Objective To investigate N-terminal pro-B-type natriuretic peptide (NT-proBNP) cutoff value for the mortality in different age groups in critically ill patients.Methods A retrospective study was conducted.295 patients admitted to the intensive care unit (ICU) of 401st Hospital of PLA from January 2011 to October 2012 were divided into two groups according to age [group with age＜65 years old (n=105) and group with age≥ 65 years old (n =190)].The concentrations of serum NT-proBNP,hematocrit (HCT),procalcitonin (PCT),C-reactive protein (CRP),serum creatinine (SCr),estimated glomerular filtration rate (eGFR),acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and probability of survival (PS) were recorded within 24 hours.The primary outcome was ICU mortality.Receiver operator characteristic curve (ROC curve) was used to evaluate the value of NT-proBNP for predicting the mortality.Results ① There were no significant differences in the length of stay in ICU,mechanical ventilation rate,the mortality,the incidence of cardiovascular disease,digestive disease,neurologic disease,and the number of patients having received operation,HCT,PCT and CRP between the two groups (all P＞0.05).The percentage of the male,the APACHE Ⅱ score,the percentage of respiratory disease,and NT-proBNP in group with age ≥ 65 years old were higher than those of the group with age ＜ 65 years old [the percentage of the male:51.6％ vs.33.0％,x2=9.093,P=0.003; APACHE Ⅱ score:22.94 ±8.10 vs.19.44 ±8.51,Z=-3.259,P=0.001; the percentage of respiratory disease:29.47％ vs.17.14％,x2=5.472,P=0.024; NT-proBNP(ng/L):5 859.00(2 050.75,23 802.75) vs.2 882.00 (275.15,6 236.00),Z=-5.514,P=0.000]; PS,the percentage of patients having multiple injuries and other diseases and eGFR in group with age ≥65 years old were lower than those of the group with age ＜65 years old [PS:59.0 (31.5,79.0)％ vs.70.0 (40.0,84.0),Z=-3.431,P=0.001; the percentage of multiple injuries:0.53％ vs.17.14％,x2=30.987,P=0.000; the percentage of other disease:5.79％ vs.13.33％,x2=4.962,P=0.030; eGFR (ml·min-1· 1.73 m-2):81.07 (45.77,131.80) vs.95.54 (33.64,165.55),Z=-2.214,P=0.027].② The area under the ROC curve (AUC) [95％ confidence interval (95％ CI)] of NT-proBNP in patients with age＜65 years old was significantly higher than that of group with age≥65 years old and the entire group [0.825(0.738-0.892) vs.0.664 (0.592-0.731) and 0.725 (0.670-0.775),Z1 =-2.835,P1 =0.005; Z2=-1.995,P2=0.046].③ The sensitivity (76.]0％ vs.64.10％),specificity (82.35％ vs.67.12％),positive predictive value (90.0％ vs.75.8％),and negative predictive value (62.2％ vs.53.8％) with cutoff value of NT-proBNP (2 882 ng/L) in group with age ＜65 years old were significantly higher than those with NT-proBNP cutoff value (6 062 ng/L) in group with age ≥ 65 years old.Conclusion NT-proBNP cutoff value in different age groups for the prediction of mortahty in the critically ill patients maybe more objective and accurate.

ABSTRACT:Objective To investigate the mechanism and induction of T type calcium channel on neural stem cells after brain injury .Methods Adult mice brain injury model was established and divided into control ,sham operation ,surgery ,and surgery+mebefradil groups .Neural stem cells were separated from the subventricular zone (SVZ) and identified .Moreover ,we analyzed the results using MTT and neural stem cell sphere counting after adding different doses of mibefradil in culture medium ,respectively .Then we calculated the half maximal inhibitory concentration (IC50) of mibefradil on neural stem cells .Finally ,we analyzed the expression of Cav3 .2 in SVZ and the protein expressions of Cav3 .2 ,Cyclin A and caspase‐3 in neural stem cells by Western blot .Results In vivo , neural stem cell proliferation was increased in surgery group compared with that in control and sham‐operation groups .The proliferation of neural stem cells in surgery + mibefradil groups was significantly decreased compared with that in surgery group after mibefradil‐induced inhibition of T type calcium channel protein . In vitro , the formation of neural stem cell sphere was significantly inhibited after adding mibefradil .The cell growth ratio was significantly decreased when the concentration of mibefradil was above 5μmol/L .A values in 5 μmol/L ,10 μmol/L and 20μmol/L groups were significantly lower than those in control group (P<0 .05) .IC50 was 8 .93μmol/L .The protein expressions of Cyclin A and Cav3 .2 were inhibited while that of caspase‐3 was increased after mibefradil treatment .Conclusion Neural stem cell proliferation was enhanced by activating T type calcium channel after brain injury .

Objective To evaluate the effects of midazolam pretreatment on inflammatory responses and cell apoptosis during intestinal ischemia-reperfusion (I/R) in mice.Methods Thirty healthy male Kunming mice,weighing 18-22 g,were equally and randomly divided into 3 groups using a random number table:sham operation group (S group),I/R group,and midazolam pretreatment group (M group).Intestinal I/R was produced by occlusion of the superior mesenteric artery for 20 min followed by reperfusion.In group M,midazolam 1 mg/kg was injected intraperitoneally,and intestinal I/R was produced 30 min later.At 24 h of reperfusion,the mice were sacrificed,and intestinal tissues were removed for microscopic examination and for determination of the expression of interleukin-6 (IL-6),tumor necrosis factor-alpha (TNF-α) and caspase-3.Intestinal damage was assessed and scored according to Chiu.Results Compared with group S,Chiu's scores were significantly increased,and the expression of IL-6,TNF-α and caspase-3 was significantly up-regulated in I/R and M groups (P＜0.05).Compared with group I/R,Chiu's scores were significantly decreased,and the expression of IL-6,TNF-α and caspase-3 was significantly down-regulated in group M (P＜0.05).Conclusion Midazolam pretreatment can reduce intestinal I/R injury,and the mechanism is related to inhibition of inflammatory responses and cell apoptosis in mice.

Objective:To observe the effects of acupuncture at Houxi(SI3)and Huantiao(GB30)on the expression levels of nuclear factor kappa B(NF-κB),inducible nitric oxide synthase(iNOS),and nitric oxide(NO)of NF-κB inflammatory signaling pathway in L5 spinal nerve root of lumbar disc herniation(LDH)model rats and explore the mechanism of acupuncture in LDH treatment.Methods:Forty specific-pathogen-free healthy male Sprague-Dawley rats were randomly divided into a sham operation group,a model group,acupuncture group 1,and acupuncture group 2,with 10 rats in each group.The non-compression nucleus protrusion model was made by puncturing L4-L5 spinous process space and injecting autologous nucleus suspension.Acupuncture at bilateral Shenshu(BL23),Dachangshu(BL25),and Weizhong(BL40)was carried out in acupuncture group 1,and acupuncture at bilateral Houxi(SI3)and Huantiao(GB30)in acupuncture group 2.All rats were treated with balanced reinforcing and reducing needling manipulations,and the needles were retained for 30 min/time with one episode of needling manipulation every 10 min,once a day,14 times in total.The threshold value of paw withdrawal pain was measured by a thermal stimulation pain instrument;the serum NF-κB,iNOS,and NO levels were measured by enzyme-linked immunosorbent assay.The pathomorphological changes of spinal nerve roots were observed by hematoxylin-eosin(HE)staining;quantitative reverse transcription polymerase chain reaction was used to detect iNOS mRNA expression in spinal nerve roots;the NF-κB and iNOS protein expression in spinal nerve roots was detected by the immunofluorescence method.Results:Compared with the sham operation group,the threshold of paw withdrawal pain in the model group was decreased,and the expression levels of serum NF-κB,iNOS,and NO were increased;HE staining showed many degenerated and dissolved Schwann cells in spinal nerve roots with vacuolar changes;meanwhile,the expression levels of NF-κB and iNOS proteins,and the iNOS mRNA in spinal nerve roots were increased.Compared with the model group,the paw withdrawal pain thresholds in acupuncture group 1 and acupuncture group 2 were increased,and the increase in acupuncture group 2 was greater(P<0.05);the expression levels of serum NF-κB,iNOS,and NO in acupuncture group 1 and acupuncture group 2 were decreased,especially in acupuncture group 2(P<0.01);the vacuolar changes of spinal nerve roots,and the degeneration and lysis of Schwann cells in acupuncture group 1 and acupuncture group 2 were decreased,which were more obvious in acupuncture group 2;the NF-κB and iNOS protein expression and the iNOS mRNA expression levels in spinal nerve roots of acupuncture group 1 and acupuncture group 2 were decreased,especially in acupuncture group 2(P<0.01).Conclusion:Acupuncture at Houxi(SI3)and Huantiao(GB30)can improve the morphology of spinal nerve roots,inhibit the NF-κB and iNOS protein expression levels in spinal nerve roots and the serum NO level,and relieve the pain caused by inflammation of spinal nerve roots,which may be one of the mechanisms of acupuncture in LDH treatment.

It has need to separate red blood cells (RBC) from marrow graft in ABO group unmatched BMT and auto-BMT with purging tumor cells, the separating effect of methylcellulose was observed. The mixture of 0.5% methylcellulose and bone marrow was laid up in an open transfusion system, and then sedimentation of RBC was performed in the transfusion tube. The separating results of 18 marrow grafts showed that the recovery rates of mononuclear cells and CD34(+) cells were (83.8 +/- 55.2)% and (90.3 +/- 7.2)%, respectively. RBC residual rate was (4.3 +/- 1.5)%. The yield of CFU-GM was (60.8 +/- 22.4)/2 x 10(5) MNC, and there was no difference to [(69.8 +/- 23.4)/2 x 10(5) MNC] yielded from same marrow samples, separated by Ficoll-Hypaque separation. It is concluded that this method could be used for bone marrow transplantation.
Bone Marrow Transplantation ; methods ; Cell Separation ; methods ; Erythrocytes ; immunology ; Humans ; Methylcellulose ; pharmacology

Background and purpose:Perioperative anesthetic management is thought to be critical to the success of free flap breast reconstruction. The purpose of this study was to discuss intraoperative fluid, hemodynamic and temperature management in patients undergoing deep inferior epigastric perforator (DIEP) flap breast reconstruction.Methods:From Jun. 2011 to Dec. 2015, 126 patients underwent DIEP lfap breast reconstruction. Postoperative complications were reviewed. Intraoperative fluid infusion rate was analyzed. Mean arterial blood pressure (MAP) and core temperature were measured before induction (T0), after lfap elevation but before lfap transfer (T1), 15 min after flap revascularization (T2), and at the end of surgery (T3).Results:Nine patients developed flap compromised: 7 were salvaged and 2 failed. The mean intraoperative lfuid infusion rate was (5.44±1.66) (mL?kg-1)/h. MAP at T0, T1, T2 and T3 were (87.45±8.90), (74.19±8.63), (74.60±8.71) and (79.62±7.88) mmHg, respectively. Core temperature at T0, T1, T2 and T3 were (36.69±0.14), (36.36±0.18), (36.27±0.14) and (36.21±0.15)℃, respectively. Conclusion:Standard practice focusing on intraoperative lfuid management, hemodynamic adjustment and temperature control in microsurgical reconstruction of the breast should be established to further improve free lfap outcome.

OBJECTIVETo construct TIMP-1 siRNA eukaryotic expression vectors and evaluate their effect on TIMP-1 mRNA expression in hepatic stellate cells.

METHODSThe combinant lone DNA with cutting sites of BamH I and Xho I enzyme according to the sequences of 447-465, 552-540 TIMP-1 of rats and nonspecific sequence were selected and cloned to pGEM-T vector and sub-cloned to pRNAT-U6.2. They were then identified by double enzyme digestion analysis and DNA sequencing. Three plasmids were transfected into T6 separately through an oligofectamine package. TIMP-1 mRNA expression was evaluated by RT-PCR.

RESULTSTargeting sequences of TIMP-1 siRNA eukaryotic expression vectors were correct. TIMP-1 mRNA expression was significantly reduced by transfecting them into the T6.

CONCLUSIONWe successfully constructed two TIMP-1 siRNA eukaryotic expression vectors and the transfected cells can significantly suppress the TIMP-1 expression.

Animals ; Cell Line ; Gene Silencing ; Genetic Vectors ; Hepatic Stellate Cells ; Plasmids ; RNA, Small Interfering ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; Transfection

Objective To study the effect of relatively closed environment dispensing mode on insoluble particles in pharmaceuticals.Methods The numbers of insoluble particles in conventional manual dispensing mode and relatively closed environment dispensing mode in lass Ⅰ and class Ⅲ environment were detected, effect of different dispensing modes on the number of insoluble particles in pharmaceuticals was compared.Results When adopting manual dispensing, the numbers of 1-10 μm insoluble particulates in different groups in class Ⅰ environment were less than those in class Ⅲ environment respectively, differences were all statistically significant (all P＜0.05). When adopting a new mode of dispensing in relatively closed environment, there was no significant difference in the numbers of 1-15 μm insoluble particulates in different groups between class Ⅰ and class Ⅲ environment respectively (all P＞0.05). The numbers of 1-20 μm and 1-15 μm insoluble particles in different groups were less than those in manual dispensing when the new mode of dispensing were adopted in class Ⅲ and class Ⅰenvironment respectively, differences were all statistically significant (all P＜0.05). The numbers of 1-15 μm insoluble particles in different groups in manual dispensing in class Ⅰ environment were more than those in class Ⅲ environment respectively, difference were all statistically significant (all P＜0.05).Conclusion The relatively closed environment dispensing mode can effectively reduce environmental particulates entering intravenous infusion system during dispensing process.

OBJECTIVETo explore the clinical value of Lens culinaris agglutinin-reactive alpha-fetoprotein in the differentiation diagnosis between benign and malignant liver diseases, as well as the early warning of hepatocellular carcinoma.

METHODSAlpha-fetoprotein variants from 300 patients with liver diseases were isolated with micro-spin column equipped lens culinaris agglutinin (LCA). The AFP and AFP-L3 were detected by the electrochemical luminescence (ECL) method, and the proportions of AFP-L3 were calculated.

RESULTSThe positive rates of AFP-L3 of HCC patients and chronic liver disease patients were 95% and 64% respectively, there were significant difference in two groups (chi2 = 134.72, P < 0.01), the HCC incidence rates of AFP-L3 positive and negative chronic liver disease patients showed significant difference (chi2 = 80.158, P < 0.01). there were no correlations between the proportion of AFP-L3 and AFP consistency(r = 0.046, P > 0.05).

CONCLUSIONSThe detection of AFP-L3 by micro-spin column assay show great clinical value in the differentiation diagnosis of benign and malignant liver diseases, as well as the early warning of hepatocellular carcinoma.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; analysis ; Carcinoma, Hepatocellular ; diagnosis ; Child ; Diagnosis, Differential ; Female ; Humans ; Liver Diseases ; diagnosis ; Liver Neoplasms ; diagnosis ; Male ; Middle Aged ; Plant Lectins ; chemistry ; Young Adult ; alpha-Fetoproteins ; analysis

OBJECTIVETo evaluate the usefulness of new microspincolumn method for the measurement of a1pha-fetoprotein variant AFP-L3 in differentiation of benign and malignant liver disease and the warming for liver cancer.

METHODSAFP-L3 was isolated by using microspincolumn coupled with lens culinaris agglutinin (LCA), AFP and AFP-L3 were determined with chemiluminescent immunoassay, the proportion of AFP-L3 levels AFP-L3(%) were calculated, and the relationship between the elevated AFP-L3(%) levels and benign and malignant liver disease was analyzed.

RESULTSThe levels of AFP-L3(%) in serum of patients with hepatocellular carcinoma was significantly higher than those in the patients with other liver diseases (P < 0.001). Taking AFP-L3(%) >or= 10% as the diagnostic criteria, the sensitivity for diagnosis of liver cancer was 90.9%.

CONCLUSIONDetection of AFP-L3 seemed to be of clinical value in diagnosis and differential diagnosis of hepatocellular carcinoma; it may be especially important for identifying patients with hepatocellular carcinoma whose a1pha-fetoprotein level is low.

Adult ; Aged ; Carcinoma, Hepatocellular ; blood ; diagnosis ; Diagnosis, Differential ; Female ; Hepatitis, Chronic ; blood ; diagnosis ; Humans ; Immunoassay ; methods ; Liver Cirrhosis ; blood ; diagnosis ; Liver Neoplasms ; blood ; diagnosis ; Luminescent Measurements ; methods ; Male ; Middle Aged ; Sensitivity and Specificity ; Young Adult ; alpha-Fetoproteins ; analysis