Expression of tissue factor in pancreatic adenocarcinoma is associated with activation of coagulation

AIM： To study expression of tissue factor （TF） in pancreatic cancer and its role in the development of thromboembolism.METHODS： TF expression was studied in eight human pancreatic carcinoma cell lines by Northern blot and indirect immunofluorescence. Expression of alternatively spliced TF （asTF） was assessed by RT-PCR. In addition, TF expression was determined by immunofluorescence in pancreatic tissues of 19 patients with pancreatic adenocarcinoma （PCa）, 9 patients with chronic pancreatitis （CP） and 20 normal controls. Plasma samples （30 PCa-patients, 13 CP-patients and 20 controls） were investigated for soluble TF levels and coagulation activation markers [thrombin-antithrombin Ⅲ complex （TAT）, prothrombin fragment 1 ＋ 2 （F1 ＋ 2）]. RESULTS： All pancreatic carcinoma cell lines expressed TF （8/8） and most of them expressed asTF （6/8）. TF expression at the protein level did not correlate with the differentiation of the carcinoma cell line. All but two pancreatic cancer tissue samples stained positive for TF （17/19）. In all samples of CP weak staining was restricted to pancreatic duct cells, whereas only a few subendothelial cells were positive in 9/20 of normal controls. TF and TAT levels in PCa patients were significantly elevated compared to controls whereas elevated F1 ＋ 2 levels did not reach statistical significance compared to controls. In CP patients TAT and F1 ＋ 2 levels proved to be significantly elevated compared to controls, although TAT elevation was less pronounced than in PCa patients. CONCLUSION： We conclude that in addition to the upregulated expression of TF on the cell membrane, soluble TF might contribute to activation of the coagulation system in pancreatic cancer.

COAGULATION ACTIVITY IN VARIOUS STRAINS OF YOSHIDA ASCITES HEPATOMA TISSUES AND CELLS OF RAT

To investigate the mechanism of chemotherapy related DIC, a quantitative and comparative study was carried out on the coagulation activities of tumor cells and tumor tissues in three strains of rat ascites hepatoma AH109A, AH272 and LY80. The content of thrombo-plastin was expressed as the relative concentration against that in the brain tissue. Ascites hepatoma cell and tissue of each strain obviously have different quantities of coagulants, such as tissue thromboplastin. Coagulation activities of tumor tissues are higher than that of tumor cells. The coagulation activities of tumor tissues and cells of AH272 were higher than that of AH109A and LY80. Our findings suggest that the initiation of DIC not only depend on the tissue thromboplastin, the other factors or the other mechanisms should be considered.

Chemical constituents and coagulation effects of the flowers of Rosa chinensis Jacq.

In this paper,the flowers of Rosa chinensis Jacq.were investigated and 14 compounds were isolated and identified,namely kaempferol(1),quercetin(2),isoquercitrin(3),afzelin(4),quercitrin(5),phenylethyl glucopyranoside(6),avicularin(7),juglanin(8),nicotiflorin(9),phenylethyl-6′-O-galloyl-β-D-glucopyranoside(10),tiliroside(11),methyl gallate(12),8-O-methylherbacetin-3-O-β-D-sophoroside(13),gallic acid(14).Among these compounds,compounds 7,9,12 and 13 were isolated from R.chinensis for the first time.These compounds and extracts of R.chinensis.were studied for coagulation activity in vitro.The results showed that tiliroside(11)had good effect on promoting blood coagulation,and the effect of tiliroside was better than that of Yunnanbaiyao.Juglanin(8)and nicotiflorin(9)could significantly shorten thrombin time(TT)and significantly elevated fibrinogen(FIB),which proved that juglanin and nicotiflorin had good procoagulant effect.

Chemical constituents from the Moutan Cortex charcoal and their potential coagulation activities

The chemical constituents of Cortex Moutan charcoal were studied in depth, and their coagulation activity was screened. The chemical constituents were isolated by polyamide, silica gel and Sephadex G-Sepharose chromatography purification, and their structures were identified by NMR spectroscopy and other analyses. A total of 10 compounds were obtained and identified as follows： 3-（2-furan）-l-（2-hydroxy-4-methoxy-phenyl）-2-propylene ketone （1）, 2-（2,5-hydroxy-4-methyl phenyl） propionate ethyl ester （2）, methyl tetradec-5-enoate （3）, 1,4-diethylcyclohexane （4）, catechol （5）, methyl-4-hydroxybenzoate （6）, 3-hydroxy- 2-methyl-4-pyrone （7）, 3,8-dihydroxy-2-metyl-chromone （8）, 3β-hydroxy-olean-12-en （9）, 1-monolinolein （10）. Among them, compounds 1 and 2 were new natural products, and 3-10 were isolated from the Cortex Moutan charcoal for the first time. The potential coagulation activities of compounds were evaluated, and compounds 2, 9 and 10 had strong hemostatic effects. While compounds 1 and 8 could significantly activate blood circulation.

Inter-chain disulfide bond improved protein trans-splicing increases plasma coagulation activity in C57BL/6 mice following portal vein FVIII gene delivery by dual vectors

Protein trans-splicing based dual-vector factor VIII （FVIII） gene delivery is adversely affected by less efficiency of protein splicing. We sought to increase the amount of spliced FVIII protein and plasma coagulation activity in dual-vector FVIII transgene in mice by means of strengthening the interaction of inteins, protein splicing elements, thereby facilitating protein trans-splicing. Dual-vector delivery of the FVIII gene in cultured cells showed that replacement of Met226 in the heavy chain and Asp1828 in the light chain with Cys residues could facilitate inter-chain disulfide linking and improve protein trans-splicing, increasing the levels of spliced FVIII protein. In this study, C57BL/6 mice were coadministered dual vectors of intein-fused human FVIII heavy chain and light chain with Cys mutations via portal vein injection into the liver. Forty-eight hours post-injection, plasma was collected and analyzed for FVIII antigen concentration and coagulation activity. These mice showed increased circulating FVIII heavy chain polypeptide （442± 151 ng mL i vs. 305±103 ng mL-1） and coagulation activi- ty （1.46±0.37 IU mL i vs. 0.85±0.23 IU mL-1） compared with control mice co-administered dual vectors expressing the heavy and light chains of wild-type FVIII. Moreover, coagulation activity was similar to that of mice receiving a single vector ex- pressing FVIII （1.79_＋0.59 IU mL-l）. These findings indicate that improving protein trans-splicing by inter-chain disulfide bonding is a promising approach for increasing the efficacy of dual-vector based FVIII gene transfer.

Coagulant compounds in Rubia cordifolia L.

Rubia cordifolia L.belongs to Rubiaceae family.Its medicinal parts are mainly root and rhizome,which is included in the Chinese Pharmacopoeia(2020),has the effect of cooling blood and stopping bleeding,removing stasis and channeling menstruation.However,studies on the hemostatic effect of R.cordifolia mainly focus on extracts,and the hemostatic components are less studied.Therefore,chemical constituents and coagulant activity of R.cordifolia were investigated in this paper.Eleven compounds were isolated and identified from R.cordifolia,including 1-hydroxy-2-methyl-9,10-anthraquinone(1),1,4-dihydroxy-2-methylanthraquinone(2),palmitic acid(3),tricosanoic acid(4),ω-hydroxypachybasin(5),ursolic acid(6),oleanolic acid(7),rubilactone(8),β-sitosterol(9),cordifoliol(10),1,3-dihydroxy-2-ethoxymethyl-9,10-anthraquinone(11).ω-Hydroxypachybasin was isolated from Rubia genus for the first time.1,4-Dihydroxy-2-methylanthraquinone,rubilactone and 1,3-dihydroxy-2-ethoxymethyl-9,10-anthraquinone could remarkably shorten the plasma calcium coating time in vitro,thus indicating that they had procoagulant activity.

Comparison of different combined treatment processes to address the source water with high concentration of natural organic matter during snowmelt period

The source water in one forest region of the Northeast China had very high natural organic matter（NOM） concentration and heavy color during snowmelt period. The efficiency of five combined treatment processes was compared to address the high concentration of NOM and the mechanisms were also analyzed. Conventional treatment can hardly remove dissolved organic carbon（DOC） in the source water. KMn O4pre-oxidization could improve the DOC removal to 22.0%. Post activated carbon adsorption improved the DOC removal of conventional treatment to 28.8%. The non-sufficient NOM removal could be attributed to the dominance of large molecular weight organic matters in raw water, which cannot be adsorbed by the micropore upon activated carbon. O3＋ activated carbon treatment are another available technology for eliminating the color and UV254 in water. However, its performance of DOC removal was only 36.4%, which could not satisfy the requirement for organic matter. The limited ozone dosage is not sufficient to mineralize the high concentration of NOM. Magnetic ion-exchange resin combined with conventional treatment could remove 96.2%of color, 96.0% of UV254 and 87.1% of DOC, enabling effluents to meet the drinking water quality standard. The high removal efficiency could be explained by the negative charge on the surface of NOM which benefits the static adsorption of NOM on the anion exchange resin. The results indicated that magnetic ion-exchange resin combined with conventional treatment is the best available technology to remove high concentration of NOM.