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Objective:To investigate the effects of NOD-like receptor protein 3(NLRP3) inflammasome activation on the proliferation, migration and extracellular matrix desposition of activated pancreatic stellate cells(PSCs).Methods:The rat PSCs were isolated, cultured and identified, and were divided into control group or LPS group based on the pretreatment with LPS (10 μg/ml for 24 hours) or without. The expression of NLRP3 inflammasome associated molecules in PSCs culture medium was detected by ELISA. The PSCs with NLRP3 inhibition were constructed by shRNA carrying lentivirus infection and were divided into LPS+ negative control group and LPS+ lentivirus group based on whether the cells were treated with LPS and infected by lentivirus or not. The alteration in cell proliferation and migration were detected by CCK-8 kit and transwell chamber method. The expression of extracellular matrix α-SMA and collagen in PSCs was detected by immunofluorescence staining and the expression of TGF-β mRNA was analyzed by RT-qPCR.Results:The cytoplasm of PSCs which were cultured for 24 hours was rich in bright annular lipid droplets, and the cells expressed desmin. After 7 days of culture, the cell became larger in size, the lipid droplets basically disappeared, and the cells were activated and expressed α-SMA. The expression of caspase-1, IL-1β and IL-18 in the supernatant of PSCs culture medium in LPS group were significantly higher than those in control group (1.55±0.04 vs 0.65±0.03), (2.02±0.04 vs 1.05±0.05) and (1.70±0.05 vs 0.97±0.03), respectively. After inhibiting by lentivirus infection, the expression of NLRP3 in the lentivirus group (0.25±0.04) was significantly lower than that in negative control group (0.68±0.05). In control group, LPS group, LPS+ negative control group and LPS+ lentivirus group, the A490 values was 0.61±0.02, 1.15±0.06, 0.96±0.05, and 0.56±0.01, respectively; the migrating PSCs number was (64.12±4.58), (121.67±8.02), (111.67±4.67) and (69.67±8.08)/HF, respectively; the relative expression of α-SMA was 0.78±0.05, 4.12±0.04, 3.81±0.06 and 0.88±0.05, respectively; the relative expression of collagen was 0.65±0.03, 3.43±0.02, 2.67±0.02 and 0.48±0.03, respectively; and the expression of TGF-β mRNA was 0.22±0.03, 0.89±0.01, 0.86±0.03 and 0.43±0.02, respectively. The A490 value, the migrating cells number, the expression of α-SMA, collagen and the expression of TGF-β mRNA in LPS group and LPS+ negative control group was significantly higher than those in control group and LPS+ lentivirus group, and all the differences were statistically significant (all P value <0.05). Conclusions:NLRP3 inflammasome activation may accelerate the extracellular matrix deposition and pancreatic fibrogenesis by promoting PSCs proliferation and migration ability via regulating the biological functions.
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Objective:To investigate the application value of CT and MRI imageomics based on machine learning method in the diagnosis of pancreatic cancer.Methods:The clinical data of 62 patients with surgically resected and pathologically confirmed pancreatic cancer, who underwent enhanced CT scan, MRI plain or enhanced scan in Shanghai General Hospital between January 2014 and December 2021 were collected. According to the chronological order of surgery, 49 patients from January 2014 to December 2020 were enrolled in the training set and 13 patients from January 2021 to December 2021 were enrolled in the validation set. 3D-slicer 4.8.1 software was used to draw the region of interest in each layer of CT and MRI images for cancerous and paracancerous tissue segment. Image features were extracted by Python and the optimal feature set from the training set data was obtained by using Lasso regression model. The machine learning decision tree model was constructed. The receiver operating characteristic curve(ROC) curve was drawn, and the area under the curve (AUC) was calculated to evaluate the value of these three kinds of imageomics models in the diagnosis of pancreatic cancer.Results:The 1 767 CT features and 1 674 MRI features were obtained from enhanced CT scan, MRI plain scan and enhanced MRI scan, respectively. For the differential diagnosis model of cancerous tissue and paracancerous tissue, the enhanced CT scan data model obtained the optimal feature set involving 6 features, the MRI plain scan model obtained the optimal feature set involving 16 features, and the enhanced MRI scan model obtained the optimal feature set involving 15 features. The diagnostic model based on enhanced CT scan had an AUC of 0.98 in the training set and 1 in the verification group. The AUC of the MRI plain scan and enhanced MRI scan models in both the training set and the validation set was 1. The specificity and sensitivity of machine learning decision tree model based on the three kinds of imageomics models in the diagnosis of cancerous tissue and paracancerous tissue were 100%. For the differential diagnosis model of splenic artery wrapping, the enhanced CT scan model didn′t obtain the optimal features and had no diagnostic efficacy. The MRI plain scan model and enhanced MRI scan model obtained the optimal feature set involving 5 and 4 features, respectively. The AUC of the MRI plain scan model in the training set and the validation set were 0.862 and 0.750, respectively, with diagnostic sensitivity of 93.8% and 50.0%, and specificity of 78.6% and 100%, respectively. The AUC of the enhanced MRI scan model in the training set and the validation set were 0.950 and 0.861, respectively, with diagnostic sensitivity of 90.0% and 93.6%, and specificity of 100% and 78.6%, respectively.Conclusions:Based on the radiomics of CT enhanced, MRI plain scan and enhanced MRI scan, the machine learning diagnostic model has an accuracy of more than 90% in differentiating pancreatic cancer from paracancerous tissue. For the differentiation of splenic artery wrapping in pancreatic cancer, the diagnostic model based on enhanced MRI scan haS the best diagnostic efficiency.
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Objective To investigate the clinical value and surgical methods of pancreaticoduodenectomy (PD) combined with portal vein (PV)/superior mesenteric vein (SMV) resection and reconstruction in the treatment of pancreatic cancer with PV/SMV invaded by tumor.Methods The clinical data of 21 patients of pancreatic cancer with PV/SMV invaded by tumor (group A) and 62 patients of pancreatic head cancer without PV/SMV invaded by tumor (group B) in the same period were collected and analyzed retrospectively from Jan 2014 to Apr 2017.There were no distinct invasion of celiac artery (CA),hepatic common artery (HCA) and superior mesenteric artery (SMA) in two groups of pancreatic cancer patients.The patients of group A underwent PD combined with PV/SMV resection and reconstruction,and the patients of group B were only treated with PD surgery.The complication rate and overall survival time after PD was compared between the 21 patients of pancreatic cancer with PV/SMV invaded by tumor and the 62 patients of pancreatichead cancer without PV/SMV invaded by tumor.'Results The average overall survival time of 21 patients of pancreatic cancer with PV/SMV invaded by tumor (group A) was 19.2 months,specifically with 1-year survival rate of 57.1% (12/21),2-year survival rate of 28.6% (6/21),and 3-year survival rate of 14.3% (3/21).Meanwhile,the average overall survival time of group B was 19.4 months,specifically with 1-year survival rate of 58.1% (36/62),2-year survival rate of 30.6% (19/62),and 3-year survival rate of 14.5% (9/62).The results indicated that no differences for overall survival time of patients treated with PD including 1,2,3-year survival rate between two groups were found (P > 0.05).Conclusions For pancreatic cancer accompanied by PV/SMV invasion without invasion of SMA,CA and HCA,PD combined with PV/SMV resection and reconstruction are safe and feasible surgical procedures.The surgical reconstruction method was determined according to the location and length of the invaded vessels,and also there were no significant differences on the complication rate and overall survival time after PD between the pancreatic cancer patients with invasion of PV/SMV and the pancreatic head cancer patients without invasion of PV/SMV.
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Objective The clinical benefits of carbohydrate intake 2-3 h before surgery in patients with digestive tract malignant tumor without diabetes mellitus have been confirmed by foreign experts,the"High level evidence" for perioperative nutrition support were also written in China"Guidelines for perioperative nutritional support in adults".However,there are few hospitals in China that prefer preoperative glucose pretreatment,taking into account the differences between domestic and foreign data.In this paper,the clinical randomly controlled experimental data about the effect of preoperative glucose pretreatment on the prognosis of patients with malignant tumor of digestive tract was analyzed by Meta analysis,in order to evaluate the efficacy and safety of preoperative glucose pretreatment in Chinese patients,and to achieve the clinical significance.Methods Retrieving the database from establishment to March 2017,the related literatures about preoperative glucose pretreatment and prognosis of patients with digestive tract malignant tumor were collected.Revman5.3 software was used to perform Meta analysis of 10 randomized controlled trials(606 patients)who met the inclusion criteria.Results Compared with the traditional fasting and non drinking group,the preoperative subjective discomfort of the preoperative group was significantly reduced(SMD = -1.29,95%CI = -2.00--0.59,P = 0.000 3), postoperative insulin resistance was significantly reduced(SMD=-1.92,95%CI=-2.68--1.16,P<0.01), postoperative blood glucose concentration,insulin concentration increased,and no vomiting and aspiration occur in any patient as shown by the literature.Conclusion In the patients with malignant tumor of digestive tract, perioperative nutrition support and pretreatment show significant clinical benefits,preoperative subjective comfort improves,the risk of aspiration does not increase significantly,postoperative insulin resistance decreases.It is proved that preoperative carbohydrate pretreatment is effective and safe in the operation of domestic digestive tract cancer.
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Objective To prepare a targeted ultrasound micro bubble,which carried the HSV-TK gene,and investigate the in vitro target searching ability of the micro bubbles and inhibitory effect on the proliferation of HepG2 cells.Methods Ultrasonic micro bubbles were prepared by mechanical vibration method,construction of targeted HSV-TK ultrasound micro bubbles by biotin affinity bridge construction.To detect the general characteristics of ultrasound micro bubbles,and to test its effect on the proliferation of HepG2 cells in vitro.Results HSV-TK targeted ultrasound microbubbles more gathered on the surface of HepG2 cells,through detection of PCNA and MTT,it was found that the proliferation of gene targeting microbubble group was obviously decreased,cell apoptosis increased significantly,Cells invade experiments showed that the number of cells in genetic microbubble group (22.18 ± 2.01) decreased significantly compared with the control group and the nontargeted group,can effectively inhibit the proliferation and invasive ability of HepG2 cells.Conclusion Targeted ultrasound microbubble carrying target gene have better inhibitory effect on HepG2 cells in vitro.
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Objective: To explore the echocardiographic cardiac geometric morphology and hemodynamics in premature infants at different gestational age with the inlfuencing factors. Methods: A total of 150 premature infants and 150 full-term control infants were enrolled in this study. Based on gestational age, premature infants were divided into 3 groups:①(28-32+6 ) weeks,②(33-34+6 ) weeks,③(35-36+6) weeks; and full term control infants were divided into 2 groups:①’(37-38+6) weeks and②’ (39-41+6) weeks respectively. An iE33 Philips ultrasound examination was conducted to measure left ventricular end-diastolic diameter (LVEDD), LVESD, interventricular septum thickness, posterior wall thickness, left ventricular end-diastolic volume (LVEDV), LVESV, stroke volume, LVEF, left ventricular fractional shortening (LVFS), cardiac output, stroke index, cardiac index, left ventricular mass, left ventricular mass index (LVMI), left ventricular relative wall thickness, left ventricular remodeling index (LVRI) and LVEDVI. Results: With adjusted body surface area, all parameters for cardiac geometric morphology and hemodynamics were similar among different groups,P>0.05. The day-old age (P=0.001), height (P=0.001) and body weight for low weight born infant (P=0.012), for normal weight born infant (P=0.003), for giant infant (P=0.016) were the independent inlfuencing factors for LVMI. The impact of anthropometry and the basic life indexes were similar on LVRI among groups (χ2=42.88,P=0.076), while the covariates were different on LVMI among groups (χ2=123.6,P Conclusion: Cardiac morphology and hemodynamics measured by echocardiography has important clinical meaning for assessing the development and maturity of neonatal hearts in premature infants.
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Objective To observe the effect of glutamine-enriched early enteral nutrition on intestinal mucosal barrier injury after orthotopic liver transplantation in rats.Method Male Wistar rats,the recipients were randomly divided into three groups:control group (control group,n =10),orthotopic liver transplantation group (OLT group,n =30) and glutamine-enriched early enteral nutrition group (EEN group,n =30).Only dissecting hepatoduodenal ligament was performed in control group,and OLT was performed from Wistar to Wistar rats by modified two-cuff method in OLT group and EEN group.For EEN group,recipients were supplied with Nutrison Fiber (125 ml/kg every day) plus Gln (0.3 g/kg every day) for 3 days before and 3 h after surgery by gastric perfusion.For OLT group,the same volume of normal saline was administered instead of the Nutrison in the same time.No special treatment was given in control group.The levels of plasma endotoxin,D-lactic acid,and TNF-α were determined at different time points in the three groups.The ultrastructural changes of ileal mucosa were observed under the transmission electron microscopy.At the same time,the remaining 5 rats per group were used for observing the lifetime.Result As compared with control group at 12,24 and 72 h,the levels of plasma endotoxin,D-lactic acid,and TNF-α were significantly increased in OLT group and EEN group (P<0.01),and as compared with OLT group,the above-mentioned indexes were obviously decreased in EEN group at 24 and 72 h (P< 0.01).The ileal mucosal epithelial clearance in control was normal,and microvilli arranged neatly under the electron microscope.the ultrastructure damage in OLT group was more serious than in EEN group at 12,24,and 72 h.As compared with control group at 12,24,and 72 h,the expression levels of TNF-α mRNA were significantly increased in OLT group and EEN group (P<0.01),and as compared with OLT group,the expression levels of TNF-α mRNA were obviously decreased in EEN group (P<0.01).The survival time in cntrol group was significantly longer than in OLT group and EEN group (P<0.05),and as compared with OLT group,the survival time in EEN group was obviously extended (P<0.05).Conclusion OLT can lead to the damage of intestinal mucosal barrier,and glutamine-enriched early enteral nutrition is a potent protection against intestinal mucosal barrier injury and prolongs the survival time of rats after OLT.
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Objective To summarize the technique and clinical experience of hybrid procedure under cardiopulmonary bypass (CPB) in children with muscular ventricular septal defect (mVSD). Methods From January 2006 to June 2010, 45 cases of mVSD underwent hybrid procedure with CPB. mVSDs were closed with devices under direct vision in 45 cases. Of them, there were 20 males and 25 females. They ranged from 52 days to 12 years [mean (2.05 ±2.48) year] in age and from 3 to 30 kg [(11.93 ±7.70)kg] in body weight. Preoperatively, most of children were highly susceptible to respiratory tract infections. The hybrid approach was used in all patients with CPB under the guidance of transesophageal echocardiography (TEE). The diameter of mVSDs ranged from 2 to 7 mm under TEE. Of 45 cases, 40 patients had increased rates of pulmonary blood flow. 29 patients had left axis deviation and 12 cases had sinus arrhythmia on electrocardiography (ECG). 19 had other congenital heart lesions, including transposition of great arteries in 1 case, tetralogy of Fallot in 2, pulmonary artery stenosis in 3, patent ductus arteriosus in 6, atrial septal defects in 6) and aorta coactation in 1. The quantity of VSDs were from 1 to 7 (single, in 7; two, in 24 case; three, in 8 case; four, in 5 case and seven, in lease. 37 patients were combined with pulmonary hypertension in our cohort. 38 patients with another large VSD and 19 with other congenital heart lesions were required surgical repair at sometime. Results The hybrid procedures were undertaken in all 45 cases of this cohort. All cases were successful and no deaths occurred during operation. A total of 48 devices were implanted in 45 patients, including single devices in 42 cases (device size ranged from 4 to 10 mm) and two devices in 3 cases (device size ranged from 4 -7 mm). The average time on CPB was (58.28 ±20.70) min , while aortic crossclamp time was(34. 94 ± 14.75) min. In addition, the time on mechanical ventilation postoperatively ranged from 2 hours to 6 days. Compared to the older children, 20 infante aged less than 8 monhad a significant difference in cardiac function in the early postoperative period. One infant was given up treatment because of serious infection. Anather cases recovered with the use of supportive treatment, such as using vasoactive agents, digoxin, inhaling nitric oxide, diuresis, and so on. The enteric-coated aspirin was given at dose of 5 mg ? kg -1. day -1 for a period of 3 to 6 months as usual postoperatively. All patients attended follow-up at 1 week, 1 month, 3 months, 6 months, 1 year and 2 years post-procedure. No major complications were encountered during this period. All cases were no instance of migration of any of the devices, residual shunt, aortic regurgitation, atrioventricular valve dysfunction, Ⅲo atrial-ventricular conduction block, new arrhythmia, and so on. There are no death in long-term follow-up. Conclusion Hybrid procedure is safe and effective for the closure of congenital heart defects in children.
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Objective To review the clinical experiences of modified Nuss procedure and technique for complicated pectus excavatum. Methods From September 2006 to October 2009,45 children (27 boys and18 girls), aged 3 - 16 years [mean (6.4 ± 3.5 )years] with complicated pectus excavatum underwent modified Nuss procedure. VATS was used in 6 cases. 3 patients associated with lung cyst were treated by open surgery and 1 case with ASD was treated by Hybrid procedure.Results The surgery was successfully performed in all patients. The mean operative time was 54 minutes and the average hospital stay was 7 days. Postoperative evaluation was"excellent"in 30 cases , "good"in 13, and"moderate"in 2. Conclusion Modified Nuss procedure is a safe and effective method for correction of complicated pectus excavatum.
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Objective To investigate the changes of cerebral blood flow and the level of brain injury in a rat model of deep hypothermia low flow(DHLF).Methods Twelve SD rats aged 3 weeks were randomly divided into sham group and model group.Regional cerebral blood flow(rCBF)of all rats was measured continuously during the operation by laser Doppler flowmetry,and the changes of rCBF were measured before temperature decreased,when the temperature decreased to(21.0±0.5)℃,0-5 minutes,25-30 minutes,55-60 minutes,115-120 minutes during DHLF and 0-5 minutes after DHLF operation.Another 60 SD rats aged 3 weeks were randomly divided into sham group and model group.Six rats of each group were sacrificed at 1,6,24,72 hours and 7 days after DHLF operation to detect the pathological changes of the brain and the neuronal apoptosis by HE staining and TUNEL assay.The neurological deficit score(NDS)was recorded at 24,72 hours and 7 days after operation for evaluating the neurologic functional outcome.ResultsWhen the temperature was decreased to(21.0±0.5)℃,the levels of rCBF of sham group and model group were significantly decreased to(41.1±4.2)% and(40.7±3.4)% of the baseline value,and the rCBF level of model group was further decreased to(15.7±3.5)% of the baseline value 0-15 minutes during DHLF(P < 0.01),with no obvious changes in all the time intervals during DHLF.Compared with the sham group,the scores of NDS of model group were significant lower at 24 and 72 hours after operation(P < 0.05 or 0.01).Besides,a significant pathological change of the brain tissue and a increased percentage of TUNEL-positive staining cells were observed in model group at 6,24,72 hours and 7 days after operation(P < 0.01).Conclusions Rat model of DHLF is an ideal and reliable model of brain injury,for it is similar to DHLF procedure of clinical cardiac operation.
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Ovbective To evaluate the outcome of the complete repair for critical and complex congenital heart defects in small infants with cardiopulmonary bypass. Methods A retrospective review of hospital records was performed for 187 infants under 6 months undergone cardiac operation from January 2000 to January 2004. Average age was (4.15?2.32) months (3d-6months) and average weight was (4.42?0.56)kg (3~7kg). The diagnoses included ventricular septal defect with pulmonary hypertension,transposition of the great arteries,aortic coarctation with other lesions,total anomalous pulmonary venous connection,tetralogy of Fallot,atrioventricular canal defect,pulmonary atresia,and neonatal cardiac tumor. Average duration of mechanical ventilation was (17.5?9.7) hours. ICU stay was (4.9?1.5) days. Results 9 cases died after operation with a hospital mortality of 4.81%. Postoperative complications included low cardiac output syndrome,residual shunt,pulmonary infection and pulmonary hemorrhage. 178 are alive and in good cardiac status at follow-up of 4~48 months. After repair,growth approximates the normal. Conclusion With the development of pediatric cardiac surgery, cardiac operation with CPB in small infants with critical and complex congenital heart defects is feasible and may provide satisfactory survival.
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Objective:To study the effect of RNA interference(RNA)ion ATX-mRNA expression and the invasive petential of human breast cancer cell lines.Methods:Chemically synthesizeddouble stranded RNA(dsRNA)targeting ATX was transfected into human breast cancer cell MCF-7 using SiPORT Lipid.The transfection efficiency was observed under fluorescence confocal microscope.Expression of ATX mRNA and protein was detected by reverse transcription polymerase chain reaction(RT-PCR)and Western blot.Cell penetrate matrigel capacity were determined by in vitro experiment.Results:ATX-siRNA effectively inhibited ATX-mRNA and protein expression(P
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Objective To evaluate the effects of ischemic preconditioning(IP) on liver function,complications and hospital stays after hepatectomy under hepatic vascular exclusion by a meta-analysis.Methods Randomized controlled trials(RCTs) were identified from PUBMED,EMBASE,the Cochrane Library,VIP,CNKI and Wanfang Data according to the inclusion and exclusion criteria.Literature screening,data extraction and quality assessment were made and the meta-analysis was processed by RevMan 4.2.2.Results Eight RCTs involving a total of 511 patients were included.The methodological quality was evaluated and all the trials were in graded B.The meta-analysis revealed that the postoperative ALT peak level(weighted mean difference=-176.37;95%CI:-320.67~-30.06;P=0.02)and postoperative complications incidence(odd ratio=0.64;95%CI: 0.41~0.98;P=0.04)were lower in IP group compared with control group,but there were no significant differences in blood loss,operating time,hepatic vascular exclusion time,postoperative AST and total bilirubin peak level,and hospital stays in both groups.Conclusions IP reduces the postoperative ALT peak level and complications incidence after hepatectomy under hepatic vascular exclusion,but there is no sufficient evidence to support that the IP can protect the liver from ischemia/reperfusion injury.