Screening bioactive compounds from Danggui-shaoyao-san for treating sodium retention in nephrotic syndrome using bio-affinity ultrafiltration.

ETHNOPHARMACOLOGICAL RELEVANCE: Danggui-shaoyao-san (DSS), a representative formula of Traditional Chinese Medicine (TCM) for promoting blood circulation and diuresis (Huo-Xue-Li-Shui) therapy, has been used to clinically nephrotic syndrome (NS) and relieve nephrotic edema. AIM OF THE STUDY: To explore the effects and mechanisms of DSS in improving sodium retention and to identify the bioactive compounds from DSS. MATERIALS AND METHODS: DSS prescriptions were disassembled into Yangxue-Huoxue (YXHX) and Jianpi-Lishui (JPLS). A nephrotic rat model was induced with puromycin aminonucleoside (PAN), and the effects on urinary sodium excretion, urinary plasmin(gen) content, and plasmin activity of DSS, YXHX, and JPLS extracts were assessed. The inhibitory effects on urokinase-type plasminogen activator (uPA) and plasmin activity of extracts were evaluated in vitro. Bio-affinity ultrafiltration and high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (BAU-UPLC-Q/TOF-MS) were used to rapidly screen and qualitatively analyze the uPA/plasmin affinity compounds from DSS extract. Additionally, uPA/plasmin inhibition assays and molecular docking were used to verify the activity and affinity mechanisms of the potential bioactive compounds. RESULTS: In vivo, DSS, YXHX, and JPLS prevented sodium retention in nephrotic rats. DSS and YXHX treatment decreased urinary plasmin activity but did not alter urinary plasmin(ogen) concentration, and their extracts showed strong uPA and plasmin inhibitory activity in vitro. These results suggested that uPA and plasmin are direct targets of DSS and YXHX in intervening NS sodium retention. Using BAU-UPLC-Q/TOF-MS, gallic acids, methyl gallate, albiflorin, and 1,2,3,4,6-O-pentagalloylglucose (PGG) were screened as uPA or plasmin affinity compounds. Among them, PGG was found to be a uPA and plasmin dual inhibitor, with an IC50 of 6.861 µM against uPA and an IC50 of 149.0 µM against plasmin. The molecular docking results of PGG with uPA and plasmin were consistent with the verification results. CONCLUSION: Intervening in sodium retention by inhibiting uPA-mediated plasmin generation and plasmin activity in the kidneys could be possible mechanisms for DSS, as indicated by the results in PAN-induced nephrotic rats. We conclude that PGG is a potential bioactive compound responsible for the effect of DSS on natriuresis.

An effective treatment for cerebral hemorrhage: minimally invasive craniopuncture combined with urokinase infusion therapy.

OBJECTIVES: To evaluate and compare the curative effect between the minimally invasive craniopuncture combined with urokinase infusion therapy and the clearance of hematoma by craniotomy with small bone flap in treating patients with 30-80 ml hemorrhage in the basal ganglion part of the brain. METHODS: A multicenter, randomized control clinical trial was undertaken; it comprised of 22 hospitals in China. Three hundred and four patients with hemorrhage in the basal ganglion were randomly assigned to receive the craniopuncture combined with urokinase infusion therapy (n=159) or clearance of hematoma by craniotomy with small bone flap treatment (n=145). The main indexes of evaluation were the neurological impairment degree at day 14 after treatment, activities of daily living at day 90 and the case fatality by 90 days. RESULTS: The main results were as follows: (1) there was a significant difference in favorable outcomes (Barthel index >or=95) between the two groups (chi(2)=3.95, p<0.05), which showed a better prognosis in the craniopuncture group, although there was no significant difference in improving the neurological functions and activities of daily living at day 90; (2) there was a remarkable decrease in case fatality by 90 days in the cranipuncture group, with statistically significant difference between the two groups (chi(2)=5.35, p=0.02); (3) the re-bleeding rate in cranipuncture group was 8.8%, significantly (chi(2)=9.51, p=0.002) lower than 21.4% in the craniotomy group. CONCLUSION: The craniopuncture combined with urokinase infusion therapy could reduce the rate of re-bleeding after surgery and the case fatality by 90 days. It also could improve the activities of daily living (Barthel index >or=95) at day 90. Thus, this therapy was a safe and practical technique in treating cerebral hemorrhage (30-80 ml), especially suitable for hospitals in rural areas or developing countries.

Randomized adjuvant chemotherapy trial in high-risk, lymph node-negative breast cancer patients identified by urokinase-type plasminogen activator and plasminogen activator inhibitor type 1.

BACKGROUND: Most patients with lymph node-negative breast cancer are cured by locoregional treatment; however, about 30% relapse. Because traditional histomorphologic and clinical factors fail to identify the high-risk patients who may benefit from adjuvant chemotherapy, other prognostic factors are needed. In a unicenter study, we have found that levels of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor type 1 (PAI-1) in the primary tumor are predictive of disease recurrence. Thus, we designed the Chemo N(0) prospective randomized multicenter therapy trial to investigate further whether uPA and PAI-1 are such prognostic factors and whether high-risk patients identified by these factors benefit from adjuvant chemotherapy. After 4.5 years, we present results of the first interim analysis. METHODS: We studied 556 patients with lymph node-negative breast cancer. The median follow-up was 32 months. All patients with low tumor levels of uPA (< or = 3 ng/mg of protein) and of PAI-1 (< or = 14 ng/mg of protein) were observed. Patients with high tumor levels of uPA (> 3 ng/mg of protein) and/or of PAI-1 (> 14 ng/mg of protein) were randomly assigned to combination chemotherapy or subjected to observation only. All statistical tests were two-sided. RESULTS: A total of 241 patients had low levels of uPA and PAI-1, and 315 had elevated levels of uPA and/or PAI-1. The estimated 3-year recurrence rate for patients with low tumor levels of uPA and PAI-1 (low-risk group) was 6.7% (95% confidence interval [CI] = 2.5% to 10.8%). This rate for patients with high tumor levels of uPA and/or PAI-1 (high-risk group) was 14.7% (95% CI = 8.5% to 20.9%) (P = 0.006). First interim analysis suggests that high-risk patients in the chemotherapy group benefit, with a 43.8% lower estimated probability of disease recurrence at 3 years than high-risk patients in the observation group (intention-to-treat analysis: relative risk = 0.56; 95% CI = 0.25 to 1.28), but further follow-up is needed for confirmation. CONCLUSIONS: Using uPA and PAI-1, we have been able to classify about half of the patients with lymph node-negative breast cancer as low risk, for whom adjuvant chemotherapy may be avoided, and half as high risk, who appear to benefit from adjuvant chemotherapy.

[Clinical study on effect of sini decoction on ischemia/reperfusion injury by Holter monitoring in patients with acute myocardial infarction treated with thrombolytic therapy].

OBJECTIVE: To observe the clinical effect of Sini Decoction (SND) on ischemia/reperfusion injury in acute myocardial infarction (AMI). METHODS: Randomized case-control clinical trial was conducted to observe the change of Holter monitoring in 22 cases of AMI treated with thrombolytic therapy before and after treatment. RESULTS: The lasting time of acute ST segment, total burden of myocardial infarction, QRS score, QT dispersion and occurrence of reperfusion arrhythmia in patients received SND treatment were lower than those untreated with SND (P < 0.05). CONCLUSION: SND is helpful in improving reperfusion injury of thrombolytic therapy in AMI patients.

[Assessment of the anti-ischemic effect in patients with therapy refractory angina pectoris in end-stage coronary heart disease--results of chronic intermittent urokinase therapy]. / Antiischämischer Wirkungsnachweis bei Patienten mit therapierefraktärer Angina pectoris im Endstadium der koronaren Herzkrankheit--Ergebnisse zur chronisch-intermittierenden Urokinasetherapie.

Patients with refractory angina pectoris and end-stage coronary artery disease represent an increasing clinical problem. Numbers of these patients will increase in the future for improved survival due to effective secondary prevention of coronary artery disease. Next to the evaluation of clinical symptoms non-invasive objective parameters of myocardial ischemia are of major relevance before initiation of alternative treatment modalities and for verification of antiischemic effectiveness. Based on our own experience it can be shown that in these patients testing which is mainly based on the patients physical exercise capacity is only of limited value due to the early occurrence of clinical symptoms. Furthermore diffuse perfusion abnormalities reduce the sensitivity of electrocardiographic and scintigraphic detection of ischemic changes. In contrast indirect measures of ischemia relating to the systolic or diastolic function of the left ventricle like doppler-echocardiography and radionuclide ventriculography seem to be promising approaches. This is confirmed by the results from the application of long-term intermittent urokinase therapy. Long-term intermittent urokinase therapy leads to an absolute enhancement of myocardial perfusion, which makes this approach superior to other medical interventions which are mainly based on a reduction of cardiac work-load.

Adjuvant portal liver infusion in colorectal cancer with 5-fluorouracil/heparin versus urokinase versus control. Results of a prospective randomized clinical trial (colorectal adenocarcinoma trial I).

This prospectively randomized clinical trial was carried out in four Dutch hospitals to reduce the development of metachronous liver metastases and to get a better survival in patients with colorectal malignancies after surgically radical en bloc resection of the primary tumor and the regional lymph nodes. Three hundred seventeen patients were randomized to participate in three trial arms. One group of patients was treated by surgery alone (control group); in the other patients a catheter was placed in the dilated umbilical vein and advanced until the tip was lying in the left branch of the portal vein. Fifty percent of these patients got immediate postoperative portal infusion with 1 g 5-fluorouracil (5-FU) and 5000 U heparin daily for 7 days; the others received portal vein infusion with urokinase 10.000 U/hour for 24 hours only. Three hundred four patients were eligible. Overall hospital mortality was 3.6% (11 patients) and was not influenced by adjuvant treatment. After a median follow-up of 44 months 66 patients have died with relapse and 21 as a result of other causes. The chance of developing liver metastases and other distant metastases after portal infusion with 5-FU/heparin was one third of the chance in the control group (P less than 0.001). Only an insignificant reduction of the average death rate in the 5-FU/heparin group was found. In the urokinase group no significant effect in reducing metastases or in survival was noted. Before recommending cytotoxic portal infusion as an adjuvant treatment in patients with colorectal cancer, detailed analysis of other ongoing portal infusion studies has to be awaited and careful calculations have to be made regarding how many patients really can be saved by this treatment.