Aim This study aimed to explore the effects of intrathecal injection of nitric oxide synthase inhibitors(L-NAME,7-NI) on morphine withdrawal response and the the spinal p-ERK expression.Methods Rats were divided into 5 groups: control group,dependence group,withdrawal group, L-NAME group(L-NAME,it) and 7-NI group(7-NI,it).Morphine withdrawal score,touch evoked agitation scores(TEA scores),immunohistochemical and western-blotting technique were used to evaluate morphine withdrawal response and the expression of Fos and p-ERK in the spinal cord,respectively.Results Intrathecal injection of non-specific NOS inhibitor L-NAME,nNOS inhibitor 7-NI significantly alleviated morphine withdrawal symptoms.Morphine withdrawal scores in the L-NAME(22.1?4.52) and 7-NI groups(16.2?3.99) were significantly lower than that of the withdrawal group(28.6?4.89)(P

BACKGROUND:In recent years, continuous femoral nerve block has been extensively used in total knee replacement, but there has a great dispute on the effect of analgesic mode in the clinic. OBJECTIVE:To compare the analgesic effects of continuous femoral nerve block and conventional intravenous patient-control ed analgesia in total knee arthroplasty. METHODS:A total of 50 cases treated with unilateral total knee arthroplasty in the Department of Orthopedics, Huaian First Hospital Affiliated to Nanjing Medical University from January 2014 to January 2015 were enrol ed in this study. Primary disease was osteoarthritis of the knee. They were randomly divided into two groups. Patients in the control group received intravenous patient-control ed analgesia, and those in the test group received continuous femoral nerve block analgesia. Postoperative pain Visual Analog Scale scores, adverse reactions and analgesic satisfaction score were compared between the two groups. RESULTS AND CONCLUSION:The Visual Analog Scale scores were significantly lower in the test group than in the control group at 4, 12, 24, 48 and 72 hours after surgery (P<0.05). The application rate of analgesic adjuvant drugs within 48-72 hours after surgery was significantly lower in the test group (4%) than in the control group (24%) (P<0.05). The incidence of complications was significantly lower in the test group (12%) than in the control group (48%) (P<0.05). The postoperative analgesic satisfaction rate was significantly higher in the test group (88%) than in the control group (60%) (P<0.05). Range of motion of the affected knee was significantly bigger in the test group than in the control group at 2, 3 and 4 days after replacement (P<0.05). Length of stay was significantly shorter in the test group than in the control group after surgery (P<0.05). These results suggest that multimodal continuous femoral nerve block analgesia has an ideal analgesic effect in total knee replacement. The incidence of adverse reactions is low. The analgesic satisfaction rate is high. Thus, it is an ideal analgesic method after total knee replacement.

Objective To investigate the feasibility of using PLGA loaded with SD rats' mesenchymal stem cells(rMSCs) transfected with green fluorescent protein (GFP) gene as scaffolds for combinations of molecular, cellular, and tissue-level treatments of spinal cord tissue engineering. Methods rMSCs infected with lentiviral vectors (lv-GFP) were seeded onto PLGA at 8000 cell/cm2, rMSCs-GFP grown under similar conditions on tissue culture plastic as control. The morphology of rMSC-GFP was examined by fluorescence microscopic. The activity of MSCs was detected by MTT assay everyday. Cell cycle analysis was performed after a 3-day culture on PLGA using flow cytometry. The rMSCs-GFP seeded on PLGA was identified by FITC-anti CD34,CD90 and PE-anti CD44, CD106,CD45,CDllb at the third day. Results Fluorescence microscopic examination revealed adherence of the cells to the PLGA surface within 24 h of initial plating. After 3 days, GFP cells were spindle shaped. The difference disappeared at 7 days when cells under both conditions had become confluent Cells proliferated at the same rate on the PLGA surface compared to tissue culture plastic. And cell's cycle was unaffected by the transduction process and seeded on PLGA. Cells maintained their stem cell phenotype as judged by expression of CD90, CD44, CD106 markers,and absence of the hematopoietic marker CD45, CD34. This demonstrated that the transduction and the PLGA surface were not adversely affecting the cells. Conclusions MSCs are a good candidate for spinal cord tissue engineering. Cells continued to express green fluorescent protein(GFP)on a long-term basis,and are compatible with polymer surfaces. Morphology,viability,and growth kinetics were maintained when cells were grown on a poly-lactic-co-glycolic acid(PLGA)polymer scaffold. Therefore,they could make further efforts for combinations of molecular, cellular,and tissue-level treatments of spinal cord tissue engineering.

Objective To evaluate the role of extracelluar signal-regulated kinase (ERK)-cyclic AMP response element binding protein (CREB) signaling pathway in the spinal cord in naloxone-induced withdrawal response in morphine-dependent rats.Methods Fifty male adult SD rats,aged 2 months,weighing 200-250 g,in which intrathecal catheters were successfully implanted without complications,were randomly divided into 5 groups (n =10 each):group control (group C); group morphine dependence (group MD); group morphine withdrawal (group MW); group U0126 (ERK signaling pathway blocker); group dimethyl sulfoxide (DMSO,solvent for U0126).Morphine dependence was induced by increasing doses of subcutaneous morphine for 6 days.The initial dose of morphine was 10 mg/kg twice a day and was increased by 10 mg/kg twice every other day until 50 mg/kg on 6th day in groups MD,MW,U0126 and DMSO.Morphine withdrawal response was induced by intraperitoneal naloxone 4 mg/kg at 4 h after last morphine administration in groups MW,U0126 and DMSO.U0126 150μg (in DMSO 10 μl) and DMSO 10 μl were administered intrathecally at 30 min before naloxone administration in groups U0126 and DMSO respectively.Morphine withdrawal response (0=no withdrawal response,3 =severe response)and touch evoked agitation (0 =no agitation,2 =severe agitation) were observed and scored during 1 h after naloxone administration.The animals were then sacrificed and the spinal cord was removed for determination of the expression of phosphorylated ERK (p-ERK) and phosphorylated CREB (p-CREB) by immuno-histochemistry and Western blot.Results Morphine withdrawal significantly up-regulated the p-ERK and p-CREB expression in group MW compared with group C ( P ＜ 0.05).Withdrawal response score and touch evoked agitation score were significantly increased in groups MW,U0126 and DMSO as compared with group MD ( P ＜ 0.05).U0126 pretreatment significantly attenuated naloxone-induced increase in withdrawal response score and touch evoked agitation score and down-regulated p-ERK and p-CREB expression in group U0126 as compared with group MW ( P ＜ 0.05).Conclusion ERK-CREB signaling pathway in the spinal cord is involved in morphine withdrawal response in morphine-dependent rats.

ObjectiveTo investigate the role of NO and extracellular signal-regulated kinase (ERK) signaling pathways in the spinal cord in naloxone-induced withdrawal response in morphine-dependent rats.Methods Ninety male adult SD rats weighing 200-250 g in which IT catheters were successfully implanted without complication were randomly divided into 9 groups (n ＝ 10 each):group control (group C); group morphine dependence (group MD); group morphine withdrawal (group MW); group N(G)-nitro-L-arginine methyl ester (eNOS inhibitor) (L-NAME group) ; group 7-nitroindazole (nNOS inhibitor) (group 7-Ni) ; group aminoguanidine(iNOS inhibitor) (group AG); group U0126 (ERK signaling pathway blocker); group cremophor (solvent for 7-Ni) and group DMSO (solvent for U0126).Morphine dependence was induced by increasing doses of subcutaneous morphine for 6 days.The initial dose of morphine was 10 mg/kg twice a day and was increased by 10 mg/kg twice every other day until 50 mg/kg on the 6th day in groups MD,MW,L-NAME,7-Ni,AG,U0126,cremophor and DMSO.Morphine withdrawal response was induced by intraperitoneal (IP) naloxone 4 mg/kg at 4 h after last morphine administration in groups MW,L-NAME,7-Ni,AG,U0126,cremophor and DMSO.L-NAME 400 μg,7-Ni 400 μ g,AG 400μg,U0126 150 μg,cremopher 10 μl and DMSO 10 μl were administered IT at 30 min before naloxone administration in groups L-NAME,7-Ni,AG,U0126,cremophor and DMSO respectively.Morphine withdrawal response (0 ＝ no withdrawal response,3 ＝ severe response) and touch evoked agitation (0 ＝ no agitation,2 ＝ severe agitation) were observed and scored during 1 h after naloxone administration.The animals were then sacrificed and the spinal cord was removed for determination of the expression of iNOS,nNOS and phosphor-ERK (p-ERK) by immunohisto-chemistry and Western blot.ResultsMorphine withdrawal significantly increased withdrawal response score and touch evoked agitation score in group MW as comparedwith group MD.L-NAME,7-Ni,AG and U0126 pretreatment significantly attenuated naloxone-induced increase in withdrawal response score and touch evoked agitation score in groups L-NAME,7-Ni,AG and U0126 as compared with group MW.Morphine withdrawal significantly up-regulated the nNOS and iNOS expression in group MW compared with groups C and MD.L-NAME,7-Ni and AG pretreatment significantly down-regulated p-ERK expression in groups L-NAME,7-Ni and AG as compared with group MW.ConclusionThe interaction between NO and ERK signaling pathways may be involved in morphine withdrawal response in morphine-dependent rats.

Objective To observe the curative effects of azithromycin combination with clindamycin on the treatment of children with mycoplasmal pneumonia. Methods 146 children were divided into control group and observation group randomly.Azithromycin was used to treat 69 children in the control group while azithromycin combination with clindamycin was used to treat 77 children in the observation group.The curative effects and adverse effects were observed. Results The total effective rate of control group was 81.2%,which was significantly lower than that in observation group(94.8%),(x2=6.59,P＜0.05). Conclusion Azithromycin combination with clindamycin was an effective and safety method to treat the children with mycoplasmal pneumonia.

Objective To study the effects of octreotide on the proliferation and apoptosis of hepatocellular carcinoma cells, and provide an experimental basis for clinical application. Methods The proliferation of hepatocellular carcinoma cells (HepG2) was assessed by MTT and growth curve respectively, the contents of AFP in the culture supernatant were determined by eletrochemiluminescence immunoassay, and apoptosis was detected by fluorescent staining, transmission electron microscopy and flow cytometry. Results The proliferation of HepG2 was inhibited significantly by octreotide with a dosage dependant manner(range from 0.005 to 80 ?g/ml, P

Purpose:T0 study the mechanism and the potentiality of calciumchannel blocker to prevent organal fibrogenisis. Materials and Mehods:The effects ofnifedipine(Nif)and nicardipine (Nic)on proliferation of human lung fibroblasts(HLF)and synthesis of coliagen and hyaluronic acid(HA)were determined by means of MTT,measuring the incorporation of 3H-proline and radioimmunoassay, respectively.Results: both Nif and Nic supressed HLF proliferation and collagen synthesis,as wellas decreased the production of HA in a concentration-dependent manner at l0-40?mol/L. In addition,there were no significant toxic actions on HLF. Conclusion: Nifand Nic might be hopeful antifibrotic drugs.

As an outward,voltage-dependent potassium channel,M type channel is crucial in the regulation of neuronal excitability;it is modulated by a variety of factors in vivo and its dysfunction often results in neuronal system diseases.Great efforts have been made to elucidate the mechanism underlying M channel modulation since its discovery decades ago.It is generally accepted that the Phospholipase C(PLC) signaling pathway plays a significant role in the M channel modulation.This review highlights the relationship between PLC signaling pathway and M channel modulation,as well as some recent progresses in the research of this field.

Receptor tyrosine kinase(RTK),a membrane receptor superfamily with intrinsic protein tyrosine kinase activity,has many members and complicated signal transduction pathways.Activation of RTKs can trigger a series of signal transduction pathways and play essential roles in modulating cell growth,proliferation,differentiation and metabolism through influencing gene transcription and expression.Activation of RTK can also rapidly modulate some cellular functions including the modulation of ion channels.Potassium channels play a critical role in stabilization of membrane potential and regulation of cellular excitability.This review highlights the rapid modulation of potassium channels by RTKs and reviews the recent progress in related research.