Osthole attenuates diabetes-induced renal injury by regulating NF-κB and p38/MAPK pathway mediated inflammatory responses / 中国临床药理学与治疗学

To investigate the therapeutic effects of oral osthole on streptozotocin (STZ)-induced type 1 diabetes mellitus(T1DM) mice and explore its internal mechanism. METHODS: The diabetes model induced by STZ was established. Mice were randomly divided into control group, STZ model group, STZ+osthole group (20 mg/kg). Body weight, blood glucose, urine protein, blood urea nitrogen and creatinine were observed to detect renal function. The degree of renal tissue damage was detected by H&E staining and PAS staining, and the degree of renal fibrosis was detected by Sirius Red staining. CD68 and F4/80 immunofluorescence staining was used to observe the infiltration of macrophages in kidney tissue. The mRNA expressive levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) in renal tissue were detected by RT-qPCR. The protein expressive levels of phospho-NF-κB p65, NF-κB p65, IκBα, phospho-IκBα, phospho-p38 and p38 were detected by Western blot in renal tissue. RESULTS: Compared with the STZ model group, the levels of urinary protein, blood urea nitrogen, creatinine were significantly decreased after osthole treatment (P<0.05 or P<0.01). The renal structure disorder, mesangial matrix area, collagen fiber accumulation, and macrophage infiltration were significantly improved (P<0.05 or P<0.01). The expression of mRNA of pro-inflammatory cytokines TNF-α and IL-6 were significantly decreased (P<0.05 or P<0.001). The expression of phospho-NF-κB p65, phospho-IκBα and phospho-p38 were significantly down-regulated (P<0.05 or P<0.01), while the protein expression level of NF-κB p65, IκBα was up-regulated (P<0.05). CONCLUSION: Osthole has a protective effect on kidney injury caused by diabetes and inhibits NF-κB and p38/MAPK signaling pathway.

Inhibition of the replication of H9N2 influenza virus in vivo by short-term repeated oral administration of chicken interferon α / 生物工程学报

To evaluate the optimal administration frequency for interferon-α (IFN-α) and the effect of its combined use with inactive virus on chicken flocks, the prokaryotic expression plasmid pET-22b-ChIFN-α was constructed and transferred into Escherichia coli BL21(DE3) host bacteria to induce the expression of chicken IFN-α and to harvest recombinant proteins inclusion bodies. The expression of recombinant chicken IFN-α was confirmed by SDS-PAGE, and the results demonstrated that the chicken IFN-α (20 kDa) was highly expressed using the prokaryotic expression vector with a concentration of 0.2 mg/mL in the medium. Chicken IFN-α was diluted to 2.5×10⁴ U/fowls and administered to immunized specific-pathogen-free chickens orally in combination with inactivated H9N2 subtype influenza virus. Chicken that received chicken IFN-α were safe after three repeated immunizations (96 h). In addition, chicken IFN-α could induce higher levels of antiviral-related inducible genes in peripheral blood, spleen, and thymus of chicken flocks. The results of a challenge assay revealed that the lowest detoxification rates of chicken IFN-α ranged from three to five days, suggesting a higher capacity to resist H9N2 subtype avian influenza virus. The present study obtained the optimal immune frequency and immunization period for chicken IFN-α to provide theoretical support for the optimal clinical application of IFN-α.

Expression level of lncRNA-ENST00000460164 in luminal A breast cancer and its effect on cell cycle / 基础医学与临床

Objective To find the expression level and the role of ENST00000460164 in luminal A breast cancer.Methods The expression level of ENST00000460164 in breast cancer tissues was detected by RT-qPCR.pll3.7-ENST00000460164-shRNA and empty vector,pll3.7,were transfected into MCF-7 cells.Cell cycle was measured by flow cytometry.Western blot was used to detect the expression of P16INK4A and cyclinD1.Results ENST00000460164 was highly expressed in luminal A breast cancer tissues as compared to the adjacent non-cancer tissues.The knockdown of ENST00000460164 resulted in the G1 cell-cycle arrested,cyclin D1 downregulated and P16INK4A upregulated in MCF-7 cells.Conclusions ENST00000460164 is overexpressed in luminal A breast cancer.ENST00000460164 may control G1/S transition by regulating P16INK4A or cyclin D1 expression.

Ways for establishing a rat model of spinal cord contusion：a network meta-analysis / 中国组织工程研究

BACKGROUND: Animal experimental study on spinal cord injury used injury mode and similarity of clinical spinal cord injury as an important reference index of selecting modeling approach. OBJECTIVE: To compare the difference among the use of precision impactor, homemade Al en’s impactor, spinal cord compression method and clamping method in rat models of spinal cord injury, and to provide a new basis for the selection of the modeling method of a rat model. METHODS: PubMed, CNKI, Wanfang and VIP databases were retrieved with computer from Building to June 20, 2015. Eligible literatures were included and analyzed by ADDIS software. RESULTS AND CONCLUSION: A total of 26 studies met inclusion criteria, containing 599 rats. After analysis of the inclusion studies, the model could be effectively made in each modeling method. The modeling method with effects from best to poor is as fol ows in order: precision impactor, clamping method, homemade Al en’s impactor and spinal cord compression method. According to the lowest mortality, there were precision impactor, homemade Al en’s impactor and clamping method. From the point of view of function and mortality, the use of precision impactor is the best. The use of homemade Al en’s impactor is the most economical. The clamp method could achieve a balance between them.

Effect of apolipoprotein E deficiency on astrocyte destruction / 实用医学杂志

Objective To explore the effect of apolipoprotein E deficiency on astrocyte destruction in the ex vivo spinal cord slice model. Methods Vibratome-cut transverse spinal cord slices from C57BL/6 postnatal day 7 ApoE -/-mice and wild-type mice were cultured on transwell porous supports. After 7 days in the culture , spinal cord slices were exposed to LPS for 5 days. The expression of AQP4, GFAP and Iba1 was detected by immunofluorescence. The concentration of TNF-α and NO were detected by ELISA and method of nitrate reductase, respectively. Results Marked loss of AQP4 and GFAP staining were shown in LPS-affected groups, not in the control group, and the lesion score was higher in ApoE-/-group than WT-group (P<0.05). In addition, the expression of Iba1 and concentration of TNF-α, NO in the LPS-affected groups were more than that of the control group (P < 0.05), and they were higher in the ApoE-/-LPS group than the WT-LPS group (P < 0.05). Conclusions ApoE deficiency exacerbates astrocyte injury, likely through promoting the release of pro-inflammatory mediators from microglia.

The relationship between the neuromyelitis optica astrocyte damage and the patients with functional defect / 实用医学杂志

Objective To explore the relationship between the damage of neuromyelitis optica (NMO) astrocytes (AS) and the onset of NMO, and investigate the relevance of AS damage with the severity of the patients with functional defect. Methods The levels of aquaprin4-antibody (AQP4 -Ab), glial fibrillary acidic protein (GFAP), apolipoprotein(ApoE),interleukins-6(IL-6), interleukins-10(IL-10), tumor necrosis factor-α(TNF-α) in cerebrospinal fluid (CSF ) and serum of 30 acute NMO patients were tested by means of ELISA. The results were later compared with control group. And analysis of the relevance of the various index of the levels in CSF with the CSF AQP4-Ab level, the acute phase expanded disability status scale(EDSS) score of the NMO group were made. Results (1)The NMO group in CSF and serum AQP4-Ab, GFAP, IL-6 levels were higher than the control group (P < 0.05), and ApoE, IL-10 levels were lower than the control group (P < 0.05). (2)The CSF GFAP, ApoE, IL-6 in NMO group is higher than the serum (P < 0.05), and CSF AQP4-Ab, IL-10 levels were lower than the serum ( P < 0 . 05 ) . ( 3 ) The CSF GFAP , IL-6 levels and the CSF AQP4-Ab level were positively correlated (r=0.749, r=0.526, P<0.05), and the CSF ApoE, IL-10 levels were negatively correlated with CSF AQP4-Ab level(r = -0.571, r = -0.676, P < 0.05). (4)The CSF AQP4-Ab, GFAP, IL-6 levels and the acute phase EDSS score were positively correlated (P < 0.05), the CSF ApoE, IL-10 levels were negatively correlated with the acute phase EDSS score (P < 0.05). Conclusion The AS damage exists in the NMO and the damage severity may correlate with patient function defect. AQP4-Ab, GFAP, IL-6 may play important roles in the onset of NMO and the disease aggravating. The decrease of the ApoE and IL-10 may exacerbate NMO damage.

The systematic evaluation of establishing spinal cord transection model in rats / 中国组织工程研究

BACKGROUND:Establishing a highly successful, safe, reliable standard spinal cord transection model is the precondition of studying spinal cord injury repair. OBJECTIVE:To evaluate the value of preparing spinal cord transection model in rats and the effects of laminectomy on spinal cord. METHODS: We searched the randomized controled trials involving rat models of spinal cord transection in the databases of PubMed, CNKI, VIP and WanFang. RESULTS AND CONCLUSION:11 randomized controled trials (RCTs) met the inclusion criteria (two in English, 9 in Chinese), and a total of 394 rats were included in the study. There were significant differences in the lower limb motor function scores (BBB scores) within 1-6 weeks after injury (WMD=-12.86, 95%CI-16.10 to-9.62,P < 0.01) and electrophysiological indices within 4 weeks after injury (WMD=15.36, 95%CI 11.36 to 19.36,P < 0.01) between spinal cord hemisection group and laminectomy group. The BBB scores after 6 weeks were not significantly different between these two groups (WMD=-10.28; 95%CI-24.20 to 3.64;P=0.15). There were significant differences in the lower limb motor function scores (BBB scores) within 1-6 weeks after injury (WMD=