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This study aims to investigate the inhibitory effect of Pien Tze Huang(PZH) on enterovirus 71(EV71). To be speci-fic, chemiluminescence method was adopted to evaluate the toxicity of PZH to African green monkey kidney(Vero) cells and human rhabdomyosarcoma(RD) cells, and cytopathic effect(CPE) method to assess the inhibition on EV71-GFP reporter virus and EV71 C4 wild-type virus. The results showed that PZH had low cytotoxicity to Vero cells and RD cells, with the half-maximal cytotoxic concentration(CC_(50)) of about 0.691 3-0.879 2 mg·mL~(-1) for the two. In addition, PZH can effectively inhibit the replication of EV71 within the non-cytotoxic concentration range, and dose-dependently alleviate the cytopathic changes caused by virus infection, with the half-maximal effective concentration(EC_(50)) of 0.009 2-0.106 3 mg·mL~(-1). On the basis of the above results, the green fluorescent protein(GFP), indirect immunofluorescence assay(IFA), and median tissue culture infective dose(TCID_(50)) were employed to assess and verify the anti-EV71-GFP and anti-EV71 C4 activity of PZH. The results demonstrated that PZH can dose-dependently lower the expression of GFP by EV71-GFP and structural protein VP-1 by EV71 C4 and decrease the production of progeny infectious viruses. The EC_(50) of PZH for EV71-GFP and EV71 C4 was about 0.006 0-0.006 2 mg·mL~(-1) and 0.006 6-0.025 6 mg·mL~(-1), respectively. This study suggested that PZH may exert antiviral activity by acting on EV71 and interfering with the expression of VP-1. At the moment, there is still a lack of specific anti-EV71 drugs. This study proposed a new idea for the symptomatic treatment of EV71 infections such as hand-foot-mouth disease and verified an effective drug for the treatment of EV71 infections.
Subject(s)
Animals , Chlorocebus aethiops , Drugs, Chinese Herbal/pharmacology , Enterovirus A, Human/physiology , Hand, Foot and Mouth Disease , Vero CellsABSTRACT
Objective To study the predictive value of serum hyaluronic acid (HA) levels for morphological progression of osteoarthritis (OA).Methods Seventy New Zealand white rabbits were randomly and equally divided into the OA group and the normal control group.OA was induced by injecting 0.5 ml of 2％ papain into the left knee joints and the same volume of sterile saline solution was injected Into right knee joints in control group.The serum HA levels were detected at 1,2,4,6,8,10 and 12 weeks after injection.Then magnetic resonance imaging (MRI) and pathohistological changes were observed.T test was used for the analysis of measurment.Resuts Compared with the control group,degeneration changes were found in the OA group,such as thinner articular cartilage,fibrillation and destroyed cartilage matrix,and inflammation,proliferation and degeneration of the synovial tissue.All these changes were much worse with prolonged observation time.The serum HA levels were increased [12 week:(657±132) ng/ml,vs (166±8) ng/ml],which correlate with structural damage.Conclusion Serum HA levels had a predictive value for further development of OA.
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Objective To observe the influence of different doses of intravenous immunoglobulin (IVIG) on function of T cells in cord blood of premature infants and to explore the immunomodulating mechanism of IVIG.Methods Cord blood mononuclear cells (CBMC) were isolated by density gradient centrifugation method from 15 preterm infants born between June 1,2011 to December 31,2011 at 32 34 gestational weeks.Three groups were formed according to the concentrations of IVIG used for CMBC culturing in vitro (Group A,6 mg/ml; Group B,9 mg/ml; Group C,12 mg/ml).After 72 hours in sterile conditions,the levels of IFN 7,IL 4,IL-10 and TGF β1 in the supernatant were determined by enzyme-linked immunosorbent assay.The expression of CD4+ CD25+ Foxp3+ Treg cells was examined by fluorescent activated cell sorter.Differences among groups were compared by one way analysis of variance.For comparison between two groups,LSD test was used.Results (1) The level of IFN γ secreted by CBMC in Group C was 0.03 ± 0.02,which was much lower than that in Group A(0.18±0.08) and Group B(0.13±0.05) (F=5.72,both P＜0.01).The level of IFN-γ in Group B and Group A did not show statistical difference (P＞0.05).Compared with Group A(0.03±0.01),the level of IL4 was much lower in Group B (0.02±0.01) and Group C(0.01±0.01) (F=20.38,both P＜0.01),while no significant difference was shown between Group B and Group C (P＞0.05).(2) No significant difference was found in the expression of CD4+CD25+Foxp3+Treg cells among different groups (F 0.67,P＞0.05).(3) The level of IL 10 in Group C was 3.02 ± 3.79,which was significantly lower than that in Group A (10.78±5.44) and Group B (6.90±4.64)(F=4.68,P＜0.01 and 0.05).The level of IL-10 inGroup B was still lower than that in Group A (P＜0.05).The levels of TGF-β1 in Group C(8.44± 13.71) and Group B(16.15 ±13.94) were significantly lower than that in Group A(30.23 ± 16.32) (F=5.22,P＜0.01,P＜0.05),but there was no significant difference between Group B and Group C (P＞0.05).Conclusions IVIG might inhibit the function of Th cells in CBMC of preterm infants in a dose dependent manner.IVIG could inhibit the function of natural Treg cells by regulating the secretion of cytokines IL-10 and TGF-β1 in a dose dependent effect.
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Objective To evaluate the value of fat-saturated three-dimensional fast imaging with steady state procession(FS-3D-FISP)sequence in detecting hyaline cartilage defects in osteoarthritis in rabbits.Methods Osteoarthritis was induced in eighteen male rabbits by injecting papain into the left knee joints.The same volume of sterile saline solution was injected into right knee joints as controls.MRI of knee joints was done at 1,2 and 4 weeks after injection using FS-3D-FISP sequence.The the rabbits were killed and pathohistological examinations of the femoral condylars and tibial plateaus were performed.The cartilage changes were observed by MR image according to the Outerbridge method and by pathology using Collins staging.Results There was closely correlation between MR imaging and pathology in grading of cartilage injury in grade I～II and grade III～IV(P＜0.001).Conclusion FS-3D-FISP can evaluate accurately the degree of articular injury.
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0.05).⑥Complications:1 case in the prosthetic disc nucleus replacement group had migration of the device in 1 year after the surgery,but no symptom and did not need revision;1 case complained of bilateral leg pain and 2 cases developed right leg pain in the fusion group,which were treated with oral medications and resolved after the 3-month follow-up.All patients with leg symptoms had surgery at the L5S1 disc space.CONCLUSION:①Quantitative clinical outcomes following prosthetic disc nucleus replacement are at least equivalent to clinical outcomes with traditional lumbar spinal fusion.These results support earlier reports in the literature.②Prosthetic disc nucleus replacement is a safe and effective alternative to fusion for the surgical treatment of symptomatic lumbar disc herniation in properly indicated patients.