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.

Nicotiana tabacum is an important and classical model plant which can respond to the change of environmental conditions by accumulating osmoprotectants, such as glycerol and proline which contribute to the re-establishment of homeostasis when exposed to various adverse environmental stresses, such as drought, salinity, high and low temperatures. The optimization of ultrasonic extraction (UE) conditions of glycerol-3-phosphate dehydrogenase (GPDH) of tobacco leaf have been built by orthogonal test. It showed that optimum of the powers, treatment times, slot times and leaf-to-solvent ratios of UE was 75w, 2h, 2s, and 1[DK]∶12 g/ml, respectively. Under these conditions, the activity of GPDH has been tested as 0.3937U/mg protein, which was higher than other extraction methods such as liquid nitrogen and grinding on ice bath. According to investigation, it is the first description of determination of content of GPDH with ultrasonic in tobacco. It could provide basis for the further research in the relation of content of glycerol and osmotic pressure in tobacco.

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.

Europe ; Hepatitis C ; diagnosis ; therapy ; Humans ; Practice Guidelines as Topic ; Societies, Medical

American Medical Association ; Humans ; Liver Failure, Acute ; diagnosis ; therapy ; Liver Transplantation ; Practice Guidelines as Topic ; United States

Objective To study the histone acetylation level of CD4+ T cells in peripheral blood of lupus nephritis ,to explore the role of histone acetylation in pathogenesis of lupus nephritis .Methods According to Feng X ,Bernstein ,Wagner S J and other scholars′s classification criteria for LN ,those who met the following conditions considered for the activity of LN :proteinuria >0 .5 g/d ,change or activity of urinary sediment (hematuria > 5 red blood cell /Hp ,or pyuria > 5 Hp white blood cells ,or 1 cell type/Hp) ,serum creatinine increased >1 .2 mg/L ,and the exclusion of infection ,kidney stones and other causes .Lupus nephritis pa‐tients were divided into inactive group (group I) 8 people ,active group (group A) 10 people .18 patients with LN and 8 normal con‐trols were collected peripheral blood 50 mL ,density gradient centrifugation method (Ficoll method) for separation of mononuclear cells in peripheral blood;CD4+ T cell was analyzed by immunomagnetic beads ,extracted histone acetylation level and detected H3/H4 protein by the acetylation of histone H3/H4 kits and the relationship of histone H3/H4 acetylation with diseases was analyzed . Results First ,compared with group N ,the histone H3 and H4 of CD4+ T cells in peripheral blood both in A and I of group LN pa‐tients showed low acetylation status (P<0 .01);Second ,the acetylation level of histone H4 in group A was lower than that in group I (P<0 .01) ,histione H3 acetyl the level of the group of two groups had no statistical significant (P>0 .05) .Third ,it was nega‐tively related to the acetylation level of histone H4 and 24 h urinary protein excretion(r= -0 .661 ,P<0 .05) .Conclusion Histone H3 and histone H4 of the CD4+ T cells showed low acetylation may be involved in the pathogenesis of lupus nephritis .The acetyla‐tion level of histone H4 in CD4+ T cells may be related to the activity of the LN .

Objective To observe liver pathological characteristics of the autoimmune hepatitis (AIH) C57BL/6 mice.Methods We established AIH C57BL/6 mice model with mixed adjuvants of allogenic liver antigens and complete freund's adjuvant by intraperitoneal injection.Then we determined and compared the body weight,alanine aminotransferase (ALT),aspartate aminotransferase (AST) between the normal control group (N group) and AIH model group (M group).And we compared the pathological characteristics of liver,spleen,heart,lung,kidney between two groups.Results We successfully established AIH C57BL/6 mice model.Compared with N group,we found that the average weight increase of mice in M group was decreased,ALT and AST of mice in M group were boosted.In M group,liver of mice presented typical interface of hepatitis,lymphocytic infiltration,even severe hepatitis,and showed spotty necrosis,multi-acinar necrosis.Some showed early sign of liver fibrosis by aprearing fibrous tissue hyperplasia.M group mouse's spleens were enlarged significantly.The spleen had darker color,not neat,and not smooth.Meanwhile,results were analyzed with statistics to confirm whether there was a significant difference between two groups (P＜0.05).About lungs,hearts and kidneys of mice in two groups,the pathological features were not found,and there was no statistic difference (P＞0.05).Conclusions The liver pathological characteristics of AIH C57BL/6 mice are similar to AIH patients.The results provide the pathological basis for the experimental research of autoimmune hepatitis.