Objective To study the effect of methotrexate on the neointimal formation after arterial balloon injury. Methods Male rabbits were randomized into four groups: MTX 0.5 mg/kg per week, MTX 1.5 mg/kg per week, MTX 5 mg/kg per week and the control. Drugs were administered by intramuscular injection. Rabbit carotid arteries were harvested 2 and 4 weeks after injury. Results Histologically, the intimal areas were reduced significantly in MTX treated animals compared with the controls. The VSMC proliferation in injured vessels was identified by immunostaining for proliferating cell nuclear antigen (PCNA). In comparison, PCNA-positive cells in both intima and media were significantly reduced by treatment of MTX. But MTX did not enhance reendothelialization in the injured carotid arteries as determined by Evans blue stain.Conclusion Low dosage of MTX could attenuate neointimal formation after arterial injury by inhibiting VSMC proliferation.

A growing number of children and adolescents are being diagnosed as Chiari malformation type Ⅰ (CM-Ⅰ) for behavioral disorders,developmental delay,seizures,or abnormal orpharyngeal function.The aim of this study was to compare the clinical characteristics,imaging findings and surgical outcomes of CM-Ⅰ in pediatric and adult patients.Between January 2014 and June 2017,84 patients with CM-Ⅰ underwent surgical treatment in our department.We divided the patients into two groups:pediatric group (n=1 1,age ＜18 years)and adult group (n=73,age ≥18 years).Data on clinical characteristics,imaging findings,surgical outcomes,and prognosis were retrospectively reviewed and compared between these two groups.For clinical presentation,scoliosis (36.4％) and developmental delay (36.4％) were more common in pediatric patients,whereas,sensory disturbance (58.9％) and motor weakness (41.1％) were more common in adult patients.Imaging findings showed that the incidence of hydrocephalus and craniovertebral junctional abnormalities was significantly higher in pediatric group than in adult group (P＜0.05).Compared to adult group,pediatric group showed a better improvement or resolution of syrinx and tonsillar herniation after surgical treatments (P＜0.05).The total Chicago Chiari Outcome Scale (CCOS) score in pediatric patients at the last follow-up was significantly higher than that in adult patients (P=0.002).In conclusion,the clinical characteristics and imaging findings appeared to be different in pediatric and adult patients with CM-Ⅰ.The surgical outcomes of pediatric patients were shown to be significantly better than those of adult patients.

Aim To observe the effect of methotrexate (MTX) on proliferation, migration and apoptosis of cultured vascular smooth muscle cells (VSMC). Methods Rabbit thoracaortic VSMC were cultured in vitro.VSMC proliferation was evaluated by cell counting and cell cycle analysis. Monolayer cell scrape was used to observe VSMC migration. Apoptosis was observed with flow cytometry, DNA gel electrophoresis and TUNEL stain. Results MTX (25～100 nmol?L -1) inhibited VSMC proliferation in a dose-dependent manner.25 nmol?L -1 and 50 nmol?L -1 MTX increased the percentage of the S phase cells and decreased the percentage of the G 2/M phase cells (P

OBJECTIVETo assess the characteristics of enhanced magnetic resonance image with ultrasmall superparamagnetic iron oxide (USPIO) in the inflammatory and tumor metastatic rabbit model, and explore its relevance with histologic ultrastructural findings.

METHODSTotally 36 New Zealand white rabbits were randomly divided into lymphadenitis group and metastatic group. Complete Freund's adjuvant was injected into the bilateral dorsal footpads of 18 rabbits to set up ipsilateral lymphadenitis model. The other 18 rabbits received a subcutaneous implantation of VX2 tumor cell suspension (1.5 x 10(7) cells/ml) in both thighs to set up metastatic lymph node model. Magnetic resonance scan were performed 24 hours before and after USPIO (90 micromol Fe/kg) injection. T2 values of each lymph node were measured and lymph node T2 enhancement rate was calculated as well. HE staining, Prussian blue staining, and electronic microscopy were performed to observe the pathological microstructure changes and the distribution of the iron particle in lymph node. Relationship between lymph nodes USPIO enhancement and its microstructures were further analyzed. Results Thirty-six lymph nodes in lymphadenitis group showed different degrees of reactive hyperplasia. Twenty-six lymph nodes in metastatic group were invaded by tumor cell. Non-enhanced scan showed mild difference between T2 signal intensity of the two pathological lymph node types. After USPIO enhancement, inflammatory lymph nodes showed distinct T2 signal reduction at the center, and metastatic lymph nodes showed homogenous and faint T2 signal reduction. Enhancement rate of benign and malignant lymph nodes were 57.39% and 29.45% respectively (P < 0.01). HE staining and Prussian blue staining indicated USPIO particles located mainly in the macrophages at inflammatory lymphatic medulla, while paracortical area and cortical area contained relatively much less USPIO particles due to less macrophages distribution. MRI findings were correlated with the pathological results. Electronic microscopy also verified that the majority of USPIO particles were located in the numerous cytophagic bubbles of macrophages. Lymph nodes metastasis including 4 lymph nodes with completed structure destruction due to entire tumor infiltration, 19 lymph nodes with partially lymph node structure destruction but reduced USPIO-contained macrophage numbers or reduced USPIO particles in macrophages, and 3 lymph nodes with only localized foci tumor metastasis at subcapsular area. Conclusions USPIO enhancement pattern of different lymph nodes is closely related to distribution and functional status of the intra-node macrophages. It may affect the accuracy of the lymph node property diagnosis based on USPIO enhanced image.

Animals ; Dextrans ; metabolism ; Female ; Image Enhancement ; methods ; Lymph Nodes ; ultrastructure ; Lymphadenitis ; diagnosis ; pathology ; Lymphatic Metastasis ; diagnosis ; ultrastructure ; Magnetic Resonance Imaging ; methods ; Magnetics ; Magnetite Nanoparticles ; Male ; Nanoparticles ; Rabbits ; Random Allocation

Background:Mutations in the isocitrate dehydrogenase 1 (IDH1) and IDH2 genes are important for both the integrated diagnosis and the prognosis of diffuse gliomas.The p.R132H mutation of IDH1 is the most frequently observed IDH mutation,while IDH2 mutations were relatively rarely studied.The aim of the study was to determine the pathological and genetic characteristics of lowergrade gliomas that carry IDH2 mutations.Methods:Data from 238 adult patients with lower-grade gliomas were retrospectively analyzed.The status of IDH1/2 gene mutations,telomerase reverse transcriptase (TERT) promoter mutations,O6-methylguanine-DNA-methyltransferase (MGMT) promoter methylation,1p/19q co-deletion and the expressions of IDH1 R132H,alpha-thalassemia X-linked mental retardation,and p53 were evaluated.Progression-free survival (PFS) and overall survival (OS) were calculated via Kaplan-Meier estimation using the log-rank test.Results:Totally,71％ (169/238) of patients were positive for IDH mutations,including 12 patients harboring mutations in IDH2.Among the 12 patients with IDH2 mutations,ten patients harbored the R172K mutation,one patient harbored the R172S mutation and one harbored the R172W mutation.Of these,11 tumors occurred in the frontal lobe and showed morphology typical of oligodendroglioma.The proportion of grade Ⅱ tumors was higher than that of grade Ⅲ tumors in IDH2 mutant-gliomas.IDH2 mutations were frequently associated with TERT promoter mutations,1p/19q co-deletion and MGMT promoter methylation.IDH2 mutations were associated with better outcomes compared with IDH wild-type gliomas (P ＜ 0.05).However,the PFS and OS did not differ from that of IDH1 mutant patients (P =0.95 and P =0.60,respectively).Conclusions:IDH2 mutations are more frequent in oligodendrogliomas and associated with a better prognosis.IDH2 mutations may segregate in distinct clinico-pathological and genetic subtypes of gliomas,and therefore may merit routine investigation.

AIMTo investigate the effects of morphine on synaptic transmission of neurons of central nervous system and reveal the mechanism underlying it.

METHODSNew born wistar rats were used for primary culture of hippocampus neurons. Using whole-cell patch-clamp technique, we observed the excitatory and spontaneous inhibitory postsynaptic current (EPSC, sIPSC) and glutamate-induced current before and after morphine treatment.

RESULTS(1) sEPSC of hippocampal neurons was markedly increased after morphine application. The effect of morphine was blocked by opioid antagonist naloxone (n=18, P < 0.01). (2) The frequency of mEPSC and the amplitude of glutamate-induced current of hippocampal neurons had no significant changes after morphine treatment (P > 0.05). (3) Morphine inhibited sIPSC of hippocampal neurons markedly and naloxone could block this effect (n=13, P < 0.01).

CONCLUSIONThe results suggest that the exciting effect of morphine on hippocampal neurons are not due to direct influence of morphine on glutamate synapses transmission, but may result from the inhibition on interneurons, that is "disinhibition" way.

Animals ; Animals, Newborn ; Cells, Cultured ; Excitatory Postsynaptic Potentials ; physiology ; Hippocampus ; cytology ; Inhibitory Postsynaptic Potentials ; Morphine ; pharmacology ; Neurons ; drug effects ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Wistar ; Synaptic Transmission ; drug effects ; physiology

OBJECTIVETo study the effect of THW-4, an extract from Trypterygium hypoglaucum on proliferation and apoptosis of vascular smooth muscle cells (VSMC).

METHODSVSMC derived from rabbit aorta were cultured in vitro and different concentrations of THW-4 were added in experimental groups. Cell proliferation was evaluated by MTT and apoptosis by transmission electron microscopy (TEM), TUNEL assay and Annexin V-FITC/PI double labelled assay.

RESULTSThe inhibitory effects of THW-4 on proliferation of VSMC displayed dose-time dependently, with the IC50 value of 15.6 microg/L at 48 hrs. Incubated with THW-4 (10-100 microg/L) for 56 hrs, VSMC mainly appeared early stage apoptosis and the percentage of apoptosis was found to raise along with the increase of the THW-4 concentration. Typical images of apoptosis could be observed under TEM and TUNEL assay showed increase of DNA segments with karyorrhexis and pyknosis after THW-4 treatment for 72 hrs. Analysis of cell cycle indicated the THW-4 mainly lead to the blockage of VSMC in G2/M stage.

CONCLUSIONTHW-4 could inhibit the proliferation and induce the apoptosis of VSMC in vitro, suggesting that THW-4 is a potential agent for prevention of restenosis following angioplasty.

Animals ; Aorta ; cytology ; Apoptosis ; drug effects ; Cell Division ; drug effects ; Cells, Cultured ; Coronary Restenosis ; prevention & control ; Drugs, Chinese Herbal ; pharmacology ; Male ; Muscle, Smooth, Vascular ; cytology ; Rabbits ; Tripterygium ; chemistry

BACKGROUNDUric acid (UA) is suspected to play a neuro-protective role in Parkinson's disease (PD). This study aimed to evaluate whether the serum UA level was associated with the disease progression of PD in a relatively large population of Chinese patients.

METHODSSerum UA levels were measured from 411 Chinese PD patients and 396 age-matched controls; following the uric acid colorimetric method, the serum creatinine (Scr) levels were also measured to reduce the bias caused by possible differences in renal excretion function. The disease progression was scored by Hoehn and Yahr (H&Y) scales and disease durations; PD group was divided into 3 subgroups according to H&Y scales. Independent-samples t test was performed to analyze the differences between PD group and control group. Multiple analysis of covariance was performed to analyze the differences between PD subgroups. Spearman rank-correlation was performed to evaluate the associations between serum UA or Scr level and disease progression.

RESULTSPD patients were found to have significantly lower levels of serum UA than controls ((243.38 ± 78.91) vs. (282.97 ± 90.80) µmol/L, P < 0.01). As the disease progression, the serum UA levels were gradually reduced. There was a significantly inverse correlation of UA levels with H&Y scales (Rs = -0.429, P < 0.01) and disease duration (Rs = -0.284, P < 0.01) in PD patients of both females and males. No significant difference of the Scr level between PD patients and controls was found ((70.01 ± 14.70) vs. (69.84 ± 16.46) µmol/L), and the Scr level was not involved in disease progression.

CONCLUSIONLower serum UA levels may possess a higher risk of PD, which may be a potential useful biomarker to indicate the progression of PD.

Aged ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Parkinson Disease ; blood ; pathology ; Uric Acid ; blood

Objective To investigate the pharmacokinetic and bio-equivalence of domestic and imported recombinant human insulin injec-tion in Beagle dogs.Methods Twelve Beagle dogs were assigned into two groups and the two groups were separately singlely given with test preparation and reference preparation in the same dose in sc injection method according to a randomized two -phase crossover .The plasmas were sampled at different time points after subcutaneous administration and the blood glucose levels were determined by Roche Glucose meter synchronously.The radioimmunoassay ( RIA) method was used to deter-mine the concentration of blood insulin at the different sample points . The pharmacokinetic parameters were calculated by DAS 2.0 software. Results The main pharmacokinetics parameters of test preparation and reference preparation were as follows: t1/2 were ( 1.06 ±0.17 ) and (1.06 ±0.35 ) h; Cmax were ( 97.10 ±45.20 ) and ( 91.30 ±28.20 )μU· mL-1; tmax were ( 0.49 ±0.19 ) and ( 0.49 ±0.18 ) h; area AUC0-t were (168.00 ±40.50) and (168.00 ±41.90) μU· mL-1 · h, respectively. What′s more, Cmin of the test and the referenc were (1.36 ±0.28 ) and (1.37 ±0.34 ) mmol· L-1 separately, and the tmin were ( 1.57 ±0.55 ) and ( 1.74 ±0.65 ) h, respectively. The confidence intervals of Cmax and AUC0-t are 85.5% -117.7% and 97.0%-103.3%.Conclusion The results showed that the test and the reference preparation meet the regulatory cri-teria for the pharmacokinetic equivalence .

Adult ; CD5 Antigens ; metabolism ; Carcinoma, Papillary ; metabolism ; pathology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Choristoma ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Hamartoma ; metabolism ; pathology ; Humans ; Proto-Oncogene Proteins c-kit ; metabolism ; Thymoma ; metabolism ; pathology ; Thymus Gland ; pathology ; Thymus Neoplasms ; metabolism ; pathology ; Thyroid Neoplasms ; metabolism ; pathology ; surgery ; Thyroidectomy ; methods