Objective:To establish the quality standard for infantile bronchitis traditional Chinese medicine lotion. Methods:TLC was used for the qualitative identification of Ephedra herba, Paeonia lactiflora Pall. , Poria cocos, Fructus aurantii and Epimedium brev-icornu Maxim. . HPLC was used for the content determination of ephedrine hydrochloride, pseudoephedrine hydrochloride and paeoni-florin. The chromatography conditions for ephedrine hydrochloride and pseudoephedrine hydrochloride were as follows: an AElichrom Pdar-Phenyl Polyphenyl ether-bonded phenyl-bonded silica column (250 mm × 4. 6 mm, 5μm) was used, the mobile phase was aceto-nitrile-0. 2% phosphoric acid solution (1 :99), the flow rate was 1. 0ml·min-1, the detection wavelength was 210 nm, the column temperature was 30℃ and the injection volume was 10 μl. The chromatography conditions for paeoniflorin were as follows:an Inertsil ODS-3 C18 column (250 mm × 4. 6 mm, 5 μm) was used, the mobile phase was acetonitrile-0. 1% phosphoric acid solution (15:85), the flow rate was 1. 0 ml·min-1, the detection wavelength was 230 nm, the column temperature was 30℃ and the injection vol-ume was 10 μl. Results:The TLC results of Ephedra herba, Paeonia lactiflora Pall. , Poria cocos, Fructus aurantii and Epimedium brevicornu Maxim. showed clear spots with good resolution. Ephedrine hydrochloride, pseudoephedrine hydrochloride and paeoniflorin had a good linear relationship within the range of 10.11-101.10 μg·ml-1(r=0.9996), 10.08-100.80 μg·ml-1(r=0.9991) and 20.50-102.5 μg·ml-1(r =0.9996), respectively. The average recovery was 98.80% (RSD =1.87%, n =6), 98.77%(RSD=1. 72%, n=6) and 99. 57% (RSD=1. 56%, n=6), respectively. Conclusion: The established quality standard can be used for the quality control of infantile bronchitis traditional Chinese medicine lotion.

Aim To investigate the effect of dehydroepiandrosterone(DHEA)on the levels of NR2B and GBR1 expressed in primary cultured rat cerebral cortical neurons.Methods Primary cultured rat cerebral cortical neurons were treated with DHEA of different concentrations (1,10,100 μmol·L~(-1))and the expression of amino acids receptor subunit NR2B and GBR1 were detected by immunocytochemistry.Results Compared with control group,the expression intensity of NR2B increased by 15.6%,19.9% and 49.4% after DHEA-L,DHEA-M and DHEA-H treatment(P<0.05 or P<0.01);the expression intensity of GBR1 increased by 14.5% and 58.5% after DHEA-M and DHEA-H treatment(P<0.05 or P<0.01).Conclusion DHEA can enhance the expression of neuron receptor subunit NR2B and GBR1.

Objective To investigate the effects of morphine dependence and withdrawal on neurosteroids and amino acid transmitters of rat amygdala. Methods Morphine dependence was induced by pretreatment with increasing doses of morphine for 7 days. Withdrawal was precipitated by naloxone (2mg/kg). Withdrawal syndromes were observed and scored. After decapitation, amygdala was dissected out. Nomadic and conjugated neurosteroids were extracted using liquid-liquid extraction and solid phase extraction. Concentrations of neurosteroids including dehydroepiandrosterone (DHEA), pregnenolone (PREG), allopregnanolone (AP), dehydroepiandrosterone sulfate (DHEAS) and pregnenolone sulfate (PREGS) were detected with HPLC-MS. Concentrations of glycine (GLY), glutamate (GLU) and gamma-aminobutyric acid (GABA) were quantitated by HPLC-ECD with pre-column OPA derivatization. Results Compared with saline control, the DHEA level in rat amygdala of morphine dependent group decreased by 33% (P＜0.01). Compared with naloxone control, the PREG and AP levels in rat amygdala of morphine withdrawal group increased by 45% (P＜0.05) and 42% (P＜0.05) respectively; the GABA level decreased by 18% (P＜0.01). Compared with morphine dependent group, the PREG and PREGS levels in rat amygdala of morphine withdrawal group increased by 60% and 40% respectively (P＜0.05); the glycine level decreased by 14% (P＜0.05). Conclusion The DHEA in rat amygdala may play a role in the development of morphine dependence but not involved in the manifestation of withdrawal symptoms. Other neurosteroids (including PREG, AP and PREGS) in rat amygdala seem to be involved in withdrawal but not in dependence. The synthesis and release of inhibitory amino acids in amygdala were depressed when withdrawal was precipitated by naloxone. The results suggest that different changes of neurosteroids and amino acids exist in stages of morphine dependence and withdrawal.

Aim To investigate the effect of progesterone on the levels of glutamate and ?-aminobutyric acid released from primary cultured rat cerebral cortical neurons.Methods Primary cultured rat cerebral cortical neurons were treated with PROG(10 ?mol?L-1) and the concentrations of amino acid in cell culture media at different time(0.5,1,1.5,2,24,36,48,72 h) were measured by OPA-mercaptoethanol precolumn derivatization technique and HPLC-FLD.Results Compared with control group,PROG treatment significantly reduced the levels of GLU at the time of 1,1.5,2,24,36,48,72 h(P

Objective: To investigate the effect of morphine dependence and withdrawal on the levels of neurosteroids in hippocampus of male rat.Methods: Rats were given (ip) increasing doses of morphine to form morphine physical dependence, withdrawal syndromes were precipitated by naloxone. The conditioned place preference (CPP) was used to establish morphine psychological dependence. The concentrations of dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), pregnenolone (PREG), pregnenolone sulfate (PREGS), and allopregnanolone (AP) in rat hippocampus and plasma were quantified by liquid chromatography-mass spectrometry. Results:The rat model of morphine physical and psychological dependence were successfully established by ip increasing doses of morphine for 7 days and 5mg?kg~ -1 morphine for 10 days respectively. Compared with saline control group, morphine physical dependence increased DHEA and PREG contents in rat hippocampus (0.88?0.19/0.67?0.17,t=2.52,10.94?2.02/7.53?2.64,t=3.24,P

Aim To investigate the effects of progesterone on morphine rewarding effect and levels of endogenous opioid peptides in hypothalamus of rat brain. Methods Conditioned place preference(CPP)test was used to investigate the morphine rewarding effect, and radioimmunoassay (RIA) was established to deter-mine levels of ?-endorphin(?-EP), leu-enkephalin(L-EK) and dynorphin A(DynA) in hypothalamus of rat brain.Results Compared with NS control group, 5 mg?kg-1 morphine successfully induced the formation of CPP(P

Aim To detect the effect of morphine dependence and withdrawal on the levels of neurosteroids and amino acid neurotransmitters in nucleus accumbens in rat morphine dependent model. Methods Nucleus accumbens was dissected out from morphine dependent and naloxone precipitated withdrawal rats. The contents of neurosteroids including dehydroepiandrosterone, pregnenolone, allopregnanolone, dehydroepiandrosterone sulfate and pregnenolone sulfate were detected with liquid chromatography-negative atmospheric pressure with ionization mass spectrometry(LC-MS). The contents of glycine, glutamate and ?-aminobutyric acid were quantitated by HPLC-ECD with precolumn derivatization. Results Compared with saline group,in nucleus accumbens of morphine withdrawal rats, the level of dehydroepiandrosterone sulfate (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) .

Objective To investigate the effect of propofol exposure on neuroapoptosis in pri-mary cultured cortical neurons and its mechanisms.Methods Cortical neurons were primarily cultured for seven days,then divided into two groups:control group (treated with equal volume of 20% in-tralipid),propofol-treated group (treated with 500 μmol/L propofol).The neurons were treated for 12 h.The neuron viability was determined by MTT.Neuroapoptosis was identified by Hoechest 33 258 dying.Mitochondrial membrane potential was measured by the fluorescent dye rhodamine 123 (Rh123).Western blot was performed to detect the level of cyt-c and cleaved-caspase-3.Results Neu-rons survival rate (54.4%±6.4%)in the propofol group was significantly lower than that of control group (99.8% ± 4.1%) (P < 0.05 ), the rate of neuronal apoptosis (46.5% ± 5.3%) was significantly higher than that of control group (7.2%±0.9%)(P <0.05),mitochondrial membrane potential (59.6%±4.3%)was significantly lower than that of the control group (99.9% ± 5.7%) (P <0.05 ),cyt-C protein level (0.38 ± 0.03 )was significantly higher than that of control group (0.1 5±0.02)(P < 0.05 ),level of cleaved-caspase-3 protein level (0.46 ± 0.04)was significantly higher than that of control group (0.13±0.02)(P <0.05).Conclusion Propofol induces neuroapo-tosis in primary cultured cortical neurons,which is associated with the decreased level of MPP and the increase levels of cyt-c and cleaved-caspase-3.

Objective Dexmedetomidine is known to have a neuroprotective effect.The aim of this study was to investigate the effects of dexmedetomidine on ketamine-induced apoptosis of primarily cultured cortical neurons and its action mechanisms. Methods Rat cortical neurons were primarily cultured for 7 days and treated with ketamine (100μmol/L) and different concentrations of dexmedetomi-dine (0.001, 0.01, 0.1, and 1 μmol/L) for 24 hours, followed by measurement of the viability of the neurons by MTT assay.The neurons were divided into four groups:vehicle control, ketamine ( trea-ted with 100 μmol/L ketamine), dexmedetomidine+ketamine (DD+K, treated with 0.1 μmol/L DD and 100 μmol/L ketamine), and LY294002 ( treated with 0.1 μmol/L DD, 100 μmol/L ketamine, and 10 μmol/L LY294002) .After 24 hours of treatment, the apoptosis rate of the neurons was determined by Hoechst33258 staining, and the expressions of pAkt and cleaved-caspase-3 in the neu-rons detected by Western blot. Results The apoptosis rate of neurons was dramatically increased in the LY294002 and ketamine groups in comparison with the vehicle control and DD+K groups ([36.8 ±4.4] and [43.4 ±4.5]%vs [7.5 ±1.1] and [16.4 ± 3.6]%, P<0.01), the pAkt level remarkably decreased (0.26 ±0.02 and 0.15 ±0.01 vs 0.61 ±0.05 and 0.50 ±0.04, P<0.01), and the expression of cleaved caspase-3 significantly upregulated in the former two as compared with the latter two groups (0.40 ±0.02 and 0.65 ±0.03 vs 0.10 ±0.02 and 0.12 ±0.01, P<0.01). Conclusion Dexmedetomidine exerts a neuroprotec-tive effect against ketamine-induced apoptosis of neurons by activating the PI3K-Akt signaling pathway.