Cytosine methylation is one of the major types of DNA epigenetic modifications and plays an important role in maintaining normal cell function and regulating gene expression. Bisulfite sequencing PCR (BSP) based cloning and sequencing is a general method for detecting DNA methylation at specific sites, which can clarify the methylation status of each CpG site in the target fragment. However, this method requires large amounts of single-clonal sequencing, which is complicated to operate, time consuming and expensive. Therefore, the development of an accurate, efficient and convenient DNA methylation detection technology is of great significance to improve the efficiency of epigenetic research. Based on the high-throughput mutation detection platform Hi-TOM (high-throughput tracking of mutations) developed by our group, we further established a site-specific DNA methylation high-throughput detection platform Hi-Meth (High-throughput Detection of DNA Methylation). After bisulfite treatment of DNA samples, the specific site-specific DNA methylation analysis results could be obtained through the Hi-Meth platform by performing only one round of PCR amplification. Using the Hi-Meth platform, the DNA methylation status of two promoter regions of rice were detected. The DNA methylation results from Hi-Meth were consistent with the results from BSP-based method. Thus, site-specific DNA methylation analysis results could be obtained accurately and conveniently through the Hi-Meth platform. In conclusion, Hi-Meth provides an important methylation detection platform for specific DNA regions, which has important significance for epigenetic research.
DNA Methylation ; Epigenesis, Genetic ; Epigenomics ; High-Throughput Nucleotide Sequencing/methods* ; Polymerase Chain Reaction ; Sequence Analysis, DNA/methods*

Objective To explore the perioperative management and surgical outcomes of fracture-dislocation of the upper cervical spine complicated with vertebral artery injury.Methods We retrospectively analyzed the clinical data of 19 patients diagnosed with fracture-dislocation of the upper cervical spine complicated with vertebral artery injury who had been treated at our department from January 2008 through December 2012.They were 14 males and 5 females,aged from 22 to 53 years (mean,35.2 years).All the patients had fractures of the atlas and/or axis,and disordered atlanto-axial relationship as well.According to Frankel grading system,8 cases were Grade D and 11 Grade E.The cervical posterior fixation with pedicle screws was adopted for all and hemostasis was conducted for those with bleeding from the broken vertebral artery.Their Frankel grade,Japanese Orthopedic Association (JOA) score and visual analogue scale (VAS) score were compared between preoperation and final follow-up.Results Intraoperative hemorrhage occurred in 2 patients from the injured vertebral artery.Hemostasis was achieved through direct tamponade with bone wax and gelatin sponge in one and through endovascular intervention after bleeding control by direct tamponade in another.Neither of them presented with symptoms of posterior circulation ischemia after operation.The mean operation time was 153.5 min,and the mean blood loss was 542.1 mL.All the patients were followed up for an average time of 28 months.Bony union was obtained in all after an average time of 13.5 weeks.Follow-ups revealed no ischemic stroke in this series.Symptoms of transient ischemic attack,like transient dizziness and blurred vision,appeared in 2 patients.At the final follow-up,all the patients were assessed as Frankel Grade E.Their mean JOA and VAS scores were significantly improved from 8.1 ± 1.2 and 7.0 ± 1.7 preoperatively to 12.7 ± 1.6 and 1.3 ± 1.2,respectively (P ＜ 0.05).Conclusions Angiography examination is routinely indicated for the patients with fracture-dislocation of the upper cervical spine perioperatively.Preoperative evaluation of the vertebral artery and its adjacent structures and effective intraoperative hemostasis can avoid uncontrollable bleeding during operation,reduce postoperative complications,and improve surgical outcomes.

Objective To explore the diagnosis and treatment of bronchogenic cyst in the abdomen.Methods Clinical data of 7 cases with abdominal bronchogenic cyst in the First Affiliated Hospital of China Medical University from October 2001 to February 2016 were retrospectively analyzed.Results Of the 7 patients,5 were male and 2 were female aging from 36 to 50 years with a median age of 37 years.Two cases were complaining for upper abdominal pain,5 cases were asymptomatic.Ultrasonography revealed hypoechoic or anechoic mass.Color doppler flow imaging showed no blood flow.Contrast-enhanced CT scans showed no obvious enhancement in 6 out of 7 cases,all patients underwent cystectomy successfully.Cysts were of unilocular in 6 cases and multilocular in one.Bronchogenic cysts were diagnosed by pathology in all 7 cases.Six patients were followed up from 1 month to 6 years with no recurrence.Conclusions Abdominal bronchogenic cyst is benign and postoperative prognosis is very good.

Objective The advantages of the treating allergic rhinitis (AR) by sphenopalatine ganglion stimulation with acupuncture were evaluated.Methods Databases including CBMDisk, CNKI, WanFang, VIP, Cochrane Library, PubMed, ProQuest, ChiCTR, ISRCTN, ClinicalTrials.gov and CENTRAL were searched from the beginning of database established to Jan 2015. All issues from Jan 2004 to Jan 2015 published onjournals Chinese Acupuncture & Moxibustion,Shanghai Journal of Acupuncture and Moxibustion, Acupuncture Research,Journal of Clinical Acupuncture and Moxibustion,Chinese Journal of Integrated Traditional and Western Medicine and Chinese Journal of Otorhinolaryngology in Integrative Medicine were searched by hand at meantime. All data were extracted based on the inclusive and exclusive criteria which was pre-designed, the Revman5.3 was applied for meta-analysis, and the studies qualities were analyzed by grade score.Results 118 articles were collected, 7 studies that involving 1 230 patients met the inclusive criteria. The result indicated that the sphenopalatine ganglion stimulation with acupuncture as the main treatment of AR showed the better total response rate compared to conventional drugs, theOR(95%CI) was 3.22(1.81 - 5.75); however the change of total symptom score had no statistical significant difference, the MD(95%CI) was 0.69 (-0.56 - 1.93), the change of IgE had no statistical significant difference,theSMD(95%CI) was -0.07 (-0.97 - 0.83).Conclusion The main treatment on AR by sphenopalatine ganglion stimulation with acupuncture may has better efficacy than western medicine. But due to the methodological biases existed in most studies, future high-quality RCTs were needed to be included into Meta-analysis to test today’s study conclusion.

OBJECTIVETo compare the effects of acupuncture on sphenopalatine ganglion and acupuncture on the common acupoints for life quality of patients with allergic rhinitis(AR).

METHODSEighty patients with AR,who were in accord with the inclusive criteria,were randomly divided into an observation group and a control group,40 cases in each one. Acupuncture on sphenopalatine ganglion was used in the observation group. The needle was inserted into the gap between zygoma and mandibular coronoid process about 55 mm. Acupuncture was adopted on the main acupoints,Yingxiang(LI 20),Yintang(GV 29) or Fengchi(GB 20) in the control group. The course was four weeks. Follow-up was applied one month after treatment. Rhinoconjunctivitis quality of life questionnaire (RQLQ),rhinitis symptoms scale and visual analogue scale(VAS) were evaluated at different time points before and after treatment,and follow-up was implemented to know the recurrence situation,satisfactory degree and adverse reaction.

RESULTS(1) RQLQ scores:along with treatment,the RQLQ scores were gradually apparently decreased in the two groups(<0.01),and the reducing trend from the first week to the second week of the observation group was more obvious than that of the control group. The differences of the RQLQ scores at all timepoints after treatment between the two groups were not statistically significant(all>0.05). The interaction of the time factor and the group factor had statistical significance(<0.01). (2) Rhinitis symptoms scores:along with treatment,the scores presented decreasing trend in the two groups(<0.01). The scores of the two groups after treatment and the interaction of the time factor and the group factor were not statistically different(both>0.05). (3)VAS scores:the VAS scores after treatment were lower than those before treatment(both<0.01). The differences before and after treatment were statistically significant(<0.01),with more change in the observation group(<0.05). (4)There was no statistical significance about the number of recurrence days between the two groups(>0.05). (5) Above 80 percent patients were content with the therapeutic method in each group,with no statistical difference(>0.05). (6)The method of the observation group spent less time. (7) Two patients with light adverse reaction came up in the observation group,but no special treatment was needed.

CONCLUSIONSAcupuncture on sphenopalatine ganglion acquires more obvious short-term effect than conventional acupuncture. It spends less time to relieve symptoms and improves life quality.

Objective To compare the difference of the sense of security and social support between left-behind women and non left-behind women in rural area.Methods Social Support Rating Scale (SSRS) and security questionnaire(SQ) were used to measure social support and sense of security of 98 left-behind women and 151 non left-behind women.The data was analyzed by SPSS17.0.Results ①In the social support rating,compared with the non left-behind women,the left-behind women has lower score in the total score((40.561±6.692) vs (59.722±8.699),t=18.530),determining the control factor((21.459±3.891) vs (30.013±4.950),t=14.450) and human security factor((19.102±3.737) vs (29.709±4.849),t=18.392) and the differences were statistical significant(all P＜0.05).②In the social support rating scale,left-behind women had lower scores in total score,exploitation degree of support,subjective support and objective support than the left-behind women(all P＜0.05).③The total score and each factor score of security scale,and the total score and each factor score of social support rating scale in the left-behind women showed significantly positively correlated(r=0.245-0.507,P＜0.05).Conclusion The sense of security and social support of the left behind women were worse than that of non left-behind women.It is necessary to carry out psychological intervention for them.

Background and purpose:The prognosis of pancreatic cancer is poor. This study aimed to evaluate the value of pancreatic CT perfusion on biological behavior assessment in pancreatic cancer. Methods:This study collected 78 cases of pancreatic cancer which diagnosed by the method of pancreatic CT perfusion, and detected the differences of the values of blood lfow (BF), blood volume (BV), permeability (per), peak value (PE) and time to peak (TTP) between normal pancreatic tissues and tumor tissues. Combined with clinical and pathological data. Besides, this study evaluated the relationship between perfusion parameters of tumor tissues and tumor size, lymph node metastasis, distant metastasis, preoperative serum CA199 level, Ki-67, p53, CEA, CA199, CD34 expression of tumor tissues. Results:The values of BF, BV, per and PE in pancreatic tumor tissue were signiifcantly lower than those in normal pancreatic tissue. The BF values of cases with high levels of serum CA199 and with CA199 positively expressed tissues were signiifcantly higher than those with negative expression. The PE values of cases with positive tissue expression of Ki-67 were significantly higher than those with negative expression. The TTP values of cases with positive tissue expression of CEA were signiifcantly lower than those with negative expression. The per values of well differentiated cases was signiifcantly higher than those of moderately/poorly differentiated cases. Conclusion:CT perfusion may have its value on assessment of tumor biological behavior in pancreatic cancer.

ObjectiveTo evaluate the efficacy of sphenopalatine ganglion stimulation with acupuncture for moderate-to-severe perennial allergic rhinitis.MethodsA total of 50 patients were recruited into a sphenopalatine ganglion stimulation group and a routine acupuncture group according to order of presentation, with 25 in each group. The sphenopalatine ganglion stimulation group received sphenopalatine ganglion stimulation with filiform needle, 1-2 sessions/week for 4 weeks. The routine acupuncture group received traditional acupuncture, withyingxiang(LI 20),yintang(GV29),fengchi(GB20),fengfu(GB16),zusanli(ST36) as the maln points, andyingxiang(LI 20),yintang(GV29),fengchi(GB20),fengfu(GB16),zusanli(ST36) as the adjunct points, 1-2 points from both the maln and adjunct points in each session, 2 sessions/week for 4 weeks. The nasal symptom score (2004 version), the total nasal symptom score (TNSS) and the total non-nasal symptom score (TNNSS) were used to evaluate symptom improvement. The Rhinoconjunctivitis Quality of Life Questionnalre (RQLQ) was used to assess the patients’ quality of life. The time to symptom alleviation, duration of symptom alleviation in every session and the recurrence duration during 1 month after the treatment were compared between the two groups.Results After the treatment, the score of the nasal symptom score (99.74 ± 31.89vs.196.83 ± 31.22;t=-4.912,P=0.001), TNSS (33.63 ± 12.37vs.71.82 ± 19.21;t=-3.463,P=0.003), TNNSS (33.63 ± 12.37vs.71.82 ± 19.21,t=-3.463,P=0.003) in the sphenopalatine ganglion stimulation were significant lower than those in the routine acupuncture group. Compared with the routine acupuncture group, the time to symptom alleviation was significant shorter (13.85 ± 4.21 minvs.45.63 ± 7.87 min;t=-1.763,P=0.008), while the duration of symptom alleviation was significant longer (37.92 ± 9.94 hvs.3.35 ± 1.23 h;t=7.637,P<0.01) after each session in the sphenopalatine ganglion stimulation group. Four weeks after the treatment, RQLQ score in the sphenopalatine ganglion stimulation group was significant lower than that in the routine acupuncture group (8.48 ± 3.71vs.37.68 ± 12.46;F=-7.312,P<0.01). The recurrence duration during 1 month after the treatment in the sphenopalatine ganglion stimulation group was significant longer than that in the routine acupuncture group (4.12 ± 2.15 dvs.23.53 ± 4.63 d;t=-8.879,P=0.003).ConclusionSphenopalatine Ganglion stimulation is superior to routine acupuncture in treatment of patients with moderate-to-severe perennial allergic rhinitis.

Objective:To explore the effect of Jinfeng particles on the expression of MMP-1/TIMP-1 in cartilage of gout arthritis rabbit′s articular,to explicit treatment mechanism of gouty arthritis. Methods:Perfusion of ethambutol and adenine,inject MSU into joint cavity of rabbits to copy the model of gouty arthritis. After successful mode,were randomly divided into model group,Jinfeng granules in high,medium and low dose group(19. 6,9. 8,4. 9 g/kg),allopurinol group(0. 028 g/kg),Tongfengding group(0. 448 g/kg) ,another blank group. Administered for 7 days. 2 h after the last administration,were taken and painted articular cartilage tissue,to observe the changes of the pathological morphology;and the use of immune cell detection of the expression of TIMP-1,MMP-1,MMP-1/TIMP-1 ratio calculation. Results: Jinfeng granule could obviously reduce the articular cartilage of rats with inflammatory cells infiltration,hyperemia, swelling and necrosis, could significantly inhibit the expression of MMP-1 ( P< 0. 05 ) , stimulate TIMP-1 activation(P< 0. 01),and significantly reduce the ratio of MMP-1/TIMP-1(P< 0. 05). Conclusion: Jinfeng particles can effectively inhibit the pathological damage of gouty arthritis,upside the expression of TIMP-1 and its mechanism may be related to the inhibition of MMP-1,regulated the balance of MMP-1/TIMP-1,and then exert anti-inflammatory effects.

Objective To detect the expression of MSI2 protein and mRNA in pancreatic ductal adenocarcinoma (PDAC) tissue,and investigate the correlation between the expression of MSI2 protein and the clinicopathological parameters.Methods The expression of MSI2 protein in 61 PDAC specimens and paired adjacent non-cancerous pancreatic tissues were detected by immunohistochemistry.Western blot and quantitative real-time PCR (QRT-PCR) were used to examine the expression of MSI2 protein and mRNA level in 10 PDAC specimens and adjacent non-cancerous pancreatic tissues.Then the relationship between MSI2 expression in cancerous tissues and clinicopathological parameters was analyzed.Results In 61 patients with PDAC,the expression rate of MSI2 protein was higher in cancerous tissues (63.9％) compared with that in paired non-cancerous pancreatic tissues (41.0％),and the difference between the two groups was statistically significant (t =2.809,P =0.007).The expression levels of MSI2 protein in 10 fresh PDAC specimens and adjacent non-cancerous pancreatic tissues were 0.748 ± 0.195 and 0.420 ± 0.171,and the expression level of MSI2 mRNA in PDAC specimens was as 2.507 ± 2.981 times as much of adjacent non-cancerous pancreatic tissues,and the difference between the two groups was statistically significant (t =3.689,P=0.005;t =2.660,P =0.026).The expression of MSI2 in cancerous tissues was only positively associated with the size of the tumor (x2 =5.096,P =0.024),but it was not associated with other parameters.The median survival of patients with high MSI2 expression was 321 d,and it was 730 d for patients with low MSI2 expression,and the median survival of patients with high MSI2 expression was significantly shorter than that of low MSI2 expression (x2 =6.706,P =0.010).Conclusions The expression MSI2 is up-regulated in PDAC and related to the tumor size.The patients with high expression of MSI2 protein have poor prognosis.