ObjectivesTo investigate the effects of 5-aza-2-deoxycytidine(5-Aza-dC),a methylation inhibitor,on the expression and methylation of tissue factor pathway Inhibitor (TFPI-2) gene in PANC1 cell line of pancreatic cancer.MethodsPANC1 cell lines were treated with different dosages of 5-Aza-dC ( 1× 10-7,5 × 10-7,1× 10 -6 mol/L).The status of TFPI-2 methylafion and expressions of TFPI-2 mRNA and protein were determined by MSP,RT-PCR,and Western blot.Results TFPI-2 gene CpG island was completely methylated,and there was no expression of TFPI-2 mRNA and protein without 5-Aza-dC treatment.After treatment with different dosages of 5-Aza-dC( 1 × 10-7,5 × 10 -7,1 × 10-6 mol/L),TFPI-2 gene CpG hypermethylation was reversed from incomplete methylated to complete non-methylated.The relative expressions of TFPI-2 mRNA were 0.211± 0.087,O.327 ± 0.068,0.609 ± 0.017; and the relative expressions of TFPI-2 protein were O.429 ± O.121,O.675 ± O.044,1.132 ± O.124 in a dose-dependent manner ( P ＜0.05 ).ConclusionsThe hypermethylatien of promoter region may be the primary reason for TFPI-2 gene expression down-rogtdation and inactivation.5-Aza-dC may reverse the hypermethylation of TFPI-2 gene,and induce the m-expression of TFPI-2 mRNA and protein.

Objective To observe the effect of Ginkgo biloba extract (EGb) on intestinal function after spinal cord injury (SCI) in rats. Methods 36 Sprague-Dawley rats were randomly divided into group A (n=12), group B (n=12) and group C (n=12). SCI model was established with Allen's mode (10 g×25 mm) at T10. 30 minutes later, group A was intraperitoneally injected with methylprednisolone 30 mg/kg every 24 hours; group B was injected with Shuxuening injection (EGb) 1.75 mg/kg every 24 hours; group C were injected with equal volume of saline. The slow wave of intestinal smooth muscle was measured, the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in serum were determined 1 day, 3 days and 7 days after modeling, while intestinal tissue was tested with HE staining. Results The amplitude and frequency of the myoelectric slow wave increased in the groups A and B 3 and 7 days after modeling compared with those in the group C (P<0.05); meanwhile, the activity of SOD increased and content of MDA decreased in the groups A and B (P<0.05). The HE scores decreased in the groups A and B compared with those in the group C (P<0.05), which presented that the inflammatory exudation was mild, the hemorrhagic spot was few and the area was limited. The intestinal villous of the group C was blunt with large infiltration of inflammatory cells and inflammatory exudate on the mucosal surface. Conclusion EGb can improve the recovery of intestinal function in rats spinal cord injury through antioxidant.

Objective To evaluate the importance of plasma MicroRNAs in early diagnosis of acute myocardial infarction (AMI).Methods 24 patients with AMI as the test group and 20healthy volunteers as the control group were enrolled in this study.Plasma levels of microRNA-1,microRNA-133a,microRNA-208a and microRNA-499 were detected by quantitative real time polymerase chain reaction (qRT-PCR) at 3 h,6 h,12 h,24 h,48 h,72 h after the onset of AMI.Results Plasma microRNA-1 level was greatly increased and reached the peak at 3 h after AMI,then was decreased gradually to normal level at 72 h after AMI.Plasma microRNA-133a level was significantly elevated at 6 h after AMI,reached peak at 12 h after AMI,then was decreased to normal level at 48 h after AMI.Plasma microRNA-1 and microRNA-133a levels were correlated with cTnI expression.The peak time of microRNA-1 was earlier than that of cTnI,while the peak time of microRNA-133a was the same as that of cTnI.Conclusions Increased circulating microRNA-1 and microRNA-133a may serve as potential and novel biomarkers for early diagnosis of AMI.

To observe the effect of electro-acupuncture on matrix metalloproteinase in degenerated cervical intervertebral disc rats.Methods:The rat model of cervical intervertebral disc degeneration was induced by unbalanced dynamic and static forces,then the rats were randomly allocated to model group,electro-acupuncture groupⅠ(acupoints Dazhu(BL 11) and Tianzhu(BL 10) were electro-acupunctured),and electro-acupuncture group Ⅱ(acupoints Dazhu (BL 11),Tianzhu(BL 10)and Shenshu(BL 23) were electro-acupunctured),with the normal rats as control.then the HE staining method was adopted to observe the morphological change of cervical intervertebral disc and the immunohistochemical staining method was used to detect the marx metalloproteinases-1(MMP-1) and matrix metalloproteinases-3(MMP-3) in cervical intervertebral disc.Results:The morphological observation showed that after electro-acupuncture treatment,the degrees of cervical intervertebral disc degeneration in electro-acupuncture groupⅠand electro-acupuncture group Ⅱ were alleviated,and the electro-acupuncture group Ⅱ was superior to electro-acupuncture group Ⅰ.The expressions of MMP-1 and MMP-3 in degenerated cervical intervertebral disc were increased(P＜0.01),and after electro-acupuncture treatment,the expressions of MMP-1 and MMP-3 in degenerated cervical intervertebral disc were decreased,especially the electro-acupuncture group Ⅱpresented the better effect as compared with electro-acupuncture group Ⅰ(P＜0.05).Conclusions:Electro-acupuncture at acupoints Dazhu (BL 11) and Tianzhu (BL 10) had a certain therapeutic effect on cervical intervertebral disc degeneration,and in combination with acupoint Shenshu(BL 23),this therapeutic effect could be enhanced,its action mechanism might be that electro-acupuncture can reduce the expressions of MMP-l and MMP-3 in degenerated cervical intervertebral disc,inhibit the degradation of matrix in intervertebral disc.so as to relieve cervical intervertebral disc degeneration.

Purpose To explore the biological significance of BCL-6 and ZEB2 in invasion,metastasis and prognosis of breast cancer.Methods The expressions of BCL-6,ZEB2 protein and mRNA were detected respectively in 228 cases of breast cancer and 80 cases of breast benign lesions by immunohistochemical SP two-step staining and situ hybridization.Result The expression levels of BCL-6,ZEB2 protein and mRNA in breast cancer tissues were significantly higher than in breast benign lesions (P ＜ 0.05).The expressions of BCL-6 was positively correlated with tumor size,lymphatic metastasis,histological grade,TNM staging and HER-2 expression (P ＜ 0.05).The expressions of ZEB2 was positively correlated with tumor size,lymphatic metastasis,TNM staging and HER-2 expression (P ＜ 0.05).The overall survival and relapse-free survival of BCL-6 and ZEB2 positive expression were significantly less than the negative expression (P ＜ 0.01).Conclusion The BCL-6 and ZEB2 are closely correlated with the evolution process of breast cancer,which may become important means for monitoring and warning the metastasis,invasion,and prognosis of breast cancer.

Objective To establish the a staging system based on scoring of ketamine associated urinary dysfunction (KAUD) and evaluate its diagnosis performance and clinical value.Methods A retrospective analysis of 135 admitted KAUD patients from September 2009 to September 2015 was performed.All patients were classified into three stages according to bladder and upper urinary tract impairment, and treated with subsequent strategy.The behavioral modification and pharmacotherapy was performed in stage Ⅰ which image investigation was normal.Hydrodistension and intravesical perfusion was used if pharmacotherapy failed in stage Ⅱ patients who had bladder impairment in image or urodynamics.Augmentation cystoplasty or cystectomy with urinary diversion was optional if aforementioned treatment failed in stage Ⅲ who had upper urinary tract involvement and abnormal renal function.The voided volume, micturition interval, and nocturnal void frequency as recorded in their 3-day bladder diary, PUF, ICPI and ICSI score after treatment in one month, were compared with their baseline data prior to treatment.Ketamine abusing history, renal and liver function, bladder and upper urinary tract changes, urodynamics and radioisotope renography correlation with stages were analyzed.A scoring system was established and the concordance between the clinical staging and model staging of each patient was compared.Receiver operating characteristic curve was employed to testify the sensitivity and specificity.Results There were 39, 80 and 16 patients included in each stage respectively.After subsequent strategy of treatment, voided volume, micturition interval, and nocturnal void frequency, PUF, ICPI and ICSI were improved significantly.Age (P =0.018), ketamine abusing history (P =0.006), eGFR (P =0.001), ALT (P =0.037), γ-GT (P ＜ 0.001), bladder (P =0.013) and ureteral changes (P ＜ 0.001), hydronephrosis (P ＜ 0.001) and radioisotope renography abnormality (P =0.01) were correlated with clinical stages.In total, 107 cases' stages based on scoring were accordant with clinical stages (30, 65 and 12 cases in each stage respectively).AUC of the scoring model ROC was 0.922 on cut point between Ⅰ and Ⅱ stages and AUC of the scoring model ROC was 0.978 on cut point between Ⅱ and Ⅲ stages.Conclusions We presented a scoring staging system of KAUD based on ketamine abusing history, laboratory test results, and imaging findings.It might be helpful to evaluate the progression on bladder and upper urinary tract, and provide a reference for treatment.This staging system based on scoring still needs further validation and improvement.

Objective To identify the risk factors of chronic ketamine-induced upper urinary tract dysfunction (KUUTD).Methods Medical records of 103 patients with ketamine-associated urinary dysfunction(KAUD) from Sep.2009 to Jul.2015 were retrospectively reviewed.All the patients were divided into the upper urinary tract dysfunction group(n =39)or the normal group(n =64)according to the diagnostic criteria.Indexes such as general features (age,gender,BMI),drug-abusing characteristics (abusing time,frequency,dose,abstinent period),duration of LUTS,pelvic pain and urgency/frequency symptom score(PUF score),serum parameters (WBC,ALT,AST,γ-GT),bladder diary (daytime inter-void interval,nocturia,average urine volume),urodynamic data (maximum cystometric capacity,post-void residual,maximum urethral closure pressure,maximum and average flow rate,instable contraction,bladder compliance) and imaging manifestations were compared between the two groups.Logistic regression analysis was performed for the evaluation of variables with statistical significance.Results There was significantly difference between the two groups in BMI (t =2.127,P =0.043),drug-abusing frequency (t =2.335,P =0.029),abstinent period (t =3.451,P =0.003),duration of LUTS (t =2.289,P =0.036),PUF score (t =3.030,P =0.004),γ-GT value(t =2.697,P =0.012),MCC(t =5.056,P ＜ 0.001),MUCP(t =3.015,P =0.008),BC (t =8.562,P ＜ 0.001) and imaging findings of bladder contracture (x2 =15.155,P ＜0.001).The multivariate Logistic regression analysis showed that drug-abusing frequency(OR =1.279,P =0.038),abstinent period(OR =0.394,P =0.028),γ-GT value (OR =1.007,P =0.032),MCC (OR =0.775,P =0.011)and BC (OR =0.013,P =0.008)were the key factors of KUUTD.Conclusion Consecutive and frequent drug abuse,combination with chronic obstructive cholangitis,reduced capacity and low compliance bladder are the risk factors of upper urinary tract dysfunction associated with ketamine abuse.

To systematically evaluate the efficacy and safety of tripterygium glycosides (TG) combined with ACEI/ARB preparation in treating diabetic nephropathy stage IV. The computer retrievals were made in Cochrane Libarary, PubMed, Embase, SCI, Sinnomed, CNKI, Chinainfo and VIP, and hand retrievals were conducted for meeting and academic papers (updated to December 30, 2014), in order to collect randomized controlled trials and quasi-randomized control trials for TG combined with ACEI/ARB preparation in treating diabetic nephropathy stage IV and set the literature inclusion and elimination standards. Eligible literatures were included and evaluated according to standards in Cochrane Handbook. RevMan 5.3 and Stata 12.0 were used for a Meta-analysis. A total of 13 randomized controlled trials and quasi-randomized control trials involving 1119 patients with diabetic nephropathy were included. The Meta analysis result showed that compared with the control group, the combination group showed better effects in reducing the 24-hour urinary protein [MD = -0.84, 95% CI (-1.02, -0.66)], raising albumin [SMD = 0.98, 95% CI (0.81, 1.16)], the total efficiency [OR = 4.23, 95% CI (2.77, 6.46)] and the significant efficiency [OR = 5.35, 95% CI (2.70, 10.60)], with no statistical difference in Serum Creatinine between Both groups [MD = -0.82, 95% CI (-4.30, 2.66), P = 0.64]. However, the risk of adverse reactions increased by 7% [RD = 0.07, 95% CI (0.03, 0.12)]. The Egger's test showed no publication bias. Tripterygium Glycosides combined with ACEI/ARB in treating diabetic nephropathy stage IV is supper than the single administration of ACEI/ARB, with a good prospect in clinical application. Nevertheless, due to the small-size and low-quality samples in this study, more high-quality and large sample-size randomized controlled trials shall be conducted to verify the findings.
Angiotensin Receptor Antagonists ; administration & dosage ; Angiotensin-Converting Enzyme Inhibitors ; administration & dosage ; Diabetic Nephropathies ; drug therapy ; Drug Therapy, Combination ; Glycosides ; administration & dosage ; Humans ; Tripterygium ; chemistry

Objective:To construct a DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the anti-HBV siRNA genes, and to express this DNA vaccine in HepG2 cells. Methods:The HBV multi-epitope antigen gene was designed and synthesized before it was fused with enhanced green fluorescent protein(EGFP) gene, and cloned into the multi-clone site(MCS) of the eukaryotic expression vector pVAX1. The expressinig units of hIL-12 and siRNA were cloned into the BspH I and Mlu I site of pVAX1 respectively. Then the recombinant plasmid pVAX1-siHBV-HB-EGFP-hIL12 was transiently transfected HepG2 cells. The expression of HBV compound multi-epitope gene was observed through EGFP report gene. The expression of hIL-12 was analyzed by ELISA and the effects of anti-HBV siRNA was confirmed with rtPCR . Results: The analysis of enzyme digestion and sequencing both demonstrated that the trible-expressing HBV DNA vaccine has been constructed successfully. The green fluorescent image was detected in the transfected cells which could confirm the expression of the multi-epitope antigen gene. The amount of hIL-12 secretion was 1289pg/ml in supernatant at 48h after transfection and 1712pg/ml at 72h after transfection. The mRNA amount of HBV S gene, which was the siRNA target, had been obviously knockdown. Conclusion: The DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the siRNA genes was constructed and transiently expressed in HepG2 cells, and siRNA had shown us a good anti-HBV effect. It laid a foundation of further study on anti-HBV effect of the new DNA vaccine.

AIM: To investigate the safety and effect of EX-PRESS glaucoma drainage device on open-angle glaucoma.
METHODS: A retrospective review of 40 patients ( 47 eyes ) whose eyes were diagnosed as open angle glaucoma through the best - corrected visual acuity ( BCVA ) , intraocular pressure ( IOP ) , gonioscope, examination of the ocular fundus, and visual fields et al. Fourty patients 6-73y old ( mean 39. 90±16. 50y old) with the BCVA from 0. 01 to 0. 6 and the IOP from 18mmHg to 65mmHg were treated with EX-PRESS glaucoma drainage device after achieving not well IOP over three drugs. Follow - up of these patients 1d, 3d and 1wk after treatment, IOP, slit lamp examination were retrospectively observed. The changes of the BCVA, IOP were used to evaluate the safety and effect before and after treatmeat.
RESULTS: The mean BCVA was 0. 26 ± 0. 29 pre -operatively, while it was 0. 24±0. 22 after treatment one week. The mean BCVA decreased slightly, but there was no statistic difference between pre-treatment and post-treatment (t=1. 56,P=0. 13). The mean IOP was (36. 62 ±14.01)mmHg pre-operatively, (10.04±5.77)mmHg after treatment one day , ( 9. 59±4. 93 ) mmHg after treatment three days, (9. 47±3. 06)mmHg after treatment one week. The IOP was decreased significantly in post-treatment after one day, three days, one week compared with pre-treatment (F=157. 20, P<0. 05). Except for 5 eyes of the IOP below 5mmHg, there was no ocular or systemic adverse events observed in all patients.
CONCLUSION:EX-PRESS glaucoma drainage device is an effective and safe treatment for the patients with open-angle glaucoma. The risk and complication are low intraoperatively and postoperatively.