Objective To investigate the effects of different doses of intrathecal magnesium on bone cancer pain (BCP) in mice.Methods Two hundred and eighty-eight male C3H/HeJ mice,aged 8-10 weeks,weighing 18-22 g,were randomly divided into 6 groups (n =48 each):control group (group C) ; sham operation group (group S) ; BCP + artificial cerebro-spinal fluid (aCSF) 5 μl group (group BCP) ; BCP + MgSO4 14.4 μg group (group M1 ) ; BCP + MgSO4 43.2 μg group (group M2 ) and BCP + MgSO4 86.4 μg group (group M3 ).BCP was produced by injecting fibrosarcoma cells of bone into the medullary cavity of right femur.Intrathecal catheter was placed in the 4 BCP groups.The aCSF 5 μl or MgSO4 14.4μg/5 μl,43.2 μg/5 μl,or 86.4μg/5 μl was injected intrathecally on 14th day after inoculation of tumor cells.The paw withdrawal threshold to mechanical stimuli (PWMT) and paw withdrawal lateney to thermal stimuli (PWTL) were measured at 0.5 h before administration (T0 ) and at 0.5,2,4 and 8 h after administration (T1-4).Eight animals chosen from each group at T0-4 were sacrificed,and L4-5 segment of the spinal cord was removed for determination of NR2B expression (by immuno-flurorescence) in the spinal cord.Results PWMT and PWTL were significantly decreased at T0-4,and NR2B expression was significantly up-regulated at T0-4 in groups BCP,M1,M2,M3 compared with groups C and S ( P ＜0.05).Compared with group BCP,PWMT and PWTL were significandy increased at T1-3,and NR2B expression was significantly down-regulated at T1-3 in groups M2 and M3 ( P ＜ 0.05).Compared with group M2,PWMT and PWTL were significantly increased at T1-3,and NR2B expression was significantly down-regulated at T1-3 in group M3 ( P ＜ 0.05).Conclusion Intrathecal magnesium can reduce BCP in a dose-dependent manner in mice.

Objective To investigate the effect of intraperitoneal injection of thalidomide on pain behaviors in a mouse model of bone cancer pain. Methods 36 male C3H/HeJ mice were divided randomly into tumor group (n= 18) and sham group (n= 18) ,six mice from each group were chosen to examine the time course of changes in behavior after tumor cells inoculated to the bone. 2 × 105 osteosarcoma NCTC 2472 cells were implanted into the intramedullary space of the right femurs of mice to induce ongoing bone cancer related pain behaviors. The sham group was inoculated by α-MEM without any cells. On the day before inoculation,the tumor mice were divided randomly into tumor + thalidomide group and tumor + vehicle group. The sham group mice were further divided randomly into sham + thalidomide group and sham + vehicle group. Pain ethology indexes such as paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) were observed on 1 d before inoculation and on 3 d ,5 d ,7 d, 10 d, 14 d after inoculation. Results ( 1 ) At day 7 after the operation, compared with sham mice ( 1. 70 ± 0. 33 ) g, PWMT of tumor mice decreased to ( 1.07 ± 0. 30) g (P ＜ 0. 05 ). At day 10, PWTL shortened to ( 12.60 ± 1.69 ) s (P ＜ 0. 05 ) compared with sham mice ( 17.70 ± 1.54 ) s. And the pain behaviors of tumor mice were aggravated along with the development of cancer pain. (2) At day 7 after the operation, compared with tumor + vehicle group ( 1. 07 ± 0.39 ) g, PWMT of tumor + thalidomide group increased to ( 1. 53 ± 0. 39 ) g (P ＜0.05). At day 10, PWTL extended to ( 16.48 ± 1.13 ) s compared with sham mice ( 12.64 ± 1. 56) s (P ＜0. 05 ). Conclusion Intraperitoneal injection of thalidomide can efficiently relieve mechanical hyperalgia and thermal hyperalgia in a mouse model of bone cancer pain.

ObjectiveTo investigate the effect of intrathecal injection of magnesium sulfate ( MgSO4 ) on pain behavior in mouse with bone cancer pain.Methods56 male 8-10 week old C3H/HeJ mice weighing 18-22 g were divided randomly into 7 groups ( n =8 ):sham group (S group),control group (C group) and MgSO4 plus morphine treat groups( T1-T5 group).Croup C and T mice were induced bone cancer pain models by intra-rightfemur inoculation of osteolytic NCTC2472 cells while group S were injected of only α-MEM.On the 14d after inoculation,group S and C received intrathecal injection of artificial cerebrospinal fluid 5 μl,while group T1-T5 received intrathecal injection of MgSO4 14.4 μg,43.2 μg,86.4 μg,morphine 0.36 μg,MgSO4 14.4 μg-morphine 0.36 μg,which were dissolved in 5 μl artificial cerebrospinal fluid.Micc received pain behavior tests including quantification of spontaneous flinches,paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) at 0.5h before and 0.5h,2h,4h,gh after administration.ResultsTreatment with MgSO4 (14.4 μg),morphine (0.36 μg) have no effect on bone cancer pain,while treatment with MgSO4 (43.2 μg,86.4 μg)can dose-dependently reverse quantification of spontaneous flinches,mechanical allodynia and thermal hypcralgesia which were induced by inoculation as well as MgSO4 14.4 μg-morphine 0.36 μg.At 0.5 h after administration,the quantification of spontancous flinches of the three groups( ( 10.08 ± 1.66),(7.35 ± 1.36),( 10.54 ± 1.32 ) ) were decrcased when compared with control group ( 13.05 ± 2.06 ),PWMT ( (0.81 ± 0.22 ) g; ( 1.33 ± 0.19)g; (0.93 ±0.26)g),PWTL( (10.57 ±1.53)s; (13.12 ±1.71)s; (11.46 ±1.83)s) were increased when compared with control group ( (0.42 ± 0.23 ) g,( 8.87 ± 1.27 ) s) (P ＜ 0.05 ).The effect reached maximum level at 2h,lasted for at least 4h and disappeared at 8h.ConclusionIntrathecal injection MgSO4 can effectively attenuate bone cancer pain dose-dependently.At the same time MgSO4 can amplify the analgesic effect of subliminal morphine.

Objective To investigate the effects of continuously intrathecal injection rapamycin on neuro pathic pain behaviors in mice.Methods 48 male adult C57/BL6 mice received intrathecal catheter implantation successfully and without motor dysfunction and serious weight loss,were choosed and randomly divided into shamoperation group ( sham,n =24) and chronic constriction injury model group ( CCI,n =24 ).After operation,each group randomly divided into 3 group again.Group I did nothing,group Ⅱ intrathecally injected rapamycin 1 μg/5μlon day 1 to 6 after operation,group Ⅲ intrathecally injected 20％ DMSO 5μl on the same time ( sham,CCI,sham +R,sham + V,CCI + R,CCI + V,n =8 ).Bilateral mechanical paw withdrawal threshold (WMT) and thermal paw withdrawal latency(TWL) were tested on day 1 before CCI and day 1,3,5,7,10,14,17,21,28 after operation.Results Compared with sham group,both WMT and TWL (7d,MWT:( 1.02 ±0.12)g vs (0.42 ±0.12)g,F=51.01,P＜0.05;TWL:(7.03 ±0.71 )s vs (3.26 ±0.66)s,F=38.27,P＜0.05) were significantly decreased after CCI on the ipsilateral side.When intrathecally injected Rapamycin 1 μg/5 μl on day 1 ～6 after CCI,the mechanical allodynia relieved obviously ( 7 d,M WT:( 0.42 ± 0.18 ) g vs ( 0.86 ± 0.25 ) g,F =6.56,P ＜ 0.05 ),and at least continued to 10 d.On the contrary,the effects of rapamycin on thermal hyperalgesia just showed a trend of inhibition,there was no statistic meaning.In addition,the sham group and contralateral pain behaviors did not change (P＞ 0.05 ).Conclusion Rapamycin can relieve the neuropathic pain behaviors in mice after CCI,mainly the mechanical allodynia,but not thermal hyperalgesia.

Objective:To explore the change characteristics of amino acid levels in neonates, so as to provide theoretical basis for accurate clinical interpretation.Methods:By preliminary screening and diagnosis from 32 855 newborns, 32 843 samples were collected using tandem mass spectrometry to inherited metabolic disease (IMD) scree-ning in Sichuan Province from January to December 2018.Afterwards, according to gestational age (1 363 premature infants, 31 468 full-term infants and 12 overdue infants), blood collection time (3-7 days old, 3 095 cases, 8-28 days old, 1 637 cases, and more than 28 days old, 248 cases) and season (7 737 cases in the first quarter, 11 428 cases in the second quarter, 5 482 cases in the third quarter, and 8 196 cases in the fourth quarter), neonates were divided into different study groups.The difference of amino acid level in each group was compared, and the correlation between various influencing factors and metabolic index was analyzed.Results:(1) The distribution of 11 amino acids [alanine(ALA), arginine(ARG), citrulline(CIT), glycine(GLY), leucine+ isoleucine+ hydroxyproline (LEU+ ILE+ PRO-OH), methionine(MET), ornithine(ORN), phenylalanine(PHE), proline(PRO), tyrosine(TYR), and valine(VAL)] in neonates showed non-normally distribution.(2)The distribution of 11 amino acids in different gestational age were tested by nonparametric test, except for PHE( H=0.61, P>0.05)and TYR( H=2.02, P>0.05), and other indicators were significantly different [ALA( H=187.11, P<0.05), ARG( H=23.60, P<0.05), CIT( H=22.90, P<0.05), GLY( H=85.18, P<0.05), LEU( H=56.42, P<0.05), MET( H=18.74, P<0.05), ORN( H=129.27, P<0.05), PRO( H=344.40, P<0.05), and VAL( H=272.92, P<0.05)]. (3) The distribution of 11 amino acids in different blood collection time were significantly different [ALA( H=65.19, P<0.05), ARG( H=404.48, P<0.05), CIT( H=502.13, P<0.05), GLY( H=1 719.44, P<0.05), LEU( H=396.41, P<0.05), MET( H=199.39, P<0.05), ORN( H=31.26, P<0.05), PHE( H=325.49, P<0.05), PRO( H=70.09, P<0.05), TYR( H=159.29, P<0.05), and VAL( H=102.52, P<0.05)]. (4) The distribution of 11 amino acids in different birth seasons were significantly different [ALA( H=401.37, P<0.05), ARG( H=3 229.03, P<0.05), CIT( H=65.45, P<0.05), GLY( H=597.82, P<0.05), LEU( H=1 120.42, P<0.05), MET( H=10 515.18, P<0.05), ORN( H=1 275.23, P<0.05), PHE( H=2 260.17, P<0.05), PRO( H=319.57, P<0.05), TYR( H=884.37, P<0.05), and VAL( H=1 824.49, P<0.05)]. Conclusion:According to different gestational age, season and blood collection time, the metabolism of amino acids in neonates was different.When using tandem mass spectrometry for detection, appropriate interpretation criteria should be selected based on different conditions.

Abstract By reviewing the advocacy framework and implementation path of global adolescent health promotion, summarizing the main health problems of adolescents worldwide and current status of adolescent health care services in China, an ecological model of adolescent health care was concluded according to the demands of adolescents towards health care services. A comprehensive intervention strategy, ie. multi sector cooperation, community participation and integration of hospital, school, community and family, etc, has been put forward to promote the health and sustainable development of adolescents.

Objective To evaluate the effects of artemisinin on proliferation,cell cycle and apoptosis of gallbladder cancer cells.Methods Gallbladder carcinoma cell lines (GBC-SD and NOZ) were cultured in vitro.The effects of artemisinin in different concentration on proliferation of the two cell lines in vitro were examined using MTT assay.The cell cycle distribution of GBC-SD and NOZ cells 24 h after treatments with artemisinin(20 μ mol/L) were examined using flow cytometry.The apoptosis of GBC-SD and NOZ cells 24 h after treatments with artemisinin (20 μ mol/L) were examined using Annexin V/PI staining.The expressions of p-ERK1/2,CDK4,cyclin D1,p16,cytochrome C and caspase-3 were examined by Western blot assay.t-test and one way ANOVA were used to evaluate the differences between two groups and more than two groups,respectively.Results The cell proliferation was significantly inhibited by artemisinin,the ICS0 of artemisinin against GBC-SD and NOZ cells were 14.05 μmol/L and 12.42 μmol/L,respectively.Artemisinin induced cycle arrest,and G1 population of GBC-SD and NOZ cells increased to 74.60％ and 78.86％.Cell apoptosis and apoptotic population of GBC-SD and NOZ cells were increased to 15.67％ and 16.51％ after dealt with artemisinin,respectively.In addition,expression of pl6 was increased,and expressions of p-ERK1/2,CDK4 and cyclin D1 were down-regulated by artemisinin(all P ＜0.05).Cytochrome C was released from mitochondria to cytoplasm leading to the activation of caspase-3 and PARP after dealt with artemisinin (P ＜ 0.05).Conclusion The inhibition effect of artemisinin on the proliferation gallbladder cancer cells is accompanied by down-regulation of ERK1/2 signaling pathway,G1 phase arrest and triggering caspase-3-mediate apoptosis.

Objective To evaluate the effects of artemisinin on proliferation,cell cycle and apoptosis of gallbladder cancer cells.Methods Gallbladder carcinoma cell lines (GBC-SD and NOZ) were cultured in vitro.The effects of artemisinin in different concentration on proliferation of the two cell lines in vitro were examined using MTT assay.The cell cycle distribution of GBC-SD and NOZ cells 24 h after treatments with artemisinin(20 μ mol/L) were examined using flow cytometry.The apoptosis of GBC-SD and NOZ cells 24 h after treatments with artemisinin (20 μ mol/L) were examined using Annexin V/PI staining.The expressions of p-ERK1/2,CDK4,cyclin D1,p16,cytochrome C and caspase-3 were examined by Western blot assay.t-test and one way ANOVA were used to evaluate the differences between two groups and more than two groups,respectively.Results The cell proliferation was significantly inhibited by artemisinin,the ICS0 of artemisinin against GBC-SD and NOZ cells were 14.05 μmol/L and 12.42 μmol/L,respectively.Artemisinin induced cycle arrest,and G1 population of GBC-SD and NOZ cells increased to 74.60％ and 78.86％.Cell apoptosis and apoptotic population of GBC-SD and NOZ cells were increased to 15.67％ and 16.51％ after dealt with artemisinin,respectively.In addition,expression of pl6 was increased,and expressions of p-ERK1/2,CDK4 and cyclin D1 were down-regulated by artemisinin(all P ＜0.05).Cytochrome C was released from mitochondria to cytoplasm leading to the activation of caspase-3 and PARP after dealt with artemisinin (P ＜ 0.05).Conclusion The inhibition effect of artemisinin on the proliferation gallbladder cancer cells is accompanied by down-regulation of ERK1/2 signaling pathway,G1 phase arrest and triggering caspase-3-mediate apoptosis.