Objective To investigate the feasibility of Chelex DNA extraction from thin blood smears for genetic analysis, and to develop smear-based nested polymerase chain reaction (PCR) of the 18S RNA of Plasrnodium falciparum. Methods Chelex-100 which was chelating ion exchange resin was used to extract DNA from Giemsa-stained or unstained thin blood smears of different concentrations of Plasmodium falciparum. With the extracted DNA as the template, 18S RNA gene was amplified by nested PCR to test the susceptibility of Chelex method. Results Positive band of 205bp appeared in nested PCR with DNA extracted from Giemsa-stained or unstained thin blood smears of patient with falciparum malaria. Using the Chelex method, the detection limits of the smear-based nested PCR were 1.5 × 101 parasite/μL blood for Giemsa-stained and 1.5×10-1 parasite/μL blood for unstained thin blood smears. Conclusions Chelex DNA extraction is a simple and efficient method for extracting trace amount of DNA from thin blood smear. The smear-based nested PCR developed in this study is feasible to identify the gene from reserved thin blood smears and will provide a new approach for clinical diagnosis and study of molecular epidemiology.

Objective To investigate the effects of valsartan on platelet and endothelial function in atrial fibrillation dogs induced by chronic rapid atrial pacing.Methods From Sep.2001 to Jun.2003,14 dogs paced at 400 bpm in the right atrium for 6 weeks were randomly divided into control group and valsartan group.The dogs in valsartan group received valsartan(30 mg?kg~ -1 ?d~ -1 )1 week before rapid atrial pacing until pacing stop.The plasma von Willebrand factor(vWF),endothelin-1(ET-1),angiotensin Ⅱ(AngⅡ)levels and blood platelet aggregation rate were measured before and after rapid atrial pacing.Results (1)Plasma AngⅡ level elevated significantly in both groups,indicating activation of renin-angiotensin system.(2)The maximam platelet aggregation rates by four different agonists(Adenosine diphosphate,Epinephrine,Collagen,Arachidonic acid)increased significantly in control group dogs.To some extent,valsartan could inhibit platelet activation caused by chronic rapid atrial pacing.(3)There were significant elevations in the plasma levels of vWF and ET-1 in control group dogs.Valsartan could meliorate endothelial function and decrease the concentrations of vWF and ET-1.Conclusion Valsartan can inhibit platelet activation and meliorate endothelial function in chronic rapid atrial pacing dogs.

Linolenic acid containing selenium was synthesized and brought into emul- sion(82). Its aatitumor effects were tested in vitro and in vivo. As a result, when 82 was injected peritoneally into mice(200mg?kg-1?d-1), its inhibitory rate on mouse S180 was 42.6%(P

70%, P143%, p

ObjectiveTo explore the pathological effect of Anti-stenosis No.1 which had effect of vivid Qi and promoting blood flow,eliminate sputum and circulating collaterals on restenosis after carotid artery angioplasty in experimental rats.Methods48 rats were randomly divided into the blank group,the hyperlipid group,the small-dose group and the large-dose group.Restenosis model in carotid artery after angioplasty was built by high-dose cholesterol diet and three times balloon injury.Chinese herbs were administered for three months.Lumen area,intimal area,media area and intimal area/media area in the restenosis region were calculated.ResultsLumen area and intimal area were very more significantly changed in the hyperlipid group,the small-dose group,and the large-dose group compared to the blank group(P<0.01).Media area and intimal area/media area were very more significantly changed in the hyperlipid group and the small-dose group compared to the blank group(P<0.01).There was no significant difference in the large-dose group.Compared to hyperlipid group,lumen area significantly increased in the large-dose group(P<0.01)),intimal area significantly decreased in the small-and large-dose groups(P<0.05),and media area significantly decreased in the large-dose group(P<0.01).There was a significant difference in media area between the small-and large-dose groups(P<0.05).ConclusionAnti-stenosis No.1 can significantly inhibit restenosis by inhibiting intimal and media hyperplasty,and the effect is dose dependent.

AIM To observe whether acute stimulation of BRL 37344, a β3-adrenergic receptor (β3-AR) agonist, has the same effects of exacerbating hemodynamics and increasing in β3-AR expression on rats with failing heart as chronic administration. METHODSRats received two doses of isoprenaline (Iso, 340 mg·kg-1, sc, with a 24-h interval) to prepare heart failure model. After 8 weeks, rats were given iv BRL 37344 0.4 nmol·kg-1·min-1 for 10 min. Hemodynamics were measured at 0, 10, 30 min, 1, 2, 3 h, 1, 2 and 7 d after BRL 37344. Levels of β-AR mRNA in myocardium were measured at 0, 1, 2 and 7 d after BRL 37344 by reverse transcription-polymerase chain reaction. RESULTSCompared with Iso group, heart rate, left ventricular end systolic pressure and +dp/dtmax were significantly higher, and left ventricular end diastolic pressure was significantly lower during 1-3 h after BRL 37344 injection in Iso+BRL group. Then, they were restored to the same level as that prior to BRL 37344 injection. Compared with normal control, the levels of β1-, β2- and β3-AR mRNA displayed no significant change in BRL group;the level of β1-AR mRNA was lower and the level of β3-AR mRNA was higher in Iso group. In Iso+BRL group, much more lower β1-AR mRNA level and much higher β3-AR mRNA level were shown on d 2 and d 7 than Iso group. CONCLUSION Acute administration of β3-AR agonist has a shorter improved hemodynamics. But it caused the same result as chronic administration in reduction of β1-AR mRNA and increment of β3-AR mRNA in failure hearts, which may aggravate the cardiac functions.

Objective To investigate and analyze the mineral and bone disorder (MBD) in the patients with chronic kidney disease (CKD),reveal the change of related indexes of CKD-MBD.Methods A cross-sectional study was carried out in the First Affiliated Hospital of Harbin Medical University.From October 2011 to May 2014,1318 inpatients and hemodialysis outpatients were enrolled.Parameters related to MBD,including serum phosphorus (P),total calcium (t-Ca),intact parathyroid hormone (iPTH) and alkaline phosphatase (AKP) were analyzed.Last,it was analyzed with multiple regression analysis to related factors of the secondary hyperparathyroidism (SHPT) in patients with CKD.Results Serum calcium,phosphorus and iPTH had no obvious abnormalities at the early stages of CKD [GFR ＞ 60 ml· min-1· (1.73 m2)-1],and relatively stable before GFR ＞ 30 ml· min-1· (1.73m2)-1.After entering the CKD4 stage,serum phosphorus,iPTH increased sharply and serum calcium decreased obviously along with the decreased glomerular filtration rate (GFR).Serum P,t-Ca and iPTH levels were statistically significant in CKD 1 to 5D patients,respectively,serum P:(1.13±0.20) mmol/L,(1.14±0.22) mmol/L,(1.26±0.23) mmol/L,(1.48±0.34) mmol/L,(2.05±0.61) mmol/L and (2.08±0.58)mmol/L;serum t-Ca (mmol/L) (2.35±0.13) mmol/L,(2.35±0.12) mmol/L,(2.35±0.15) mmol/L,(2.26± 0.18) mmol/L,(2.07±0.29) mmol/L and (2.31±0.26) mmol/L;iPTH:57.8(45.6,91.8) ng/L,54.1(37.8,74.6) ng/L,71.6(45.8,102.2) ng/L,131.1(81.7,205.1) ng/L,277.5(173.6,395.3) ng/L and 354.9 (194.4,720.3) ng/L;The stepwise logistic regression analysis showed:hypocalcemia (OR=3.32,P ＜ 0.01) and decreased GFR (OR=5.28,P ＜ 0.01) were independent risk factors of iPTH elevation at stage CKD3～ 5.Conclusions From the beginning of the CKD3 stage,serum t-Ca,P,iPTH level began to be relatively abnormal as renal function declined.Hyperphosphatemia,SHPT has not been improved significantly in CKD5D stage patients even with hemodialysis.The regulation of hemodialysis on serum calcium showed overcorrecting phenomenon.

Objective To investigate the potential role of soluble triggering receptor expressed on ayeloid cells-1(sTREM-1) expression in serum,endotracheal aspiration (ETA),bronchoalveolar lavage fluid (BALF) and exhaled breath condensate (EBC) as early biomarkers for the diagnosis of ventilator-associated pneumonia (VAP) in patients with acute ischemic stroke.Methods One hundred and thirty-two patients with clinically suspected VAP were prospectively included in this multicenter study.The levels of sTREM-1 in serum,ETA,BALF and EBC were analyzed for diagnostic evaluation at the time of VAP clinically suspected.The bacterial count over 104/CFU as a gold standard for VAP,and the receiver operating characteristic curves were used to identify the ideal cutoff values.Results VAP was confirmed in 76 patients (57.58％).In VAP patients (VAP group) and non-VAP patients (non-VAP group),the level of sTREM-1 in BALF was 32.35 (30.08-41.72) and 18.92(11.89-31.72) ng/L,and the level of sTREM-1 in EBC was 1.57 (1.02-2.61) and 0.41(0.19-1.61)ng/L respectively.The level of sTREM-1 in BALF and in EBC in VAP group was significantly higher than that in non-VAP group (P ＜0.05).The optimum cutoff value for sTREM-1 in BALF according to the maximum Youden index was 23.61 ng/L.This cutoff value had 85.5％ sensitivity and 73.1％ specificity,with 0.813 area under the curve.sTREM-1 in BALF had excellent correlation with that in EBC (R2 =0.78,P ＜ 0.05).Conclusions The results of this prospective study suggest that sTREM-1 levels in BALF and EBC have better roles in facilitating the diagnosis of VAP and thus may be practically recommended to guide the administration of antibiotics when VAP is suspected.

Objective To evaluate the feasibility and value of quantitatively assessing left ventricular systolic function in rats with myocardial infarction (MI) by myocardial contrast echocardiography (MCE) based on mathematical morphology algorithm.Methods SD rats model of MI were mede,then MCE were performed before MI and at 1,3 weeks after MI.In vitro study:based on the principle of binary morphology,segmentation of endocardium was acquired using dilation,erosion,closed operations and connected domain object tag-based.The automatic segmentation of endocardial contour was compared with the manual segmentation boundary.In vivo study:the left ventricular area was calculated using the software,and the area variation fraction (AVF),a new index of left ventricular systolic function,were acquired.The correlation between AVF and left ventricular ejection fraction (LVEF) was analyzed.The value of AVF for diagnosis for left ventricular dysfunction was evaluated by ROC curve.Results ① The cross-correlation coefficient (CCC) and percent error (PE) between automatic contours and manual boundary were more than 0.90 and less than 9％,respectively.② AVF correlated positively with LVEF (r =0.934,P ＜ 0.001).③ ROC analyses showed the area under curves for AVF diagnosing left ventricular dysfunction was 0.834.The best cutoff value was 27.5 ％ for diagnosing left ventricular dysfunction with the sensitivity of 85 ％ and specificity of 60 ％.Conclusions Left ventricular endocardium can be identificated automatically and rapidly by MCE based on mathematical morphology algorithm.AVF can assessed quantitatively left ventricular systolic function in rats with myocardial infarction.

Objective To find out the distribution of TCM constitution on the patients with stroke in Beijing. Method The TCM Constitution Questionnaire was used to evaluate 489 patients with stroke in three hospitals in Beijing. Results The qi-deficiency constitution (140 patients, 28.63%), yin-deficiency constitution (95 patients, 19.43%), blood-stasis constitution(89 patients, 18.20%) and phlegm-heat constitution (73 patients, 14.93%) were the most common constitution, while qi-depression constitution (22 patients, 4.50%) and yang-deficiency constitution (20 patients, 4.09%) were the less common. The top three constitution in middle aged wereqi-deficiency (33 patients, 27.27%), phlegm-heat (23 patients, 19.01%) and blood stasis (22 patients, 18.18%). The top three constitution in the aged groupwere qi-deficiency (107 patients, 29.08%), yin-deficiency (76 patients, 20.65%) and blood-stasis (67 patients, 18.21%). Conclusions Stroke is closely related with imbalance of TCM constitution. The prevention and treatment of stroke patients with different ages should vary according to the constitutions.