OBJECTIVETo investigate the effect of continuous veno-venous hemodiafiltration (CVVHDF) on endotoxin-induced acute lung injury (ALI) of piglets.

METHODSEighteen piglets were randomly divided into three groups: control group (n = 6); heparin group (n = 6) and CVVHDF treatment group (n = 6). All the animals were anesthetized by muscle injection of ketamine (30 mg/kg), then placed in supine position, received continuous intravenous infusion of ketamine with the rate of 10 mg/(kgxh). After placing a 4.5 cm (inner diameter) tracheal tube via tracheostoma, controlled mechanical ventilation was established using the assisted-controlled ventilation option of the NEWPORT 200. Respiratory rate at 30 breath/min; PIP at 10 cm H2O (1 cm H2O = 0.098 kPa); PEEP at 2 cm H2O and fraction of inspired oxygen at 0.3. A vein catheter was placed into right vena jugularis interna to administer a Ringer's solution. Initially, at a rate of 10 ml/kg, followed by a rate of 15 ml/kg when the mean arterial blood pressure was below 70 mm Hg (1 mm Hg = 0.133 kPa), the rate of 20 ml/kg was used when the mean arterial blood pressure was below 60 mm Hg. An 8Fr double-lumen catheter was inserted into left femoral vein and served as the pathway for CVVHDF. A Pulsiocath Pcco catheter was positioned into left femoral artery to monitor the circulatory parameters. All catheters were flushed with heparinized saline to prevent clotting. Then all the animals were given intravenous infusion of 150 microg/kg endotoxin within 30 minutes to induce ALI. When the oxygenation index < 300 and pulmonary compliance < 30% of the baseline, the animals of heparin group received heparin infusion to maintain blood active coagulation time (ACT) 180 - 250 s, the animals of treatment group received CVVHDF with the blood flow of 50 ml/min, replacement rate of 300 ml/h, dialysis rate of 600 ml/h and the ultrafiltrate rate of 350 ml/h for six hours, heparin infusion to keep blood ACT 180 - 250 s. The circulatory parameters: heart rate (HR), mean arterial blood pressure (MABP), central venous pressure (CVP), pulse contour cardiac output index (PCCI); systemic venous resistance index (SVRI), cardiac function index (CFI), external venous lung water index (EVLWI), left ventricular contractile index (dPmx); respiratory parameters: respiratory rate (RR), pulmonary compliance (Cdyn) were monitored; arterial blood gas analysis was performed and oxygenation index (PaO2/FiO2) was calculated. All the parameters were recorded at baseline (B), onset of ALI (A 0 h), two hours (A 2 h), four hours (A 4 h), six hours (A 6 h) after ALI.

RESULTSNo significant difference in circulatory parameters, respiratory parameters and blood gas analysis were found at B and A 0 h among the three groups. When the ALI occurred, PaO2/FiO2, Cdyn, MABP and PCCI of the three groups decreased; HR, RR, EVLWI, SVRI increased. After four hours of ALI, the RR, EVLWI, SVRI, CFI and dPmx of treatment group were improved, the differences were significant compared with the other two groups (P < 0.05). After six hours of ALI, the HR, PCCI, MABP, PaO2/FiO2 and Cdyn of treatment group were significantly improved, compared with control group and heparin group (P < 0.05). There were no significant differences in any of the parameters between control group and heparin group. The difference in CVP among three groups was not significant.

CONCLUSIONCVVHDF has a good effect on hemodynamics of the endotoxin-induced ALI of the piglets.

Acute Lung Injury ; etiology ; physiopathology ; therapy ; Animals ; Endotoxins ; adverse effects ; Hemodiafiltration ; Hemodynamics ; Swine

OBJECTIVETo observe the effects of angiotensin II type 1 receptor blocker valsartan in preventing hepatic fibrosis induced by dimethylnitrosamine in rats.

METHODSExcept rats in the control group, all were given intraperitoneal injections of 1% dimethylnitrosamine (DMN 1 ml/kg, two or three consecutive days/a week for 6 weeks). From the first day of the intraperitoneal injection, rats in treatment groups were given valsartan for 8 weeks by gastric gavage. Liver tissue and blood samples of all rats were examined at 56 days (8 weeks). AngII levels were determined by radioimmunoassay. Hepatic mRNA levels of Collagen type I (Col I) and tissue inhibitor of metalloproteinase1 (TIMP1) were evaluated by reverse-transcription polymerase chain reaction (RT-PCR).

RESULTSValsartan significantly attenuated the degree of liver fibrosis and decreased the hepatic AngII content compared with DMN treated rats (P<0.01). mRNA levels of Col I and TIMP1 were upregulated in DMN treated rats compared with normal rats. Valsartan downregulated the elevation of Col I and TIMP1 mRNA levels (P<0.01).

CONCLUSIONHepatic AngII content of the model group was increased, the local tissue RAS was activated in DMN induced liver fibrosis. Valsartan can retard the progression of hepatic fibrosis and may provide an effective new strategy for anti-liver fibrosis therapy.

Angiotensin II Type 1 Receptor Blockers ; pharmacology ; therapeutic use ; Animals ; Dimethylnitrosamine ; Female ; Liver Cirrhosis, Experimental ; chemically induced ; prevention & control ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Tetrazoles ; pharmacology ; therapeutic use ; Valine ; analogs & derivatives ; pharmacology ; therapeutic use ; Valsartan

BACKGROUNDFor patients undergoing off-pump coronary artery bypass grafting (OPCABG), it is important to establish a hemodynamic monitoring system to obtain powerful parameters for better intraoperative treatment. This study aimed to observe the clinical feasibility of arterial pressure-based cardiac output (APCO) for cardiac output (CO) monitoring and to evaluate the correlation between APCO and pulmonary artery catheter (PAC) for CO measurement for patients undergoing OPCABG intraoperatively.

METHODSFifty patients of American Society of Anaesthesiologists (ASA) classification II-III, undergoing elective OPCABG at Beijing Anzhen Hospital were randomly enrolled into this study. All patients were assigned to CO monitoring by PAC and APCO simultaneously. Patients with pacemaker, severe valvular heart disease, left ventricular ejection fraction (EF) < 40%, cardiac arrhythmias, peripheral vascular disease, application of intra-aortic balloon pump (IABP) and emergent diversion to cardiac pulmonary bypass were excluded. The radial artery waveform was analyzed to estimate the stroke volume (SV) and heart rate (HR) continuously. CO was calculated as SV ' HR; other derived parameters were cardiac index (CI), stroke volume index (SVI), systemic vascular resistance (SVR), and systemic vascular resistance index (SVRI). PAC was placed via right internal jugular vein and the correct position was confirmed by PAC waveforms. Continuous cardiac output (CCO), CI and other hemodynamic parameters were monitored at following 5 time points: immediate after anesthesia induction (baseline value), anastomosis of left internal mammary artery to left anterior descending artery (LAD), anastomosis of left circumflex (LCX), anastomosis of posterior descending artery (PDA) and immediate after sternal closure.

RESULTSIn the 50 patients, preoperative echocardiography measured left ventricular EF was (52.8 ± 11.5)%, and 35 patients (70%) showed regional wall motion abnormalities. The correlation coefficient of CO monitored by APCO and PAC were 0.70, 0.59, 0.78, 0.74 and 0.85 at each time point. The bias range of CI monitored from both APCO and PAC were (0.39 ± 0.06) L×min(-1)×m(-2), (0.48 ± 0.12) L×min(-1)×m(-2), (0.26 ± 0.06) L×min(-1)×m(-2), (0.27 ± 0.06) L×min(-1)×m(-2), (0.30 ± 0.05) L×min(-1)×m(-2) at each time point. The results of SVR by two hemodynamic monitoring techniques had good correlation during OPCABG. The variation trends of SVR were opposite comparing with the results of CO. SVR collected from PAC obtained the highest value of (1220.0 ± 254.0) dyn×s×cm(-5) at PDA anastomosis, but the highest value obtained from APCO was (1206.0 ± 226.5) dyn×s×cm(-5) in LCX anastomosis.

CONCLUSIONSAPCO is feasible in hemodynamic monitoring for patients undergoing OPCABG. The results of hemodynamic monitoring derived from APCO and PAC are closely correlated. Its characterizations of timely, accurate and continuous display of hemodynamic parameters are also obviously demonstrated in the present study.

Aged ; Arterial Pressure ; physiology ; Cardiac Output ; physiology ; Catheterization, Swan-Ganz ; methods ; Coronary Artery Bypass ; methods ; Female ; Hemodynamics ; Humans ; Male ; Middle Aged ; Monitoring, Intraoperative ; methods

OBJECTIVESince continuous blood purification (CBP) has the effects of eliminating inflammatory mediators and improving organs function, CBP had been applied to treat non-renal diseases for nearly 10 years, but few studies have been conducted in children with sepsis and multiorgan dysfunction syndrome (MODS), especially in China. The present study aimed to evaluate the clinical effect of CBP in treatment of children with severe sepsis and MODS.

METHODSTwenty-two children with severe sepsis and MODS admitted to our PICU from Aug. 2003 to Aug. 2005 were treated with continuous veno-venous hemodialysis filtration. Their heart rate, arterial blood pressure, doses of vasoactive agents, spontaneous respiratory rate, PO2/FiO2 and prognosis were investigated.

RESULTSCatheterization and CBP were carried out in all the 22 children. Continuous vein-vein hemodialysis filtration (CVVHDF) and pre-dilution were chosen. The duration of CBP was (64.4 +/- 34.5) h. All the children had tachycardia before CBP and the heart rate fell gradually to 45 +/- 13 bpm 4 h after CBP. Blood pressure (BP) was stable in 7 children without shock during CBP. Ten children with early shock could maintain normal BP during CBP, but the doses of vasoactive agents were tapered 1 to 5 h after beginning of CBP and use of these agents was discontinued at 2 to 8 h. BP was elevated by (25.2 +/- 10.7) mmHg (1 mmHg = 0.133 kPa) in 5 refractorily shocked children 4 h after CBP and returned to normal level 8 h later. The doses of the vasoactive drugs were reduced at 2 to 8 h and ended 4 to 16 h later, which was longer than that of children with early stage shock. The accelerated spontaneous respiratory rate was slowed down by 7 +/- 4 per minute 4 h later, PO2/FiO2 rose from (177.7 +/- 53.1) mmHg before CBP to (341.0 +/- 60.2) mmHg 4 h after CBP in children with respiratory failure and reached the normal value (5.3 +/- 2.1) h later. FiO2 declined to less than 50%. Pediatric critical illness score was 62.2 +/- 7.4 on admission and elevated to (86.6 +/- 9.0) 24 h later, which was a significant elevation as compared to that of children with sepsis who were not treated with CBP seen between Aug. 2001 and July 2003. The survival rate was 72.7% after CBP and the effective rate of the treatment was 90.9%, but was 36% in children who were not treated with CVVHDF.

CONCLUSIONCBP can effectively improve the vital organ's function of children with sepsis and MODS and raise their survival rate. Replacement fluid of modified Ports formula was useful for stability of serum potassium and sodium, but resulted in elevation of serum glucose, calcium, and osmolarity. The application of CBP in children with sepsis can lead to slight drop of blood pressure at the beginning and to bleeding during CBP.

Adolescent ; Blood Pressure ; Child ; Child, Preschool ; Female ; Heart Rate ; Hemofiltration ; methods ; Humans ; Infant ; Infant, Newborn ; Intensive Care Units, Pediatric ; Male ; Multiple Organ Failure ; etiology ; mortality ; physiopathology ; therapy ; Prognosis ; Sepsis ; complications ; mortality ; physiopathology ; therapy ; Severity of Illness Index ; Survival Rate ; Treatment Outcome

BACKGROUNDA satisfactory animal model of breast cancer metastasizing to bone is unavailable. In this study, we used human breast cancer stem-like cells and human bone to build a novel "human-source" model of human breast cancer skeletal metastasis.

METHODSHuman breast cancer stem-like cells, the CD44+/CD24-/lower subpopulation, was separated and cultured. Before injection with the stem-like cells, mice were implanted with human bone in the right or left dorsal flanks. Animals in Groups A, B, and C were injected with 1 x 10(5), 1 x 10(6) human breast cancer stem-like cells, and 1 x 10(6) parental MDA-MB-231 cells, respectively. A positive control group (D) without implantation of human bone was also injected with 1 x 10(6) MDA-MB-231 cells. Immunohistochemistry was performed for determination of CD34, CD105, smooth muscle antibody, CD44, CD24, cytokine, CXC chemokine receptor-4 (CXCR4), and osteopontin (OPN). mRNA levels of CD44, CD24, CXCR4, and OPN in bone metastasis tissues were analyzed by real-time quantitative polymerase chain reaction (PCR).

RESULTSOur results demonstrated that cells in implanted human bones of group B, which received 1 x 10(6) cancer stem-like cells, stained strongly positive for CD44, CXCR4, and OPN, whereas those of other groups showed no or minimum staining. Moreover, group B had the highest incidence of human bone metastasis (77.8%, P = 0.0230) and no accompaniment of other tissue metastasis. The real-time PCR showed an increase of CD44, CXCR4, and OPN mRNA in metastatic bone tissues in group B compared with those of groups C and D, however the expression of CD24 mRNA in group B were the lowest.

CONCLUSIONSIn the novel "human source" model of breast cancer, breast cancer stem-like cells demonstrated a higher human bone-seeking ability. Its mechanism might be related to the higher expressions of CD44, CXCR4, and OPN, and the lower expression of CD24 in breast cancer stem-like cells.

Animals ; Bone Neoplasms ; secondary ; Breast Neoplasms ; pathology ; CD24 Antigen ; analysis ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Hepatocyte Growth Factor ; pharmacology ; Humans ; Hyaluronan Receptors ; analysis ; Immunohistochemistry ; Mice ; Neoplastic Stem Cells ; pathology ; Osteopontin ; analysis ; Phenotype ; Receptors, CXCR4 ; analysis

OBJECTIVETo explore the diagnosis and differential diagnosis of refractory cytopenia of children (RCC) according to WHO classification, and discuss the relationship between the cytology reviewed by hematologists and histology reviewed by pathologists.

METHODSWe selected 50 non-severe aplastic anemia cases from 2007 - 2010 in our hospital and collected clinical data. Experienced hematologists and pathologists evaluated bone marrow biopsy and smear respectively.

RESULTSOf 50 cases, 23 were male and 27 female (M:F = 1:1.17), the median age at diagnosis was 9 years (ranged from 3 to 14 years). 5 patients had disagreement of diagnosis between hematologists and pathologists. In 3 cases hematologists diagnosed as aplastic anemia (AA) and pathologists as RCC, 2 cases vice versa. The final diagnoses of 50 patients reached consensus between hematologists and pathologists were AA 16 cases, RCC 34 cases including 8 refractory cytopenias with multilineage dysplasia (RCMD) cases. All 16 cases AA showed severe hypocellularity. Only 4 cases (25.00%) RCC showed severe hypocellularity, 19 cases (73.08%) RCC showed mild hypocellularity and 3 cases (11.54%) RCC were normal hypocellularity.

CONCLUSIONOur results suggests that RCC was not rare in China. The main feature of RCC was dysplasia because of absence of increased blast. RCC was easily confused with AA. The main points of differential were present dysplastic changes of megakaryocyte best appreciated by the hematologists and morphologists and abnormal location of hematopoietic easily observed by pathologists. Overall, cytology and histology were complementary in the investigation of RCC and AA, because of sometimes one might give information that not be given from the other.

Adolescent ; Anemia, Aplastic ; diagnosis ; pathology ; Bone Marrow ; pathology ; Bone Marrow Examination ; Child ; Child, Preschool ; Diagnosis, Differential ; Female ; Humans ; Male ; Myelodysplastic Syndromes ; diagnosis ; pathology ; Pancytopenia ; diagnosis ; pathology ; Retrospective Studies

OBJECTIVETo evaluate the safety and efficacy of intracoronary autologous bone marrow mononuclear cells (BM-MNCs) transplantation in patients with dilated cardiomyopathy (DCM).

METHODSOn top of standard therapy, DCM patients received BM-MNCs transplantation (n = 71) or saline injection (n = 187). The baseline clinical characteristics of two groups were comparable. Data on echocardiography, Holter, six-minute-walk test, cardiac SPECT and annual hospital days were obtained in all patients at baseline, 1, 3, 6, 12 and 24 months after transplantation.

RESULTSSix-minute-walk distance was significantly longer at one month [(345 +/- 76) m vs. (286 +/- 104) m, P < 0.05] and thereafter (all P < 0.05) in BM-MNCs group compared with saline group. Left ventri ocular ejection fraction (LVEF) at one month in BM-MNCs group was significantly higher compared with saline group [(41.5 +/- 9.4)% vs. (37.3 +/- 6.6)%, P < 0.05] and with pre-transplantation value [(41.5 +/- 9.4)% vs. (32.4 +/- 8.5)%, P < 0.05] while LVEF was similar at 24 months after transplantation between the two groups [(43.6 +/- 6.3)% vs. (43.2 +/- 6.0)%, P > 0.05]. Three months after transplantation, the number of ischemic segments of BM-MNCs group was significantly reduced compared with that of saline group (2.0 +/- 1.0 vs. 3.1 +/- 1.4, P < 0.05) and with baseline (2.0 +/- 1.0 vs. 3.1 +/- 1.2, P < 0.05) while the number of necrotic segments were similar in both groups during the follow-up. There were no significant difference in survival between two groups during 2 years follow-up (95.4% vs. 94.9%, P > 0.05) but the annual hospitalization days of BM-MNCs group was significantly lower than that of saline group [(23.6 +/- 13.4) d vs. (33.0 +/- 14.0) d, P > 0.05].

CONCLUSIONSIntracoronary transplantation of autologous BM-MNCs was safe and could increase LVEF and the six-minute-walk distance and reduce hospitalization days for patients with dilated cardiomyopathy.

Adolescent ; Adult ; Aged ; Bone Marrow Transplantation ; methods ; Cardiomyopathy, Dilated ; therapy ; Female ; Follow-Up Studies ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Middle Aged ; Transplantation, Autologous ; Treatment Outcome ; Young Adult

Objective To study the effect and mechanism of berberine (BBR) on the lung metastasis of mouse breast cancer via epithelial-mesenchymal transition (EMT). Methods CCK-8 and Transwell migration assays were utilized to investigate the proliferation and migration properties of breast cancer 4T1 cells after BBR treatment.Mouse 4T1-Luc cells were injected into mice under the fourth mammary fat pad, and the mice were then randomly divided into the control and BBR groups.The mice in the BBR group received daily intraperitoneal injections of BBR working solution and those in the control group were continuously intraperitoneally injected with the same volume of the solvent used to dissolve BBR powder.Tumor metastasis in the lungs of living mice was detected by using an in vivo imaging system.After 42 days of administration, lung metastasis was measured via microscopy and HE staining.Western blot analysis was used to examine the effects of BBR on the expression of EMT-related proteins (Vimentin and Snail) as well as the activation of the Akt and ERK signaling pathways. Results BBR significantly promoted 4T1 cell migration (P < 0.05).In vivo experiments showed that the number of lung metastases in the BBR group had significantly increased compared with that in control group (P < 0.05) as observed under microcopy and histological staining.Compared with the control group, BBR upregulated the expression levels of Vimentin and Snail as well as the phosphorylated levels of p-Akt and p-ERK (P < 0.05). Conclusion BBR may promote EMT and lung metastasis of breast cancer 4T1 cells by activating the expression of proteins in the p-Akt and p-ERK pathways.

OBJECTIVESTo explore the dynamic changes and interactions between MMP-2 and TIMP-2 during experimental liver fibrosis.

METHODSWistar rats were randomly allocated into a normal group and a model group. To induce liver fibrosis, rats were injected intraperitoneally with dimethylnitrosamine (DMN) three consecutive times in the first week, then two consecutive times per week, totally for 6 weeks. In the normal control group, rats were injected with saline by the same method as the model group. Animals were sacrificed 1, 4, 10, 17, 28, 42, 56 days after starting DMN injections. Conventional histological examinations of the livers were performed with hematoxylin and eosin and Masson staining. The fibrosis was classified into 0 to 4 stages. Hydroxyproline content was determined after liver tissues were hydrolyzed in HCl at 160 degree C for 2 hrs and then measured with spectrometry at 560 nm wavelength. mRNA levels of MMP-2 and TIMP-2 were determined by semi-quantitive RT-PCR. Gelatinase activity of MMP-2 was examined by zymography using gelatin substrate.

RESULTSIn the model group the hepatic MMP-2 mRNA expression started to increase 10 days after DMN administration and remained at a much higher level than in the normal group throughout the study period, while TIMP-2 mRNA expression started to be lower than in the normal group 17 days after DMN administration and reached the lowest level on the 28th day. Then it rapidly rebounded and remained higher than that in the normal group from the 42nd day to the end of the study period. TIMP-2/MMP-2 began to be lower by several days than that of the normal group after DMN administration through the remaining study period. Zymography showed that the enzymatic activities of both latent MMP-2 and active MMP-2 were increased during the process of liver fibrosis.

CONCLUSIONIn liver fibrosis, MMP-2 expression increases, while TIMP-2 expression relatively decreases. The enzymatic activities of MMP-2 increase as the liver fibrosis develops.

Animals ; Female ; Liver Cirrhosis, Experimental ; enzymology ; metabolism ; Male ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-2 ; biosynthesis ; genetics ; metabolism

OBJECTIVETo study the pulmonary pathology in patients died of fatal human influenza A(H1N1) infection.

METHODSEight cases of fatal human influenza A (H1N1) infection, including 2 autopsy cases and 6 paramortem needle puncture biopsies, were enrolled into the study. Histologic examination, immunohistochemitry, flow cytometry and Western blotting were carried out.

RESULTSThe major pathologic changes included necrotizing bronchiolitis with surrounding inflammation, diffuse alveolar damage and pulmonary hemorrhage. Influenza viral antigen expression was detected in the lung tissue by Western blotting. Immunohistochemical study demonstrated the presence of nuclear protein and hemagglutinin virus antigens in parts of trachea, bronchial epithelium and glands, alveolar epithelium, macrophages and endothelium. Flow cytometry showed that the apoptotic rate of type II pneumocytes (32.15%, 78.15%) was significantly higher than that of the controls (1.93%, 3.77%).

CONCLUSIONNecrotizing bronchiolitis, diffuse alveolar damage and pulmonary hemorrhage followed by pulmonary fibrosis in late stage are the major pathologic changes in fatal human influenza A (H1N1) infection.

Adolescent ; Adult ; Aged ; Alveolar Epithelial Cells ; pathology ; Antigens, Viral ; metabolism ; Apoptosis ; Autopsy ; Biopsy, Needle ; Bronchiolitis, Viral ; pathology ; Child ; Child, Preschool ; Female ; Hemagglutinin Glycoproteins, Influenza Virus ; metabolism ; Humans ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza, Human ; metabolism ; mortality ; pathology ; virology ; Lung ; immunology ; metabolism ; pathology ; Male ; Middle Aged ; Nuclear Proteins ; metabolism ; Pulmonary Alveoli ; pathology ; Pulmonary Fibrosis ; pathology ; Young Adult