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

Inflammation plays an essential role in the pathophysiology of atherosclerosis. Our study was aimed to investigate whether salvianolate, a novel water-soluble phenolic compound of Danshen, alleviates atherosclerosis via regulating the inflammation in rats. High fat diet feeding plus vitamin D3 injection was used to induce atherosclerosis in rats. Salvianolate (60, 120 or 240 mg/kg) or placebo was given to atherosclerotic rats. The plasma lipids, interleukin 6 (IL-6) and C reactive protein (CRP) were measured by ELISA. CD4+CD25+Foxp3+ cells were determined by flow cytometry. Histological changes were examined by hematoxylin and eosin staining. The results showed that the levels of plasma IL-6 and CRP were elevated in the rats fed on high fat diet, and the histological analysis demonstrated the successful establishment of atherosclerosis models. Treatment with salvianolate alleviated the atherosclerotic process and decreased the levels of plasma IL-6 and CRP. Also the number of CD4+CD25+Foxp3+ cells was increased in salvianolate-treated rats. It was concluded that salvianolate could treat atherosclerosis via modulating the inflammation at cytokine and cell levels.

Multidrug resistance proteins (MRP2, ABCC2) may play a role in drug resistance in epilepsy by limiting gastrointestinal absorption and brain access of antiepileptic drugs (AEDs). We sought to investigate the effects of ABCC2 polymorphisms on plasma carbamazepine (CBZ) concentrations and pharmacoresistance in Chinese patients with epilepsy. ABCC2 rs717620, rs2273697, rs3740066 polymorphisms were genotyped by polymerase chain reaction amplification followed by restriction fragment length polymorphism analysis or direct automated DNA sequencing in 80 patients treated with CBZ monotherapy. There were no differences in CBZ maintenance doses or adjusted plasma CBZ concentrations among the ABCC2 rs717620, rs2273697 and rs3740066 genotypic groups. No associations between all the studied genotypes and haplotypes involving the three SNPs of ABCC2 and CBZ resistance were observed in this patient cohort. These results suggest that ABCC2 polymorphisms may not contribute to interindividual variabilities in CBZ daily maintenance doses, plasma concentrations, and treatment efficacy.
Epilepsy

OBJECTIVETo investigate the protective effect and mechanism of curcumin derivatives B06 on myocardium from type 2 diabetic rats.

METHODSThirty-five male SD rats were randomly divided into 5 groups, normal control group (NC group), high fat group (HF group), high fat treatment group (FT group), diabetes mellitus group (DM group) and diabetes treatment group (DT group) (n = 7). The late four groups were fed with high fat food, after four weeks of high fat feeding, the rats from DM group and DT group were injected with low dosage of streptozocin intraperitoneally to induce diabetes mellitus, FT group and DT group were gavaged with curcumin derivatives B06 at the dosage of 0.2 mg/kg x d. The blood glucose and lipid were detected biochemically, blood insulin was assayed by ELISA and the insulin resistance index was calculated, the morphology of myocardium was observed by light and transmission electron microscopy, the protein expression of AMP-activated protein kinase alpha (AMPKalpha) and phosphorylated AMP-activated protein kinase alpha (p-AMPKalpha) in myocardium were tested by Western blot.

RESULTSThe level of blood glucose, lipid, insulin and the insulin resistance index were increased in HF group and DM group, but they were decreased after the treatment with B06. The expression of AMPKalpha and p-AMPKalpha were decreased, but they became increased after the treatment of B06. There were increased collagen fibers in interstitium and expansion of mitochondria in cytoplasm of myocardium from DM group, but they were ameliorated in B06 treatment group.

CONCLUSIONIt is suggested that B06 may relieve the damage of myocardium from type 2 diabetic rats and the increased expression of AMPKalpha and p-AMPKalpha may be involved in it.

AMP-Activated Protein Kinases ; metabolism ; Animals ; Blood Glucose ; Curcumin ; pharmacology ; Diabetes Mellitus, Experimental ; physiopathology ; Heart ; drug effects ; Insulin Resistance ; Male ; Myocardium ; pathology ; Rats ; Rats, Sprague-Dawley ; Streptozocin

Inflammation plays an essential role in the pathophysiology of atherosclerosis. Our study was aimed to investigate whether salvianolate, a novel water-soluble phenolic compound of Danshen, alleviates atherosclerosis via regulating the inflammation in rats. High fat diet feeding plus vitamin D3 injection was used to induce atherosclerosis in rats. Salvianolate (60, 120 or 240 mg/kg) or placebo was given to atherosclerotic rats. The plasma lipids, interleukin 6 (IL-6) and C reactive protein (CRP) were measured by ELISA. CD4+CD25+Foxp3+ cells were determined by flow cytometry. Histological changes were examined by hematoxylin and eosin staining. The results showed that the levels of plasma IL-6 and CRP were elevated in the rats fed on high fat diet, and the histological analysis demonstrated the successful establishment of atherosclerosis models. Treatment with salvianolate alleviated the atherosclerotic process and decreased the levels of plasma IL-6 and CRP. Also the number of CD4+CD25+Foxp3+ cells was increased in salvianolate-treated rats. It was concluded that salvianolate could treat atherosclerosis via modulating the inflammation at cytokine and cell levels.
Animals ; Atherosclerosis ; blood ; etiology ; prevention & control ; C-Reactive Protein ; metabolism ; Cholecalciferol ; administration & dosage ; Diet, High-Fat ; adverse effects ; Dose-Response Relationship, Drug ; Flow Cytometry ; Forkhead Transcription Factors ; metabolism ; Inflammation ; blood ; prevention & control ; Interleukin-6 ; blood ; Lipids ; blood ; Lymphocyte Count ; Male ; Phytotherapy ; Plant Extracts ; pharmacology ; Rats, Wistar ; Receptors, Complement 3b ; metabolism ; Salvia miltiorrhiza ; chemistry ; T-Lymphocytes, Regulatory ; drug effects ; metabolism ; Vitamins ; administration & dosage

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

Perioperative stroke is cerebral infarction occurring in the perioperative period.The incidence of perioperative stroke in non-cardiac,and non-neurologic surgery is about 0.7％,but the mortality can be as high as 26％ to 40％.The outcome of the patients with perioperative stroke can be disastrous.Here we report a case of perioperative ischemic stroke that occurred after surgery of lumbar decompression and pedical screw fixation.A 76-year-old female admitted to our hospital because of lumbar spinal stenosis.Her medical history included hypertension and diabetes for ten years.Her personal history included a smoking history of 60 years by 2 cigarettes per day,not quitting.Her carotid artery ultrasound showed multiple low echo plaques on the right side and multiple high echo plaques on the left side of the carotid artery,but without distinct stenosis.Other examinations and tests showed no distinct abnormality.She went on a lumbar decompression and pedical screw fixation uneventfully.The blood loss was 400 mL and autologous blood transfusion 150 mL.The arterial blood pressure (ABP) maintained during 100-130 mmHg/60-80 mmHg (1 mmHg =0.133 kPa).Sixty minutes after she recovered from general anesthesia,the patient developed symptoms of slurred speech and right limbs weakness.The anesthesiologist evaluated the patient immediately with National Institute of Health Stroke Scale (NIHSS).The NIHSS score was 11 and a stroke was highly suspected.The acute stroke team was therefore initiated and fast responded.Within 4 h,digital subtraction angiography (DSA) was proceeded,which showed the M1 segment of the left middle cerebral artery was occluded and the local stenosis of her right middle cerebral artery was up to 80％.After the successful embolectomy by Solitaire stent,the left middle cerebral artery reflowed and the forward blood flow was thrombolysis in myocardial infarction (TIMI) grade 3.The patient was discharged after 33 days after the surgery with a NIHSS of 9.Our case provides an example that an acute stroke team that included the department of anesthesiology can be beneficial to the patients' perioperative strokes.During the perioperative period,anesthesiologists should be included into the acute stroke team,because anesthesiologists and anesthesia nurses might be first observers of those early onset strokes.Our case also put forward this thought that a standard peri-operative stroke evaluation tool,like NIHSS,should be discussed and applied to facilitate and accelerate the initiation of perioperative acute stroke team.

Objective To evaluate the accuracy and safety of pedicle screw placement for spinal deformity under intraoperative CT-as-sisted.Methods The clinical data of 28 patients with spinal deformity,who underwent intraoperative CT-assisted placement of pedicle screws in our hospital from March 2014 to March 2017,were analyzed retrospectively.Intraoperative CT-assisted placement of pedicle screws was to observe the position of pedicle screws and adjust the screw position.The safety of pedicle screw fixation was assessed by the postoperative complications.Results All 28 patients of scoliosis were successfully performed surgery.A total of 402 pedicle screws were placed in the spines and the placements of 64 screws of them were adjusted according to intraoperative CT findings.Of 402 screws,377 were placed in the pedicles,13 by grade 1 pedicle breach,8 by grade 2 pedicle breach and 4 by grade 3 pedicle breach according to Gertzbein-Robbins classifi-cation of pedicle screw placement accuracy.No complication was directly related to the screws placement.No patient required reoperation due to the misplacement of the pedicle screws.Conclusion Intraoperative cone-beam CT scan can help the surgeon improve the accuracy and safety of pedicle screw placement for spinal deformity with less complication.

BACKGROUNDThe incidence of prostate carcinoma (Pca) has been increasing in China. We detected Pca in elderly men in Changchun, north China and the significance of prostate specific antigen (PSA) in mass screening and clinical staging of Pca.

METHODSSerum PSA from 12,027 men over 50 years old from Changchun was analyzed. In case of serum PSA greater than 4.0 ng/ml, the patient was suspected of potentially suffering from Pca, and transrectal six-point puncture prostate biopsies were performed under ultrasound guidance. Pathological examinations were performed on the biopsy tissue, and ABCD and TNM clinical stagings were used in accordance with international standards. Correlations between serum PSA level and clinical stage were analyzed.

RESULTSPSA was greater than 4.0 ng/ml in 813 patients (6.8% of the 12,027 men). Transrectal six-point prostate puncture biopsies guided by ultrasound were performed in 273 patients (33.6% of the 813 patients who were tested positive in the initial mass screening). Of these 273 patients, 69 cases of Pca (25.3% of 273) were confirmed by biopsy in the second screening, with an overall detection rate for Pca of 0.57% (69/12,027). The total number of patients in stages A, B, T1, or T2 was 57.9%, and over 20% of them suffered from late stage Pca with lymph node and bone metastasis. An obvious positive correlation was observed between ABCD staging, TNM staging, and serum PSA level.

CONCLUSIONSSerum PSA level is not only the golden standard for mass screening of Pca, but also the predictor for clinical stage of Pca. PSA testing revealed asymptomatic Pca cases in early, middle, and later stages in the elderly, suggesting that mass screening is of paramount importance.

Aged ; Aged, 80 and over ; Biopsy, Needle ; Humans ; Male ; Mass Screening ; methods ; Middle Aged ; Neoplasm Staging ; Prostate-Specific Antigen ; blood ; Prostatic Neoplasms ; diagnosis

OBJECTIVETo observe the treating effect of collage-heparin sulfate after the 10 mm rat sciatic nerve defect was bridged by it.

METHODSA new kind of nervous tissue engineering scaffold was produced by freeze-drying technique from collagen-heparin sulfate. Thirty-two SD rats were randomly divided into A, B, C and D groups. Sciatic nerve defect in group A was bridged by collagen-heparin sulfate. In group B, sciatic nerve was bridged by auto-nerve transplantation. Group C was the blank control group. Animals in group D were normal. And 10 mm sciatic nerve defect was bridged in the experiment. Thirty-six weeks after the operation, the experimental animals were detected by HRP labeled retrograde trace, HE staining, toluidine staining, silvering staining, S100, GAP-43 and NF immunohistological staining, MBP immunofluorescence staining and transmission electron microscope to observe the nerve regeneration inducing effect of this new scaffold.

RESULTSNine months after operation, the collage-heparin sulfate scaffold was replaced by newly regenerated nerve. The number of HRP labeled spinal cord anterior horn cells and the area of sensation nerve fiber at the posterior horn were similar with that was repaired by auto-nerve. GAP-43, NF and S100 labeled regenerated nerve fiber had passed the total scaffold and entered the distal terminal. The regenerated nerve fibers were paralleled, lineage arranged, coincide with the prearranged regenerating "channel" in the collagen-heparin sulfate scaffold. MBP immunofluorescence staining also proved that the newly regenerated nerve fiber could be ensheathed. In the experimental group, the area of myelinated nerve fiber and the thickness of the myelin sheath had no obvious difference with that of the group repaired by auto-nerve, except that the density of the regenerated myelinated sheath fiber was lower than that of the control group.

CONCLUSIONNervous tissue engineering scaffold produced by collagen-heparin sulfate can guide the regeneration of nerve fibers. The nerve function recovers fine. This kind of material has great application potential.

Animals ; Biocompatible Materials ; Heparitin Sulfate ; Male ; Prosthesis Implantation ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; injuries ; pathology ; surgery ; Sulfuric Acid Esters ; Tissue Engineering ; methods