Over the last decade, chronic critically ill (CCI) has emerged as an epidemic in intensive care unit (ICU) survivors worldwide. Advances in ICU technology and implementation of care bundles has significantly decreased early deaths of critically ill patients, and have allowed them to survive previously lethal multiple organ failure (MOF). However, more and more survivors leave persistent low grade organ dysfunctions, depend on continues organ support, need to stay in ICU, and become CCI patients. These patients experience a persistent immune dysregulation with persistent inflammation, immunosuppression, and catabolic syndrome. Therefore, malnutrition is an important feature of patients with CCI, and nutritional support is a crucial part of their treatment. The main strategies of nutritional support are as follows: providing sufficient calories and proteins with appropriate anabolic agents to promote anabolic metabolism, using immunomodulators to improve immune suppression and inflammatory responses, and supplementing micronutrients to enhance metabolic support. In this review, the nutritional assessment, calorie assessment, protein assessment and other nutrient supplementation (such as β blocker, testosterone and oxandrolone, immunonutrition, vitamins) of CCI patients were reviewed, so as to provide reference for the treatment of CCI.

Background Retina fixed flat-mount perfused by Evans blue (EB) is a common method for the evaluation of blood-retinal barrier (BRB).However,previous method is inconvenient for some laboratories because the retinal specimen can not be observed by gereral microscope rather than confocal laser scanning microscope after the fixation.Objective This study was to modify the preparing way of flat-mounted retina in order to obtain transparent specimen for the observation of rat retinal vessels and the evaluation of leakage under the ordinary fluorescence microscope.Methods Forty male SD rats were divided into the control group,diabetes mellitus (DM) 1-month group,DM 3-month group and DM 6-month group according to the random number table.Streptozotocinum (STZ) of 2％ dissolved in 0.05 mmol/L sodium citrate-hydrochloric acid buffer was intraperitoneally injected in SD rats to establish DM models,and the equal volume of solvent was injected in the same way in the control rats.One month,three months and six months after injection,EB of 30 g/L was injected via rat femoral vein in the dose of 45 mg/kg.Fifteen minutes after injection of EB,the rats were sacrificed and the retinas were isolated and cut radially to prepare the flat-mounted retinas in PBS immediately and then were dried till the specimens were transparent.The specimens were examined under the fluorescence microscope.The percentage of EB leakage was quantitatively calculated by IPP 6.0 software.All procedures were performed following approval of the institutional animal care and use committee of Tianjin Medical University.Results The retina morphology was normal in the control group,and EB filled the vessels,exhibiting the red fluorescence under the fluorescence microscope.Compared with the control group,retinal background fluorescence was enhanced slightly in the DM 1-month group,and focal leakage of the EB from capillaries and focal dilated vessels were found in the DM 3-month group,further,vascular caliber inequality,retinal hypoperfusion area and a larger number of hyperfluorescence areas were seen in the DM 6-month group.The percentage of leakage area was (0.05 ±0.02) ％,(0.27 ±0.06) ％,(1.17 ±0.18)％ and (4.77 ±0.66)％ in the control group,DM 1-month group,DM 3-month group and DM 6-month group,respectively,showing a significant difference among the four groups (F =795.800,P＜0.001),and the leakage area was obviously larger in the DM 3-month group and DM 6-month group than that in thecontrol group (q'=10.338,q'=43.475,both at P＜0.001).Conclusions Modified EB-perfused retinal wholemount method is easy and helpful for clear visualization of retinal vessel leakage induced by BRB breakdown in the diabetic rats under the common fluorescence microscope.

Objective: To investigate the effect of enolase 1 (ENO1) expression on proliferation, apoptosis and migration of lung cancer PC14 cells. Methods: ENO1 over-expression vector-pcDNA3.1/ENO1 was constructed and transfected into PC14 cells at logarithmic growth phase with liposome LipofectamineTM 2000. G418 was used to screen PC14 cells that stably expressing ENO1. The effects of ENO1 over-expression on proliferation, migration and apoptosis of PC14 cells were detected by CCK-8 method, scratch-healing assay and flow cytometry, respectively. Results: The ENO1 over-expression cell model was successfully constructed. Compared with PC14-vehicle and wild-type PC14 cells, the mRNA and protein expression levels of ENO1 in PC14-ENO1 cells were significantly elevated (all P<0.05), and the proliferation of PC14-ENO1 cells was significantly increased (all P<0.05). The relative mobility of PC14ENO1 cells was significantly higher than that of pcDNA3.1-vehicle cells and wild-type PC14 cells ([13.26±1.13]% vs [8.46±1.11]%, [7.86±1.00]%, both P<0.05). There was no significant difference in apoptotic rate among PC14-ENO1, PC14-vehicle and PC14 cells (all P> 0.05) Conclusion: Over-expression of ENO1 promotes proliferation and migration of lung cancer PC14 cells.

Objective:To establish a 180-day mortality predictive score based on frailty syndrome in elderly sepsis patients [elderly sepsis score (ESS)].Methods:A prospective study for sepsis patients aged 60 years and above who were admitted to a medical intensive care unit of the General Hospital of Southern Theatre Command from January 1st, 2018 to December 31st, 2018 was conducted. Univariate analysis was performed on 19 independent variables including gender, age, body mass index (BMI), tumor, charlson comorbidity index (CCI), activity of daily living (ADL), instrumental activity of daily living (IADL), mini-mental state examination (MMSE), geriatric depression scale (GDS), clinical frail scale (CFS), sequential organ failure assessment (SOFA), Glasgow coma scale (GCS), acute physiology and chronic health evaluation (APACHEⅡ, APACHEⅣ), modified NUTRIC score (MNS), multiple drug resistance (MDR), mechanical ventilation (MV), continuous renal replacement therapy (CRRT) and palliative care. Continuous independent variables were converted into classified variables. Multivariate binary regression analysis of risk factors was conducted to screen independent risk factors which affecting 180-day mortality in elderly sepsis patients. Then a 180-day mortality predictive score was established, and the discrimination of the mortality of patients using CFS, SOFA, GCS, APACHEⅡ, APACHEⅣ, MNS scores were compared.Results:A total of 257 patients were enrolled, with a 180-day mortality of 60.7%. Univariate analysis showed that age, tumor, CCI, ADL, IADL, MMSE, CFS, SOFA, GCS, APACHEⅡ, APACHEⅣ, MNS, MDR, MV, CRRT, palliative care were risk factors of 180-day mortality in elderly sepsis patients [age: odds ratio ( OR) = 1.027, 95% confidence interval (95% CI) was 1.005-1.050, P = 0.018; tumor: OR =2.001, 95% CI was 1.022-3.920, P = 0.043; CCI: OR = 1.193, 95% CI was 1.064-1.339, P = 0.003; ADL: OR = 0.851, 95% CI was 0.772-0.940, P = 0.001; IADL: OR = 0.894, 95% CI was 0.826-0.967, P = 0.005; MMSE: OR = 0.962, 95% CI was 0.937-0.988, P = 0.004; CFS: OR = 1.303, 95% CI was 1.089-1.558, P = 0.004; SOFA: OR = 1.112, 95% CI was 1.038-1.191, P = 0.003; GCS: OR = 0.918, 95% CI was 0.863-0.977, P = 0.007; APACHEⅡ: OR = 1.098, 95% CI was 1.053-1.145, P < 0.001; APACHEⅣ: OR = 1.032, 95% CI was 1.020-1.044, P < 0.001; MNS: OR = 1.315, 95% CI was 1.159-1.493, P < 0.001; MDR: OR = 2.029, 95% CI was 1.197-3.437, P = 0.009; MV: OR = 6.408, 95% CI was 3.480-11.798, P < 0.001, CRRT: OR = 2.744, 95% CI was 1.529-4.923, P = 0.001, palliative care: OR = 5.760, 95% CI was 2.177-15.245, P < 0.001]. By binary regression analysis, CFS stratification ( OR = 1.934, 95% CI was 1.267-2.953, P = 0.002), MV ( OR = 4.531, 95% CI was 2.376-8.644, P < 0.001), CRRT ( OR = 2.471, 95% CI was 1.285-4.752, P = 0.007), palliative care ( OR = 6.169, 95% CI was 2.173-17.515, P = 0.001) were independent risk factors of 180-day mortality in elderly patients with sepsis. The model of "ESS = 0.660×CFS stratification+1.511×MV+0.905×CRRT+1.820×palliative care" was established. Receiver operating characteristic curve (ROC curve) analysis showed that the area under the ROC curve (AUC) for predicting 180-day mortality by ESS was 0.785 (95% CI was 0.730-0.834, P < 0.001). When the best cut-off value was 2.2 points, its sensitivity was 78.9%, specificity was 70.3%, the positive predictive value was 80.4%, and the negative predictive value was 68.3%. Simplified ESS was defined as "0.5×CFS stratification+1.5×MV+1×CRRT+2×palliative care". ROC curve analysis showed that AUC for predicting 180-day mortality by simplified ESS was 0.784 (95% CI was 0.729-0.833, P < 0.001). When the best cut-off value was 2.0 points, sensitivity was 76.9%, specificity was 70.3%, the positive predictive value was 80.0%, and the negative predictive value was 66.4%. Compared with CFS, SOFA, GCS, APACHEⅡ, APACHEⅣ and MNS, ESS had a significant difference in discriminating 180-day mortality in elderly patients with sepsis (AUC was 0.785 vs. 0.607, 0.607, 0.600, 0.664, 0.702, 0.657, 95% CI: 0.730-0.734 vs. 0.537-0.678, 0.537-0.677, 0.529-0.671, 0.598-0.730, 0.638-0.766, 0.590-0.725, all P < 0.05). Conclusions:CFS, MV, CRRT, and palliative care are independent risk factors of 180-day mortality in elderly patients with sepsis. We established ESS based on these risk factors. The ESS model has good discrimination and can be used as a reference and assessment tool for prediction and treatment guidance in elderly patients with sepsis.

OBJECTIVE:To put forward relevant suggestions for formulating and implementing medical insurance budget impact analysis(BIA)research guideline in China. METHODS:The medical insurance BIA guidelines or relevant documents were retrieved from ISPOR,Canada and the United States. Their similarities and differences were compared. The experiences of them were summarized in respects of research framework,data source and report format of medical insurance BIA. RESULTS &CONCLUSIONS:ISPOR,Canada,USA and other guidelines or relevant documents have some similarities in normalization requirements of research perspectives,target population calculation based on natural reason and epidemiological data,3-5 years as the research time limit and some other respects. But the calculation of additional costs as drug price addition,distribution fee should be specifically adjusted according to the unique characteristics of the health systems of countries or regions. Based on the actual conditions of our own health care system,our country can draw up the guidelines for medical insurance BIA impact analysis,which contain model design,research perspective,target population,current use of intervention measures,prediction on the effects of new intervention measure introduction on the market,cost,time range,discount and uncertainty analysis of current and new interventions,situational analysis and verification,so as to better play its role in the formulation and adjustment of medical insurance catalogues and in drug price negotiations.

Objective To observe the effect of Yijing Fang,a TCM compound,on the expression of junctional adhesion molecules (JSM)and zonula occludens (ZO)in testicular sertoli cells of infertile rats with kidney-yang deficiency pattern induced by adenine.Methods Altogether 48 healthy male Wistar rats of SPF grade,two-month-old,were divided according to random number table,into blank group, model group,Fufang Xuanju group and Yijing Fang group(n =12 each).After modeling,the rats were intragastrically administered by corresponding medicines for consecutive 20 days.Then the rats were sac-rificed and their testis and epididymis were resected.The sperm density and sperm activity were calcula-ted using Computer Assisted Sperm Analysis System,and the expression of JAMand ZO in testicular ser-toli cells were measured using immunohistochemical method.Results Compared with the normal group, the sperm density,sperm activity rate and expression of JAMand ZO declined significantly(P <0.01). Compared with the model group,the sperm density and the sperm activity rate increased in Fufang Xuan-Ju group and Yijing Fang group remarkably (P <0.05),while expression of JAM and ZO increased in Yijing Fang group((P <0.05 ).Conclusion Yijing Fang can repair the damage of testicular sertoli cells of kidney-yang deficiency pattern rats induce by adenine,via regulating the expression of JAM and ZO.

Objective To evaluate the effect of clopidogrel premedication on in-hospital major adverse cardiovascular and cerebral events (MACCE) and bleeding outcomes before coronary artery bypass graft surgery (CABG). Method A total of 2021 patients who underwent CABG from July 2003 to September 2005 were divided into either clopidogrel ( n = 479) or no clopidogrel (1542) group before CABG. Patients with clopidogrel administration ( n = 479) were subdivided into < 5 d ( n = 154) ,5～7d(n = 183)and >7d(n = 142) group according to timing of clopidogrel withdrawal before surgery. In-hospital MACCE and perioperative bleeding outcomes were analyzed among groups. Results Patients who took clopidogrel before surgery had nonsignificantly rates of bleeding and in-hospital MACCE compared with those patients not administered clopidogrel. No differences were found about the incidence of total bleeding,minor bleeding,transfusions of red blood cells,fresh frozen plasma,whole blood and in-hosptial MACCE among three subgroups.The < 5 d group had higher incidence of major bleeding and more platelets transfusions than 5 ～ 7 d [47.8% vs. 31.9%,P < 0.017; (0.08 ±0.38) U vs. (0.00±0.00) U,P <0.017,respectively]and >7 d group [47.8% vs. 20.3%,P <0.017; (0.08±0.38) U vs. (0.00±0.00) U,P <0.017,respectively). However,there were no significant differences between 5 ～ 7 d and > 7 d group ( P > 0.05). Conclusions Gopidogrel administration before CABG does not increase the incidence of in-hospital MACCE events. However,the perioperative risk of bleeding will rise if the patients withhold clopidogrel less than five days before surgery.

Objective To assess the effect of anemia on long-term outcomes in patients with acute coronary syndrome(ACS) undergoing pereutaneous coronary intervention(PCI). Methods In 3136 patients presenting with ACS,636 patients were anemic. The clinical features, mortality and major cardiocerebral events including non-fatal acute myocardial infaret,revascularization and non-fatal cerebral stroke were compared in patients with or without anemia. The average follow-up period was 550 days. Results Anemic patients were older and had a higher percentage of comorbidities compared with nonanemic cohorts. Compared with nonanemic patients, anemic patients had higher mortality (4.7% versus 1.5% ,P <0. 001) and a higher major adverse end point events,including nonfatalmyocardial infarction, stroke and revaseularization (14.2% versus 11.0%, P = 0.032). After adjustment for comorbidities, anemia was associated with a higher risk of mortality after percutaneous coronary intervention (adjusted hazard rate ratioRR2. 166 ;95% CI 1. 298-3. 612 ;P =0.003). Conclusion Anemia before PCI is an independent factor for predicting the long-term mortality of ACS.

Objective To identify the electrophysiological charateristics and cause of ridge gap related reentry after MI ablation in atrial ifbrillation patients. Methods Activation and entrainment mapping was performed in 82 redo cases for OAT recurrence in whom MI was ablated during the index produre. Once ridge gap related reentry was conifrmed, detailed mapping was performed in MI and ridge region. In addition, in 36 cases undergoing MI ablation and fulfilling criterion for bidirectional block, differential pacing was repeated at the ridge to identify a ridge gap. Results Out of 82 redo cases for OAT recurrence in whom MI was ablated during the index produre, 7 (8.5%) was found to be ridge gap related reentry. TCL was (247.9±19.2) ms, and the left atrial endocardial activation time was (145.4±17.7) ms, accounting for (58.5±3.2)%of TCL. However, wide double potential was recorded along the previous ablated MI line where PPI was (34.3±6.6) ms longer than TCL, while PPI was signiifcantly shorter at the ridge[PPI-TCL (11.4±3.9) ms, P<0.001]. Tachycardia was terminated at the ridge in 6 cases and at the corresponding site in coronary sinus in 1 case. No recurrence was found during follow-up for (11.1±4.5) months. In addition, in 36 patients undergoing MI ablation in whom criterion of bi-directional block was fuliflled, conduction gap located at the ridge was found in 5 (13.9%) cases. Conclusions MI ridge gap related reentry is a distinctive OAT, in which the ridge was used as the critical isthmus, whereas the previous ablated MI line is not part of the reentry. MI pseudo-block due to the ridge gap may lead to this type of recurrent tachycardia.

Objective Identify the mechanism of induced atrial arrhythmias after pulmonary vein isolation (PVI) in patients with paroxysmal atrial ifbrillation(PAF), and investigate its long-term prognosis. Methods All patients with PAF undergoing PVI and induction test afterwards between Feburary 2010 and October 2010 were included. The induction protocol was rapid pacing initiated at cycle length of 250 ms with progressive shortening in a decrement of 10 ms down to 180 ms or refractoriness. Isoproterenol of 2-4μg/min was administrated as well. Inducibility was deifned as induction of atrial arrhythmia lasting >1 min. The mechanism of induced tachycardia was identiifed by activation mapping and entrainment mapping under the guidance of CARTO system. All patients were followed up by 36 months. Results Forty-nine atrial tachycardia were induced in 39 (19.7%) patients, including 35 organized atrial tachycardia (OAT) and 14 atrial ifbrillation (AF). The LA diameter was signiifcantly larger in inducible group than non-inducible group (39.5±6.6 mm vs. 36.7±5.2 mm, P=0.004). Macroreentry was the most common mechanism in induced OATs (28, 80.0%), and mitral isthmus was the most common critical site (20, 40.8%), followed by cavo-tricuspid isthmus (12, 24.5%), PV (6, 12.2%), LA septum (4, 8.2%), superior vena cava (3, 6.1%) and LA roof (1, 2.0%). Conclusions The most common mechanism of induced tachycardia by IV isoproterenol and rapid pacing is MI and CTI dependent after PVI in PAF patients, which can be succssefully eliminated by liner ablation, not increasing long-term recurrence rate.