The chemokine fractalkine primarily involves in chemotaxis,adherence and inflammation.Recently it has been discovered owning the ability of inhibiting cell death in neurons and glial cells,as well as protecting central nervous system from toxic damage.Fractalkine reduces the toxicity to neurons and glial cells mediated by excessive Fas L,glutamate(Glu) or lipopolysaccharide(LPS).In vivo neutralization of endogenous brain fractalkine signal pathway increases inflammatory cytokines secretion and neuronal cell death induced by LPS.Fractalkine may achieve its neuroprotective property through influencing expression of pro-apoptotic and anti-apoptotic proteins,intracellular Ca2+ level and inflammatory cytokines secretion via protein kinase B(PKB)/Akt and extracellular signal-regulated kinase(ERK)/MAPK pathways.

Objective To evaluate the role of extracellular signal-regulated kinase-cyclic AMP response element binding protein(ERK-CREB) signal pathway in glucocorticoid receptors-mediated chronic morphine tolerance in rats.Methods Male SD rats aged 2 months weighing 280-320 g were used in this study.A catheter was placed in subarachnoid space via foramen magnum according to Yaksh.Thirty-six rats in which intrathecal (IT) catheters were successfully implanted were randomly divided into 6 groups ( n =6 each):control group ( group C),chronic morphine tolerance group (group M),morphine + dexamethasone group (group MD),morphine + RU38486 group (group MR),dexamethasone group (group D),RU38486 group (group R).Normal saline 10 μl,morphine 10 μg,morphine 10 μg + dexamethasone 4 μg,morphine 10 μg + RU38486 2 μg,dexamethasone 4μg,RU38486 2 μg was administered IT twice a day(8:00 and 20:00)for 6 consecutive days in groups C,M,MD,MR,D,R respectively.Tail flick latency (TFL) was measured at 1 d before IT drug administration(baseline)and at 30 min after first IT drug administration during 1,3,5 d and at 1 d after last IT drug administration (T1～4).Maximum analgesic effect (MPE) was calculated.The animals were sacrificed after last TEL measurement.The L3～5 segment of the spinal cord was isolated for determination of the expression of phosphorylated ERK(pERK)and phosphorylated CREB(pCREB) by immunofluorescence staining.Results MPE was significantly T1 lower at T3,4 than at T1 in groups M and MD.Compared with group C,MPE was significantly increased,the expression of pERK and pCREB up-regulated in group M,but no significant change was found in the parameters mentioned above in groups R and D.Compared with group M,MPE was significantly increased,the expression of pERK and pCREB up-regulated in group MR,and MPE was significantly increased,the expression of pERK and pCREB down-regulated in group MD.Conclusion The mechanism by which glucocorticoid receptors-mediated chronic morphine tolerance may be associated with the inhibition of ERK-CREB pathway.

Atrial Flutter ; diagnosis ; etiology ; therapy ; Electrocardiography ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Treatment Outcome

Objective To investigate the analgesic effect of intrathecal(IT)human bone marrow mesenchymal stem cells(hMSC)genetically modified with human proenkephalin gene(PENK)in a rat model of neuropathic pain.Methods Forty male SD rats weighing 160-180 g in which IT catheters were successfully implanted without complication were randomly divided into 4 gorups(n = 10 each): group A normal control;group B neuropathic pain(NP);group C NP + hMSC-pBABE and group D NP + hMSC-PENK.Neuropathic pain was induced with chronic constrictive injury(CCI).Four loose ligatures were placed on the main stem of sciatic nerve with 4-0 chronic catgut.IT normal saline 10 μl,hMSC-pBABE cell suspension 10 μl(2 × 108-3 × 108/μl)and hMSCPENK cell suspension 10 μl(2 × 108-3 × 108/μl)were injected in group B,C and D respectively on the 3rd day after operation.Paw-withdrawal latency(PWL)to noxious thermal stimulation was measured before(baseline)and at 3,5,7,9 and 14 d after operation.The animals were killed on the 14th day after last PWL measurement.RNA was extracted from the spinal cord for determination of proenkephalin mRNA expression.Results PWL was significantly decreased after operation as compared with the baseline values before operation in group B,C and D.PWL was significantly longer at 7,9,14 d after operation in group D than in group B and C but there was no significant difference in PWL after operation between group B and C.PENK mRNA expression was significantly lower in group B and C than in group A,but was significantly higher in group D than in group B and C.There was no significant difference in PENK mRNA expression between group B and C.Conclusion Intratheccal human bone marrow mesenchymal stem cells genetically modified with human proenkephalin gene can relieve neuropathic pain in rats.

Objective To develop an assay for determining bacterial endotoxin(BE T) content in Maodongqing Compound Injection (MCI). Methods The dilution radio o f Maodongqing Compound Injection was optimized and BET content was determined by turbidimetric kinetic method. Results Dilution ratio of Maodongqing Compound In jection at 1 ∶80 did not interfere with the Limulus test. The average recovery of BET was between 50 %and 200 %. Conclusion The method is proved to be accura te,sensitive and suitable for the determination of BET content in Maodongqing C ompound Injection.

OBJECTIVE:To comparison the clinical efficacy and safety of sitagliptin and Repaglinide in the treatment of new-ly diagnosed patients with type 2 diabetes mellitus. METHODS:214 newly diagnosed patients with type 2 diabetes mellitus were randomly divided into study group and control group with 107 cases in each group according to random number table method. Both group received routine diabetes diet,health education and suitable exercise. Control group was treated with Repaglinide tablet 1.0 mg,tid;while study group was treat with Sitagliptin tablet 100 mg,qd. Lab indexes,FBG,PBG,HbA1c and BMI were observed in 2 groups before and after treatment. ADR were compared between 2 groups. RESULTS:There was no statistical significance in lab indexes between 2 groups before and after treatment (P>0.05). After treatment,FBG,PBG,HbA1c were significantly de-creased significantly in 2 groups,with statistical significance (P<0.05),there was no statistical significance between 2 groups (P>0.05). BMI of 2 groups were increased significantly,and the study group was higher than the contrd group,with statistical sighificance (P<0.05). Patients in the study group had no hypoglycemia,while 3 patients in the control group suffered from it, there was statistical significance (P>0.05). CONCLUSIONS:Sitagliptin is similar with repaglinide in therapeutic efficacy;both can effectively reduce blood glucose of elderly patients with newly diagnosed type 2 diabetes mellitus,and the incidence of hypo-glycemia was low,the saterty was good,but sitagliptin has a better control of BMI in patients.

Background and purpose:Recent researches have shown that Secreted Protien Acidic and Rich in Cysteine (SPARC) was closely related to tumor genesis, tumor progression and tumor metastasis. SPARC was highly expressed in malignant melanoma, glioma, meningioma, bladder cancer, lung cancer and prostate cancer, etc. In this study we investigated SPARC expression in hepatocellar carcinoma (HCC) and its signifi cance. Methods:RT-PCR was used to detect SPARC mRNA expression in cancer tissue samples and their adjacent liver tissue samples from 62 patients with hepatocellar carcinoma and 30 normal liver tissue samples, respectively. And the differential protein expression of SPARC between these groups was analyzed by immunohistochemistry (IHC). Results:SPARC mRNA was highly expressed in HCC(14.0?3.6) and in the adjacent liver tissue (6.8?1.8); compared with low expression of 2.7?0.9 in normal liver tissue, there were signifi cant differences among the three groups (p=0.000). SPARC positively stained was found in 54 of 62 patients with HCC and 4 of 30 normal liver tissue, there was significant difference between these two groups (P=0.000). SPARC immunohistochemical score was 21.5?4.8 in the carcinoma group; 11.3?3.6 in paracarcinoma group and 5.7?1.8 in the normal group, there were also significant differences among the three groups (P=0.000). The expression of SPARC protein was significantly upregulated with the progress of Enmondson pathological classification. There was obviously differences between Ⅰvs Ⅱ(P=0.029), and Ⅱ vs Ⅲ Ⅳ(P=0.008). There was more SPARC expression in the patients with metastasis of HCC (26/27, 96.3%) than that without metastasis(23/35, 65.7%)(P=0.004). Conclusion:SPARC mRNA expression and its protein were related to HCC histological differentiation and metastatic lymph node; SPARC is helpful to clinical evaluation of HCC.

Germany ; Hazardous Substances ; analysis ; Workplace

OBJECTIVE:To study the pharmacokinetics of triptolide in Tripterygium glycosides tablet in normal rats and adju-vant arthritis model rats in vivo,and provide reference for clinical rational drug use. METHODS:12 SD rats were randomly divid-ed into normal group and model group,6 in each group. Model group was subcutaneously injected complete Freund's adjuvant 0.1 mL to induce adjuvant arthritis model,normal group was subcutaneously injected the same volume of saline. After 14 d model-ing,2 groups were given Tripterygium glycosides tablet suspension 96 mg/kg intragastrically,the blood sample of eyes 0.4 mL were respectively taken before and 10,30,45,60,90,120,150,180,240,300,420 min after administration. The plasma con-centration of triptolide was determined by HPLC,the pharmacokinetic parameters were calculated by DAS 2.0 software,and the parameters were compared. RESULTS:The pharmacokinetic parameters of triptolide in normal group were cmax of(1.139±0.114)μg/mL,tmax of(2.167±0.606)h,t1/2α of(5.500±3.610)h,AUC0-7 h of(5.052±0.371)μg·h/mL,MRT0-7 h of(3.224±0.119)h, and CL of(11.616±2.986)mL/h;and those in model group were cmax of(0.916±0.103)μg/mL,tmax of(3.083±0.801)h,t1/2αof(5.593±1.795)h,AUC0-7 h of(4.707±0.347)μg·h/mL,MRT0-7 h of(3.429±0.139)h,and CL of(11.246±2.638) mL/h. Compared with normal group,cmax in model group was significantly decreased,tmax and MRT0-7 h were significantly prolonged(P<0.05). CONCLUSIONS:Adjuvant arthritis can affect the pharmacokinetics of triptolide in rats in vivo,and promote its absorption and removal.

Objective To investigate the effects of NB-UVB on the expression of Gadd45α as well as cell apoptosis and cycle of human HaCaT keratinocytes.Methods Cultured HaCaT cells were exposed to various doses (100,200,400 mJ/cm2)of NB-UVB followed by an additional culture of 6,12 and 24 hours,respectively.Reverse transcription PCR and Western blot were performed to detect the mRNA and protein expression of Gadd45α respectively in HaCaT cells,cell counting kit 8 (CCK8)to measure the proliferation of cells,and flow cytometry to determine the cell cycle distribution of HaCaT cells before and after the exposure to NB-UVB.Results Gadd45α was expressed in HaCaT cells.After exposure to NB-UVB of the three doses,the mRNA and protein levels of Gadd45α increased at 6 hours and 12 hours,but declined at 24 hours,and significant changes were observed in HaCaT cells at the three time points after exposure to NB-UVB of the three doses (all P<0.05).The Gadd45α/β-actin mRNA ratio was 1.4360±0.6551.1.8633±0.0979,1.9266±0.1724 in HaCaT cells 12 hours after irradiation to NB-UVB of 100,200 and 400 mJ/cm2,respectively,significantly higher than that in unirradiated cells(0.6000±0.1276,all P<0.05).Also,increased Gadd45α/β-actin protein ratio was noted in HaCaT cells 12 hours after irradiation to NB-UVB of 100,200 and 400 mJ/cm2 compared with unirradiated cells (0.0773±0.0005,0.1936±0.0015,0.2373±0.0015 vs.0.0290±0.0010,all P<0.05).NB-UVB inhibited the proliferation of HaCaT cells in a time-and dose-dependent manner.Flow cytometry showed that irradiated HaCaT cells were blocked in G2 phase of the cell cycle.and the percentage of HaCaT cells in G2 phase was 13.53%±1.03%,17.77%±2.25%,30.03%±4.29%afler exposure to NB-UVB of 100,200 and 400 mJ/cm2,respectively,compared to 9.24%±0.97%in unirradiated cells (all P<0.05).Conclusions The expression of Gadd45α is increased in HaCaT cells after exposure to NB-UVB,and Gadd45α may be involved in the NB-UVB-induced suprression of cell proliferation of and cell cycle arrest in HaCaT cells.