Objective To clone the genes coding for cysteine proteases (CPs, TvCPs) from Trichomonas vaginalis and to analyze their genetic variations with the related sequences from NCBI database (GenBank) and T. vaginalis Genome Project database from The Institute for Genomic Research (TIGR). Method TvCP genes were amplified using PCR, and inserted into vector pET28b or pBS-T. The recombinant plasmids were then transformed to Escherichia coli BL21 or Top10 strain. The recombinant plasmids were used for sequencing. Homologous TvCP genes were blasted based on NCBI GenBank and TIGR T. vaginalis Genome Project database. The sequences of cloned TvCP genes were aligned and clustered by Clustal X (1.83 version) with retrieved sequences. Comparisons of amino acids among cathepsin L-like TvCPs, human L-like cathepsins and papaya papain were performed using DNAstar software, and their phylogenic tree was constructed based on neighbor-joining method using Clustal X. Results Two TvCP3 clones and one TvCP2 had a high identity of more than 99% with their responding TvCPs. Three clones of TvCP4 genes, GZ-CP4-clone 1-3, belonged to two members of a family showing a high percentage identity of more than 97.5% with the sequences of TvCP4 genes from databases (GenBank and TIGR) both at amino acid and nucleotide levels. Nine homologous TvCP4 pro-enzymes with 304 amino acids and other two members with deletions of N-terminal sequence existed in T. vaginalis sharing a similarity of 62.3-96.7% amino acids, which may evolve by means of gene replication and deletion. TvCP1-4, TvCP12, TvCP25 and CP65 had an identity of 61-88.2% at amino acid levels. So far, all reported sequences of C1 family from T. vaginalis belonged to capanthesin L-like subfamily with the same enzymatic active sites, conserved cysteine residues and similar structural features such as ERFNIN-like motif in pro-enzyme region, suggesting that they might result from gene duplication and mutations. Conclusion TvCPs belong to cathepsin L-like family with genetic diversity, but they have the same active amino acid residues, cysteine residues and similar structural characteristics, suggesting that they may derive from one ancestor.

Diagnosis, Differential ; Ependymoma ; metabolism ; pathology ; Female ; Hemangioblastoma ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Meningeal Neoplasms ; metabolism ; pathology ; surgery ; Meningioma ; metabolism ; pathology ; surgery ; Middle Aged ; Mucin-1 ; metabolism ; Neoplasm Recurrence, Local ; Vimentin ; metabolism

The genetic diversity of three Tibetan herbs, i. e., Sang-Di, E-Dewa and Ye-Xingba (Tibetan names), was studied based on the field collection, specimen identification and DNA sequence analysis. Swertia hispidicalyx, Gentiana lhassica and Scrophularia dentata, as the original plants of the three Tibetan herbs, were collected and identified. The regions of ITS, matK, rbcL, rpoC1, trnL(UAA), psbA-trnH, atpB-rbcL, trnS (GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF(GAA) and nadl 2nd intron were amplified and sequenced. The ITS regions of S. hispidicalyx and S. dentata were cloned and sequenced, and the sequences were classified into different genotypes. All the sequences were analyzed and compared with those of closely related species. Our studies may provide reference for the genetic diversity analysis and molecular identification of the three Tibetan herbs.
Genetic Variation ; Gentiana ; classification ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Plants, Medicinal ; classification ; genetics ; Scrophularia ; classification ; genetics ; Swertia ; classification ; genetics ; Tibet

Objective Few studies are reported on the radicular groove and root canal variation of the mandibular first premolar.This study aimed to observe the root canal system and root variation of the mandibular first premolar by cone-beam computed tomography (CBCT).Methods We selected CBCT images of 643 mandibular first premolars of 336 patients and subjected the data obtained to statistical analysis.Results Of the 643 mandibular first premolars, 637 (99.1％) had a single root and the other 6 had double roots;502 (78.1％) had a single root canal, 133 (20.7％) had two, 5 (0.78％) had three, and 3 (0.47％) presented C-shaped root canal morphology.The average length of the 643 mandibular first premolars was (13.2±1.5) mm and the mean distance from the apical foramen to the anatomical apical end of the root was (0.93±0.35) mm.Totally, 123 (19.1％) of the mandibular first premolars had a radicular groove, of which 94 (76.4％) had double root canals and the root canal systems included typesⅠ in 24 (19.5％), Ⅱ in 2 (1.6％), Ⅲ in 19 (15.4％), Ⅳ in 1 (0.8％), Ⅴ in 69 (56.1％), and Ⅶ in 3 (2.4％).Conclusion The root canal system of the mandibular first premolar has a significant variation.CBCT can reveal the complex anatomical structure of the mandibular first premolar and provide reliable evidence for the root canal treatment.

Objective To investigate the clinical value of ~(99)Tc~m-ethylenedicysteine (EC) diuretic renography (DR) in pre-and post-operative pediatric congenital hydronephrosis.Methods The DR with injection of Furosemide at 15 min of forty children with hydronephrosis was retrospectively studied.The preoperative renal blood perfusion rate (BPR),effective renal plasma flow (ERPF),grade of hydronephrosis,renogram and renal dynamic imaging of pre- and post-operative kidneys were compared.The t-test and Mann-Whitney test were used for data analysis.Results (1) Of 40 pathological kidneys,the BPR increased 5.99% (t=-5.13,P＜0.01)from pre-operative to post-operative:(34.05±11.07)% to (40.04±8.56)%.The ERPF increased 12.48 ml/min(t=-4.35,P＜0.01) from pre-operative to post-operative:(57.81±34.32)ml/min to(70.29±5.37)ml/min.(2)The grade of hydronephrosis of 40 pathological kidneys improved significantly(Z=-2.64,P＜0.01) with the mean sum of ranks of 47.21 pre-operatively to 33.79 post-operatively.(3) As the hydronephrosis worsened,the collecting system became bigger,the renal parenchyma became thinner,the extent of intrarenal parenchymal photopenia became larger and the response to diuretic challenge in pathological kidneys decreased or became totally irresponsive.(4)Thirty-seven cases of obstruction at ureteropelvic junction (UPJO) and 3 cases at ureterovesical junction (UVJO) were diagnosed by DR,which were all confirmed by surgery.Conclusions DR is a reliable method to evaluate pediatric congenital hydronephrosis.It can accurately reflect the grade and (or) severity of the disease,guide therapy and assess the therapeutic success of operation.

Objective: To analyze clinical characteristics and risk factors in patients with acute myocardial infarction (AMI) complicated with cardiac rupture (CR) and to explore the prevention and treatment strategy in clinical practice. Methods: A case control study was conducted in 2 groups: CR group, the patients with coronary angiography conifrmed AMI with CR,n=44 and Control group, the patients with simultaneous STEAMI and by 1:3 pair-matched ratio,n=132. Clinical information was compared between 2 groups and the relevant risk factors for predicting CR were studied by Logistic regression analysis. Results: Compared with Control group, CR group had the lower ratio of β-receptor blocker application (22.7% vs 81.4%),P<0.05. Univariate regression analysis indicated that lower body mass index, incipient MI, anterior MI, no-reperfusion therapy, delayed reperfusion therapy, lower blood pressure at admission, post-infarction angina, ventricular aneurysm, higher Gensini score, high blood levels of cretinine and BNP, low ejection fraction were the risk factors for CR occurrence in STEAMI patients, allP<0.05. Multivariate regression analysis presented that incipient MI (P<0.049, OR=7.462), post-infarction angina (P<0.000, OR=8.591), ventricular aneurysm (P<0.005, OR=4.617) and higher Gensini score (P<0.001, OR=2.788) were risk factors for CR occurrence in STEAMI patients. Conclusion: Incipient MI, post-infarction angina, ventricular aneurysm and higher Gensini score are the risk factors for CR occurrence in STEAMI patients.

Objective To evaluate the short-term clinical efficacy and safety of administration of levosimendan or milrinone added to conventional therapy in patients with decompensated heart failure.Methods A total of 180 patients admitted due to heart failure [NYHA (New York Heart Association) class Ⅲ or Ⅳ] were randomly (random number) divided into control group,milrinone group and levosimendan group (n =60,each group).A continuous infusion of milrinone added to conventional therapy was administered for 72 hours in milrinone group,while administration of levosimendan for 24 hours in levosimendan group.The changes in left ventricular ejection fraction (LVEF),left ventricle end-diastolic diameter (LVDD) and B-type natriuretic peptide (BNP) plasma level were compared between before and after treatment,respectively,and comparisons of improvement in cardiac function (NYHA class) and hospital mortality were carried out among three groups.Patients were further followed up at 3 months after treatment.Results The LVEF in levosimendan group after treatment had significantly more increased than that in control group [(32.0±6.3)％ vs.(30.6 ±5.5)％,P =0.007].Compared BNP before treatment,the sums of BNP deducted were 444.0 (-74.0,1068.0) pg/mL,469.0 (141.5,1151.5) pg/mL and 936.5 (437.8,1566.8) pg/mL in control group,milrinone group and levosimendan group,respectively after treatment (all P ＜ 0.01).Moreover,the deduction in BNP was more dramatic in levosimendan group compared with control or milrinone group (t =3.256 or 2.665,P =0.004 or 0.026).After treatment for 5 days,the probability at least of achieving more effectively better improvement in NYHA class (cardiac function) in levosimendan group was 2.036 times that of control group (95％ CI:1.030-4.028,P =0.041).The incidence of combined end point events (death or readmission) in levosimendan group was significantly lower than that in milrinone group (50％ vs.70％,HR =0.573,95％ CI:0.358-0.917,P=0.020),while in hospital mortality,readmission or 3-month mortality incidence was similar among 3 groups (P ＞ 0.05).Conclusions The short-term clinical efficacy of levosimendan is superior to that of milrinone or conventional therapy in patients with decompensated heart failure.

Objective To investigate the relationship between regular exercise habit and coronary collaterals of patients with acute coronary syndrome (ACS).Methods TWo hundred and thirty-night patients diagnosed ACS and operated coronary angiography (CAG) showing severe coronary stenosis were enrolled hospitalized from May 2012 to October 2012.They were divided into regular exercise group (n =102) and irregular exercise group(n =137) according to the exercise frequency.The information of the general data,the information of CAG and other relevant index were collected.The coronary artery score was recorded according to the Censini and the coronary collateral class was made according to the Rentrop.Other characters in clinical and laboratory were recorded.Multi-factor regression analysis was used to analysis the influence factors of coronary collateral.Results The proportion of coronary collaterals (41.2％ (42/102)) in the regular exercise group was higher than that in the irregular exercise group (24.1％ (33/137)),and the difference was statistically significant(x2 =7.929,P =0.005).Lg(Gensini score) was (1.89 ± 0.18) the and (1.94 ± 0.19) in the regular exercise group,The left ventricular ejection fraction was 57.0％ (52.0％,60.0％) in the regular exercise group and 50.0％ (45.0％,57.0％) in the irregular exercise group,and the difference was statistically significant (Z =-5.152,P =0.000).Multi-factor regression analysis showed that regular exercise (OR =3.423,95％ CI:1.790-4.578),diabetes mellitus (OR =0.451,95％ CI:0.212-0.962),B-type natriuretic peptide (OR =2.412,95 ％ CI:1.271-4.578),non-ST-segment elevation ACS (OR =2.383,95％ CI:1.185-4.791),chest pain history (OR =2.207,95％ CI:1.175-4.145),Gensini score (OR =1.538,95％ CI:1.141-2.073) were independent influence factors of coronary collateral(P ＜ 0.05).After adjusting other factors,the patients with regular exercise had better coronary collaterals than that with irregular exercise (OR=3.423,95％CI:1.790-6.548,P ＜0.001).Conclusion The regular exercise can promote coronary collateral emergence for the patients with ACS.

Objective To evaluate the improvement effect of levosimendan by vein injection on short term cardiac function of patients with decompensated heart failure.Methods One hundred and sixty patients admitted due to heart failure were randomly divided into levosimendan group and control group (80 subjects for each group).Patients in control group were given a regular therapy including diuretics,vasodilators (including the recombinant human brain natriuretic peptide),angiotensin converting enzyme inhibitor(ACEI) or angiotensin Ⅱ receptorantagonists(ARB),β blockers,spironolactone and stain.Patients in levosimendan were administered levosimendan for 24 hours plus regular therapy.The improvements of dyspnoea in 9 days and cardiac function classification in 30 days after therapy were assessed.Mortality of 1 month and 3 month in two group were calculated and compared during follow-up.Results The dyspnoea improvement rate was superior than that of control group during 9 days (OR =1.956,95％ CI:1.156-3.310,P =0.013).The improvements in the levosimendan group were better than in the control group at 1 st day (OR =2.261,95 ％ CI:1.280-3.999,P =0.005),at 3rd (OR =2.002,95 ％ CI:1.111-3.607,P =0.021) and 5th day (OR =1.846,95 ％ CI:1.009 -3.377,P =0.047).However,there was no significant difference in term of improving dyspnoea between the levosimendan group and the control group at 9th day (P =0.126).Similarly,the improvement of cardiac function classification in the levosimendan group was superior than the control group during 30 days (OR =1.933,95％ CI:1.229-3.040,P =0.004).Although no significant difference was seen regarding of improving cardiac function classification between the two groups at 30th day after treatment (P =0.115),the improvements in the levosimendan group were better than in the control group at 3rd (OR =1.986,95％ CI:1.195-3.300,P =0.008),5th (OR =2.268,95 ％ CI:1.329-3.873,P =0.003),9th (OR =2.627,95 ％ CI:1.419-4.860,P =0.002) and 14th day(OR =2.212,95％ CI:1.189-4.112,P =0.012).Moreover,there was a nonsignificant reduction in terms of mortality in levosimendan group during 1-month and 3-month follow-up compared with control group (P ＞ 0.05).Condusion Levosimendan can effectively improve the short-term cardiac function in patients with decompensated heart failure.

Objective To evaluate the efficacy of intravenous recombinant human brain natriuretic peptide in acute anterior myocardial infarction complicated with heart failure.Methods Two hundred patients suffered from acute anterior myocardial infarction complicated with heart failure were randomly divided into two groups:rhBNP group ( n =100) and control group ( n =100 ).All patients were given conventional treatment,patients in rhBNP group were given rhBNP on the basis of conventional therapy.The clinical effectiveness including the improvement of cardiac function,cardiac ultrasound data,the incidence of hospital adverse cardiac events,and six month follow-up were compared between the two groups.Results The degree of decompensation and Killip class in rhBNP group were better than those of control group after treatment ( improved dyspnea:significantly improved:36 vs 27 ; improved:49 vs 46; no improvement:11 vs 20 ; deterioration:4 vs 7 ; Ridit value:0.4618 vs 0.5382,P =0.043) ( Killip class:significantly improved:26 vs 20; improved:56 vs 45; no improvement:14 vs 25 ; deterioration:4 vs 10; Ridit value:0.4553 vs 0.5447,P =0.017 ).After treatment for one week,The LVEF improvement in rhBNP group was more remarkable than that of control group ( [ 53.0 ± 5.2 ] ％vs.[ 50.0 ±:6.2 ] ％,P =0.014).The occurrence rate of angina ( 13.0％ vs.27.0％,P =0.013 ),heart failure ( 18.0％ vs.32.0％,P =0.022) and major adverse cardiac events(MACE) ( 17.0％ vs.30.0％,P =0.030) inrhBNP group was lower than that in control group.During 6 months follow-up period,event-free survival in rhBNP group was higher than that in control group ( 69.0％ vs.55.0％,P =0.041 ).Conclusion Transvenous injection of rhBNP combined with other routine treatment can improve cardiac function in patients with myocardial infarction in acute anterior myocardial infarction.It can also decrease adverse cardiac events during hospitalization and increase event-free survival in 6 months follow-up period.