BACKGROUND: Insulin like growth factor 2 (IGF2) is an important embryonic mitosis growth promoting factor, whichplays a critical role in the process of maintaining normal cell growth. The gene expression of IGF2 is under epigeneticregulation in the way of genomic imprinting. Imprinting loss of IGF2 is likely to be associated with the abnormaldevelopment of the individual and tumorigenesis.OBJECTIVE: To investigate the effect of imprinting loss of IGF2 gene on the differentiation of mouse embryonic stemcells into islet-like cells.METHODS: Two kinds of mouse embryonic stem cells (SF1-G and SF1-1) were induced to differentiate into islet-likecells in vitro. The expression of Insulin gene was detected by real-time PCR and cell immunofluorescence. Theexpression of IGF2 was detected by the polymerase chain reaction-restriction fragment length polymorphism in the cellsbefore and after induced differentiation. The level of insulin released at terminal differentiation stage was tested by insulinrelease assay in vitro.RESULTS AND CONCLUSION: (1) There was no change in the imprinting state of the two kinds of mouse embryonicstem cells with normal and imprinted IGF-2 gene before and after differentiation. (2) In the induced cells, the expressionlevel of insulin in the SF1-1 group was higher than that in the SF1-G group, although there was no significant differencein the insulin release between the two kinds of cells. (3)The imprinting loss of IGF-2 gene may be related to theup-regulation of insulin mRNA expression in terminal cells during induced differentiation.

Object To determine the content of total astragalan in FUZHENG YILIU GRANULES Methods The determination was performed by phenol-sulfuric acid method Results The content of total astragalan in FUZHENG YILIU GRANULES was 3 0%, the linearity of this method was quite satisfactory (r= 0 998 8), and the average recovery rate was 97 95%, RSD=1 85% Conclusion This method was simple, accurate and reproducible, and available for quality control of the GRANULES

Objective To investigate the expression of phosphorylated protein kinase B1 (pAkt1) and human telomerase reverse transcriptase (hTERT) in ovarian epithelial carcinoma (EOC) and its relationship with prognosis, and to analyze the correlation between pAkt1 and hTERT expression. Methods 92 patients with EOC in Tangshan Gongren Hospital from January 2012 to December 2016 were selected. The expressions of hTERT and pAkt1 proteins were detected by immunohistochemistry. The relationship between the expressions of pAkt1 and hTERT proteins and their relationship with clinical prognosis were analyzed. Results Of the 92 patients with EOC, 68 cases (73.9 %) had positive expression of pAkt and 52 cases (56.5 %) had positive expression of hTERT. There was no significant correlation between expressions of pAkt1 and hTERT proteins in EOC (r= 0.284, P= 0.633). The expressions of pAkt1 and hTERT proteins were not related with age, tumor pathology type and the International Federation of Gynecology and Obstetrics (FIGO) staging (all P> 0.05), but had significant association with tumor pathology differentiation (χ2= 2.694, P= 0.005; χ2=2.284, P=0.018). The disease-free survival of patients with both pAkt1 and hTERT positive was shorter than that of the other groups (P= 0.013). Conclusion The prognosis of EOC patients with high expression of pAkt1 and hTERT proteins is poor.

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AIM: To express chimeric toxin Stx2a’-LHRH a nd to investigate the cytotoxic activity of recombinant toxin Stx2a’-LHRH to huma n carcinoma cells.METHODS: Stx2a’-LHRH sequences that added the res tri ction endonucleases NcoⅠ and EcoRⅠ at the 5' and 3 ends were amplified by PCR a nd digested with appropriate restriction enzymes. The digested fragment was subc loned into the vector obtatined by digestion of plasmid pET-28a(+) with NcoⅠ an d EcoRⅠ. E. coli BL21 (DE3) cells were transformed with plasmids of interst and cultured in LB medium containing ampicillin. Expression of the recombinant protein was induced by the addition of isopropylthio-?-D-galactoside (IPTG). T h e cytotoxity of Stx2a’-LHRH to Hep-2 cells was observed under the microscop y. RESULTS: Recombitant plasmid pET-SL was constructed successfu lly and the clones expressing pET-SL stablely were obtained. A special electroph oretic band in SDS-PAGE (a glycoprotein of 28kD) was noted. Stx2a’-LHRH killed He lp-2 cells clearly. CONCLUSION: In this study, construction of c himeric toxin Stx2a’-LHRH and its expression were described. Moreover, it has o bvious cytotoxity to Hep-2 cell. These finding could open up new vistas in the s tudy of targeted durgs.

Objective To evaluate the expressions and prognostic significance of phosphorylation of protein kinase B1 (pAkt1) and human telomere reverse transcriptase (hTERT) in epithelial ovarian carcinoma.Methods The expressions of pAkt1 and hTERT were examined by immunohistochemical SP method in 72 cases of epithelial ovarian carcinoma and 10 normal endometrial tissues to analyze the correlation between pAkt1 and hTERT and their relationships with clinic-pathological features and prognosis.Results The positive expression rate of pAkt1 in epithelial ovarian carcinoma was 73.6％,and the positive expression rate of hTERT was 56.9％,compared with normal ovarian tissues,with statistical significance (x2 =20.814,P ＜ 0.001;x2 =11.3g9,P ＜ 0.001).The expression of pAkt1 was not associated with hTERT expression (r =0.075,P =0.532) in epithelial ovarian carcinoma.The expressions of pAkt1 and hTERT in epithelial ovarian carcinoma were correlated with tumor differentiation status (x2=6.475,P =0.011;x2 =1.370,P =0.001).The disease-free survival of patients with pAkt1 and hTERT positive coexpression was significantly shorter than others (8.585 months vs.11.227 months,x2 =4.361,P =0.037),but there was no significant difference in overall survival (11.107 months:11.695 months,x2 =0.394,P =0.530).Conclusion pAkt1 and hTERT are highly expressed in epithelial ovarian carcinoma,and high coexpression of pAkt1 and hTERT indicates poor prognosis in epithelial ovarian carcinoma.

Aim To explore the effect of receptor component protein(RCP)in the signal transduction of vascular smooth muscle cell(VSMC)proliferation induced by static pressure.Methods The mouse-derived vascular smooth muscle cell line(A10VSMC)was employed in the experiment.Cells were exposed to static pressure,and MTT assay was used to detect the cell viability.Western blot was used to determine the expressions of PCNA,RCP and p-Akt,RCP mRNA was tested by RT-PCR,and co-immunoprecipitation was used to test the interaction between RCP and G proteins.Results The cell viability,expressions of PCNA and RCP increased with the elevation of static pressure and reached their peaks at 120 mmHg,and after 6 hours they got a plateau.The static pressure significantly increased the level of p-Akt,meanwhile,the binding of RCP and Gαs significantly decreased.However,the binding of RCP and Gβ increased in response to static pressure after stimuli of static pressure,but Gγ was obscure.Conclusion Static pressure can induce VSMC proliferation and expression of RCP,which may involve G protein signal transduction model.

Adolescent ; Adult ; Emergencies ; Female ; Hand Injuries ; surgery ; Humans ; Male ; Middle Aged ; Surgical Flaps

Objective To investigate outcome and cognitive changes of amnestic mild cognitive impairment (aMCI) in a follow-up study. Methods A cross-sectional and longitudinal parallel cohort study design was conducted among 109 aMCI patients and 104 matched normal controls. Multi-dimension neuropsychologic tests were used to extensively assess the cognitive function. Results The scores of neuropsychologic tests in aMCI patients were significantly lower than those in the normal controls( all P＜0.01 ) ,with the largest impairment on 20minutes delayed recall of the auditory verbal memory test ( AVMT), which reflects episodic memory ( aMCI pa-tients :2.50 ± 1.48, normal controls :7.85 ± 1.59, Z = - 12.697, P ＜ 0.01 ); AD was diagnosed in 15 of the 69aMCI patients with a prevalence rate of 22% ,but none was converted to AD in the normal controls. The cognitivechanges of performance in AVMT, CDT, MMSE of the patients in aMCI group (( 3.77 ± 60.83 )%, (6.89 ±28.24) %, (6.13 ± 16.89) % respectively) were significantly poorer than those of the controls group(( - 10.75 ±27.46) %, ( - 5.23 ± 14.05 ) %, ( - 1.11 ± 8.26 ) % respectively) ( all P ＜ 0.05 ). At baseline, demented aMCIperformed poorer in AVMT, CFT, TMT, SDMT, CDT, MMSE when compared to stable. During the follow-up, demented aMCI groups performed significantly poorer than did stable subjects in AVMT, CFT, DST, VFT, SDMT,MMSE ( all P ＜ 0.05 ). Conclusion aMCI is a prodromal period of AD and characterized by episodic memory impairment. The neuropsychologic test is a predictive factor for aMCI to develop AD.

Adult ; Case-Control Studies ; China ; epidemiology ; Female ; Humans ; Iodine ; urine ; Male ; Middle Aged ; Thyroid Diseases ; epidemiology ; urine