A in vitro platelet aggregation bioassay was developed for the quality control of XST capsules. The in vitro anti-platelet aggregation effect in rats was observed to detect the bioactivity of XST capsules. Panax notoginseng saponins and Xuesaitong lyophilizedpowder for injection were taken as standard control substances to determine the potency. According to the results, XST capsules showeda significant inhibitory effect on thrombin-induced platelet aggregation in a dose-dependent manner. The in vitro anti-platelet activity oflyophilized powder for injection was stabler than that of Panax notoginseng saponins, and so suitable to serve as a standard control substance. The biological potency of XST capsules compared with standard control substance was detected by using parallel line assay. According to the results, the established bioassay method had a good repeatability (RSD 2.92%). The sample test results could pass thereliability test(linear deviation P > 0.05, parallel deviation P > 0.05). This bioassay method could be used as one of the complementary quality control methods for XST capsules.
Animals ; Capsules ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Male ; Panax notoginseng ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology

To determine the role of vagi in heart rate variability, conscious rabbits were employed and electrocardiogram was recorded under conditions of bilateral vagi intact, unilateral vagotomy, and bilateral vagotomy. The variability of RR intervals (RRI) was analyzed using power spectrum and approximate entropy (ApEn). The results showed that the values of high frequency power (HF) component, low frequency power (LF) component and ApEn in animals with bilateral vagi intact were the highest, but the LF/HF ratio was the lowest; unilateral vagotomy decreased ApEn, right vagotomy increased LF/HF ratio but left vagotomy did not; the LF/HF ratio increased while ApEn decreased significantly in animals with bilateral vagotomy. It is suggested that the variability of RRI is mainly regulated by the vagi and the role of right vagi is more important than that of the left. When measuring heart rate variability, the results obtained with conventional method are consistent with those with nonlinear method.
Animals ; Entropy ; Heart Rate ; physiology ; Male ; Rabbits ; Vagus Nerve ; physiology

OBJECTIVEThis study aimed to investigate the mechanism of primary cortical neuron injury induced by high concentrations of copper by observing the effect of aceticum culture medium on apoptosis of rat primary cortical neurons and expression of cleaved caspase 3, caspase 8 and caspase 9.

METHODSPrimary cortical neurons were cultured for 72 hrs and then exposed to different concentrations of aceticum culture medium (20, 40 and 80 microM). The viability of neurons was detected by the MTT method. Apoptosis was observed by Hoechst33258 and flow cytometry Annexin V/PI. Expression of caspase 3, caspase 8 and caspase 9 was measured by Western blot.

RESULTSFollowing incubation with aceticum culture medium, apoptosis of neurons was induced. Theviability of neurons was remarkably reduced and the rate of apoptosis was tremendously increased in a concentration dependent manner. Caspase 8 and caspase 9 were activated in 20 microM of copper aceticum culture medium 4 hrs after incubation and peaked at 48 hrs in various concentrations of copper aceticum culture medium, presenting with a time and concentration dependent manner. The activated caspase 3 was observed in 20 microM of copper aceticum culture medium 24 hrs after incubation, which was later than the activated caspase 8 and caspase 9. Caspase 3 expression reached a peak 48 hrs in various concentrations of copper aceticum culture medium, presenting with a time and concentration dependent manner.

CONCLUSIONSThe apoptosis of primary cortical neurons can be induced by copper. Caspase 3, caspase 8 and caspase 9 cascade reaction may involve in the apoptosis of copper induced rat primary cortical neurons.

Animals ; Apoptosis ; drug effects ; Blotting, Western ; Caspase 3 ; physiology ; Caspase 8 ; physiology ; Caspase 9 ; physiology ; Cell Survival ; drug effects ; Cells, Cultured ; Cerebral Cortex ; drug effects ; Copper ; toxicity ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo establish a simple, rapid and economical method in detecting mutations of oncogene K-ras and to investigate its mutations in colorectal cancer tissues and its relationship with clinicopathologic characteristics of colorectal carcinoma.

METHODSForty colorectal cancer tissues were tested for K-ras mutations at codon 12 and codon 13 using polymerase chain reaction (PCR) followed by direct sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) followed by sequence analysis. The other 113 colorectal cancer tissues were tested for K-ras mutations at codon 12 and codon 13 using PCR-RFLP followed by sequence analysis only. The mutation results were analyzed with the corresponding clinical pathological data.

RESULTSAmong 40 colorectal cancer cases, none of K-ras mutations at codon 12 and codon 13 was detected by PCR followed by direct sequencing. However, K-ras mutations were found in 11 cases (11/40, 27.5%) by PCR-RFLP followed by sequence analysis, including 8 cases at codon 12 and 3 cases at codon 13 respectively. Among 153 colorectal cancer cases, point mutations were detected by PCR-RFLP followed by sequence analysis in 58 cases (37.9%). Point mutations at codon 12 were found in 46 cases and 12 cases at codon 13. Mutations with the highest frequency were G→A transitions (25/58, 43.1%) at codon 12. No significant correlation was observed between mutations of K-ras and gender, invasive depth, tumor differentiation, number of invaded lymph nodes, distant metastasis and clinical stage (P > 0.05). Mutation of oncogene K-ras at codon 12 and codon 13 was closely related with age and tumor location (P < 0.05). The incidence of K-ras mutation was significantly higher in younger patients and in patients with ascending colon cancer.

CONCLUSIONSPCR-RFLP followed by sequence analysis is a rapid, simple, sensitive and low-cost method. It is a suitable technology for detecting hot-spot mutations in the K-ras oncogene. Mutation of oncogene K-ras at codon 12 and codon 13 is a common molecular event in colorectal carcinogenesis, which might be related with age and tumor location.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; genetics ; pathology ; Female ; Genes, ras ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Polymorphism, Restriction Fragment Length

OBJECTIVETo investigate suitable treatment method for contracture of inframammary scars.

METHODSNine female patients with contracture of inframammary sear hospitalized in our hospital from July 2000 to July 2007 were subjected to skin expansion around the breast. The sites of incisions were mainly located on the inframammary scars. The expanders were placed around the breast and middle chest near the sternum. On the lateral side of chest, the expander should be inserted at the site parallel to upper level of the breast. The expanders should be placed under deep fascia and superficial to the gland. At II stage of operation, the scars were excised and the subcutaneous tissues should be thoroughly loosened to assure that the soft tissue and mammary gland would be restored to its anatomical position. Expanded skin was then designed as advancement or transposition flaps to repair the defects, or effects were closed with suturing.

RESULTSBlood circulation disturbance occurred at the tip of a flap in one patient, with the size of 4.0 cm x 3.0 cm, and the resulting wound healed after skin grafting. Flaps in the other 8 patients survived, and the wounds healed satisfactorily. Nipples and mammary areola were successfully restored to the anatomical positions. Three patients were followed up for 6 months to 2 years, and the result was satisfactory.

CONCLUSIONSExpanded flap is feasible for repairing contracture of inframammary scar and with good result.

Adolescent ; Breast ; Child ; Cicatrix ; surgery ; Female ; Humans ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Tissue Expansion ; methods ; Young Adult

Objective To evaluate the use of gated SPECT in patients with hypertrophic obstructive cardiomyopathy (HOCM) and the effects of percutaneous transluminal septal myocardial ablation (PTSMA) on myocardial perfusion.Methods 99 Tcm-methoxyisobutylisonitrile (MIBI) and 18F-fluorodeoxyglucose (FDG) images were performed in 31 HOCM patients before PFSMA and in 15 patients 3-7 d after PTSMA.The images in different left ventricular segments were analysed by using scores.Results In 99Tcm-MIBI images, uptake decreased at the septal regions in 12 HOCM patients (80.0%, 12/15) after PTSMA, 18F-FDG images also showed decreased uptake at the septal regions in 5 HOCM patients (33.3%, 5/15) after PTSMA.Conclusion 99Tcm-MIBI images might be an important method to evaluate PTSMA results, and 18 F-FDG images showed important value as reference.

OBJECTIVE This study was to investigate the effects of CP- 25 on the functions of activated human B cells through regulating BAFF and TNF-alpha signaling. METHODS B cells from peripheral blood mononuclear cells (PBMCs) of normal human were isolated using magnetic cell separation (MACS) by a positive selection. B cells (107 cells·mL-1) were stimulated by BAFF (100 ng·mL-1) or TNF-alpha (100 ng·mL-1) for two hours, and then were treated with CP-25 (10-5 mol·L-1) or Rituximab (5 μg·mL-1) or Etanercept (10 μg·mL-1). B cell proliferation was detected by CCK-8. B cell subsets and BAFF receptors (BAFFR, BCMA and TACI) were analyzed by flow cytometry. The expression of TNFR1 and TNFR2 on B cells was analyzed by flow cytometry. The expression of MKK3, MKK6, P-p38, P-p65, TRAF2 and p100/52 was analyzed by Western blotting. RESULTS CP-25 inhibited B cells proliferation stimulated by BAFF or TNF- alpha. CP- 25, Rituximab and Etanercept reduced the percentage and numbers of CD19+ B cells, CD19+CD20+ B cells, CD19+CD27+ B cells and CD19+CD20+CD27+ B cells induced by BAFF or TNF-alpha. CP-25 down-regulated the high expression of BAFFR, BCMA and TACI stimulated by BAFF or TNF-alpha. CP-25, Rituximab and Etanercept down-regulated significantly the expression of TNFR1 and TNFR2 on B cell stimulated by BAFF or TNF-alpha. CP-25, Rituximab and Etanercept down-regulated the expression of MKK3, P-p38, P-p65, TRAF2 and p52 in B cells stimulated by BAFF and the expression of TRAF2 and P- p65 in B cells stimulated by TNF-alpha. CONCLUSION CP- 25 regulated moderately activated B cells function by by regulating the classical and alternative NF-κB signaling pathway mediated by BAFF and TNF-alpha-TRAF2-NF-κB signaling pathway. This study suggests that CP-25 may be a promising anti-inflammatory immune and soft regulation drug.

OBJECTIVE To investigated the regulatory effect of paeoniflorin-6'-O-benzene sulfonate (CP-25) on B cell activating factor (BAFF)/BAFF receptor-nuclear factor of kappa B (NF-κB) signaling in B cell of collagen induced-arthritis (CIA) mice. METHODS Mice CIA was induced by injection of typeⅡcollagen (CⅡ). The arthritis index (AI) and swollen joint count (SJC) were assessed, and histopathology of spleen and joints were observed. The percentage of B cells subsets, BAFF receptor expressions were analyzed by flow cytometry. BAFF and immunoglobulin (Ig) levels were measured by protein antibody array. The expressions of TRAF2, MKK3, MKK6, p-P38, and p-NF-κB65 in NF-κB signaling mediated by BAFF were analyzed by western blot. RESULTS CP-25 decreased AI and SJC, restored abnormal weights, reduced thymus index and spleen index, inhibited T/B cells proliferation, alleviated the histopathology of spleen and joints in CIA mice. CP-25 also reduced high levels of serum BAFF and immunoglobulin, decreased CD19+B cells, CD19+CD27+B cells, and CD19-CD27+CD138+ plasma cells, inhibited BAFFR and TACI expressions, decreased the expressions of TRAF2, MKK3, MKK6, p-P38, and p-NF-κB65. Compared with biological agents etanercept and rituximab, CP-25 restored high T cells proliferation and percentages of B subsets to normal level, and recovered the high levels of IgA, IgD, IgG1, IgG2a and high expressions molecules in NF- κB signaling to normal levels. The action intensity of rituximab and etanercept was more strong than CP- 25. The inhibitor effects of rituximab and etanercept on AI and SJC, thymus index, proliferation of T cells and B cells subsets were strong, and down-regulated the indexes to under normal levels. CONCLUSION CP-25 might be a promising anti- inflammatory immune and regulation drug, which alleviated CIA and regulated the functions of B cells through BAFF/BAFF receptor-NF-κB signaling.

OBJECTIVETo evaluate the association between serum level of leptin and leptin receptor gene (LEPR) polymorphism and patients with breast cancer.

METHODSLEPR G1n223Arg polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism in 94 patients with breast cancer and 128 healthy controls. The level of leptin were analyzed by enzyme linked immunosorbent assay.

RESULTSIn univariate regression analyses, we found serum level of leptin and LEPR Gin223Arg genotype polymorphism were significantly higrer than those of the controls (P < 0.05-0.001, respectively). Through multivariable analyses, we found that increased risk estimates for breast cancer were among those with leptin level (OR = 1.53, 95% CI: 1.13-2.07, P = 0.006), LEPR Gin223Arg genotype (OR = 4.87, 95%CI:1.30-18.22, P = 0.019), WHR (OR = 3.68, 95% CI: 1.34-10.11, P = 0.011).

CONCLUSIONResults from this study suggested that LEPR Gln233Agr polymorphism, the elevated WHR and serum level of leptin might be correlated with increased risk of breast cancer.

Breast Neoplasms ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; Female ; Genetic Predisposition to Disease ; Humans ; Leptin ; blood ; Lipids ; blood ; Polymorphism, Genetic ; Receptors, Leptin ; genetics ; Risk

Adult ; Female ; Hepatitis B, Chronic ; complications ; physiopathology ; Humans ; Liver ; physiopathology ; Liver Function Tests ; Male ; Retrospective Studies ; Severe Acute Respiratory Syndrome ; complications ; physiopathology