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Objective:To explore the application of laparoscopic training in the standardized residency training of pediatric general surgery.Methods:A total of 36 rotating residents who received the standardized residency training in the department of pediatric general surgery of Shenzhen Children's Hospital from January 2017 to December 2019 were selected for laparoscopic training. The training content includes the study and training of laparoscopic theory knowledge, laparoscopic surgery video, skills operation of simulated operating system. Assessment was conducted before and after the training, and statistical analysis was performed to compare the difference of scores before and after the training. The self-evaluation of resident's learning efficiency and the satisfaction with teachers were investigated by questionnaires. SPSS 20.0 was used for paired t-test. Results:After receiving the standardized residency training of pediatric general surgery, the theoretical knowledge and cognition of the application on pediatric general laparoscopic surgery were significantly improved among the 36 residents. The time that the skills spent in vitro simulation box during the simulated operation training was significantly reduced after training, with a statistically significant difference ( P < 0.05). According to the questionnaire survey, resident's satisfaction with self-evaluation of learning efficiency was 97.22% (35/36), their satisfaction with teachers was 94.44% (34/36), and their satisfaction with teaching curriculum was 100.00% (36/36). Conclusion:Laparoscopic training can effectively improve the clinical practice ability of pediatric general surgery residents, which is worthy of promotion.
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Objective To investigate the mechanism of miRNA-129-5p in epithelial-mesenchymal transition (EMT)of biliary atresia.Methods Constructed bile duct epithelial EMT cell model (the experimental group) induced by TGF-β1,detected the expressions of EMT related markers and miRNA-129-5p.While miRNA-129-5p precursor was transfected,the expressions of EMT related markers and extracellular matrix were contrasted between the original and the renovated biliary epithelial cells.Results In the experimental group,extrahepatic bile duct showedEMT,the expression of miRNA-129-5p was decreased (P＜0.05),overexpression of miRNA-129-5p could inhibit the progression of EMT (P＜0.05).Conclusion miRNA-129-5p may relate to EMT by regulating the expression of TGF-β1.
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The aim of this study is to develop creatine kinase isoenzyme MB (CK-MB) specific monoclonal antibodies (mAb), and characterize the monoclonal antibody and further development of quantitative detection assay for CK-MB. The BALB/c mice were immunized with purchased CK-MB antigen, then monoclonal antibodies were prepared according to conventional hybridoma technique and screened by indirect and capture ELISA method. To identify the epitopes and evaluate the classification, purchased creatine kinase isoenzyme MB (CK-MM/BB/MB) antigen was used to identify the epitopes, with immunoblotting and synthetic CK-MM and CK-BB in different linear epitope. A double antibody sandwich ELISA was applied to screen the mAb pairs for CK-MB detection, and the quantitative detection assay for CK-MB was developed. We used 74 cases of clinical specimens for comparison of our assay with Roche's CK-MB assay. We successfully developed 22 strains of hybridoms against CK-MB, these mAbs can be divided into linear, partial conformational CK-MB, CK-MM or CK-BB cross monoclonal antibody and CK-MB specific reaction with partial conformational monoclonal antibody, and CK-MB quantitative detection assay was developed by using partial conformational monoclonal antibody. The correlation coefficient factor r of our reagent and Roche's was 0.930 9. This study established a screening method for CK-MB partial conformational specific monoclonal antibody, and these monoclonal antibodies were analyzed and an established quantitative detection assay was developed. The new assay had a high concordance with Roche's.
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The aim of this study is to prepare and characterize cardiac troponin T (cTnT) monoclonal antibodies (mAb), and further develop a chemiluminescence quantitative detection assay for cTnT. BALB/c mice were immunized with recombinant cTnT antigen, and specific mAbs were prepared using conventional hybridoma technique and screened by indirect ELISA method. To identify the epitopes, several cTnT peptide fragments were synthesized or expressed by genetic engineering. A double antibody sandwich ELISA method was used to screen the mAb pairs for cTnT detection, and the automatic chemiluminescence detection assay for cTnT was developed. In total 220 clinical specimens were used for system comparison between our assay and Roche cTnT assay; further performance characteristics was evaluated by testing 238 clinical samples and 784 physical examination samples. We successfully screened 33 strains of hybridoms against cTnT, and the mAbs' epitopes were identified. Mab E16H8 and C8G11 with a detection limit of 10 pg/mL cTnT antigen were selected to develop the full automatic chemiluminescence quantitative assay. The correlation coefficient of our reagent with Roche's was 0.959 9, with a coincidence rate of 95%. The assay presented a sensitivity of 97.5%, and a specificity of 99.15% in detection of clinical samples. The cTnT concentration was less than 0.080 6 ng/mL in 99% of general population, which agrees with the definition of WHO on patients with acute myocardial infarction (AMI). In summary, we developed monoclonal antibodies against predominant epitopes for diagnostics of cTnT, and an automatic tubular chemiluminescence quantitative detection assay was further developed, which presents a high coincidence rate with Roche's.
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Humans , Antibodies, Monoclonal , Allergy and Immunology , Enzyme-Linked Immunosorbent Assay , Epitopes , Allergy and Immunology , Hybridomas , Luminescent Measurements , Myocardial Infarction , Peptide Fragments , Sensitivity and Specificity , Troponin T , Allergy and ImmunologyABSTRACT
Objective To investigate the relationship between heat shock protein 72 (HSP72) and liver fibrosis in pediatric patients with biliary atresia and the significance of its expression.Methods The clinical and pathological data of 30 patients with biliary atresia were selected from June 2013 to June 2015,and the expression of HSP72 in liver tissues was determined by streptomycin peroxide (SP) immunohistochemical method.Results The greater group of liver fibrosis degree with biliary atresia was less than the lighter group in the expression of HSP72 (Z =-4.16,P ＜ 0.01).There was negative correlation between HSP72 expression intensity and operation age (r =-0.704,P ＜ 0.01).Meanwhile,there was negative correlation between HSP72 expression intensity and postoperative cholangitis (r =-0.371,P ＜ 0.05).When the expression of HSP72 was higher,the autologous liver survival rate was higher (x2 =9.482,P ＜ 0.01).Conclusions The expression of HSP72 in liver tissues of patients with biliary atresia is closely related to the degree of liver tissue fibrosis.HSP72 expression may be one of the factors affecting the prognosis of patients with biliary atresia,and can be used as a reference index for evaluating prognosis.
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Objective To investigate the effect of cold autologous blood cardioplegia and HTK solution on changes of superoxide dismutase (SOD) and Matrix metalloproteinase-2 (MMP-2) in infant coronary sinus vein blood,which underwent ventricular septal defect suture in extracorporeal circulation,in order to reveal their protective effect of autologous cold blood cardioplegia on immature myocardium.Methods Sixty cases with ventricular septal defect,aged less 1-year-old,were randomly divided into experimental group (n =30) and control group(n =30).Autologous cold blood cardioplegia was obtained on aortic blood before aortic root draw in extracorporeal circulation with K + concentration of 20 mmol/L and 4 ℃ temperature.The experimental group was used cold autologous blood cardioplegia as arresting and protecting cardioplegia,while the control group used HTK solution.Blood sample from coronary sinus and vein was obtained at immediately before aortic cross-clamping,1 minute and 15 minutes after aortic cross-opening.SOD and MMP-2 levels were determined by enzyme-Linked Immuno Sorbent Assay (ELISA).Results The levels of SOD and MMP-2 levels within the group,between the groups and interaction were significant difference (P ＜ 0.05).After aortic cross-opening,the level of SOD and MMP-2 of the experimental group and the control group were increased (P ＜ 0.05).Before aortic cross-clamping,The activity of SOD in experiment was(85.37 ± 16.82) mU/L,as same as that in the control group((91.51 ± 15.02) mU/L,P ＞ 0.05).The MMP-2 concentration was in experiment group was (362.29 ±29.52) μg/L,as same as that in the control group((372.32 ± 31.42) μg/L,P ＞ 0.05).At 1 minute after aortic cross-opening,there were significant differences regarding of SOD and MMP-2 levels between the both groups(SOD ∶ (106.97 ± 17.46) mU/L vs.(98.74 12.54) mU/L,P ＜ 0.05 ; MMP-2 ∶ (439.48 ± 51.62) μg/L vs.(465.49 ±48.83) μg/L,P ＜0.05) ;The same trend was seen at 15 minutes after aortic cross-opening in two groups in terms of SOD and MMP-2 level (SOD:(104.03 ± 12.63) mU/L vs.(97.94 10.87) mU/L,P ＜0.05; MMP-2:(390.16 ±45.63) μg/L vs.(425.21 ±48.24) μg/L,P ＜0.05).Compare to group B,arrhythmia incidence(x2 =8.223,P ＜ 0.05) and positive inotropic drug dependent degree was lower in group A (x2 =4.022,P ＜ 0.05).Conclusion The cold autologous blood cardioplegia could promote the release of SOD in immature myocardium and reduce the production of MMP-2,which has endogenous protective effect for the infant Imature myocardium.
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Objective To investigate the impact of leukocyte-removed priming fluid on plasma inflammatory factors TNF-?,IL-6,IL-8,NE peri-CPB in infants.Methods Selected 60 infants,of less than 1-year-old,with ventricular septal defect(VSD)were randomly assigned to the experimental group and the control group.The experimental group took blood leukocyte-removed priming fluid,and the control group took banked blood-derived priming fluid.The plasma TNF-?,IL-6,IL-8,NE concentrations were determinated by using ELISA at the corresponding period of cardiopulmonary bypass(before CPB,aortic cross-clamping 10 min,aortic open 10 min,2h,6h,12h,24h and 48 h after the end of CPB).Results TNF-?,IL-6,IL-8,NE concentrations increased significantly after the start of CPB(P
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Objective To investigate the impact of leukocyte-removed priming fluid on lung function peri-CPB in infants. Methods Selected 60 infants of less than 1-year-old, with ventricular septal defect (VSD), were randomly assigned to the experimental group and the control group, the experimental group took blood leukocyte-removed priming fluid, and the control group took banked blood-derived priming fluid. Indicators of OI, PaO2/PAO2, A-aDO2 and RI were recorded at the corresponding period of cardiopulmonary bypass (pre-CPB, 2h, 6h, 12h, 24h and 48h post-CPB). Results At 2h, 6h,12h post-CPB, OI and PaO2/PAO2 of the experimental group were higher than those of the control group (P
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In order to set up sustainable fundraising mechanisms for community health services, it is necessary to calculate the sum of compensation needed for basic public health services and formulate compensation policies on the basis of defining items of basic public health services and in accordance with the economic levels of various regions. At the same time, it is necessary to enforce government purchases after working out strict assessment criteria so as to ensure that funds for community health services are genuinely devoted to basic public health services.