Objective To investigate the changes of Notch signaling pathway activity in human pancreatic cancer cell lines (SW1990, BxPC3 )after gemcitabine induction, and to study its relationship with pancreatic cancer resistant to gemcitabine chemotherapy. Methods The pancreatic cancer cell lines SW1990 and BxPC3 were cultured with different concentrations of gemcitabine for 48 hours. The Notch signaling pathway receptors ( Notch1, Notch2, Notch3, Notch4), ligands (Jagged1, Jagged2) and downstream target Hesl mRNAs expression were detected by quantitative real-time PCR (Q-PCR). Protein levels of Hes1 were determined by Western blotting. Results After treatment with 2 μmol/L gemcitabine for 48 hours, the expression of Notch1, Notch2, Notch3, Jagged1, Jagged2 and Hes1 mRNAs in SW1990 cells were 8.26 ±0.48, 39.12 ±4.87, 0.84 ±0.06, 105.8 ± 17.92, 6.59 ±0.32 and 17.30 ±2.96, which were significantly elevated when compared with those without gemcitabine treatment ( 1.02 ± 0. 15, 15.25 ± 1.28, 0. 12 ± 0.02,32.66 ± 1.98, 1.88 ± 0.29 and 5.02 ± 0.64, P < 0.05 or P < 0. 01 ); the expression in BxPC3 cells was 7.87 ±0.59, 109.4 ± 10.98, 0.74 ±0.19, 62.73 ± 13.50, 2.09 ±0.16 and 15.38 ± 1.06, which were significantly elevated when compared with those without gemcitabine treatment ( 1.14 ±0.43, 58.96 ±2.63,0.10 ± 0.02, 16.95 ± 3.79, 0.98 ± 0.02 and 2.04 ± 0.16, P < 0.05 or P < 0.01 ). The expressions of Hes1protein in SW1990 cells after 1, 2 μmol/L gemcitabine treatment for 48 h were 0.30 ±0.03, 0.42 ±0.03;and the expressions in BxPC3 cells were 0.33 ± 0.02, 0.45 ± 0.03, which were significantly increased when compared with those without gemcitabine treatment (0.13 ± 0.01, F = 33.71,0.09 ± 0.02, F = 38.54, P <0.01 ). Conclusions The Notch signaling pathway is significantly activated in pancreatic cancer cells SW1990 and BxPC3 by gemcitabine, which may be one of the mechanisms of chemoresistance.

AIM: To investigate the changes of small intestine villus and sublingual microcirculation perfusion in the rabbits during endotoxic shock by sidestream dark-field imaging (SDF) after resuscitation to a mean arterial pressure (MAP) level.METHODS: New Zealand white rabbits (n=60) were randomly divided into 2 groups (group of villus and group of sublingua).The fistula operation of ileum was performed.Lipopolysaccharide was injected to establish endotoxic shock model, and fluid resuscitation (lactated Ringer's solution, 30 mL·kg-1·h-1) was given to maitain the MAP of the animals to 80 mmHg.Continuous norepinephrine was intravenously injected at 0.5~1 μg·kg-1·min-1 only if fluid therapy did not maintain the MAP level.The changes of microcirculatory perfusion indexes in small intestine villus and sublingual tissues such as vessels per villus (VV), microvascular flow index (MFI), proportion of perfused villi (PPVi), villus border score, villus vessel score, total vessel density (TVD), perfused vessel density (PVD) and proportion of perfused vessels (PPVe) were continuously observed and recorded by SDF before shock, during shock and after fluid resuscitation.RESULTS: MFI and PPVi in small intestine villus, and MFI, PPVe, TVD and PVD in sublingual tissues were significantly decreased after shock (P<0.01).Compared with MFI in sublingual microcirculation, MFI in villus was significantly decreased (P<0.01).MFI and PPVi in small intestine villus, and MFI, PPVe, TVD and PVD in sublingual tissues were improved after recovered to the target MAP by fluid resuscitation (P<0.05).However, MFI in small intestine villus was significantly lower than that in sublingual tissues after fluid resuscitation (P<0.01).CONCLUSION: The difference between small intestine villus and sublingual microcirculation perfusion during endotoxic shock is observed.The descent degree of microcirculation perfusion in small intestine villus is larger than that in sublingual tissues after shock, and the recovery degree of small intestine villus microcirculation is lower than that of sublingual microcirculation afer fluid resuscitation.

Objective Changes of small intestine villus microcirculation perfusion in sidestream dark-field (SDF) imaging in the rabbits during endotoxic shock after fluid resuscitation with different target mean arterial pressure (MAP), and evaluation of feasibility of monitoring small intestine villus microcirculation by SDF were studied. Methods Sixty standard New Zealand white rabbits were randomly divided into two groups: low target MAP group (group A, n = 30) and high target MAP group (group B,n = 30). Fistula operation of ileum was madein vitro, and lipopolysaccharide (LPS, 2 mg/kg) was injected to establish endotoxic shock model. Group A was administered with the lower dose fluid resuscitation (lactated Ringer solution, 20 mL·kg-1·h-1) for target MAP of 65 mmHg (1 mmHg =0.133 kPa); group B was administered with the higher dose fluid resuscitation (lactated Ringer solution, 30 mL·kg-1·h-1) for MAP of 80 mmHg. Continuous norepinephrine intravenous injection (0.5-1.0μg·kg-1·min-1) was administered only after fluid therapy couldn't reach the target MAP. The changes of small intestine villus microcirculation perfusion indexes such as vessels per villus (VV), proportion of perfused villi (PPV), microvascular flow index (MFI), borders of villus score (BVS), vessels villus score (VVS) were continuously observed and recorded before the shock, during the shock and after fluid resuscitation using SDF imaging. The differences of microcirculation perfusion were compared between two groups using the specific parameter evaluation system to determine severity of villi microcirculation and injury scores at different stages.Results VV and borders of villus were clear and contact before shock in two groups. After shock, VV, PPV were significantly decreased in both two groups, the borders of villus were destroyed, MFI, BVS, VVS and the total score of villi injury microcirculation were obviously and severely decreased. Partial blood flow of villous capillaries after fluid resuscitation was recovered in two groups, but the perfusion of some region was un-balanced with the outworn borders of villus. VV were rose as compared before and after fluid resuscitation in groups A and B (vessels: 1.21±0.22 vs. 0.81±0.12, 1.54±0.28 vs. 0.79±0.13), and PPV [(31±4)% vs. (12±2)%, (38±5)% vs. (13±3)%], MFI (1.55±0.09 vs. 1.09±0.03, 1.97±0.11 vs. 1.05±0.03), VVS (points: 1.22±0.08 vs. 0.89±0.02, 2.06±0.15 vs. 0.90±0.02) and the sum of MFI, BVS, VVS (3.70±0.19 vs. 2.85±0.07, 5.01±0.29 vs. 2.88±0.08) were significant rose (allP< 0.05). The recovery of group B was better than that of group A, and the injury score was reduced. But BVS were not increased in both groups compared with before and after shock (points: 0.93±0.05 vs. 0.87±0.03, 0.98±0.09 vs. 0.93±0.05, bothP > 0.05).Conclusions For the small intestine villus microcirculation perfusion, the higher target MAP (80 mmHg) after fluid resuscitation or/and vasoconstrictor drugs usage were probably better than the relatively lower target MAP (65 mmHg) during endotoxic shock. SDF imaging is a very promising technique for intestinal villi microcirculatory visualization and assessment.

Objective To compare the value of bedside index for severity in acute pancreatitis (BISAP),Ranson score and Balthazar computed tomography severity index (CTSI) in predicting the severity and prognosis of acute pancreatitis (AP).Methods From 2005 to 2011 in Shanghai,the clinical data of 1004 AP cases from seven hospitals was collected and retrospectively analyzed.The value of BISAP score,Ranson score and Balthazar CTSI in predicting the severity and prognosis of AP were assessed with receiver operator characteristic (ROC) curve.Results Among 1004 patients,the main cause of AP was biliary disease (580 cases),about 57.77％.The incidence of pancreatic necrosis,mortality and SAP increased along with BISAP score.The risk of pancreatic necrosis in patients with CTSI ≥ three was significantly higher than that of ＜ three.The risk of pancreatic necrosis and SAP in patients with BISAP score ≥ two was significantly higher than that of ＜ two (OR:4.93,95％CI 3.62-6.70; OR 2.62,95％CI 1.59-4.31,respectively).There was no significant difference in the accuracy of predicting the progression and mortality of AP among these three score systems.However the sensitivity of BISAP score (OR:61.54,95％CI 35.09-87.99) in predicting the progression and mortality of AP was better than that of Ranson (OR:46.15,95 ％ CI 19.05-73.25) and CTSI (OR:46.15,95％CI 19.05-73.25).Conclusions BISAP score is easy to perform and when combined with CTSI,it helps to make the diagnosis and classification of AP in time,predict the prognosis accurately.Compared with Ranson score,BISAP score has higher clinical value.

Objective: To investigate the effects of uncoupling protein 2 (UCP2) overexpression on mitochondrial dynamics (mitochondrial division and fusion) of sepsis myocardial injury in rats. Methods: Forty male Sprague-Dawley (SD) rats were randomly divided into four groups (n = 10): sham operation group (Sham group) using normal saline instead of transfection and simulating cecal ligation and perforation (CLP); CLP group using normal saline instead of transfection, performing CLP to induce sepsis; adeno-associated virus (AAV) group using CLP after myocardial transfection with empty virus; UCP2 overexpression group (UCP2 group) CLP was performed 3 weeks after AAV-UCP2 (1×10¹⁵ vg/L, a total of 60 μL) myocardial transfection. The rats in each group were examined by echocardiography at 24 hours after the CLP, and then the rats were sacrificed immediately to harvest myocardial tissue. Myocardial ultrastructural changes were observed under the electron microscope, the expression of regulatory proteins related to myocardial mitochondrial dynamics [optic atrophy 1 (Opa1), dynamin-related protein 1 (Drp1) and fission 1 (Fis1)] were detected by Western Blot, and the level of mitochondrial adenosine triphosphate (ATP) production was detected by chemiluminescence. Results: ①The echocardiographic results showed that there was no significant difference in left ventricular mass (LVM) and stroke volume (SV). Compared with Sham group, left ventricular diastolic anterior wall thickness (LVAWd), left ventricular systolic anterior wall thickness (LVAWs), left ventricular diastolic posterior wall thickness (LVPWd), left ventricular systolic posterior wall thickness (LVPWs), left ventricular ejection fraction (LVEF) and left ventricular short axis shortening rate (LVFS) were significantly increased in CLP group and AAV group, while left ventricular systolic diameter (LVEDs), left ventricular diastolic diameter (LVEDd), left ventricular end-systolic volume (LVESV), and left ventricular end-diastolic volume (LVEDV) were significantly decreased. Compared with CLP group and AAV group, LVAWs, LVEF, LVFS were significantly decreased in UCP2 group, and LVEDs, LVEDV and LVESV were significantly increased [LVAWs (mm): 3.82±0.42 vs. 4.34±0.30, 4.44±0.12; LVEF: 0.921±0.038 vs. 0.979±0.019, 0.991±0.010; LVFS: (65.33±6.56)% vs. (80.11±8.23)%, (85.31±6.11)%; LVEDs (mm): 1.81±0.36 vs. 0.89±0.54, 0.60±0.17; LVEDV (μL): 137.09±50.05 vs. 89.72±53.04, 85.42±40.99; LVESV (μL): 10.48±4.59 vs. 2.48±3.52, 2.58±2.50, all P < 0.05]. ② Electron microscope showed that the structure of myocardial fibers in the Sham group was clear and aligned with complete intervertebral disc and mitochondrial structure, no damage to mitochondrial membranes, and tight arrangement of cristae. In CLP group and AAV group, muscle fiber breakage, sarcoplasmic reticulum expansion, severe mitochondrial swelling and even cristage structure disorder were observed. In the UCP2 group, only myocardial fiber edema was observed, and the muscle fiber structure was more complete than that of Sham group and AAV group. The mitochondria were slightly swollen and the cristae were intact.③ Western Blot showed that there was no significant difference in the expression of Opa1 and Fis1 in the four groups. The expression of Drp1 in CLP group and AAV group were significantly higher than that in Sham group. The expression of Drp1 in UCP2 group was significantly lower than that in CLP group and AAV group (Drp1/β-actin: 1.01±0.03 vs. 1.39±0.03, 1.49±0.03, both P < 0.05).④ The results of immunofluorescence showed that the ATP content of CLP group and AAV group were significantly lower than that of Sham group; the ATP content of UCP2 group was significantly higher than that of CLP group and AAV group (μmol/L: 1.99±0.15 vs. 1.10±0.17, 1.13±0.19, both P < 0.05). Conclusion UCP2 overexpression can significantly improve the systemic systolic function of myocardium in sepsis rats, protect myocardial mitochondrial ultrastructure, inhibit mitochondrial division, and improve mitochondrial ATP synthesis.

Barrett’s esophagus (BE) is the recognized precancerous lesion and risk factor for esophageal adenocarcinoma (EAC), and has a high miss diagnosis rate and low survival rate when malignantly transformed into EAC, moreover, there are only limited monitoring method and treatment. Therefore, the screening of biomarkers is highly expected, especially the risk stratification biomarkers related to the progression of malignant transformation of BE. Such biomarkers can help to determine early, quickly and accurately the disease process, and guide the stratified management and precise treatment of BE, reduce the malignancy rate and mortality. This article focused on the dynamic evolutionary process of intra‑tumor heterogeneity, and reviewed the current status and challenges of research on BE biomarkers in risk stratification from the genetics, epigenetics and serology perspectives.

OBJECTIVE: To explore the diagnosis and treatment of acute cerebral infarction following extracorporeal membrane oxygenation (ECMO) therapy in patients with cardiogenic shock to review the literature. METHODS: The clinical data of two patients with cardiogenic shock treated with veno-arterial ECMO (VA-ECMO) complicated with acute cerebral infarction admitted to department of intensive care unit (ICU) of Affiliated Hospital of Guizhou Medical University were retrospectively analyzed and the treatment experience was shared. RESULTS: Case 1 was a 46-year-old male patient who was admitted to the hospital on September 16, 2021, due to "repeated chest tightness, shortness of breath, syncope for 2+ years, and worsened for 15 days. Coronary artery angiography showed 3-vessel coronary artery disease lesions. On October 15, 2021, coronary artery bypass grafting (CABG), pericardial fenestration and drainage, thoracic closed drainage, femoral bypass, thoracotomy exploration, and sternal internal fixation were performed under support of extracorporeal circulation. After surgery, the heart rate was 180-200 bpm, the blood pressure could not be maintained, and the improvement was not obvious after active drug treatment. The right femoral artery and femoral vein were intubated, VA-ECMO support treatment was performed, and the patient was transferred to the ICU. Intra-aortic balloon pump (IABP) was treated on the day of transfer because the circulation could not be maintained. Due to acute cerebral infarction in the left hemisphere and right parieto-occipital lobe, subfalcine herniation, tentorial herniation, the patient ultimately died after withdrawing from ECMO. Case 2 was a 43-year-old male patient who was admitted to the hospital on June 29, 2021, with "fever for 8 days and vomiting for 4 days". Bedside ultrasound showed cardiac enlargement and diffuse wall motion reduction in the left and right ventricles. On June 30, 2021, the patient underwent catheterization through the right femoral artery and femoral vein, VA-ECMO support, and was transferred to ICU for treatment. Acute cerebral infarction on both sides of the cerebellum occurred, and after treatment, the patient was discharged with mild impairment of daily living ability. CONCLUSIONS Strengthen monitoring of anticoagulation; regular neurological examination of patients undergoing ECMO therapy; ECMO under light sedation or awake can be performed if the condition permitsif the condition permits, perform light sedation or awake ECMO, which helpful for the early detection of nervous system injury.
Male ; Humans ; Middle Aged ; Adult ; Shock, Cardiogenic/therapy* ; Extracorporeal Membrane Oxygenation ; Retrospective Studies ; Coronary Artery Bypass/adverse effects* ; Cerebral Infarction/therapy*