OBJECTIVE: To develop the single-tube one-step methylation variable position (MVP) analysis technology-single-tube post-digestion PCR-melting curve analysis (PDP-MCA). METHODS: Based on differentially methylated region (DMR) reported previously as the model, a set of primers with different melting temperatures of products in the two sides of MVP were designed. By using the FastDigest methylation-sensitive restriction enzyme (MSRE), DNA digestion, multiplex amplification, MCA detection and MCA profiles were performed in a single reaction tube. Same samples (peripheral venous blood, semen, and vaginal fluid, 5 samples each type) were tested by single-tube one step MVP and traditional MSRE-PCR MCA technology. To verify the feasibility of this method, the results were compared with that of the traditional technology. The MCA/HRM profiles of different samples were analyzed and compared. RESULTS: When the melting temperature of the fragments had a differential of 2 degrees C, the MCA melting peaks separated well, and MCA detection after multiplex amplification was successful. The single-tube PDP-MCA assay was developed, which integrated multiple reactions (digestion, amplification and detection) into one tube. By this method, the sample-specific profiles and data were analyzed in 2 h, which is similar to that of the traditional method. The rapid classifications of the samples were also realized. CONCLUSION Multiplex MVPs can be analyzed in a single closed-tube. The single-tube PDP-MCA technology is a simple, fast, and automatable method. It can be used for detection of DNA methylation variations.
DNA/isolation & purification* ; DNA Methylation/genetics* ; DNA Primers/genetics* ; Humans ; Multiplex Polymerase Chain Reaction/standards* ; Nucleic Acid Denaturation

Objective To observe the changes of plasma endothelin (ET), atrial natriuretic peptide (ANP) and hemodynamics during cardiopulmonary bypass (CPB) in patients undergoing cardiac valve replacement surgery. Methods Sixteen patients underwent cardiac valve replacement surgery were investigated. Plasma endothelin, atrial natriuretic peptide were measured with radioimmunoassay. Results Before the anesthesia induction, plasma endothelin levels [(2.68±1.8)] pg/ml were not significantly different from that in the control group [(2.22±0.7) pg/ml] while atrial natriuretic peptide levels was significantly higher [(738±559) pg/ml vs (72.7±14.5 pg/ml, P<0.001)]. Plasma ET concentration was constant during CPB period. Plasma ANP levels gradually decreased after CPB started and returned to basal levels when CPB stopped. During and after CPB, SVRI was significantly lower compared with that preanesthesia. SVRI was positively correlated with Hct. Conclusions Plasma ET concentration does not significantly vary during CPB period. The decrease of plasma ANP levels can attribute to hemodilution, which also accounts for lowed SVRI.

Objective To observe the changes of plasma endothelin (ET), atrial natriuretic peptide (ANP) and hemodynamics during cardiopulmonary bypass (CPB) in patients undergoing cardiac valve replacement surgery. Methods Sixteen patients underwent cardiac valve replacement surgery were investigated. Plasma endothelin, atrial natriuretic peptide were measured with radioimmunoassay. Results Before the anesthesia induction, plasma endothelin levels [(2.68±1.8)] pg/ml were not significantly different from that in the control group [(2.22±0.7) pg/ml] while atrial natriuretic peptide levels was significantly higher [(738±559) pg/ml vs (72.7±14.5 pg/ml, P<0.001)]. Plasma ET concentration was constant during CPB period. Plasma ANP levels gradually decreased after CPB started and returned to basal levels when CPB stopped. During and after CPB, SVRI was significantly lower compared with that preanesthesia. SVRI was positively correlated with Hct. Conclusions Plasma ET concentration does not significantly vary during CPB period. The decrease of plasma ANP levels can attribute to hemodilution, which also accounts for lowed SVRI.

Objective To analyze the relation between ~(99)Tc~m-DTPA renal dynamic imaging and pathological changes in patients with lupus nephritis (LN).Methods Ten normal control and 29 patients with LN underwent ~(99)Tc~m-DTPA renal dynamic imaging.The LN patients were divided into two groups:silent LN (SLN) group,18 patients;and obvious LN (OLN) group,11 patients.For each case,glomerular filtration rate (GFR),peak time (t_p),half excretion time (t_(1/2)) and the excretion rate at 20 min (R_(20)) were calculated.Assessment of renal function on the scintigraphic images was evaluated by nuclear medicine physicians.The t-test,Fisher'exact probability and R×C association were used for data analysis.Results There were significant differences between normal people and two goups of LN in tp(t=5.3,9.3,both P＜0.05),t_(1/2)(t=6.9,12.0,both P＜0.05)and R_(20)(t=10.1,12.1,both P＜0.05).As to GFR,there was significant decrease in OLN patients(t=4.1,P＜0.05),but not in SLN patients(t=1.7,P＞0.05).Diagnoses of renal function by renal dynamic imaging were compared with the renal pathological changes (r=0.2273,P＜0.05).Conclusions ~(99)Tc~m-DTPA renal dynamic imaging is useful for evaluation of the early stage renal function for LN patients and to diagnose LN patients with no symptom of renal impairment.It may help to assess the degree of renal parenchymal damage while obviating the need for renal biopsy in these patients.

Apyrase activities in some tissues and cells, such as peripheral vascular endothelial cells, have been reported, but these in endocardium endothelial cells have not been reported. The present study was to characterise the properties of bovine endocardium endothelial cells (BEEC)-associated apyrase. Apyrase activity was assayed by inorganic phosphate release, which could be inhibited concentration-dependently by NaN3, an apyrase inhibitor. NaF (20 mmol/L), another inhibitor of apyrase, also markedly inhibited the activity. EDTA or EGTA (1 mmol/L) could also inhibit the activity completely. However, the inhibitor for Na+/K+-ATPase, ouabain (3 mmol/L) did not affect the enzyme activity. BEEC apyrase activity was dependent on divalent cations (Ca2+ or Mg2+) and pH value.

OBJECTIVETo carry out expert survey for traditional Chinese medicine (TCM) syndrome characteristics of different clinical types of coronary artery disease (CAD).

METHODSBy using Delphi method, we carried out two rounds of nationwide expert surveys for modern TCM characteristics of syndrome elements and syndrome types of CAD.

RESULTSBased on expert consensus, qi deficiency, blood stasis, phlegm turbidity, qi deficiency blood stasis, and intermingled phlegm and blood stasis are common TCM syndromes for different clinical types of CAD. Of them, qi stagnation, blood stasis, phlegm turbidity, heat accumulation, cold coagulation, yang deficiency, deficiency of both qi and yang were more often seen in patients with unstable angina than in those with stable angina. Qi deficiency, yin deficiency, and deficiency of both qi and yin were less seen. We could see more excess syndrome and less deficiency syndrome (such as qi deficiency, yin deficiency, etc.) in acute ST-segment elevation myocardial infarction (STEMI) than acute non-ST-segment elevation myocardial infarction (NSTEMI). Qi deficiency, blood stasis, water retention, yang deficiency, phlegm turbidity, yin deficiency, Xin-qi deficiency, and qi deficiency blood stasis induced water retention are the most common TCM syndrome types of CAD heart failure (HF). Blood deficiency, yin deficiency, heat accumulation, deficiency of both Xin and Pi, deficiency of both qi and blood, deficiency of both qi and yin, yin deficiency and fire hyperactivity were more often seen in CAD arrhythmias.

CONCLUSIONSTCM syndrome distributions of different clinical types of CAD have common laws and individual characteristics. Results based on the expert consensus supplied evidence and support for clinical diagnosis and treatment of CAD.

Angina Pectoris ; Angina, Unstable ; China ; Coronary Artery Disease ; diagnosis ; therapy ; Coronary Disease ; diagnosis ; Data Collection ; Heart Failure ; diagnosis ; Humans ; Medicine, Chinese Traditional ; methods ; Qi ; Syndrome ; Yang Deficiency ; diagnosis ; Yin Deficiency ; diagnosis

OBJECTIVETo study the bone state with ultimate stress by examining biomechanical distribution of upper femur in Chinese, in order to accumulate more experiences for clinical work.

METHODSTotally 60 Chinese femurs from fresh cadavers were randomly divided into two groups. All the femurs were cleaned, and the body age ranged from 36 to 72 years old, averaged 56.4 years, including 41 from males, and 19 from females. These two groups underwent mechanical stress and bending stress tests. Special mechanical laboratory and machines were used to get the information. Results about the loading value at each testing point under stress were collected.

RESULTSThe four faces of the upper femur suffered different stress under external forces. The bone on upper femur can tolerate more mechanical stress than bending stress. Medial and lateral region of the femur neck and the rear side of the small tuberosity section were themain position enduring the vertical stress. The rear position of the base femur neck and the small tuberosity section were the main regions enduring the bending stress. Those main positions had strong cancellous bones. The intertrochanteric fracture fixation and artificial femoral stems were designed depending on this biomechanical basis.

CONCLUSIONAccording to our experiment result, doctors need to chose more effective fixations for upper femur fracture, and femoral stems for the patients. More information should be collected by further researches.

Adult ; Aged ; Biomechanical Phenomena ; Female ; Femur ; chemistry ; Humans ; Male ; Middle Aged ; Stress, Mechanical

Objective To examine HIV prevalence and related risk factors among men who have sex with men(MSM) in Chongqing, and to explore the feasibility of using respondent-driven sampling (RDS) in the survey. Methods Based on results from formative research, a RDS survey was designed and conducted to collect demographic, behavioral and serologic data. RDSAT was used to calculate point estimation and confidence intervals. SPSS was used for bi-variate analysis using RDSAT exported weighed data. NETDRAW was used to draw network diagram. Results Among 617 subjects recruited, the adjusted HIV and syphilis prevalence rates were 16.8% and 10.9%, respectively. 73.0% of the subjects were 20 to 29 years old and 72.9% were officially registered residents of Chongqing. 83.4% were single, with the proportion of students the highest, accounting for 24.6%. During the last six months, 83.4% of them reported ever having anal sex, and 54.0% reported having unprotected anal sex. Conclusion This survey confirmed that Chongqing had a higher reported HIV prevalence among MSM than from other Chinese cities. Comprehensive intervention services were required to address this alarmingly high prevalence, with focus on intervention through internet and those having syphilis infection. RDS seemed one of the effective ways of recruiting hidden MSM populations in Chongqing which had a large population of active MSM who did not frequently visit MSM venues as compared with snowball method.

0.05);2)After 12,24,36 months' treatment,BP was decreased significantly in each group (P0.05).Conclusion Both combined spirono- lactone/HCTZ and captopril/HCTZ significantly reduced BP and LVMI or LVMI and the maguitude of reduction was further enhanced after prolonged treatment.

OBJECTIVEAccording to the information obtained from the revision of total hip arthroplasty, the situations of the artificial femoral stem under physiological stress were analyzed preliminarily. In order to accumulate clinical experience, the discussions about how to enlongate the life of artifical joints were performed.

METHODSFifty-three patients required revision operations were selected, including 28 males and 25 females,with an average age of 74.66 years old (61 to 84 years old). The average life of artificial joints was 18.24 years (3 to 27 years). The initial weightbearing was started 11 days (5 to 16 days) after the first operation. The main reasons for the revision were the spontaneous loosening and trauma. Based on imaging and surgical findings, as well as the histological pathology, the situations of the stems in human bodies were determined.

RESULTSFemoral prosthesis would sink under physiological stress. The design of femoral stems, motion of patients', and bone growth determined the secondary stability. Straight stems were helpful for physiological subsidence of prosthesis, achieving the stability with close bone-implant contact by bone growth,which allowed the patient to do early landing exercise. The collared prosthesis and prosthesis combined with internal fixation limited the subsidence of femoral stems. Their stability depends on the time of initial placement and the bone growth for supporting. Delaying the inital weightbearing was suggested for patients with these protheses.

CONCLUSIONAccording to the actual situation of the patients, the appropriate chosen of femoral stems and time to perform the weightbearing can extend the life of the femoral prosthesis.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; adverse effects ; methods ; Female ; Hip Prosthesis ; Humans ; Male ; Middle Aged ; Prosthesis Design