Objective: To establish a quantitative analysis of multi-components by single marker (QAMS) for the simultaneous determination of six alkaloids in crude and processed Coptidis Rhizoma. Methods: An HPLC method was established to determine the six alkaloids (jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, palmatine hydrochloride, and berberine hydrochloride) by the external standard method (ESM). With this HPLC method, the berberine hydrochloride was used as the internal standard (IS) to determine five relative correction factors (RCFs) of the five other alkaloids, and their contents in all samples were calculated by their RCFs at the same time. Compared with the content results determined by the ESM and QAMS, the feasibility and accuracy of QAMS method were verified. Results: Within a certain range, the RCFs of jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, and palmatine hydrochloride to berberine hydrochloride were 1.131, 0.999, 1.011, 1.076, and 1.025, respectively, with the good repeatability in different experimental conditions. There was no significant difference between the QAMS method and ESM method. Conclusion: The QAMS method is feasible and accurate for the simultaneous determination of the six alkaloids in crude and processed Coptidis Rhizoma.

OBJECTIVETo assess the response of neoadjuvant chemotherapy and its influencing factors in the breast cancer patients.

METHODS171 patients with stage II or operable stage III breast cancers were treated with neoadjuvant chemotherapy before surgery between January 2004 and May 2005. Of these, 160 received and completed > or =3 cycles of neoadjuvant chemotherapy, 11 received only 2 cycles. The regimens of neoadjuvant chemotherapy were: CEF (CTX, Epirubicin, 5-Fu); NE (Navelbine, Epirubicin); TEC (Taxotere, Epirubicin, CTX). Response of neoadjuvant chemotherapy was evaluated in all patients by palpation, ultrasonography and pathological methods.

RESULTSComplete response rate and clinical objective response rate determined by clinical palpation (cCR, cOR), ultrasonography (sCR, sOR) and pathology (pCR) was 18.7% and 88.3%; 4.1% and 74.9%; 15.2%, respectively. The correspondence rate of the pCR with cCR and sCR was 43.8% and 42.9%, respectively. It was showed by univariate analysis that patient whose tumor was < or =3 cm in diameter, or ER negative or grade 3 were more likely to achieve a pCR than those whose tumor was >3 cm, or ER positive or grade 1. Logistic regression analysis showed that only tumor size was the significant predictive factor for response to neoadjuvant chemotherapy in patients with primary breast cancer.

CONCLUSIONPatient with small, or ER negative or grade 3 tumor may have better pathological response to neoadjuvant chemotherapy, particularly, the tumor size is more predictive of pCR. Palpation or ultrasonography may have a tendency either to under- or to overestimate pCR. Breast neoplasms/drug therapy;

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast Neoplasms ; drug therapy ; pathology ; surgery ; Carcinoma, Ductal, Breast ; drug therapy ; pathology ; surgery ; Cyclophosphamide ; administration & dosage ; Epirubicin ; administration & dosage ; Female ; Fluorouracil ; administration & dosage ; Humans ; Logistic Models ; Mastectomy ; methods ; Middle Aged ; Neoadjuvant Therapy ; Neoplasm Staging ; Receptor, Epidermal Growth Factor ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Remission Induction ; Taxoids ; administration & dosage ; Vinblastine ; administration & dosage ; analogs & derivatives

BACKGROUNDCathespin-B (cath-B) is an important proteolytic enzyme involved in the disease course of invasion in many types of cancer. Cath-B expression in subcutaneous heteroplastic pancreatic carcinoma in nude mice has not been studied. We investigated the role of cath-B in a model of heteroplastic pancreatic carcinoma in BALB/c nude mice.

METHODSThirty-two six-week-old female BALB/c nude mice were equally divided into four groups. PANC-1 cells were inoculated subcutaneously in the left axillary region. Besides volume, weight of subcutaneous tumor, and change in body weight, cath-B expression in each group was measured by immunohistochemical staining, PCR and Western blotting. Its relationship to microvessel density (MVD), CD44v6, and placenta growth factor (PLGF) was also examined. CA-074Me, a specific inhibitor of cath-B, was injected intraperitoneally (i.p.) at different stages of tumor growth in group B and C. Gemcitabine (GEM), was also injected (i.p.) in group D to compare anti-tumor efficacy with CA-074Me.

RESULTSExpression of cath-B at different levels was related to tumor growth, MVD, and PLGF expression. In group A (control group), cath-B expression was enhanced more than that seen in other groups. CA-074Me clearly inhibited cath-B expression and tumor growth in group B. There was no difference between group C and D with respect to anti-tumor effect.

CONCLUSIONSCath-B correlates with the growth and angiogenesis of tumors, but not with the adhesion induced by CD44v6. CA-074Me clearly inhibited cath-B expression and demonstrated an anti-neoplastic and anti-angiogenesis effect.

Animals ; Antineoplastic Agents ; therapeutic use ; Blotting, Western ; Body Weight ; Cathepsin B ; antagonists & inhibitors ; genetics ; metabolism ; physiology ; Cell Line, Tumor ; Dipeptides ; therapeutic use ; Female ; Humans ; In Vitro Techniques ; Mice ; Mice, Nude ; Pancreatic Neoplasms ; drug therapy ; metabolism ; Placenta Growth Factor ; Pregnancy Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Heterologous

Through cluster multistage sampling,a resident group of 6 412 subjects with hypertension in the rural area of Liaoning province were recruited.According to IDF and NCEP-ATPm criteria the age-standardized prevalences of metabolic syndrome were 36.2%and 25.9%respectively.The prevalence of metabolic syndrome decreased with advancing age,but increased with rising of blood pressure.

Objective This study observed the association between body mass index(BMI),waist circumference(WC)and blood pressure level in ratal residents from west Liarming province.Methotis This epidemiological study using stratified cluster random sampling was conducted from 2004 to 2006 in Fuxin County,Liaoning Province,43 692 rural residents(21 680 males)aged 35-74 years old[(49.8 ±10.2)years ] were surveyed.Database was established with the help of Epidata 3.1 software.Results In total 43 692 persons were surveyed,including 21 680 male(49.6%)and 22 012 female(50.4%).The average BMI and WC was(23.31±3.08)kg/m2and(80.87 ±9.40)cm,respectively.No matter male or female,SBP started from 20 kg/m2,increased with the increase of BMI;DBP increased gradually with the increase of BMI:the prevalence of hypertension were significant differences among different BMI groups (P<0.001).Multiple logistic regression show that in male,using the group with BMI,<18 kg/m2 as control,28 -30 kg/m2 group OR and 95% CI was 6. 285(4. 612 -8. 566) ; in female, when BMI >22 kg/m2 OR increased with the increasing of BMI. In male and female, both SBP and DBP, also the prevalence rate of hypertension increased gradually with the increase of WC (P <0. 001 ). No matter in male or female,when BMI < 24 kg/m2, and WC male < 85 cm, female WC < 80 cm, the average blood pressure levels and prevalence of hypertension are the lowest; after adjusting for age and other risk factors, the prevalence rate of overweight and obesity for male with hypertension OR are 1.704 ( 1. 592 - 1. 825) and 3. 710 (3. 148 -4. 371 ), respectively, for female is 1. 527 ( 1. 428 - 1. 632 ) and 3.014 ( 2. 668 - 3. 405 ), respectively.When the WC is higher than the standard, male and female hypertension risk OR and 95% CI are 1. 231( 1. 153 - 1. 314) and 1. 353 ( 1. 269 - 1. 442), respectively. Conclusion Both BMI and WC are risk factors of hypertension.

Objective To evaluate the therapeutic effects of combination administration of hydrochlorothiazide and nitrendipine at low dosage in the treatment of rural hypertension patients.Methods By the method of cluster random sampling,5292 primary hypertension patients from Fuxin,Liaoning Province were divided into health education group(control group)and drug intervention group in June 2006.The drug intervention group were treated with hydrochlorothiazide,nitrendipine and captopril by stepwise approach and we observe the antihypertensive effect of drug and the effect on the onset of stroke.Results The average follow-up time was 15 months.At last,308 patients were lost to follow-up(the lost follow-up rate was 5.8 percent).The 4984 in cohort,including 2530 of intervention group and 2454 of control group,had examination of all indicators.Through health education and drug intervention,the average blood pressure in drug intervention group decreased by 16.1/9.4 mm Hg(1 mm Hg=0.133 kPa)while the average blood pressure in control group decreased by 6.7/3.5 mm Hg.The control rate of blood pressure in drug intervention group was higher than control group(33.1%vs.15.1%,P＜0.001).Through drug intervention,the morbidity risk of nonfatal stroke in drug intervention group decreased by 57.3%compared to control group,the total morbidity risk of stroke decreased by 59.4%.The results had significant statistical difference.And,the morbidity of severe hypopotassaemia(K~+＜3.0 mmol/L)and diabetes mellitus had no significant statistical diffefence between two groups.Conclusions The low-cost antihypertensive program based on thiazide had good antihypertensive effect,high safety and good cost-effect ratio.The program could be used in rural areas of China.

To observe the expression of cyclin D1, hTERT, and telomerase activity in MNC, HL-60, HL-60A and to explore their effects on leukemogenesis and drug-resistance, normal human peripheral blood mononuclear cells, HL-60 cells sensitive to adriamycin and HL-60A cells resistant to adriamycin were investigated. The cell cycle was analyzed by flow cytometry, and the apoptosis was analyzed by Annexin V-FITC(+) PI staining. Expressions of cyclin D1 and hTERT were determined by real-time PCR and Western blot. Telomerase activity was detected by TRAP-ELISA. The results indicated that the percentage of MNC, HL-60 and HL-60A in S phase was (10.21 + 2.11)%, (44.93 + 3.00)%, and (51.38 + 1.10)% respectively; the percentage of apoptosis cells was (16.14 + 2.13)%, (7.53 + 0.92)%, (4.15 + 0.96)% respectively; the expression of mRNA and protein for cyclin D1 and hTERT increased; the telomerase activities of HL-60 and HL-60A were higher (p = 0.000), whereas the difference between HL-60 and HL-60A was no statistically significant (p = 0.232); positive correlation between cyclin D1, hTERT and telomerase activity had been found (p < 0.01). It is concluded that the cells of S phase increased while the apoptotic cells decreased in HL-60 and HL-60A, especially in HL-60A, which may be due to the up-regulation of cyclin D1, hTERT and telomerase activity.
Cell Cycle ; Cyclin D1 ; metabolism ; HL-60 Cells ; Humans ; Leukemia ; metabolism ; Telomerase ; metabolism

OBJECTIVETo explore the effects of chloride channels on the regulation of platelet cytoplasmic free calcium concentration ([Ca2+]i) and platelet aggregation (PAG).

METHODSFreshly separated platelets were activated by thrombin. Chloride channel blockers DIDS or NFA and calcium channel blockers SK&F96365 or nifedipine were added to study the effects on platelet [Ca2+]i and PAG by a single reagent or the combination of reagents and find out the interactions among DIDS, NFA, SK&F96365 and nifedipine.

RESULTSBoth DIDS and NFA could inhibit the thrombin (1 U/ml) induced PAG in a dose-dependent manner, whereas had little effect on resting [Ca2+]i. As compared with the control group, DIDS, SK&F96365 and Nifedipine could significantly reduce the PAG, Ca2+ release and Ca2+ influx in thrombin activated platelet (P < 0.05). The combination of DIDS and SK&F96365 had greater effects in reducing the PAG, Ca2+ release and Ca2+ influx than either reagent alone (P < 0.05). The combination of DIDS and nifedipine also had greater effect than each alone in reducing Ca2+ release (P < 0.05). The combination of NFA and SK&F96365 weakened each other's effect on Ca2+ release (P < 0.05), while NFA and nifedipine weakened each other's effects on PAG, Ca2+ release and Ca2+ influx in thrombin activated platelet (P < 0.05).

CONCLUSIONDIDS and NFA have no effect on the resting [Ca2+]i and the leak calcium influx of platelet. DIDS can inhibit the Ca2+ release, Ca2+ influx and PAG of platelet induced by thrombin, while NFA can only inhibit the Ca2+ release. The chloride channel and calcium channel blockers have interactions in affecting resting [Ca2+]i and PAG of platelet. The opening of chloride channel can influence the cellular calcium movement of platelet.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid ; pharmacology ; Adult ; Blood Platelets ; cytology ; drug effects ; metabolism ; Calcium ; metabolism ; Calcium Channel Blockers ; pharmacology ; Cells, Cultured ; Chloride Channels ; antagonists & inhibitors ; physiology ; Cytoplasm ; drug effects ; metabolism ; Drug Interactions ; Humans ; Imidazoles ; pharmacology ; Nifedipine ; pharmacology ; Niflumic Acid ; pharmacology ; Platelet Aggregation ; drug effects ; Thrombin ; pharmacology

OBJECTIVETo investigate the effect of valproic acid on the expression of P27(Kip1) and P170 and drug resistance of leukemia HL60/HT cell line and explore its possible mechanisms.

METHODSHL-60/HT cells were derived from HL-60 cells induced by harringtonine (HT) in gradient concentrations. The inhibitory effect of valproic acid on the proliferation of HL-60 and HL-60/HT cells was evaluated by MTT assay, and the P27(Kip1) expression, P170 expression and cell cycle of the cells were analyzed with flow cytometry.

RESULTSThe multidrug-resistant HL-60/HT was acquired, which showed a stable drug-resistant index with increased IC(50) of HT, VCR, DNR and Ara-c by 9.30, 5.20, 4.91 and 3.65 folds, respectively, as compared with those of HL60 cells. The expression of P27(Kip1) in HL-60/HT cells was significantly lower but P170 expression significantly higher than that of HL-60 cells and normal mononuclear cells (P<0.05). The expressions of P27(Kip1) and P170 showed no significant difference between normal mononuclear cells and HL-60 cells. The growth inhibition rate of VPA combined with Ara-C was significantly higher than that of valproic acid or Ara-C alone in HL-60/HT cells and HL-60 cells (q=1.37 and 1.51, respectively). HL-60/HT and HL-60 cells cultured in the presence of VPA resulted in a significant increase in the expression of P27(Kip1) and the G(1)-phase cells (P<0.05), but the expression of P170 underwent no significant changes (P>0.05).

CONCLUSIONHL-60/HT cells have lower P27(Kip1) expression compared with HL-60 cells. Valproic acid can inhibit the growth of HL-60/HT cells and enhance their Ara-C sensitivity possibly by increasing P27(Kip1) expression and causing cell cycle arrest in G(1) phase.

Antineoplastic Agents ; pharmacology ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; metabolism ; Cytarabine ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Glycoproteins ; genetics ; metabolism ; HL-60 Cells ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Valproic Acid ; pharmacology

This study was aimed to clarify whether valproic acid (VPA) induces apoptosis of leukemia HL-60 cell line and its possible mechanism. The effect of different concentrations and treatment time of VPA on HL-60 cell proliferation was assayed by cytotoxicity test (CCK-8 method) and fluorescence microscopy, and flow cytometry was used to detect cell apoptosis. The expressions of telomerase subunit h-tert mRNA and apoptosis-related protein as well as caspase-3 activity were detected by real time-quantitative PCR, Western blot and ELISA respectively. The results indicated that VPA inhibited proliferation of HL-60 cells and induced cell apoptosis in a dose dependent manner (r = -0.87). The expressions of anti-apoptotic protein BCL-2 and h-tert mRNA were significantly decreased while the pro-apoptotic protein BAX and caspase-3 activity increased after treatment with VPA. The apoptosis rate of HL-60 cell was negatively correlated with expression of h-tert mRNA. It is concluded that VPA can inhibit leukemia HL-60 cell proliferation and induce apoptosis. The VPA displays anti-leukemia activity possibly through reducing h-tert mRNA and BCL-2 protein expression, increasing BAX expression and activity of caspase-3.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; HL-60 Cells ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Telomerase ; metabolism ; Valproic Acid ; pharmacology ; bcl-2-Associated X Protein ; metabolism