Objective To summarize the clinical features of patients suffering from pulmonary embolism (PE) with D-dimer ＜ 0. 5 μg/mL in order to raise the diagnostic accuracy and reduce the mortality rate of PE. MethodsD-dimer-negative patients with suspected PE were admitted from January 2006 through December 2009. A comparison of clinical features including clinical manifestations, vital signs, laboratory and ancillary findings between 16 patients finally diagnosed PE and 41 patients without PE. ResultsCompared with patients without PE, the D-dimer-negative patients with PE usually had past history of venous thromembolism (VTE) or recent surgery. The symptoms of chest tightness, acute dyspnea, tachypnea, lower extremity edema and typical S I QⅢTⅢ changes of ECG were more often occurred in patients with PE than those in patients without PE of control group. ConclusionsD-dimer test is a good screening test for acute PE because its negative predictive value is high, but when the patients have acute dyspnea, lower extremity edema, previous history of VTE or/and recent surgery and ECG SI QⅢ TⅢ changes, even if D -dimer ＜ 0. 5 μg/mL, clinicians also should pay attention to and if necessary, further tests should be considered to confirm the diagnosis of PE.

Acute-on-chronic liver failure is a common form of liver failure characterized by complicated clinical manifestations and high mortality.The pathophysiology of ACLF is still unclear.More efficacious treatments are based mainly on a better understanding of the pathophysiology of ACLF.The advances in the pathophysiology of ACLF have been extremely encouraging in the last few years.In this article,we reviewed the progress of ACLF,its definition,etiology and,pathophysiology.

Objective To investigate the expressions and correlation between Survivin and PTEN proteins in astrocytoma.Methods The expressions of Survivin and PTEN proteins were examined by immunohistrochemistry in astrocytoma specimens from 65 patients with astrocytoma.Results The positive expression rate of Survivin in astrocytoma grade Ⅱ was significantly lower than that in grade Ⅲ and grade Ⅳ(all P

Objective To investigate the role of IL-6 in the co-cultivation system of porcine hepatocytes with bone marrow mesenchymal stem cells (MSCs) in vitro.Methods Mononuclear cells were isolated from bone marrow aspirate of swines (n = 3) by 1.077 g/ml density gradient centrifugation.MSCs of passage 3 and primary hepatocytes harvested by a two-step in situ collagenase perfusion technique were inoculated in Millicell culture inserts at a seeding ratio of 1:2,and functional changes of heterotypic interactions were characterized.Levels of IL-6,TNF-α,and TGF-α were measured respectively.Results Hepatocyte viability was greater than 95%.Hepatocyte performance levels such as albumin secretion and urea synthesis were all significantly enhanced in co-culture group compared with hepatocyte homo-culture (P＜0.05).The IL-6 level also significantly increased in co-culture group (P ＜ 0.01).A significant decrease of hepatic functions was observed upon neutralization of IL-6 in co-culture.Conclusion MSC-derived IL-6 plays a key role in the up-regulation of hepatocyte functions in this co-culture.

Objective To establish porcine hepatocyte co-culture system with bone marrow mes-enchymal stem cells (MSCs) in vitro for the ideal cell source of bioartificial liver. Methods Mononu-clear cells were isolated from bone marrow aspirate extracted from the anterior superior lilac spine of swines (n=3) by 1.077 g/ml density gradient centrifugation. MSCs of passage 3 and primary hepato-cytes harvested by a two-step in situ collagenase perfusion technique were randomly distributed, and the morphological and functional changes of heterotypic interactions were characterized. Results The purity of the third passage MSCs and primary hepatoeytes was more than 90 % and 99%, respectively.Hepatocyte viability was greater than 95 %. A rapid attachment and self-organization of three-dimen-sional hepatocyte spheroids were encouraged in co-culture. Heterotypic junctions remained similar to that of hepatocytes in vivo. Hepatocyte performance levels such as albumin secretion and urea synthe-sis were all significantly enhanced in co-culture group compared with hepatocyte homo-culture (P<0.05). The best hepatic function levels were achieved on day 2 and moderately decreased in the following co-culture days. Conclusion Co-cultivation of porcine hepatocytes and MSCs may preserve hepatocyte morphology and function, which could contribute to the functional bioartificial liver.

Objective To summarize our experience with off-pump coronary artery bypass grafting (OPCAB) during previous 13 years. Methods Data from 3703 patients who underwent OPCAB between October 1996 and December 2008 were collected and analyzed in this study. Following perioperative variables were reviewed and evaluated: changes in the number of patients, demographic characteristics of patients, coexisting conditions such as hypertension, and diabetes, grafting options,numbers of grafts per patient, and postoperative complications and clinical outcomes. Patients were divided into four age subgroups: those who were less than 45 years were assigned to group 1, those who were 45 to 60 years were assigned to group 2,those who were 60 to 75 years were assigned to group 3, and those older than 75 years of age were assigned to group 4. Perioperative data, including the use of internal mammary artery and the constituent of the grafts, were collected retrospectively and analyzed. Results Three thousand and twenty-five patients were male (81.7%) and 678 were female ( 18.3% ), mean age was (61.35 ±9.38) years old. The number of patients who underwent OPCAB increased steadily over time. The mean grafts per patient were 3.3 ± 0.8. The use of left internal mammary artery and "hybrid" bypass grafts composed of vein and artery played a predominant role in this cohort (P<0.05) . During this period of 13 years, intra-aortic balloon counterpulsation (IABP)was performed in 41 patients and continuous renal replacement therapy (CRRT)was required in 12 patients. Main complications included rethoracotomy for bleeding and tamponade in 1.49% of patients, deep sternal wound infection requiring re-exploration in 1.38%, perioperative myocardial infarction in 1.03%, neurological adverse events in 0.62%, tracheotomy in 0.59%, acute renal dysfunction in 0.77%, and other complications in 0.77%. The overall in-hospital mortality was 0.7% (26 of 3703 patients). A trend toward a reduction in morbidity and mortality was shown in this study. Diseases associated with hospital mortality were cardiac sudden death, multiple organ dysfunction syndrome, low cardiac output syndrome, severe infection, extensive myocardial infarction and neurological adverse events. Conclusion The indications for OPCAB, an innovative revascularization strategy, have been expanded and the curative rate for OPCAB has been improved in recent years. Appropriate and practical grafting strategies, as well as complete perioperative management, are considered as contributors to the improved outcomes.

AIM:Toinvestigatetheeffectofnicotinicacidamide(NAA)ontheinfusiondamageofhuman umbilical cord mesenchymal stem cells ( hUC-MSCs) under the condition of instant blood-mediated inflammatory reaction ( IBMIR) .METHODS:Normal peripheral blood without anticoagulant at volume of 2.7 mL was mixed with 0.3 mL phys-iological saline (as blank group), CFSE labeled hUC-MSCs (1 ×106 cells in 0.3 mL as MSC group) and CFSE labeled hUC-MSCs (1 ×106 cells in 0.3 mL) preprocessed with NAA at concentration of 10 mmol/L for 24 h ( as MSC+NAA group) , respectively.The mixture was immediately injected into the improved Chandler Loop model, placed in 37℃water bath, and then started the peristaltic pump at the speed of 20 mL/min for 1 h.The number of CFSE labeled hUC-MSCs, platelets, white blood cells were counted and the concentration of complement C3a was measured before and after cycling, respectively.RESULTS: After 1 h circulation, the platelet dissipation rate were ( 29.96 ±10.88 )% in blank group, (77.76 ±19.29)% in MSC group all and (50.13 ±18.10)% in MSC +NAA group; and the leukocyte counts were (37.82 ±13.81)%in blank group, (64.57 ±17.08)% in MSC group and (41.52 ±17.26)% in MSC+NAA group. Compared with blank group, the differences of the dissipation rates in MSC group and MSC+NAA group all had statistical significance.The hUC-MSCs relative survival rate in MSC+NAA group was higher than that in MSC group.C3a concentra-tions in blank group, MSC group and MSC+NAA group were (206.27 ±58.10), (230.47 ±39.61) and (208.37 ± 40.66) μg/L, respectively.CONCLUSION:Co-circulating the mixture of hUC-MSCs with normal peripheral blood with-out anticoagulant in the improved Chandler Loop for 1 h depletes a large number of hUC-MSCs and blood components, and increases C3a, suggesting that this model can induce IBMIR.NAA has a protective effect on the hUC-MSCs in the infusion damage by inhibiting IBMIR, reducing the wastage of the blood components and enhancing the survival rate of the hUC-MSCs.

AIM:To investigate the influence of continuous subculturing of human umbilical cord mesenchymal stem cells (hUC-MSCs) on the mRNA expression of all 23 family members of NOD-like receptors (NLRs), and to search for the way of improving the subculture quality of hUC-MSCs and increasing the quantity and safety in the experimental and clinical application .METHODS:Neonatal umbilical cord was collected to isolate and purify the hUC-MSCs with the colla-genase II digestion and adherence screening methods .These cells were continuously subcultured .The hUC-MSCs at pas-sage 3 and passage 28 were identified by flow cytometry and induced differentiation .The mRNA expression of NLRs in the passage 3 and passage 28 hUC-MSCs was detected by RT-qPCR.RESULTS: The cell phenotypes of both passage 3 and passage 28 hUC-MSCs were CD29 +/CD44 +/CD105 +/CD31 -/CD34 -/CD40 -/CD45 -/CD106 -/HLA-DR-, and both of the cells were induced into osteoblasts and adipocytes , which were conformed to the criteria of International Society for Cellular Therapy to define MSCs .All the NLR family members were expressed in passage 3 hUC-MSCs.NOD1, NLRC4, NLRC5, NLRP1, NLRP3, NLRP10, NAIP, NLRX1 and APAF1 at mRNA levels were highly expressed , and the rest were lowly expressed.When hUC-MSCs were subcultured to passage 28, NLRP10 mRNA was increased, NLRC5 mRNA and NLRX1 mRNA were hardly changed , and all of the rest members were decreased .The difference of NLRP1 mRNA expres-sion between passage 3 and passage 28 hUC-MSCs was observed with statistical significance (P<0.05).CONCLU-SION:The effects of subculturing on the expression of NLR family in hUC-MSCs are pleiotropic .It requires further investi-gation to confirm whether these effects are related to the proliferation , differentiation and immunomodulation of MSCs .

Objective To explore the correlation of the radius and respiratory variation of inferior vena cava(IVC)with central venous pressure(CVP)for rapid evaluation of blood volume with ultrasound in elderly patients with septic shock.Methods The radius of IVC was measured using bedside ultrasound,respiration variation index(RVI)was calculated as following:RVI =(maximum radius-minimum radius)/maximum radius × 100％ and central venous pressure(CVP)was also recorded in 28 elderly patients with septic shock before and during 2 h and 6 h fluid recovery.Radius and RVI of IVC were compared between 28 shock patients and 22 healthy volunteers as control.Correlation of radius and RVI of IVC with CVP were analyzed.The thresholds of radius and RVI of IVC to estimate CVP 8 mmHg were determined by Receiver Operator Characteristic Curve (ROC)curves.Results The maximum and minimum radius[(1.23±0.28)cm and(0.48±0.18)cm]in the elderly patients with septic shock were smaller than in control group[(1.95±0.14)cm and (1.73±0.13)cm].RVI in the elderly patients with septic shock were larger than in control group [(55.88±11.18)％ vs.(11.23± 1.82)％].The maximum and minimum radius were positively(r=0.668 and 0.863,both P＜0.01)and RVI negatively(r=-0.848,P＜0.01)with CVP.The thresholds of maximum radius,minimum radius and RVI of IVC to estimate CVP 8 mmHg were 1.56cm(sensitivity 85.2％,specificity 86.3％),1.13 cm(sensitivity 96.3％,specificity 94.1％)and 30％(sensitivity 88.2％,specificity 96.3％),respectively.Conclusions Using ultrasound to measure radius of IVC and calculate RVI might estimate CVP to certain degree.It might be an option for physicians to rapidly estimate blood volume in the elderly patients with septic shock.

Objective To explore the correlation between radius and respiratory variation of inferior vena cava(IVC)and hemodynamic monitoring values of pulse-indicated continuous cardiac output(PiCCO)in septic shock pigs.Methods A total of 8 pigs were used to establish animal model of septic shock by intravenous infusing LPS(100 μg/kg),and fluid resuscitation was followed with normal saline.Ultrasound was used to measure the maximum radius(IVCmax)and minimum radius(IVCmin)of IVC,and respiration variation index(RVI)was calculated at basic status,septic shock,1 hour and 6 hours after fluid resuscitation,respectively.Respiratory variation index of IVC were calculated as:RVI =(IVCmax-IVCmin)/ IVCmax × 100％.Hemodynamic monitoring values,including ITBV,GEDV,SVV and CI of PiCCO,were recorded at the same time.Radius and RVI of IVC and PiCCO values between before and after fluid resuscitation were compared by LSD-t test.Correlation between radius and RVI of IVC andhemodynamic monitoring values were calculated by Pearson correlation coefficient.Results Compared with the moment of septic shock,IVC IVCmin,GEDV,ITBV and CI at 1 after hour fluid resuscitation were larger(P ＜ 0.01)and SVV and IVCrvi were smaller(P ＜ 0.01).Compared with the moment of septic shock and 1 after hour fluid resuscitation,IVC[VCmin,GEDV,[TBV and CI at 6 hours after fluid resuscitation were larger(P ＜ 0.01)and SVV and IVCrvi were smaller(P ＜ 0.01).IVCmax correlated with SVV(P=0.024)and it failed to correlate with GEDV,ITBV and CI.IVCmin correlated with GEDV(P=0.003),ITBV(P =0.001),SVV(P =0.009)and CI(P =0.015),respectively.RVI was correlated withGEDV(P＜0.01),ITBV(P＜0.01),SVV(P=0.007)and CI(P＜0.001),respectively.Conclusions Radius and RVI of IVC was correlated with hemodynamic monitoring values of PiCCO.It can serve as a parameter to rapidly estimate the blood volume.