Objective To investigate the effects of icariin on the expressions of osteoprotegerin(OPG)and receptor activator of nuclear factor-kB ligand(RANKL)mRNA in rat osteoblasts cultured in vitro.Methods Calvarial osteoblasts were obtained from newborn SD rats aged＜24 h by trypsin-collagenase digestion method. After treatment with different concentrations of icariin(10~(-10)-10~(-4) mol/L)for 48 h,total RNA was isolated from osteoblasts.Expressions of OPG and RANKL mRNA were detected by semiquantitative RT-PCR.Results Icariin remarkably increased OPG mRNA levels and decreased RANKL mRNA levels in calvarial osteoblasts in a dose- dependent fasion with a maximum effect at a concentration of 10~(-7)mol/L(P＜0.05).Osteoblasts pre-treated with icariin prevented the inhibitory effect of dexamethasone on OPG mRNA(0.570?0.093 vs 1.083?0.081)and the stimulative effect on RANKL mRNA(1.079?0.050 vs 0.718?0.082)(both P＜0.05).Conclusion Icariin upregulates OPG expression and downregulates RANKL expression in osteoblasts,which may explain the beneficial role of icariin in preventing and treating osteoporosis.

Growth dynamics of PP, CN and ZS2 strains of Toxoplasma gondii isolated in China was studied and compared with that of RH strain. HeLa cells were used in this work. It took only 2 min for the organisms of RH strain to infect the HeLa cells when con-tacting with the cells. By contrast, the CN strain requires 5 min, the ZS2 and PP strains, 10 min.Tcxoplasma began its multiplication after a lag time of about 6 h in the HeLa cells. The mean generation time of the 4 strains was assessed by calculating the number of the parasites in the parasitophorous vacuoles at different incubation times and by the linear regression equation. The results showed that the mean generation time was 5.2 h for RH strain, 5.98 h for CN strain, 6.78 h for ZS2 strain and 7.69 h for PP strain.Among the 3 strains of Toxoplasma gondii isolated in China, CN strain was similar to RH strain in their infectivity and proliferation.

Objective Blood lactic acid(LA) and glucose(Glu) level are important parameters of anaerobic glycolysis and can be used to assess the severity of brain injury and cerebral metabolism. The purpose of this study was to evaluate effect of propofol on traumatic brain injury by measuring blood and CSF level of LA and Glu in addition to microscopic and NSE immunohistochemical examination of brain tissue. Methods Ninety New Zealand rabbits weighing 2.6-3.0 kg were used . Traumatic brain injury model was established according to Wang's method. Part Ⅰ . Twenty rabbits were divided into two groups of ten animal each. Blood and CSF samples were taken before and 4h, 24h, 48h, 72h and 1 week after trauma for determination of LA and Glu levels. Propofol group received propofol 30mg' kg-1?h-1 infusion for 30 min in addition to ketamine 1mg/kg before each collection of samples. PartⅡ . Seventy rabbits were divided into seven groups with ten animals in each group. Brain tissues were taken before and 24h, 72h, and 1 week after trauma for microscopic and NES immunohistochemical examination. Propofol group received infusion of 30 propofol mg kg-1 h-1 for 30 mm every day. In control group animals received same amount of normal saline. Results Blood and CSF levels of LA increased significantly after trauma in both groups but were significantly higher in control group than those in propofol group at corresponding intervals. Blood and CSF Glu levels decreased significantly in control group after trauma but in propofol group blood Glu level decreased only at 4h and 24h after trauma and CSF Glu level at 24h after trauma. There was significant difference in blood and CSF levels of Glu after trauma between the two groups. In both groups microscopic examination of brain tissue showed hemorrhage, degeneration, decrease in glial cells and vacuolization of some neuron in brain tissue of injured and surrounding areas at 24h after trauma, infiltration of neutrophils at 72h after trauma and cerebral interstitial edema and glial cell proliferation at 1 week after trauma. Neurons showed no NSE expression. In propofol group the above mentioned changes were relatively slight. Conclusions Propofol can significantly reduce blood and CSF LA levels after trauma and protect the animal from traumatic brain injury. Further studies are needed on the dosage and method of administration of propofol.

Objective To evaluate the effect of allicin alone or combined with SMZco on murine toxoplasmosis by aspecific, rapid, and sensitive PCR technique. Methods 147 mice were infected with 2?10~4 tachyzoites intraperitoneallyand divided into 5 groups at random. Each group was divided into two sub-groups except an untreated group. One sub-group was used to get samples for PCR test and the other for observing the survival duration. The therapeutic grouping wasas follows: group A, a combination of allicin and SMZco administered orally for 7 days and continued by allicin alone till 21days; group B, the combination administered for 14 days and continued with allicin till 21 days; group C, allicin alone for21 days; group D, SMZco alone for 7 days; group E, untreated control. The dosage was: SMZco 400 mg/(kg?d) and al-licin 35 mg/ (kg?d). PCR test was used to detect the parasites in samples of liver and blood from infected mice at 5, 10, 15,20, 25, 30, 40 and 50 days after infection. Results Parasites were eliminated in the blood because no signal was seen inall the blood samples except for samples from group C at day 5 after infection. From day 10 after infection until the end ofthe experiment, no amplification of DNA was seen in all the samples. As for liver samples, signals were clear at day 5 postinfection. From day 10 post infection till the day 50 post-infection, parasites were still detected, but the PCR products de-creased significantly than that of day 5 post-infection. Result showed that a combination of SMZco with allicin provided asignificant protection. SMZco alone was also effective, but allicin alone was not. Conclusion When SMZco is used incombination with allicin, a much higher efficacy is received in the treatment of acute murine toxoplasmosis.

To observe the effect of propofol on the change in neuron specific enolase(NSE) contents in the serum and cererospinal fluid(CSF) of rabbits suffering from brain injury, so as to explore the protective effect of propofol on the brain, 20 New Zealand rabbits were inflicted with brain injury and randomized into control group and propofol treatment group ( n =10 each). Samples of serum and CSF were collected before trauma and 4h, 24h, 48h, 72h and one week following trauma respectively. The contents of NSE in serum and CSF were measured by enzyme immunoassay (EIA) method. The results showed that the content of NSE in serum at 24h, 48h, 72h and one week, and that in CSF from 4h to one week after trauma in control group were elevated significantly as compared with those before trauma ( P

Objective To investigate the curative effect and care measures using subhypothermia treatment in patients with severe brain injury.Methods 70 cases of severe brain injury were randomly divided into treatment group(n=35) and control group (n=35),the treatment with subhypothermia and correlation care,control group treated with conventional treatment care.Compared rate of cure,mortality and complications in two groups.Results In treatment group cured 30 cases ( 85.7% ),5 patients died (14.3% ),the control group cured 23 cases (65.7%),death 12 cases(34.3%).Two groups had significant difference (x2 = 4.15,x2 = 3.99,P ＜ 0.05 ).The GOS score (3.23 ±2.15) points in treatment group were lower than the control group(5.03 ±0.96) points after treatment(t =3.52,P ＜0.05).The incidence of complications in the treatment group was 17.0% (6/35) lower than the control group,37.1% (13/35 ) (x2= 3.95,P＜0.05).Conclusion Subhypothermia care may reduce complications in patients with severe brain injury.

Aim In order to observe the pathological features and the dynamic distribution of RH strain T. gondii in main organs of infected mice, using indirect immunoenzymatic technique. To provide pathological diagonsising reference of toxoplasmosis and increase to understand the pathologensis of Toxoplasmosis. Methods Mice were infected intraperitoneally with 103 tachyzoites of T. gondii RH strain,and the parasites were detected using indirect immunoenzymatic technique in the liver, spleen, lungs and brain at 2,4,6 and 8 days postinfection. Results The liver was the first organ parasitized at D2, followed by spleen and lungs at D4, the brain at D8. At the early phase of infection, parasites were found on the edge of the liver and spleen. A few parasites were detected within the liver, spleen and lungs with time being. But parasites increased progressively and distributed well during the whole phase. The brain was the last organ to be parasitized. Parasites multiplicated rapidly so that the mice were seriously ill and died. Conclusions The indirect immunoenzymatic technique can demonstrate tachyzoites and Toaxoplasma antigen clearly in infected mice during acute stage. Many organs were infected such as liver, spleen,lungs and brain. The results suggest that the organs in the peritoneal cavity were infected directly by tachyzoites as IP infection, then the parasites disseminate through a blood way, and in the end, tachyzoites cross the blood-brain barries to reach the brain.

ObjectiveTo observe the clinical effect of warm needling based on conventional rehabilitation in treating cerebral palsy.MethodSixty patients with cerebral palsy were randomized into an observation group and a control group, 30 in each group. The control group was intervened by physiotherapy (PT), occupational therapy (OT), speech therapy (ST), conductive education (CE), etc.; the observation group was by mild moxibustion at Baihui (GV20), Zusanli (ST36), and Guanyuan (CV4) in addition tothe above rehabilitation therapies.ResultThe rehabilitation result in the observation group was superior to that inthe control group (P<0.05).ConclusionMild moxibustion can significantly enhance the rehabilitation result in patients with cerebral palsy.

Objective: To establish an LC-MS/MS method for the determination of 4-( 4-amino-3-fluorophenoxy )-N-methylpyri-dine-2-carboxamide ( AFP-PMA) as a genotoxic impurity in regorafenib. Methods: The content of AFP-PMA was determined by an LC-MS/MS method. A Waters XBridge Shield RP18 column was adopted to separate the samples and the column temperature was 50℃. The mobile phase consisted of 5 mmol·L-1ammonium acetate aqueous (A)-acetonitrile (B) with gradient elution (0~9 min, 5%B→90%B) at a flow rate of 1. 0 ml·min-1. An electrospray ionization source (ESI) was used in a positive-ion and multiple reactions monitoring mode. The ion channel was m/z 262. 2→244. 1. Results:The standard curve was linear within the range of 2. 41-980. 90 ng·ml-1(r=0. 9998) and the limit of quantification was 8. 02 ng·ml-1. The limit of detection was 2. 41 ng·ml-1, which was e-quivalent to 0.000241% for the concentration of regorafenib. The average recovery was 100.95% and RSD was 2.37% (n=9). Conclusion:The method has good specificity, promising accuracy and high sensitivity, which can be used for determining the trace genotoxic impurity AFP-PMA in regorafenib.