Objective To investigate the changes of right ventricular(RV) volume and function in patients with atrial septal de-fect(ASD) .Methods 31 patients with ASD-type secundum(ASD group) and 30 age-matched controls(control group) were includ-ed .Single beat real-time three dimensional echocardiography (sbRT3DE) were performed in all to evaluate right ventricular volume and function ,then right venrtricular cardiac output(RVCO) were calculated with heart rate .Further correlative analysis was calcu-lated between right ventricular volume and function parameters of the ASD group and ASD size and the patient′s age .Results Right ventricular end-systolic volume(RVESV) ,right ventricular end-diastolic volume(RVEDV) ,right ventricular stroke volume (RVSV) ,right ventricular ejection fraction(RVEF) ,RVCO of the ASD group were significantly higher than that of control group (P<0 .001) .The maximum diameter of ASD had positive relation with RVESV ,RVEDV ,RVSV ,RVCO(r=0 .55 ,P<0 .01 ;r=0 .61 ,P<0 .01 ;r=0 .63 ,P<0 .01;r=0 .61 ,P<0 .01 ,respectively ) ,and was best correlation with RVSV .But ,there was no signifi-cant correlation between the maximum diameter of ASD and RVEF (r=0 .00 ,P>0 .05) .The patients′age had positive relation with RVESV ,RVEDV ,RVSV(r=0 .53 ,P<0 .01 ;r=0 .48 ,P<0 .01 ;r=0 .41 ,P<0 .01 ,respectively) ,had negtive relation with RVEF (r= -0 .44 ,P<0 .05) ,while there was no significant correlation between the patient′s age and RVCO(r=0 .26 ,P>0 .05) .Conclu-sion sbRT3DE is a fast and accurate new technology in evaluating RV volume and function .Atrial level left-to-right shunt leading to an increase in RV volume ,RV enlargement produces myocardial contractility .RV volume increases with ASD size ,then RV func-tion was enhanced .RV volume increases with patient′s age ,RV function is relatively lower .

Objective In order to evaluate the efficacy of esophageal varices ligation( EVL) , ultrasonic microprobe (UMP) was used to observe the esophageal varices and their collaterals before and after EVL. Methods Sixty patients merely with esophageal varices underwent ultrosonography via UMP before and 4,8,12 weeks after EVL and were divided into three groups:( Ⅰ )esophageal varices(EV) ( Ⅱ )esophageal and paraesophageal vein(PEV) varices( Ⅲ ) esophageal, para esophageal and perforating vein( PV) varices. Results In the following-up, the effective rate of type Ⅰ , Ⅱ , Ⅲ were 75% , 25% , 0% ; The recurrence rate 16% , 75% , and 100% respectively. PEV occurred in 12 out of 24 cases of group Ⅰ and broadened in all of the cases of group Ⅱ and Ⅲ; PV occurred in 11 out of 20 cases of group Ⅱ and all cases in groups M , and broadened in all cases of group Ⅲ. Conclusion Endoscopic ultrasonography can be served as guidance in selecting therapeutic measure in treating EV. EVL is merely suitable in treating EV; in EV patients with PEV and/or PV measures other than EVL should he selected.

Objective To evaluate the changes of right ventricular (RV) volume and function by using single beat real-time three dimensional (3-D) echocardiography in patients with atrial septal defect (ASD) before and after percutaneous closure. Methods During the period from July 2011 to Oct. 2013, a total of 45 patients with pure ostium secundum defect were admitted to authors’ hospital to receive percutaneous transcatheter closer. The patients were divided into ASD without pulmonary hypertension (PH) group (group A, n = 28) and ASD with PH group (group B, n = 17). By using 3-D echocardiography and right cardiac catheterization, the right ventricular end-diastolic volume (RVEDV), right ventricular end-systolic volume (RVESV), right ventricular stroke volume (RVSV), right ventricular ejection fraction (RVEF), right ventricular cardiac output (RVCO), pulmonary artery systolic pressure (PASP) and the mean pulmonary artery pressure (mPAP) were determined before and after the percutaneous transcatheter closer. The results were compared between the two groups. Results After the treatment a statistically significant reduction in RVEDV, RVESV, RVSV and RVCO were seen in all patients (P < 0.05). In group A, RVEF decreased significantly after ASD closure when compared with that determined before transcatheter closer (P < 0.05), while no significant reduction in RVEF was seen in group B (P > 0.05). Pulmonary artery pressure (PAP) decreased significantly in group B after ASD closure when compared with that obtained before Objective To evaluate the changes of right ventricular (RV) volume and function by using single beat real-time three dimensional (3-D) echocardiography in patients with atrial septal defect (ASD) before and after percutaneous closure. Methods During the period from July 2011 to Oct. 2013, a total of 45 patients with pure ostium secundum defect were admitted to authors’ hospital to receive percutaneous transcatheter closer. The patients were divided into ASD without pulmonary hypertension (PH) group (group A, n = 28) and ASD with PH group (group B, n = 17). By using 3-D echocardiography and right cardiac catheterization, the right ventricular end-diastolic volume (RVEDV), right ventricular end-systolic volume (RVESV), right ventricular stroke volume (RVSV), right ventricular ejection fraction (RVEF), right ventricular cardiac output (RVCO), pulmonary artery systolic pressure (PASP) and the mean pulmonary artery pressure (mPAP) were determined before and after the percutaneous transcatheter closer. The results were compared between the two groups. Results After the treatment a statistically significant reduction in RVEDV, RVESV, RVSV and RVCO were seen in all patients (P < 0.05). In group A, RVEF decreased significantly after ASD closure when compared with that determined before transcatheter closer (P < 0.05), while no significant reduction in RVEF was seen in group B (P > 0.05). Pulmonary artery pressure (PAP) decreased significantly in group B after ASD closure when compared with that obtained before transcatheter closer (P < 0.05). Conclusion Single beat real-time 3-D echocardiography is a newly-developed technique. This technique can quickly and accurately assess the right ventricular volume and function. Right ventricular volume will decrease after ASD closer. In ASD patients without PH the right ventricular function will decrease after ASD closer, while in ASD patients with PH the right ventricular function shows no changes after ASD closer although their PAP will decrease.

Objective To analyze the patterns of amino acid changes in liver failure patients treated with non-bioartificial liver support system (ALSS), and to explore the efficacy of ALSS in liver failure treatment. Methods A total of 146 liver failure patients treated with ALSS from June 2009 to August 2010 were recruited in this study. Paired blood samples were collected from every patient and serum amino acids and ammonia were tested by automatic amino acid analyzer. The changes of amino acids in patients with different prognoses, different types/phases of liver failure were evaluated.Measurement data were compared by paired t test. Results After ALSS treatment, liver failure patients experienced a significant decrease in serum glutamic acid and lysine [(395.62±200.24)μmol/Lvs (260. 05±169.56) μmol/L and (436. 73±326. 18)μmol/L vs (407. 12±292.01) μmol/L,respectively; t= 8. 611 and 2. 659, respectively; both P＜0.01)], while experienced greatly increases in threonine and branched-chain amino acids/aromatic amino acid ratio [( 1302. 90 ±1288.70) μmol/L vs (1406.70 ±1272. 34) μmol/L and 1. 23 ± 0. 53 vs 1. 36 ± 0.57, respectively; t = 2. 895 and 1. 061,respectively; both P＜0. 01)]. The changes of glutamic acid, tyrosine, arginine and methionine before and after ALSS treatment in patients with different prognoses, different types/phases of liver failure were all significantly different. Conclusions ALSS treatment could improve the serum amino acid disorder in liver failure patients. The amino acids in patients with different types/phases or different prognoses of liver failure change significantly after ALSS treatment.

Objective To establish a model for preparation of tissue-engineered skin grafts with hTERT cells carrying human papillomavirus type 6 (HPV 6) genome in vitro, so as to lay a foundation for studying HPV life cycle. Methods The full-length linear HPV6 genome and plasmid pEGFP-▲EGFP were electrophoretically cotransferred into hTERT cells. After selection using G418 resistance, Southern blotting was performed to determine the viral load of HPV6 in transfected cells. 3T3 J2 trophoblastic cells, type I rat-tail collagen and hTERT cells containing the full-length HPV6 genes (HPV6.hTERT cells)were mixed and cocultured on metal meshes to form skin graft-like structures. Hematoxylin and eosin (HE)staining was performed to observe the structure of formed skin grafts, an immunohistochemical assay to measure the expression of HPV6 L1 protein, and electron microscopy to observe virus particles in the skin grafts. Results The linear HPV6 gene was successfully transferred into hTERT cells, and Southern blotting showed the presence of HPV6 DNA in the transferred hTERT cells. The HPV6.hTERT cells, which were cocultured with 3T3 J2 trophoblastic cells and type I rat-tail collagen, proliferated and differentiated over time, and gradually formed skin grafts giving the appearance of verrucous hyperplasia. HE staining showed that the cocultured HPV6.hTERT cells could form typical stratified structure of skin after 7 days of cultivation, and histopathologic features of HPV infection, including obvious papillomatous hyperplasia, presence of vesicular cells, hyperkeratosis and parakeratosis, could be observed after 21 days. The immunohistochemical assay showed the expression of HPV6 L1 protein in the upper portion of skin grafts, and electron microscopy revealed the presence of HPV6 virus particles in skin grafts. Conclusions The established model for preparation of tissue-engineered skin grafts using HPV 6 genome-carrying cells provides a basis for biological studies of HPV, but its application is limited to some degree.

Case-oriented Workshop training mode is a professional training method which has been highly praised by anesthesiologists in recent years. It is of great significance to stimulate the participation of anesthesiologists and strengthen the application of the extended focused assessment with sonography for trauma (EFAST) in perioperative anesthesia management. The training of EFAST is divided into two stages: the first stage is about the relevant knowledge of EFAST and watching standardized operations on site, and the second stage is for each trainee to practice the operations under the guidance of the teacher until they can complete them independently. The EFAST adopts Workshop training mode, and the trainees can complete the target area scanning in about 90 seconds on average. It can maximize the training effectiveness and comprehensively improve the trainees' mastery of knowledge and skills in an all-round way. It is a training mode worthy of promotion in anesthesiology department.

OBJECTIVETo construct a universal, highly attenuated orf virus expression vector for exogenous genes using green fluorescent protein (GFP) as the reporter gene.

METHODSThe flanking regions of the ORFV132 of orf virus DNA were amplified by PCR to construct the shuttle plasmid pSPV-132LF-EGFP-132RF. The shuttle plasmid was transfected into OFTu cells and GFP was incorporated into orf virus IA82Delta 121 by homologous recombination. The recombinant IA82Delta121-V was selected by green fluorescent signal. The deletion gene was identified by PCR and sequencing. The effects of ORFV132 knockout were evaluated by virus titration and by observing the proliferation of the infected vascular endothelial cells in vitro.

RESULTSThe recombinant orf virus IA82Delta121-V was obtained successfully and quickly, and the deletion of ORFV132 did not affect the replication of the virus in vitro but reduced its virulence.

CONCLUSIONGreen fluorescent protein is a selectable marker for rapid, convenient and stable selection of the recombinant viruses. Highly attenuated recombinant orf virus IA82Delta121-V can serve as a new expression vector for exogenous genes.

Cells, Cultured ; Endothelial Cells ; metabolism ; Gene Deletion ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Orf virus ; Plasmids ; Transfection

Ultrasound-guided thoracic nerve block plays an important role in anesthesia and analgesia, but it is not easy to be mastered. This article discusses the application value of flipped classroom combined with 3D Body anatomy software for anesthesia specialty residents to learn ultrasound-guided thoracic paravertebral nerve block. This innovative education model includes three parts: before class, during class and after class. Before class, the teachers study the syllabus and make teaching micro-videos, and the cross-sectional anatomy added into the 3D Body anatomy software helps the residents understand. During class, teachers divide the residents into groups to report and exchange their learning experience, organize discussions, and finally give them comments. After class, the instructor will assign homework, assess the residents, and evaluate their satisfaction. This teaching model helps residents master the ultrasound-guided thoracic nerve block, and obtains satisfactory evaluation from the trainees, which is helpful for promotion.

Objective: To study the effect of low-to-moderate dose glucocorticoid therapy on viral clearance time in patients with COVID-19. Methods: A total of 72 patients diagnosed with COVID-19 from January 19 to February 17, 2020 at the First Affiliated Hospital, School of Medicine, Zhejiang University were recruited. All patients received oral abidol and/or combined lopinavir/ritonavir, darunavir antiviral, and symptomatic supportive care. Among them, 51 patients received methylprednisolone (0.75-1.50 mg·kg^-1·d^-1) (glucocorticoid treatment group), and 21 patients who did not use glucocorticoid were the control group. The time of stable virologic conversion insputumand the time of radiologic recovery in lungsince onset were compared between the two groups and among the normal patients.The Kruskal-Wallis test or Fisher exact test was used to compare the difference between groups. Results: The median ages of the glucocorticoid group and the control group were 52 [interquartile range (IQR):45, 62] years and 46 (IQR: 32, 56)years, and the differences were significant (P<0.05). The clinical conditions at hospital admission were different between the two groups (P<0.01). There were 52.0% critical ill patients in the glucocorticoid treatment group, compared to that of 71.4% normal patients in the control group. The median times from the onset tostable virologic conversion to negative in the two groups were 15 (IQR:13,20) days and 14 (IQR:12,20) days (P>0.05), and the difference was no statistically significant. The median times from onset to radiologic recovery were 13 (IQR: 11,15) days and 13 (IQR:12,17) days in the two groups, and there was no difference (P>0.05). In ordinary patients, the median timesfrom the onset tostable virologic conversion insputum were no difference (P>0.05), with 13 (IQR:11,18) days in the glucocorticoid group and 13 (IQR:12,15) days in the control group; The median times from onset to radiologic recovery in lungwere also no difference (P>0.05), with 12 (IQR: 10,15)days in the glucocorticoid group and 13 (IQR: 12,17) days inthe control group. Conclusions Low-to-moderate glucocorticoid treatment has no effect on the time of virus clearance in patients with different clinical types of COVID-19. The glucocorticoid is not recommended since no effectiveness on accelerating the improvement of radiologic recovery in lung has been observed.

Abstract OBJECTIVE To develop LC-MS method for the simultaneous determination of impurities A, C, and D of caspofungin acetate. METHODS Waters CORTECS® C18+(4.6 mm×150 mm, 2.7 μm) was used as the chromatography column. Mobile phase A and B were 0.1% formic acid-H2O and 0.1% formic acid-CH3CN, respectively. Electrospray ion source-single quadrupole mass spectrometry was used to detect impurities A and C in positive ion mode and impurity D in negative ion mode. RESULTS The correlation coefficient r was ≥ 0.999 in linearity ranges of impurities A, C and D. The average recoveries were 100.5%, 104.1% and 105.2%, respectively, with RSD<4%(n=6). The LOQs (S/N=10) of impurities A, C and D were 31.8, 6.99 and 15.5 ng·mL-1 respectively. The contents of impurities A, C and D in the three samples were all below the limits. CONCLUSION The developed LC-MS method is simple, sensitive, and applicable, which can be used to simultaneously determine impurities A, C and D in caspofungin acetate and can also provide a reference for the detection of other impurities in caspofungin acetate.