In the progression process of liver diseases, pathological changes occur in the vascular endothelium, platelets, coagulation, anticoagulation, fibrinolysis systems of patients, and "rebalancd haemostasis" is formed. The coexistence of hypocoagulation and hypercoagulation tendency in the peripheral blood of patients made the laboratory testing data complex and difficult to interpret, especially prothrombin time and activated partial thromboplastin time could not truly reflect the net effect of haemostasis disorder in patients with liver diseases, which often led to misjudgment and overtreatment by doctors and led to adverse clinical consequences. In recent years, thromboelastography, rotational thromboelastometry and thrombin generation assay had been used to assess bleeding risk in patients with liver diseases and had made progress in transfusion management. On the other hand, von Willebrand factor and thrombospondin type 1 motif member 13 has been shown to be closely related to the progression of liver diseases and could warn the adverse clinical outcomes effectively.

In the progression process of liver diseases, pathological changes occur in the vascular endothelium, platelets, coagulation, anticoagulation, fibrinolysis systems of patients, and "rebalancd haemostasis" is formed. The coexistence of hypocoagulation and hypercoagulation tendency in the peripheral blood of patients made the laboratory testing data complex and difficult to interpret, especially prothrombin time and activated partial thromboplastin time could not truly reflect the net effect of haemostasis disorder in patients with liver diseases, which often led to misjudgment and overtreatment by doctors and led to adverse clinical consequences. In recent years, thromboelastography, rotational thromboelastometry and thrombin generation assay had been used to assess bleeding risk in patients with liver diseases and had made progress in transfusion management. On the other hand, von Willebrand factor and thrombospondin type 1 motif member 13 has been shown to be closely related to the progression of liver diseases and could warn the adverse clinical outcomes effectively.

Background: Heterologous synthesis presents a promising new approach for accessing the active ingredients of traditional Chinese medicine (TCM), contributing to the conservation of natural medicinal resources. Compound Danshen preparation, a widely used TCM formulation, is designed to treat coronary heart disease and angina pectoris. It is primarily composed of Salviae Miltiorrhizae Radix et Rhizoma (Danshen), Notoginseng Radix et Rhizoma (Sanqi), and borneol (Bingpian). Danshen primarily yields tanshinones and phenolic acids, while Sanqi produces notoginsenosides. Borneol serves as an auxiliary agent to promote mental clarity, dissipate heat, and relieve pain. Objectives: The objective is to employ heterologous synthesis in a single yeast strain to produce the active ingredients of Compound Danshen preparation, and these include notoginsenosides, tanshinones, and borneol. Methods: Firstly, the "plug-and-play" terpene synthase screening framework in Saccharomyces cerevisiae was used for the identification of a novel, highly efficient bornyl diphosphate synthase. Furthermore, by leveraging the Compound Danshen preparation as a basis to concurrently reconstruct the biosynthetic pathways for the notoginsenoside precursor protopanaxadiol (a triterpene), the tanshinone precursor miltiradiene (a diterpene), and borneol (a monoterpene) within a single yeast strain. Results: This engineered strain, termed Compound Danshen Yeast 1.0, successfully produced protopanaxadiol at 62.34 mg/L, miltiradiene at 15.38 mg/L, and borneol at 2.54 mg/L in shake-flask fermentation. Conclusions: This research signifies the inaugural cross-species and multicomponent synthetic biology endeavor that enables the synthesis of active ingredients in engineered cells, setting the stage for the industrial manufacture of TCM compounds.

Objective:To investigate the performance of von willebrand factor antigen (vWF:Ag) and D-dimer in predicting thrombotic risk in nonvalvular atrial fibrillation (NVAF) patients with anticoagulant therapy.Methods:From March 2017 to March 2019, 256 patients were enrolled, including 152 males and 104 females, aged (57.9±20.4) years old; according to the end-point events during the follow-up period, the patient group was further divided into 227 cases in the no-event group and 29 cases in the thrombotic event group;50 cases in the control group, including 30 males and 20 females, aged (45.0±5.3) years old. vWF:Ag was detected by blood coagulation instrument and determination of D-dimer was done by fluor-euzyme linked immunoassay Analyzer. Mann-Whitney U test was used for data comparison between any two groups, Kruskal-Wallis H test was used for comparison among multiple groups and multivariate correlation analysis was done by Logistic regression to obtain odds ratio ( OR). The prediction performance with thrombotic events of vWF:Ag and D-dimer was evaluated by ROC curve, Kaplan-Meier curve was used to analyze the survival curve and the hazard ratio ( HR) was obtained by Cox proportional hazard regression model. Results:The levels of vWF:Ag and D-dimer in the control group were 103% (86%-131%) and 249 (90-522) μg/L, 234% (102%-623%) and 744 (100-3 352) μg/L in the patient group; in the patient group, of which 225% (102%-623%) and 650 (100-3 281) μg/L in non-event group, 333% (210%-494%) and 1 325 (487-3 352) μg/L in thrombus event group; compared the healthy control, the levels of vWF:Ag and D-dimer were increased in patients group ( P<0.001), of which non-event groups were higher than healthy controls ( P<0.001), and the thrombotic event group was higher than that of the non-event group ( P<0.001). Plasma vWF:Ag level and D-dimer level in NVAF patients were higher than those in the control group ( P<0.001). Plasma vWF:Ag level and D-dimer level in the non-event group were significantly higher than those in the healthy control group ( P<0.001). The plasma vWF:Ag and D-dimer levels of patients in the thrombotic event group were significantly higher than those in the non-event group patients ( P<0.001). The result of ROC showed that the critical value of vWF: Ag for predicting thrombosis within 3 months of NVAF patients was 229% and area under the curve (AUC) was 0.839 (95% CI:0.784-0.894); When the critical value of D-dimer was 588 ng/ml, AUC was 0.803 (95% CI:0.745-0.861).While vWF:Ag combined with D-dimer, AUC was 0.868 (95% CI:0.826-0.909). Logistic regression analysis showed that plasma vWF:Ag level in NVAF patients was significantly correlated with age ( OR=10.240, 95%CI 2.773-37.820), congestive heart failure ( OR=34.779, 95%CI 8.010-151.019), hypertension ( OR=0.068, 95%CI 0.023-0.198) and type 2 diabetes ( OR=6.618, 95%CI 2.469-17.734) ( P<0.001), as well as was significantly correlated with vascular disease ( OR=4.801, 95%CI 1.204-19.145) ( P=0.026). Plasma D-dimer level was significantly correlated with congestive heart failure ( OR=0.146, 95%CI 0.036-0.588) and medication compliance ( OR=0.114, 95%CI 0.016-0.832) ( P value was 0.007 and 0.032). Survival analysis showed that the cumulative probability of thrombosis within 3 months was significantly increased (Log-rank χ2 was 11.394, 17.895 and 32.825 respectively, P value<0.001) in the patients with plasma levels above the critical value of vWF:Ag, D-dimer or vWF:Ag combined with D-dimer. Cox proportional regression model showed that neither vWF:Ag nor D-dimer could independently predict thrombotic events during anticoagulant therapy( HR was 0.866 and 0.834, P-value was 0.253 and 0.152, respectively), but it could improve the prediction performance significantly( HR=0.780, P=0.048) for combined application of both vWF:Ag and D-dimer. Conclusion:The changes with plasma vWF:Ag and D-dimer levels in NVAF patients were associated with a variety of clinicopathological factors and closely related to the risk of thrombosis within 3 months. Combined application could provide the effective basis for clinical prediction of the condition.

Objective To investigate the effect of SKP2 expression on radiation induced bystander effect (RIBE) of esophageal cancer cells.Methods The esophageal cancer cell lines with different SKP2 levels were applied for the study and the SKP2 expression was identified by Western blot.Micronuclei (MN) assay and DNA foci assay were used to evaluate the effect of SKP2 on RIBE.The cells were transfected with SKP2 gene or SKP2 siRNA to further verify the effect of SKP2 on RIBE.Results MN assay showed that the bystander effect induced by the cells with a high level of SKP2 was lower than that induced by the cells with a lower level of SKP2 (t =8.06,P ＜ 0.01).These results were further confirmed by the gene transfection experiments.When the expression of SKP2 was increased,RIBE was decreased (t=11.12,10.16,P ＜ 0.01).Contrarily,when the expression of SKP2 was reduced,RIBE was increased (t =8.39,8.83,P ＜ 0.01).γ-H2AX foci formation assay disclosed that when SKP2 expression in the irradiated cells increased,the repair ability of DNA damage in the bystander cells was higher than the control (t =6.85,7.10,P ＜ 0.01).With the expression of SKP2 decreased,the repair ability of DNA damage was lower than the control (t =7.66,8.47,P ＜ 0.01).Conclusions Over-expression of SKP2 inhibits RIBE of esophageal cancer cells,at least partly through regulating DNA damage repair ability.

OBJECTIVETo study synthesis of baicalin-copper and baicalin-aluminium complex and its antimicrobial, anti-tumor activity and anti-tumor effect against macrophages.

METHODBaicalin was reacted with metallic salt under a weak base condition to produce baicalin-copper and baicalin-aluminium complex. Baicalin and its synthesized complex were detected for antimicrobial activity against Staphylococcus aureus, Hay bacillus, Escherichia coli, Salmonella and Candida albicans by twofold broth dilution technique. Their anti-tumor activity against A549 and IC50 of HepG2 cells and anti-tumor effect against macrophages were detected by the MTT. And their phagocytic effect on macrophages was determined by the neutral red assay.

RESULTThe yields of baicalin-copper and baicalin-aluminium complex were 73.93% and 91.08%, respectively. The minimum inhibitory concentration (MIC) value against Staphylococcus aureus, Hay bacillus, Escherichia coli, Salmonella and Candida albicans was 0.0004, 0.0009, 0.0004, 0.0009, 0.000 4 mol x L(-1) for baicalin-copper complex and 0.0011, 0.0011, 0.0011, 0.0011, 0.0005 mol x L(-1) for baicalin-aluminium complex. The IC50 values against A549 and HepG2 cells were 89.6, 22.6 micromol x L(-1) for baicalin-copper complex, and 138.8, 97.2 micromol x L(-1) for baicalin-aluminium complex. The inhibitory ratio of macrophage on A549 cell was 43.52%, 80.89%, 52.66%, respectively, after the macrophages were stimulated by baicalin, baicalin-copper and baicalin-aluminium complex at a concentration of 160 micromol x L(-1).

CONCLUSIONThe acute toxicity test in mice showed that the complex was nontoxic to mice. Baicalin-copper complex showed the highest antimicrobial, anti-tumor activity, and the strongest effect on the anti-tumor activity of macrophage, while baicalin showed the lowest activities compared with baicalin-copper and baicalin-aluminium complex.

Aluminum ; Animals ; Anti-Bacterial Agents ; chemical synthesis ; chemistry ; pharmacology ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Copper ; Drugs, Chinese Herbal ; chemical synthesis ; chemistry ; pharmacology ; Flavonoids ; Humans ; Mice ; Microbial Sensitivity Tests

The flavonoid-metal complexes showed obviously stronger bioactivities such as antibiosis, antivirus, anti-inflammatory, anti-tumor and anti-free-radical, possibly because of the stronger binding force caused by the change in complex structure and accessibility to target spots, or the synergy effect between flavonoids and metallic ions. This essay summarizes studies on bioactivity and mechanism of flavonoid-metal complexes, in order to provide reference for in-depth study and development on effective constituents contained in flavonoid traditional Chinese medicines.
Animals ; Coordination Complexes ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Flavonoids ; pharmacology ; Humans ; Medicine, Chinese Traditional

OBJECTIVETo investigate the effect of ceftiofur hydrochloride on the pharmacokinetics of matrine in rats.

METHODThe rats were divided into two groups: one group was administrated with matrine only (control group) and the other was administrated with matrine in combination with ceftiofur hydrochloride. HPLC-UV method was used for determining the plasma concentration of matrine in both groups. The pharmacokinetic parameters were calculated from the plasma concentration-time data using the DAS 2. 1. 1 software program.

RESULTThe main pharmacokinetic parameters for the control group were C(max) = 21.113 9 mg x L(-1), T(max) = 0.75 h, t1/2alpha = 1.34 h, t1/2beta = 3.509 h, AUC(0-t) = 90.984 mg x h(-1) x L(-1) and AUC(0-inifinity) = 100.346 mg x h(-1) x L(-1), and the data for the combination group were C(max) = 11.707 mg x L(-1), T(max) = 0.917 h, t1/2alpha = 1.598 h, t1/2beta = 3.247 h, AUC(0-t) = 53.28 mg x h(-1) x L(-1) and AUC(0-inifinity) = 60.035 mg x h(-1) x L(-1).

CONCLUSIONThe plasma concentration of matrine and bioavailability in combination group were significantly lower than those of the control group. In combination group, matrine had a higher clearance and volume of distribution in the central compartments, as well as a lower volume of distribution in the peripheral compartments.

Alkaloids ; administration & dosage ; blood ; pharmacokinetics ; Animals ; Cephalosporins ; administration & dosage ; blood ; Drug Interactions ; Male ; Quinolizines ; administration & dosage ; blood ; pharmacokinetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the pharmacokinetics of matrine (MT) intramuscular administration in rat.

METHODPlasma concentration of matrine was determined by HPLC under the following conditions: column (Shim-pack VP-ODS, 4. 6 mm x 150 mm, 5 m); eluent (acetonitrile-0.02 mol ammonium acetate buffer-triethylamine 30: 70: 0.04); flow rate was 1 mL x min(-1) and ultraviolet detection wavelength was set at 220 nm; column temperature 40 degrees C; aliquot injected 20 microL. All data of concentration-time of matrine were treated with pharmacokinetics program DAS 2. 1. 1.

RESULTA simple, sensitive and reliable method for determining matrine in rat plasma by HPLC was established. The plasma concentration time profiles of MT fitted in with two-compartment models well, and the main pharmacokinetic parameters found for MT after i. m. infusion were as follows: C(max) = 21.113 9 mg x L(-1), t(max) = 0.75 h, t1/2alpha 1.34 h, t1/2beta = 3.509 h, AUC(0-t) = 90.984 mg x h(-1) x L(-1), AUC(0-infinity) = 100.346 mg x h(-1) x L(-1).

CONCLUSIONCompare with oral administration, the matrine is absorbed well and distributes fast with intramuscular administration; the absolute bioavailability of matrine is higher. According to this, the pharmacological action is also stronger and duration is longer.

Alkaloids ; administration & dosage ; pharmacokinetics ; Animals ; Chromatography, High Pressure Liquid ; methods ; Female ; Injections, Intramuscular ; Male ; Quinolizines ; administration & dosage ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley