Objective To investigate the effect of tourniquet compression on axonal transport in sciatic nerve of rats.Methods Twenty-four 12-week old male SD rats weighing 250-300 g were randomly divided into 4groups according to the duration of tourniquet compression(n=6 each):1,2,4 and 12 h.The tourniquet was applied to the middle 1/3 of thigh.In each animal whether the left or right thigh was compressed was determined by a flip of coin.The tourniquet was released for 10 min after every hour of compression.A 3-cm segment of sciatic nerve was removed at the end of tourniquet compression(1.5 cm proximal and 1.5 cm distal to the site of compression).Immuno-histochemistry was used to measure the expression of insulin-like growth factor-1(IGF-1)in the sciatic nerve.The ratio of average optic density of the compressed sciatic nerve to that of control was used to estimate the degree of IGF-1 accumulation.The regression equation of the interaction between the duration of compression and accumulation of IGF-1 was analyzed.Results There was significant accumulation of IGF-1 in the sciatic nerve proximal to the compressed site.The accumulation increased with the duration of compression.There was no significant accumulation of IGF-1 in the sciatic nerve distal to the compressed site.The regression equation of the interaction between the duration of compression(X)and accumulation of IGF-1(Y)was Y=0.422X+0.887.Conclusion Tourniquet compression of sciatic nerve can inhibit axonal transport.The accumulation increases with the duration of compression.

Objective To investigate the effects of brain ischemic preconditioning (BIP) on peripheral blood EPCs and neovascularization in ischemic brain tissue in rats with cerebral ischemia reperfusion injury (IRI). Methods One hundred and eight male SD rats were randomly divided into three groups:SO group (n=36), MCAO group (n=36) and BIP group (n=36). Neurological function assessment was conducted at 0 h before MCAO-reperfusion, 3 h, 24 h and 3 d, 5 d as well as 7 d after MCAO-reperfusion (n=6 for each group in each time point). Flow cytometry was used to calculate the number of EPCs. Immunohistochemical staining was used to detect the capillary density. Results ①Although neurologi?cal deficit scores were significantly decreased in both BIP and MCAO groups after 3 h following MCAO-reperfusion, the scores were much lower in BIP group than in MCAO group(5 d:1.00±0.63;7 d:1.00±0.63, P<0.05).②The numbers of EPCs were decreased in MCAO group while was increased in BIP group at 3 h after MCAO-reperfusion. The numbers of EPCs were significantly higher in BIP group than in MCAO group(24 h:0.58±0.07;3 d:0.80±0.10;5 d:0.68±0.05;7 d:0.52 ± 0.03, P<0.01). ③ The new blood vessels could be detected at 3 d in BIP group and 5 d in MCAO group after MCAO-reperfusion. The numbers of new blood vessels were significantly higher in BIP group than MCAO group(5 d:14.53 ± 3.44; 7 d: 41.40 ± 5.62, P<0.01). ④ Pearson analysis showed a positive correlation between EPCs and capillary density (5 d: r=0.855, P<0.01; 7 d: r=0.946, P<0.01). Conclusion BIP can improve EPCs mobilization and function, which may contribute to neovascularization in the ischemic brain tissue.

Objective To investigate the role of protein phosphorylation in Pseudomonas aeruginosa (P.aeruginosa) strains in response to stress triggered by mouse macrophages.Methods The strong cation exchange-immobilized metal affinity chromatography (SCX-IMAC) was performed to enrich phosphopeptides.The nanoscale liquid chromatography coupled to tandem mass spectrometry (nano LC-MS/MS) was carried out to identify and analyze phosphoproteome.Results Fourteen phosphopeptides from twelve proteins were identified within thirty-one phosphorylation sites on serine,threonine and tyrosine residues.Fifty percent of these phosphorylated proteins were membrane proteins,indicating that their phosphorylation modification was more critical for bacteria in response to the stress.In terms of biological process of Gene Ontology,these identified proteins were involved in stress response,iron transport,anaerobic respiration,response to hydrogen peroxide and signal transduction by phosphorylation,etc.Conclusion These phosphorylated proteins in P.aeruginosa strains are necessary for signal transduction and their response to harsh environment within the macrophages,such as iron limitation,hypoxia and oxidative stress.This study provides evidence for further investigation on virulence and pathogenesis of P.aeruginosa.

Objective To investigate the differential MSCT diagnostic features and comparative study of subtypes of renal cell carcinoma (RCC). Methods All of the renal cell carcinomas including 14 chromophobe RCCs (ChRCC), 10 papillary RCCs type 1(PRCC Ⅰ), 15 papillary RCCs type 2 (PRCC Ⅱ), 7 mucinous tubular and spindle cell carcinomas (MTSCCs) were investigated except for clear cell RCC. Dynamic contrast-enhanced CT was conducted in each case after intravenous administration of contrast agent, and the data including all the CT manifestations and the enhancement features were analyzed and contrasted together. Results The indexes including enhancement homogeneity, border of the tumor, renal pelvis violation, blood vessel in tumor showed statistically significant difference between the 4 subtypes (P<0.05), but no difference in the calcification of the tumor. Only the enhancement degree of MTSCC was lower than the kidney medulla in all of the three enhancement scanning phases, while the other 3 tumors' enhancement degree was higher than the kidney medulla in the cortical phase. Peak contrast enhancement of ChRCC was located in the cortical phase, however, peak contrast enhancement of the others did in the nephrographic phase. Conclusions Enhancement characteristics combined CT features is of great help in differential diagnosis of 4 subtypes of RCC.

Objective Lysine-specific demethylase 1 (LSD1) is a flavin adenine dinucleotide-dependent oxidase, which participates in many biological processes , such as cell proliferation and differentiation and gene activation and repression .The aim of this study was to investigate LSD1 acetylation by histone deacetylase inhib -itor trichostatin A ( TSA) and its effect on TSA-induced apoptosis of ovarian cancer cells . Methods LSD1 shRNA was synthesized and implanted into the pLKO-Tet-On lentiviral vector , which was transfected into HO8910 and SKOV3 ovarian cancer cell lines , and then the transfected cells were screened with 1.5μg/mL puromycin for one week until stable clones were established .The cells were treated for 48 hours with methanol (2 mg/mL, control), TSA (200 nmol/L), TCP (100μmol/L), or TSA+TCP.And in the experiment of RNA interfering the LSD1 expression, the cells were also treated for 48 hours with methanol (2 mg/mL, control), TSA (200 nmol/L), Dox (100 ng/mL), or TSA +Dox.The levels of LSD1 acetylation and its substrate histone H3 lysine 4 dimethylation (H3K4me2) were de-tected by immunoprecipitation (IP) and Western blot.The apoptosis of the cells was determined by Annexin Ⅴ/PI staining and flow cytometry, the transcription levels of the Bax and p21 genes detected by real-time quantitative PCR, and the H3K4me2levels in the promoter regions of Bax and p21 measured by chromatin immunoprecipitation ( ChIP ) .Results In comparison with the methanol control, the TSA group showed significantly increased levels of LSD 1 acetylation in the HO8910(1.00±0.29 vs 5.83±0.46, P<0.01) and SKOV3 cells ( 1.00±0.24 vs 5.07±0.35, P<0.01) as well as that of H3K4me2 ( P<0.01);the total apoptosis rates of HO 8910 and SKOV3 cells were remarkably increased in the TSA, TCP, and TSA+TCP groups (P<0.05), even more significantly in the TSA+TCP than in the TSA and TCP groups ( P<0.05) .The mRNA expressions of Bax and p21 in the HO8910 cells were markedly upregulated in the TSA, Dox, and TSA+Dox groups (P<0.05), even more significantly in the latter than in the former two groups (P<0.05).The TSA group exhibited a higher level of H 3K4me2 than the methanol control in the promoters of Bax(2 .92±0.26 vs 0.68±0.19, P<0.01) and p21 (3.07±0.29 vs 0.93±0.17, P<0.01). Conclusion TSA induces the LSD1 acetylation, while suppression of LSD1 expres-sion and activity may enhance the antitumor activity of TSA .

Background Pathologic myopia is one of the common blinding eye diseases.Recent research suggests that immune response participates in the pathogenesis of pathologic myopia,and inflammation is an important factor that influent immune status.Objective Present study was to observe the change of high sensitive C-reactive protein(hs-CRP) in serum in the patients with pathologic myopia and explore the role of inflammation in the development of pathologic myopia. Methods Serum hs-CRP was measured from 30 patients with pathologic myopia,30 patients with simple myopia and 30 normal controls with Nephelometric Turbidity in the OLYMPUS AU5400 automatic biochemical analyzer.Written informed consent was obmined from each subject before medical examination.Results The mean age was(30+10) years in pathologic myopia group,and(32+8)years in simple myopia group and(32+9)years in normal control group.The range of preoperative spherical equivalent refraction was (-6.00--22.00) D in pathologic myopia group,(-1.00--6.00) D in simple myopia group and(-1.00-+1.00) D in normal control group.The level of hs-CRP in serum was(3.68±1.15)mmol/L in the patients with pathologic myopia and was significantly higher than that of simple myopia group(1.99±0.68 mmol/L)and normal controls (2.11±O.66 mmol/L)(q=10.69,P＜0.01;q=9.91,P＜0.01),respectively.No significant correlation was found between hs-CRP level and myopic degrees in pathologic myopia group(R2=0.037,P＞0.05). Conclusion Hs-CRP may play rule in the inflammatory reaction during the pathogenesis of pathologic myopia.

To establish an EDTA complexation extraction pretreatment combining with GFAAS method for the determination of residual aluminium ion in Huoxiang zhengqi pellets without digestive treatment, systematical investigation was made on sample preparation, and EDTA was used for the complexation extraction of residual aluminium ion in samples. The pH, concentration and volume of extraction solution, the temperature and time of microwave extraction, and graphite furnace temperature program were investigated. The results were compared with the microwave digestion. It was showed that, 0.1 g of sample weight was added in 20 mL 0.05 mol x L(-1) EDTA solution (pH 3.5), followed by heating at 150 degrees C for 10 min in the microwave extraction device. The determination of GFAAS was performed at optimized detection wavelength (257.4 nm) as well as graphite furnace temperature program, the detection limits and quantification limits were 2.37 μg x L(-1) and 7.89 μg x L(-1), respectively. The precision (RSD) was less than 2.3%. The average recovery was 96.9% -101%. The present method is easy, rapid and accurate for the determination of residual aluminium ion in Huoxiang zhengqi pellets.
Aluminum ; chemistry ; isolation & purification ; Drug Contamination ; Drugs, Chinese Herbal ; chemistry ; Edetic Acid ; chemistry ; Graphite ; chemistry ; Spectrophotometry, Atomic ; methods ; Temperature

Objective To study the clinical manifestations,diagnosis and surgical treatment of tubular adenoma in the extrahepatic bile duct.Methods The clinical data of 3 patients with tubular adenoma in the extrahepatic bile duct who were admitted to the Affiliated Hospital of Inner Mongolia Medical University from July 2010 to February 2012 were retrospectively analyzed.Results The tumors were located at the middle and lower part of the common bile duct.All the 3 patients received resection of the common bile duct and Roux-en-Y cholangioenterostomy,and recovered well.The results of pathological examination showed that all the patients were with tubular adenoma,and 2 patients had moderate atypical hyperplasia.Conclusions Tubular adenoma in the extrahepatic bile duct could be diagnosed by pathological examination.Radical resection and rerouting of the bile duct can get a better outcome.

Objective To explore the toxicity and biological activity of cobalt nanoparticles (CoNPs) on osteoclasts,and to analyze the relationship between cobalt nanoparticles and osteolysis.Methods From November 2014 to July 2015,RAW264.7 cell was induced to osteoclast-like cell by LPS.Different concentrations of cobalt nanoparticles and cobalt chloride were added,and the cell morphology was observed under a microscope.24 h after induction on RAW264.7,cells were grouped into cobalt nanoparticles group (10,20,50,100 μmol/L),cobalt chloride group (10,20,50,100 mol/L) and control group.MTT assessment and Q-PCR were performed at 2 h,4 h,8 h,24 h,48 h post-treatment.Results With the increase of concentration (10,20,50,100 μmol/L) and the action time (2 h,4 h,8 h,24 h,48 h),the inhibition rate of cobalt nanoparticles and cobalt chloride on osteoclast like cells was significantly increased,and the inhibition rate of cobalt nanoparticles was higher.With different concentrations (10,20,50,100 μmol/L) of CoNPs and cobalt chloride,the relative expression of CAⅡ,Cat K gene mRNA expression decreased compared with the control group,when the concentration of CoNPs was in the range of 10-50 μ mol/L,the relative expression of CAⅡ and Cat K was increased,which was reduced in cobalt chloride group.Conclusion Different concentrations of cobalt nanoparticles and cobalt chloride can inhibit the proliferation and differentiation of osteoclasts,and cobalt nanoparticles is more pronounced,when the concentration of cobalt nanoparticles was 10-50 μmol/L,the relative expression of osteoclasts CAⅡ,Cat K increaseed,which was suppressed at the same concentration of cobalt chloride.

Objective Protein arginine methyltransferase 5 (PRMT5),a member of the histone arginine methylation transferase family,is involved in a wide range of biological regulation through ei-ther epigenetic or posttranslational methylation modifications. The pur-pose of the present study was to investigate the effects of PRMT5 on cell proliferation of ovarian cancer cell HO8910. Methods Cell lines HO8910 with PRMT5 overexpression were obtained by transi-ent transfection,which were divided into three groups in the experiment: blank control group (wild-type cell line HO8910),negative control group (HO8910 cells were transfected with pCMV-myc plasmid),and experimental group (HO8910 cells were transfected with pCMV-myc-PRMT5 plasmid). Western blot was used to detect the expression of myc protein,and qRT-PCR was used to detect the ex-pression of PRMT5 mRNA. Cell lines HO8910 with inducible stable knockdown of PRMT5 were established by shRNA interference method,which were divided into four groups: pLKO control group (infected by empty vector lentivirus),pLKO+Dox (100ng/mL) group,shPRMT5 group (infected by PRMT5shRNA lentivirus) and shPRMT5+Dox (100 ng/mL) group. Western blot and qRT-PCR were used to detect the expression of PRMT5 protein and mRNA levels. Dox-induced PRMT5 knockdown was detected by increasing Dox concentration,which includes four groups,Dox 0ng/mL group,Dox 1ng/mL group,Dox 10ng/mL group,Dox 100ng/mL group,and each group was treated for 48 hours. Western blot and qRT-PCR were used to detect the PRMT5 protein and mRNA expression. Colony formation assay,EdU assay,and CCK-8 assay were used to test cells proliferation. The experiment was conducted in two large groups each with two subgroups: PRMT5 knockdown group (Dox-group,Dox+ group),PRMT5 overexpression group (pCMV-myc group,pCMV-myc-PRMT5 group). Western blot was used to detect the effects of PRMT5 expression on proliferation-related proteins. The experiment was conducted in two large groups,PRMT5 knockdown group with four subgroups : Dox 0ng/mL group,Dox 1ng/mL group,Dox 10ng/mL group and Dox 100ng/mL group,and PRMT5 overexpression group with two subgroups (pCMV-myc group and pCMV-myc-PRMT5 group). Results Western blot results showed that the expression of myc was detected in the experimental group in which HO8910 cells were transfected with pCMV-myc-PRMT5,and the expression of PRMT5 mRNA was significantly higher in the experimental group than those in the blank control group and the negative control group (P<0.001) . Western blot and qRT-PCR showed that PRMT5 protein (0.32±0.25) and mRNA expression levels in shPRMT5+Dox group were significantly lower than those of shPRMT5 group (0.89±0.18) (P<0.05). Western blot and qRT-PCR confirmed that PRMT5 protein (0.21±0.24) and mRNA expres-sion in Dox 10ng/mL group and Dox 100ng/mL group (0.08±0.15) were significantly downregulated compared to Dox 0ng/mL group (1.11±0.15) (P<0.05). Colony formation experiments,EdU experiments,and CCK-8 experiments confirmed that the proliferative ca-pacity of cells in Dox+group was lower than that of Dox-group in PRMT5 knockdown group(P<0.05); while in PRMT5 overexpression group,the proliferative capacity of pCMV-myc-PRMT5 group was significantly higher than that of the pCMV-myc group (P<0.05). Western blot results showed that the protein expression of Cyclin D1 was significantly lower in Dox 100 ng/mL group (0.17±0.06) than that in Dox 0 ng/mL group (1.18±0.18) (P<0.05) and the expression of P21 was significantly increased in PRMT5 knockdown group (P<0.05). In the PRMT5 overexpression group,the protein expression of Cyclin D1 in pCMV-myc-PRMT5 group (3.48± 0.22) was higher than that in pCMV-myc group (0.88±0.15) (P<0.05),while the protein expression of P21 (0.08±0.17) were significantly lower than that of pCMV-myc group (4.12±0.10) (P<0.05). Conclusion PRMT5 plays an important role in the proliferation of ovarian cancer cells. Down-regulation of PRMT5 can inhibit cell proliferation and up-regulation of PRMT5 can pro-mote cell proliferation.