Abstract: Objective To establish a method for detecting sdaB virulence gene of Streptococcus pyogenes with loop mediated isothermal amplification (LAMP). Methods According to the conserved sequence of Streptococcus pyogenes sdaB gene published in GenBank (GenBank: 69901515), LAMP primers were designed with Primer Explorer V5.0 software. Main components of LAMP reaction system were optimized including of fluorescent dye, MgSO4, betaine, deoxyribonucleosidetriphosphate (dNTP), and Bst DNA polymerase, with the concentration of MgSO4 from 0 mmol/L to 12 mmol/L, betaine from 0 mol/L to 2.4 mol/L, dNTP from 0.2 µmol/L to 2 µmol/L, forward inner primer (FIP) and backward inner primer (BIP) from 0.2 µmol/L to 2 µmol/L respetively, forward outer primer (F3) and backward outer primer (B3) from 0.2 µmol/L to 0.4 µmol/L, Bst DNA polymerase from 0.16 U/µL to 0.96 U/µL, fluorescent dye from 0.2 µmol/L to 2 µmol/L. With the optimized system, the methodological specificity and the minimum detection limit were evaluated on the ABI7500 real-time fluorescent quantitative PCR analyzer, and 13 standard strains including Group A Streptococcus, Group B Streptococcus, Group C Streptococcus, Group G Streptococcus, Streptococcus pneumoniae, Streptococcus viridis, Enterococcus faecalis, Enterococcus faecium, Neisseria gonorrhoeae, Lactobacillus acidophilus, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa were detected. Finally, 103 clinical samples were tested. Results The optimized reaction system contained 25 µL reaction mixture, including 0.8 µL of 25 µmol/L fluorescent dye, 1 µL of 100 mmol/L MgSO4, 6 µL of 5 mol/L betaine, 1.4 µL of 25 mmol/L dNTP, 2 µL of 20 µmol/L FIP and BIP, 0.5 µL of 20 µmol/L F3 and B3, 1 µL of 8 U/µL Bst DNA polymerase, and 2 µL of template. After adding deionized water, the mixture was incubated at 63°C for 45 min to complete the reaction. The limit of detection (LOD) was 500 pg/µL. All 12 non-S. pyogenes strains tested were negative. Compared with the culture method, the clinical sensitivity and specificity were 100.0% (16/16) and 96.6% (84/87), respectively, for 103 clinical samples. Conclusions This LAMP assay is reliable for the detection of Streptococcus pyogenes in clinic and is suitable for field detection with good specificity and sensitivity, as well as simply operation.

Objective:To optimize 3.0T MRI diagnosis indicator in breast cancer and to select the best MRI scan program.
Methods:Totally 45 patients with breast cancers were collected, and another 35 patients with benign breast tumor served as the control group. All patients underwent 3.0T MRI, including T1-weighted imaging (T1WI), fat suppression of the T2-weighted imaging (T2WI), diffusion weighted imaging (DWI), 1H magnetic resonance spectroscopy (1H-MRS) and dynamic contrast enhanced (DCE) sequence. With operation pathology results as the gold standard in the diagnosis of breast diseases, the pathological results of benign and malignant served as dependent variables, and the diagnostic indicators of MRI were taken as independent variables. We put all the indicators of MRI examination under Logistic regression analysis, established the Logistic model, and optimized the diagnosis indicators of MRI examination to further improve MRI scan of breast cancer.
Results:By Logistic regression analysis, some indicators were selected in the equation, including the edge feature of the tumor, the time-signal intensity curve (TIC) type and the apparent diffusion coeffcient (ADC) value when b=500 s/mm2. hTe regression equation was Logit (P)=-21.936+20.478X6+3.267X7+21.488X3.
Conclusion:Valuable indicators in the diagnosis of breast cancer are the edge feature of the tumor, the TIC type and the ADC value when b=500 s/mm2. Combining conventional MRI scan, DWI and dynamic enhanced MRI is a better examination program, while MRS is the complementary program when diagnosis is diffcult.

In order to express the gene of LEN-5 β-lactamase from a Klebsiella pneumoniae strain,plasmids in the strain were extracted and an 879bp product of LEN-5 gene was obtained with PCR.After being digested with Nde I and Xho I,LEN-5 gene was cloned into pET-26b (+) vector.Then it was confirmed by digestion and DNA sequencing in recombinant plasmid before transformed into E.coli BL21 (DE3).After inducing by IPTG,LEN-5 β-lactamase was expressed.Protein extraction was processed by ultrasonic and protein activity was detected by nitrocefin.The isoelectric focusing electrophoresis showed a pI of 7.6.These results indicated that the LEN-5 gene has been cloned and expressed in prokaryote cell successfully.

Objective:To investigate the effects of acupuncture therapy on acute cerebral infarction. Methods: Eighty patients with acute cerebral infarction were randomly divided into a treatment group of 40 cases and a control group of 40 cases. Xuanzhong(GB 39)-through-Sanyinjiao(SP 6) acupuncture was performed as a main treatment. The curative effects were compared between the two groups and the sizes of cerebral infarct, between pretreatment and posttreatment after one course of treatment. Results: The total recovery rate was 88.5% in the treatment group and 57.5% in the control group after one course of treatment. There was a significant difference between the two(P<0.05). The rate of change in the infarct for the better was significantly higher in the treatment group than in the control group. There was also a significant difference(P<0.05). Conclusion: This treatment is an effective method for lowering the rate of apoplectic disability and raising the cure rate.

Objective　To investigate the relationship betwee n tricuspid insufficiency (TI) and the function of the right ventricle, degree o f pulmonary vascular pathological change in patients with rheumatic heart diseas e so as to provide the indication of operation for treating TI. Methods 　41 cases of rheumatic heart disease with TI accompanying with bicuspid pathological changes were reviewed. Correlation analysis was done between the s everity of TI and the right ventricular size(RVS), pulmonary artery pressure(PAP ), pulmonary artery resistance(PAR) etc. Results　It was found t hat the size of the right ventricle, PAP, PAR were positively correlated to th e degree of TI. Tricuspid annuloplasty should be carried out in patients with RV D>40 mm, PAR >48 kPa*s/L, PAP>8 kPa. Conclusion　TI resulted ma inly from insufficient function of the right ventricle and marked patholog ical changes of the pulmonary blood vessels. Doppler echocardiography evaluation for the function of right ventricle and pathological condition of pulmonary blo od vessels might be of significant in deciding whether tricuspid annuloplasty sh ould be performed simutaneously in patients of bicuspid valve replacement.

Objective To summarize the individualized cavity Single branch stent grafting through rebuilding the aortic arch surgery in 26 cases of the application of the Stanford type A aortic dissection. Methods From 2010 January to 2014 October, 26 patients received Stanford type A aortic dissection surgery, 26 patients received individualized cavity single branch stent grafting to rebuild the aortic arch surgery , together with improved myocardial protection fluid. Results In the present study, 26 cases with aortic dissection that were treated with single branch stent grafting for the reconstruction of aortic arch under DHCA and selective cerebral perfusion. Twenty-six patients received individualized cavity single branch stent grafting reconstruction of aortic arch surgery alone, and were stopped by using deep cryogenic loop (DHCA) plus selective cerebral perfusion surgical treatment. One patient suffered from permanent nerve dysfunction iand give up treatment. Conclusion The sexua branch stent grafting in reconstruction of aortic arch operation could simplify the operation procedures , shorten the operation time, and reduce the amount of blood transfusion and postoperative drainage.

Objective To observe the recent clinical effect of application of individual open fenestrated stent graft in type A aortic dissection without developing the greater curve of the arch. Method From December 2014 to November 2016,15 patients of type A aortic dissection without developing the greater curve of the arch un-derwent endovascular total arch replacement using individual open fenestrated stent graft in the Anhui Province Hospital.Among them,8 cases were only operated with open fenestrated stent graft in aortic arch, 7 cases with open fenestrated stent graft in aortic arch added 1 or 2 small stent graft.Result There was 1 postoperative death caused by severe low cardiac output. The rest of the patients were successfully discharged from the hospital, without ner-vous system and related complications. Follow-up computerized tomographic angiography showed all implanted stents were wide expansion and in a good position. No endoleaks and thrombus obliterated of the corresponding false lumen was found. Conclusion Individual open fenestrated stent graft is suitable for type A aortic dissection without developed the greater curve of the arch.Its significantly simplify the total arch replacement operation steps, reduce anastomotic and shorten the lower body arrest time. Consequently, reduce the risk of operation difficulty, postoperative blood loss and other viscera damage probability significantly. The early and middle term clinical re-sults is satisfactory.

Objective To explore the impact of heat shock factor 2 (HSF2) on the development of lung cancer by promoting interleukin (IL)-10 expression.Methods 50 lung cancer tissues and adjacent normal tissues selected from 50 patients,the expression level of mRNA and protein of HSF2 and IL-10 were respectively detected by RT-PCR,Western blot and Immunohistochemistry;To interfere with expression of HSF2 in A549 cells by siRNA,the expression level of IL-10 was detected by Western blot.Results Compared with the adjacent normal tissues,HSF2 of 76％ (38 of 50) cases were up-regulated (P＜0.01),IL-10 of 80％ (40 of 50) eases were up-regulated (P＜0.01),protein level consistent with mRNA level.The up-regulation expression of IL-10 in lung cancer tissues and HSF2 positively correlated (R2 =0.921 6).The expression of IL-10 in A549 cells was weakened through interference with HSF2 by siRNA.Conclusion HSF2 may contribute to the development of lung cancer by facilitating the expression of IL-10.

Objective To investigate the short-term efficacy and safety through analyzing the results of 11 cases of video assisted minimally invasive cardiac surgery. Methods Eleven patients who had underwent video assisted minimally invasive cardiac surgery in the second hospital of Anhui medical university from November 2017 to August 2018 were retrospectively analyzed. One patient underwent simple atrial septal defect repair, 6 patients underwent atrial septal defect repair+tricuspid valve repair, 3 patients underwent mitral valve replacement, and 1 patient underwent mitral valve repair + tricuspid valve repair. Results All patients underwent video cardiac surgery without death, with no major bleeding (need secondary thoracotomy to stop bleeding) and no neurological complications. One patient of incision fat liquefaction healed after dressing change. One patient developed lower limb artery stenosis, which was treated conservatively and discharged after hospitalization. Conclusions Video assisted minimally invasive cardiac surgery can be safely applied to simple congenital heart disease and valve surgery as long as patients are strictly screened. Due to the advantages of small trauma, less bleeding, and quick recovery, it is worth promoting.

OBJECTIVETo compare the clinical efficacy between total aortic arch reconstruction with a individualized combined branched stent grafting technique and total aortic arch replacement combined with stented elephant trunk implantation for patients with Stanford A aortic dissection.

METHODSTotally 44 patients with Stanford A aortic dissection treated with surgical treatment from January 2007 to July 2013 were included in this study. The patients were divided into two groups. Group A (n = 22) patients were treated by total arch replacement with stented elephant trunk procedure. Group B (n = 22) patients received individualized combined branched stent grafting technique. Age, gender and disease severity were similar between the two groups (all P > 0.05). Echocardiography and aortic CT angiography were performed pre-operation and at 1 month after operation.

RESULTSOperation was successful in all 44 patients. Cardiopulmonary bypass time, aortic cross clamp time, circulation arrest time and duration of ventilator assisted breathing were significantly longer, postoperative drainage volume and blood transfusion volume were significantly larger and hospitalization cost was significantly higher in group A patients compared those in group B patients (t = 2.791 to 43.465, all P < 0.05). One month after operation, the maximum internal diameter of aorta was smaller than pre-operation in both group A ((33 ± 1) mm vs. (45 ± 6) mm, t = 10.076, P = 0.000) and group B ((33 ± 2) mm vs. (45 ± 8) mm, t = 5.979, P = 0.000) . Left ventricular ejection fraction had no significant difference before and 1 month after operation in both groups (P > 0.05).

CONCLUSIONThe total aortic arch reconstruction with individualized combined branched stent grafting technique is technically easier, shortens the operation time, reduces the blood transfusion volume compared to the classical aortic arch operation.

Adult ; Aneurysm, Dissecting ; surgery ; Aortic Aneurysm, Thoracic ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Stents ; Treatment Outcome