1.Roles of ERK-CREB signaling pathway in spinal cord in naloxone-induced withdrawal response in morphine-dependent rats
Chinese Journal of Anesthesiology 2012;32(3):307-310
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.
2.Role of NO and ERK signaling pathways in spinal cord in naloxone-induced withdrawal response in morphine-dependent rats
Chinese Journal of Anesthesiology 2011;31(8):938-942
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.
3.Effectiveness of Different Indoor Air Disinfection Methods
Journal of Environment and Health 1992;0(04):-
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.
4.Drug delivery of CPC/cisplatin complex in vitro and its ability to repair bone defect and eliminate tumor in vivo
Yanning LIU ; Miao LIU ; Pengyu REN
Journal of Central South University(Medical Sciences) 2009;34(10):991-997
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.
5.Perioperative changes of erythrocytes hexokinase, phosphofructokinase and glucose-6-phasphate dehydrogenase activities in patients undergoing upper abdominal surgery
Youli HU ; Yanning QIAN ; Cunming LIU
Chinese Journal of Anesthesiology 1995;0(10):-
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
6.The effect of cholecystectomy under epidural block on glucose transport-4 mRNA expression in skeletal muscle
Yanning QIAN ; Youli HU ; Cunming LIU
Chinese Journal of Anesthesiology 1996;0(07):-
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.
7.Human umbilical cord mesenchymal stem cells protects against Aβ-induced impairment partly through up-regulation of expression of neurotrophins
Sha LIU ; Ming WU ; Bingqi SHI ; Zengjuan LIU ; Yanning HOU
Chinese Pharmacological Bulletin 2016;32(7):980-985
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) .
8.Influence of different intervention methods on the second stage of labor of primipara
Xiuhong LONG ; Yanning HU ; Guifeng LI ; Qingyan LIU ; Jing LIANG
Chinese Journal of Practical Nursing 2011;27(7):4-6
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.
9.Differentiation efficiency of human umbilical cord mesenchymal stem cells into hepatocytes under two kinds of liver homogenate supernatants: a comparative study
Cheng YAN ; Gai XUE ; Liying WU ; Jianfang LIU ; Yanning HOU
Chinese Journal of Tissue Engineering Research 2015;19(19):2993-2998
BACKGROUND:Previous studies have demonstrated that normal rat liver homogenate supernatant can induce human umbilical cord mesenchymal stem cels to differentiate into hepatocyte-like cels with partial hepatocyte functions. However, whether fibrotic liver homogenate supernatant can work or how the inducing effect is remains unclear. OBJECTIVE:To investigate the differentiation potential of human umbilical cord mesenchymal stem cels into hepatocytes under the normal liver and fibrotic liver microenvironment in vitro. METHODS:Liver fibrosis was induced in the SD rats by repeated intraperitoneal injections of 3% thioacetamide at a dose of 200 mg/kg body mass, twice a week for 4 weeks, and then fibrotic liver tissues and normal liver tissues were used to prepare liver homogenate supernatants. Passage 3 human umbilical cord mesenchymal stem cels were used and divided into standard control group (cels were cultured in DMEM/F12 with 10% fetal bovine serum), fibrotic liver homogenate supernatants group (cels were cultured in DMEM/F12 with 10% fetal bovine serum and 50 g/L fibrotic liver homogenate supernatants), normal liver homogenate supernatants group (cels were cultured in DMEM/F12 with 10% fetal bovine serum and 100 g/L normal liver homogenate supernatants). The morphological changes of the cels in each group were recorded under inverted microscope; the protein levels of CK18, AFP, CYP3A4, CYP2E1, CYP2D6 and TPH2 were evaluated using western blot assay. Furthermore, the concentration of albumin in the cels was measured. RESULTS AND CONCLUSION:After a 7-day inducement, the stem cels in liver homogenate supernatants groups lost their fusiform shape and changed into hepatocyte-like cels with the morphous of round shape. Compared with the standard control group, the hepatocyte-like cels in the two liver homogenate supernatants groups exhibited human hepatocyte biomarkers, CK18 and AFP. The standard control group cels could express a little amount of CYP2E1, while cels in the two liver homogenate supernatants groups could express CYP3A4, CYP2E1, CYP2D6, TPH2. Compared with the standard control group, the expression level of CYP2E1 in the two liver homogenate supernatants groups increased significantly (P < 0.01), and however, the relative levels of CYP3A4, CYP2E1, CYP2D6, TPH2 in the two liver homogenate supernatants groups showed no statistical significance (P > 0.05). At the same time, compared with the standard control group, the concentration of albumin in the two liver homogenate supernatants groups markedly increased (P < 0.01), but there was no difference between the two liver homogenate supernatants groups (P > 0.05). Experimental findings demonstrated that both of normal liver tissue and fibrotic liver tissue microenvironments could induce human umbilical cord mesenchymal stem cels to differentiate into hepatocyte-like cels. To achieve the same effect, compared with normal liver tissue, fibrotic liver tissue required lower concentrations, suggesting that fibrotic liver tissue microenvironment may be more conducive to differentiation of umbilical cord mesenchymal stem cels into hepatocytes.
10.Application effect of massaging Changqiang acupuncture point during the second labor course
Xiuhong LONG ; Yanning HU ; Guifeng LI ; Qingyan LIU
Chinese Journal of Practical Nursing 2010;26(20):15-16
Objective To investigate the effect of massaging Changqiang acupuncture point during the second labor course of primiparas. Methods 124 primiparas were randomized into the observation group(63 cases)and the control group (61 cases). In the observation group, after entering the second stage of labor, Changqiang point was massaged upwards by using index finger and middle finger, and at the same time primiparas were guided to hold breath to coordinate with uterine contraction to make effort to the anus point which was massaged. While the control group were directed with routine way of correctly using abdominal pressure during process of uterine contraction in the second stage of labor. Results The lasting time of uterine contraction was significantly longer in the observation group than in the control group, the time of second stage of labor was shorter, and the amount of postpartum hemorrhage in the observation group was less than those in the control group. Conclusions Application of massaging Changqiang acupuncture point could prolonge the time of uterine contraction, advance the progress of second stage of labor, and decreased the amount of postpartum hemorrhage.