Objective To discuss features of laparosco pi c cholecystectomy (LC) in children. Methods A total of 54 chi ldren underwent LC from October 1992 to December 2003. The operations were carri ed out after the creation of a CO2 pneumoperitoneum at the flow rate

Objective:To explore the lactate metabolism in brain tissue of the mice with early acute hypoxia-ischemia injury,and to provide data support for 9.4T 1 H-NMR spectroscopy in detecting the lactate level clinically.Methods:Eighty Kunming mice were randomly divided into sixteen groups (0 s,20 s,40 s,60 s,2 min,4 min, 6 min,8 min, 10 min, 12 min, 14 min, 16 min, 18 min,and 20 min)according to the duration of hypoxia-ischemia (n=5).The changes of lactate levels were detected by 9.4T 1 H-NMR spectroscopy. Results:After the initiation of hypoxia-ischemia injury,the lactate level began to increase rapidly to the highest value of (6.89 ± 0.34)μmol·g-1 at 20 s,then started to decline quickly from 40 s to 2 min,and eventually decreased to a stable level of (4.85±0.36)μmol·g-1 until 6 min.Compared with control group,the levels of lactate in brain tissue of the mice in hypoxic-ischemic groups were increased (P <0.01).Conclusion:40 s of acute hypoxia-ischemia may be the lactate cerebral neuron threshold during the anaerobic glycolysis. 9.4T1 H-MRS can provide the exact time window for detecting the lactate metabolism.

Objective To investigate the relationship between the plasma IL-6,ACE level and DVH parameters radiation pneumonitis,and to evaluate its clinical value on the prediction of radiation pneumonitis.Methods 60 patients with locally advanced non-small cell lung cancer patients with unresectable or unwilling to surgical resection routinely received three-dimensional conformal radiation therapy.Before radiotherapy,radiotherapy beginning 1 week,2 weeks,3 weeks,4 weeks,5 weeks,6 weeks,8 weeks and 12 weeks,enzyme-linked immunosorbent assay (ELISA) was used to detect IL-6 and ACE content.The evaluation of radiation pneumonitis was accordance with the RTOG acute radiation pneumonitis grading standards.The patients with radiation pneumonitis were included in observation group,otherwise as control group.IL-6,ACE and DVH parameters in two groups were analyzed.The DVH parameters includes V20,V20,MLD and NTP.Results Among all the 60 patients,16 cases were diagnosed as radiation pneumonitis,among whom 5 cases occurred in radiotherapy,10 cases occurred within one month after radiotherapy,1 case occurred in two months after radiotherapy,including the 11 cases of grade Ⅱ and the other 5 cases of grade Ⅲ.In the observation group and the control group,expression level of the IL-6 had statistical significance before and after radiotherapy,ACE had significant difference in 2rd,3rd,4th,5th,6th week after radiotherapy (all P ＜ 0.05).V20,V30 and MLD differences were statistically significant between the observation group and control group [(46.2±4.5) ％ vs (30.5±7.5) ％,(37.5±5.6) ％ vs (20.5±5.6) ％,(20.4±2.3) Gy vs (15.5±3.5) Gy,(25.2±8.2) ％ vs (9.9±4.5) ％,all P ＜ 0.05].Conclusion Plasma IL-6,ACE levels,V20,V30,the MLD,and NTP are related to the occurrence of radiation pneumonitis.

BACKGROUND:Human-mammal chimeric liver chimera has been a vital significance for the proliferation and differentiation of bone marrow mesenchymal stem cells.
OBJECTIVE:To establish an animal model of human-rhesus chimeric liver using adult bone marrow mesenchymal stem cells.
METHODS:Adult bone marrow mesenchymal stem cells were isolated, purified and cultured for the sixth generation. The number of bone marrow mesenchymal stem cells was no less than 5×108. Bone marrow mesenchymal stem cells labeled with green fluorescent protein were transplanted into the liver of the embryo rhesus with pregnancy of 10 weeks under guided by type-B ultrasound. At the 1st and 3rd months of birth, the liver tissue of the infant rhesus was taken for biopsy. After routine pathological section, histological specimens were observed under fluorescence microscope to confirm if there were adult bone marrow mesenchymal stem cells positive for green fluorescent protein and their distribution, and detected by immunohistochemical staining to identify if human albumin expressed in the liver of infant rhesus.
RESULTS AND CONCLUSION:Fluorescence microscope observation indicated that at the 1st and 3rd months after birth, there were surviving bone marrow mesenchymal stem cells derived from human with green fluorescence in the liver of infant rhesus, and these cells migrated to form more concentrated distribution. The immunohistochemical results demonstrated that functional liver cells expressing human albumin were observed in the liver of infant rhesus at the 1st and 3rd months after birth, and their distribution was in accordance with bone marrow mesenchymal stem cells with green fluorescence. Human-rhesus chimeric liver can be established using adult bone marrow mesenchymal stem cells, which can generate functional liver cells in the liver of infant rhesus.

Objective:To evaluate the clinical application value of gastrointestinal contrast-enhanced ultrasound combined gas injection method in verifying the location of nasointestinal tube in critically ill patients.Methods:Data of 60 critically ill patients who had the indications of indwelling nasointestinal tube were collected from September 1,2015 to September 1,2016 in the Intensive Care Unit of Zhejiang Provincial People(s) Hospital.The position of nasointestinal tube in patients who underwent bedside blind insertion would be confirmed routinely through gas injection auscultation method.After tube was inserted,its route was scanned by ultrasound with gas perfusion assistance.Afterwards,rapid gas perfusion was used until suspicious tube end position was determined.Furthermore,oral ultrasound contrast agent was injected into the tube if instantaneous strong echo of gas was observed in localized lumen,and contrast agent filling meant the placement being successful.Two methods of position confirmation of nasointestinal tube in critically ill patients included gastrointestinal contrast enhanced ultrasound combined gas injection and gas injection auscultation only,and the effect of the two methods was compared and confirmed by chest and abdominal X ray examinations to verify the location of nasointestinal tube below pylorus.Results:A total of 60 patients were included in this study,58 patients(96.7％)in gastrointestinal contrast enhanced ultrasound combined gas injection group were successfully positioned.Among them,the placements of tube in 56 cases were below pylorus,while 2 cases were above pylorus.The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of location of gastrointestinal contrast enhanced ultrasound combined gas injection method were 96.6％,100％,100％,50％,96.7％ and of gas injection auscultation method were 74.1％,50％,97.7％,6.3％ and 73.3％.The differences of the sensitivity,specificity,negative predictive value and accuracy between the two methods were statistically significant (P ＜ 0.05).Conclusion:Gastrointestinal contrastenhanced ultrasound combined gas injection method is a safe,simple and convenient method with high sen-sitivity,specificity,negative predictive value and accuracy in confirming the location of the nasointestinal tube.

Purpose To investigate the effects of chemotactic factor CCR4 on the abi1ity of pro1iferation,ce11 cyc1e,invasion,and mi-gration of human ga11b1adder cancer ce11. Methods Western b1ot was used to detect the expression 1eve1 of CCR4 in ga11b1adder carci-noma ce11s. Ga11b1adder carcinoma ce11s was infected by means of s1ow virus,the CCR4 gene si1encing was conducted using siRNA-CCR4 interference techno1ogy. Ga11b1adder carcinoma ce11s GBC-SD were divided into three groups( GBC-SD,GBC-SD/CCR4-RNAi and GBC-SD/contro1). CCL17,a 1igand of CCR4,was used to act on these three groups of ce11s. CCK8 method was used to detect the ce11 pro1iferation abi1ity of three groups. F1ow cytometry was used to test ce11 cyc1e. Tanswe11 assay was app1ied to detect ce11 migration and invasion abi1ity. Western b1ot was performed to detect the expression of its corresponding 1igands CCL17 and CCL22 proteins. Re-sults CCR4 gene si1ence did not inf1uence ce11 cyc1e and pro1iferation of ga11b1adder ce11 GBC-SD,but can significant1y inhibit GBC-SD ce11 invasion and movement abi1ity,CCR4 gene si1ence had no inf1uence on the expression of CCL17 and CCL22 gene in tumor ce11s. Conclusion Ga11b1adder carcinoma ce11s GBC-SD express chemokine receptor CCR4,chemokine receptor CCR4 can promote the invasion and metastasis of GBC-SD ce11s.

Objective To evaluate the effect of BinaxNOW rapid diagnostic devices for malaria in the field. Methods The symptomatic patients were detected by using the BinaxNOW devices and the results were compared with the microscopic examination of Giemsa-stained blood smears(as a golden standard). The sensitivity and specificity were calculated. The double-blind method was used in this study. Results In 443 symptomatic patients,151 cases were positive for malaria. The sensitivity was 98.69% and the specificity was 100%,and the coincidence rate of BinaxNOW with the microscopic examination was 99.55%. The sensitivity and specificity of Plasmodium falciparum were 100% and 99.66%,respectively,and the coincidence rate was 99.76%. The sensitivity and specificity of non-falciparum Plasmodium were 90.91% and 100%,respectively,and the coincidence rate was 99.36%. Conclusion The BinaxNOW rapid diagnostic devices for malaria are sensitive,specific,and stable for malaria diagnosis in the field.

Objective To investigate the value of urinary liver-type fatty acid-binding protein (L-FABP),neutrophil gelatinase-associated lipocalin (NGAL) and their combination in predicting the development and the severity of acute kidney injury (AKI) following cardiac surgery in adults. Methods Scr,urinary L-FABP and NGAL corrected by urine creatinine at preoperation,0 h and 2 h postoperative time points were examined.The differences of above indexes between AKI and non-AKI groups were compared.Receiver operating characteristic (ROC)curves and area under curves (AUC) were used to evaluate the diagnostic value of urinary L-FABP,NGAL and their combination for AKI. Results The cohort consisted of 109 patients,26(23.9％) developed AKI,and AKIN stage Ⅰ, Ⅱ and Ⅲ was 46.2％,34.6％ and 19.2％ respectively.Levels of urinary L-FABP and NGAL were significantly higher in AKI patients at 0 h and 2 h postoperatively.AUC to predict AKI or AKI stage Ⅱ-Ⅲ was 0.81 to 0.87 using either of the biomarkers.The performance of combining two biomarkers was better with AUC of 0.911 to 0.927. Conclusions Urinary L-FABP and NGAL increase at the early stage after cardiac surgery.Combination of these two biomarkers enhances the accuracy of the early diagnosis of postoperative AKI after cardiac surgery before a rise of Scr.

Objective:To investigate the role of LncRNA-TUG1 in the injury of intestinal epithelial cells induced by lipopolysaccharide (LPS).Methods:LPS was used to treat HIEC-6 human intestinal epithelial cells for 24 h to construct a sepsis injury model. Whole transcriptome RNA sequencing was used to analyze the expression changes of mRNA, microRNA and lncRNA in HIEC-6 cells after LPS treatment. Real-time fluorescence quantitative (qRT-PCR) and Western blot was performed to detect the expression changes of lncRNA-TUG1, microRNA-132-3p (miR-132-3p), SIRT1 mRNA and SIRT1 protein in HIEC-6 cells after LPS treatment. The expression levels of LncRNA-TUG1, miR-132-3p and SIRT1 were artificially changed by in vitro transfection. qRT-PCR and Western blot were used to confirm the regulatory effect of lncRNA-TUG1 on microRNA-132-3p and SIRT1. CCK-8 and flow cytometry were used to analyze the effects of LncRNA-TUG1, miR-132-3p and SIRT1 on the proliferation and apoptosis of HIEC-6 cells. The dual luciferase report analysis was used to verify the targeting relationship between LncRNA-TUG1, miR-132-3p and SIRT1. Statistical analysis was performed using SPSS 17.0, and differences between the two groups were compared using independent sample t test. Results:RNA sequencing results showed that the expressions of lncRNA-TUG1 and SIRT1 were decreased in HIEC-6 cells after LPS treatment ( t=3.26, P<0.05 and t=2.55, P<0.05), but the expression of miR-132-3p was increased ( t=4.12, P<0.05). In vitro cell experiments, the expression of lncRNA-TUG1 and SIRT1 were decreased in HIEC-6 cells treated with LPS ( t=5.69, P<0.05 and t=5.712, P<0.05), while the expression of miR-132-3p was increased ( t=3.88, P<0.05). Overexpression of lncRNA-TUG1 increased the proliferation rate ( t=6.55, P<0.05) and decreased the apoptosis rate ( t=3.94, P<0.05) of LPS-treated cells. Upregulation of lncRNA-TUG1 decreased the expression of miR-132-3p ( t=4.66, P<0.05), and increased the mRNA and protein levels of SIRT1 ( t=3.91, P<0.05). Transfection of miR-132-3P mimic could inhibit the mRNA ( t=4.08, P<0.05) and protein levels of SIRT1. In LPS-treated cells, the cells co-transfected with miR-132-3pmimic and siRNA-SIRT1 had a lower proliferation rate ( t=4.55, P<0.05 and t=5.67, P<0.05) and a higher apoptosis rate ( t=3.90, P<0.05 and t=4.22, P<0.05) than those transfected with only pcDNA3.1-lncRNA-TUG. Conclusions:lncRNA-TUG1 may act as a ceRNA to regulate miR-132-3p/SIRT1, therefore alleviating HIEC-6 cell injury caused by LPS. Intervention of lncRNA-TUG1/miR-132-3p/SIRT1 regulatory pathway may become a potential strategy to prevent sepsis-induced intestinal mucosal damage.

Objective To explore the relationship of phospholipase A2 receptor (PLA2R) expression in renal tissue with clinical characteristics,prognosis of idiopathic membranous nephropathy (IMN) patients.Methods 134 patients diagnosed as nephropathy proven by biopsy was selected as subjects of this research,including 98 patients with IMN patients,10 patients with secondary membranous nephropathy and 26 patients with other renal glomerular diseases.The expression of PLA2R antigen in renal tissue was detected by immuno-fluorescence chemistry staining.Results The positive rate of renal PLA2R expression in IMN patients was higher than that in SMN patients (91.84％ vs 40.00％,P＜ 0.01),whereas there is no expression in other glomerular diseases.The PLA2R negative group were mainly stage Ⅰ membranous nephropathy,and positive group was mainly in stage Ⅱ.The distribution of pathological stage between the two groups was statistically significant (P ＜0.01).Compared with the positive group,the negative group was manifested with higher eGFR[(115.91± 23.32) ml· min-1 · (1.73 m2)-1 vs (94.06±27.38) ml· min-1 · (1.73 m2)-1,P=0.031],associated with the higher 12-month complete remission rate (87.50％ vs 44.07％,P=0.021).Conclusions The expression of PLA2R antigen in renal tissue plays an important role in the diagnosis,disease evaluation and prognosis of IMN.The negative PLA2R in kidney tissue of IMN may indicate a good clinical prognosis.