This study was aimed to observe four kinds of kidney-tonification medicine, which were Epimedium, pso-ralen, Ligustrum lucidum, Polygonum with the active ingredient of icariin, psoralen, oleanolic acid, stilbene glucoside and their orthogonal compatibility. There were two kinds of non-kidney tonification medicine, which were Chuanx-iong and astragalus with the active ingredient of TMP and astragaloside. The observation was made on the regulatory role of rat bone marrow stem cells (BMSCs). A total of 65 SD rats were randomly divided into the normal control group, positive transformed control group, kidney-tonification compatibility group (including Group 1, Group 2, Group 3, Group 4, Group 5, Group 6, Group 7, Group 8, and Group 9), non-kidney tonification medicine control group (in-cluding TMP group and astragaloside group). Intragastric administration of medication was given to the kidney-tonifi-cation compatibility group and the non-kidney tonification medicine control group, once a day for 3 consecutive days. Intragastric administration of equal amount of normal saline was given to the normal control group and the posi-tive transformed control group. On the third day of intragastric administration, rats in each group were sacrificed. Serum containing medication was used in the culture of BMSCs for 6, 12, or 18 days. ELISA method was used to quantitatively detect the expression activity and content of BMP2 on the 6th, 12th, or 18th day, in order to evaluate the degree of bone cell differentiation degree. Real-time quantitative PCR method was used for detection of expression of Bmp2, Smad1, Smad4 mRNA in serum containing medication in the culture of BMSCs on the 18th day. The results showed that the kidney-tonification compatibility can improve the expression activity and content of BMP2 culture in vitro, with the peak on the 12th day. The kidney-tonification compatibility groups can upregulate expressions of Bmp2, Smad1, Smad4 mRNA. It was concluded that the active ingredient compatibility of kidney-tonification medicine can promote BMSCs. Its mechanism may be related to the upregulation of expression of Bmp2, Smad1, Smad4 mRNA, and the activity and content of Bmp2.

Objective To apply the virtual reality in balance function training for Parkinson's Disease (PD). Methods From October, 2013 to January, 2016, 31 patients with balance disorders after PD were divided into two groups:the treatment group (n=17) received virtual reality balancing games training, while the control group (n=14) received routine balance training. They were assessed with Berg Balance Scale (BBS), Timed Up and Go Test (TUGT), static fall index (SFI), limits of stability (LOS), dynamic fall index (DFI) and modified Barth-el Index (MBI) before and four weeks after treatment. The frequency of falls in the next six weeks was recorded. Results The scores of BBS, TUGT, SFI, LOS, DFI and MBI improved significantly in both groups after treatment (P<0.05), and improved more in scores of BBS (t=2.095), TUGT (t=-2.091), DFI (t=-2.182), LOS scores (t=2.202) and MBI (t=3.036) in the treatment group than in the control group (P<0.05). There was positive correlation between scores of BBS and MBI (r=0.899, P<0.001). The frequency of falls was less in the treatment group than in the control group (P<0.05). Conclusion Virtual reality balance training can improve the balance function for PD.

Objective:To investigate the change of expression of miR-7 in activated CD4+ T cells in vitro, and preliminary explore its possible significance. Methods: CD4+CD62L+T cells was purified from splenocytes of FVB mice by magnetic cell sorting system (MACS). After stimulation with anti-CD3/CD28 antibody,the relative expression of miR-7 was examined by Real-time PCR, and the expression level of CD69 molecular was analyzed by FACS. Furthermore,the relative expression of miR-7 in CD4+T cells was detected at different time points during stimulation. With the treatment of ERK inhibitor PD98059,change of miR-7 expression was de-termined by Real-time PCR. Meanwhile, the proliferation of CD4+T cells was examined by CCK-8 assay and the expression level of CD69 and CD62L molecular were analyzed by FACS. Finally,the expression of cytokines IL-6,IL-10,and IFN-γ were determined by Real-time PCR. Results:Compared with control group,the relative expression of miR-7 was increased significantly after stimulation with anti-CD3/CD28 antibody,as well as expression level of CD69 molecular was augmented(P<0. 05). In contrasted with 0 h and 24 h,the expression of miR-7 was significantly increased after 48 h and 72 h during stimulation(P<0. 05). Furthermore,the relative expression of miR-7 was significantly declined in CD4+T cells in ERK inhibitor PD98059 treatment group. Finally, the expression level of CD69 molecular,as well as cytokines IL-6, IL-10 and IFN-γ, were also decreased significantly ( P<0. 05 ) . Conclusion: The relative expression of miR-7 was significantly increased in activated CD4+T cells,closely related to ERK pathway,which provided an important foundation for successive research work on exploring the functional role of miR-7 in the CD4+T cells.

Objective:To investigate the influence of miR-7 knock down on the development of CD 4+SP cells in the thymus in mice,and preliminary explore its possible mechanism.Methods:The changes of volume ,weight and total cell counts of thymus in miR-7 knock down (miR-7KD) mice were observed compared with Wild-type(WT)mice;the pathological changes of thymus were observed by HE staining.FACS analysis was performed on the proportion ,as well as the expression level of CD44 and CD62L,of thymus CD4+single positive (SP) cells.Meanwhile,the proliferation percentage of CD4+SP cells was measured by Ki-67 staining.The apoptosis percentage of CD4+SP cells was analyzed by FACS.The changes on the transduction of ERK 1/2 pathways were determined by Western blot.Results:Compared with WT mice ,the size,weight and total cell number of thymus were marked reduced in miR-7KD mice( P<0.05 );moreover ,pathological change also was presented.The proportion and total cell number of thymus CD 4+SP cells were marked decreased ( P<0.05 ).Furthermore ,the expression level of CD 44 and proliferation percentage ,as well as apoptosis percentage ,of CD4+SP cells were obviously increased (P<0.05),however,the expression level of CD62L of CD4+SP cells were decreased (P<0.05). Finally,the level of total ERK1/2 and phosphor-ERK1/2 was decreased obviously ( P<0.05 ).Conclusion: miR-7 knock down can affect the development of CD 4+SP cells in the thymus , which might be closely related to the cell activation state and altered the transduction of ERK1/2 pathways.

This article was to elucidate that the needling depth is closely related to the meridian qi, disease location, disease nature and needled area based on the records of needling depth in Nei Jing (Canon of Internal Medicine). Moreover, different depths will produce different therapeutic efficacies;meanwhile, improper depth may lead to grave consequences.

AIM: To investigate the relationship between the intracellular expression of Th1 and Th2 type cytokines in placental lymphocytes and the pregnancy outcomes in NOD/SCID mice. METHODS: The resorption rate of embryos (RR) was compared between BALB/c and NOD/SCID mice. In addition, the expression rates of Th1 type (TNF-? and IL-2) and Th2 type (IL-10) cytokines were detected intracellularlly in placental lymphocytes by four-color flow cytometry. RESULTS: Although multiple-immunodeficiency was confirmed in the NOD/SCID, no significant difference was observed in RR between BALB/c and NOD/SCID mice. At the same time, a dramatically elevated level of CD8+IL-10+/CD8+ cell percentage was found at the feto-maternal interface in NOD/SCID?NOD/SCID, as compared with BALB/c?BALB/c mice, while no significant difference was observed for TNF-? and IL-2 expression in CD4+ and CD8+ cell subsets isolated from these mice. CONCLUSION: The spontaneous elevation of CD8+IL-10+/CD8+ cell percentage at the feto-maternal interface may be related to the roughly normal fertility in NOD/SCID mice.

Objective:To detect the effect of microRNA-7 ( miR-7 ) knockdown on pathology in murine acute lung injury ( ALI) model,and preliminarily explore its significance.Methods:Murine ALI model was performed by intraperitoneal injection of Li-popolysaccharide (LPS) (10 mg/kg) into miR-7KD mice and wild-type (wild type,WT) mice respectively.Then,the pathologic injury of lung tissue were observed by HE staining.And total cell count of bronchoalveolarlavage(BAL) was calculated.The relative expression of related cytokines in lung tissue was analyzed by Real-time PCR assay.Furthermore,the changes on proportion of innate immune cells (γδT cell and F4/80 macrophages cell) and adaptive immune cell ( CD4+T cell and CD8+T cell) were analyzed by FACS.Meanwhile, the expression of CD62L and CD69,as well as the absolute number,in CD4+T cell were also analyzed.Results: Compared with WT mice,pathological damage in lung tissues was significantly alleviated in miR-7KD mice.Real-time PCR analysis showed that the relative expression of IL-6 was obviously reduced (P<0.01),conversely,relative expression of IL-4 and TGF-βwere obviously increased (P<0.05).Furthermore,the total cell number in BAL also reduced significantly (P<0.05).Importantly,FACS analysis showed that the proportion and the absolute number of F4/80+Mφcells obviously reduced (P<0.05);however,the proportion of γδT cells increased (P<0.05).Moreover,the proportion and the absolute number of CD4+T cells and CD8+T cells were significantly reduced (P<0.05). Finally, the proportion and the absolute number of CD62L+in CD4+T cells were upregulated vigorously,contrastly,the proportion and the absolute number of CD69+in CD4+T cells were notably up-regulated (P<0.05).Conclusion:miR-7 defeciency could significantly ameliorate the pathology of murine ALI,suggesting that it may play an important regulatory role in the development of ALI.

Objective:To detect the change of proportions of αβT lymphocytes in mesenteric lymph nodes of miR-7 knockdown (miR-7KD)mice,and preliminarily explore its importance.Methods: The volume,weight index and total cells number of mesenteric lymph node in miR-7KD mice were measured.The pathologic morphology change of mesenteric lymph nodes was observed by HE stai -ning.And the changes on proportion of αβT lymphocytes in mesenteric lymph nodes of miR-7KD mice were analyzed by Flow cytometry.Results:Compared with those of WT (wild type) mice,the volume,weight index,and the total cells number of mesenteric lymph node were significantly increased ( P<0.05 ).Moreover ,the pathologic morphology was significantly changed.The proportion and numbers of T lymphocyte were significantly increased;however,the proportion of B lymphocyte were significantly decreased (P<0.05). Notably,the proportion and number of CD4+T cells and CD8+T cells were significantly increased (P<0.05).Meanwhile,CD62L+T cell proportion were vigorously reduced and CD 69+T cell and IFN-γ+T cell proportion were notably up-regulated,which belong to CD4+and CD8+T cell(P<0.05).Conclusion:There was significant influence in the proportion of αβT lymphocytes in mesenteric lymph nodes of miR-7KD mice,suggesting that miR-7 might play an important role in the composition and function of lymphocytes in mesenteric lymph nodes.

Objective:To study the relationship between neutrophil to lymphocyte ratio (N/L) and traditional cardio‐vascular risk factors among community 35~64‐year‐old residents .Methods :A total of 1884 residents (548 males and 1336 females) from urban Shenyang city received baseline condition questionnaire on cardiovascular diseases and re‐lated diseases from Apr 2011 to Feb 2012. According to presence of cardiovascular risk factors or not ,subjects were divided into healthy control group (n=675) and risk factor group (n=1209);according to number of risk factors , risk factor group was further divided into one risk factor group (n=491) ,two risk factors group (n=263) and ≥3 risk factors group (n=455) .Morning blood sample and urine sample were retained to measure blood and urine rou‐tine ,blood glucose and blood lipid profile etc in all subjects .N/L was compared and analyzed among all groups .Re‐sults:Among patients with only one of following risk factors [hypertension ,diabetes mellitus (DM) ,dyslipidemia and obesity] ,N/L levels of patients with hypertension or DM were significantly higher than that of healthy control group [1.55(1.15 ,1.95) ,1.60(1.21 ,2.07) vs .1.45(1.09 ,1.91)] , P<0.05 both ,and there were no significant difference between any other one risk factor group and healthy control group , P>0.05 all .Among risk factor sub‐groups ,N/L level of ≥3 risk factors group was significantly higher than that of two risk factors group [1.57(1.16 , 2.04) vs .1.41(1.07 ,1.89) ,P<0.05] ,and there was no significant difference between any other two groups (P>0.05) .Conclusion:N/L significantly related to hypertension or DM ,and N/L level of ≥3 risk factors group was sig‐nificantly higher than that of two risk factors group ,N/L is helpful to assess risk of cardiovascular diseases .