A double-blind, placebo-controlled, randomized, multicenter study to investigate CHInese Medicine Neuroaid Efficacy on Stroke recovery (CHIMES Study).

UNLABELLED: Rationale Traditional Chinese Medications(TCM) have been reported to have beneficial effects in stroke patients, but were not rigorously evaluated by GCP standards. Aim This study tests the hypothesis that Neuroaid, a TCM widely used in China post-stroke, is superior to placebo in reducing neurological deficit and improving functional outcome in patients with acute cerebral infarction of an intermediate severity. Design This is a multicenter, randomised, double-blind, placebo-controlled study of Neuroaid in ischemic stroke patients with National Institute of Health Stroke Scale(NIHSS) 6-14 treated within 48 h of stroke onset. Neuroaid or placebo is taken (4 capsules) 3 times daily for 3 months. Treatments are assigned using block randomization, stratified for centers, via a central web-randomization system. With a power of 90% and two-sided test of 5% type I error, a sample size is 874. Allowing for a drop-out rate of up to 20%, 1100 individuals should be enrolled in this study. Study Outcomes The primary efficacy endpoint is the modified Rankin Scale(mRS) grades at 3 months. Secondary efficacy endpoints are the NIHSS score at 3 months; difference of NIHSS scores between baseline and 10 days, and between baseline and 3 months; difference of NIHSS sub-scores between baseline and 10 days, and between baseline and 3 months; mRS at 10 days, 1 month, and 3 months; Barthel index at 3 months; Mini Mental State Examination at 10 days and 3 months. Safety outcomes include complete blood count, renal and liver panels, and electrocardiogram. STUDY REGISTRATION: ClinicalTrials.gov identifier: NCT00554723.

Polymorphisms in VEGF and IL-8 predict tumor recurrence in stage III colon cancer.

BACKGROUND: Identifying molecular markers for tumor recurrence is critical in successfully selecting patients with stage III colon cancer who are more likely to benefit from adjuvant chemotherapy. The present study analyzed a subset of 10 polymorphisms within eight genes involved in the tumor angiogenesis pathway and their impact on prognosis in stage III colon cancer patients treated with adjuvant chemotherapy. PATIENTS AND METHODS: Blood samples were obtained from 125 patients with locally advanced colon cancer at University of Southern California medical facilities. DNA was extracted from peripheral blood and the genotypes were analyzed using PCR-restriction fragment length polymorphism and 5'-end [gamma-(33)P] ATP-labeled PCR protocols. RESULTS: Polymorphisms in vascular endothelial growth factor (VEGF) (C+936T; P = 0.003, log-rank test) and interleukin-8 (IL-8) (T-251A; P = 0.04, log-rank test) were independently associated with risk of recurrence in stage III colon cancer patients. In combined analysis, grouping alleles into favorable versus nonfavorable alleles, high expression variants of VEGF C+936T and IL-8 T-251A were associated with a higher likelihood of developing tumor recurrence (P < 0.001). CONCLUSION: High expression variants of VEGF C+936T and IL-8 T-251A were associated with shorter time to tumor recurrence, indicating that the analysis of angiogenesis-related gene polymorphisms may help to identify patient subgroups at high risk for tumor recurrence.

Growth inhibition and induction of apoptosis in U937 cells by Coptis chinensis extract.

Dried Coptidis Rizoma was extracted with boiling water. Conditioned medium was prepared by stimulating human peripheral blood mononuclear cells with Coptidis Rizoma extract (CR). The conditioned medium was then added to human leukemic U937 cells suspension for investigating the antiproliferation effect and the induction of apoptosis. Apparent DNA fragmentation and morphological changes occurred in U937 cells after incubating for 48 h to 72 h with the conditioned medium that had been prepared with 400 microg CR solids/mL. Flow cytometric analysis revealed that the percentage of apoptotic U937 cells increased in a time- and concentration-dependent manner. The upregulation of Bax expression, the downregulation of Bcl-2 and procaspase-3 expression, and the release of cytochrome c from the mitochondria in U937 cells were all observed. Reverse transcription-polymerase chain reaction detected cytokine-related mRNA expressions in human mononuclear cells incubated with CR. An increase in the concentration of CR in culturing medium downregulated granulocyte macrophage colony-stimulatory factor mRNA expression while upregulated interleuken-2 mRNA expression. All the above-mentioned evidences suggest that CR induces the apoptosis of human leukemic U937 cells via the changes in cytokine profile and protein expressions in mitochondria pathway and that CR has the potential to be used in the therapy of leukemia due to its strong apoptosis-promoting effect.

Adjuvant chemotherapy with 5-fluorouracil, doxorubicin and mitomycin-C (FAM) for 6 months after curative resection of gastric carcinoma.

AIM: This study aimed to evaluate the efficacy and safety of 5-fluorouracil (5-FU), doxorubicin and mitomycin-C (FAM) adjuvant chemotherapy in patients who had undergone curative resection of gastric carcinoma. METHODS: From Nov 1999 to Jan 2002, 291 consecutive patients with stage IB-IIIB gastric adenocarcinoma were given FAM adjuvant chemotherapy. Chemotherapy comprised intravenous 5-FU 600 mg/m(2) (days 1, 8, 29 and 36), doxorubicin 30 mg/m(2) (days 1 and 29) and mitomycin-C 10 mg/m(2) (day 1), every 8 weeks for 6 months. RESULTS: The median follow-up time was 60.6 months, 92 patients died, and 93 patients had recurrent disease. The 5-year overall survival (OS) rates were 85.9% for stage IB, 72.1% for stage II, 58.0% for stage IIIA, and 48.2% for stage IIIB (p=0.002). The 5-year relapse-free survival (RFS) rates were 85.2% for stage IB, 71.2% for stage II, 53.3% for stage IIIA, and 39.2% for stage IIIB (p<0.001). A total of 769 cycles of chemotherapy were delivered, and 15 patients experienced grade 3 or higher leukopenia. The most common grade 3 or higher non-hematologic toxicity was nausea/vomiting (11 patients), followed by stomatitis (3 patients). CONCLUSIONS: Adjuvant chemotherapy with FAM for 6 months for gastric carcinoma indicated comparable RFS and OS with an acceptable toxicity profile.

Phase II study of capecitabine and cisplatin as first-line combination therapy in patients with gastric cancer recurrent after fluoropyrimidine-based adjuvant chemotherapy.

To evaluate the efficacy and safety of capecitabine and cisplatin in patients with recurrent gastric cancer after fluoropyrimidine-based adjuvant therapy. Patients with histologically confirmed and measurable advanced gastric cancer that had relapsed after fluoropyrimidine-based adjuvant chemotherapy received oral capecitabine (1250 mg m(-2) twice daily, days 1-14) and intravenous cisplatin (60 mg m(-2) over 1 h, day 1) every 3 weeks. In total, 32 patients were enrolled, of whom 30 were evaluable for efficacy and 32 for safety. A median of 5 cycles (range 1-10) was administered. One patient achieved a complete response and eight had partial responses, giving an overall response rate of 28% (95% CI, 13-44%). The median time to progression and median overall survival were 5.8 months (95% CI, 4.1-7.5 months) and 11.2 months (95% CI, 5.5-16.9 months), respectively. Grade 3 neutropenia and thrombocytopenia were observed in 38 and 6% of patients, respectively. Grade 2/3 nonhaematological toxicities included diarrhoea (19%), stomatitis (19%) and hand-foot syndrome (31%). No grade 4 toxicity, neutropenic fever or treatment-related deaths occurred. Capecitabine in combination with cisplatin was effective and well tolerated as first-line treatment in patients with recurrent gastric cancer after fluoropyrimidine-based adjuvant chemotherapy.

A phase III randomized trial of 5-fluorouracil, doxorubicin, and mitomycin C versus 5-fluorouracil and mitomycin C versus 5-fluorouracil alone in curatively resected gastric cancer.

BACKGROUND: A phase III single-center randomized trial was performed in order to determine whether the addition of mitomycin C (MMC) and/or doxorubicin to 5-fluorouracil (5-FU) as adjuvant chemotherapy could influence survival in patients with curatively resected gastric cancer. PATIENTS AND METHODS: A total of 416 patients who had undergone curative resection for stage IB-IIIB gastric adenocarcinoma were stratified according to the stage and type of surgery, and then randomized to receive one of the three chemotherapy regimens, 5-FU alone (F) or 5-FU and MMC (FM) or 5-FU, doxorubicin and MMC (FAM) within 5 weeks after surgery. RESULTS: Of 416 patients registered, 395 (133 in F, 131 in FM and 131 in FAM) were assessable. Median follow-up duration was 91 months. Five-year overall survival rates were 67.2% for F, 67.0% for FM and 66.7% for FAM (P = 0.97). Five-year disease-free survival rates were 62.1% for F, 63.3% for FM and 62.5% for FAM (P = 0.83). Hematological toxicities were more frequent in the FM and FAM groups, whereas stomatitis was more common in the F group. CONCLUSIONS: Compared with adjuvant 5-FU alone, the addition of MMC and/or doxorubicin to 5-FU did not influence survival in patients with resected gastric cancer.

Determination of random- and blockwise-type de-esterified pectins by capillary zone electrophoresis.

Capillary zone electrophoresis (CZE) was employed to determine the correlations between migration time and degree of esterification (DE) of pectinesterase-de-esterified pectins (PDPs) and alkaline-de-esterified pectins (ADPs) using 50 mM phosphate buffer (pH 6.5) as carrier electrolyte solution and 15 KV as applied voltage. Results showed that pectins with higher DEs exhibited shorter migration times. Linear correlation (r = 0.995) between migration time and DE of ADPs was observed, whereas down-curve correlation in PDPs was observed, regardless of the capillary length used (effective length, 30 and 60 cm). In addition, PDP appeared to migrate faster than ADP with the same DE under the same experimental conditions.

Change in particle size of pectin reacted with pectinesterase isozymes from pea (Pisum sativum L.) sprout.

Four pectinesterase (PE) isozymes were isolated by CM-Sepharose CL-6B chromatography from etiolated pea (Pisum sativum L.) sprouts and then reacted with citrus pectin (degree of esterification = 68%, 30-100 kDa) to observe the change in pectin particle size using a laser particle size analyzer. After incubation of a pectin-PE mixture (pH 6.5) at 30 degrees C for 4 h, PE 1 was observed to catalyze the transacylation reaction most remarkably, increasing the particle size from approximately 50-70 to approximately 250-350 nm, followed by PE 3, PE 2, and PE 4.

Isolation of immunoglobulin from egg yolk by anionic polysaccharides.

Isolation conditions of immunoglobulin in egg yolk (IgY) were optimized by the addition of various levels of Na-alginate (Alg), lambda-carrageenan (lambda-Cg), Na-carboxymethyl cellulose (CMC), and pectin (PC) to 6-fold diluted yolk. The mixtures were then reacted at pH 4.0-6.0 for 30 min. The optimal isolation conditions of IgY for Alg, lambda-Cg, and CMC were at the 0.1% level and at pH 5.0, while those for PC were at the 0.15% level and at the same pH. The remaining lipid and remaining protein in the supernatants thus obtained was 0.5-3.8% and 10-17%, respectively, and more than 90% of lipoproteins were precipitated. The IgY recovery was determined to be 33-74% by means of single radial immunodiffusion method when IgY was isolated under the optimal conditions. PC showed the best recovery of IgY, while lambda-Cg provided the least. The interactions between polysaccharides and lipoproteins were mainly ionic bonds, hydrophobic interactions, and hydrogen bonds as determined by the addition (0-2.0 M) of NaSCN or urea to the polysaccharide-lipoprotein precipitates.

Adjuvant doxorubicin and cyclophosphamide versus cyclophosphamide, methotrexate, and 5-fluorouracil chemotherapy in premenopausal women with axillary lymph node positive breast carcinoma.

BACKGROUND: This randomized controlled trial was to determine whether a combination chemotherapy regimen that contains anthracycline (doxorubicin and cyclophosphamide [AC]) is superior to the conventional cyclophosphamide, methotrexate, and 5-fluorouracil [CMF] combination in premenopausal women with axillary lymph node positive Stage II breast carcinoma. METHODS: Premenopausal women with lymph node positive breast carcinoma were stratified according to age (younger than 35 or 35 years or older) and the number of positive axillary lymph nodes (1-3, 4-9, or >/= 10) and then randomly assigned to receive either doxorubicin 40 mg/m(2) and cyclophosphamide 600 mg/m(2) intravenously (i.v.) every 3 weeks or cyclophosphamide 100 mg/m(2) orally on Days 1 through 14, methotrexate 40 mg/m(2) and 5-fluorouracil 500 mg/m(2) i.v. on Days 1 and 8 every 4 weeks. Both arms were scheduled for six cycles. RESULTS: The median follow-up was 57 months. Eighteen of the 55 AC patients developed recurrence compared with 16 of the 69 CMF patients. The corresponding 5-year recurrence free survival rates were 64% and 78%, respectively (P = 0.12). The site of the first recurrence for AC patients was locoregional in 7%, distant in 22%, and combined in 4%. The corresponding data for the CMF arm were 4%, 16%, and 3%, respectively. Six AC patients died compared with 9 CMF patients. The corresponding 5-year survival rates were 90% and 86%, respectively (P = 0.96). More leukopenia (52%, mostly Grade 1-2) occurred in the CMF arm than in the AC arm (33%, P = 0.001), but no febrile episode was accompanied with leukopenia. CONCLUSIONS: This study showed no difference between AC and CMF with respect to both disease free and overall survival rates in premenopausal women with axillary lymph node positive breast carcinoma.

Lithospermic acid B as an antioxidant-based protector of cultured ventricular myocytes and aortic endothelial cells of rabbits.

Lithospermic acid B, an active principle found in a Chinese herbal medicine for treating various heart ailments, was recently isolated, purified and demonstrated by us to salvage the postischemic rabbit heart from reperfusion injury. In this work, we further report that lithospermate B is able to protect each of two types of rabbit cardiocytes, namely ventricular myocytes and aortic endothelial cells against necrosis inflicted by oxyradicals generated pharmacologically with xanthine oxidase and hypoxanthine. Biochemically, the lithospermate B also inhibits the reduction of cytochrome c by superoxide radical anion. Thus, our in vitro data here are in concord with our earlier in vivo finding that lithospermic acid B is most likely an effective antioxidant-based myocardial protector.

Demonstration of the myocardial salvage effect of lithospermic acid B isolated from the aqueous extract of Salvia miltiorrhiza.

Lithospermic acid B has been isolated to > 95% purity by high performance liquid chromatography from the aqueous extract of the roots of Salvia miltiorrhiza. When infused at 5.5 mumoles/kg into the post-ischemic rabbit heart, it reduced by 62 +/- 10% (n = 8) the myocardial damage found in the saline control in a rabbit ischemia-reperfusion model.

[Proliferation of lymphocytes T and B by prehispanolone LC-5504 of Leonurus hereterophyllus Sweet].

Leonurus hereterophyllus Sweet is a traditional Chinese herbal medicine used to treat menstrual disturbances in woman. A new labdane diterpene, prehispanolone LC-5504, namely 9 alpha, 13R,15,16-diepoxy-labdane-14-en-7-one, had been isolated. The authors studied the synergism of labdane diterpene, prehispanolone LC-5504 and Con A and LPS on T cells and B cells proliferation of BALB/c female mice in vitro. They showed that: (1) proliferation of T cells was demonstrated by administration of LC-5504 of various concentrations together with Con A. This proliferation was 5-8 times stronger than that observed when Con A was used alone. Such effect on T cells was not observed when LC-5504 was used alone. (2) Proliferation of B cells was not observed whether LC-5504 was used alone or together with LPS.

[Prevention and treatment of isoproterenol induced ventricular fibrillation in rats by aqueous extract of Salvia miltiorrhiza].

Acute fatal ventricular fibrillation (VF) in male Sprague-Dawley rats was induced by subcutaneous injection of isoproterenol (1 mg or 5 mg/kg body weight) to two groups of rats of different body weights (525 +/- 21 g or 387 +/- 11 g) respectively. VF occurred in all control rats resulting in 96% death with only 4% spontaneously reverted and survived. Pretreatment of animals, with or without pentobarbital anaesthesia, with an aqueous extract of Salvia miltiorrhiza (SM-H, i.p., 5 g herb/kg body weight) significantly reduced J-point displacement and VF induced by isoproterenol. Survival rate was significantly raised compared with the control (P less than 0.05). Immediate intravenous injection of SM-H (5 g herb/kg body weight) to poisoned rats which developed VF caused 71% of them to recover temporarily their sinus rhythm and significantly prolonged their survival time (P less than 0.05).

Miltirone, a central benzodiazepine receptor partial agonist from a Chinese medicinal herb Salvia miltiorrhiza.

Ten diterpene quinones, which inhibited the binding of [3H]flunitrazepam to central benzodiazepine receptors with IC50s ranging from 0.3 to 36.2 microM, were isolated from the ethereal extract of the roots of Salvia miltiorrhiza. Among these natural products, miltirone has the highest potency (IC50 = 0.3 microM). It was orally active in an animal model used to predict clinical tranquilizing effects. Unlike diazepam, miltirone behaved as a partial agonist in the central benzodiazepine receptor binding and behavioural tests. Moreover, it produced no acute muscle relaxant effect and did not induce drug dependence and withdrawal reactions after chronic administration in mice.

Structure-activity relationship of miltirone, an active central benzodiazepine receptor ligand isolated from Salvia miltiorrhiza Bunge (Danshen).

Twenty one o-quinonoid-type compounds and one coumarin-type compound related to miltirone (1) have been synthesized with the aim to identify the key structural elements involved in miltirone's interaction with the central benzodiazepine receptor. On the basis of their inhibition of [3H]flunitrazepam binding to bovine cerebral cortex membranes, it is apparent that ring A of miltirone is essential for affinity. Although increasing the size of ring A from six-membered to seven- and eight-membered is well-tolerated, the introduction of polar hydroxyl groups greatly reduces binding affinity. The presence of 1,1-dimethyl groups on ring A is, however, not essential. On the other hand, the isopropyl group on ring C appears to be critical for binding as its removal decreases affinity by more than 30-fold. It can, however, be replaced with a methyl group with minimal reduction in affinity. Finally, linking ring A and B with a -CH2CH2- bridge results in analogue 89, which is 6 times more potent than miltirone at the central benzodiazepine receptor (IC50 = 0.05 microM).

[Protective action of Salvia miltiorrhiza aqueous extract on chemically induced acute myocardial ischemia in rats].

Aqueous extract of Salvia miltiorrhiza (SM-H) can protect the acute myocardial ischemia and arrhythmia of Sprague-Dawley rats induced by isoproterenol (ISO) or BaCl2 with the following results: (1) intraperitoneal injection (i.p.) of SM-H for 3 to 5 days or intravenously (i.v.) just once significantly reduced the death rate of the animals, (2) i.v. SM-H pretreatment significantly increased the lethal dose of BaCl2 infusion, (3) i.p. SM-H significantly decreased premature ventricular contraction, ventricular fibrillation, bradycardia and mortality rate induced by bolus i.v. of BaCl2, and (4) i.p. SM-H also significantly reduced ECG J-point displacement of rats induced by ISO.

Quaternary Alkaloids of Tinospora capillipes.

Extracts of the roots of TINOSPORA CAPILLIPES Gagnep. (Menispertnaceae) afforded the quaternary protoberberine alkaloids palmatine, jatrorrhizine, columbamine, stepharanine and dehydrodiscretamine and the quaternary aporphine alkaloids menisperine and magnoflorine.