OBJECTIVES: To investigate the role of sphingosine-1-phosphate receptor 5 (S1PR5) in modulating barrier function of mouse brain microvascular endothelial cells with oxygen-glucose deprivation and reoxygenation (OGD/R). METHODS: Mouse brain microvascular endothelial cells (bEnd.3) were exposed to OGD/R to induce barrier dysfunction following treatment with S1PR5-specific agonist A971432 or lentivirus-mediated transfection with a S1PR5-specific siRNA, a S1PR5-overexpressing plasmid, or their respective negative control sequences. The changes in viability and endothelial barrier permeability of the treated cells were evaluated with CCK-8 assay and FITC-dextran permeability assay; the levels of intracellular reactive oxygen species (ROS) and localization and expression levels of the proteins related with barrier function and oxidative stress were detected using immunofluorescence staining, DCFH-DA probe and Western blotting. RESULTS: S1PR5 activation obviously enhanced viability of bEnd.3 cells exposed to OGD/R (P<0.0001). Both activation and overexpression of S1PR5 reduced FITC-dextran leakage, while S1PR5 knockdown significantly increased FITC-dextran leakage in the exposed bEnd.3 cells. Activation and overexpression of S1PR5 both increased the cellular expressions of the barrier proteins ZO-1 and occludin, while S1PR5 knockdown produced the opposite effect. In cells exposed to OGD/R, ROS production was significantly reduced by S1PR5 activation and overexpression but increased following S1PR5 knockdown. Overexpression of S1PR5 obviously increased the expressions of the antioxidant proteins Nrf2, HO-1 and SOD2 in the exposed cells. CONCLUSIONS S1PR5 activation and overexpression significantly improve cell viability and reduce permeability of a mouse brain microvascular endothelial cell model of OGD/R, the mechanism of which may involve the reduction in ROS production and upregulation of the antioxidant proteins.
Animals ; Mice ; Oxidative Stress ; Endothelial Cells/cytology* ; Brain/blood supply* ; Reactive Oxygen Species/metabolism* ; Receptors, Lysosphingolipid/metabolism* ; Sphingosine-1-Phosphate Receptors ; Blood-Brain Barrier/metabolism* ; Glucose ; Cell Line ; Oxygen/metabolism* ; NF-E2-Related Factor 2/metabolism*

Objective:To investigate the biological behavior spectrum of platelet-derived growth factor alpha receptor (PDGFRA)-mutant gastrointestinal stromal tumor (GIST), and to compare the clinical values of the Zhongshan method of benign and malignant evaluation with the modified National Institutes of Health (NIH) risk stratification.Methods:A total of 119 cases of GIST with PDGFRA mutation who underwent surgical resection at Zhongshan Hospital, Fudan University from 2009 to 2020 were collected. The clinicopathological data, follow-up records, and subsequent treatment were reviewed and analyzed statistically.Results:There were 79 males and 40 females. The patients ranged in age from 25 to 80 years, with a median age of 60 years. Among them, 115 patients were followed up for 1-154 months, and 13 patients progressed to disease. The 5-year disease-free survival (DFS) and overall survival (OS) were 90.1% and 94.1%, respectively. According to the modified NIH risk stratification, 8 cases, 32 cases, 38 cases, and 35 cases were very-low risk, low risk, intermediate risk, and high risk, and 5-year DFS were 100.0%, 95.6%, 94.3%, and 80.5%, respectively. There was no significant difference in prognosis among the non-high risk groups, only the difference between high risk and non-high risk groups was significant ( P=0.029). However, the 5-year OS was 100.0%, 100.0%, 95.0% and 89.0%, and there was no difference ( P=0.221). According to the benign and malignant evaluation Zhongshan method, 43 cases were non-malignant (37.4%), 56 cases were low-grade malignant (48.7%), 9 cases were moderately malignant (7.8%), and 7 cases were highly malignant (6.1%). The 5-year DFS were 100.0%, 91.7%, 77.8%, 38.1%, and the difference was significant ( P<0.001). The 5-year OS were 100.0%, 97.5%, 77.8%, 66.7%, the difference was significant ( P<0.001). Conclusions:GIST with PDGFRA gene mutation shows a broad range of biological behavior, ranging from benign to highly malignant. According to the Zhongshan method, non-malignant and low-grade malignant tumors are common, the prognosis after surgery is good, while the fewer medium-high malignant tumors showed poor prognosis after surgical resection. The overall biological behavior of this type of GIST is relatively inert, which is due to the low proportion of medium-high malignant GIST. The modified NIH risk stratification may not be effective in risk stratification for PDGFRA mutant GIST.

【Objective】 In this study， reactive oxygen species （ROS） scavenger N-acetyl-L-cysteine （NAC） was used to explore the inhibitory effect and mechanism of artesunate （ART） on pancreatic carcinoma （PC） cells. 【Methods】 Different concentrations of ART interfered with 3 PC cell lines CFPAC-1， Capan-2 and BxPC3. Cell viability was measured by CCK8； cell migration ability was measured by Transwell method， and the expressions of migration-related proteins E-cadherin， N-cadherin and Vimentin were measured by Western blotting. ROS probe DCFH-DA was used to measure intracellular ROS； LC3 cell immunofluorescence （IF） was used to detect the formation of intracellular autophagosomes. After adding NAC or autophagy inhibitor 3-MA， the cell viability was tested again by CCK8， and the expressions of p-AMPK/ AMPK， p-mTOR/mTOR， p62 and LC3Ⅱ/Ⅰ were detected by Western blotting. 【Results】 ART inhibited the growth of CFPAC-1 and Capan-2 in a time- and dose-dependent manner. After treatment of CFPAC-1 and Capan-2 cells with 200 μmol/L of ART for 48 h， the expression of E-cadherin was upregulated， while N-cadherin and Vimentin were downregulated， and the cell migration ability was significantly reduced. ART significantly upregulated intracellular ROS level and promoted the formation of autophagosomes. NAC could reduce the inhibitory effect of ART on CFPAC-1 and Capan-2 cells， upregulate p-AMPK/AMPK， P62 and LC3Ⅱ/Ⅰ， downregulate the expression of p-mTOR/mTOR， and intensify autophagy. 3-MA could not reverse the inhibitory effect of ART on PC cells. 【Conclusion】 ART is dependent on ROS， but not on autophagy， in exerting an anti-pancreatic carcinoma effect. NAC attenuates the inhibitory effect of ART on PC cells by activating protective autophagy through AMPK/mTOR signaling pathway.

Objective To explore the efficacy and safety of mechanical thrombectomy with Solitaire device in patients with acute basilar artery occlusion (ABAO).Methods The clinical data of 5 patients with ABAO,underwent mechanical thrombectomy with Solitaire stent in our hospital from May 2016 to November 2018,were analyzed retrospectively.The clinical treatment efficacy and safety were recorded.Results All these 5 patients accepted endovascular mechanical thrombectomy successfully;successful recanalization (modified Thrombolysis in Cerebral Infarction [mTICI] grading 2b or 3) was achieved in 4 patients;the embolus was detached to the distal arteriole in one patient.There was no symptomatic intracranial hemorrhage;one patient accepted decompressive craniectomy resulting from massive infarction of cerebellum.Follow up results 90 d after treatment indicated that 4 patients enjoyed good prognosis (modified Rankin Scale scores:0-3).Conclusion Mechanical thrombectomy with Solitaire device is suggested to be safe and efficient in ABAO patients complying with appropriate operative indications.

ObjectiveTo explore the nursing students' experience of participation in group resuscitation workshop. MethodsFrom July to September of 2018,12 nursing interns in the Emergency Department of a Class Ⅲ Grade A hospital in Zhengzhou were recruited to participate in the study by purposive sampling and semi-structured interviews were conducted. The data obtained were analyzed by Colaizzi method. ResultsNursing students' experience of participation in the team recovery workshop can be summarized into three themes:teamwork awareness, independent learning ability and confidence,and stress and response. ConclusionsThe workshop promotes nursing students' sense of teamwork and stimulates their subjective initiative and ability to learn. It is suggested that nursing clinical education workers should attach importance to the various pressures faced by students and provide support from various angles.

Objective: To observe the analgesic and anti-inflammatory effect of mandelic acid. Methods: Fifty Kunming mice were randomly divided into 5 groups:the blank control group (0. 1 ml/10 g), mandelic acid high (300 mg·kg-1), medium (200 mg ·kg-1 ) and low (140 mg·kg-1 ) dose groups, and the positive control ( aspirin) group, ig, qd. The analgesic effect of mandelic acid was observed by writhing test and hot plate method in mice. The ear swelling model caused by dimethyl benzene in mice was a-dopted to observe the analgesic effect. Results:Mandelic acid in each dose group could make the number of writhing in mice signifi-cantly reduced and pain threshold extended, and when compared with the blank control group, the difference was statistically significant (P<0. 01). The writhing times of mice mandelic acid high dose group was fewer than that of the positive control group, and there was no statistically significant between the groups (P>0. 05). In low and medium dose group, the writhing times of mice were more than those of the positive control group, and there was a significant difference between the low dose group and the positive control group( P<0. 05). The pain threshold of the mice in each mandelic acid dose group was higher than that of the positive control group, the pain threshold increased significantly in the high dose group before and after the administration, and the difference was statistically signifi-cant when compared with the positive control group (P<0. 05). The effect of mandelic acid on the ear swelling of mice was not signifi-cant, and when compared with the blank control group, the difference was not significant (P>0. 05). Conclusion:Mandelic acid has significant analgesic effect, while anti-inflammatory effect is not obvious.

Objective To discuss the reason of skin flap necrosis caused by vascular crisis of reverse island flap of forearm posterior interosseous artery.Methods Eight-six patients who were underwent reverse island flap of forearm interosseous posterior artery for deep tissues and skin defect on the back of hand between March,2002 and April,2014 were analyzed in this study.Eleven patients had occurred skin flap necrosis,include 5 cases had completely flap necrosis caused by circulation crisis,and 6 cases had partial necrosis at the distal of the flap.Among the necrosis cases,5 cases were injured by the machine injury,4 cases by the heavy crush and 2 cases by the traffic accident.The cause of circulation crisis was analyzed.Results In the series,75 skin flaps survived completely and 11 cases had occurred necrosis,included completely necrosis with 5 cases.The reasons of flap crisis were as follows:for the completely necrosis,2 cases with variation of perforating branch of posterior interosseous artery,1 case with absence of posterior interosseous artery,1 case with vessel pedicel entrapment in subcutaneous tunnel,and 1 case with misconduct venous congestion caused by the reverse perfusion of superficial vein.The reason of circulation crisis of completely necrosis were as follows:2 cases with artery crisis and 3 of them with distortion of entrapment at pedicel and vein crisis.One case was cured through debridement,change of medical prescription and skin grafting;and 4 cases were cured with other flap repair technique.For the partial necrosis,2 cases with variation of perforating branch of posterior interosseous artery,1 case with excessively narrow entrapment at pedicel in subcutaneous tunnel,1 case with folding vessel pedicel entrapment of skin at the back of wrist,1 case with misconduct of superficial vein trunk and 1 case with intraoperative side-injury.The symptoms of circulation crisis of completely necrosis were as follows:2 cases with artery crisis and 4 of them with distortion of entrapment at pedicel and vein crisis.Four cases were cured through debridement and skin grafting,1 case was cured by the vacuum-sealing drainage (VSD) and 1 case with skin flap repair at pedicle of abdomen.Conclusion The anatomic variation of perforator vessel of reverse island flap of forearm posterior interosseous artery;narrow entrapment at pedicel in subcutaneous tunnel and distortion of entrapment at pedicel;venous congestion caused by the reverse perfusion of superficial vein;intraoperative side-injury of the pedicel of the flap;excessively folding vessel pedicel entrapment of skin at the back of wrist after surgery will cause the circulation crisis of reverse island flap of forearm posterior interosseous artery and induce the necrosis of the skin flap.

BACKGROUND:There is a close relationship between epilepsy and apoptosis. The appearance of epilepsy can lead to the loss of neurons in the hippocampus, triggering a series of programmed cel death. OBJECTIVE:To investigate the effect of bone marrow stromal stem cel transplantation on apoptosis in epilepsy. METHODS:After modeled to be of epilepsy 45, Sprague-Dawley model rats were randomly divided into three groups, fol owed by given no intervention (moldel group), normal saline (normal saline group) or bone marrow stromal stem cel transplantation (transplantation group). At 1, 2 and 4 weeks after modeling, the number of Bax-positive cel s, Bcl-2-positive cel s and Bax/Bcl-2 were detected by immunohistochemistry. RESULTS AND CONCLUSION:The number of Bax-positive cel s, Bcl-2-positive cel s and Bax/Bcl-2 presented no obvious changes in the normal saline group at different time points. However, the number of Bax-positive cel s and Bax/Bcl-2 in the transplantation group was significantly decreased, while the number of Bcl-2-positive cel s significantly increased compared with the other two groups at 1, 2 and 4 weeks after modeling (P<0.05). Moreover, the above indicators varied significantly in the transplantation group at different time points after modeling (P<0.05). These results show that bone marrow stromal stem cel transplantation can affect the apoptosis and effectively reduce the apoptosis in rats with epilepsy by up-regulating the number of Bax-positive cel s and down-regulating the number of Bcl-2-positive cel s.

Objective To investigate the effect of terlipressin on hepatic and renal function in cirrhotic patients undergoing hepatectomy.Methods Aanlyze the clinical data of 57 patients following irregular hepatectomy for hepatocellular carcinoma with cirrhosis,according to whether use terlipressin or not after operation,which were divided into terlipressin group (A group,n =27) and control group (B group,n =30).Liver function parameters (ALT,AST,TB),ascites,urine volume and renal function parameters (Cr,BUN) preoperatively and on postoperative day(POD) 1,3,5 and 7 were compared between the two gruops.Results Compared with those of POD 1,the levels of ALT,AST and ascites on POD 3,5,7 were significantly lower in two groups (P ＜ 0.05),urine volume was significantly increased (P ＜ 0.05),Cr of POD 7 was significantly lower (P ＜0.05),but it is more remarkable in group A than group B.The levels of ALT in terlipressin group on POD 5,7 were (144.9 ±76.3) U/L,(100.5 ±61.5) U/L,which were lower than those of (267.2±91.2) U/L,(199.3 ±70.5) U/L in control group.On POD 3,5,7,the levels of AST,BUN,Cr and peritoneal fluid in terlipressin group,which were respectively(211.1 ±99.8) U/L,(80.4 ±54.6) U/L,(50.6 ±46.5) U/L,(6.6 ± 1.9) mmol/L,(6.5 ± 1.7) mmol/L,(6.3 ± 2.1) mmol/L,(74.3 ± 10.9) μmol/L,(71.5 ± 8.9) μmol/L,(58.7 ±4.1) μmol/L,(247.6±60.3) ml,(58.8±54.3) ml,(40.2±31.8) ml,were significantly lower than those in control group which were (298.7 ±131.2) U/L,(201.1 ±93.4) U/L,(114.7 ±70.3) U/L,(7.3 ± 1.9) mmol/L,(7.2±1.8) mmol/L,(7.1±1.7) mmol/L,(79.5±15.1) μmol/L,(76.9±16.2) μmol/L,(69.4±11.4) μmol/L,(275.2±88.1) ml,(191.7±71.6) ml,(93.2±50.2) ml.while urine volume of (2232.3±409.8) ml,(2270.5 ±395.8) ml,(2179.0±301.4) ml was much more than that of (1921 ± 510.4) ml,(2019.1 ±411.2) ml,(1978.7±323.7) ml in the control group,the differences in the two groups were statistically significant (P ＜ 0.05).There were 11 (36.7％) patients with hepatic and renal dysfunction and hepatorenal syndrome after operation in group B,while only 2 (7.4％) patients in group A.Conclusions The use of terlipressin after partial liver resection has a protective effect on hepatic and renal function in patients with cirrhosis,and can reduce postoperative ascites and prevent hepatorenal syndrome.

Objective To explore the effect of donepezil combined with memantine in ameliorating the cogni-tive function and psychological behavior of patients with Alzheimers'disease(AD).Methods 214 AD patients were randomly divided into the control groupand observation group.The control group were treated with memantine,and the observation group were treated donepezil combined with memantine.Before and after treatment,MMSE and WHO -UCLA AVLT,BEHAVE -AD were utilized to assess the cognitive function,psychological behavior,respectively.The adverse effects were observed using TESS and laboratory examination.Results The scores of MMSE and its subitem, WHO -UCLA AVLT(Immediate memory,short -term memory and long -term memory)after treatment significantly increased as compared with those before treatment in the control group and the observation group(P <0.01 or P <0.05),while the cases of psychological behavior,the score of BEHAVE -AD and its subitem decreased(P <0.01 or P <0.05).The observation group showed statistically significant difference in the increasion of the scores of MMSE and its subitem,WHO -UCLA AVLT,and the decreasion of the cases of psychological behavior,the score of BE-HAVE -AD and its subitem comparison with the control group(t =-1.742,-1.431,-1.626,-2.173,-1.448,-1.026,-1.016,-1.279,-1.971,χ2 =12.295,7.609,8.917,t =0.778,1.139,2.004,1.153,0.822,1.944, P <0.01 or P <0.05).The total effective rate of the observation group was 90.65%,and 71.96% of the control group,there was a significant difference between the two groups(χ2 =12.299,P <0.01).There was no obvious side effects during the treatment in both groups.Conclusion Donepezil combined with memantine can significantly improve the cognitive function and psychological behavior of AD patients,and there are fewer adverse events.