Measurement of antiplatelet aggregation and potency of hirudo / 药学学报

The objective of this work was to develop a bioassay to quantify the antiplatelet aggregation activity of hirudo for quality evaluation and control. Antithrombin activity of hirudo extracted by high temperature decoction was determined by thrombin titration. Antiplatelet aggregation activity of hirudo was determined through pharmacodynamic experiments in vitro and in vivo using a bioassay we developed for quantifying inhibition of platelet aggregation. Methodological investigation was carried out and the titers of 12 batches of hirudo samples were determined. During the experiment, the disposal of animals is in accordance with the ethical standards of animal experiments. The results showed that the antithrombin activity of hirudo decocted at high temperature decreased significantly and almost lost its activity. Hirudo inhibited platelet aggregation and results in vivo and in vitro were consistent. These assays were employed to test 12 batches of hirudo. The results demonstrated that the biopotency of 12 batches was 113.49, 96.13, 121.22, 127.33, 83.48, 108.72, 131.41, 127.95, 76.90, 126.27, 132.89 and 573.53 U·mg-1. The method was reliable and reproducible and can be used to assess the quality of hirudo.

T cell exhaustion induced by persistent mycobacterium bovis BCG purified protein derivative stimulation could be restored by a Chinese Herbal Bushen Jianpi Recipe in mice model / 中国免疫学杂志

Objective:To evaluate effects of Bushen Jianpi Recipe ( BSJP) on restoring of T cell exhaustion induced by persistent tuberculosis antigens in mice.Methods:The C57BL/6 mice were primed with BCG(5×106 CFU)and boosted with 20μg of Mycobacterium bovis BCG Purified Protein Derivative ( PPD ) once a week and lasted for 16 times to induce T cell exhaustion that happened in late stage of tuberculosis .BSJP was used to treat the mice with immune exhaustion for 3 weeks.Flow Cytometry was used to detect the numbers of CD 4+and CD8+T cells and PD-1 expression.ELISA was used to detect IL-2 and IFN-γsecretion.Bacteria load in lung tissue following BCG challeng was used to detect the protective capability of mice models .Results: Compared with the transient antigen stimulation group,the persistent antigen stimulation group had lower level of IFN-γand IL-2 secretions(P<0.01).The numbers of CD4+and CD8+T cells decreased and the level of inhibitory receptor PD-1 got higher(P<0.01).The protective efficacy against BCG challenging decreased ( P<0.01 ) .After treatment with BSJP , the production of cytokines IFN-γand IL-2 increased ( P<0.01 );the numbers of CD4+and CD8+T cells increased and PD-1 expression level on CD4+T cells decreased(P<0.01);the protective efficacy against BCG challenge increased (P<0.01).The results showed that BSJP can reduce the PD-1 expression,improve the IL-2 and IFN-γsecretion,and increase the protection against BCG challenge .Conclusion:Bushen Jianpi Recipe can restore T cell exhaustion induced by persistent tuberculosis antigen stimulation in mice , which gives a hope to overcome T cell exhaustion in tuberculosis with Chinese herbal in clinic.

Study on Bushen Jianpi Recipe in treating bone marrow hematopoietic suppression mouse model caused by cyclophosphamide / 国际检验医学杂志

Objective To verify the effects of Bushen Jianpi Recipe(BSJP) on bone marrow hematopoietic suppression mouse model caused by cyclophosphamide .Methods The experimental C57BL/6 mice were divided into the normal control group ,bone marrow hematopoietic suppression group and BSJP treatment group ,and the bone marrow hematopoietic suppression group and BSJP treatment group were peritoneally injected with cyclophosphamide for 3 d .The BSJP group began to be given with BSJP by gavage from 4 d ,and the other two groups received water as gavage control .After 14 d treatment ,peripheral venous blood cells count ,bone marrow hematopoiesis stem cells (HSCs) count and bone marrow mononuclear cell proliferation ability were detected by using the blood cells analyzer and flow cytometry .Results Compared with the control group ,the white blood cells(WBCs) count , HSCs number and bone marrow mononuclear cell proliferation ability in the hematopoietic suppression group were decreased significantly(P＜0 .01);the platelet count ,HSCs count and bone marrow mononuclear cells proliferation ability after BSJP treatment in the BSJP treatment group were significantly higher than those in the hematopoietic suppression group (P＜0 .01) .Conclusion BSJP can alleviate the side effect of cyclophosphamide chemotherapy and increase the anti-tumor effect of chemotherapeutic drugs .

Functional MRI study of mild Alzheimer's disease using amplitude of low frequency fluctuation analysis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Previous studies have shown that the functional brain activity in the resting state is impaired in Alzheimer's disease (AD) patients. However, most studies focused on the relationship between different brain areas, rather than the amplitude or strength of the regional brain activity. The purpose of this study was to explore the functional brain changes in AD patients by measuring the amplitude of the blood oxygenation level dependent (BOLD) functional MRI (fMRI) signals.</p><p><b>METHODS</b>Twenty mild AD patients and twenty healthy elderly subjects participated in the fMRI scan. The amplitude of low frequency fluctuation (ALFF) was calculated using REST software.</p><p><b>RESULTS</b>Compared with the healthy elderly subjects, the mild AD patients showed decreased ALFF in the right posterior cingulate cortex, right ventral medial prefrontal cortex, and in the bilateral dorsal medial prefrontal cortex. No brain region with increased ALFF was found in the AD group compared with the control group.</p><p><b>CONCLUSIONS</b>The reduced activity in the posterior cingulate cortex and medial prefrontal cortex observed in the present study suggest that the functional abnormalities of those areas are at an early stage of AD. The ALFF analysis may provide a useful tool in fMRI study of AD.</p>

Inhibitory effect of 4-chlorobenzoyl berbamine on imatinib-resistant K562 cells in vitro and in vivo / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the inhibitory effect of 4-chlorobenzoyl berbamine (BBD9) on imatinib-resistant cell line K562 (K562/IR) in vitro and in vivo and explore the mechanisms.</p><p><b>METHODS</b>The IC50 of BBD9 and berbamine (BBM) was determined by MTT assay. The expressions of p210(Bcr-Abl), IKKa, cytoplasmic and nuclear NF-κBp65 were determined using Western blotting in K562/IR cells following a 48-h exposure to 0.5 µg/ml BBD9 or 8 µg/ml BBM. Flow cytometry was used to analyze the cell viability, apoptosis and necrosis; Western blotting was employed to determine the expressions of PARP, caspase-3, caspase-9 and LC3II in K562/IR cells exposed to different concentrations of BBD9 for 48 h. In nude mouse models bearing K562/IR cell xenograft, the tumor weight, tumor regression, and body weight changes of the mice were measured after treatments with 15 mg/kg and 30 mg/kg BBD9 and 100 mg/kg imatinib.</p><p><b>RESULTS</b>The IC50 of BBD9 and BBM was 0.73 µg/ml and 5.43 µg/ml, respectively. In K562/IR cell cultures, the expressions of p210(Bcr-Abl), IKKa and nuclear NF-κB p65 were all decreased following BBD9 and BBM treatments, but BBD9 produced more potent effect; cytoplasmic NF-κB p65 showed no obvious changes after the treatments. The cell apoptosis and necrosis increased with the concentrations of BBD9, which also dose-dependently increased the levels of cleaved caspase-3, csapase-9, PARP, and LC3II expression. In the tumor-bearing mouse model, BBD9 showed stronger effects than imatinib in reducing the tumor weight, promoting tumor regression, and increasing the body weight.</p><p><b>CONCLUSION</b>BBD9 can effectively inhibit the growth of K562/IR cells in vitro and in vivo by activating cell apoptosis, necrosis and autophage pathways, down-regulating expressions of p210(Bcr-Abl) and IKKa and suppressing the cytoplasm-to- nucleus translocation of NF-κBp65.</p>

Value evaluation of UF-1000i automated urine formed elements analyzer in the diagnosis of urinary tract infection / 中华检验医学杂志

Objective To evaluate the application value of UF-1000i automated urine formed elements analyzer in the diagnosis of urinary tract infection. Methods 150 urine specimens were analyzed using the UF-1000i in parallel with detection of leukocyte, yeast-like fungus, and bacteria. These detection results were collected for evaluation of urinary tract infection and scatter grams were recorded. At the same time, these samples were cultured for bacterial identification, which results were compared with that of the UF-1000i. The clinical diagnose criteria of the UTI was performed as golden standard. As compare with results obtained with UF-1000i, the sensitivity and specificity of UF-1000i for diagnosis of urinary tract infection were evaluated, and the consistency were analyzed among scatter grams, bacterial culture and final diagnosis. Results The statistical results from 146 specimens showed that the positive rate of UF-1000i was 32. 9% (48/146), the positive rate of urine culture is 28. 8% (42/146). There was no significant statistical difference found (χ2 = 1.79 ,P = 0. 18 )and Kappa test showed a considerable consistency (K = 0. 775 6). The UF-1000i detection results showed the sensitivity 76. 0% ( 38/50 ), specificity 89. 6% ( 86/96 ), positive predictive value 79. 2% ( 38/48 ) and negative predictive value 87. 8% ( 86/98 ), respectively. The distribution of coccus and bacilli obtained from the UF-1000i testing was basically in accordance with the results of bacterial culture. Conclusion The "UTI-information" of UF-1000i is very important for the diagnosis of urinary tract infections.