Objective To explore the influence of Akt inhibitor MK2206 in the proliferation and apoptosis of tongue squamous cell carcinoma TCA-8113 cells,and to clarify the possible mechanism.Methods The tongue squamous carcinoma TCA-8113 cells at the logarithmic phase were randomly divided into control group and 1,5,25,125, 250 nmol·L-1 MK2206 groups.The inhibitory rate of proliferation of TCA-8113 cells was detected with MTT method,and the apoptotic rate of TCA-8113 cells was determined with flow cytometry(FCM),and the expressions of caspase-9,Bad,GSK-3β,p-Akt and T-Akt proteins in the TCA-8113 cells were detected with Western blotting method.Results The IC50 of tongue squamous cell carcinoma TCA-8113 cells after treated with MK2206 for 12, 24,and 36 h were (112.54±1.67),(79.67±2.01),and (33.33±1.98)nmol·L-1 .The FCM results showed that the apoptotic rates of TCA-8113 cells after treated with 1,5,25,125,and 250 nmol·L-1 MK2206 for 12 h were (14.2±0.74)%,(19.3±0.45)%,(35.1±0.45)%,(39.6±0.48)% and (52.1±0.19)%;there were significant differences compared with control group(P<0.01).The Western blotting method results showed that the expressions of p-Akt, Bad and GSK-3βwere decreased with the increasing of dose and time of MK2206;compared with theβ-actin in control group,the bands got darken;the expression level of caspase-9 was increased, compared with theβ-actin in control group, the bands got darken;the T-Akt protein expression did not change significantly;compared with the β-actin in control group, the color of bands had no significant difference.Conclusion Akt inhibitor MK2206 can inhibit the proliferation of tongue squamous cell carcinoma TCA-8113 cells and induce apoptosis.

Objective To analyze the effect of conductive education combined with massage on activity of children with severe cerebral palsy. Methods 11 children (7 males and 4 females) with cerebral palsy receiving rehabilitation therapies in CereCare Wellness Centre of Xuhui, Shanghai from April to October 2010 were enrolled in this study. The type of cerebral palsy was as follows: 7 tetraplegia, 1 diplegia,2 athetoid, and 1 dystonia. The average age was 7.47 years, ranging from 4.7～11.9 years. The participants were divided randomly into 2groups. 5 children in group A accepted conductive education and massage therapy in the first 3 months, and then accepted routine rehabilitation therapies in the next 3 months. 6 children in group B accepted routine rehabilitation therapies in the first 3 months, and then accepted conductive education and massage therapy in the next 3 months. The phase of conductive education combined with massage was called intervention period, and the phase of routine rehabilitation therapies was called contrast period. The gross motor function was assessed with Gross Motor Function Measure Scale (GMFM), while the fine motor function was assessed with Fine Motor Function Measure Scale (FMFM).Pediatric Evaluation of Disability Inventory (PEDI) was used to assess activities of daily living. The different effects between conductive education combined with massage and routine rehabilitation therapies were compared. Results The scores of GMFM and FMFM both increased in intervention period, while both decreased in contrast period. The score of FMFM increased significantly (P<0.05). The scores of PEDI changed without statistical significance. Conclusion Conductive education combined with massage may increase the fine motor function of children with severe cerebral palsy compared with routine rehabilitation therapies.

Objective To study the male reproductive ability of male rats with Toxoplasma gondii ( Tg) infection and investigate the variation of Toxoplasma infection in seminal plasma of infertile patients and explore its mechanism. Methods Thirty SD rats were randomly divided into 3 groups. The rats in the Toxoplasma infection group were administrated intraperitoneally with tachyzoites of Tg. in a dosage of 2 × 10~ 5/ml(2ml) , the rats in the treated group were administered with the same dosage of the tachyzoites and from the second day after the infection they were treated with 200 mg/kg azithromycin for 7 days, and the normal group was given physiological saline. Nine weeks after the infection, the serum sex hormone level, number,vitality, activity and quantity of spermatozoa and activities of enzymes in testa's of the testicular tissues were determined in the male rats. The female rats infected with Tg were matched with normal female rats at a ratio of 1: 2 for one week, and on the 21st day of pregnancy, the number of corpora luteum, sex ratio and the weight, body length and tail length of fetus were measured. The ELISA method was used todetermine the seminal plasma's anti-Tg IgG antibody of the 169 patients with infertility and 35 males with normal fertility. Meanwhile the NO levels in their semina were determined by means of nitric acid reducase. Results The number, activity .vitality, serum level of sex hormones were all lower in the infected rats than those in the normal and treated groups. The number of fetus in the pregnant rats matched with the infected male rats was significantly fewer, but the average body weight, body length, tail length of the fetuses and sex proportion showed no significant difference in comparison with those of the control group. The anti-Toxoplasma gondii antibody positive rate in the masculine infertility patients was 18.35% , being significantly higher than 2. 86% in the normal fertility group(P < 0.05 ). The mean NO level in the semina from the infertility group was (146.68 ± 38. 87) μnol/L , which was significantly higher than (84.92 ± 26.72) μnol/L( P < 0.01) in the fertility group. Conclusion Toxoplasma gondii infection can cause certain influences on the male reproductive ability.

Objective To examine the expressions of IKKε protein in the specimens and cells of epithelial ovarian cancer and investigate the effect of IKKε inhibitor on cell proliferation and apoptosis. Methods (1) A total of 118 cases of patients with the median age of 59 who have accepted surgical treatment due to ovarian cancer in the First Affiliated Hospital of Dalian Medical University from January 2006 to April 2013 were selected. Twenty cases of patients with the median age of 55 who have accepted hysterectomy and salpingo-oophorectomy due to uterine leiomyoma during the same period were selected as the control. The expressions of IKKε protein were detected by immunohistochemistry in normal ovarian tissues and epithelial ovarian cancer specimens,and the relationship between the expressions of IKKε and the clinical features of patients was analyzed. IKKε protein was determined by western blot in various ovarian cancer cells, including SKOV3, OV2008, C13, A2780S, A2780CP, OV4, OV5, OV8, and CAOV3 treated with or without IKKε inhibitor. The cellular proliferation and apoptosis of ovarian cancer cells after 48 hours treatment of IKKε inhibitor were analyzed by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry, respectively. Results (1) The immunohistochemical results showed that IKKε was highly expressed in epithelial ovarian cancer specimens with the expression rate 66.1% (78/118), compared with normal ovarian tissue with the expression rate 35.0% (7/20), which exhibited statistically significant difference (χ2=6.993, P=0.008). The expression of IKKε protein was correlated with International Federation of Gynecology and Obstetrics (FIGO) stage, histological grade, the level of CA125 in preoperative serum and distribution of the tumor (P<0.05), but no correlation with age, histological type, the incidence pattern, and tumor size (all P>0.05). (2) IKKε was widely overexpressed in different levels in SKOV3, OV2008, C13, A2780S, A2780CP, OV4, OV5, OV8, and CAOV3 cells, and the expression of IKKε decreased as the increase of the concentration of IKKε inhibitor (0.1 and 0.5 μmol/L) in OV2008, C13, A2780S, and A2780CP cells after 48 hours treatment. Different concentrations of IKKε inhibitor (0.05, 0.1, 0.5, 1, 5, 10, and 25 μmol/L) significantly inhibited the proliferation of OV2008, C13, A2780S, A2780CP, and SKOV3 cells in a concentration-dependent manner (P<0.05), and the half maximal inhibitory concentration (IC50) was 0.43, 0.86, 0.10, 0.19, and 0.24 μmol/L, respectively. The cell apoptotic rate of OV2008, C13, A2780S, A2780CP, and SKOV3 cells was significantly increased after 48 hours treatment of IKKεinhibitor with the concentration of 0.1 and 0.5 μmol/L (P<0.05). Conclusions The IKKε protein in epithelial ovarian cancer specimens and cells is overexpressed. IKKε inhibitor could inhibit cellular proliferation and induce apoptosis in a concentration-dependent manner. Together, the result indicated that IKKε may be a candidate target for the treatment of ovarian cancer in future.

Objective To investigate the possible correlation between the monocyte chemoattractant protein-1 ( MCP-1 ) gene A-2518G single nucleotide polymorphism (SNPs) in the promoter region and acute coronary syndrome (ACS) in Chinese Han ethnic population of Sunan region,Methods This study was conducted with a case-control design in 484 ACS patients including 290 acute myocardial infarction (AMI)patients and 194 patients with unstable angina pectoris (UAP) and 346 control subjects ruled out coronary disease by coronary angiography (control group),including 166 patients with coronary atherosclerosis and 180 subjects without coronary stenosis.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used for the detection of the A-2518G polymorphism in MCP-1 gene,and then thefrequency of genetype was statistically analyzed.Results There were AA,AG and GG genotypes of MCP-1 gene A-2518G polymorphism in the ACS group and control group.The two groups could be considered as a genetic equilibrium representative by Hardy-Weinberg equilibrium ( P ＞ 0.05 ).Compared with the control group,the frequencies of AA genotype ( 15.32％ vs.16.12％ ),AG genotype (53.47％ vs.51.86％ ),GG genotype (31.21％ vs.32.02％ ) and G allele genotype (57.95％ vs.57.95％ ) in ACS group were not significantly different ( P was 0.083,0.673,0.821 and 1.00,respectively).Multivariate logistic regression analysis indicated that there was no significant correlation between MCP-1 gene A-2518G polymorphism and ACS regardless of differences in gender,age,smoking,diabetes,TG and LDL-C ( P ＞0.05 ).There was no significant difference in gender and age of ACS onset between two groups ( P ＞ 0.05).There were no significant differences in the frequencies of AA,AG and GG genotypes and G allele genotype among AMI group,UAP group and normal coronary group ( P ＞ 0.05).Conclusions The data shows that MCP-1 gene A-2518G polymorphism is not associated with the risk of ACS in the Chinese Han ethnic population living in Sunan region.

This article investigated the applicability of the near infrared spectroscopy (NIR) technology, combined with the least squares support vector machines (LS SVM) used for the quality monitoring of medicated leaven fermentation. First, near infrared spectra of 67 medicated leaven samples were obtained by near infrared spectroscopy system in the wavelength range of 400 2 500 nm, and then the protease and amylase activity were measured by Folin phenol method and DNS method. Thereafter, the LS SVM was employed to calibrate models. The Rc and Rp of protease in near infrared model were 0.975 and 0.938, respectively; The RMSEC and RMSEP were 5.297 and 9.795, respectively. The Rc and Rp of amylase in near infrared model were 0.987 and 0.973, respectively; The RMSEC and RMSEP were 7.215 and 6.864, respectively. This model has good prediction ability and is suitable for quality monitoring in medicated leaven fermentation process. The research achievement could lay a certain foundation for the near infrared spectral analysis technology applied in the field of traditional fermentation processing.

This study was aimed to optimize bio-solid bidirectional fermentation conditions of Trametes robiniophia Murr. for rhubarb. Sulphuric acid-phenol colorimetry, magnesium-methanol colorimetry, and HPLC were used in the
content determination of polysaccharide, total anthraquinone, and 4 free anthraquinones. The drying rate and con-sumption rate were combined as indicators for the optimization of technical parameters such as medicinal dosage, temperature and amount of water. The results showed that when using 500 mL conical flask, the best fermentation conditions were medicinal dosage of 10 g, the temperature of 34℃, adding water of 120%. It was concluded that bidirectional fermentation of rhubarb increased the content of free anthraquinones. Among them, the content of chrysophanol with anti-oxidation effect increased significantly. The decreasing of combined anthraquinone can relieve the severe laxative effect of rhubarb.

Objective To investigate the mechanism of fluoroquinolone resistance in clinical isolates of Enterococcus faecium. Methods The MICs of six fluoroquinolones(norfloxacin,ciprofloxacin,ofloxacin,levofloxacin,gatifloxacin and moxifloxacin) against 35 clinical isolates of E.faecium from eight hospitals in Tianjin were determined by agar dilution method in the absence or presence of multidrug resistance efflux pump inhibitor reserpine.The quinolone-resistance determining region(QRDR)of parC and gyrA were amplified and sequenced.Results No less than twofold decrease in MIC values of the six fluoroquinolones in the presence of reserpine was observed in 35,29,1,0,6 and 2 of the 35 strains of E.faecium respectively.One fluoro- quinolone-susceptible isolate and five fluoroquinolone-resistant isolates were selected randomly to analyze the QRDR of parC and gyrA.All five fluoroquinolone-resistant isolates had single amino acid alteration in both GyrA and ParC.Ser-80 in ParC was substituted by lie(4 isolates)or Arg(1 isolates).Glu-87 in GyrA was replaced by Lys(2 isolates)or Gly(2 isolates). The other one had an Ser-83-to-Ile substitution.The one fluoroquinolone-suseeptible isolate had no alteration in the QRDR of either ParC or GyrA.Conclusions Both target alteration and active efflux are responsible for the resistance to fluoroquinolone in clinical isolates of E.faecium.

Objective To explore the anatomical characteristic of the third palmar interosseous mus- cle as well its dominate nerve,and to investigate the anatomical basis of difficult recovery of digitus minimus adduction.Methods Twenty aduh fresh hands without deformity and trauma were obtained.Dissect and observe the third palmar interosseous muscle and its dominate nerve and adjacent structure under surgical mi- croseope,measure the size of the third pahnar interosseous muscle and its dominate nerve,the data were pro- cessed by stastistics method.Results Among palmar interosseous muscles and its dominate nerves,the third palmar interosseous muscle and its dominate nerve is the smallest.There are conspicuous tendon bundle on the surface of the third palmar interosseous muscle partly,which have a potential compression on the third palmar interosseous muscle dominting nerve.Conclusion The third palmar interosseous muscle is the smal- lest among palmar interusseous muscles and it is the only digitus minimus adduction muscle.The sominating nerve of the third palmar interosseous muscle is small anti the tendon bundle of the third palmar interosseous muscle have a potential compression.All these can cast light on diffcult recovery of digitus minimus adduction.

Objective To investigate the influence of anti-angiogenesis therapy on proliferation and apoptosis of fibroblasts derived from keloids. Methods Thirty pieces of keloids from a patient were implanted into subcutaneous tissue of the nude mice, 24 pieces of which survived were divided into three groups which were treated with perilesional injection of vascular endothelial growth factor( VEGF) (0.4 mg/0.2 mL) , Endostar(0.125 g/0.2 mL) and physiological saline (0.2 mL)on the 21 d, 23 d, 25 d, 27 d after implantation. Sample were collected on the 10th day after perilesional injection, the proliferating fibroblasts in keloid tissue were immunohistochemically detected by proliferating cell nuclear antigen (PCNA) expression. The apoptotic cell was detected by terminal deoxynucleotidyl transferase dUTP-nick end labeling (TUNEL) staining. Results IHC staining indicated that PCNA expression of fibroblasts was significantly increased in keloid tissue after VEGF injection, PCNA expression of fibroblasts was significantly reduced in keloid tissue after Endostar injection,TUNEL assay revealed lower apoptotic cells expression in the keloid tissue after VEGF injection and higher in the Endostar group than control group. The rate of proliferative index (PI) , apoptotic index(AI) and AI/PI of fibroblasts in keloid after VEGF (PI:41.13 ±2.29,AI:5.75 ±1.28,AI/PI: 0.14 ± 0.04)or Endostar injection (PI:27.25 ±2.61,AI:11.00±1.31,AI/PI:0.41 ±0.09)and control group (PI: 34.75 ±3.62,AI:7. 88 ± 1.64,AI/PI:0. 23 ±0.07) showed statistical differences. Conclusion Anti-angiogenesis therapy is shown to induce keloid regression through suppression of keloid fibroblast proliferation,induction of apoptosis, which may be a new approach for the treatment of keloids.