Plasma level of KPTT,factor Ⅷ coagulant activity,factor Ⅷ-related antigen,fib-rinogen,antithrombin Ⅲ activity,antithrombin Ⅲ antigen,tissue-plasminogen activator activity (t-PA:C),plasminogen activator inhibitor activity(PAI:C),PAI:C/t-PA:C,plasmin activity (PLm:C),D-dimer and plasma platelet α granule membrane protein were ditermined in 26 patients treated by long-term peritoneal dialysis(PD group),17 patients with chronic renal failure under-going conservative therapy(CRF group),36 patients on maintenance hemodialysis(HD group) and 20 age & sex-matched normal controls(NC group).The results showed:1.PD group had hypercoagulable state than NC group.The prothrombotic state in PD patients was most severe in contrast with CRF patients and HD patients.2.When PD group was compared with NC group,PLm:C showed no significant differences,but they had secondary fibrinolysis following enhanced coagulation.Moreover,PLm:C in PD group was significantly depressed as compared with in CRF group.3.the activated platelets in the PD patients were significantly elevated as compared with in the normal controls.

Objective To investigate the causes of acute renal failure(ARF)after orthtopic liver transplantation(OLT)in patients of acute liver failure(ALF)and the effects of systemic therapy based on continuous renal replacement(CRRT).Methods Clinical data of 412 patients who underwent liver transplantations between January 2001 and June 2006 were analyzed retrospectively (all the cases were followed up to June 2007).According to UNOS grading scale,54 patients were of acute liver failure(UNOS 1 and 2A).Posttransplant ARF developing in 17 cases underwent a systemic therapy based on CRRT as well as anti-rejection,anti-infection and nutrition support.The perioperative courses,complications,causes of death and follow up results were analyzed.Results There were no severe complications during CRRT.Perioperative mortality was 5.4%and 58.8%in patients without ARF and those with ARF respectively.the rate of complications was 35.1%vs 100%.1 year survival rate Was 89.2% vs 41.2%.3 year survival rate was 81.1% vs 41.2%.Condusions The effect of surgery mainly depends on the function of liver and other vital organs.The ALF recipients suffered from a high perioperative mortality,especially those with posttransplant ARL.The systemic therapy based on CRRT benefits patients with postoperative ARF.

Since there is a clinical need for the tissue-engineered vascular graft (TEVG), fabricating the vascular scaffold individually appears to be necessary. In this work, we have developed the traditional tubular scaffold and branch vascular scaffold utilizing low-temperature deposition manufacturing (LDM) technology. Then different tubular scaffolds were fabricated by changing the processing parameters, and the morphological properties of the scaffolds were assessed. The scaffolds reproduced the structure of 3D vascular model accurately. Wall thickness of the scaffold increased with the increase of velocity ratio (V(L)/V(s)) and nozzle temperature, and both the micropore size and wall roughness were positively correlated with the nozzle temperature. However, the porosity was barely affected by the nozzle temperature. This approach, fabricating vascular scaffold with special structure and appearance features via LDM technology, is potential for the individual fabrication of vascular scaffold.
Biocompatible Materials ; chemistry ; Blood Vessel Prosthesis ; Cold Temperature ; Computer-Aided Design ; Humans ; Lactic Acid ; chemistry ; Polyesters ; Polymers ; chemistry ; Tissue Engineering ; methods ; Tissue Scaffolds

OBJECTIVETo investigate if tumor necrosis factor-alpha (TNF-alpha) could induce phenotypic transformation or the associated cell function changes of human peritoneal mesothelial cells (HPMCs).

METHODSAll tests were performed on the human peritoneal mesothelial cell line (HMrSV5, kindly donated by Prof. Pierre RONCO). Morphological changes of HPMCs were observed by phase contrast microscopy. The expressions of cytokeratin 8, cytokeratin 18 and vimentin were detected by immunocytochemistry. mRNA expressions of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Production of MMP-9 and type I collagen was measured by enzyme-linked immunosorbent assays.

RESULTSTNF-alpha (1 - 10 ng/ml) induced morphological changes in about 50% - 60% of human peritoneal mesothelial cells (HPMCs). The original polygonal cobblestone monolayer was changed into elongated, spindle-shape characteristic of fibroblasts. The original strong positive expression of cytokeratin 8 and cytokeratin 18 was changed in all morphologically changed HPMCs to negative, but the expression of vimentin maintained positive. By 4-hour treatment with TNF-alpha (1 - 10 ng/ml), mRNA expression of MMP9 was significantly up-regulated, and mRNA expression of TIMP-1 and TIMP-2 were down-regulated. We also observed that the production of MMP-9 and type I collagen increased significantly after 24-hour treatment with TNF-alpha.

CONCLUSION24-hour treatment with TNF-alpha, produced a partial transformation of HPMCs into the fibroblast phenotype. TNF-alpha treatment of HPMCs up-regulated the synthesis of MMP-9 and type I collagen, which may facilitate peritoneal extracellular matrix remodeling and fibrogenesis.

Cell Line ; Cell Size ; drug effects ; Collagen Type I ; drug effects ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Immunohistochemistry ; Keratins ; analysis ; Matrix Metalloproteinase 9 ; genetics ; Peritoneum ; cytology ; drug effects ; metabolism ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinases ; genetics ; Tumor Necrosis Factor-alpha ; pharmacology ; Vimentin ; analysis

Objective To investigate the effects of different dialysates on expression of protein kinase C-δ (PKCδ) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKCδ specific inhibitor)group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKCδ mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were significantly higher than those at corresponding time points in blank control group , normal control group and fluid A+powder B+rottlerin group (P<0.05). Compared with normal control group, blank control group and fluid A+powder B+rottlerin group, the expression of PKCδ mRNA and protein of U937 cells in fluid A+powder B group were significantly increased (P<0.05). There was no significant difference in cell apoptosis rates and expression of PKCδ mRNA and protein between fluid A+fluid B group and blank control group, normal control group and fluid A+fluid B+rottlerin group (P＞0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKCδ. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing apoptosis of peripheral blood monouclear cells.

OBJECTIVE: To investigate if the DFNB59 gene contributes to the hearing loss of a Chinese pedigree with dominantly inherited auditory neuropathy (AN). METHOD: Nine members in four generations of the family were selected for this study. Genomic DNA was isolated from the peripheral leukocytes of the patients using the pure gene DNA isolation kits. Firstly, the subjects DNA fragment was PCR amplified using specific primers corresponding to exon 2 and 4 of the DFNB59 gene. Each fragment was purified and subsequently analyzed by direct sequencing in an applied biosystems 3730 automated DNA sequencer. The whole coding sequence of DFNB59 gene of one family patient were then PCR amplified and submitted for sequence analysis as described above. The resultant sequence data were compared with the standard sequence to identify deafness-associated mutations. RESULT: PCR amplifications were successfully conducted in all the subjects. We failed to detect the presence either of mutations T54I and R183W in the exon 2 and exon 4 that have been reported, or any other deafness-associated mutations in the whole DFNB59 gene, by sequence analysis. CONCLUSION The DFNB59 gene seems not contribute to the pathogenesis of this Chinese AN family, which suggesting new gene(s) involvement.
Asian Continental Ancestry Group ; genetics ; Base Sequence ; DNA Primers ; Female ; Humans ; Male ; Mutation ; Nerve Tissue Proteins ; genetics ; Pedigree ; Sequence Analysis, DNA ; Vestibulocochlear Nerve Diseases ; genetics

OBJECTIVE: To investigate if the OTOF gene contributes to the non-syndromic hearing loss of a Chinese pedigree with dominantly inherited auditory neuropathy (AN). METHOD: The subjects included were 9 live individuals in an autosomal dominant AN pedigree, 3 sporadic AN patients and 3 normal-hearing controls. Genomic DNA was isolated from the peripheral leukocytes of the subjects using the Pure gene DNA Isolation Kits. Firstly, the whole coding sequence of OTOF gene of one family patient were PCR amplified using specific primers. Each fragment was purified and subsequently analyzed by direct sequencing in an Applied Biosystems 3 730 automated DNA sequencer. The resultant sequence data were compared with the standard sequence to identify deafness-associated mutations. Other DNA samples were then screened for these mutations by PCR amplification and sequence analysis. RESULT: PCR amplifications were successfully conducted in all the subjects. Comparison of the resultant OTOF sequence in one family patient with the standard sequence identified 10 nucleotide variants which do not lead to amino acid change. These mutations were also detectable in other family individuals, 3 sporadic AN patients and 3 normal-hearing controls. CONCLUSION The OTOF does not seem to contribute to the pathogenesis of this Chinese AN family, which suggest new gene(s) involvement.
Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Chromosome Disorders ; ethnology ; genetics ; DNA Mutational Analysis ; Female ; Genetic Testing ; Hearing Loss ; genetics ; Humans ; Male ; Membrane Proteins ; genetics ; Mutation ; Pedigree ; Sequence Analysis ; Vestibulocochlear Nerve Diseases ; ethnology ; genetics

Objective To investigate the effects of Shenqi Tiaoshen Formula(SQTSF)for alleviating airway inflammation in rats with both chronic obstructive pulmonary disease(COPD)and lung-kidney qi deficiency syndrome and explore its therapeutic mechanism.Methods Forty-eight SD rats were randomly divided into control group,model group,low-,medium-,and high-dose SQTSF groups,and aminophylline(APL)group.In all but the control group,rat models of COPD with lung-kidney qi deficiency syndrome were established and treated with saline,SQTSF or APL via daily gavage as indicated(starting from day 30).The rats were observed for changes in body weight,grip strength,lung function,lung pathology,inflammatory cytokines in bronchoalveolar lavage fluid(BALF),oxidative stress levels,iron ion metabolism,cellular and mitochondrial ultrastructural changes in the lung tissue,and expressions of Nrf2/SLC7A11/GPX4 signaling pathway and ferroptosis-related proteins.Results The rats in the model group exhibited obvious symptoms of lung-kidney qi deficiency syndrome with significantly decreased body weight,grip strength,and lung function parameters.Examination of the lung tissue revealed showed significant inflammatory cell infiltration and emphysema with obvious bronchial,perivascular,and alveolar inflammation and alveolar destruction,significantly increased IL-1β,TNF-α,IL-6,and IL-13 levels in BALF,and elevated pulmonary oxidative stress levels and Fe2+and total iron ion concentrations.The rat models also showed characteristic ultrastructural changes of ferroptosis in the lung tissue cells under transmission electron microscope and significantly decreased Nrf2,GPX4,and SLC7A11 and increased ACSL4 expressions in the lung tissue.Treatment with SQTSF significantly improved these pathological changes in the rat models with a better effect than APL.Conclusion SQTSF can effectively improve airway inflammation and oxidative stress in COPD rats with lung-kidney qi deficiency possibly by inhibiting ferroptosis via regulating the Nrf2/SLC7A11/GPX4 signaling pathway.

Objective To investigate the effects of Shenqi Tiaoshen Formula(SQTSF)for alleviating airway inflammation in rats with both chronic obstructive pulmonary disease(COPD)and lung-kidney qi deficiency syndrome and explore its therapeutic mechanism.Methods Forty-eight SD rats were randomly divided into control group,model group,low-,medium-,and high-dose SQTSF groups,and aminophylline(APL)group.In all but the control group,rat models of COPD with lung-kidney qi deficiency syndrome were established and treated with saline,SQTSF or APL via daily gavage as indicated(starting from day 30).The rats were observed for changes in body weight,grip strength,lung function,lung pathology,inflammatory cytokines in bronchoalveolar lavage fluid(BALF),oxidative stress levels,iron ion metabolism,cellular and mitochondrial ultrastructural changes in the lung tissue,and expressions of Nrf2/SLC7A11/GPX4 signaling pathway and ferroptosis-related proteins.Results The rats in the model group exhibited obvious symptoms of lung-kidney qi deficiency syndrome with significantly decreased body weight,grip strength,and lung function parameters.Examination of the lung tissue revealed showed significant inflammatory cell infiltration and emphysema with obvious bronchial,perivascular,and alveolar inflammation and alveolar destruction,significantly increased IL-1β,TNF-α,IL-6,and IL-13 levels in BALF,and elevated pulmonary oxidative stress levels and Fe2+and total iron ion concentrations.The rat models also showed characteristic ultrastructural changes of ferroptosis in the lung tissue cells under transmission electron microscope and significantly decreased Nrf2,GPX4,and SLC7A11 and increased ACSL4 expressions in the lung tissue.Treatment with SQTSF significantly improved these pathological changes in the rat models with a better effect than APL.Conclusion SQTSF can effectively improve airway inflammation and oxidative stress in COPD rats with lung-kidney qi deficiency possibly by inhibiting ferroptosis via regulating the Nrf2/SLC7A11/GPX4 signaling pathway.

ObjectiveTo establish and evaluate a chronic obstructive pulmonary disease （COPD） model with lung-spleen qi deficiency. MethodA rat model mimicking COPD with lung-spleen qi deficiency was established by the combination of cigarette smoking and intratracheal instillation of lipopolysaccharide （LPS） along with gavage of Sennae Folium infusion. Forty male SPF-grade SD rats were randomly assigned to blank， model， and low- （L-FXY）， medium- （M-FXY）， and high-dose （H-FXY） Sennae Folium infusion groups. Other groups except the blank group were exposed to daily cigarette smoke， with LPS administrated via intratracheal instillation on the 1st and 14th days. On the 28th day of modeling， the L-FXY， M-FXY， and H-FXY groups were administrated with Sennae Folium infusion at 5， 10， and 20 g·kg^-1， respectively， and at 4 ℃ for three weeks. The modeling lasted for 49 days. The general conditions （body mass， food intake， fecal water content， and anal temperature） and behaviors （grip strength test and tail suspension test） of rats in different groups were examined. The lung function， lung histopathology， D-xylose， amylase， and gastrin levels in the serum， interleukin（IL）-1β and IL-6 levels in the alveolar lavage fluid， levels of T-lymphocyte subsets （CD4⁺， CD8⁺， and CD4⁺/CD8⁺） in the peripheral blood， and thymus and spleen indices were measured. ResultTwo rats died in the H-FXY group. Compared with the blank group， both the M-FXY and H-FXY groups exhibited reduced body mass and food intake （P<0.01） and increased fecal water content （P<0.01）. The anal temperature in the H-FXY group was lower than that in the blank group （P<0.01）. The grip strength decreased in the modeling groups compared with the blank group （P<0.01）， and the duration of immobility in the tail suspension test increased in the M-FXY and H-FXY groups （P<0.05， P<0.01）. Compared with the blank group， the modeling groups showed reduced 0.3 second forced expiratory volume （FEV_0.3）， FEV_0.3/forced vital capacity （FVC）（P<0.01）， thickening of bronchial walls， proliferation of goblet cells， and the presence of emphysematous changes. In terms of gastrointestinal function， the M-FXY and H-FXY groups had lower levels of D-xylose， gastrin， and α-amylase than the blank group （P<0.01）. Regarding the immune and inflammatory indices， the M-FXY and H-FXY groups showed lower thymus and spleen indices than the blank group （P<0.01）. Compared with the blank group， the modeling groups presented lowered CD4⁺ level （P<0.01） and CD4⁺/CD8⁺ ratio （P<0.05， P<0.01） in the peripheral blood and elevated levels of IL-1β and IL-6 in the alveolar lavage fluid （P<0.01） than the blank group. ConclusionA model of COPD with lung-spleen Qi deficiency was established through the combination of daily cigarette smoke， intratracheal instillation with LPS， and gavage of Sennae Folium infusion. The comprehensive evaluation results suggested medium-dose （10 g·kg^-1） Sennae Folium infusion for gavage during the modeling of COPD with lung-spleen Qi deficiency.