Object To establish an effective system for Agrobacterium mediated genetic transformation of Poncirus trifoliata Raf Methods The explants used for transformation were the epicotyls from P trifoliata; the Agrobacterium tumefaciens strain was EHA101, provided with the vector plasmid pGA482GG; the coat protein gene (CTV cp gene) was introduced into the transformation plasmid The transformation was proved by PCR and histochemical GUS assay Results In trans for mation of P tri foliata, 20 days epicotyls were suitable for transformation Shoot regeneration frequency was high when cocultivation time was 2-3 days The presence of acetosyringone during cocultivation could enhance the efficiency of transformation Histochemical GUS assay showed that 70 0% of the resistant plants were GUS positive Extra gene was proved to be transformed into P trifoliata plant by PCR analysis Conclusion An effective genetic transformation system has been established for P trifoliata This system will be useful for resistant breeding of P trifoliata

The herpes simplex virus type 1 (HSV-1) VP22, is one of the most abundant HSV-1 tegument proteins with an average stoichiometry of 2 400 copies per virion and conserved among alphaherpesvirinae. Many functions are attributed to VP22, including nuclear localization, chromatin binding, microtubule binding, induction of microtubule reorganization, intercellular transport, interaction with cellular proteins, such as template activating factor I (TAF-I) and nonmuscle myosin II A (NMIIA), and viral proteins including tegument protein VP16, pUS9 and pUL46, glycoprotein E (gE) and gD. Recently, many novel functions performed by the HSV-1 VP22 protein have been shown, including promotion of protein synthesis at late times in infection, accumulation of a subset of viral mRNAs at early times in infection and possible transcriptional regulation function.

Objective To construct a new recombinant immunotoxin expression vector fused with a murine interleukin18(IL18) gene and a truncated pseudomonas exotoxin (PE38) gene, and examine the expression of IL-18-PE38 fusion protein in Escherichia coli (E. coli). Method Murine IL-18 (mIL-18) cDNA was cloned from murine liver tissue through reverse transcription-polymerase chain reaction (RT-PCR). The mIL-18 cDNA was ligased with a PE38 gene carried by PRKL expression vector through T4 DNA ligase and constructed into fusion protein expression plasmid PRKL-IL18-PE38. The recombinant vector was identified by restriction endonucleases digestion, PCR and DNA sequencing. After transformed into E.coli BL21 and induced by IPTG, the expressed product was obtained and the molecular weight and specificity were determined by SDS-PAGE and Western-blotting. Result The new recombinant immunotoxin expression vector was constructed successfully. DNA sequencing revealed that the mIL-18 and PE38 gene were consistent with NCBI Gene Bank. The IL-18-PE38 fusion protein was expressed in E.coli BL21, and Western-blotting analysis indicated that the molecular weight of the expression product is about 56 kDa, and could react with the specific antibody against mIL-18. Conclusion IL-18-PE38 recombinant immunotoxin expression vector will provide the basis for study on the targeted cytotoxic activity to Th1 cells and may have some potential value in the treatment of Th1 cell-mediated autoimmune diseases.

[Abstratc] Objective The function of cIAP1 in the progression of ovarian cancer has not been clarified . This study is to explore the involvement of cIAP 1 in regulating biological behaviors of ovarian cancer cells by u-sing RNA interference(RNAi)technology.Mte hods The short hairpin RNA plasmid targeting cIAP1 was con-structed and transfected into Skov 3 cells.The levels of cIAP1 mRNA and protein were investigated by RT -PCR and Western Blot respectively .MTT assay and flow cytometry were used to evaluate cell proliferation and apopto-sis.R esults The rate of cIAP1 transfection was 74.7%performed by flow cytometric analysis .cIAP1 expression was significantly down -regulated at both mRNA and protein levels ,which resulted in a decrease of cell prolifera-tion and invasion capability in vitro .Conclusion This study implies that cIAP 1 might play an important role in the progression of ovarian cancer ,and it could be a potential target for therapeutic anti -cancer drugs .

Objective:To explore the correlation of nurses ’ organizational commitment and their professional identity status.Methods:Using Chinese employee organizational commitment scale and nurses professional identity evaluation scale questionnaire survey was conducted in 400 cases of clinical nurses , nurse compare different characteristics of organi-zational commitment and professional identity level , and USES the Pearson correlation analysis to investigate the correla-tion of both.ResultsNurses organizational commitment and professional identity total score , respectively (2.16-0.45) and (3.49 +0.84);Different cultural degree , job title, working years and the way of hiring nurses ’ professional identi-ty score comparison, the difference was statistically significant (P <0.05);Different ways of title, working years, and hire nurses’ organizational commitment score comparison , the difference was statistically significant (P <0.05);Pear-son correlation analysis showed that organizational commitment and the scores of each dimension career cognitive apprais -al, professional social skills , professional setback coping , social support and total scores of professional identity were sig -nificantly positive correlation (P <0.05).Conclusions:The nurse was significantly positively related to organizational commitment and their professional identity , and both have big room to improve .

Objective:To analyze the chemical constituents of the ether extracts from the fruits of 3 species of Chaenomeles . Methods: The chemical constituents were analyzed by GC MS. Results: Sixty one constituents were firstly isolated and identified from the fruits of 3 species of Chaenomeles . Forty compounds in Chaenomeles lagenaria , thirty four compounds in Xuan mugua ( C.lagenaria from Xuanzhou) and 23 compounds in C.sinensis were isolated and identified respectively. There were 10 compounds in all 3 species of Chaenomeles , but their contents were very much different. They were acetic acid, benzaldehyde, n caproic acid, 14 methyl pentadecanoic methyl ester, glycerin, benzoic acid, 6,9 octadecadienoic methyl ester, squalane, stearic acid and oleic eicosyl ester. Conclusion: The composition and content vary with different species and district of Chaenomeles . The result may be useful for the evaluation of Chaenomeles fruits quality. [

The rennin angiotensin system has been recognized for many years as a key regulator in systemic blood pressure and the metabolism of water and salt. Angiotensin converting enzyme 2(ACE2) is the first human homolog of angiotensin converting enzyme which efficiently hydrolyze the angiotensin Ⅱ to vasodilator angiotensin 1-7.ACE2 is now implicated in cardiovascular,renal, lung disease and serves as a receptor for Severe Acute Respiratory Syndrome coronavirus.

Objective: To observe the clinical effect of Tongfu granules(通腑颗粒) on the gastro- intestinal dysfunction in patients with multiple organ dysfunction syndrome(MODS).Methods: The trial was prospective,multi-centric and clinically controlled.One hundred and forty patients with MODS who had been selected were randomly divided into two groups: mosapride citrate group and Tongfu granules group.Respectively at 0 hour,the 48 th hour,the 7 th day or before death, the following scoring systems were calculated: the intestinal dysfunction score,acute physiology and chronic health evaluationⅡ((APACHEⅡ)) score and Marshall score.The duration of mechanical ventilation,hospitalization in intensive care unit(ICU) and the prognosis within 28 days were recorded.Results: After treatments,the intestinal dysfunction score,(APACHEⅡ) score and Marshall score of all patients decreased,at the same time,the therapeutic effects of Tongfu granules group were more significant than those in mosapride citrate group(P20 scores.The mortality was elevated with the increased number of dysfunction organs.Conclusion: Tongfu granules can ameliorate the severity of the disease situation andimprove the prognosis of patients with MODS.

OBJECTIVETo gain a primary understanding of the prevalence of 21-hydroxylase deficiency(21-OHD) heterozygote (carrier) among androgen excess women of Chinese Han nationality, compare the molecular genetic changes therein revealed with the results of adrenocorticotropic hormone (ACTH) stimulating test, and assess the carriers' phenotype-genotype correlation.

METHODSEighty-two androgen excess cases and 14 healthy women underwent ACTH stimulating test during the follicular phase. Molecular genetic analysis of CYP21 for 9 common mutations was performed with the method of amplification-created restriction sites.

RESULTSIn androgen excess group, the basal level of F0 (P<0.01), as well as basal 17-OHP0 and the ACTH stimulated concentrations of 17-OHP60 were much higher than controls (P<0.01), and there was no obvious discrepancy in F60 (P>0.05). The net increase of 17-OHP and the ratio of net increase of 17-OHP to net increase of F were also higher than controls (P<0.01). No CYP21 gene mutations were found in control group. Four patients of the androgen excess group were identified as heterozygous carriers of CYP21 mutations. The ACTH stimulating test results from gene normal patients and from carriers overlapped to a certain extent.

CONCLUSIONAmong 82 patients of Chinese Han nationality androgen excess women, 4.9% were 21-OHD heterozygous. The response of serum 17-OHP is not useful for predicting CYP21 gene mutation carrier status. Genotyping is the most reliable method to detect carrier.

Adolescent ; Adrenal Hyperplasia, Congenital ; diagnosis ; ethnology ; genetics ; Adrenocorticotropic Hormone ; Adult ; Asian Continental Ancestry Group ; genetics ; Child ; China ; Female ; Genetic Carrier Screening ; methods ; Genotype ; Humans ; Mutation ; Polymerase Chain Reaction ; Steroid 21-Hydroxylase ; genetics ; metabolism

Objective To study the protective effects of the total bakkenolides from P .tricholobus on high altitude hy-poxia .Method Normobaric hypoxia model and acute hypobaric hypoxia model in mice ,hypobaric hypoxia model in rats were established for this study .Survival time and survival rate of mice were recorded .The level of blood sugar and glycogen ,adeno-sine triphosphate (ATP) ,lactic acid (LD) ,lactic dehydrogenase (LDH) were detected in different organs of rats .Results The total bakkenolides significantly prolonged the survival time of mice in normobaric hypoxia model and reduced the death rate of mice in acute hypobaric hypoxia model .The total bakkenolides suppressed blood sugar level in rats and increased the glyco-gen level in rat liver ,skeletal muscle and myocardium .It also elevated the ATP content in rat brain ,liver ,skeletal muscle and myocardium .Meanwhile ,the content of LD in plasma ,skeletal muscle ,myocardium and LDH level in myocardium were re-duced .Conclusion The total bakkenolides from P .tricholobus have protective effect on normobaric hypoxia model and acute hypobaric hypoxia model in mice as well as hypobaric hypoxia model in rats .Its anti-hypoxia efficacy at high altitude may relate to the increased blood sugar ,glycogen ,and ATP level and reduced LD ,LDH level in major organs .