Aim To determine the effects of Qiliqian-gxin injected into lateral ventricle on Cardiac function, angiotensin Ⅱ( Ang Ⅱ) , angiotensin converting en-zyme(ACE), angiotensin type 1 receptor(AT1R) and the sympathetic nervous system in the hypothalamic pa-raventricular nucleus of rats with chronic heart failure. Methods Rat model of heart failure was prepared by acute myocardial infarction that was induced by ligation of the left anterior descending coronary artery. Four weeks after heart failure, Qiliqiangxin and Losartan were continuously administered via a syringe pump in-jector connected to lateral ventricle. After four weeks, echocardiogram was used to evaluate the cardiac func-tion and HE was used to observe myocardial tissue morphology, and enzyme linked immunosorbent assay was used to measure plasma norepinephrine( NE) , ser-um NT-proBNP and Ang Ⅱ in the paraventricular nu-cleus. The expression of ACE and AT1 R at mRNA and protein levels in the paraventricular nucleus was deter-mined by Real-time PCR and Western blot, and the RSNA was measured by PowerLab in anesthetized rats. Results Compared with the sham control, the cardiac function was significantly lower while the AngII, ACE, AT1 R expression in the paraventricular nucleus and RSNA were significantly increased in rats with heart failure. Compared with heart failure control, Qiliqian-gxin and Losartan decreased the RSNA and the AngII, ACE, AT1 R expression in the paraventricular nucleus. Conclusion Giving traditional Chinese medicine to the lateral ventricles can decrease the activation of the RAS system, reduce the renal sympathetic nerve activi-ty and improve cardiac function.

Two p-phenylenediamine (p-PD)-imprinted polymers, P (MAA) and P (AA), were synthesized using methacrylic acid (MAA) and acrylamide(AA) as functional monomer, respectively, in order to prepare molecular recognition material with high selectivity for p-PD and explore the feasibility of methods such as molecular spectrometry and computational approach of quantum chemistry for the selection of functional mono mer with high imprinting efficiency.The molecular recognition properties of the imprinted polymers were evaluated by high performance liquid chromatography.The results indicated that P(AA) exhibited no imprint ing effect for p-PD, while P(MAA) can bind p-PD selectively(k' =3.57), which showed remarkable imprint ing effect (IF=2.95), and p-PD and its analogues o-phenylenediamine and p-aminobenzoic acid can almost realize baseline separation on P (MAA) column in the mobile phase of methanol.Furthermore, we made a comparative study on the interaction of p-PD with MAA and AA by spectroscopic techniques such as UV and fluorometry as well as HF/6-31G~* computational approach.The results demonstrated that the complex of p-PD-MAA was more stable than that of p-PD-AA, which can give a good explanation for the molecular recog nition properties of P (MAA) and P (AA).The study indicated that both molecular spectrometry (UV and fluorometry)and computational approach of quantum chemistry can be employed as efficient means for the selection of efficient functional monomer.The results showed that fluorometry is sensitive and convenient for the choice of functional monomer if the template molecule is fluorescent.

Aim To investigate the treatment effect of Tongxinluo(TXL)combined peripheral blood derived mesenchymal stem cells(PB-MSCs)transplantation on angiogenesis of HIF-1 /VEGF pathway and miR-21 0 in diabetic foot(DF)rats.Methods Seventy male Spra-gue-Dawley rats were fed with high-fat diet and given
combined streptozotocin injection to build diabetic rat model.The femoral artery and vein of right lower ex-tremity of modeled rats were ligatured to induce DF model.The normal rats in the same batch copied is-chemia model served as control group.The modeled rats were divided into 6 groups :model group ,TXL
group,Cilostazol group,PB-MSCs group,TXL com-bined PB-MSCs(T-MSCs)group and Cilostazol com-bined PB-MSCs(C-MSCs)group.After the treatment for 28 days,animal gastrocnemius muscle and serum were taken for detection,in which serum was used to detect VEGF-A and HIF-1 α by ELISA,and muscle was used to detect VEGF-A and VEGF-R2 by Western blot,and VEGF-A by RT-PCR.Results Compared with the control group,the expressions of VEGF-A, HIF-1 α,VEGF-R2 and miR-21 0 in the model group were significantly reduced(P <0.01 ).Compared with the model group,the expressions of VEGF-A,VEGF-
R2 and miR-21 0 in each treatment group were signifi-cantly increased(P <0.01 ),where the expressions of VEGF-A,HIF-1 α,VEGF-R2 and miR-21 0 in the T-MSCs were higher than these in the TXL,cilostazol and PB-MSCs group(P <0.05,P <0.01 ).Conclusion TXL combined with PB-MSCs transplantation may be regulated by HIF-1 /VEGF pathway and miR-21 0,pro-moting endothelial cell proliferation,differentiation, and promoting the formation of new blood vessels.

Bone marrow mesenchymal stem cells (BMSCs),which can be isolated from organs and tissues,are cell populations with high degree of plasticity,similar to hematopoietic stem cells in bone marrow,and can be in vitro cultured,induced and amplified.BMSCs are increasingly used in gene therapy,cell therapy and tissue engineering.BMSCs have been currently studied in a number of autologous transplantations,while not all BMSCs are suitable for autologous transplantation.BMSCs exhibit biological aging gradually when cultured in vitro.Biological aging can be divided into age-induced senescence and long-term passage induced senescence.Aging BMSCs demonstrate changes in biological behaviour which reduces the success rate of autologous transplantation.In this review,biological aging BMSCs are discussed in order to provide help for the transplantation of BMSCs.

BACKGROUND:The microRNAs are involved in regulation of stem cel proliferation, differentiation and aging. To study the effect of Let-7c, a member of Let-7, on the neural differentiation of bone marrow mesenchymal stem cels provides new ideas for stem cel therapy. OBJECTIVE: To investigate the role of Let-7c in the neural differentiation of bone marrow mesenchymal stem cels. METHODS: The lentiviral vectors of Let-7c-up and Let-7c-inhibition were constructed and transfected into rat bone marrow mesenchymal stem cels. Optimal multiplicity of infection was screened. The cels were divided into non-transfected group, negative control group (transfected with empty virus), transfected enhancement group (transfected with LV-rno-Let-7c-up), transfected inhibition group (transfected with LV-rno-Let-7c-5p-inhibition). Bone marrow mesenchymal stem cels were treated with fasudil as an inducer for triggering the cels to differentiate into neurons. The fluorescence expressed by transfected cels was observed under inverted fluorescence microscope. The expression of neuron-specific markers, neuron-specific enolase and microtubule-associated protein 2, were measured by immunocytochemical method. The mRNA expression of microtubule-associated protein 2 was detected by RT-PCR. The cel viability was determined by MTT method. RESULTS AND CONCLUSION:Under the inverted fluorescence microscope, the cels were successfuly transfected with LV-rno-Let-7c-up and LV-rno-Let-7c-5p-inhibition. Fasudil induced bone marrow mesenchymal stem cels to differentiate into neurons. The transfection efficiency and expression levels of neuron-specific enolase and microtubule-associated protein 2 in the transfected enhancement group were significantly higher than those in the negative control group (P < 0.05), while in the transfected inhibition group, they were lower than those in the negative control group (P < 0.05). These findings indicate that the differentiation percentage of bone marrow mesenchymal stem cels is increased by fasudil after transfection with LV-rno-Let-7c-up, and Let-7c may promote the differentiation of bone marrow mesenchymal stem cels into neurons.

Aim To observe the effect of“Ying-nutrient and Wei-defense unblocking collaterals prescription”on the carotid sympathetic nerve in early atherosclero-sis. Methods The rabbits were randomly divided into 6 groups:Control group, high-fat group, Complex mod-el group, GSTL-H group, GSTL-L group and ATO group. The control group was fed with common food-stuffs, High-fat group with high-fat diet to build early atherosclerosis model, and all the other groups with high-fat diet combined with carotid artery cannula to build early atherosclerosis carotid artery injury rabbit model. All groups were given corresponding medicines intragastrically once a day for 4 weeks. The GSTL-H and GSTL-L group was given Guishaotongluo ultrafine powder 0. 6 g·kg-1·d-1, 0. 3 g·kg-1·d-1, and the ATO group was given suspension of atorvastain so-lution 2. 5 mg · kg-1 · d-1 . Biochemical method was used to detect blood lipid change. HE staining was ap-plied to observe the pathological morphology of intima-media, The content of NE in the carotid arterial was detected by ELISA. The immunohistochemical staining was used to detect the protein expression of NGF, TH around the carotid arterial. The expression of NGF, TH in the carotid artery adventitia was detected by Western blot. Results Compared with normal group, the levels of TC,TG and LDL-C in high-fat group and complex model group were significantly increased( P<0. 01). The degree of the intimal hyperplasia,the con-tent of NE and sympathetic density ( NGF, TH protein expression and distribution ) of the cartoid artery in complex model group were heavier compared with those in high-fat group; the lipid levels, degree of the inti-mal hyperplasia and sympathetic density ( NGF, TH protein expression and distribution ) in the GSTL-H, GSTL-L group were milder in varying degrees compa-ring with those in the complex model group(P<0. 05, P<0. 01 ) . Conclusion “Ying-nutrient and Wei-de-fense unblocking collaterals prescription” can reduce rabbit carotid atherosclerosis, and the mechanism may be related to regulating the sympathetic nerve of arterial wall.

[ ABSTRACT] AIM:To investigate the transient outward potassium channel protein expression in paraventricular nucleus (PVN) and its contribution to renal sympathetic nerve activity (RSNA) in rats with chronic heart failure (CHF). METHODS:A rat model of CHF was prepared by acute myocardial infarction that was induced by ligation of the left ante -rior descending coronary artery .Four weeks after heart failure , echocardiogram was applied to identify the CHF model and plasma norepinephrine (NE), serum NH2-terminal pro-brain natriuretic peptide (NT-proBNP) were detected by ELISA. The expression of ransient outward potassium channel proteins Kv 4.2 and Kv4.3 at mRNA and protein levels was deter-mined by real-time PCR and Western blot .The mean arterial pressure ( MAP) , heart rate ( HR) and RSNA were measured in anesthetized rats with PVN microinjection of potassium channel blockers 4-AP.RESULTS:In CHF group , the rat car-diac function and Kv4.2 and Kv4.3 expression in PVN were obviously lower while plasma NE and serum NT-proBNP were obviously higher than those in sham group .Microinjection of 4-AP into PVN induced an increase in MAP , HR and RSNA in both sham and CHF rats , while the CHF rats exhibited smaller responses to 4-AP than sham-operated rats .CONCLU-SION:Downregulation of Kv4.2 and Kv4.3 expression in the PVN may be a potential mechanism for sympathoexciation in the rats with chronic heart failure .

BACKGROUND:There is no clear understanding about the effect of let-7f and interleukin-6 (IL-6) on the proliferation of bone marrow mesenchymal stem cels and their relationship.
OBJECTIVE: To explore the effects of expression levels of let-7f and IL-6 on the proliferation of bone marrow mesenchymal stem cels and their relationship.
METHODS:(1) LV-rno-let-7f-up and LV-rno-let-7f-down were constructed and transfected into bone marrow mesenchymal stem cels of Sprague-Dawley rats, respectively. Then, there were four groups in the study: transfection upregulation group transfected with LV-rno-let-7f-up), transfection inhibition group (transfected with LV-rno-let-7f-down), negative control group (transfected with FU-RNAi-NC-LV), and untransfected group. The expression level of let-7f in each group was detected by qRT-PCR. The proliferation ability of cels and expression levels of IL-6 when let-7f expression was at different levels were detected by MTT, flow cytometry and ELISA. The expression of Cyclin D1 at mRNA and protein levels was detected by qRT-PCR and western blot, respectively. (2) To predict the potential target gene of let-7f, the wild-type/mutant IL-6 3’UTR reporter gene vectors were constructed, and cotransfected with let-7f/let-7f inhibitor respectively into the 293T cels to measure the luciferase.
RESULTS AND CONCLUSION: Compared with the negative control group, the proliferative and cloning capacities of cels in the transfection upregulation group were higher; the number of cels was significantly decreased at G1 stage and increased at S stage, and the apoptotic cels were reduced in number (P < 0.05). However, the transfection inhibition group had opposite results. The expression level of IL-6 in the transfection upregulation group was lower than that in the untransfected group and negative control group (P < 0.05); while in the transfection inhibition group, the expression level of IL-6 was significantly increased (P < 0.05). The expression of Cyclin D1 at mRNA and protein levels was up-regulated in transfection upregulation group (P < 0.05) and down-regulated in the transfection inhibition group (P < 0.05), but there was no significant difference between the negative control group and untransfected group (P > 0.05). Luciferase activity of cels transfected with wide-type IL-6 3’UTR and let-7f was significantly reduced (P < 0.05). These findings indicate that up-regulation of let-7f can promote the proliferative and cloning capacities of bone marrow mesenchymal stem cels and reduce cel apoptosis, but downrelation of let-7f exhibits an inhibitory effect. Overexpression of IL-6 can suppress the proliferation of bone marrow mesenchymal stem cels, which is considered to be a target gene of let-7f, and let-7f may suppress the expression of IL-6 to promote the cel proliferation.

BACKGROUND:MicroRNA plays an important role in the process of growth and aging of living body. To know the role of let-7d in inducing bone marrow mesenchymal stem celldifferentiation into neurons can promote the stem celltransplantation. OBJECTIVE:To investigate the role of let-7d in inducing bone marrow mesenchymal stem celldifferentiation into neurons. METHODS:(1) The lentiviral vector of let-7d was constructed and transfected into rat bone marrow mesenchymal stem cells. The cells were divided into non-transfected group, negative control group (transfected with FU-RNAi-NC-LV), transfected enhancement group (transfected with let-7d-LV), transfected inhibition group ( transfected with let-7d-inhibition-LV). (2) Rat bone marrow mesenchymal stem cells were treated with fasudil as an inducer for triggering the cells to differentiate into neurons. The expression of neuron-specific markers, neuron-specific enolase and microtubule-associated protein 2, were measured by immunocytochemical method. The mRNA expression of microtubule-associated protein 2 was detected by RT-PCR. The viability of bone marrow mesenchymal stem cells was determined by MTT method. RESULTS AND CONCLUSION:Under inverted fluorescence microscope, the cells were successful y transfected with let-7d. Fasudil induced bone marrow mesenchymal stem cells to differentiate into neurons. The transfection efficiency and expression levels of neuron-specific enolase and microtubule-associated protein 2 in transfected enhancement group were higher than those in the negative control group (P<0.05);while in the inhibition group, they were lower than those in the negative control group (P<0.05). These findings indicate that let-7d can promote the differentiation of bone marrow mesenchymal stem cells into neurons induced by fasudil, and by control ing the expression of let-7d we can influence the differentiation efficiency from bone marrow mesenchymal stem cells to neurons.

Objective To analyze the differential expressions ofmicroRNAs in patients with methylmalonic acidemia (MMA) and explore the expression rule ofmicroRNAs in MMA pathogenesis primarily to find the new biomarkers and therapeutic evaluation index of MMA represented by microRNAs.Methods Ten patients,admitted to our hospitals and diagnosed as having delayed onset vitamin B12 valid MMA from August 2011 to June 2013,were chosen as experimental group (MMA group);and their 8 healthy relatives were chosen as negative control group (NC group).Plasma microRNAs were routinely extracted and filed,and samples quality was evaluated with real-time quantitative PCR;microarray gene chip was employed to detect the expression levels ofmicroRNAs;R software of prediction analysis ofmicroarrays (PAM) was used to filter differentially expressed miRNAs.Results The results of GC/MS showed that the urine methylmalonic acid zoom ratio of MMA group was significantly higher than that of NC group.And the urine methylmalonic acid zoom ratio of MMA group had remarkable difference between before and after treatment.Real-time PCR showed the RNAs extracted from plasma samples conformed to the requirement of experiment and could be delivered to downstream chip experiment.The chip statistical analysis suggested that there were many micrornas enjoying significant differences between MMA group and NC group (P＜0.05),and differences in MMA group before and after treatment (P＜0.05).The resluts of R software of PAM indicated that mir-483 and mir-144 were strongly raised in MMA group as compared with those in NC group (P＜0.05),while mir-151,mir-30 and mir-146 were remarkably reduced in MMA group as compared with those in NC group (P＜0.05).Conclusion There are several abnormal expressions of plasma circulation microRNAs in patients with methylmalonic acidemia;the plasma circulation microRNAs might be biomarkers of methylmalonic acidemia.