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 examine the expression and function of thrombonspondin-1 (TSP-1) in oxygen-induced retinopathy in new-born mice, and to investigate its role in retinal neovascularization. Methods:A total of 40 C57BL/6J newborn mice were divided equally into a model group (n=20)andanormalcontrolgroup(n=20).Miceinthemodelgroupwereexposedto75%oxygentoestablish the oxygen-induced retinopathy (OIR) model. On the 7th, 9th, and 11th day after the birth of mice, 5 mice were randomly selected each time from the 2 groups to examine the expression of TSP-1 mRNA with reverse transcription polymerase chain reaction (RT-PCR). After that, 5 mice were selected on the 11th day to observe the retinal neovascularization by fluorescein angiography retinal flatmount. Results:On the 11th day, fluorescein angiography retinal flatmount showed that the retinal blood vessels presented mean network distribution in the normal control group, while in the model group, a lot of dilatated areas in the retinal main vessels surrounded the optic disc. Meanwhile lots of new blood vessels were found surrounding the optic disc with irregular distribution but well distributed peripherial retinal small vessels, which was typical of early stage OIR. There was no signiifcant difference in the retinal TSP-1 mRNA level between the model group and the normal control group in the postnatal 7-day mice (P>0.05). Compared with the normal control group, the expression of TSP-1 mRNA in the model group was signiifcantly lower in postnatal 9-day and 11-day mice (P<0.05, P<0.01) , and the expression of TSP-1 mRNA in postnatal 9-day mice was lower than that in the postnatal 11-day mice(P<0.05).
Conclusion:In the early stage of OIR model (retinal vascular growth and development stage), the expression of TSP-1 mRNA in the retinal tissue is gradually decreased, implying that TSP-1 (as a negative regulatory factor) may be involved in the formation of retinal neovascularization in the early stage.

Objective To compare the detection performance of the modified serum test (TRUST) method and the traditional method in syphilis screening.Methods A series of TRUST high titer syphilis serum was diluted,and the positive rate of each method was calculated by using the improved method and the traditional method.Comparison of two detection methods of C50,C5 ～ C95 interval,as well as the accuracy of the density curve,and the consistency of the two methods were compared,and diagnostic performance were compared.Results The improved method of C50 was less than the traditional method of C50,and the improved method of C5 ～ C95 range was narrower than the traditional method,compared with the traditional method.The improved method of the non precision density curve was steeper than the traditional method,and the two confidence interval of the consistency degree of the 95％ methods was 73.4％ to 95.8％.The diagnostic sensitivity (SEN),diagnostic specificity (SPE),positive predictive value (PPV),negative predictive value (NPV) and diagnostic efficiency(DF) of the improved method were 64.29 ％,99.1％,85.71％,97.05 ％ and 96.39 ％,respectively.The SEN,SPE,PPV,NPV and DF of the traditional methods were 3.75 ％,98.49 ％,75 ％,96.18 ％ and 95 ％,respectively.The improved method was superior to the traditional methods in the two aspects of SEN and PPV (x2 =8.25,10.03,all P＜0.05),with the statistically significant difference.The improved method was slightly higher than the traditional method in SPE,NPV and DF (x2 =2.39,3.45,4.03,all P＞0.05),with the no statistically significant difference.Conclusion The precision,diagnostic sensitivity and diagnostic specificity of the improved method was higher than that of the traditional method,and it can be applied to the detection of large batch samples with the aid of the full automatic enzyme immunoassay instrument.The improved method can be used to replace the traditional method for syphilis screening.

Objective To investigate the potential role of IC0s-Ig and CsA in inducing transplantation tolerance and the mechanisms thereof. Methods ICOS-Ig was a fusion protein of human ICOS extracellular region and IgG Fc fragment.Cardiac allograft from BALB/c mouse was transplanted to C57BL/6 mouse Animals were randomly divided into 5 groups:(1) un-treated group;(2)IgG-treated group,250g,i.p.day 2,4,6;(3)ICOS Ig-treated group,250g,i.P.day 2,4,6;(4) CsA-treated group,10 mg/kg,i.p.day 0-6;(5)ICOS-Ig+CsA-treated group.The survival time and pathological changes of the cardiac allografts were monitored.The mixed lymphoeyte reaction (MLR) and the alloantibody level of the recipients were also detected.Results The median survival time (MST) of the cardiac allografts was (8.5±1.5) days in un-treated group,(8.0±0.8) days in IgG-treated group,(29.5±7.7) days in ICOS-Ig-treated group,and(21.0±5.0) days in CsA-treated group.respectively.In ICOS-Ig + CsA-treated group,the MST was prolonged to longer than 100 days,which was significantly longer than other four groups(P<0.01).Allogeneic hearts from ICOS-Ig and/or CsA immunized recipients revealeel milder histological changes than control groups(P<0.05).Mechanical ahalysis revealed that splenic T cells from recipients also exhibited depressed MLR activities.The alloantibody level in ICOS-Ig-treated group and/or CsA-treated group was lower than in control groups(P<0.05),suggesting ICOS-Ig not only inhibited cell immunity,but also depressed humoral response.Conclusion ICOS-Ig combined with CsA leads to a long-term survival of mouse cardiac allografts.The induced tolerance is donor-specific and the mechanisms may be associated with T cell anergy.

Objective To explore the effect of dexmedetomidine combined with ropivacaine for continuous lumbar plexus block on the quality of early recovery in patients undergoing total hip ar-throplasty.Methods Sixty patients (35 males and 25 females)with ASA physical status Ⅱ or Ⅲ, aged 65-84 years,undergoing total hip replacement were randomly assigned to dexmedetomidine com-bined with ropivacaine group (group D)or ropivacaine group (group C).All patients received ropiva-caine which was administered via continuous lumbar plexus block as patient-controlled analgesia (PCA)after surgery.The PCA were programmed with a background infusion 8 ml/h of ropivacaine, bolus dose was 4 m1 and the block time was 30 min;the patients in group C received 0.2% ropiva-caine,and the patients in group D received 0.1% ropivaciane combined with 1 μg/ml of dexmedeto-midine by the way of intravenous infusion.In addition,all patients received another patient-controlled intravenous analgesia (PCIA)with 1 mg/ml of morphine for relieving the explosive pain.The PCIA was programmed with a lobus dose of morphine 1 mg without background dose,the block time was 5 min.Consumption of morphine and visual analog scale (VAS)score,muscle strength and maximum flexion and abduction of hip joint were recorded at the time points of 6,12,24 and 48 h after opera-tion.The side-effect reactions such as nausea,vomiting,drowsiness and itching were recorded.The sleep quality was assessed with Pittsburgh sleep quality index (PSQI)on day 1 before operation,day 1 and day 7 after operation.Results The consumption of morphine in group D was significantly fewer than group C (P <0.05).Compared with group C,muscle strength and maximum flexion and abduc-tion of hip joint in group D were increased at 6,12,24 and 48 h after operation (P <0.05 ).PSQI scores were decreased on the day 1 and day 7 after operation.The incidence of delirium,nausea and vomiting,drowsiness and itching in group D were decreased (P <0.05).Conclusion 0.1% ropiva-ciane continuous lumbar plexus block combined with 1 μg/ml of dexmedetomidine by the way of in-travenous infusion could provide satisfied analgesia for the operation of total hip arthroplasty;the sleep quality is improved,the functional exercise of hip joint is promoted.

Objective To study the effect of field long march comprehensive training on freshmen' mental quality and mental health in military university. Methods Mental quality questionnaire for army men (MQQA), symptom checklist 90 (SCL-90), self-rating anxiety scale (SAS), self-rating depression scale (SDS) and trait anxiety inventory (TAI) were carried out on 330 freshmen at various stages on their military training. All data were analyzed by paired-sample t test. Results ①As compared with pre-training, all SCL-90 factor scores except depression factor decreased significantly (P<0.05); compared to barracks military training, recruits revealed a decrease (P<0.05) in all SCL-90 factors except factors on fear, paranoia and mental disorder. ②The scores of SDS and SAS displayed a“V”shape as they decreased after the early rise at the course of military training. After the barracks military training, SAS and SDS scored high (P<0.01);Compared with pre-training, TAI scores dropped significantly (P<0.01) after field long march; compared with post barracks military traning results, SAS, SDS and TAI scores of recruits after field long march decreased significantly (P<0.05). ③After barracks military training and field long march comprehensive training, recuits' mental quality factors scored no significant distinction (P>0.05). Conclusion Filed long march effectively improved mental health of military university recruits and relived their anxiety as well as depression.

Objective To observe effects of electro-acupuncture (EA) on the expression of apoptosis related protein B-cell lymphoma/leukemia gene 2 (Bcl-2) and Bcl-associated protein X (Bax) in the hippoeampal CA1 region of rats with cerebral infarction,and on their behavior. MethodsAn animal model of cerebral ischemia was established by right side middle cerebral artery occlusion using thread in 48 male,adult Wistar rats,who were then randomly divided equally into an EA group and a control group.Both groups were sub-divided into 1 week,2 weeks and 3 weeks subgroups.The EA group began receiving EA 24h after the occlusion,applied at the Baihui (DU20) and Dazhui (BU14) points,once daily,for one,two or three weeks.The control group was reared conventionally and was not given any special treatment.The rats'learning and memory,motion and neural function were evaluated.The expression of Bcl-2 and Bax in the CA1 region of the hippocampus on the infarcted side and changes of apoptosis indexes were detected using immunohistochemical techniques.ResultsThe learning,memory,motion and neural function of the EA group rats were better on average than those of the control group at all observation time points.The expression of Bcl-2 protein increased along with reduced expression of Bax protein in the EA group significantly more than among the controls.TUNEL positive cells in the CA1 region of the hippocampus were reduced significantly in the EA group compared with the control group.ConclusionsBehavioral ability after cerebral infarction can be improved by EA at the Baihui (DU20) and Dazhui (BU14) points.EA can up-regulate the expression of Bcl-2 and down-regulate the expression of Bax and reduce TUNEL positive cells in the CA1 region of the hippocampus after cerebral infarction,which might be the mechanism of its neuroprotection.

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 analyze the correlation of hypersensitive C-reactive protein(hs-CRP),homocysteine(Hcy) and D-dimer(D-D) with pathological change degree of coronary heart disease(CHD),and to investigate the diagnostic specificity and sensitivity of joint detection in CHD.Methods A total of 100 cases of CHD patients(experimental group) and 100 healthy subjects(control group) were enrolled,.Patients of the experimental group were divided into four groups on the basis of complications,including simple CHD group(32 cases),amalgamating hypertension group(46 cases),amalgamating diabetes group(9 cases),amalgamating hypertension diabetes group(13cases).The levels of hs-CRP,Hcy and D-D in the five groups were detected and analyzed.Results The levels of hs-CRP,Hcy and D-D in the experimental group were all higher than the control group(P<0.05).Logistic regression confirmed that three indexes were the independent risk factors for CHD.Each indicator has a certain clinical significance to the diagnosis and treatment of CHD but the value of Hcy could be better.Joint detection of hs-CRP and Hcy could be an ideal combination of detection,and the three joint detection might not be suitable for early diagnosis and treatment of CHD.The levels of hs-CRP and D-D in simple CHD group,amalgamating hypertension group,amalgamating hypertension diabetes group were all higher than amalgamating diabetes group(P<0.05).Conclusion hs-CRP,Hcy and D-D could be the independent risk factors of CHD,and joint detection might be with important clinical value for diagnosis of CHD.