Objective To establish a chronic rejection (CR) model of mouse heart transplantation and analyze its characteristics. Methods Allogeneic BALB/c and C57BL/6 mice were used as donor and recipient for heart transplantation, and intraperitoneal injection of cytotoxic T lymphocyte-associated antigen 4-immunoglobulin (CTLA4-Ig) was given 1 and 2 days after surgery. Graft survival time, donor specific antibody (DSA) level, graft pathology and inflammatory cell infiltration were observed. Results In allogeneic transplantation model, graft survival time was prolonged after CTLA4-Ig treatment [(28.2±4.1) d vs. (7.0±0.7) d, P < 0.01]. The level of serum DSA-IgG increased at 2, 3 and 4 weeks after surgery, while the level of DSA-IgM remained unchanged. Myocardial cell injury, inflammatory cell infiltration, interstitial fibrosis and C4d deposition in capillaries were aggravated 3 weeks after operation and worsened 4 weeks after operation. The infiltrated immune cells were mainly macrophages, T cells and plasma cells. Conclusions Mouse allogeneic heart transplantation combined with CTLA4-Ig successfully establishes a CR model, which provides a basis for subsequent studies on the pathogenesis and intervention of CR.

According to the "Regulations on clinical application management of medical technologies", physicians intending to carry out restricted technologies must undergo standardized training and pass assessments in accordance with the clinical application management standards for the respective technology. As ventricular assist technology is classified as a nationally restricted technology, standardized training is one of the essential conditions for its application. This paper primarily explores the standardized training for the clinical application of ventricular assist technology in Shanghai, in light of its background, clinical application, and current training status. It proposes the training requirements for ventricular assist technology, animal training assessment standards, and clinical practice assessment standards in Shanghai, aiming to promote the standardized development and high-quality advancement of ventricular assist technology in Shanghai.

Objective Utilizing a specially engineered miR-144-GFP-H1 human embryonic stem cell(hESC)reporter line,this study leverages GFP fluorescence as an indicator of miR-144 expression to gauge the progression of erythropoiesis.The investigation is aimed at elucidating the potential roles of lncRNAs within the erythropoietic framework and conducting an initial assessment of their functional impact.Methods The miR-144/451-GFP-H1 cell line(hereafter referred to as 144-H1)was utilized for in vitro erythrocyte induction culture.The subpopulations of cells entering the erythropoiesis stage were characterized by the surface molecules CD71 and GPA.The GFP reporter gene of miR-144 served as a critical determi-nant to distinguish between GFP-positive cells(with a high propensity for erythropoiesis)and GFP-negative cells(with a low propensity for erythropoiesis).Transcriptome sequencing was performed on both groups to identify differentially ex-pressed long non-coding RNAs(lncRNAs).LncRNA entries with potential for validation were selected for preliminary func-tional verification.The CRISPR/Cas9 gene editing technique was employed to design functional interference strategies for the targeted lncRNAs,obtaining 144-H1 cell lines with knocked-out function of the specific lncRNAs.These knockout cell lines,along with non-knockout 144-H1 cell lines,were used for parallel erythrocyte induction culture to identify differential nodes.This approach preliminarily verified their impact on erythropoiesis in an in vitro development model.Results 1)The constructed 144-H1 cell line was capable of expressing GFP fluorescence upon entering the stage of in vitro erythrocyte in-duction,indicating the activation of miR-144/451.2)Within the CD71,GPA double-positive group,significant differences in lncRNA expression were observed between the GFP-positive and GFP-negative subpopulations.3)Gene editing strategies involving the deletion of sequence segments capable of effectively interfering with the function of multiple lncRNA entries were designed and verified for successful editing.In the knockout cell lines,parts of the lncRNA sequences were directly de-leted.4)In parallel validation experiments of erythrocyte induction culture,cell lines with LINC01569 knocked out exhibited significant differences in flow cytometric subpopulations and cell proliferation capabilities compared to the non-knockout cell lines:①The knockout cell lines showed sustained high expression of GFP fluorescence.②The proportion of the CD71-GPA double-positive group in the knockout cell lines continuously decreased during erythrocyte maturation.③No significant ex-pression of hemoglobin was observed in the knockout cell lines,lacking the characteristic red color.④The cell proliferation capability of the knockout cell lines was significantly lower than that of the non-knockout cell lines(P＜0.05).Conclusion The successful employment of the 144-H1 cell line facilitated an exploration into the potential functions of lncRNAs in e-rythropoiesis.This enables the design of more refined in vitro developmental experiments to enhance the precision in captu-ring lncRNA functions.Among the differentially expressed lncRNA entries,LINC01569 was preliminarily validated to play a regulatory role in erythropoiesis.The functional absence of LINC01569 severely impacts the normal differentiation and prolif-eration of erythrocytes.The specific regulatory mechanism of LINC01569 in erythropoiesis warrants further investigation and research.

Pheochromocytomas and paragangliomas(PPGLs)cause symptoms by altering the circulation levels of catecholamines and peptide hormones.Currently,the diagnosis of PPGLs relies on diagnostic imaging and the detection of catecholamines.In this study,we used ultra-performance liquid chromatography(UPLC)/quadrupole time-of-flight mass spectrometry(Q-TOF MS)analysis to identify and measure the perioperative differential metabolites in the plasma of adrenal pheochromocytoma patients.We identified differentially expressed genes by comparing the transcriptomic data of pheochromocytoma with the normal adrenal medulla.Through conducting two steps of metabolomics analysis,we identified 111 differential metabolites between the healthy group and the patient group,among which 53 metabolites were validated.By integrating the information of differential metabolites and differentially expressed genes,we inferred that the cysteine-methionine,pyrimidine,and tyrosine metabolism pathways were the three main metabolic pathways altered by the neoplasm.The analysis of transcription levels revealed that the tyrosine and cysteine-methionine metabolism pathways were downregulated in pheochromocytoma,whereas the pyrimidine pathway showed no significant difference.Finally,we developed an optimized diagnostic model of two metabolites,L-dihydroorotic acid and vanylglycol.Our results for these metabolites suggest that they may serve as potential clinical biomarkers and can be used to supplement and improve the diagnosis of pheochromocytoma.

Objective To evaluate the effect of hydrogen sulfide on myocardial exogenous apoptotic pathway in a rat model of hemorrhagic shock and resuscitation.Methods Sixty clean-grade healthy male Sprague-Dawley rats,aged 8 weeks,weighing 250-300 g,were divided into 4 groups (n =15 each) using a random number table method:sham operation group (group S),sham operation plus sodium sulphid (NaHS) group (group S+NaHS),hemorrhagic shock group (HS group),and hemorrhagic shock plus sodium sulphid group (group HS+NaHS).Rats only underwent arterial and intravenous puncture in group S.Hemorrhagic shock was induced by withdrawing blood from the femoral artery until mean arterial pressure (MAP) was reduced to 35-40 mmHg within 10 min and maintained for 1.5 h.NaHS 28 μmol/kg was intraperitoneally injected at 10 min before resuscitation in group HS+NaHS.The equal volume of NaHS was administered at the same time in group S+NaHS.Immediately before blood letting and at 0,1.5,2,3,4 and 6 h after blood letting (T1-5),MAP was recorded and blood samples were collected from the femoral vein for determination of serum creatine kinase (CK) and lactate dehydrogenase (LDH) concentrations by chemical colorimetry.Rats were then sacrificed and hearts were removed for examination of the pathological changes of myocardial tissues (with a light microscope) and for determination of cell apoptosis (by TUNEL),expression of caspase-3 and caspase-8 (by Western blot) and expression of Fas and FasL (by immunohistochemistry).Apoptosis index was calculated.Results Compared with group S,MAP was significantly decreased at T1-5,the serum CK and LDH concentrations at T1-5 and apoptosis index at T5 were increased,and the expression of Fas,FasL,caspase-3 and caspase-8 was up-regulated in group HS (P＜ 0.05).Compared with group HS,MAP was significantly increased at T1-3,the serum CK and LDH concentrations at T3-5 and apoptosis index at T5 were decreased,and the expression of Fas,FasL,caspase-3 and caspase-8 was down-regulated in group HS+NaHS (P＜0.05).The pathological changes of myocardial tissues were significantly attenuated in group HS+NaHS when compared with group HS.Concclusion The mechanism by which hydrogen sulfide attenuates myocardial injury induced by hemorrhagic shock and resuscitation is associated with inhibiting the exogenous apoptotic pathway in rats.

Objective To evaluate the role of nitric oxide in sevoflurane postconditioning-induced mitigation of autophagy and cell apoptosis during ischemia/reperfusion (I/R) in isolated rat hearts.Methods The hearts of male Sprague-Dawley rats,aged 2-3 months,weighing 250-300 g,were excised and retrogradely perfused in a Langendorff apparatus.One hundred and eight isolated rat hearts,which were successfully perfused in a Langendorff apparatus,were equally and randomly divided into 6 groups:control group (C group),sevoflurane group (S group),I/R group,sevoflurane postconditioning group (SSP group),sevoflurane postconditioning + L-NAME (non-selective nitric oxide synthase (NOS) inhibitor group (SSP + L group),and L-NAME group (L group).The hearts were perfused with K-H solution for 150 min in C group.The hearts were continuously perfused for 180 min and perfused with K-H solution containing 3％ sevoflurane for 15 min starting from 60 min of perfusion in S group.After being perfused with K-H solution for 30 min,the hearts were subjected to occlusion for 30 min followed by reperfusion for 120 min in the other groups except C and S groups.After onset of reperfusion,the hearts were perfused with K-H solution containing 3％ sevoflurane for 15 min in SSP group,the hearts were perfused with K-H solution containing 3％ sevoflurane and L-NAME 100 μmol/L for 15 and 60 min,respectively,in SSP + L group,and the hearts were perfused with K-H solution containing L-NAME 100μmol/L for 60 min in L group.Inn ediately before ischemia,and at 30,60,90 and 120 min of reperfusion,each parameter of cardiac function was recorded.At the end of reperfusion,myocardial specimens were obtained at the end of reperfusion for measurement of the infarct size,NOS activity,NO content,and expression of Bcl-2,Beclin 1 and caspase-3,for observation of formation of autophagosomes,and for examination of the pathological changes.Results Compared with C group,LVSP,+ dp/dtmax,-dp/dtmax,NOS activity and NO content were significantly decreased,and LVEDP was increased in I/R and SSP groups.Compared with I/R group,LVSP,+ dp/dtmax,-dp/dtmax,NOS activity and NO content were significantly increased,LVEDP was decreased,Bcl-2 expression was down-regulated,and the expression of Beclin 1 and caspase-3 was up-regulated in SSP group,and no significant changes were found in each index in SSP+ L and L groups.Compared with SSP group,LVSP,+ dp/dtmax,-dp/dtmax,NOS activity and NO content were significantly decreased,LVEDP was increased,Bcl-2 expression was down-regulated,and the expression of Beclin 1 and caspase-3 was up-regulated in SSP + L group.Conclusion The mechanism by which sevoflurane postconditioning reduces I/R injury may be related to promoted NO product and inhibited autophagy and cell apoptosis in isolated rat hearts.

Objective:To explore how coping style and social support influence the quality of life in patients with impaired glucose tolerance, which act respectively as the internal and external mediating ways. Methods:A total of 283 patients with impaired glucose tolerance from 6 Three-A hospitals in China were surveyed with self-rating anxiety scale, self-rating depression scale, trait coping style questionnaire, social support scale, and WHOQOL-BREF.
Results:Biographic data failed to predict the quality of life in patients with impaired glucose tolerance, while anxiety, depression, social support and coping style significantly influenced their quality of life.
Conclusion:The fact that emotional disorder, social support and coping style influence the quality of life in patients with type 2 diabetes also exists in patients with impaired glucose tolerance.

ObjectiveTo explore the expression of Glut1, HIF-1α and Ki-67 in hepatocellular carcinoma (HCC) and their relationship between their expression and clinicopathological features.MethodsImmunohistochemical study (EnVision method) for Glut1, HIF-1a and Ki-67 were performed on tissue microarray which consisted of 171 cases of HCC, 55 cases of adjacent non-neoplastic liver tissues, and 22cases of normal liver tissues.ResultsThe expression rate of Glut1 ,HIF-1α and ki-67 in 171 cases of HCC was 15.2％, 19.9％ and 66.1％, respectively, which was much higher than that in the adjacent non-neoplastic liver tissues (1.8％ ,1.8％and 5.5％) and normal liver tissues(all negative).The expressions of Glut1 and HIF-1α were positively correlated with the differentiation degree of HCC and TNM stage(P ＜0.01, P ＜0.05).The expression of ki-67 was positively correlated with the differentiation degree of HCC.There was a significant positive correlation between the expressions of Glut1 and HIF-1α in HCC (r1 =0.553, P ＜0.05), the expressions of Glut1 and HIF-1α were positively correlated with ki-67(r2 =0.560,r3 =0.613, P ＜0.05).ConclusionsGlut1, HIF-1α and ki-67 may play a role in the tumorigenesis and progression of HCC in some degree.Combined detection of Glut1, HIF-1α and Ki-67 may be helpful to judge the degree of malignancy and potential metastasis and evaluate the prognosis.

Objective To investigate the effect of sertraline on the viability and the expression of tyrosine hydroxylase (TH) and phosphorylated ERK1/2 in NGF-induced rat pheochromocytoma (PC12) cells.Methods NGF-induced PC12 cells were pretreated or directly treated with different concentrations of sertraline for 24 or 48 hours and the pretreated groups were then subjected to serum withdrawal condition. Then cell viability was determined by the cell counting Kit-8 (CCK-8). The expression of Tyrosine hydroxylase (TH) and pERK1/2in NGF-induced PC12 cells was determined by immunohistochemistry and western blot respectively. Results The viability of NGF-induced PC12 was improved after administration with sertra]ine. After 24h sertraline administration, the cells activity of PC 12 cells at 20μM ( 1.32 ± 0. 11 ) , 10μM ( 1. 17 ± 0.05 ) of direct effect, and 20μM ( 1.15 ±0.11 ) of protect effect increased dramatically as compared with control group. But high dose ( 50μM )sertraline express high toxic effect to PC12 cells. The expression of TH was increased by sertraline 20 μM at both 24h(ratio of TH/β-actin = 1.27 ±0.05) and 48h(ratio of TH/β-actin = 1. 23 ±0.08) compare with control group,and the expression of pERK1/2 also increased dramatically by sertraline 20 μM at both 24h (ratio of (pERK1/2)/β-actin = 1.41±0.05) and 48h( ratio of (pERK1/2)/β-actin = 1.40 ±0.06) compare with control group(P＜0. 01, P ＜ 0. 05). Immunohistochemistry showed similar results. Conclusion These data suggest that the neuroprotective effect of sertraline may play an important role in depression therapy, and this effect might be mediated by TH and pERK1/2 up-regulation.

Objective To develop a simple and effective method for monitoring cortical ischemia after temporary occlusion of the parent arteries during anterior circulation intracranial aneurysm surgery. Methods Fifty-two patients with anterior circulation aneurysm (58 aneurysms) received craniotomy from April to November 2008, and at the same time,cortical electroencephalograpby (EEG) and scalp EEG were monitored during the surgery.According to the international 10/20 electrode placement system, scalp electrodes were placed on O1, O2, P3, P4, T5, and T6 for monitoring the changes in the depth of anesthesia. A cortical strip electrode was placed on the cortical surface supplied by the artery that was possibly blocked during the operation, which was used to monitor the possible cortical ischemia. For patients who had cortical EEG suppression after the temporary occlusion of the parent arteries Were compared with the changes of scalp EEG. Whether there were ischemic events in the corresponding supply territory after vascular occlusion were observed after surgery. Results Of the 58 aneurysms, 40 aneurysms and 41 major arteries were occluded temporarily. After being occluded temporarily in 19 arteries of 18 patients, cortical EEG changed significantly,while scalp EEG did not change significantly. Only 9 patients had ischemic events in the corresponding supply territories after the occlusion in the cortical EEG significant change group. The changes in the depth of anesthesia had the consistent impact on cortical and scalp EEG. Conelusions Simultaneous monitoring of cortical and scalp EEG is a simple and effective method for monitoring cortical ischemia during anterior circulation intracranial aneurysm surgery, and may effectively identify the effect of anesthesia on EEG.