Objective To investigate the effects of continuous venovenous hemodiafiltration (CVVHDF) on plasma lipopolysaccharide (LPS),interleukin(IL) 1β,IL-4,high mobility group box 1 (HMGB-1) in multiple organ dysfunction syndrome (MODS) dogs,to explore the therapeutic mechnism of CVVHDF.Methods Dogs were subjected to hemorrhagic shock plus resuscitation and endotoxemia to establish MODS model,then they were randomly divided into 2 groups:CVVHDF group (n=6) and MODS group (n=6).After endotoxin injection completed,the CVVHDF group received CVVHDF treatment for 24 h; MODS group did not receive CVVHDF treatment.The LPS,IL-1β,IL-4,HMGB-1 levels in plasma and ultrafiltrate were measured at different time points(before operation,before intravenous infusion of endotoxin,0 h,3 h,6 h,9 h,12 h,18 h,24 h after intravenous infusion completion of endotoxin),then the sieving coefficient (SC) was calculated.Results Compared with MODS group,the LPS,IL-1β,IL-4,HMGB-1 levels in plasma were significantly decreased in the CVVHDF group at 9 h,12 h,18 h,24 h (P ＜ 0.05).Some concentration of IL-1β,IL-4 and HMGB-1 was detected in the ultrafiltrate,the SC for IL-1β,IL-4 and HMGB-1 was 0.60±0.12,0.83±0.11,0.70±0.09 respectively,and LPS could not be detected in the ultrafiltrate,its SC was 0.After treatment,the damage level of lung and renal had been significantly reduced in CVVHDF group.Conclusion CVVHDF can effectively reduce the levels of pro-inflammatory mediator and anti-inflammatory cytokine related to MODS,cut off the vicious circle of cytokine network and reduce the damage level of organism in the treatment of MODS.

Objective To investigate the expression of Survivin,receptor for advanced glycation endproduct(RAGE) and high mobility group protein B1 (HMGB1) gene in human breast cancer and their clinical significance.Methods The expression of Survivin,RAGE and HMGB1 gene was detected by Real-time PCR technology and fluorescence in situ hybridization (FISH) technology in the following tissue samples:50 early breast cancers,50 advanced breast cancers,and the corresponding adjacent normal mammary tissues.The relationship of their expression and several factors such as differentiation degree,invasion,lymph node metastasis and TNM stage of cancer was explored.Results The positive expression rate of Survivin,RAGE and HMGB1 gene was 62％,73％ and 79％ detected by Real-time PCR technology,78％,69％ and 72％ detected by FISH technology in breast cancer tissues.The overexpression of these genes was positively correlated with TNM stage and lymph node metastasis.The expression of Survivin gene was positively correlated with the expression of RAGE and HMGB1.Conclusion Overexpression of Survivin,RAGE and HMGB1 gene is of great significance in early diagnosis and prognosis of human breast cancer.

Ohjective To investigate the expression of Survivin gene in human breast cancer and its clinical significance.MethodsReal-time PCR technology and fluorescence in situ hybridization (FISH) technology were employed to detect the expression of Survivin gene in the following tissue samples:50 breast cancers in stage Ⅰ and Ⅱ,50 breast cancers in stage Ⅲ and Ⅳ,and the corresponding adjacent normal mammary tissues.The relationship of Survivin gene expression and several factors such as differentiation degree,invasion,lymph node metastasis,TNM stage was explored.ResultsThe positive expression rate of Survivin gene was 62％ and 78％ respectively detected by real-time PCR and FISH in breast cancer tissues.The overexpression of Survivin gene was positively correlated with TNM stage and lymph node metastasis.ConclusionsOverexpression of Survivin gene can be used as one of the parameters in early diagnosis and prognosis of human breast cancer.Selective gene therapy based on these gene expressions may be useful.

Objective Plasma phospholipid metabolism analysis was employed to investigate plasma metabolic profile of acute renal graft rejection in order to find potential disease biomarkers and reveal its pathophysiological changes.Methods Selected 33 patients which did renal transplant in our hospital during 2009 to 2010,named preoperative group.14 patients were diagnosed as acute graft rejection,and 19 patients were non-acute graft rejection.Then use ultra performance liquid chromatography (UPLC) coupled with quadrupole time of flight mass spectrometry (qTOF-MS) to establish the metabolic fingerprint in those patients.The date was preprocessed with software Markerlynx,and analyzed with partial least squares discriminant analysis (PLS-DA),and data from conventional laboratory techniques were used for assessing renal function.Results In the plasma of renal transplant patients,seventeen biomarkers were discovered:creatinine,four sphingomyelins (SM),nine phosphatidylcholines (PC),three lysophosphatidylcholine (LPC).While the creatinine was increased,the others were reduced obviously.Conclutions UPLC-qTOF-MS based metabonomics can analyze the plasma phospholipid metabolism in patients with renal transplantation,reveal the law of phospholipids metabolic changes in the process of acute rejection.The levels of plasma phosphatidylcholine's esterlysis and fatty acids' β-oxidation are increased.The immunosuppressive therapy can reduce the activity of phospholipase,inhibit the level of plasma phosphatidylcholine's esterlysis and fatty acids' β-oxidation,which restraines the development of acute rejection after renal transplantation.Therefore it may be a promising new technology in diagnosing acute renal graft rejection after renal transplantation.

To establish the online two-dimensional liquid chromatography by using double gradient liquid chromatography system and UV detector, in order to simultaneously determine the content of paeoniflorin, paenol, amygdaloside and cinnamic acid. A pump of the two-dimensional liquid chromatography was adopted as the one-dimensional separation pump. C18 (3.0 mm x 150 mm, 3 microm) was used as the analytical column, with acetonitrile as the organic phase and 0.08% phosphoric acid + 0.08% triethylamine as the aqueous phase for gradient elution at the flow rate of 0.5 mL x min(-1). Another pump of the two-dimensional liquid chromatography was adopted as the two-dimensional separation pump. PAII C18 was used as the analytical column, with acetonitrile as the organic phase and 20 mmol, pH 3.0 monopotassium phosphate as the aqueous phase for gradient elution at the flow rate of 0.8 mL x min(-1). The detection wavelengths were set at 218, 230, 275 nm by using wavelength time-switching program. The linearity range of paeoniflorin, amygdaloside, paeonol and cinnamic acid were 5.55-222 (r = 0.999 7), 6.6-264 (r = 0.999 8), 3.3-132 (r = 0.999 5) and 0.315-12.6 mg x L(-1) (r = 0.999 7), respectively. The average recoveries of the four components were between 96.12% and 103.9%. The experiment proved that this method was so rapid and accurate in determination results that it could be used for evaluating drug quality.
Capsules ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Online Systems ; Time Factors

This study aims to investigate the virological impact of the stalk region and cysteine (C) in neuraminidase (NA) of influenza A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1) viruses. The NA of A/ Anhui/1/05 (H5N1), defined as AH N1, lacked 20 amino acids (including C, defined as s20) as compared with NA of A/Ohio/07/2009 (H1N1) (defined as 09N1). We deleted s20 of 09N1 to construct 09N1-s20, and inserted s20 into AH N1 to construct AH N1+s20. To investigate the impact of C on the biological function of NA, we deleted C in 09N1 to construct 09N1-C and inserted C into AH N1 to construct AH N1-C. The pseudo-type viral particle (pp) system was used to evaluate the impact of these mutants on virology. The combination of 09N1-C and 09H1 (defined as 09H1::09N1-C) showed an infectivity 8 times that of the wild type 09H1::09N1, while the infectivity of the combination of AH N1+C and AH H5 (defined as AH H5::AH N1+C) was much lower than that of the wild type AH H5::AH N1. The infectivity of the combination of 09N1-s20 and 09H1 (defined as 09H1::09N1-s20) was 4 times that of the wild type 09H1::09N1; the infectivity of the combination of AH N1+s20 and AH H5 (defined as AH H5:: AH N1+s20) was 1/7 that of the wild type AH H5::AH N1. The co-existence of 09N1-C and AH H5 displayed 6 times the infectivity of AH H5::09N1, while the infectivity of 09H1::AH N1+C was very low. Multimer analysis showed that in the wild type 09N1, the forms of NA were dimer > tetramer > monomer; the major component of NA in 09N1-C was monomer; in 09N1-s20, the forms of NA were monomer > dimer. AH N1 was mainly composed of monomer; in AH N1+s20, the forms of NA were dimer > monomer > tetramer; in AH N1+C, the forms of NA were dimer > tetramer. Deletion of C or s20 from 09N1 did not change the expression of NA. The study suggested that deletion of C from the stalk region of NA in A/Ohio/07/2009 (H1N1) increases infectivity. Insertion of C into NA's stalk region of A/ Anhui/1/05 (H5N1) significantly decreases infectivity. Cysteine deletion in the stalk region is important for the infectivity of A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1). It may interfere with the infectivity via changes in NA polymerization.
Amino Acid Motifs ; Humans ; Influenza A Virus, H1N1 Subtype ; chemistry ; enzymology ; genetics ; pathogenicity ; Influenza A Virus, H5N1 Subtype ; chemistry ; enzymology ; genetics ; pathogenicity ; Influenza, Human ; virology ; Neuraminidase ; chemistry ; genetics ; metabolism ; Viral Proteins ; chemistry ; genetics ; metabolism ; Virulence

Objective To evaluate the safety and feasibility of minimally invasive percutaneous nephro-lithotomy (mPCNL) with U100plus laser for the treatment of renal calculi. Methods From October 2006 to December 2008 ,mPCNL was performed on 133 patients suffering from renal calculi by using Wolf 8/9. 8 rig-id ureteroscope and U10Oplus laser. Results mPCNL was completed in all the 133 cases. Residual calculi were found in 7 cases after operation and use medical drags to treat. The most residual calculi were removed after 1 month and a few stones being survived. 5 cases with residual calculi were treated by ESWL. The total stone clearance was 91.0%. The operation time was 38 -65 min(mean 46 min). Nephrostomy tube was kept for a mean of 1 -2 d. The mean postoperational hospital stay was 2 -4 d. Among the patients, 133 were followed up for 1-16 months (mean 8. 3 months) , during which no recurrent renal stones were found by B ultrasonngraphy or X ray. Conclusion By using mPCNL with U100plus laser, patients with renal calculi can be treated safely and effectively.

Objective To observe the anti-proliferation effects of rapamycin and paclitaxel of different hu-man prostate cancer cells in vitro. Methods The methods of MTr and flow cytometry were respectively ap-plied to observe the effect of rapamycin, paclitaxel and rapamycin+paclitaxel on proliferation and apoptosis of different prostate cancer cell lines (LNCaP-C4, LNCaP-C4-2, PC-3). Results When the concentration of rapamycin was 0.01 μmol/L, the impressive effect showed a remarkable difference in contrast to the con-trol. While in group LNCaP-C4-2 and PC-3, the goal concentration of rapamycin was 0.001μmol/L. When the concentration of paclitaxel was 0. 2 ng/mL, the impressive effect showed a remarkable difference in con-trast to the control. In group rapamycin (10 nmol/L) and in group paclitaxel (1 ng/mL) there were signifi-cant differences in growth inhibition, compared with control. While in group rapamycin(5 nmoL/L)+pacli-taxel(0.5ng/mL) there was significant difference in growth inhibition, compared with rapamycin (10 nmol/L) and paclitaxel (1 ng/mL) respectively. After cultured with rapamycin or paclitaxel alone, more tumor cells induced apoptosis than control. While after cultured with rapamycin and paclitaxel simultaneously, more tumor cells induced apoptosis than with rapamycin or paclitaxel alone. Conclusions Both rapamycin and paclitaxel had a good impressive effect on the three prostate cell lines (LNCaP-CA, LNCaP-C4-2, PC-3) with dose-dependent manner. After cultured with rapamycin and paclitaxel simultaneously, more tumor cells were induced apoptosis than with rapamycin or paclitaxel alone.

Objective To observe the mechanism of action of different extracts ofTangwang Mingmu Granules on high glucose induced VEGF and IL-1α gene and protein expressions in vascular endothelial cells.Methods Human umbilical vein endothelial cells EA.hy926 were divided into six groups: blank, high glucose,Tangwang Mingmu Granules, extract 1 (glycoside and flavonoid), extract 2 (organic acid and polysaccharides) and extract 3 (alkaloids) groups. High concentration glucose was used to establish the high glucose model in EA.hy926 cells. The expressions of VEGF and IL-1α mRNA were detected by semi-quantitative RT-PCR. The contents of VEGF and IL-1α protein were tested by ELISA.Results The gene expression and protein levels of VEGF and IL-1α were significantly up-regulated under the high glucose condition (P<0.05). However, the above indicators were significantly reduced after the treatment ofTangwang Mingmu Granules. The activity of different parts ofTangwang Mingmu Granules was as follows: extract 1> extract 3> extract 2.Conclusion The action intensity of glycosides and flavonoids, alkaloids, organic acids and polysaccharides on VEGF and IL-1α expression inTangwang Mingmu Granules weakens in sequence.

Aim The purpose of this study was to compare the differential expression of 15-lipoxygenase isoenzymes in the pulmonary arteries between normoxia and hypoxia and to explore their roles in the formation of hypoxic pulmomary vasoconstriction. Method Eighteen SD rats were randomly divided into two groups(n=9):the normoxic control group breathing fresh gas and the hypoxic group breeding in animal hypoxic incubator.Immunohistochemical method,in situ hybridization and Western blot were employed to determine certain 15-lipoxygenase isoenzymes which involved in the process of hypoxic pulmonary vasoconstriction.Results ①In normoxic control group,the expression of 15-LO-1 protein was detected in the pulmonary arteries;but the expression of 15-LO-2 protein wasn’t detected.②The expression of 15-LO-1 protein in hypoxic group was much stronger than that in normoxic group (P