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 explore a simple, effective and rapid method for indoor air disinfection. Methods Three kinds of disinfection methods were carried out (method A: wiping surfaces of floor and walls of room with water and ultraviolet irradiation, method B: wiping surfaces of floor and walls with chloro-disinfectant and ultraviolet irradiation, method C: wiping surfaces with chloro-disinfectant and spraying chloro disinfectant containing chlorine) and the effectiveness of different disinfection methods was compared. Results The result showed that all of the three methods could decrease natural fallen bacteria count in isolated room air to up to the standard, and the disinfection effectiveness of method C was better than the method A and method B. Conclusion The result indicated that method C was an effective and simple method for indoor air disinfection.

Objective To explore the best mass ratio of calcium phosphate cement (CPC) / cisplatin complex filling and to repair bone defect caused by tumor resection. Methods Mixed-molding method was used to obtain cisplatin/calcium phosphate cement complex at 0, 0.1%, 0.2%, and 0.4% mass ratio. Drug concentration was determined by atomic absorption spectrophotometry. Bone defect of rabbits and osteosarcoma of rats were prepared. We implanted CPC and CPC/cisplatin complex to observe the repair of bone defect and the inhibition of tumor in vivo. Results CPC containing 0.1% ～ 0. 2 % cisplatin not only repaired the bone defect in rabbits but also eliminated osteosarcoma in rats. Conclusion CPC containing 0.1% ～ 0. 2 % cisplatin can repair bone defect and eliminate tumor without influencing the prosthetic precess.

Objective Surgical stress induces a series of endocrine and metabolic changes including glucose metabolism and insulin-resistance. The purpose of the present study was to investigate the changes in glucose transporter-4 (Glut-4) mRNA expression in skeletal muscle after cholecystectomy under epidural block. Methods One gram of the rectus abdominis muscle was taken while abdomen was being opened and closed in patients undergoing cholecystectomy under epidural block. Total RNA of the muscle cells was extracted by trizol one-step template. RT-PCR was used to determine the Glut-4 mRNA amplification products with ?-actin mRNA as an internal control. The Glut-4 mRNA expression was expressed by (desired gene/ ?-actin?100% . The plasma glucose and insulin levels were determined at the same time. Results Glut-4 mRNA expression was significantly reduced (P 0.05 ) . Conclusions The results indicated that the synthesis of Glut-4 is suppressed by surgical stress of cholecystectomy under epidural block leading to insulin resistance.

Objective To investigate the role of pentose phosphate pathway (PPP) after upper abdominal operationMethods Twenty-six patients undergoing upper abdominal surgery, were divided randomly into three groups: epidural block(EB,n=10), intravenous propofol anesthesia(PRO, n=10) and intravenous procaine balanced anesthesia(IPBA,n=6) The venous blood samples were collected to measure of erythrocytes hexokinase (HK), phosphofructokinase (PFK) and glucose-6-phasphate dehydrogenase (G-6PD), and activities, and plasma glucose before anesthesia induction, 60 min after incision,60 min after surgery, on the 1st and 2nd postoperative daysResults Compared with the values before anesthesia, the activities of erythrocyte HK and PFK decreased significantly on the 1st postoperative day in three groups (P

Aim To investigate the protective effect of human umbilical cord mesenchymal stem cells ( hUCM-SCs ) against Aβ-induced impairment in rats and the possible mechanism .Methods Male SD rats ( weight 210~230 g ) were divided randomly into five groups with ten in each:① control group ( con );② vehicle-control group ( sterile distilled water , v-con );③ hUC-MSCs-control group ( hUCMSCs-con );④ Aβinjury group ( injury );⑤ hUCMSCs treatment group ( hUCM-SCs) .Six-day Morris water maze behavioral task was employed to test the spatial learning and memory of the animals.Neuro-pathological evaluation under thionin stain was performed after behavioral task .The level of brain-derived neurotrophic factor ( BDNF ) and nerve growth factor ( NGF ) were tested through ELISA .Re-sults hUCMSCs enhanced the cognitive performance of Aβtreated rats, and reversed Aβ-induced cell loss in CA1 hippocampus.On the cellular level, hUCMSCs attenuated Aβinjection induced down-regulation of NGF and BDNF.Conclusion Administration of hUC-MSCs can reverse the behavioral and cellular impair-ment of Aβtreated rats, as well as the down-regulation of neurotrophins , thus exerting a neuronal protective effect, which provides a potential therapeutic strategy for disorders with learning and memory impairment , such as Alzheimer ’ s disease ( AD) .

BACKGROUND:Healthy human umbilical cord jel y is a mucous connective tissue without vessels and has been used in eye plastic operation because of its elasticity and toughness. It contains lysozyme, placental globulin, hyaluronic acid enzyme, AMP antibody and complement, and also contains a lot of mesenchymal stem cells, so it is not prone to infection and rejection. OBJECTIVE:To observe the histocompatibility and histopathological changes of human umbilical cord jel y as a tarsal substitute transplanted for eyelid reconstruction in guinea pigs. METHODS:The mucous connective tissue of healthy neonate umbilical cord jel y was made into tissue blocks at even thickness. Models of tarsal defects were established in guinea pigs, and then the mucous connective tissue of healthy neonate umbilical cord jel y was implanted. Samples of implanted materials were col ected for histological examination at 1, 2, 3 weeks postoperation under light microscope. RESULTS AND CONCLUSION:There were no obvious rejection, infection and eyelid deformation observed, the corneas of al the animals were clear, corneal epithelial shedding did not occur, and the eyelid could move normal y. One week after implantation, there was no infection and rejection on the conjunctiva and the incision of the eyelid, the eyelid could move normal y, and partial inflammatory cells were observed between the human umbilical cord jel y and the muscle of the eyelid with microscopy. Two weeks after implantation, there was no infection and rejection on the conjunctiva and the incision of the eyelid, the cornea was clear, the eyelid and eye could move normal y, and no symblepharon occurred;umbilical cord jel y showed the tendency of absorption, and the inflammatory cells were reduced at 2 weeks after implantation. Three weeks after implantation, the incision of the conjunctiva healed wel , the cornea was clear;there was no difference in the eyelid form and movement between the two eyes;the umbilical cord jel y was absorbed partial y, normal fibrous tissues formed and there were no inflammatory cells. With low immunogenicity, human umbilical cord jel y can guide the growth of new col agen and serve as an ideal tarsal substitute.

Objective To study the influence on primipara's second stage of labor with the methods of Changqiang-pressing and Vaginal-pressing. Methods 180 primipara in the second stage of labor were randomly divided into 3 groups of 60 each, in which Changqiang-pressing, Vaginal-pressing and general guidance (the control group) were used respectively. The duration of uterine contraction and second stage of labor, vaginal bleeding amount in 2 hours after delivery, mode of delivery, perineal injury and birth case would be evaluated. Results The results in both Changqiang-pressing group and Vaginal-pressing group were statistically significant when compared with the control group, but there was no significant difference between Changqiang-pressing group and Vaginal-pressing group. Conclusions Using Changqiang-pressing and Vaginal-pressing can promote the progress of the second stage of labor for primipara, reduce the rate of cesarean section and neonatal asphyxia, and improve the quality of obstetric department.

BACKGROUND:Preliminary experiments have demonstrated that rat liver homogenate supernatants can induce the morphological changes of human umbilical cord mesenchymal stem cells. However, little is known about whether the induced cells have some phenotypic and functional features of hepatocytes.
OBJECTIVE:To investigate whether human umbilical cord mesenchymal stem cells have some phenotypic and functional characteristic of hepatocytes after being induced by liver homogenate supernatants.
METHODS:Passage 3 human umbilical cord mesenchymal stem cells were used and divided into control group (cells were cultured in basic culture medium) and liver homogenate supernatant group (cells were cultured in liver homogenate supernatants for 3, 5, 7 days). Meanwhile, positive control group (QSG-7701 human liver celllines) and negative control group (simple liver homogenate supernatants) were set up. The protein and mRNA level of hepatocyte markers, alpha-fetoprotein, cytokeratin 18 and tryptophan 2,3-dioxygenase enzyme, were detected at different time points.
RESULTS AND CONCLUSION:After inducement, the stem cells of fusiform shape began to lose their sharp edges and progressively shrunk, and then they changed into hepatocyte-like cells with the morphous of triangle, polygon and anomalism shape. Compared with the control group, the protein and mRNA level of alpha-fetoprotein, cytokeratin 18 and tryptophan 2,3-dioxygenase enzyme significantly increased time dependently after inducement with liver homogenate supernatants (P<0.01). This study demonstrated that human umbilical cord mesenchymal stem cells are able to differentiate into hepatocyte-like cells in vitro that possess some functions of liver cells.