Objective To evaluate age-related changes of early plasma renal injury markers in rats with sepsis, which can serve as evidence for the early detection of acute kidney injury (AKI). Methods Twenty-one male Sprague Dawley rats were divided into 3 groups based on their age: young group(12 weeks, n=7), adult group (12 months, n=7), and old group (D-galactose-induced 12 months, 500 mg/(kg.d), for 6 weeks, n=7). All rats were received cecal ligation and puncture (CLP) procedure to induce sepsis. Plasma samples of each rat were collected 0.5 h before CLP, and 0.5 h, 4.5 h, 8.5 h after CLP. For each sample, plasma level of kidney injury markers, including plasma creatinine (Cr), blood urea nitrogen (BUN), neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), interleukin-6 (IL-6), Cystatin C (CysC), insulin-like growth factor-binding protein 7 (IGFBP-7), tissue inhibitor of metalloproteinases-2 (TIMP-2) and [TIMP-2]*[IGFBP-7] were quantified. We adopted the criteria for AKI: plasma creatinine elevation =1.5 times of the baseline level. Within-group longitudinal changes and between-group changes were evaluated. Results Compared with samples collected at 0.5 h before CLP, plasma Cr levels were significantly increased at 0.5 h, 4.5 h and 8.5 h after CLP in all three groups (P<0.05), while plasma IL-6 [(121.1±14.9) pg/ml vs. (98.24±13.37) pg/ml, P=0.009] and BUN [(5.43±0.73) mmol/L vs. (5.06±0.82) mmol/L, P=0.041] significantly decreased at 4.5 h after CLP in adult rats, Only the young rats suffered AKI. In between-group comparison, plasma level of IL-6 [(98.24±13.37) pg/ml vs. (116.86±19.75) pg/ml, P=0.046], TIMP-2 [(4042.56±781.68) pg/ml vs. (5662.57±1527.8) pg/ml, P=0.013], [TIMP-2]*[IGFBP-7] [(0.112±0.03) (ng/ml)2/1000 vs. (0.17±0.052) (ng/ml)2/1000, P=0.021] significantly increased at 4.5 h after CLP and BUN [(5.34±0.43) mmol/L vs. (6.29±0.51) mmol/L, P=0.017] at 8.5 h after CLP in the young group, compared with the adult rats. There was no significant difference in the plasma levels of other markers when compared with 0.5 h before CLP and also no significant between-group difference (P>0.05). Conclusion Only the young rats suffered earlier and more serious sepsis-induced AKI. Plasma level of Cr may be useful for early detection of sepsis. Age may not be a risk factor for early AKI in sepsis. Plasma TIMP-2, [TIMP-2]*[IGFBP-7] are potential markers for the detection of early AKI in sepsis in both young and adult animals.

Objective To identify the potential risk factors affecting mortality rate of ALl in severe trauma population. Method It was a retrospective cohort study treating trauma as a single cause for emergency depart-ment (ED)) and emergency intensive care unit (EICU) admissions. Eighteen potential risk factors affecting the mortality of ALI were examined by univariate and multivariate logistic analyses in these severe trauma patients. Re-sults There were 343 severe trauma patients with post-traumatic ALI admitted to ED and EICU the Second Affili-ated Hospital Medical College,Zhejiang University,during the study period. The five risk factors that affected the mortality with unadjusted odd ratios (ORs) and 95% confidence intervals (CIs) were (1) APACHE Ⅱ score, (2)duration of trauma, (3) age, (4) aspiration of gastric contents, and (5) DIC. Specific risk factors also affected different patients subpepulations at different degrees. Conclusions Factors of APACHE Ⅱ score and aspiration of gastric contents that can predict the mortality of ALl may exist in the early stage of trauma. Duration of trauma and DIC that greatly affect the short- and long-term development of ALI deserve special attention. Elderly patients (aged beyond 65 years) are the independent risk factor for the secondary sepsis and deterioration of pulmonary function. Patients with these risk factors need aggressive supportive care as early as possible in order to prevent fur-ther aggravation.

Professional assessment in Chinese Higher Education has made great progress in three stages: the sporadic practice, trial and promotion. The authors present several comments on the characteristics and the professional assessment standards of clinical pharmacy in China, and focus on the scientific system of professional assessment.

OBJECTIVEThe aim of this paper was to evaluate the treatment outcome of multimodal treatment for 196 patients with locoregional recurrent esophageal cancer after curative treatment and to determine the prognostic factors of recurrence.

METHODSOne hundred and ninety six patients with locoregional recurrent esophageal cancer curatively treated in our hospital were included in this study. Kaplan-Meier method was used to analyze the survival rate. Log rank test was used to evaluate the difference between the groups. Multivariate survival analysis was conducted using a Cox proportional hazard regression model with a backward stepwise procedure.

RESULTSThe overall 1-, 2- and 3-year survival rates were 29.8%, 5.9% and 4.0%, respectively, with a median survival time of 8.0 months. The univariate analysis showed that ECOG PS, the interval between initial treatment and recurrence, the regimens of initial treatment and retreatment were independent prognostic factors. The multivariate analysis showed that the regimens of initial treatment and retreatment were independent prognostic factors. Retreatment methods significantly influenced the survival. The median survival time of chemoradiotherapy, radiation therapy alone, chemotherapy alone, EGFR-TKI and best supportive care were 13.0, 7.0, 6.0, 4.0 and 3.0 months, respectively (P = 0.000).

CONCLUSIONSThe prognosis of patients with locoregional recurrent esophageal cancer after curative treatment is poor. The main prognostic factors are the regimens of initial treatment and retreatment. Multimodal treatment including radiotherapy and chemotherapy may improve the long-term survival of the patients.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Squamous Cell ; therapy ; Chemoradiotherapy ; Combined Modality Therapy ; Esophageal Neoplasms ; therapy ; Esophagectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; therapy ; Proportional Hazards Models ; Radiotherapy, High-Energy ; Retrospective Studies ; Survival Rate

OBJECTIVETo evaluate the hemocompatibility of glutaraldehyde (GA)-tanned bovine pericardium additionally treated by sodium bisulfite (SOB) solution.

METHODSThe hemocompatibility of GA-tanned bovine pericardium treated by SOB solution is evaluated by using dynamic clotting time test, blood platelet adhension test, D-dimeride determination, and complement activation test. The GA-tanned bovine pericardium was used as control.

RESULTSThe curve of absorbance-clotting time of two kinds of bovine pericardium was similar in dynamic clotting time test. There was no significant difference between SOB-treated and control groups in blood platelet adhension test. The D-dimeride contents of all bioprostheses were at normal level, and the D-dimeride content of GA-tanned bovine pericardium treated by SOB solution was significantly lower than that of control group (P < 0.05). In complement activation test, the level of complement C3a in SOB-treated group was significantly lower than that in control group (P < 0.05).

CONCLUSIONGA-tanned bovine pericardium treated by SOB solution meets the demands of cardiac interstitial implanted materials in hemocompatibility.

Animals ; Biocompatible Materials ; Bioprosthesis ; Blood Coagulation ; Cattle ; Complement C3a ; analysis ; Fibrin Fibrinogen Degradation Products ; metabolism ; Glutaral ; pharmacology ; Materials Testing ; Pericardium ; drug effects ; metabolism ; Platelet Adhesiveness ; Sulfites ; pharmacology

OBJECTIVETo investigate the possibility to enhance the proliferation of adipose-derived stem cells (ASCs) in a delayed fat flap in rabbits.

METHODSA delayed fat flap was formed in one side of inguinal region of a rabbit. 21 days after operation, the fat tissues at the delayed flaps and at the unoperated side were harvested and digested with 0.25% collagenase and sieved. The cell suspensions were centrifuged. The cells were obtained from tissue precipitate after centrifugation. The expression rates of the surface marker (CD29, CD44, CD14 and CD45) were measured by FCM and compared between the experimental and control groups.

RESULTSExpression rates of CD29 and CD44 were higher in the delayed fat flap (74.06% and 90.74%) than in the contralateral fat tissue (62.88% and 77.54%, P < 0.05), while those of CD14 and CD45 were lower in the delayed fat flap (57.66% and 4.84%) than in the contralateral fat tissue (72.10% and 75.82%, P < 0.05 and P < 0.01).

CONCLUSIONSTissue hypoxic ischemia such as fat tissue in a delayed fat flap can promote proliferation of ASCs. It indicates that tissue in the delayed flap may be transplanted with better survival rate. The ischemia pretreatment of fat tissue may become a new method for fat transplantation.

Adipose Tissue ; cytology ; transplantation ; Animals ; Cell Proliferation ; Cells, Cultured ; Graft Survival ; Postoperative Period ; Rabbits ; Stem Cells ; cytology ; Surgical Flaps

Embryonic hematopoiesis in mammals is characterized by successive temporal and spatial changes. Previous investigations indicate that in vitro differentiation of embryonic stem cells (ES cells) derived from 129 mice can mimic embryonic hematopoiesis to some extent. To investigate the in vitro hematopoietic differentiation capacity of ES cells derived from C57BL/6 mice, the authors initially established the murine ES cell line with standard identification methods employed. Next, two-step culture system was utilized for embryoid bodies formation and the appearance of different hematopoietic precursors was confirmed by CFC assay, cellular chemical staining as well as RT-PCR. The results demonstrated that the ES cell line MES-1 fulfilled the criteria of ES cell line and its progeny after in vitro differentiation included primitive and definitive erythrocyte precursors, mixed colony-forming cells and granulocyte/macrophage colony-forming cells. RT-PCR analysis revealed the molecular consistence of transcription factors and hematopoietic markers with cellular event. In conclusion, MES-1 established from C57BL/6 mice was able to differentiate in vitro to a variety of hematopoietic precursors, thus could partly recapitulate embryonic hematopoiesis.
Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; genetics ; Cell Line ; Colony-Forming Units Assay ; DNA-Binding Proteins ; genetics ; Embryo, Mammalian ; cytology ; Erythroblasts ; cytology ; metabolism ; Erythroid Precursor Cells ; cytology ; metabolism ; Erythroid-Specific DNA-Binding Factors ; Gene Expression ; Hematopoietic Stem Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism ; Time Factors ; Transcription Factors ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; genetics

BACKGROUNDVascular endothelial growth factor (VEGF) is well known as a hypoxia-induced protein. That it markedly increased expression of VEGF and improvement of rat motor function after spinal cord injury suggested that VEGF could play a neuroprotective role in ischaemic tolerance. This study investigated whether vascular endothelial growth factor has direct neuroprotective effects on rat spinal cord neurons.

METHODSWe employed primary cultures of embryonic rat spinal cord neurons, then administrated different concentrations of VEGF164 in the culture medium before hypoxia when the number of neurons was counted and the cell viability was detected by MTT. The neuronal apoptosis and expression of VEGF and its receptor genes were evaluated by terminal deoxynucleotidyl transferase mediated dUTP nick-end labelling (TUNEL) and immunohistochemistry. The VEGFR2/FLK-1 inhibitor, SU1498, was used to confirm whether the neuroprotective effect of VEGF was mediated through VEGFR2/Flk-1 receptors.

RESULTIn hypoxic conditions, the number and viability of neurons decreased progressively, while the number of TUNEL-positive cells increased along with the prolongation of hypoxic exposure. When the concentration of VEGF in cell culture medium reached 25 ng/ml, the cell viability increased 11% and neuronal apoptosis reduced to half, this effect was dose dependent and led to an approximately 25% increase in cell viability and about threefold decrease in TUNEL-positive cells at a maximally effective concentration of 100 ng/ml. In normal conditions, VEGF/Flk-1 but not VEGF/Flt-1 gene expressed at a low level: after hypoxia, the expression of VEGF/Flk-1, but not VEGF/Flt-1 was significantly increased. The protective effect of VEGF was blocked by the VEGFR2/Flk-1 receptor tyrosine kinase inhibitor, SU1498.

CONCLUSIONSVEGF has direct neuroprotective effects on rat spinal cord neurons, which may be mediated in vitro through VEGFR2/Flk-1 receptors.

Animals ; Cell Hypoxia ; Cells, Cultured ; Cinnamates ; pharmacology ; Female ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; drug effects ; Vascular Endothelial Growth Factor A ; pharmacology ; Vascular Endothelial Growth Factor Receptor-2 ; physiology

OBJECTIVETo investigate the effect of cytomegalovirus (CMV) infection on actin and microfilament in human embryo fibroblast cells (HF) and its relationship with CMV replication.

METHODSCell morphology was observed after the infection of CMV. Western-blot was used to measure the expression levels of beta-actin, G-actin and F-actin proteins. CMV immediately early antigen (CMV IE) in HF cells was analyzed by indirect immunofluorescence assay. Microfilament alteration was determined by cytoskeleton fluorescence probe.

RESULTCMV IE was demonstrated in more than 95% of HF cells after infection, which was primarily located in nucleus. The shape of HF cells changed from thin shuttle like to round and thick ball like, even escaping from wall after infection by CMV. Compared with control group, the expression of G-actin protein increased at 24 h of CMV infection (0.941 +/-0.061 compared with 0.714 +/-0.119, P <0.05), then decreased at 72 h, 96 h respectively(0.218 +/-.035, 0.230 +/-0.055 compared with 0.714 +/-0.119, P <0.05). The levels of F-actin in infected cells gradually decreased at 24 h, 72 h and 96 h compared with control HF cells (0.256 +/-0.021, 0.127 +/-0.032, 0.026 +/-0.008 compared with 0.373 +/-0.050, P<0.05). In infected HF cells, microfilaments were found ruptured, arranged turbulently. Cells fused and fluorescence density of microfilament markedly reduced.

CONCLUSIONCytomegalovirus can induce alteration of actins and microfilament, which may be associated with its infection, replication and reactivity in host cells.

Actin Cytoskeleton ; metabolism ; Actins ; biosynthesis ; genetics ; Antigens, Viral ; analysis ; Cells, Cultured ; Cytomegalovirus ; Cytoskeleton ; metabolism ; Embryo, Mammalian ; Fibroblasts ; metabolism ; ultrastructure ; virology ; Humans ; Immediate-Early Proteins ; analysis

OBJECTIVETo analyze characteristic CT enhancement patterns of noncalcified pulmonary tuberculomas and their pathological basis.

METHODFifty-six patients with noncalcified pulmonary tuberculomas underwent surgical resection of the tuberculomas. Enhanced CT images of these tuberculomas were reviewed and analyzed in relation to the histological findings.

RESULTSOf the 56 patients, 45 showed no enhancement in the tuberculomas, which were histologically characterized by central caseous necrosis and a poorly vascularized peripheral fibrotic zone. Eleven patients showed ring-like or eggshell enhancement, and the central low density region was histologically confirmed to be caused by caseous or liquefied necrosis, while the ring enhancement resulted pathologically from moderately or well vascularized peripheral fibrotic or granulomatous tissues.

CONCLUSIONSPulmonary tuberculomas consists mainly of caseous necrotic tissues characterized by no enhancement and ring or eggshell enhancement on dynamic contrast-enhanced CT.

Adult ; Calcinosis ; Contrast Media ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed ; Tuberculoma ; diagnostic imaging ; metabolism ; Tuberculosis, Pulmonary ; diagnostic imaging ; metabolism ; Young Adult