OBJECTIVES: To investigate the effects of formulated granules of Tianma Gouteng Yin (TGY) on motor deficits in a mouse model of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced subacute Parkinson's disease (PD) and explore the possible molecular mechanisms. METHODS: Ninety C57BL/6 mice were randomized equally into 6 groups, including a control group, a PD model group, a NEC-1 (6.5 mg/kg) treatment group, two TGY treatment groups at 5 and 2.5 g/kg, and a Madopar (76 mg/kg) treatment (positive control) group. Mouse models of PD were established by intraperitoneal injection of MPTP (30 mg/kg) for 5 consecutive days with the corresponding treatments for 15 days. The mice were randomly selected for motor function tests. Western blotting was used to detect the changes in expressions of TH, α-syn, RIPK1, RIPK3 and MLKL in the striatum of the mice. Network pharmacology analysis and molecular docking studies were performed to explore TGY-mediated regulation of the necroptosis pathway for PD treatment. RESULTS: Compared with those in the control group, the PD model mice exhibited obvious motor deficits with significantly increased α-syn protein expression and lowered TH protein expression in the striatum. Treatment with NEC-1 obviously improved motor deficits, inhibited the necroptosis pathway, and alleviated the changes in TH and α‑syn proteins in PD mice. Network pharmacology and molecular docking analyses suggested that the therapeutic effect of TGY in PD was associated with the modulation of RIPK1, a key protein in the necroptosis pathway. In PD mouse models, TGY treatment at the two doses significantly improved motor deficits of the mice, increased TH expression, and decreased the expressions of α-syn and necroptosis-related proteins in the striatum. CONCLUSIONS TGY can effectively inhibit the necroptosis pathway, increase TH expression and decrease α-syn expression in the striatum to improve motor deficits in PD mice.
Animals ; Mice, Inbred C57BL ; Mice ; Necroptosis/drug effects* ; Drugs, Chinese Herbal/therapeutic use* ; Parkinson Disease/drug therapy* ; Disease Models, Animal ; Male

Objective To investigate the potential mechanism by which dihydromyricetin(DHM)ameliorates diabetic nephropathy(DN),and to screen and validate its possible key molecular targets.Methods A DN model was established using db/db mice,and 100 mg/(kg·d)DHM was administered via gavage 5 d per week for totally 10 weeks.Renal morphological changes were observed after staining to evaluate the effects of DHM.GSE161885 and GSE270526 datasets were obtained from the Gene Expression Omnibus(GEO)database and analyzed in combination with the GeneCards database to screen for DN-related differentially expressed genes(DEGs).Protein-protein interaction(PPI)network and molecular docking were employed to predict potential DHM targets.Western blotting and immunofluorescence staining were performed to detect the effects of DHM on pyroptosis-related pathways in the renal tissues of db/db mice and in high glucose(HG)-induced human renal tubular epithelial cells(HK-2).The specific NLR family pyrin domain containing protein 3(NLRP3)inhibitor MCC950 was also used to validate the predicted mechanism.Results In vivo experiments showed that DHM significantly ameliorated renal pathological damage in db/db mice,alleviated glomerular hypertrophy and mesangial expansion,and markedly reduced Paller scores(P<0.001).Immunofluorescence staining revealed significantly weakened fluorescence signals for α-smooth muscle actin(α-SMA),fibronectin,and collagen Ⅰ in renal tissues.Western blot results showed that the expression levels of collagen Ⅰ,collagen Ⅲ,α-SMA,and transforming growth factor beta 1(TGF-β1)were significantly decreased(P<0.05).A total of 16 DN-related DEGs were identified.Enrichment analysis revealed that these genes were primarily enriched in pathways such as viral protein interactions,cytokine-cytokine receptor interaction,and the AGE-RAGE signaling pathway in diabetic complications,and were primarily involved in gene functions such as the positive regulation of lymphocyte-mediated immunity,positive regulation of adaptive immune response,and chemokine activity.Molecular docking confirmed NLRP3 as a potential target of DHM.In vivo validation showed that DHM significantly down-regulated gasdermin-D(GSDMD)fluorescence signals and inhibited the expression of pyroptosis-related proteins including NLRP3,Caspase 1,Cleaved-Caspase 1,interleukin 18(IL-18),and GSDMD(P<0.05).In vitro studies further confirmed that both DHM and the specific NLRP3 inhibitor MCC950 alleviate high glucose-induced fibrosis and pyroptosis in HIC-2 cells.Conclusion DHM can ameliorate the progression of DN,and its mechanism is related to inhibiting NLRP3 inflammasome-mediated pyroptosis,thereby alleviating renal inflammation and fibrosis.

As a potential vectored vaccine,Newcastle disease virus(NDV)has been subject to various studies for vaccine development,while relatively little research has outlined the immunomodulatory effect of the virus in antigen presentation.To elucidate the key inhibitory factor in regulating the interaction of infected dendritic cells(DCs)and T cells,DCs were pretreated with the NDV vaccine strain LaSota as an inhibitor and stimulated with lipopolysaccharide(LPS)for further detection by enzyme-linked immunosorbent assay(ELISA),flow cytometry,immunoblotting,and quantitative real-time polymerase chain reaction(qRT-PCR).The results revealed that NDV infection resulted in the inhibition of interleukin(IL)-12p40 in DCs through a p38 mitogen-activated protein kinase(MAPK)-dependent manner,thus inhibiting the synthesis of IL-12p70,leading to the reduction in T cell proliferation and the secretion of interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),and IL-6 induced by DCs.Consequently,downregulated cytokines accelerated the infection and viral transmission from DCs to T cells.Furthermore,several other strains of NDV also exhibited inhibitory activity.The current study reveals that NDV can modulate the intensity of the innate?adaptive immune cell crosstalk critically toward viral invasion improvement,highlighting a novel mechanism of virus-induced immunosuppression and providing new perspectives on the improvement of NDV-vectored vaccine.

The initiation of tumor is a complex process with multi-factor participation, particularly the activation of oncogenes and/or inactivation of tumor suppressor genes. Long non-coding RNAs (lncRNAs) play important roles in tumorigenesis. Additionally, as a metabolic process in cells, autophagy also contributes greatly to differentiation, metastasis and chemoresistance of tumor cells, and has become a central topic in recent years. The understanding of connection between lncRNAs and autophagy as well as their mechanisms underlying tumorigenesis, can provide new ideas for the diagnosis and treatment of tumors.

Objective To explore the application effect of PDCA cycles on the normal limb position of stroke patients with hemiplegia. Methods The stroke patients with hemiplegia (128 cases) were selected as study subjects. Patients(62 cases)during January to September 2014 were set as the control group,and received routine nursing care. Patients(66 cases)during October 2014 to July 2015 were set as the experimental group,and used PDCA cycles management on the normal limb position additionally. The application effect of PDCA cycles on the normal limb position was evaluated through comparing two groups with qualification rates of normal limb position and incidence of complications. Results The qualification rates of normal limb position in the control group was 38.71%(24/62), which was higher than that of the control group, which was 75.76% (50/66) (χ2=16.504, P<0.01). The complications occurred in the control group were strephenopodia (11 cases), foot drop (16 cases), dislocation of shoulder (9 cases), omodynia (27 cases) and myospasm (34 cases), and they were 3 cases, 7 cases, 2 cases, 15 cases and 18 cases in the experimental group respectively. The incidence of complications was lower than those of the control group (χ2=4.001-8.961, P < 0.05), and the difference was statistically significant between two groups. Conclusions PDCA cycles management could improve the qualification rates of normal limb position and reduce the incidence of complications, which was beneficial to the recovery of limb function.

Objective To establish a method of SYBR Green Ⅰ FQ-PCR for detecting the expression of miR-202 in peripheral blood mononuclear cells ( PBMC ) and analyze the expression of miR-202 and its clinical significance in MM.Methods Reverse transcription was performed with specific stemloop primer for miR-202,and then FQ-PCR was used to detected the expression of miR-202 in 21 MM patients and 20 healthy people.Data was presented as mean ± standard deviation ( (x) ± s ).Non-parametric Mann-Whitney test was used to analyze the difference between MM group and control group.In addition,1∶ 125 dilution of one test sample was detected by repeated 5 times,and the same sample was tested one time a day for 3 days,repeated 5 each time.The assessment of repeatability of measurements was done by calculating standard deviation and variation coefficient from threshold cycle (Ct).Results FQ-PCR detection of miR-202 in PBMC was amplified by the standard S-curve,with a single melting curve peak and no complex peak,which showed good specificity.The assay showed good reproducibility (intra-assay coefficient 1.2％ and inter-assay coefficient 3.2％ ) and high sensitivity ( 12.8 pmol/μ1).The expression of miR-202 in MM was 0.014 ( 0.007 - 0.221 ) and 1.844 ( 0.162 - 3.966 ) in normal controls.The expression level of miR-202 was significantly higher in MM than in normal controls (U =48.000,P ＜0.01 ).Conclusions FQ-PCR provide us a rapid,sensitive and specific method for detection of miR-202.The expression level of miR-202 is higher in MM than in normal controls.It is possible to play a role in MM progress and may be a useful marker to evaluate the development and treatment of MM.