OBJECTIVE:To study the pharmacodynamic effects of volatile oil from Homalomena occulta on adjuvant-induced arthritis(AA)model rats and its mechanism. METHODS:60 rats were randomly divided into normal control group(0.5%polysor-bate 80),model control group (0.5% polysorbate 80),positive control group (Tripterygium glycosides tablets,10 mg/kg), high-dose,middle-dose and low-dose(0.08,0.04 and 0.02 ml/kg)groups of volatile oil from H. occulta. Except for normal control group,other groups were given complete Freund's adjuvant subcutaneously via right rear foot plantar to induce AA model,and giv-en relevant medicine intragastrically for 25 days,once a day,since modeling. The articular swelling degree,immune organ (thy-mus gland and spleen) index,pathological change,the contents of IL-1β and TNF-α in serum were detected. RESULTS:Com-pared with normal control group,the primary and secondary articular swelling degree and thymus gland index of rats and the serum contents of IL-1β and TNF-α increased in model control group,while spleen index decreased(P<0.05 or P<0.01);obvious tissue swelling,large amount of neutrophile granulocyte,leukomonocyte and macrophage infiltrating joint surrounding tissue,the prolifer-ation of synovial cells and obvious osteoarthritic lesion were observed in podarthrum. Compared with model control group,the pri-mary and secondary articular swelling degree and the serum content of IL-1β decreased in the volatile oil from H. occulta groups;thymus gland index increased in middle-dose and low-dose groups of the volatile oil from H. occulta;the content of TNF-α de-creased in high-dose group of the volatile oil from H. occulta(P<0.05 or P<0.01). The inflammatory cell infiltration of joint sur-rounding tissue relieved in treatment groups,synovial cells proliferation was not obvious and synovial cells morphology was im-proved. CONCLUSIONS:The volatile oil from H. occulta has the pharmacodynamic effects on AA in rat,and its mechanism might be related to the serum content reduction of IL-1β and TNF-α.

Objective To evaluate the mutagenicity of Schistosoma japonicum bivalent DNA vaccine.Methods The mutagenicity of Schistosoma japonicum bivalent DNA vaccine was studied by Ames test,micronucleus test and chromosome aberration test.Results On the conditions of actived and unactived,Schistosoma japonicum bivalent DNA vaccine did not lead obvious increase of colony units,and the result of the Ames test was negative.Compared with the control group,there were no significant differences in the treated groups on the frequency of micronucleus in PCE or chromosome aberration.Conclusion There is no mutagenicity of Schistosoma japonicum bivalent DNA vaccine.

Objective To investigate the in vitro expressions of chamotatic factor CXCL12 [also known as stromal cell-derived factor 1 (SDF-1)],the receptor of the factor,Cys-X-Cys receptor 4 (CXCR4) and vascular endothelial growth factor C (VEGF-C) in human laryngeal carcinoma cell line Hep-2 and their roles in the homing of laryngeal carcinoma. Methods The expressions of CXCR4 mRNA and protein in Hep-2 cells was detected by RT-PCR and Western blotting,the expression of VEGF-C also was evaluated before and after incubated with SDF-1 (1,10,100 ng/ml) or the antagonist of CXCR4 AMD3100. Methythiazolyltetrazolium (MTT) assay was used to analyze the effect of different concentrations of SDF-1 on the proliferation of Hep-2 cells. Transwell invasion chamber and matrigel were used to evaluate the effect of various concentrations of SDF-1 and AMD3100 on the migration and invasion of Hep-2 cells. Results 1)The CXCR4 and VEGF-C were both overexpressed at mRNA and protein level in Hep-2 cells,and the expression of VEGF-C in Hep-2 cells was up-regulated with a concentration-dependent model,which was inhibited by CXCR4 antagonist (P

Objective To investigate the effects of sevoflurane post-conditioning on oxidative stress and inflammatory reaction during rat cerebral ischemia-reperfusion, and to explore its cerebral protective mechanism.Methods Thirty-six health male Sprague-Dawley rats (aged 12-14 weeks, weighing 220-260 g) were randomly divided into 3 groups (n=12 each): sham control group (group Sham), cerebral ischemia-reperfusion group (group IR), sevoflurane post-conditioning group (group SPC).Cerebral ischemia-reperfusion model was established, ischemia for 30 min followed by reperfusion 24 h.Rat middle cerebral artery was not occluded in group Sham.Cerebral ischemia-reperfusion model was established in group IR.Group SPC was subjected to 2.6% sevoflurane for 15 min in the beginning of reperfusion.At the end of reperfusion, rats were cut off the head to take out the brain tissue.The expression level of Iba-1 and HO-1 proteins was measured by western blot.The levels of reactive oxygen species (ROS), malondialdehyde (MDA), TNF-α, IL-1β and the activity of superoxide dismutase (SOD) were evaluated.Results Compared with group Sham, the expression of cerebral cortex Iba-1 protein was higher than that in groups IR and SPC (P＜0.05), the expression of Iba-1 protein in group SPC was lower than that in group IR (P＜0.05).Compared with group Sham, the contents of ROS, MDA, TNF-α and IL-1β were increased in groups IR and SPC (P＜0.05), but the activity of SOD and expression of HO-1 protein were decreased (P＜0.05).And the contents of ROS, MDA, TNF-α and IL-1β in group SPC were less than those in group IR, the activity of SOD and expression of HO-1 protein in group SPC were higher than those in group IR.Conclusion Sevoflurane post-conditioning can mitigate the microglia activation, reduce cerebral oxidative stress and inflammation, thus protect rat cerebral against ischemia reperfusion injury.

Objective To study the effects of taking clopidogrel on relevant indicators of platelet aggregation function in 138 cases acute coronary syndrome (ACS) .Methods The platelet function analyzer and flow cytometry were adopted to detect the ADP‐induced platelet aggregation rate ,P selectin and activated GP Ⅱ b/ Ⅲ before medication and on 7 d after taking clopidogrel . Results The platelet aggregation rate after taking clopidogrel for 7 continuous d was decreased significantly (P<0 .01);the P se‐lectin level and activated GP Ⅱ b/ Ⅲ a expressed on platelet surface were significantly reduced (P<0 .01) as well .Conclusion Taking clopidogrel could reduce the platelet aggregation significantly in the patients with ACS and has the effect for inhibiting the platelet aggregation .

Objective To observe the curative effect of low molecular heparin for treating secondary high coagulation state in the patients with nephrotic syndrome(NS) .Methods Total 87 cases of NS in our hospital were divided into the conventional treat‐ment group (n=42) and the low molecular heparin treatment group (n=45) .The routine treatment group was given the prednisone treatment and the low molecular heparin treatment group was treated by low molecular heparin combined with prednisone .The re‐lated indicators of blood coagulation before and after treatment were detected and the clinical curative effects in two groups were an‐alyzed .Results The coagulation related indicators in the conventional treatment group had no statistically significant difference be‐tween before and after treatment (P>0 .05) ,the prothrombin time(PT) and activated partial thrombin time(APTT) after treat‐ment in the low molecular heparin treatment group were significantly extended compared with before treatment ,while the concen‐trations of D‐dimer and fibrinogen were significantly decreased and the concentration of antithrombin Ⅲ was markedly increased compared with before treatment ,showing statistically significant differences between the two groups (P<0 .05);the patients of the low molecular heparin group patients had no bleeding after treatment .Conclusion Low molecular heparin combined with predni‐sone can reduce the secondary high condensation state in NS without bleeding and has a significantly clinical effect .

Objective:To prepare a bone substitute using microsphere scaffold containing adiponectin(APN)and to investigate the release behavior of the scaffold in vitro.Methods:Chitosan microsphere was developed by an emulsion-ionic cross-linking method. Poly (L-lactic-co-glycolic)acid (PLGA)and β-tricalcium phosphate (β-TCP)were used to prepare microsphere scaffold containing APN.The morphology,particle size,drug loading,incorporation efficiency and release behavior of the microsphere were examined. Results:The APN containing microsphere showed good spherical geometry,suitable size and microporosity under scanning electron microscope.The average diameter of the milipore was 20 -200 μm;the drug loading and incorporation efficiency were 1 .3% and 70.3% respectively.The controled-release process continued for 91 days.The extract solution from the APN microsphere-scaffold promoted MC3T3 cell proliferation without cytotoxicity.Conclusion:The APN microsphere-scaffold has sustained release function and may promote osteoblast proliferation.

Objective To investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on voltage-gated calcium channels on the cell membrane of pheochromocytoma cell 12 (PC 12 cells) induced by ischemic/hypoxia and its mechanisms.Methods PC12 cells at logarithmic phase were collected,which were challenged with hypoxia for 6 hours and reoxygenation for 12 hours to stimulate the nerve cells suffered ischemia/reperfusion pathological process in vitro.PC12 cells were divided into non-infection group,infected by lentivirus containing green fluorescent protein (GFP) without HSP70 gene lentivirus control group (GFP lentivirus control group),and infected by lentivirus containing HSP70 and GFP gene recombined lentiviral infection group (HSP70 recombined lentiviral infection group).Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot were used to determine mRNA and protein expressions of L-type voltage-gated Ca2+ channel subunits cav1.2 and cav1.3,receptor gated channel subunits NR1 and NR2,and Na+/Ca2+ exchange (NCX) in PC12 cells.Results After being challenged with hypoxia/reoxygenation,the mRNA and protein expressions of cav1.2,NR1 and NR2 in the PC12 cells were significantly lower in HSP70 recombined lentiviral infection group than those of GFP lentivirus control group and non-infection group [cav1.2 mRNA (2-△△Ct):3.13 ± 0.46 vs.5.12 ± 0.52,5.13 ± 0.66;NR1 mRNA (2-△△△Ct):1.61 ± 0.44 vs.3.23 ±0.82,3.31 ±0.78;NR2 mRNA (2-△△Ct):2.09±0.41 vs.3.91 ±0.64,3.88±0.62;cav1.2 protein (gray value):2.82±0.39 vs.3.98±0.23,3.96±0.24;NR1 protein (gray value):1.84±0.35 vs.2.79±0.21,2.86±0.23;NR2 protein (gray value):0.87±0.24 vs.1.57±0.31,1.33±0.44;all P ＜ 0.01].But there were no statistical differences in the mRNA and protein expressions of cav1.2,NR1 and NR2 between GFP lentivirus control group and non-infection group (all P ＞ 0.01).There were no statistical differences in the mRNA and protein expressions of cav1.3 and NCX among non-infection group,GFP lentivirus control group and HSP70 recombined lentiviral infection group [cav1.3 mRNA (2-△△Ct):4.82 ± 0.32,4.72 ± 0.36,4.82 ± 0.29;NCX mRNA (2-△△Ct):3.49 ± 0.78,3.47 ± 0.71,3.56 ± 0.65;cav 1.3 protein (gray value):2.63±0.40,2.64±0.39,2.68±0.39;NCX protein (gray value):3.27±0.48,3.34±0.48,3.31 ±0.42;all P ＞ 0.01].Conclusion Exogenous HSP70 affects the expression of L-type voltage-gated Ca2+ channel subunit cav1.2 and receptor gated channel subunits NR1 and NR2,which may protect PC12 cells from the injury caused by ischemic/hypoxia.

Objective To investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on calcium homeostasis and calcium channels of PC12 cells induced by ischemic and hypoxia and its mechanisms. Methods PC12 cells at logarithmic phase were collected, and they were divided into recombined lentiviral infection group [infected by lentivirus containing HSP70 and green fluorescent protein (GFP) fluorescin gene], lentivirus control group (infected by lentivirus containing GFP without HSP70 gene) and non-infection group. PC12 cells were subjected ischemia/hypoxia for 4, 8, 12, 24 hours, and the cell activity was determined by methylthiazolyl tetrazolium (MTT) assay test inorder to determine the best time for ischemia/hypoxia. The mRNA expressions of HSP70, muscle/endoplasmic reticulum Ca2+-ATP isoforms (SERCA2a, SERCA2b), ryanodine receptor 2 (RyR2), and inositol 1,4,5-triphosphate receptor 1 (IP3R1) were determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and the protein expressions of HSP70, SERCA, and IP3R were determined by Western Blot at 8 hours after ischemic/hypoxia. Flow cytometry was used to determine the levels of intracellular reactive oxygen (ROS) and intracellular Ca2+ ([Ca2+]i). Results With the prolongation of time of ischemia/hypoxia, the cell viability in all groups showed an increase followed by a weakening, and peaked at 8 hours. The cell viability at 8 hours in lentiviral infection group was significantly higher than that of the non-infection group and lentivirus control group [A value (×10-2): 20.3±2.2 vs. 14.1±2.1, 15.0±1.6, both P < 0.01], the mRNA and protein expressions of HSP70 and SERCA in lentiviral infection group were significantly increased [HSP70 mRNA (2-ΔΔCt ): 0.785±0.018 vs. 0.428±0.019, 0.423±0.023; HSP70 protein (gray value): 2.72±0.20 vs. 1.56±0.36, 1.63±0.41; SERCA2a mRNA (2-ΔΔCt ): 0.971±0.037 vs. 0.367±0.014, 0.347±0.012; SERCA2b mRNA (2-ΔΔCt ): 8.869±0.162 vs. 3.015±0.091, 2.941±0.091; SERCA protein (gray value): 2.84±0.18 vs. 1.48±0.26, 1.52±0.29], and IP3R2 mRNA and protein expressions were significantly declined [IP3R2 mRNA (2-ΔΔCt ): 0.183±0.020 vs. 0.439±0.020, 0.433±0.040; IP3R2 protein (gray value): 1.15±0.12 vs. 1.91±0.20, 1.83±0.19], with statistically significant differences (all P < 0.01); no significant difference in RyR mRNA was found [2-ΔΔCt (×10-3): 1.97±0.63 vs. 2.02±0.22, 2.01±0.09, both P > 0.05]; the relative fluorescence intensity of ROS and [Ca2+]i in lentiviral infection group was significantly reduced (ROS: 30.54±1.23 vs. 58.03±1.97, 57.72±2.35; [Ca2+]i: 34.50±2.05 vs. 48.20±3.02, 46.80±2.75, all P < 0.01]. Conclusion Exogenous HSP70 can maintain calcium homeostasis in the endoplasmic reticulum of PC12 cells, affect the Ca2+ channel protein regulated by calcium channel IP3R and calcium pump SERCA, which may cause hypoxia/ischemia intracellular injury.

·AIM:To present a case of late onset bilateral keratectasis after laser in situ keratomileusis (LASIK) for myopia with rigid gas-permeable contact lenses with a brief review of literature on this subject.·METHODS:A 27-year-old woman underwent bilateral uneventful LASIK for moderate myopia. Preoperative cycloplegic refractions were -5.50/-0.50×50° right eye (OD) and - 4.50/-1.00×15° left eye (OS).Corneal pachymetry was 526μm OD and 541μm OS, Preoperative corneal topography was normal and did not reveal any keratoconus or forme fruste keratoconus.Following the creation of flaps with 160μm plates,ablations of 102μm OD and 86μm OS were performed,estimated to leave residual stromal beds of 264μm OD and 295μm OS.·RESULTS:Twenty-nine months postoperatively,the patient developed bilateral inferior keratectasia of -12.50/-4.00×160° OD and -6.00/- 4.25×125° OS.Visual acuity was reduced in both eyes;the central cornea had steepened; and pachymetry showed central corneal thinning.Keratectasia was diagnosed,and rigid contact lenses were fitted.Three years later,the patient achieved satisfactory visual acuity and all-day lens wear with minimal complications.·CONCLUSION:Late keratectasia may follow LASIK for low to moderate myopia despite a thorough preoperative work-up.Rigid contact lenses can offer a safe,reversible option for improving visual acuity in such patients by delaying or avoiding the need for intracorneal ring segments implanting or penetrating keratoplasty.