Ob jective To construct and screen out the RPL 23-siRNA interference fragments ,providing the basis for the following experiments about the correlation with RPL 23 and gastric cancer .Methods The RPL23-siRNA,synthesized chemically through lipofection ,were selected from three target sequences by RNA in-terference and detected by real -time PCR and Western blot .Results Compared with normal cell group and RPL23 control group ,the mRNA and protein expression of RPL 23 in the other 3 interference groups were signifi-cantly decreased(P<0.01).Multiple comparisons showed that the interference efficiency of RPL 23 -siRNA1 group was significantly higher than that of RPL 23-siRNA2 group and RPL23-siRNA3 group(P<0.01).Con-clusion The RPL23-siRNA interference fragment can be successfully constructed and screened out ,which pro-vides the basis for the following experiments .

[ABSTRACT]AIM:ToinvestigatetheeffectofimmunosuppressantFK506onserumglucoseinratsandtoex-plore its mechanism .METHODS: Sprague-Dawley rats ( n =12 ) were randomly divided into drug group and normal group.The rats in drug group were intraperitoneally injected with FK 506 at dose of 1 mg· kg-1 · d-1 and the rats in nor-mal group received saline (1 mL· kg-1 · d-1 , ip) for 14 d.The fasting weight and fasting glucose were regularly meas-ured every 2 d.Visceral fat was isolated from the rats at the end of experiment .The mRNA expression of adiponectin , lep-tin, visfatin, resistin, retinol-binding protein 4 ( RBP4) and peroxisome proliferator-activated receptors γ( PPAR-γ) was determined by real-time fluorescence quantitative PCR .The protein expression of PPAR-γand adiponectin was measured by Western blotting .RESULTS:Compared with normal group , the concentration of fasting blood glucose in model group was significantly increased from the 10th day (P<0.05).At day 14, the fasting blood glucose of the model group increased from (5.10 ±0.62) mmol/L to (7.73 ±0.73) mmol/L.No significant change of blood glucose in normal group between the 10th day and the 14th day [from (4.66 ±0.32) mmol/L to (5.80 ±0.10) mmol/L] was observed.Compared with normal group , the mRNA expression of PPAR-γ, adiponectin and leptin in the adipose tissue of model group was signifi-cantly decreased ( P <0.01 ) , whereas the expression of visfatin , resistin and RBP4 was significantly increased ( P <0.05).Compared with normal group, the expression of PPAR-γand adiponectin in model group was decreased (P <0.01).CONCLUSION:FK506 may decrease the expression of PPAR-γto change the expression of adipocytokines and induce hyperglycemia in rats .

Objective:To explore the effect of adipose derived mesenchymal stem cells to regulate the differentiation of macrophage RAW264.7.Methods:First,we used RAW264.7 cells to simulate macrophage and induced them to M 1 macrophage with lipopolysaccharide ( LPS,1 μg/ml) .Then we cultured these RAW264.7 cells in culture mediums which were previously used to culture adipose derived mesenchymal stem cells to imitate the transplantation of ADMSC .Last,the mRNA relative expression of IL-10, IGF-1,Arg-1,TNF-α,FIZZ1,SPHK-1 was detected by real-time PCR.The protein expression of IL-12 p40,IL-27 Rα,IL-10 was detected by Western blot.Results:After been cultured in ADMSCCM and induced by LPS ,M1 markers (TNF-αmRNA,IL-12 p40;P<0.05) of the RAW264.7 cells declined while M2 markers (IGF-1 mRNA,IL-10 mRNA,IL-10;P<0.05) rose.Conclusion: ADMSC can secrete soluble cytokines to induce the RAW264.7 cell,which have been induced to the M1 macrophages,to differentiate towards M2 macrophages.

Objective In cases and events of mixed STR typing of aborted fetus,two methods for calculating paternity index(PI)of suspected biological fathers are proposed,which could be useful for theoretical reference for parental identification including mixed STR typing.Methods Depending on whether the fetal genotypes can be identified,the simple PI calculation method and the PI calculation method of deduced biological paternal genes when the fetal genotypes cannot be identified are proposed.Results The simple PI calculation method is to indentify the fetal genotypes first and then calculate according to the standard triplet.The PI calculation method of deduced biological paternal genes is to deduce all the possible genotypes of biological fathers conforming to Mendel's law(inference)without considering the ratio of peak height and peak area in mixed typing,and then calculate the parental index separately,taking the minimum value as the parental index of the locus.Conclusion When mixture ratio of fetus in the mixed typing of aborted tissue MR≥0.43,the accuracy of separation is very high and the simple PI calculation method can be accurate,so it is recommended.If 0.05≤MR＜0.43,it is suggested to use the calculation method of deduced biological paternal genes,which can avoid misjudgment of irrelevant persons to the greatest extent.If MR＜0.05,there's a high risk of fetal allele loss,we should not perform a paternity test on the mixed spot.Since the cumulative parental index calculated by deduced the biological paternal genes is usually lower than the value calculated by dividing the fetal genotype,the CPI may be lower than 10 000 when fewer loci are identified,and then more genetic markers should be detected.