Objective To investigate clinical significance of plasma D-dimer (D-D),fibrinogen (FIB) and fibrin/fibrinogen degradation product (FDP) in patients with chronic obstructive pulmonary disease (COPD).Methods The level of plasma D-D,FIB and FDP in 150 patients with COPD and 80 healthy persons were detected,and compared.Results The level of plasma D-D,FIB and FDP in COPD patients were significantly higher than those in healthy persons[(2.16 ± 0.61) mg/L vs.(0.55 ± 0.04) mg/L,(5.88 ± 1.52) g/L vs.(3.12 ± 0.35) g/L,(7.18 ± 1.63) mg/L vs.(3.62 ± 1.55) mg/L],there were significant differences (P ＜ 0.01).Conclusion Monitoring the level of plasma D-D,FIB and FDP in COPD patients can provide reliable basis in hypercoagulable state and primary and secondary hyperfibrinolysis.

With the foreign students into the clinical practice stage,how to improve the teaching quality is very important for teachers. Sufficient preparations before practice; adopting PIO teaching methods; training the exact thinking of the clinical medicine and examing students'medical case records in detail are important factors to the successful teaching.

Objective To investigate the effects of rapamycin on renal tissue and function of rats with protein overload nephropathy and to explore the protective mechanism of losartan. Methods Experimental rat models with protein overload nephropathy, induced by intraperiotoneal injection of BSA (2. 0 g/d)into female Wistar rats, were divided into three groups: control group, rapamycin group(injected intraperitoneally with rapamycin) and losartan group(injected intraperitoneally with rapamycin and given orally with losartan). At different time points, the quantity of 24-hour urine protein and renal function were measured, and the morphologic changes of renal tissues were evaluated by HE staining and electron microscope. Results Both at day 7 and day 14, rats received BSA developed intense proteinuria. At day 7, compared with control group, 24-hour proteinuria increased markedly in rapamyein group (P<0.05). Nevertheless,proteinufia was notably alleviated in losartan group (P<0.05). At day 14, 24-hour-urine protein of rapamycin group was also significantly higher than that of the losartan group (P<0.05), but therewas no significant difference between control group and losartan group (P>0.05). Proteinuria and intratubular albumin cast formation were alleviated notably in losartan group. The fusion of focal podocytes in rapamycin group was obvious in comparison with control group. Conclusions Rapamycin can agrravate proteinuria in rats with protein overload nephropathy through changing the barrier of glomerular filtration by damaging podocytes. Furthermore, losartan can alleviate severe proteinuria induced by rapamycin.

Objective To investigate the effect of tumor necrosis factor ot (TNF-α) on monocyte-renal tubular epithelial cell adhesion and to explore its associated mechanism. Methods Human renal proximal tubular epithelial cell line (HK2 cells)and monocyte cell line (U937)were used to perform cell co-culture. TNF-α (10 μg/L)was used to stimulate HK2 cells and then U937 was put into the HK2 monolayers. Three locational parts of hyaluronan around HK2 cells were extracted by different ways and the HA expression was detected by enzyme-linked binding protein assay. Monocyte adhesion was detected by florescence recording assay. Flow cytometry was applied to assess intercellular adhesion molecule-1 (ICAM-1) expression in HK2 cells. Results Stimulation of HK2 cells with TNF-α significantly increased U937 binding to HK2 cell monolayer [(2.25±0.05) folds vs control, P<0.01]. Incubation of TNF-α (10 μg/L) with HK2 cells for 24 hours did not induce the change of total HA expression surrounding the HK2 cells, whereas HA redistribution happened with conditioned medium fraction (CM-HA) increased [(1.17±0.16) fold vs control, P<0.05], the pericellular fraction (trypsin extract fraction, TE-HA) decreased (83%±11% vs control, P<0.05) and the remaining cells-associated HA (cell associated fraction, CA) did not change significantly. In addition, pre-treatment of HK2 cells with TNF-α significantly increased the ICAM-1 expression [(1.85±0.22) folds vs control, P<0.01] and ICAM-1-dependent monocytes binding. Removal of HA from the surface of confluent monolayers of HK2 cells by hyaluronidase before addition of U937 cells increased monocyte binding[ (1.35±0.06) folds vs control, P<0.05]. In contrast, the presence of either sICAM-1 (400 μg/L) or anti-CDI8 antibody (10 mg/L) led to a significant decrease in ICAM-1-dependent monocyte binding (78%±7%, P<0.05; 75%±8%, P<0.01 vs those before adding sICAM-1 or anti-CD18 antibody, respectively). Conclusion HA redistribution induced by TNF-α may facilitate ICAM-1-dependent monocyte-epithelium binding, and pericellular HA plays a protective role in monocyte-driven tissue inflammation.

Objective:To investigate the effects of antimalarial artemether on the proliferation of human lung adenocarcinoma cell line A549 in vitro and provide experimental data for the treatment of lung cancer with artemether.Methods:MTT assay was used to observe the inhibitory effects of artemether on the proliferation of A549 cells.Cell growth curve was draw according to the cell counts.The population doubling time was obtained in logarithmic growth phase.Cell cycle detection was observed by flow cytometry.H-E staining and transmission electron microscopy were used to observe the altered morphology of apoptotic cells. Results:Artemether has a significantly inhibitory effect on the proliferation of A549 cells in a dose-dependent manner in vitro,and the IC50 was 1.34 mg/L.The population doubling time in logarithmic growth phase in the artemether treatment group was(20.7?0.5) h compared to(32.2?0.3) h in the control group.The difference between two groups was statistically significant(P

0.05).The optimal combination was PCR-CTPP for MDR1 C3435T and PCR-SSP for G2677T/A.Conclusions PCR-CTPP and PCR-SSP are simple,accurate,rapid and economical methods for detection of SNP of MDR1 C3435T and G2677T/A,and can be applied in clinical research.

Objective To investigate the risk factors of chylous leakage after pancreatioduodenectomy so as to find effective measures to prevent this complication.Methods A retrospective analysis was conducted on 230 patients who underwent pancreatioduodenectomy at the First Affiliated Hospital of Zhejiang University from Jun.2012 to Jun.2014.Patients with chylous leakage were identified and a 1 ∶ 2 patients in the study and the control groups were selected.The parameters for matching included tumor volume,vascular invasion,and extent of lymph node dissection.A logistic analysis was performed to identify independent risk factors of chylous leakage.Results 15 (6.5％) patients developed chylous leakage after pancreatioduodenectomy.The average hospital stay after surgery of the study group was 20.8 days,compared to 13.5 days in the control-group (P =0.004).In the study group,chylous leakage rate increased in patients with 14th and 16th group of lymph nodes dissection (80％ vs 36.7％,P =0.006).Logistic analysis showed that 14th and 16th lymph nodes dissection was an independent risk factor of chylous leakage after pancreatioduodenectomy (P ＜ 0.05,OR =6.909,95％ CI 1.593 ～ 29.958).Conclusions Chylous leakage prolonged hospitalization after pancreatioduodenectomy.Dissection of the 14th and 16th lymph node groups was an independent risk factor of chylous leakage after pancreatioduodenectomy.Careful ligation of the gastrocolic vein near the lymphatic trunk and dissection of 14th and 16th group of lymph nodes were effective interventions to reduce postoperative chylous leakage.

Objective To investigate the effect of albumin on expression of NLRP3 inflammasome and its downstream cytokines IL-1β and IL-18 in tubular epithelial cells.Methods Thirty mesangioproliferative glomerulonephritis (MsPGN) patients with different levels of proteinuria were selected, and their renal biopsy samples were stained by PAS and Masson to observe tubular epithelial cells injury and inflammatory cells infiltration.NLRP3, caspase-1, IL-1β and IL-18, as well as different inflammatory cells, were detected by immunohistostaining.In vitro, Western blotting and real-time PCR were employed to detect NLRP3, caspase-1, IL-1β and IL-18 protein and mRNA in HK-2 cells stimulated by bovine serum albumin (BSA) (20 g/L).Results In MsPGN patients with high levels of proteinuria, there were obvious renal tubular epithelial cell injury and inflammatory cells infiltration (all P ＜ 0.05), and the expressions of NLRP3, caspase-1, IL-1β and IL-18 were up-regulated compared to patients with low levels of proteinuria (all P ＜ 0.05).Furthermore, IL-1β and IL-18expressions were positively correlated with the degree of proteinuria (r=0.836, P ＜ 0.05;r=0.901, P ＜0.05).NLRP3, caspase-1, IL-1β and IL-18 protein and mRNA were significantly increased in HK-2cells stimulated by BSA compared to the control group (all P ＜ 0.05).Conclusions Albumin is able to induce NLRP3 inflammasome activation in tubular epithelial cells, which may be the mechanism of tubulointerstitial injury and inflammation caused by proteinuria.

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Objective To investigate the effects of nephroblastoma over-expressed protein (CCN3) on the formation of extracellular matrix (ECM) induced by transforming growth factor-β1 (TGF-β1) in human mesangial cells (HMCs) and its underlying signal transduction mechanism related with microRNA-29(miRNA-29).Methods HMCs were pretreated with different doses of exogenous CCN3 (5 μg/L,50 μg/L and 500 μg/L) or transfected with pcDNA3.1(+)-CCN3 before exposed to TGF-β1(2 μg/L),to observe the expression of fibronectin (FN),type I collagen (COL I) and miRNA-29a,b and c.The mimics or inhibitor of the miRNA-29a were transfected into HMCs to analyze whether miRNA-29a affect CCN3.The expressions of FN mRNA,COL I mRNA and miRNA-29 family were detected by real time PCR.The protein expressions of FN and COL I were detected by Western blotting and cell immunofluorescence.Results (1) Compared with the normal control group,the expressions of FN and COL I were up-regulated in TGF-β1 group,while the expressions of miRNA-29a,b,c were down-regulated in TGF-β1 group (all P ＜ 0.05).(2) Compared with the TGF-β1 group,the expressions of FN and COL I were decreased when pretreated with the different doses of exogenous of CCN3 or transfected with pcDNA3.1(+)-CCN3 (all P ＜ 0.05).Meanwhile,the expression of miRNA-29a was significantly increased when pretreated with 50 μg/L and 500 μg/L CCN3 or transfected with pcDNA3.1(+)-CCN3 (all P ＜ 0.05);whereas miRNA-29b and c had no statistical difference (all P ＞ 0.05).(3) Compared with TGF-β1+CCN3 group,the expressions of FN and COL I were decreased in CCN3+TGF-β1+miRNA-29a mimics group (all P ＜ 0.05),whereas the expressions of FN and COL I in CCN3+TGF-β1+miRNA-29a inhibitors group were increased (all P ＜ 0.05).Conclusions CCN3 reduces the TGF-β1-induced production of ECM by the up-regulation of miRNA-29a.