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Objective:To explore the effect of systolic anterior motion (SAM) of mitral valves on the morphology and function of left ventricular outflow tract (LVOT) in patients with hypertrophic obstructive cardiomyopathy (HOCM) using computer fluid dynamics based on three-dimensional echocardiography with inverted grey values.Methods:A total of 40 patients with hypertrophic cardiomyopathy were divided into SAM group (24 cases) and non SAM group (16 cases) in Renmin Hospital of Wuhan University from April 2016 to October 2019. Two dimensional and three-dimensional echocardiographic data of the patients were collected. The LVOT morphological model was constructed based on the post-processing of three-dimensional echocardiography data, and the LVOT flow field model was constructed based on the time-volume curve of left ventricle. LVOT peak velocity was obtained to assess the agreement with echocardiography measurements. Area of LVOT, average velocity, flow rate and iso-surface area of vortex of different levels were obtained and compared between the two groups.Results:There was a good correlation between cardiac fluid model and echocardiographic measurement ( r=0.943, P<0.01). The Bland-Altman consistency interval was -75.0-111.3, and 92.5% of the points were within the consistency limit. Compared with non-SAM group patients, the peak velocity of LVOT increased, the area of LVOT decreased, the flow rate decreased and the area of vortex increased in SAM patients (all P<0.01). In the SAM group, in 16 patients the double orifice LVOT was observed due to the contact between mitral valve and septum, in 1 patient the single orifice LVOT structure was observed with contact between mitral value and septum, and in 7 patients, single orifice LVOT without contact between mitral value and septum. In SAM patients, compared with single orifice LVOT, patients with double orifice LVOT were observed with higher LVOT velocity, smaller LVOT area and higher vortex area with high level(all P<0.05). Conclusions:Accurate fluid models can be obtained using three-dimensional echocardiography with inverted grey values. In SAM patients, contact between mitral valve and septum leads to the formation of double orifice structure and the increase of vortex level in LVOT.
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Objective:To meet the individualized needs of residents in standardized residency training of cardiac intervention specialty for cardiac ultrasound teaching, to construct a bionic 3D printing model of ultrasound source, and to discuss its application value in ultrasound teaching and simulation exercises.Methods:Sixty residents majored in cardiovascular medicine receiving standardized residency training of ultrasound in ultrasound imaging department of Renmin Hospital of Wuhan University were randomized into experimental group and control group in average. The experimental group adopted ultrasound bionic 3D printing heart model combined with problem-based learning (PBL) teaching method, and the control group adopted traditional multimedia PPT combined with PBL teaching method. At the end of teaching activities, theoretical examination, operation assessment and questionnaire survey were conducted to evaluate and compare the teaching effect between the two groups. SPSS 21.0 software was performed for t test. Results:The theoretical examination scores of the experimental group and the control group were (81.4±8.2) points and (74.8±9.4) points, respectively, P=0.002, and the operation assessment scores were (89.1±5.6) points and (71.5±8.8) points, respectively, P<0.001. The questionnaire survey showed that the experimental group had better feedbacks than the control group in the aspects of mastery of heart anatomy knowledge, learning experience, learning interest, learning efficiency, understanding of interventional heart disease, clinical skills of interventional operation and overall satisfaction (all P<0.05). Conclusions:Ultrasound bionic 3D printing combined with PBL teaching method can help strengthen the mastery of cardiac anatomy knowledge, promote the learning experience of cardiac ultrasound and improve the skills of interventional operation. The teaching effect is significant and the students' satisfaction is high.
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Objective:To evaluate the accuracy and application value of the left atrial appendage(LAA) occlusion simulation in dynamic fluid in vitro based on left atrial appendage models. Methods:The preoperative two/three-dimensional transesophageal echocardiography(2D/3D-TEE) images of 21 patients with atrial fibrillation undergoing percutaneous LAA occlusion were retrospectively analyzed. LAA models with different materials (silicone-hydrogel LAA models, silicone LAA models and Tangoplus LAA models) were obtained by 3D printing. After simulation evaluation set models in a dynamic fluid system to simulate the occlusion operation, the type of occluder was predicted, the correlation between them and the final occluder used in the operation was analyzed.According to 2D/3D-TEE monitoring in 21 patients, the compression ratio of occluders was measured and the residual leakage around occluders was observed. With the same type occluder as in surgery, the compression ratio of occluders and residual leakage around the occluders in the three LAA models were observed and compared with intraoperative measurements, the descriptive and correlation analyses were conducted.Results:The simulation score of silicone-hydrogel LAA models was higher than those of silicone LAA models and Tangoplus LAA models. The occluder size in silicone-hydrogel LAA models was more correlated to the size used in the operation( r=0.937, P<0.001) than silicone LAA models ( r=0.918, P<0.001) and Tangoplus LAA models ( r=0.895, P<0.001). With the same size of occluder, there was no significant difference between the compression ratio of the occluder in silicone-hydrogel LAA models and intraoperative ( P=0.908). In 21 patients, 7 cases of residual leakage of occluder were observed during operation, while 4 cases were observed in silicone-hydrogel LAA, 5 cases were observed in silicone LAA models and 5 cases in Tangoplus LAA models. Conclusions:Preoperative simulation system of left atrial appendage occlusion based on silica gel-hydrogel model is helpful for preoperative decision-making, simulation exercise and post-operative evaluation of percutaneous left atrial appendage occlusion.
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Objective To evaluate the accuracy of simulated operation before the left atrial appendage ( LAA ) occlusion by filed LAA model with soft materials ,which is based on transesophageal three-dimensional echocardiography ( 3D-TEE) . Methods Silica and hydrogel were mixed to imitate cardiac tissue . Silica and hydrogel mixed materials ,silica and Tangoplus were subjected to mechanical detection and ultrasonic shear wave elastography ( SWE) . The preoperative and postoperative 3D-TEE image processing was performed on 21 patients undergoing LAA occlusion . The silica and hydrogel LAA 3D model ,silicone model and Tangoplus model were acquired to evaluate the accuracy of 3D model . LAA closure was simulated using the 3D models and the compression ratio of the device in models was compared with the value measured in operation . Results The silica and hydrogel mixed materials were successfully prepared . The elastic modulus and stress-strain curve were closer to the myocardial tissue . Twenty-one 3D printing models of three materials were obtained in 21 patients based on 3D-TEE images .There was no significant difference in the anchor zone between the 3D model and 3D-TEE ( P > 0 .05) .The 3D model measurements were consistent with the 3D-TEE measurements .The compression ratios of the decive in the 3D models of all three materials were greater than those measured in operation ,and the compression ratio of the device in the 3D model of silica and hydrogel materials had a better correlation with that measured in operation ( r =0 .900 ,P < 0 .01) .Conclusions The 3D model of LAA made of silica and hydrogel material based on 3D-TEE can improve the accuracy of the LAA occlusion simulation ,also can optimize the preparation of LAA occlusion .
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Objective To evaluate the feasibility and accuracy of three-dimensional (3D) printed left atrial appendage (LAA) models based on 3D transesophageal echocardiography (3D-TEE) data and the application value for treating special anatomic LAA.Methods Data of 18 patients of atrial fibrillation who were underwent LAA occlusion were retrospectively analyzed,including 3D-TEE and CT volume data of the patients.The 3D-TEE data of the LAA were post-processed and a flexible material was used to print the LAA model by 3D printer.The morphological classification and lobulated classifications of LAAs were assessed by the 3D printed models.The measurements of long axis,short axis and depth of LAAs were also performed.And the measurement and classification results were compared with those based on 3D-TEE and CT volume images.A occluder release test was performed on the 3D printed models for patients with challenging LAA morphology.Results For all 18 patients,3D-TEE full volume data of the LAA were successfully reprocessed and printed as 3D LAA models.The consistency of morphological classifications and lobulated classifications of LAAs based on 3D printed models and cardiac CT were 0.92 and 0.83,respectively.No significant differences of LAA ostium dimensions (long axis and short axis) and depth were found between the measurement results based on 3D printed models and 3D-TEE (all P＞0.05).A simulation of LAA occlusion rehearsal was successfully performed on 3D models of two challenging cases.Conclusion The echocardiographic 3D printing technique has high feasibility and accuracy,and can be promising for personalized planning in cases of transcatheter special morphological LAA occlusion.
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Objective To improve the canines myocardial infarction curative effect by using ultrasound targeted microbubble destruction (UTMD) combining with nuclear localization signal(NLS) peptide to increase hAng-1 gene transfection efficiency.Methods Forty-six canines were randomly divided into 4 groups (n=10 in each group) after the models of myocardial infarction were prepared.Group A were untreated control;Group B were transfected with hAng-1;Group C were transfected with UTMD+hAng-1;Group D were transfected with UTMD+NLS nuclear localization signal+hAng-1.The therapeutic agents were intravenously injected at one week after myocardial infarction in each group,and the ultrasound were irradiated at the precardium in group C and D.①Echocardiography was used before and at one week after myocardial infarction and 28 days after gene transfection.Two-dimensional echocardiography was used to detect cardiac function,the left ventricular ejection fraction,the left ventricular wall motion and the myocardial contrast echocardiography were used to detect myocardial perfusion of all canines in the four groups.②On twenty-eight days after gene transfection,mRNA and Western Blot were used to detect the expression of hAng-1.Immunohistochemistry was used to detect capillary density of peri-infarct area and microvessel density (MVD).Masson′s trichromatic staining and gross specimen were used to evaluate the degree and the area of myocardial fibrosis.The gene transfection efficiency and the curative effect of all the four groups were evaluated and compared.Results ①Before myocardial infarction,in four groups canine ventricular wall motion and cardiac function were normal,and myocardial filling defect was not showed by myocardial contrast echocardiography.At one week after myocardial infarction,the left ventricular anterior and interval walls motion and the left ventricular ejection fraction in the four groups were significantly decreased.Myocardial contrast echocardiography showed anterior and interval walls myocardial filling defect.There was no significant difference among the four groups(P>0.05);On 28 days after gene transfection the left ventricular ejection fraction in the four group were increased in an order of group A,B,C,D,there was significant difference when comparing group C and D with other groups separately(P<0.05).Myocardial contrast echocardiography showed much contrast filling in the infarction and surrounding area in group D,a little contrast filling in group C and filling defect in group A and group B,there was significant difference when comparing group C and D with other groups separately(P<0.05).②RT-PCR and Western Blot showed the hAng-1 mRNA and protein expression in group D were higher than those in the other group.There was significant difference when comparing group C and group D with other groups separately(P<0.05).③Immunofluorescence showed the capillary densities were(4.7±1.6)/mm2,(11.2±2.8)/mm2,(70.0±6.4)/mm2 and (85.3±7.0)/mm2 in group A,group B,group C and group D.The differences were statistically significant compared group C and group D with other groups (P<0.05).④Masson′s trichromatic staining and cardiac gross specimen showed that the degree and area of myocardial fibrosis were gradually reduced in an order of group A,B,C,D.Conclusions UTMD and NLS peptide could effectively transfect hAng-1 gene and it provided a novel strategy of gene treatment for ischemic heart disease.
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Objective To establish a Chinese hamster model of babesia infection, to find the changing pattern of organs and biochemical parameters in Chinese hamster infected with Babesia, and to promote the detection and treatment of babesiosis.Methods Healthy 5-week old Chinese hamsters were infected by intraperitoneal injection of blood containing Babesia.Blood samples were collected at 0, 2, 4, 6, 8, 10, 12, 14, 16, 23, 30, and 37 days after infection from 5 hamsters at each time point.Blood smears were prepared to detect the parasites using Giemsa staining.ELISA assay was employed to test the IL-2 concentration.The blood biochemical indexes were detected using an automatic biochemical analyzer.DNA was extracted from the whole blood and REAL-TIME RCR was performed to determine the reproduction of Babesia.Aftert the animals were sacrificed, the heart, lung, spleen, liver, and kidney were taken to analyze the changes of organ coefficients.Results The highest level of Babesia in the hamsters occurred on day 4 after the Babesia injection, and then showing a decreasing tendency.However, there was a transient increase on the 12th day after infection.The liver and spleen displayed most extensive response to the infection showing hepatomegaly and splenomegaly, but the variation of heart and kidneys coefficients was within the norm.There were prominent changes of blood cells, especially leucocytes, with two peaks at day 10 and 23 after the Babesia infection.The peak changes of blood biochemical indexes occurred at day 12 after infection.The concentration of serum IL-2 reached a peak on the 10th day after infection.Conclusions The Chinese hamsters display typical characteristics of tick-borne diseases such as hepatomegaly and splenomegaly.The immunological system is activated along with the infection and reaches a highest stage in the second week.Afterwards the Babesia can live in the hamster body for a long period of time.The results of this study provide useful information supporting further studies on the detection, treatment and prevention of Babesiosis.
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Objective To explore the clinical value of two‐dimensional transesophageal echocardiography (2D‐TEE) and real‐time three‐dimensional transesophageal echocardiography (RT3D‐TEE) for the left atrial appendage (LAA) closure procedures by the visualization of LAA shape by 2D‐TEE and RT3D‐TEE and the comparison between the measurement of LAA ostium and the sized LAmbreTM device during the procedure .Methods Forty‐one atrial fibrillation patients ,who had undergone 2D‐TEE examination at our hospital ,were enrolled in the study .At the mid‐esophageal ,dimensions of inner and outer ostium and depth of LAA were measured at the 2D‐TEE views of 0 ,45 ,90 and 135 degree respectively . RT3D‐TEE views were acquired and the maximal and the minimal dimensions of LAA inner ostium were measured .The measurement by RT3D‐TEE and 2D‐TEE were compared to find the difference and correlation .Eleven of 41 patients who have complied with the requirements for the LAA closure ,were undergone the procedures ,measured the dimension of LAA inner ostium at selective angiography intraoperative .Sizes of closure disks of the closure device and the measurement at selective angiography were recorded to compare the measurement at RT3D‐TEE and 2D‐TEE .Results Forty‐one atrial fibrillation patients were completed TEE examination successfully .Inner ostial dimension of LAA was (20.0±04.3)cm,(19.7±03.8)cm,(21.2±04.6)cm,(23.0±05.0)cmat2D‐TEEviewsof0,45,90and135 degree ,respectively .The maximum dimensions of LAA inner ostium by RT3D‐TEE was (2 4.9 ± 0 5.2)cm . At 2D‐TEE views ,the maximum dimensions of LAA inner ostium was at 135 degree ,there was a difference between it and the measurement by RT3D‐TEE ( P =0 0.12) .Monitoring by TEE ,LAA closure procedures with LAmbreTM device were successful for all 11 patients ,the landing zone by selective angiography was (2 4.9 ± 0 4.4)cm ,and the appropriate sized closure disk of the LAmbreTM device was 2 4. - 3 6. cm . Correlation between the measurements by RT3D‐TEE and selective angiography and the sized closure disk were r =0 8.16 ,P =0 0.02 and r =0 9.14 ,P =0 0.00 ,respectively .Correlation between the measurements by 2D‐TEE and selective angiography and the sized closure disk were r =0 6.93 ,P =0 0.18 and r =0 6.88 , P=0 0.19 ,respectively .Conclusions There was better correlation among the measurements by RT3D‐TEE and selective angiography and the size of closure device .Therefore ,compared to 2D‐TEE ,the guidance of RT3D‐TEE was more accurate during LAA closure procedures for LAmbreTM device selection .
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Objective To evaluate the application of real-time three-dimensional transesophageal echocardiography (RT-3D TEE)in left atrial appendage (LAA)closure.Methods After excluding valvular disease,21 of 36 atrial fibrillation (AF)patients suffered the percutaneous LAA closure were chosen to measure left ventricular ejection fraction (LVEF)and to observe if the thrombus or the cloudiness echo will occur.The multiple planner reconstruction function was applied to rebuild sections and to observe the LAA anatomical morphology and its internal structure.During operation,the Flexi Slice function was used to measure the dimensions of LAA ostium,RT-3D mode was used to monitor sheath transport and closure device release,and to immediately evaluate the effect of blocking and complications after the operation. Results The exam before the closure procedure showed 5 of 36 patients' left ventricular ejection fraction (LVEF)<40% and one or more thrombus or the cloudiness echo were found in other 10 patients' which were thus excluded from the study.RT-3D TEE examination in the 21 patients revealed 8 single-lobe cases, 8 double-lobes cases and 5 patients with multi-lobe.Two patients of them need a special device.The maximum dimensions of LAA ostium was (22.24±4.35)mm,the fixed plate size of conventional LAmbreTM device was (28.26±5.23)mm.All patients choiced an average (1 .1 ±0.30)closure device and get the best sealing effect evaluated by RT-3D TEE immediately after operation,10 cases of them has about 1 -3 mm residual leakage exist around the closure.None of the patients with a new pericardial effusion and cardiac tapenade,no closure shift and embolism events.Conclusions RT-3D TEE can real-time and dynamically observe LAA anatomical morphology and its internal structure,compared with other imaging techniques, RT-3D TEE has irreplaceable advantages in the percutaneous LAA closure.
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<p><b>OBJECTIVE</b>To investigate the anti-human CEM lymphoma cell activities induced by TCR idiotypic DNA vaccine containing different antigen determinants in BALB/c mice.</p><p><b>METHODS</b>The specific rearranged gene fragment encoding TCRVbeta region of CEM cell line was obtained by RT-PCR technique. The PCR product was cloned into eukaryocytic expression vector pcDNA3, which was used as DNA vaccine and template for PCR amplifying different antigen determinant. Gene fragments encoding different antigen determinant were amplified and cloned into pcDNA3, separately. The experimental mice were immunized by intramuscular injection of the DNA vaccines. The specific anti-idiotype antibodies were detected by indirect immunofluorescence assay.</p><p><b>RESULTS</b>TCRbetaV of CEM cell line contains five antigen determinants. Specific anti-idiotype antibody was detected in all of the six mice immunized with DNA vaccine containing all the five determinants (the highest titer was 1:480). Although the antibody could also be detected in four of the six mice immunized with DNA vaccine containing four of the five antigen determinants, the antibody titer was lower (the highest titer was 1:80). DNA vaccine containing two of the five determinants could not induce the specific antibody.</p><p><b>CONCLUSION</b>The idiotypic DNA vaccine containing the whole TCRbetaV five antigen determinants could induce the specific anti-lymphoma idiotypic antibody in BALB/c mice.</p>
Subject(s)
Animals , Humans , Mice , Amino Acid Sequence , Antibodies, Anti-Idiotypic , Blood , Allergy and Immunology , Base Sequence , Complementarity Determining Regions , Genetics , Allergy and Immunology , Epitopes , Genetics , Allergy and Immunology , HL-60 Cells , Lymphoma , Allergy and Immunology , Mice, Inbred BALB C , Molecular Sequence Data , Receptors, Antigen, T-Cell, alpha-beta , Genetics , Allergy and Immunology , Sequence Analysis, DNA , Tumor Cells, Cultured , Vaccines, DNA , Genetics , Allergy and ImmunologyABSTRACT
Objective To study the antigenicity of RNA polymerase and the relationship between the protein and the SARS-CoV replication. Methods The fragment was amplified by PCR, ligated with the prokaryotic expression vector pET32a+ and transformed into E. coli BL-21 .The expressed fusion protein identified by Western-blot and ELISA was used to detect the anti-SARS CoV IgG in different sera. Results The fusion protein was expressed successfully in E. coli BL-21. Detected by ELISA, the positive percentage of the anti-SARS IgG in the healthy donors, patients, infected animals and the rhesus administrated with the inactivated-virus were 0%, 95%, 100% and 0%, respectively. Conclusion Good antigenicity was shown in the expressed RNA polymerase. It can be used to diagnose the infection and to demonstrate the replication of SARS-CoV virus.
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This study was undertaken to investigate the anti-lymphocytic malignacy immunologic effects induced by TCR idiotypic DNA vaccine on BALB/c mice. CEM lymphoma cell line and BALB/c mice were used as models. The rearrangement gene fragment coding TCR Vbeta region of CEM cell line was obtained by RT-PCR technique. The PCR product was cloned into the eukaryocytic expression vector pcDNA3 to be used as DNA vaccine. The experimental animals were immunized by intramuscular injection with DNA vaccine. The specific anti-idiotypic antibody was detected by indirect immunofluorescence assay. The specific anti-idiotypic cellular immunity was detected by CTL activity assay using MTT method. The results showed that specific anti-idiotypic antibody in the immunized mice sera could be found since four weeks after immunization and came to the peak of titer on the sixth week. Using IL-6 as immunological adjuvant could significantly increase the antibody titers. It was concluded that the TCR idiotypic DNA vaccine could induce effectively the specific anti-lymphoma idiotypic antibody in BALB/c mice. Using IL-6 as immunological adjuvant could significantly increase the antibody titers induced by idiotipic DNA vaccine.