Objective To compare the condition of illness and pathological characteristics of experimental autoimmune encephalomyelitis (EAE)in C57 BL/6 mouse models induced by myelin oligodendrocyte glycoprotein 35-55 (MOG35-55)at different doses,and provide a reliable animal model for further study of multiple sclerosis(MS).Methods Male SPF-grade C57 BL/6 mice were divided randomly into four groups:normal group and three EAE model groups (MOG35-55 high-dose,middle-dose and low-dose model groups).200,100,50μg MOG35-55/mice were mixed with complete Freund's adjuvant(CFA),respectively,to prepare complete antigen in different concentrations.The mice were anesthetized and injected s.c.over flanks with the complete antigen and injected i.P.with pertussis toxin to establish immunization-induced C57BL/6 mouse-model of EAE.The mice of the normal group were injected with normal saline instead.Since the day of immunization,the incidence,body weight and neurological score of the mice were observed.The mice of different neurological scores in different periods were anesthetized and perfused with saline and followed by 4% paraformaldehyde.The brain and spinal cord of the mice were removed and fixed in the same fixative solution.The brains and spinal cords of the mice were examined by histopathology with hematoxylin-eosin(HE) staining.The mice on the 40th day were sacrificed and perfused with 2% paraformaldehyde and 2% glutaraldehyde, 1 mm~3 pieces of cerebral white matter and intumescentia lumbalis of the spinal cord were taken and ultrathin sections were prepared according to conventional techniques for electron microscopy. Results All the MOG_(35-55) in three different doses induced mouse models of EAE. The disease was with an incidence rate of 100% and a chronic monophasic course. The body weight of the mice in the three groups decreased obviously compared with those in the normal group. The maximum value of neurological score was 1.33,2.25 and 2.50 in the mice of high-, middle-and low-dose groups, respectively. The major histopathological changes observed in the brain and spinal cord of the EAE mice were different degrees of inflammatory cell infiltration around small vessels showing sleeve-like changes, dcmyelination and neuronal karyopyknosis in the acute and remission stages. The main site of the brain inflammation was in white matter around encephalocoele, and also in the DG and CA zones of hippocampus. The spinal cord inflammation was most severe in the lumbosacral region. The above mentioned pathological changes in the low-dose group were more prominent than those in the middle-dose and high-dose groups. The major ultrastructural changes were scattered around encephalocoele, interstitial edema, especially around small blood vessels, and swollen mitochondria with damaged cristae, and some karyopyknosis in vascular endothelial cells. Some tight junctions were blurred. Some dispersed lymphocytes and mononuclear cells were seen in the perivascular space. In lumbar intumescentia of the spinal cord, there were some myelin figures in the white matter myelin sheath. Some of them showed demyelization and structurtal fusion. The cytoplasmic organelles of axons were considerably reduced or even disappeared. The vascular basement membrane showed an increased thickness and focal necrosis in some areas. Conclusion The mouse models of immune-induced EAE are successfully established with MOG_(35-55), especially that induced with MOG in a dose of 50 μg. This mouse model is stable, with a high incidence and low mortality rate, and can be applied for EAE research in the future.

ObjectiveTo investigate the prevalence and distribution characteristics of dental fluorosis of children aged 8 - 12 in Wushan and Fengjie counties of Chongqing and to provide a scientific basis for prevention and control of the disease.MethodsTwenty townships(towns) in Fengjie county and 18 in Wushan county were selected as survey points by random cluster sampling in 2010.Dental fluorosis of all the children aged 8 - 12 was examined with Dean index.The detection rate of children's dental fluorosis,defect rate and dental fluorosis index were compared between the two counties.Results Totally 38 209 children aged 8 - 12 were investigated.The total detection rate of dental fluorosis was 43.09％( 16 466/38 209) in the two counties.The detection rates of dental fluorosis in Wushan and Fengjie were 48.98％ (9397/19 186)and 37.16％(7069/19 023),respectively,and the difference was statistically significant(x2 =544.03,P ＜ 0.01 ).Total detection rates of dental fluorosis of the five age groups(8,9,10,11,and 12-year-old age groups) were 32.52％(2157/6632),40.07％(2672/6668),43.67％(3420/7831 ),46.01％ (3861/8391) and 50.14％ (4356/8687),respectively,and the difference was statistically significant (x2 =510.50,P ＜ 0.01),Dental fluorosis indexes in Wushan and Fengjie were 0.713 and 0.485,respectively.Defect rates of dental fluorosis in Wushan and Fengjie were 4.05％ (777/19 186) and 1.57％(298/19 023),respectively.Conclusions The total detection rate of dental fluorosis of the two counties is still high,which gradually increases with age.Wushan is still an endemic area of dental fluorosis,and Fengjie is at the edge of the

Objective To investigate the diagnostic significance of endoscopic findings in Henoch-Schonlein purpura(HSP),especially when abdominal pain preceded the cutaneous lesions.Methods The clinical data and gastroscopic findings in 37 cases of children with HSP were studied and analysed retrospectively in order to detect the pathological changes in the stomach and duodenum mucosa.The biopsy was taken in the pathological changeing place,and the relationship between clinical and endoscopic findings was analyzed.Results Detection rate of the pathological changes in the stomach and duodenum mucosa was 62.2%,31.3% of which experienced only cutaneous lesions,100% of which presented the acute abdominal pain.Three patients were not checked up the pathological changes.Of them,1 had arthritis,2 had Henoch-Schonlein nephritis.Characteristically endoscopic findings in the stomach and duodenum mucosa were found.The endoscopic findings included anabrosis,hyperemia,edema and hemorrhage.Conclusions Detection rate of the pathological changes in the stomach and duodenum mucosa is higher.Endoscopy is very helpful to the early diagnosis of HSP in children,especially abdominal pain presented firstly.

Objective To evaluate the application of plasma drug monitoring in pediatric HIV/AIDS patient antiretroviral therapy adherence monitoring.Methods Totally 261 plasma samples and related information were collected from three consecutive follow-up visits of 87 HIV-infected children treated in Shangcai county CDC of Henan province from March to October 2009.The plasma concentrations of antiretroviral drugs were measured by a developed high performance liquid chromatography-mass spectrometry method.Potential adherence influencing factors, such as regimen, age, gender, parent conditions, previous ART exposure and therapy duration, were analyzed by univariate logistic regression.Results Plasma concentration of antiretroviral drugs lower than LLTR (1 000 ng/ml) was the criteria to identify missed dose.The concentrations of 28 plasma samples were lower than LLTR, which meant missing dose.There were 17 patients (19.5%) with their concentrations lower than LLTR at least once in three follow-up visits.Logistic regression analysis of adherence related factors showed that compared with the children whose parents were both alive, the children whose mother and (or) father died were more likely to miss dose.The odds ratio was 4.13(95% credibility interval:1.37-12.46, P values was 0.012).Conclusions HIV-infected children have adherence problems when receiving antiretroviral therapy.Plasma therapeutic drug monitoring can be one of the effective methods to monitor the adherence.

Objective Based on middle cerebral artery occlusion (MCAO) model,to investigate the effects of melatonin (MT) on DWI and expression of Fas,FasL and cleaved Caspase-3 proteins in rat model with focal cerebral ischemia.Methods Eighty SD rats were randomly divided into Sham group (n=16),MCAO group (n=32) and MT group (n=32).The rats in sham group were treated with sham-operation.And the rats in MCAO and MT groups were peritoneally injected with saline and MT respectively.The behavioral scores were assessed in the three groups.The rats in MCAO and MT group with the behavioral scores of 1 3 points were selected in the study.The DWI relative signal intensity (rDWI-SI),Fas,FasL and cleaved Caspase-3 proteins were respectively examined by MR scaning and immunohistochemical staining in all rats of each group at 6 h,24 h,72 h and 7 days after ischemia reperfusion (IR) or sham-operation.And the DWI and immunohistochemical results for each group were compared.Results At last,there were 16 rats in sham group,29 rats in MCAO group and 30 rats in MT group,respectively.There was significant difference of the behavioral scores among the three groups (x2 =50.125,P＜0.01).The behavioral scores of MT and MCAO groups were higher than those of sham group (all P ＜0.05).And the behavior scores of the MT group were lower compared with MCAO group after IR.Compared with the rDWI-SI values measured at 6 h,24 h and 72 h,7 days in sham group,the rDWLSI values of MT and MCAO groups were significantly higher (all P＜0.01).And the rDWI-SI was higher in MCAO group than those in MT group at 6 h,24 h and 72 h after IR (all P＜0.01).And there was no significant difference of rDWI-SI at 7 days after IR between MT and MCAO groups (P＞0.05).The immunohistochemical staining results showed that the number of Fas,FasL and cleaved Caspase-3 positive cells in MCAO and MT groups were significantly higher than those in sham group (all P＜0.01).And there were less Fas,FasL and cleaved Caspase-3 positive cells in MT groups compared with MCAO group (all P＜0.05) at 6 h,24 h and 72 h after IR.There was no significant difference of Fas,FasL and cleaved Caspase-3 positive cells among the three groups at 7 days after IR (P＞0.05).Conclusion MT can effectively alleviate the rDWI-SI value and inhibit the expression of Fas,FasL and cleaved Caspase-3 proteins in rats of focal cerebral ischemia.

OBJECTIVE:To establish a method for the plasma concentration determination of carboplatin,and study the phar-macokinetics of carboplatin in female rats after intravenous injection and intraperitoneal injection. METHODS:HPLC was per-formed on the column of Agilent TC-C18 with mobile phase of methanol-water(5:95,V/V)at a flow rate of 1.0 mL/min,detection wavelength was 229 nm,and column temperature was 25 ℃. The inner standard was 5-bromouracil,and injection volume was 20 μL. 24 SD rats were randomly divided into 4 groups,6 in each group. The rats were intravenously injected and intraperitoneally in-jected carboplatin 20,40 mg/kg respectively. 0.5 mL blood sample was taken from eyes before administration and after administra-tion of 0.25,0.5,1,1.5,2,4,6,8,10,12 h. The plasma concentration of carboplatin was determined,and DAS 2.0 was used to calculate the pharmacokinetic parameters. RESULTS:The linear range of carboplatin in plasma was 0.30-60.00 μg/mL (r=0.9991);RSDs of intra-day,inter-day precision were lower than 10%(n=5);RSD of peak area in stability test was lower than 10%(n=5);method recovery was 98.7%-102.4%(RSD≤6.08%,n=5),and extraction recovery was 83.38%-85.45%(RSD≤5.97%,n=5). AUC0-12 h of carboplatin 20,40 mg/kg by intravenous injection and intraperitoneal injection in female rats were (15.503 ± 4.172),(23.402 ± 4.266),(6.716 ± 2.306),(9.384 ± 2.205)μg·h/mL;AUC0-∞ were (16.424 ± 4.846),(23.404 ± 4.266),(6.790±2.378),(9.765±2.095)μg·h/mL;t1/2z were(1.246±0.765),(0.394±0.058),(0.513±0.156),(0.884±0.460) h;and tmax were(0.700±0.274),(0.400±0.335),(0.542±0.368),(0.833±0.289)h,respectively. CONCLUSIONS:The meth-od is simple,economic and accurate,with suitable internal standard,and can be used for the plasma concentration determination of carboplatin in female rats and the pharmacokinetic studies.

ObjectiveTo investigate the differences of whole blood chemical elements and urinary fluorine between patients with endemic fluomsis and patients without endemic fluorosis,and to find out the elements associated with endemic fluorosis and further lay a theoretical basis for clarify the pathogenesis of the disease.MethodsUsing case-control study,100 children aged 8 - 12 with dental fluorosis in Wushan and Fengjie counties of Chongqing from December 2010 to February 2011,and 30 adults with skeletal fluorosis were enrolled as case group; 100 children aged 8 - 12 without dental fluorosis and 30 adults without skeletal fluorosis were enrolled as internal control group; and 50 children without dental fluorosis and 30 healthy adults were selected as external control group in non-epidemic areas in Yubei district.Whole blood copper,zinc,calcium,magnesium,iron and urinary fluorine of all subjects were determined,and differences of these indexes were compared between groups.ResultsThe levels of copper,zinc,calcium,magnesium,iron and urinary fluorine of children in the case group were (30.08 ± 2.83),(74.04 ± 9.75)μmol/L,(1.65 ± 0.29),(1.37 ± 0.17),(6.79 ± 1.27)mmol/L,and (0.73 ±0.37)mg/L,respectively; the levels of these elements of children in internal control group were (28.65 ± 3.96),(72.83 ± 11.35)μmol/L,(1.62 ± 0.27),(1.36 ± 0.18),(6.73 ± 1.22)mmol/L,and (0.48 ± 0.21)mg/L,respectively; in external control group were (32.03 ± 2.99),(77.78 ± 10.85)μmol/L,(1.41 ± 0.11),(1.43 ± 0.13),(7.66 ±0.55)mmol/L,and (0.49 ± 0.26)mg/L,respectively(all P＜ 0.05),the comparison between any two groups indicated the levels of copper,zinc,magnesium,iron of the case group were lower than that of external control group,urinary fluorine was higher than that of internal and external control groups(all P ＜ 0.05).The levels of copper,zinc,calcium,magnesium,iron and urinary fluorine of adult case were (26.93 ± 4.37),(95.89 ± 12.45)μmol/L,(1.50 ± 1.76),(1.56 ± 1.96),(8.15 ± 1.00)mmol/L,and (2.17 ± 0.99)mg/L; internal control group were (26.26 ±4.96),(94.86 ± 12.18)μmol/L,(1.57 ± 0.12),(1.46 ± 0.16),(7.64 ± 1.00)mmol/L,and (1.44 ± 1.22)mg/L;external control group were (26.20 ± 2.96),(96.52 ± 11.11)μmol/L,(1.48 ± 0.14),(1.45 ± 0.16),(7.81 ±0.91 )mmol/L,and (0.55 ± 0.21 )mg/L,respectively.The levels of magnesium,iron and urinary fluorine of case group were higher than that of internal control group,magnesium and urinary fluorine were higher than that of external control group(all P ＜ 0.05).ConclusionsIn vivo anti-fluorine elements are deficient in the areas with endemic fluorosis.Other chemical elements,the environment and genetic factors may be related to the pathogenesisof the disease,which needs a further comprehensive analysis.

Objective To investigate feasibility and effect of external fixator plus steel plate in treatment of open tibiofibular fractures combined with tibial defect.Methods The study involved 21 patients with open fractures of tibia and fibula (15 patients with type Gastilo ⅢA,five with type Gastilo ⅢB and one with type Gastilo ⅢC) with concurrent tibial defect of 2-6 cm.External fixator plus fibular steel plate was performed at the first stage,followed by iliac bone grafting for bone defect at the second stage.Results All patients showed successful reconstruction of the tibial defects with length difference between affected and healthy extremities less than 2 cm in follow-up for (14.0 ± 10.5) months (range,8-24 months).Meanwhile,no talipes equinovarus existed.Conclusions External fixator plus steel plate is an effective method for treating open tibiofibular fractures combined with tibial defect.The length and function of the extremities of patients with open tibiofibular fractures combined with tibial defect of less than 6 cm can be successfully restored.

Objective To observe the effect of magnesium valproate sustained release tablets on children with epilepsy and its effects on cognitive function.Methods Magnesium valproate sustained release tablets were conducted on 38 cases.Close attention was paid to both the degree of paroxysm control and side effects during treatment while periodic examinations on liver function and blood routine were also conducted.The intelligence and P300 of children with epileptics were respectively measured before and after 6-month treatment.Forty children of control group was set up.Results Eighteen cases were totally under controlled(47.4%),11 obviously effect(28.9%),6 effect(15.8%).The total effective rate in total was 92.1%.Obvious differencees in intelligence between children with epileptics and control group before and after 6-month treatment were observed(all P0.05).Conclusions Magnesium valproate sustained release tablets is a new type of broad-spectrum anti-epileptic drug,which has an obvious effect on treatment of children with epileptic without any obvious adverse reaction.It imposes little influence on children′s cognitive function.

Objective To investigate the expression of homeobox gene MSX-2 during cranial suture fusion of SD rats and discuss its significance. Methods SD rats aged 1, 2, 5, 8, 12, 15, 18, 22, 30 and 45 days were selected, and immunohistochemistry and Real-time PCR were employed to localize and quantify the expression of MSX-2 in different regions of cranial sutures. Results MSX-2 expressed in calvarial suture tissues including the extreme ends of the osteogenic fronts and the underlying dura mater. The expression of MSX-2 was low in posterior frontal suture (PF) and sagittal suture (SAG) from postnatal day 1 to day 8 before the initiation of suture fusion, while it was higher in PF than in SAG from postnatal day 12 to day 22 after the initiation of PF suture fusion. The expression of MSX-2 significantly declined in PF and was moderately higher than that in SAG from postnatal day 30 to day 45 after the initiation of suture fusion. Conclusion There is different expression of MSX-2 in PF and SAG during different suture fusion periods, which suggests the expression of MSX-2 may participate in the regulation of cranial bone development and the fusion of cranial sutures.