AIM: To express the synthesized human insul in like growth factor I (hIGF-1) gene in E.coli with high expression level a nd explore the way to increase the efficiency of factor Xa cleavage. METHODS: The gene of hIGF-1 was designed and synthesized accordi n g to the preference of E.coli. A fusion protein with a recognized site of fa ctor Xa between CBD (cellulose binding domain) and hIGF-1 was expressed and puri fied by cellulose affinity chromatography. MTT method was used to assay the bioa ctivity of CBD-IGF fusion protein. hIGF-1 was released by factor Xa. In order to improve the sensitivity of fusion protein to factor Xa, the short flexible pept ide (Gly-Thr-Gly- Gly-Gly-Ser-Gly) was added before the recognized site of fac tor Xa. RESULTS: SDS-PAGE results indicated that the CBD-IGF fusion prot ein was expressed and purified . Biological assay results indicated CBD-IGF fusi on protein could promote the growth of NIH3T3 cell. The short flexible peptide (Gly-Thr-Gly-Gly-Gly-Ser-Gly), which was added before the recognized site of f actor Xa, improved the sensitivity of fusion protein to factor Xa. CONCLUSION: CBD-IGF fusion protein with bioactivite are expresse d and purified. The amio acid sequences changes between the site recognize of fa ctor Xa can help to improve the cleavage efficiency of Factor Xa.

Effects of squalene, algae (Enteromorpha prolifera), laver (Prophyra hai-tanensis) and cauliflower on serum total cholesterol (T-C), high density li-poprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and very low density lipoprotein cholesterol (VLDL-C) were separatly observed in Wistar male albino rats fed on a diet containing 0.5% cholesterol,5% lard and 0.5% bile salt. Squalene was supplemented at 5% of the diet, and algae, laver, cauliflower at 10% of the diet.After the rats had been fed on the diet described above for 30 days, serum T-C, HDL-C, LDL-C and VLDL-C levels were determined. The results showed that the rats fed on a diet containing 5% squalene or 10% algae were significantly lower in serum T-C, HDL-C and VLDL-C but higher in serum HDL-C than those of the controls, and the rats fed on a diet containing 10% laver or 10% cauliflower were significantly lower in serum T-C than that of the controls.

Z-Ligustilide, a major phthalide isolated from a widely used traditional Chinese medicine Ligusticum chuanxiong, possesses various pharmacological activities including neuroprotective, anti-inflammatory, antiproliferative and vasorelaxing effects. However, it is unstable and inclined to degrade in natural conditions, which limits its study and application greatly. In this study, degradation behavior of Z-ligustilide and its degradation products stored at room temperature under direct sunlight were investigated and structure elucidated by HPLC-UV, UPLC-QTOF-MS and NMR. Z-ligustilide degradation and total five degradation products were generated and detected. Two degradation products were unequivocally identified as senkyunolide I and senkyunolide H by comparison with reference compounds. Another two degradation products were further isolated by semi-preparative HPLC and structure elucidated as (E)-6, 7-trans-dihydroxyligustilide and (Z)-6, 7-epoxyligustilide by 1H and 13C NMR, respectively. The degradation pathways of Z-ligustilide were finally proposed. Oxidation, hydrolysis and isomerization are the major degradation reactions.

imaging features of myositis ossificans have some characteristics. Misdiaguosis could be avoided when the disease was evaluated with the course.

Objective To reflect the spatial distribution of brucellosis among 7 counties in Qinghai Province from 2011 to 2013 with Geographic Information System (GIS) map.Method Epidemiological situations were overlay analyzed in infectious rate and prevalence rate with QHEndemic-GIS and SPSS 17.0 by statistics and mapping.Results Brucellosis was detected in 7 counties through the spatial distribution map.The infectious rate and prevalence rate of brucellosis were showing a phenomenon of high and low alternation in 3 years;meanwhile the prevalence in Gonghe County showed an obviously upward trend with the rates of 0.50％ (1/201),1.50％ (3/200),and 3.73％ (9/241),respectively.Conclusions GIS could reflect the state of brucellosis infection directly and correctly.Prevalence of brucellosis in the 7 counties has showed a sporadic phenomenon,and the prevalence of brucellosis has covered a wide region and many populations are at risk of this disease.The prevalence has showed an up-ward trend in these regions,which provides a scientific basis for prevention and control of brucellosis in these key regions.

AIM: To study the effect of Liuwei Dihuang Wan (LWDHW) on the expression of immune-associated genes in the aged rats using cDNA microarray. METHODS: Forty 20-months-old rats were divided into two groups equally. One group was treated with LWDHW for 5 weeks,another was untreated as control. Some biochemical assays were used to confirm the physiological differences between two groups. The mRNA from the spleens of twenty 4-months-old young rats were extracted and divided equally into two parts. Each part was conjoined with mRNA from drug treated aged rats and that from untreated control aged rats,respectively,and two experimental combinations for reverse transcription and cDNA microarray were formed. By comparing two groups of data from two combinations,the effects of LWDHW on the expression of the associated genes were evaluated. RESULTS: 13 down-regulated genes and one up-regulated gene were identified in the untreated control old rats,whose expression did not change obviously in treated old rats compared with young rats. The expressions of another two sequence were up-regulated distinctly in treated old rats compared with young rats. CONCLUSIONS: 16 immune-associated genes expressions were affected markedly by LWDHW. It will be helpful for understanding the molecular mechanism of LWDHW in ageing.

We report herein the effects of varying dietary zinc contents on burns healing in 15% body surface area (BSA) Ⅲ? burned Lop rabbits. 18 adult male rabbits were given a low zinc diet for two weeks, resulting in a decrease of serum zinc concentration which further decreased postburn without restoration. On the contrary, serum was restored rapidly to normal levels in the zinc supplemented groups. Similar changes were found in serum alkaline phosphatase (AKP) activity, and there was a close relationship between serum zinc and serum AKP activity. Burns healing in the high zinc group was markedly faster than that in the low zinc group. These results strongly suggested that zinc might be a factor benefiting burns healing. We concluded that serum AKP activity could be used as a diagnostic criterion of postburn hypozincemia as well as a valid index in the assessment of the effectiveness of supplemented zinc.

Objective To observe the impact of transplant nephrectomy on panel reactive antibodies and a secondary renal transplantation.Methods Panel reactive antibodies in 15 patients with a failed renal transplant admitted in our hospital between 2004 to 2007 were measured before transplantation,before and 1 month,6 months,12 months after transplant nephrectomy,and the pathological changes were observed after transplant nephrectomy.Results Panel reactive antibodies was increasing after renal transplantation,and reached the highest level one month after transplant nephrectomy,then grandually got down.New HLA allosensitization sites were discovered after transplant nephrectomy.Large amount of C4d was stained in failed transplant.Conclusion Serum PRA increased after transplant nephrectomy.New HLA allosensitization sites were found,which may be useful in HLA matching.