Hemodynamic characteristics of proper hepatic artery and portal vein in patients after splenectomy and devascularization / 临床肝胆病杂志

Objective To investigate the characteristics of hemodynamics of proper hepatic artery and portal vein after splenectomy and devascularization. Methods The clinical data of 103 patients with portal hypertension who underwent splenectomy and devascularization in the Capital Medical University-Affiliated You'an Hospital from April 2014 to February 2019 were retrospectively analyzed. Their hemodynamics of the proper hepatic artery and portal vein were recorded before and 1 week-, and 1-, 3-, 6-, 12-, and 24-months after surgery and then statistically analyzed. Continuous data with normal distribution were compared using paired-samples t test. Results Compared with the before surgery data, the portal vein diameter, portal vein flow, maximum velocity, and average velocity of the portal vein were all significantly decreased 1-week-, 1-, 3-, 6-, 12-, and 24-months after splenectomy and devascularization (all P < 0.05). The blood flow and velocity of the proper hepatic artery was significantly increased 1 week and 1 month after surgery (all P < 0.05); however, there was no statistically significant difference at 3-, 6-, 12-, and 24-months after surgery. Conclusion The diameter, flow, and flow velocity of the portal vein after splenectomy and devascularization were significantly lower than those before surgery, whereas the proper hepatic artery flow and flow velocity were increased within 1 month after surgery and then returned back to the pre-surgery levels 3 months after surgery.

In vitro and in vivo evaluation of folate-mediated triptolide exosome targeted delivery system against malignant melanoma / 药学学报

In this study, exosomes were extracted from human malignant melanoma cell A375. Folic acid (FA) receptor was used as target and triptolide (TPL) was used as model drug to prepare exosome targeted drug delivery system, FA-Exo/TPL. The physicochemical properties and antitumor effect were evaluated in vivo and in vitro. Gradient centrifugation method was applied to collect exosomes. Then, exosome was modified with FA for loading TPL. The particle sizes of the FA-Exo/TPL were about 100 nm with a double-layer membrane structure like a tray. It is characteristic of high encapsulation efficiency and drug loading. In vitro experiments showed that FA-Exo/TPL could be effectively uptaken by A375 cells, thus significantly inhibiting proliferation and promoting apoptosis the cells. In vivo experiment results showed that FA-Exo/TPL could effectively inhibit the growth of tumor tissue, prolong the model mice life cycle, and significantly reduce the systemic toxicity of the free drug, playing a synergistic and toxic role. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of Fudan University Shanghai Cancer Center. This study provides a new strategies and methods for the preparation of TPL against malignant melanoma.

Research advances in exosomes as biomarkers for tumor diagnosis / 医学研究生学报

Malignancy is one of the leading causes of death worldwide. Effective screening and early diagnosis of tumors are clinically difficult. The existing tumor markers have low sensitivity and poor specificity. Exosomes, the nanoscale vesicles with a phospholipid bilayer structure, are widely distributed in various body fluids. Studies have shown that tumor-derived exosomes (TEXs) are closely related to the occurrence and progression of cancers. The contents of TEXs including proteins, RNA and DNA, glycoproteins, glycolipids and lipids can serve as highly sensitive tumor-specific markers, playing a crucial role in the basic research and clinical examination of cancers. Therefore, TEXs are expected to become new non-invasive tumor diagnostic biomarkers. This review describes the biological characteristics of exosomes, their advantages as diagnostic biomarkers and their applications in diagnosis of tumors, treatment monitoring and prognosis evaluation.

Predicting value of PDGF-BB combined with CLIF-COFs for the occurrence of ARDS in post-liver transplantation patients / 国际外科学杂志

Objective To investigate the role of cytokines combined with CLIF consortium organ failure score (CLIF-COFs) for predicting the occurrence of acute respiratory distress syndrome (ARDS) in for post-liver transplant for hepatitis B-related acute-on-chronic liver failure (HB-ACLF) patients.Methods From Jul.2014 to Oct.2017,there were 37 cases of HB-ACLF undergoing liver transplantation in Beijing YouAn Hospital,Capital Medical University.According to whether the patients happened ARDS or not,37 cases were divided into ARDS group (n =9) and non-ARDS group (n =28).All patients' plasma was prospectively collected immediately before liver transplantation and on the I st,3rd,5th,7th day post-liver transplantation.The serum levels of twenty-seven cytokines were determined by 200 LUMINEX liquid chip technology.Cytokines,CLIF-COFs,clinical and biochemical indexes were analyzed with logistic regression and the receiver operating characteristic (ROC) to confirm the correlation with ARDS post liver transplantation.Results There were significant differences between HB-ACLF patients between ARDS group and non-ARDS group in age and pre-transplant infection (P ＜ 0.05).The CLIF-COFs was higher in ARDS non-without than that in non-ARDS group (P =0.019).The serum levels of vascular endothelial growth factor and platelet-derived growth factor bb were lower but IL-6 was higher post transplantation in ARDS group.The COX analysis indicated that CLIF-COFs and post liver transplantation PDGF-BB were predictors of post-LT ARDS.The area under the receiver operating characteristic (AUROC) curves was 0.728 and 0.175,respectively.The area under the curve of the discriminatory power of CLIF-COFs combined with PDGF-BB was 0.913,and the maximum Youden index is 0.786.Conclusion CLIF-COFs combines with PDGF-BB can predict the occurrence of ARDS post-liver transplantation in HB-ACLF patients.

The impact of splenectomy and esophagogastric devascularization on the nutrition status of patients with cirrhosis and portal hypertension / 中华肝胆外科杂志

Objective To study the impact of splenectomy and esophagogastric devascularization on the nutritional status of patients with cirrhosis and portal hypertension.Methods Sixty consecutive patients with cirrhosis and portal hypertension who underwent splenectomy and esophagogastric devascularization at the Beijing YouAn Hospital from April 5,2015 to January 23,2017 were included in this study.The body mass index (BMI),albumin (Alb),prealbumin (PA) and lymphocyte counts were prospectively collected at the end of 1-week,1-month,3-month,6-month and 1-year after surgery.The postoperative results were compared with the preoperative results in these patients.Results The BMI results obtained at 1-week and 1-month after surgery were significantly lower than the preoperative level [(22.14 ± 3.08)kg/m2 vs.(22.85 ± 3.14) kg/m2,(21.72 ± 3.05) kg/m2 vs.(22.86 ± 3.16) kg/m2,P ＜ 0.05].The BMI result at the end of 1-year after surgery was significantly elevated when compared with the preoperative level [(23.24 ± 3.64) kg/m2 vs.(22.68 ± 3.47) kg/m2,P ＜ 0.05].The ALB levels at 1-month and 3-month after surgery were significantly higher than the preoperative level [(39.87 ± 4.22)g/L vs.(35.35 ±5.15) g/L,(39.35 ± 4.75) g/L vs.(34.82 ± 5.50) g/L,P ＜ 0.05].The PA obtained at 1-week after surgery was significantly lower than the preoperative levels [(79.59 26.52)mg/L vs.(121.77 ±39.96)mg/L,P ＜ 0.05].The lymphocyte counts at all the points after surgery were significantly higher than the preoperative level (P ＜ 0.05).Conclusion Short term and long term nutritional status improved in patients with cirrhosis and portal hypertension after splenectomy and esophagogastric devascularization.

Effect of atorvastatin on advanced glycation end products induced monocyte chemoattractant protein-1 expression in cultured human endothelial cells / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of atorvastatin on advanced glycation end products (AGE) induced monocyte chemoattractant protein-1 (MCP-1) expression in human umbilical vein endothelial cells (HUVECs) and whether this effect could be linked to peroxisome proliferator-activated receptor-γ (PPAR-γ) and nuclear factor-κB (NF-κB).</p><p><b>METHODS</b>Grouping: (1) Blank control group; (2) BSA group; (3) AGE group: cells were incubated with different concentrations of AGE (10(-4), 10(-3), 10(-2) and 10(-1) g/L) for 24 hours; (4) AGE + Atorvastatin group: cells were incubated with different concentrations of atorvastatin (0.1, 1, 10 µmol/L) for 1 hour, then incubated with AGE (10(-1) g/L) for 24 hours; (5) PPAR-γ agonist (15 d-PGJ2) group: cells were incubated with 15 d-PGJ2 (10 µmol/L) for 1 hour, then incubated with AGE (10(-1) g/L) for 24 hours; (6) PPAR-γ inhibitor (GW9662) group: cells were incubated with GW9662 (5000 nmol/L) for 1 hour, then incubated with atorvastatin (1 µmol/L) and AGE (10(-1) g/L) for 24 hours. Collagenase was used to isolate the endothelial cell from human umbilical vein; RT-PCR was performed to examine the mRNA expression of MCP-1 and PPAR-γ; Western blot was performed to detect NF-κB p65 protein.</p><p><b>RESULTS</b>(1) The expression of MCP-1 mRNA was increased in proportion with increasing concentrations of AGEs which could be blocked by atorvastatin in a dose-dependent manner. (2) AGE (10(-1) g/L) significantly downregulated the expression of PPAR-γ mRNA (0.22 ± 0.08 vs. 0.69 ± 0.09, P < 0.01) while upregulated the expression of phospho-NF-κB p65 protein (0.78 ± 0.06 vs. 0.31 ± 0.01, P < 0.01) and nonphospho-NF-κB p65 protein (1.61 ± 0.16 vs. 0.59 ± 0.14, P < 0.01) compared with the control group which could be significantly attenuated by atorvastatin. (3) PPAR-γ agonist decreased the expression of phospho-NF-κB p65 protein (0.21 ± 0.01 vs. 0.78 ± 0.06, P < 0.01), nonphospho-NF-κB p65 protein (0.67 ± 0.14 vs. 1.61 ± 0.16, P < 0.01) and MCP-1 mRNA (0.17 ± 0.02 vs. 0.93 ± 0.12, P < 0.01) compared with AGE (10(-1) g/L) group. (4) PPAR-γ inhibitor antagonized the effect of atorvastatin on the expression of phospho-NF-κB p65 protein, nonphospho-NF-κB p65 protein and MCP-1 mRNA stimulated by AGE in HUVECs (P < 0.01).</p><p><b>CONCLUSION</b>The anti-inflammatory properties of atorvastatin in AGE stimulated HUVECs may partly be attributed to the effect on upregulation of PPAR-γ and downregulation of NF-κB signaling pathway.</p>

Effects of human tissue kallikrein gene transfer on the migration of vascular smooth muscule cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of adenovirus-mediated human tissue kallikrein (Ad-hKLK1) gene transfer on platelet-derived growth factor-BB (PDGF-BB)-induced migration of vascular smooth muscle cells from spontaneously hypertensive rats (VSMC(SHR)).</p><p><b>METHODS</b>A bicistronic recombinant adenovirus vector (Ad-hKLK1) carrying the target hKLK1 gene and the reporter gene EGFP was constructed. VSMCs isolated from the thoracic aorta of male SHR were passaged, and the quiescent VSMC(SHR) in passages 3-6 seeded in 6-well plates were treated with Ad-hKLK1 and control virus. Human PDGF-BB or icatibant Hoe140, a BK B2 antagonistat, was used as the chemoattractant and placed in the bottom chamber of the Boyden chamber. The mRNA expressions of bradykinin B1 receptor and B2 receptor were detected by RT-PCR in VSMC(SHR).</p><p><b>RESULTS</b>hKLK1 gene transfer significantly inhibited PDGF-BB-induced migration of VSMC(SHR), with the peak inhibition rate of 34.6% (P<0.001). PDGF-BB significantly increased the mRNA expression of B2 receptor but not B1 receptor in VSMC(SHR).</p><p><b>CONCLUSIONS</b>hKLK1 gene transfer can inhibit the migration of VSMC(SHR) induced by PDGF-BB, and the inhibitory effects may be not mediated by bradykinin B2 receptor.</p>

Effects of human tissue kallikrein 1 gene delivery on carotid artery neointima formation after balloon angioplasty in spontaneously hypertensive rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of human tissue kallikrein 1(Ad-hKLK1) gene delivery on the neointima formation in carotid arteries of spontaneously hypertensive rats (SHRs).</p><p><b>METHODS</b>Carotid artery restenosis was induced in male SHR rats by balloon-injury. Rats were randomly assigned into 4 groups: Sham-operated (n = 6); Angioplasty (phosphate buffered solution 50 microl, n = 8); Vector virus (control virus, 1 x 10(9) IU in 50 microl, n = 8) and Ad-hKLK1(Ad-hKLK1, 1 x 10(9) IU in 50 microl, n = 8). Rats were sacrificed 4 weeks later. The wall-to-lumen area ratio and intima/media ratio in carotid artery were assessed by image analysis in HE stained sections. The mRNA bradykinin receptor (B1R and B2R) expressions were detected by RT-PCR. The protein expression of the cycle-independent kinase inhibitors p27Kip1 and p2lCip1 were determined by Western blot analysis.</p><p><b>RESULTS</b>Wall-to-lumen area ratio reduced 35.6% and intima/media ratio reduced 38.8%in Ad-hKLK1 treated SHRs compared to angioplasty group (all P < 0.001). The expression of p27Kip1 and p2lCip1 increased significantly in Ad-hKLK1 treated SHRs compared with angioplasty rats (all P < 0.001). The mRNA expression of B2R was significantly upregulated in angioplasty rats compared with sham-operated rats (P < 0.05) while mRNA expression of B1R was similar between the 2 groups.</p><p><b>CONCLUSION</b>hKLK1 gene delivery may effectively reduce neointimal formation via downregulating bradykinin B2R and up-regulating the expressions of p27Kip1, p2lCip1 signaling pathways in carotid arteries of SHRs after balloon injury.</p>

Effects of human tissue kallikrein gene delivery on the proliferation of vascular smooth muscle cells / 中华心血管病杂志

<p><b>OBJECTIVE</b>Tissue kallikrein cleaves kininogen substrate to produce vasoactive kinin peptides that have been implicated in the proliferation of vascular smooth muscle cells. We investigated the effects of adenovirus-mediated human tissue kallikrein (Ad-hKLK1) gene delivery on the proliferation of vascular smooth muscle cells of SHR (VSMCs(SHR)) induced by platelet derived growth factor-BB (PDGF-BB).</p><p><b>METHODS</b>Primary VSMCs(SHR) were isolated and cultured from thoracic aorta of male SHR. The VSMCs(SHR) proliferation induced by PDGF-BB was accessed by cell counting and methyl thiazolyl tetrazolium (MTT). Western blot was used to determine the protein expression of hKLK1, the cycle-independent kinase inhibitors p27(Kip1) and p21(Cip1). The mRNA expressions of bradykinin B1 receptor and B2 receptor were detected by RT-PCR in VSMCs(SHR).</p><p><b>RESULTS</b>Proliferation of VSMCs(SHR) induced by PDGF-BB was significantly inhibited post transfection of Ad-hKLK1 (20-100 MOI) in a MOI-dependent manner. The peak inhibition titer of Ad-hKLK1 was 100 MOI with peak inhibition rate of 39.3% (cell counting, n = 3, P < 0.01), 30.2% (MTT, n = 3, P < 0.01) and 36.4% (peak stunning rate of cell-cycle in phase G(0)/G(1)). The inhibitory effects of proliferation and cell-cycle caused by hKLK1 gene delivery could be abolished by Hoe140, a bradykinin B2 receptor antagonist. The protein expression of p27(Kip1) and p21(Cip1) increased significantly after the hKLK1 gene delivery, whereas Hoe140 nearly completely blocked these effects (n = 3, P < 0.001, respectively). PDGF-BB also significantly upregulated the mRNA expression of B2 receptor but not B1 receptor in VSMCs(SHR).</p><p><b>CONCLUSION</b>The hKLK1 gene delivery could inhibit PDGF-BB induced proliferation in VSMCs(SHR) through Bradykinin B2 receptor and up-regulate expression of p27(Kip1) and p2l(Cip1).</p>

Purification and Characterization of Laccase from Monodictys asperospera (Cooke & Massee) Ellis / 微生物学通报

A new wood-degrading fungus Monodictys asperospera(Cooke & Massee) Ellis with a high level of laccase production was chosen to study.This laccase was purified by ammonium sulfate precipitation,DEAE-cellulose and sephacryl S-300.Purification of about 8.1 fold was achieved with an overall yield of 5.7%.Its molecular weight was estimated to be about 77 kD.The optimum temperature and pH of the lac-case activity were 55?C and 6.0,respectively.Kinetic studies of the laccase showed that the Km and the Vmax for using syringaldazine as substrate was 0.163 mmol/L and 0.194 mmol/(L.min),respectively.The carbo-hydrate content was 18.14%.In addition,it was found that laccase activity was significantly inhibited by Cu2+.