Objective To investigate the expressions of microRNA-21 in cholangiocarcinoma tissues and the relationship between epithelial-mesenchymal transition (EMT) and the prognosis of patients.Methods Forty-one samples of cholangiocarcinoma and 10 samples of adjacent tissues from 10 patients who received radical resection of cholangiocarcinoma at the Provincial Hospital of Anhui Medical University from January 2005 to January 2010 were collected.The expressions of microRNA-21,E-cadherin and N-cadherin were detected by in situ hybridization and immunohistochemistry,and effect of their expressions on the prognosis was analyzed.Enumeration data were analyzed using chi-square test.The correlation between microRNA-21 and EMT markers was analyzed using the Spearman correlation coefficient.The survival curve was drawn by Kaplan-Meier method,and the survival rate was analyzed using the Log-rank test.Results The expression rate of microRNA-21 in the cholangiocarcinoma tissues was 63％,which was significantly higher than 30％ of that in the adjacent tissues (x2 =0.324,P ＜ 0.05).The expression of microRNA-21 was closely related with the tumor differentiation degree,lymph node metastasis,perineural invasion (x2 =6.365,0.552,11.896,P ＜ 0.05),but not with gender,age,tunor location and tumor type (x2 =0.322,0.588,0.510,0.256,P ＞ 0.05).The expressions of E-cadherin and N-cadherin were related with lymph node metastasis and perineural invasion (x2 =4.630,5.512;6.600,7.152,P ＜0.05),but not with gender,age,tumor location,tumor differentiation degree and tumor type (x2 =0.266,0.013,0.067,0.666,0.003; 1.036,0.997,1.808,2.997,0.812,P ＞0.05).A positive correlation between the expression of microRNA-21 and EMT related markers E-cadherin and N-cadherin was detected (r =0.373,0.614,P ＜0.05).The results of survival analysis showed that the overall survival rate and tumor-free survival rate of patients with low expression of microRNA-21 were significantly higher than those of high expression of microRNA-21 (x2 =3.999,4.376,P ＜ 0.05).Conclusion Over expression of microRNA-21 in cholangiocarcinoma and metastatic lymph nodes may accelerate the invasion and metastasis of cholangiocarcinoma through inducing EMT,microRNA-21 might predict the prognosis of patients.

Objective To investigate the effects of plaque staining test on periodontal health of middle-aged and elderly patients with dental implants. Methods Ninety patients with dental implants were included in the study and divided into test group (n=45) and control group (n=45) by using random digit table. In the test group, the patients were all instructed with both routine health education and dental plaque dyeing education, while the control group received routine health education only. The plaque index (PLI) and gingival index (GI) were assessed at months 1 and 6 after implant restoration. Results One month after dental implantation, there were no significant differences in PLI and GI between the two groups (P>0.05), while 6 months after the implantation, the differences in PLI and GI between them were significantly different ( P<0 . 01 ) . Conclusion The plaque dyeing technology applied in the oral health education to elderly patients with dental implants can reduce plaque index and gingival index so that their periodontal health is enhanced.

Objective To establish chronic stress model of depression in adolescent rats and to examine the effects of different antidepressant treatment on depression and anxiety-related behaviors.Methods Male Wistar rats were given 21-day chronic mild stress (CMS) during their adolescence (postnatal day 30～50, PND30～50).During stress period, rats were treated with fluoxetine (10 mg/kg i.p.) or imipramine (10 mg/kg i.p.), respectively.After stress, rats were tested for behavioral observation using body weight gain, saccharine test, open field and elevated plus-maze (EPM).Results Compared with control/vehicle (n=10) group, stress/vehicle (n=11) group displayed lower weight gain, saccharine preference index and the number of rearing in open field (P<0.05).Antidepressant fluoxetine, but not imipramine reversed anhedonia and the decrease of the number of rearing induced by stress.In addition, compared with early adolescent(PND29) rats, late adolescent (PND52) rats in control/vehicle group exhibited less open arm entries and open arm time, more closed arm time in EPM (P<0.05).Rats in stress/vehicle group showed more open arm entries and less closed arm time than controls(P<0.05).Both fluoxetine and imipramine had no effects on such changes.Conclusions Stress can induce the depression-like behavior in adolescent rats.Fluoxetine, but not imipramine,can effectively reverse anhedonia induced by stress.However, Both antidepressants have no significant effects on stress-induced decrease in developmental increment of anxious behavior during adolescence.These data suggest that chronic mild stress have complicated effects on depressive and anxious behavior in adolescent rats.

To develop an analytic method for qualitative and quantitative analysis of dodecatetraenamides A, B in 42 samples of two official species of Asari Radix et Rhizoma( ARR) (37 samples of Asarum heterotropoides var. mandshuricum with different collection time and 5 samples of Asarum sieboldiivar. seoulense). The HPLC-IT-TOF-MS/MS methods for the qualitative and UPLC-PDA methods for the quantitative analysis were established. Dodecatetraenamides A, B were identified by comparing the retention time, UV absorption spectrum and quasi-molecular ion peak [ M + H]+ with the reference compound using HPLC-IT-TOF-MS/MS. The content of dodecatetraenamides A and B in ARR were determined by UPLC-PDA. The separation was successfully carried out on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 µm) column eluted with mobile phases of water (A) and acetonitrile (B) in gradient program (0-3 min, 35% B; 3-5 min, 35%-36% B; 5-6 min, 36%-43% B; 6 min-11 min 43% B; 11-12 min, 43%-100% B). The column temperature was 45 °C, and the detection wavelength was set at 254 nm. The flow rate was 0.6 mL · min(-1). On one level mass spectrometry scanning, the results showed that the quasi-molecular ion [M + H] + of both dodecatetraenamides A and B were m/z 248.20. The quantitative method with UPLC-PDA has made the baseline separation of the constituents, which were reported as mixtures in the most literatures. The average recovery of dodecatetraenamides A and B were 97.90% and 99.86%, the relative standard deviation were 0.4% and 1.1%, respectively. The contents of dodecatetraenamides A, B in all ARR samples was in the range of 0.11-3.89 and 0.24-6.65 mg · g(-1). Their contents reduced with the extension of storage time. Compared with the samples of 2013, the average content of the two constituents in the samples collected in year 2002-2003 reduced 34% and 36%, respectively (P < 0.05). Compared the A. sieboldii var. seoulense and A. heterotropoides var. mandshuricum with the same collective time and production area, the average contents of the two constituents in latter were up to (1.59 ± 0.75) mg · g(-1) and (2.90 ± 1.17) mg · g(-1), respectively, significantly higher than that in A. sieboldii var. seoulense (dodecatetraenamide A were (0.78 ± 0.52) mg · g(-1), dodecatetraenamide B were (1.69 ± 0.83) mg · g(-1)) (P < 0.05). The content of the dodecatetraenamide A in overground part was in the range of 0.11-0.33 mg · g(-1), dodecatetraenamide B was 0. 24-0.60 mg · g(-1), which were much lower than that of the underground part of ARR (dodecatetraenamide A was in the range of 0.73-3.89 mg · g(-1), dodecatetraenamide B was 2.11-6.24 mg · g(-1)). The method was certified to be simple, accurate and reliable and could be used for qualitative and quantitative analysis of dodecatetraenamide A and B in different species of ARR, also can be used for the comprehensive quality control of traditional Chinese medicine, Asari Radix et Rhizoma.
Amides ; chemistry ; Asarum ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Rhizome ; chemistry

To silence the expression of K-RASAsn12 in human pancreatic cancer cell line by vector-based RNAi(RNA interference) technique,two single-strand DNA sequences encoding mutant-specific shRNA (short haipin RNA) for K-RASAsn12 were synthesized and then inserted into pSilenCircle. The recombinant plasmid was called pSC-K-RASAsn12. According to the same method, pSC-GFP encoding shRNA for GFP was gained. Both recombinant plasmids were transfected into human pacreatic cancer cell line AsPC-1 and BxPC-3. The expression level of K-RASAsn12 was detected by semi-quantitative RT-PCR and Western blot. The result indicated that the recombinant plasmid edcoding mutant-specific shRNA for K-RASAsn12 can inhibit significantly the expression of K-RASAsn12 without affection of wild-type K-RAS(K-RASWT)in Human Pancreatic Cancer Cell Line.

Objective To study the effect of propofol used for outpatient painless enteroscope on cognitive function.Methods One hundred and twenty ASAⅠ~Ⅱpatients scheduled for enteroscope were randomly divided into three groups .Propofol was given 1.5mg/kg(groupⅠ), 2mg /kg (group Ⅱ) or 2.5 mg/kg ( group Ⅲ) intravenously .The enteroscope was inserted when patient showed unconsciousness and no reaction to dictation .SpO2 was kept above 95%96% throughout enteroscope .All patients received neurobehavioral cognitive status examination ( NCSE ) and mini-mental state examination ( MMSE ) test 1 hour before enteroscope examination and 5 minutes,30 minutes, 1 hour after enteroscope examination was o-ver and must finish it within 15 min.The enteroscope examination time , vital signs, analgesia effects and intraoperative awareness were record .Results The ability of memory and calculation at 5 minutes after en-teroscope examination showed a statistical difference between group Ⅰ and ⅡorⅢ( P <0.05),there was no significant difference between in group II and in group Ⅲ( P >0.05 ) , The ability of memory and calcu-lation at 30 minutes, 1 hour after enteroscope examination there was no significant difference in three groups ( P >0.05 ) .In all patients ,the MMSE scores at 5 minutes after enteroscope examination were significant-ly lower than the baseline value ( P <0.05).The MMSE scores at 30 minutes, 1 hour after enteroscope examination in Ⅲgroup patients were significantly lower than the baseline value ( P <0.05 ) .The MMSE scores at 30 minutes, 1 hour after enteroscope examination in I group patients were significantly higher than that inⅡor Ⅲgroup( P <0.05).The MMSE scores at 30 minutes, 1 hour after enteroscope examination there was no significant difference between in group II and in group Ⅲ( P >0.05 ) .The NCSE and MMSE scores at 3hour, 12 hour after enteroscope examination there was no significant difference between in group I and II or Ⅲ( P >0.05).Conclusion Propofol 1.5mg/kg used for painless enteroscope examination has no effect on cognitive function .MMSE and NCSE are suitable for evaluation of outpatient's cognitive func-tion.

Objective To investigate the mechanism of hypoxia regulate osteopontin (OPN) secreting by mature dendritic cells (mDCs). Methods CD14 + cells were enriched using anti-CD14 immunomagnetic beads, for inducing to mDCs, CD14 + cells were cultured with GM-CSF and IL-4 in hypoxia or normoxiain vitro. Concentration of OPN and TGF-β1 in supernatant were detected by sandwich ELISA, OPN mRNA detected by RT-PCR. Approach regulating function of A2 R in expressing of OPN by mDCs by using NECA (surrogate of adenosine), A2R agonist (CGS21680), A2R antagonist (SCH58261) and investigate role of TGF-β1 in this process by using rhTGF-β1 and anti-TGF-β1 Ab. Results Hypoxia inreased the level of OPN and OPN mRNA in mDCs, and this effect could be reversed by A2 R antagonist. Under normoxia,both NECA and A2R agonist (CGS21680) could upregulate the level of OPN and OPN mRNA in mDCs significantly, but this positive effect could be reversed by A2 R antagonist. A2 R played a role in regulating TGF-β1, and confirmed TGF-β1 involved in regulation of OPN by using rhTGF-β1 and anti-TGF-β1 Ab. Conclusion High adenosine induce the generation of TGF-β1 through the A2R on mDCs, and then TGF-β1 raise the OPN secreting by mDCs.

OBJECTIVETo investigate clinical features and the diagnosis by fluorescence in situ hybridization (FISH) of 22q11 microdeletion syndrome (22q11 DS).

METHODSThe clinical data of suspects were analyzed, and their peripheral blood samples were tested by FISH for microdeletion of 22q11. The diagnosis and correlated clinical factors of 22q11 DS were investigated by using the multiple factor Logistic regression analysis and Chi-square test.

RESULTSIn 64 suspects, 14 were shown to have 22q11 microdeletion with many different types of malformation, and the percentage was 21.9%. The Logistic regression predictive equation for 22q11 DS was: y=-8.206+2.324x1+2.725x2+1.674x3, P=exp(y)/[1+exp(y)], in which the concomit ant variables were facial dysmorphic features (x1), congenital heart defects (x2), thymus scarcity/infection problem (x3), the P value meant the probability of diagnosis of 22q11 DS.

CONCLUSIONAccurate clinical evaluation is just as preliminary screening to patients at risk for del22q11. The results of FISH test can be predicted by using the suitable Logistic regression equation.

Adolescent ; Adult ; Child ; Child, Preschool ; Chromosome Deletion ; Chromosomes, Human, Pair 22 ; genetics ; DiGeorge Syndrome ; diagnosis ; genetics ; pathology ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Infant ; Logistic Models ; Male

A new viral sampling concentration device was designed which was equipped with a new cationic filter membrane-Nanoceram suitable for field sampling. Norovirus Genegroup II was detected from environmental water with the aid of this device. The effects on virus recovery of prefiltration, various second-concentration methods, and different eluants were investigated through pre-experiment. The concentration optimized process, and the optimal concentration process were then determined. The results showed that the prefiltration had a profound effect on virus recovery, and two second-concentration method: PEG-NaC1 precipitation and celite adsorption, had almost the same concentration effects. The Na2 HPO4 solution of 0.15 mol/L was selected as the final eluant to elute the adsorbed Nuorovirus from the celite. The virus recovery of Nanoceram was determined to be 3.02%. Finally, successful detection of Norovirus GII in sewage from Yangqiao River, Fengtai District, Beijing was acheived. All these data had shown that the Naneceram filter concentration method could concentrate Norovirus from environmental water with a steady effects.
Filtration ; instrumentation ; methods ; Fractional Precipitation ; instrumentation ; methods ; Genotype ; Norovirus ; classification ; genetics ; isolation & purification ; Rivers ; virology ; Water Microbiology

OBJECTIVETo investigate prevalence of human immunodeficiency virus (HIV) infection and risk factors for its transmission among injection drug users (IDUs) in Liangshan Yizu Autonomous Prefecture of Sichuan Province, China.

METHODSA community-based survey was conducted to investigate demographic characteristics, pattern and frequency of sharing injection equipment, and sexual behaviors in IDUs. Blood samples were also collected from them to detect for antibodies against HIV and syphilis.

RESULTSA total of 379 subjects were recruited with informed consent for study through community outreach and peer recruiting methods. Their prevalence of HIV infection was 11.3% (43/379). Ethnicity, frequency of sharing syringes and cotton swab during the past three months and syphilis infection associated with HIV infection by univariate analysis using chi-square test. Multivariate logistic regression analysis showed odds ratio of frequency of sharing syringes for HIV infection during the past three months was 2.28 (95% CI 1.18 - 4.43), and that for syphilis infection 3.10 (95% CI 1.48 - 6.48).

CONCLUSIONFrequency of sharing syringes during the past three months associated with syphilis and HIV infection.

Adolescent ; Adult ; China ; epidemiology ; Disease Transmission, Infectious ; Female ; HIV Infections ; epidemiology ; transmission ; Humans ; Logistic Models ; Male ; Needle Sharing ; adverse effects ; Prevalence ; Sexual Behavior ; Substance Abuse, Intravenous ; virology