Objective To investigate the expression and methylation status of HOXA7 gene in human pancreatic cancer cell lines, and to explore the relationship between them.Methods HOXA7 mRNA expression of human pancreatic cancer cell lines BxPC3, CFPAC1, PANC1 and SW1990was detected by RT PCR.Bisulfite sequencing PCR (BSP) and combined bisulfite restriction analysis (COBRA) was used to test promoter methylation status.All the cell lines were treated by 5-aza-2-deoxycytidine (5-aza-dC), and HOXA7mRNA expression, methylation status was detected before and after this treatment.Results HOXA7 mRNA was expressed in BxPC3, CFPAC1 and SW1990, while there was no expression of HOXA7 mRNA in PANC1.HOXA7 promoter methylation rates of CFPAC1, BxPC3, PANC1 and SW1990 were 93.16%, 90.65%,90.09% ,52.30%.HOXA7 promoter methylation rate of SW1990 was significantly lower than those in other 3cell lines ( P ＜0.01 ).After 5-aza-dC treatment, HOXA7 mRNA of PANC1 was expressed again, and HOXA7mRNA of BxPC3 was increasingly expressed;while the expression of HOXA7 mRNA in CFPAC1 and SW1990was not significantly changed after 5-aza-dC treatment.Conclusions The expression of HOXA7 mRNA in BxPC3 and PANC1 was closely correlated with promoter hypermethylation, while there was no obvious relation in CFPAC 1 and SW1990.

Objective To determine the methylation status and expression of Ras association domain family 1A (RASSF1A), and the possible effect between promoter aberrant methylation and pathogenesis of pancreatic cancer. Methods The methylation status of RASSF1A promoter CpG island (CGI) pancreatic cancer cell line BxPC3 was detected in 5 cases of normal pancreatic tissue and 13 pairs of pancreatic cancer tissues (tumor and peri-tumor) by using COBRA (combined bisulfite restriction analysis) and the methylation rate was calculated. The RASSF1A mRNA expression of BxPC3 was compared between pre- and post-treatment of the inhibitor of DNA methyltransferase (5-Aza-2-deoxycitydine, 5-Aza-dC). Results The average methylation rate of RASSF1A promoter CGI was 62.90% in BXPC3, 9.14% in normal pancreas, 53.79% in peri-tumors (TP), and 55.82% in tumors. The methylation rates in port-tumors and tumors were significantly increased when compared with that of normal pancreas (P < 0.01), while there was no significant difference between in peri-tumors and tumors (P > 0.05). After 5-Aza-dC treatting BxPC3 cells, the methylation rates decreased to 42.5% (P < 0. 05) and RASSF1A mRNA expression was enhanced. Conclusions Aberrant hypermethylation of RASSF1A promoter CGI could be considered as an early event in the process of pancreatic cancer and participates in the pathogenesis of pancreatic cancer.

Objective To investigate the diagnostic value of 1H magnetic resonance spectroscopy (1H-MRS) in mammary tumors and to discuss the technique factors which influence the detection rate.Methods The 1H-MRS features of 47 mammary tumors, of which 24 malignant tumors and 23 benign tumors confirmed by pathology were analyzed. All of the tumors were detected before Gd-DTPA enhancement. Results Eleven of 24 malignant tumors showed increased choline resonance peak at 3.24 ppm while 4 of 23 benign ones at 3.24 ppm .The positive value were 45.8% and 17.4% respectively. The sensitivity and specificity were 45.8% and 82.6% respectively by using 1H-MRS to discriminate benign from malignant tumors. The main factors influencing the detection rate were low suppressed lipid, low suppressed water and low single-noise rate.Conclusion Choline is not special features of malignant tumors. Choline can be obtained despite the nature of tumor if they grow rapidly. The low sensitivity of choline to be detected mainly dues to technique factors.

Objective To investigate the efficacy of skin scraping plus movable cupping in treating neck-shoulder syndrome. Method One hundred and eighty patients with neck-shoulder syndrome were randomly allocated to treatment and control groups. The treatment group received skin scraping plus movable cupping and the control group, acupuncture. Result The total efficacy rate was 88.8% in the treatment group and 77.5% in the control group; there was a statistically significant difference between the two groups (P<0.05). Conclusion Skin scraping plus movable cupping has a good therapeutic effect on neck-shoulder syndrome.

Objective To investigate the efficacy of dexmedetomidine combined with target-controlled infusion (TCI) of propofol and remifentanil for fiberoptic bronchoscopy in the elderly patients.Methods Forty ASA Ⅱ or Ⅲ patients,aged 65-75 yr,with body mass index of 20-30 kg/m2,scheduled for elective fiberoptic bronchoscopy,were randomly divided into 2 groups (n =20 each):control group (group C) and dexmedetomidine group (group D).In group D,a loading dose of dexmedetomidine 0.5/μg/kg was injected at 10 min before induction of anesthesia,followed by infusion at 0.5 μg· kg-1 · h-1 until the end of fiberoptic bronchoscopy.While the equal volume of normal saline was given instead in group C.Anesthesia was induced with TCI of propofol and remifentanil.The target effect-site concentration (Ce) of propofol was 3 μg/ml.When the plasma concentration and Ce were balanced,TCI of remifentanil (target Ce 4 ng/ml) was started.The fiberoptic bronchoscope was placed after consciousness was lost and then the Ces of propofol and remifentanil were adjusted to 1-3 μg/ml and 2-4 ng/ml,respectively.MAP,HR and OAA/S score were recorded before induction (T0),immediately after induction (T1),when the tip of fiberoptic bronchoscope reached the glottis (T2) and carina (T3),at the end of bronchoscopy (T4)and 10 min after the end of bronchoscopy (T5).The consumption of propofol and remifentanil,duration of bron-choscopy,emergence time,adverse cardiovascular events and side effects such as hyoxemia,nausea and vomiting,regurgitation and aspiration were recorded.Results Compared with group C,OAA/S score at T5 and the consumption of propofol and remifentanil was reduced,and emergence time was shortened,and the incidence of hypotension and hyoxemia was decreased in group D (P ＜ 0.05).No patients developed side effects such as hyoxemia,nausea and vomiting,regurgitation and aspiration in both groups.Conclusion Dexmedetomidine (infusion at 0.5 μg·kg-1 ·h-1 after a loading dose of 0.5 μg/kg) combined with TCI of propofol and remifentanil can be safely and effectively used for fiberoptic bronchoscopy in the elderly patients.

ObjectiveTo investigate the methylation of the promoter region in miRNA in pancreatic cancer cell line PANC1 and normal pancreatic tissue,to discover the miRNA with hypermethylation associated with pancreatic cancer.MethodsThe genomic DNA of PANC1 and normal pancreatic tissue was extracted,and fractured by ultrasound.Methylation DNA fragments were obtained by 5-methyl of pyrimidine nucleoside antibodies and immunomagnetic beads.The hypermethylation miRNA differentially expressed between PANC1 and normal pancreatic tissue was selected by using methylation DNA chip.BSP ( bisulfite genomic sequencing PCR) and TA clone sequencing was performed for further validation.The genomic DNA of pancreatic cancer cell lines BXPC3,CFPAC1,PANC1 and SW1990 was extracted.The COBRA (combined bisulfite restriction analysis) was used to validate differentially expressed hypermethylation miRNA.ResultsEight differentially expressed hypermethylation miRNAs were screened from the DNA methylation chips,then five of them were selected for sequencing.The methylation status of miRNA-615,-663,-663b was significantly higher in the PANC1 than in normal tissues (60.6％ vs 7.6％,88.8％ vs 22.2％,94.4％ vs 13.0％ ) ; the methylation status of miRNA-675 was not significantly different between PANC1 and normal pancreatic tissue (76.0％ vs 100％ ).Due to large error in sequencing,miRNA1826 was excluded.The results of COBRA confirmed all the 4 miRNAs were highly methylated in PANC1 ; except for miRNA-675,other 3 miRNAs were highly methylated in BxPC,miRNA-663,miRNA-663b were highly methylated in CFPAC1,while miRNA-615,miRNA-663 were highly methylated in SW1990.ConclusionsHypermethylation miRNAs were differentially expressed between pancreatic cancer cell lines and normal pancreatic tissue,among them,highly methylated miRNA-663 was possibly associated with pancreatic cancer.

Objective To study the effects of low-frequency electrical stimulation(LFES)on motor function and the expression of glia fibrillary acidic protein(GFAP)around cerebral infarction sites in rats.Methods Fifty-four male adult Sprague-Dawley rats were randomly divided into a LFES group,a placebo group and a sham operation group(18/group).All groups were randomly divided into 3 treatment groups.A rat model of middle cerebral artery occlusion(MCAO)was established using intraluminal filament occlusion.Treatment was carried out 3 d after the operation.Rats in the LFES treatment groups were stimulated with LFES for 3,7 or 14 days (10 min/d);the placebo groups were treated in the same way without electric stimulation;the sham operation subgroups didn't receive any therapy.Scores on a beam-walking test,a rotating pole test and a screen test were assessed at each time point mentioned above.Expression of GFAP was also assessed using immunohistochemcal techniques.Results The paralysed limbs recovered motor function better in the LFES groups than in the control groups.GFAP-positive cells were more numerous at the margins of the infarction area in the treated groups than in the control groups.Conclusions LFES might increase the expression of GFAP,which might be an important mechanism in improving brain plasticity after cerebral ischemia,aiding the recovery of the central nervous system and rebuilding its functioning.

Objective To explore the dynamic changes of von willebrand factor(VWF)and Pseleetin in the finger-replanted patients,and the relationship between the prognosis of the surgery and hypercoagulability.Methods From December 2004 to December 2006,eishty finger-replanted patients were recruited to our study.with 40 healthy volunteers as controls.Plasma VWF and P-selectin were detected by enzyme-linked immunosorbent assay(EUSA)in both controls and patients before or after replantation.Results The VWF and P-selectin levels had significant differences between the replantations and controls(F=14.76,11.76,P＜0.01).The VWF levels in the patients of 1,4,8,16 hours after replantation were(1 715±493),(1 396±549),(1 266±504),(1 163±436)U/L respectively,all markedly higher than the controls(P＜0.01).The P-selectin levels in patients of 1,4,8,16,24 hours after operation were(14.7±2.6),(12.5±3.0),(11.8±3.2),(11.1±3.0)、(10.5±2.6)μg/L,significanfly higher than the controls(P＜0.01).The VWF levels in patients of pre-replantion and the 1,4,8,16,24,48,72 hours after replantation were(854±209),(1 535±389),(1 177±407),(1 040±283),(958±216),(829±193),(777±151),(713±137)U/L in successful group,and were(1 202±164),(2 333±243),(2 146±161),(2 039±244),(1 865±170),(1 645±283),(1 427±331),(1 188±262)U/L in unsuccessful groups.They were all significantly different at the same test-time points between two groups(t=4.44,5.12,6.10,8.43,10.17,8.85,5.10.4.61,P＜0.05).The P-selectin levels in patients of 1,4,8,16,24,48,72 hours after replantation were(13.9±2.5),(11.2±2.0),(10.2±1.6),(9.6±1.2),(9.2±0.9),(9.5±0.6),(9.3±0.4)μg/L in successful group,and(17.2±1.0),(16.9±1.0),(17.0±1.3),(16.1±1.1),(14.9±1.5),(13.8±1.4),(12.8±1.2)μg/L in unsuccessful group.Significant difference existed at the same testtime points between two groups again(t=5.22.9.91,10.35,12.79,9.46.9.45,9.33,P＜0.01).After replantation,both VWF and P-selectin were rapidly elevated and went to the summit 4 hours later,then declined to pre-replantation level about 24 to 48 hours later after replantation.Conclusions VWF and P-selectin were associated with the hypercoagulability.Dynamic monitoring VWF and p-selectin may be useful in determining the existence of hypercoagulability and the therapy of anti-coagulability.

Objective To observe the clinical efficacy of heat-sensitive moxibustion plus point-toward-point needling in treating poststroke strephenopodia.Method Eighty patients with poststroke strephenopodia were randomized into a treatment group intervened by heat-sensitive moxibustion plus point-toward-point needling and a control group intervened by rehabilitation, 40 cases in each group. In addition to the basic treatment, the treatment group was given heat-sensitive moxibustion plus point-toward-point needling, and the control group was given rehabilitation treatment. Holden's Functional Ambulation Classification (FAC), Fugl-Meyer Assessment (FMA) of the lower-limb motor function, and Tinetti Gait Assessment (TGA) were adopted for evaluation of the two groups, and the clinical efficacies were compared.ResultThe effective rate was 90.0% in the treatment group versus 77.5% in the control group, and the difference was statistically significant (P<0.05); after the treatment,there was a significant difference in comparing the Holden's FAC between the two groups (P<0.05); the FMA score changed significantly after the treatment in both groups (P<0.05), and there was a significant difference in comparing the FMA score between thetwo groups after the treatment (P<0.05); the TGA score changed significantly after the intervention in both groups (P<0.05), and there was a significant difference in comparing the TGA score between the two groups after the intervention (P<0.05).ConclusionHeat-sensitive moxibustion plus point-toward-point needling can produce a significant efficacy in treating poststroke strephenopodia, as it can enhance the effective rate and improve the lower-limb motor function.

Objective: Comparison of alkaloid and polysaccharides content in different parts of whole plant of wild aconitum in Nanyang, and to explore the medicinal value of aboveground part and fibrous root of wild aconitum in Nanyang. Methods: The contents of three monoester alkaloids, three diester alkaloids in wild aconitum, radix aconiti carmichaeli, fibrous root and aboveground parts in Nanyang were determined by HPLC method; The content of crude polysaccharides was determined by sulfuric-phenol method, crude polysaccharides content from different tissues of wild aconitum in Nanyang in different harvest time was compared, including wild aconitum in Nanyang, wild radix aconiti carmichaeli in Nanyang, fibrous root, aboveground part. Results: Among the 4 parts of the wild Aconitum in Nanyang, the contents of 3 mono ester alkaloids rang from high to low: wild aconitum in Nanyang＞ wild radix aconiti carmichaeli in Nanyang＞ fibrous root＞ aboveground part (P ＜ 0.05), the contents of three diester alkaloids rang from high to low: wild radix aconiti carmichaeli in Nanyang＞ fibrous root＞ wild aconitum in Nanyang＞ aboveground part (P ＜ 0.05), the order of total alkaloid content from high to low is: wild radix aconiti carmichaeli in Nanyang＞ fibrous root＞ wild aconitum in Nanyang＞ aboveground part (P ＜ 0.05), The content of crudepolysaccharides in different tissues of wild aconitum in Nanyang were in the sequence of wild radix aconiti carmichaeli in Nanyang＞ wild aconitum in Nanyang＞ aboveground part＞ fibrous root (P ＜ 0.05) . Conclusion: In this study, an attempt was made to explore the differences of 6 kind salkaloid content in different parts of whole plant of wild aconitum in Nanyang, which provided scientific basis for the medicinal value of the fibrous roots and the aboveground parts.