Objective: To explore the differential expression profiles of DNA methylation sites/regions and potential molecular mechanisms in the peripheral blood of coronary heart disease (CHD)-induced unstable angina pectoris patients with or without Qi deficiency and blood stasis syndrome, and to provide scientific evidence for the conbination of disease and syndrome. Methods: According to the pre-determined inclusion and exclusion criteria, the study subjects were enrolled and divided into two groups namely CHD-induced unstable angina group (G group) and healthy control group (J group) to conduct “disease” analysis, while G group was further divided into Qi deficiency and blood stasis syndrome group (case group) and non-Qi deficiency blood stasis syndrome group (control group) to perform “syndrome” analysis. The general data and clinical information of the study subjects were collected. The peripheral venous blood was extracted on an empty stomach, and the Illumina Infinium MethylationEPIC BeadChip (850K methylation chip) was used to detect the differential expressionprofiles of DNA methylation in each group, ChAMP software (V 2.14.0) was used for the differential methylation data analysis, with a threshold of the adjusted P value (adj.P.val) < 0.01. Gene Ontology (GO) and Kyoto Encyclopedia of Genomes (KEGG) were employed for the functional and pathway enrichment analyses of related mapped genes. Results: A total of 263 differentially methylated CpG positions (DMPs) were screened out between G and J groups, including 191 hypermethylated positions such as cg05845204 and cg08906898, and 72 hypomethylated positions such as cg26919182 and cg13149459. These positions were mainly mapped to 148 genes encompassing RNA binding motif protein 39 (RBM39), acetyl-CoA acyltransferase 2 (ACAA2), protein phosphatase 1 regulatory subunit 12B (PPP1R12B), and the dual-specificity tyrosine phosphorylation-regulated kinase 2 (DYRK2). GO functional enrichment analysis revealed that the genes of the DMPs were primarily enriched in protein localization to chromosomes, regulation of cell morphogenesis, negative regulation of calcium-mediated signals, etc. KEGG pathway analysis suggested that the genes were mainly enriched in fatty acid metabolism and endocytosis pathways. In addition, a total of 23 differential methylation regions (DMRs) were identified, with overlapping genes such as transmembrane protein 232 (TMEM232), ribosomal protein large P1 (RPLP1), peroxisomal biogenesis factor 10 (PEX10), and forkhead box N3 (FOXN3) recognized. It was found that GO functions were mainly enriched in the negative regulation of Ras protein signal transduction, small GTPase-mediated signal transduction, negative regulation, etc. A total of 1 703 differential methylation sites were screened out between case and control groups, including 444 increased methylation positions such as cg05573767 and 1 259 decreased methylationpositions such as cg19938535, and cg03893872. These positions were mapped to 1 108 genes such as ribosomal protein S6 kinase A2 (RPS6KA2), leucine rich repeat containing 16A (LRRC16A), and hedgehog acyltransferase (HHAT). According to the GO functional enrichment analysis, the genes relating to the DMPs were mainly enriched in biological functions such as transmembrane receptor protein serine/threonine kinase signaling pathway and axonogenesis. The KEGG pathway enrichment analysis suggested the involvement of Rap1 signaling pathway, adenosine 5’-monophosphate-activated protein kinase (AMPK) signaling pathway, etc. A total of 21 DMRs were identified, including 22 overlapping genes such as mucin 4 (MUC4), three prime repair exonuclease 1 (TREX1), and LIM homeobox 6 (LHX6). GO analysis demonstrated that the genes primarily participated in molecular functions such as positive regulation of transmembrane transport, regulation of fatty acid metabolism, and copper ion binding. Conclusion This study reveals the methylation patterns of DMPs and DMRs in patients with Qi deficiency and blood stasis syndrome caused by CHD-induced unstable angina pectoris. Potential epigenetic regulation of fatty acid metabolism, Rap1 signaling, and other molecular functions are involved in the development of CHD between the "disease" and "syndrome".

Objective ToinvestigatethefeasibilityofpredictingtheKi-67expressionusing MRandclinicalfeaturesofbreast cancer.Methods 52patientswith56invasivebreastcarcinomawereretrospectivelyinvestigated.Eachsubjectunderwentpresurgical breastMRI,whichincludeddynamiccontrastenhancementandDWI(b=0s/mm2,1000s/mm2).Clinicalcharacteristicsand MR findingswerecollected,includingage,ER ,PR ,HER2,tumorsize,location,number,spiculesign,lobulationsign,margin,TICandADCvalues. Ki-67expressionwasrecordedbyimmunohistochemicalstaining,whichwasdividedintohighexpression(≥20%)andlowexpression (<20%).Leastabsoluteshrinkageandselectionoperator(Lasso)wereperformedtoselectthefeaturesmostassociatedwithKi-67 expression.ThenomogramwasconstructedtopredictKi-67expression,aswellasAUCand95%confidenceinterval(CI)werecalculated.Results TICtypesandADCvaluesassociatedwithKi-67expressionwereidentifiedbyLassoregression.Thenomogram predictedthehigh andlow Ki-67expressionwithanAUCof0.827 (95% CI:0.713-0.940).TheKi-67expressionwasnegativelycorrelatedwiththe ADCvalues(r=-0.430,P=0.003).Ki-67expressionwassignificantlycorrelatedwithTICtype,andTICⅢwasmorelikelytohavehigher expressionofKi-67.Conclusion ThereisindividualandrespectivecorrelationbetweentheexpressionlevelofKi-67andTICtype, ADCvalue.TheexpressionlevelofKi-67canbepredictedtosomeextentbyADCvaluesandTICtypes.

Objective To cxplore the optimal levels of serum calcium,phosphorus and intact parathyroid hormone (iPTH) in peritoneal dialysis (PD) patients.Methods This study is a single center,retrospective cohort study.The associations between serum calcium,phosphorus and iPTH and all-cause mortality in 217 PD patients were analyzed.All patients started PD between January 1,2008 and April 30,2016 were enrolled and followed up to December 31,2016.At baseline and every 3 months,biochemical and therapeutic information was collected.Cox proportional hazard regression models and cubic splines analysis were employed to assess the lowest mortality risk ranges in serum markers of bone metabolism.Results There was no significantly difference between patients within target ranges based on KDOQI or KDIGO guideline and those outside the target ranges by Kaplan-Meier survival analysis.The lowest mortality risk ranges were 2.17-2.40 mmol/L for serum calcium,1.20-1.67 mmol/L for serum phosphorus and 180-350 ng/L for serum iPTH by using Cox models and cubic splines analysis.Moreover,cumulate survival had significant difference between patients within the descriptive ranges and those out of the descriptive ranges at time-averaged values but not at baseline values.Conclusions The optimal time-averaged ranges of PD patients are 2.17-2.40mmol/L for serum calcium,1.20-1.67 mmol/L for serum phosphorus and 180-350 ng/L for serum iPTH.These ranges need further validation by large population studies to further conform.

Objective This study investigates the relationship of serum irfflammatory cytokine levels with intracranial pressure (ICP) in patients with severe traumatic brain injury (TBI) after surgery.Methods A total of 32 cases with severe TBI and placement of ICP monitor were prospectively enrolled.Serum was collected before surgery and every 12 h after surgery.Cytokines levels of interleukin (IL)-1β,IL-8,and tumor necrosis factor (TNF)-α were analyzed and compared with outcome of patients.Hourly values of ICP were recorded.The degree of ICP above treatment threshold (20 mm Hg) were calculated every 12 h as pressure times time dose(PTD-ICP20),which was compared with serum cytokine levels before (Pre) and after (Post) the 12-hour time period using linear regression method.Results Serum IL-1β (P＜0.05),IL-8 (P＜0.01) and TNF-a (P＜0.01) levels elevated dramatically after severe TBI and were significantly associated with outcome of patients.Mean PTD-ICP20 was (42.9 ± 60.2)mm Hg/h and was correlated with increased Pre-IL-8 (r=0.554,P＜0.001),Pre-TNF-α (r=0.597,P＜0.001),Post-IL-8 (r=0.629,P＜0.001) and Post-TNF-α (r=0.538,P＜0.001) levels.Conclusion Serum IL-8 and TNF-α demonstrated the most promising candidate bio.rnarkers of impending ICP elevation in this study.These findings indicate a feasible way of monitoring patients with severe TBI.

Objective To determine the effect of omental flap wrapping of pancreaticoduodenectomy on postoperative pancreatic fistula and intra-abdominal bleeding in patients undergoing pancreaticiduodenectomy through meta-analysis of published studies.Methods Articles about whether omental flap wrapping could prevent postoperative pancreatic fistula and intra-abdominal bleeding were searched in databases of MEDLINE,PUBMED and EMBASE.14 English article were found and according to including and excluding criteria,4 eligible articles with a total of 2971 patients were selected.Random effects model was used to analyze odds ratios and 95％ confidence intervals (CIs).Results 1129 patients were in omental wrapping group,and 1842 patients in nonomental wrapping group.Omental roll-up during pancreaticoduodenectomy could not prevent postoperative pancreatic fistula (OR =0.81,95 ％ CI 0.40～1.63,P＞0.05) or postoperative abdominal bleeding (OR=0.67,95％ CI 0.28～1.59,P＞0.05).The sensitivity analysis showed the pancreatic fistula rate was lower in the nonomental roll-up group than that in the omental roll-up group (OR=1.24,95％ CI 1.03～1.50,P＜0.05).Conclusions Omental wrapping can not decrease the risk of pancreatic fistula and abdominal bleeding after pancreaticoduodenectomy.Further randomized controlled trials are needed to identify the effect of omental wrapping technique for pancreaticoduodenectomy.

Objective Toinvestigatetheeffectofwhetherposteriorcommunicatingartery(PCoA) patent or not on hemodynamics of extracranial internal carotid artery (EICA)in patients with moyamoya disease.Methods Atotalof103consecutivepatientswithmoyamoyadisease(unilaterallesion) detected by transcranial color Doppler ultrasonography (TCD)and Color Doppler flow imagining (CDFI), and confirmed by digital subtraction angiography (DSA)from August 2008 to April 2014 were enrolled. According to the patency of PCoA showed by DSA,the patients were divided into either a PCoA non-patent group (n=83)and a PCoA patent group (n=20). Eighty healthy subjects screened by TCD and CDFI in the same period were used as a control group. CDFI and TCD were used to detect the peak systolic velocity (PSV),end diastolic velocity (EDV),vascular resistance index (RI)and/or pulsatility index (PI)of EICA,and posterior cerebral artery (PCA)in subjects of both groups. Results (1 )There were significant differences in PSV,EDV and RI among the three groups (F=5. 76,12. 24,and 8. 10,respectively;all P<0. 05). The PSV of vascular EICA (42 ± 15 cm/s)and EDV (14 ± 7 cm/s)on the lesion sites in patients of the PCoA non-patent group were all lower than those in the control group (92 ± 24 and 38 ± 11 cm/s). The differences were statistically significant (all P<0. 05). The PSV of vascular EICA on the lesion sites in patients of the PCoA patent group was lower than that of the control group (P<0. 05),but EDV was higher than the control group (P<0. 05). (2)Compared with the control group,the RI value was increased in the PCoA non-patent group (P<0. 01)and that was decreased in the PCoA patent group (P<0. 01). (3)PSV and EDV of PCA in the moyamoya disease group were higher than those of the control group (P<0.05),andthePIvaluewaslowerthanthatofthecontrolgroup(P<0.01).Conclusion Whether PCoA patent or not in patients with moyamoya disease have different influences on hemodynamic parameters of EICA. PSV and EDV of the patients with PCoA non-patent are lower than those of the healthy subjects and their RI values are increased. EICA of the patent group is lower than PSV and RI of the control group, and their EDV is increased.

The amino group PEGylation of rhIFNomega with monomethoxy polyethylene glycol succinimidyl succinate (mPEG-SS, 20 000) was investigated, and the modified mixture was separated and purified by ion exchange chromatography and gel filtration chromatography. Under the optimized purification conditions, the average content ofmono PEG-rhIFNomega in the collect liquid reached 182 microg x mL(-1). The average purified yield of mono PEG-rhIFNomega exceed to 22%, and the purity of mono PEG-rhIFNomega was greater than 98% by SDS-PAGE and RP-HPLC. Relative molecular mass of mono PEG-rhIFNomega was 43 790 detected by MALDI-TOF MS. The apparent molecular mass measured by SDS-PAGE was about 60 810. The purified PEG-rhIFNomega has the characteristics of typical PEGylated protein. Activity reservation rate of mono PEG-rhIFNomega was 15.0%, while the antigenicity decreased by at least 64 folds. In addition, the acid stability, thermal stability and stability in serum and trypsin solution of mono PEG-rhIFNomega were markedly better than those of the rhIFNomega. The pharmacological properties of mono PEG-rhIFNomega were significantly improved. The prepared PEG-rhIFNomega might be developed to a novel safe and long-acting interferon.

ObjectiveTo evaluate the performance of a three-gene typing system in the determination of Treponema pallidum (Tp) genotypes.MethodsTo determine the genotypes of Tp,three targets were assessed,including the number of 60 base-pair repeats,restriction fragment length polymorphism(RFLP) pattem of tprEGJ gene after MseI digestion and the sequence of tp0548 gene.The DNA extracted from the Nichols strain of Tp served as the positive control,and that from the moist ulcer of patients with genital herpes and negative RPR or TPPA test results served as the negative control.To validate the typing method,clinical specimens were collected from the moist skin lesions of patients with primary or secondary syphilis,and subjected to the amplification of polA gene by PCR.The enhanced molecular typing system was used to determine the genotypes of Tp in Tp DNA-positive specimens.ResultsThe Nichols strain harbored a genotype of 14a/a.No amplification of any of the three target genes was found in the negative control.The arp gene,tprEGJ gene and tp0548 gene were amplified from 94.1％,91.2％ and 94.1％ of the 40 clinical specimens,and the genotype was successfully determined by the three-gene typing system for 91.2％ of the clinical Tp strains.The predominant type of arp,tprEGJ and tp0548 genes was 14 repeats,d and f,respectively in these clinical Tp isolates.ConclusionThe enhanced molecular tying method for Tp exhibits high sensitivity,specificity and discrimination potential.

Objective To investigate the effect and the significance of soft tissue repairing and functional reconstruction in limb salvage operation on extremity malignant bone tumors by individulized prosthesis replacement after malignant tumor resection with the help of Neo-adjuvant chemo-therapy. Methods A total of 78 patients with malignant bone tumor were recruited, including 42 males and 36 females. Aged 19～61, with an average of (29.12±9.47).Tumor in 14 patients was in the proximal humerus, 11 in the proximal femur, 27 in the distal femur, 3 in femoral middle part, and 23 in the proximal tibial. There were 29 cases of osteogenic sarcoma, 18 chondroma sarcomatosum, 7 maligant enchondroma with pathological fracture, 20 maligant giant cell tumor,and 4 maligant inflammatory myofibroblastoma of the bone. Soft tissue repairing and functional reconstruction were carried out together with individualized prosthesis replacement. The type of the prostheses was as follows: 14 patients had long stem humerus head prosthesis, 50 made hinged knee prostheses with femoral or tibial component, 3 whole femur replacements, 7 long stem femoral head prostheses, 4 long stem hip prostheses.Results All patients were followed up for an average of (26.80±8.06) months (4～37 months) and postoperative functions were estimated according to Enneking system. Among the 78 patients, results in 48 (61.5%) were excellent, 17(21.8%) were good,10 (12.8%) were fair,and 3(4.9%) were poor. The satisfactory rate was 83.3%. Conclusion Soft tissue repairing and functional reconstruction in limb salvage operation on extremity bone malignant tumors by individualized prosthesis replacement not only spare the limbs, but also keep their function.It can remove the psychologic obstacles caused by extremity absence, and is effective for bone malignant tumor.

Objective To study the clinical value of vascular ultrasound examination in detection of carotid artery disease. Methods 53 cases of ischemic cerebrovascular disease who served as experimental group,tested 387 vessels; and used the same period 55 cases of healthy persons as control group, tested 394 blood vessels. Comparison of two blood flow parameters between groups with differences in plaque detection. Results The results of vascular ultrasound examination could detect the visual display and quantitative carotid artery lesions in the experimental group a significant thickening of carotid artery IMT,carotid artery plaque prevalence was significantly higher than control group (P ＜0.01). Conclusion Vascular ultrasound examination was an effective screening method to detect ischemic cerebrovascular disease.