薄膜过滤法检查蜂胶牙泰喷雾剂中微生物的限度

目的建立蜂胶牙泰喷雾剂中微生物限度活菌数的测定方法。方法参考《中国药典》《美国药典》24版等文献进行试验,并对所采用的方法进行方法学验证。结果常规法对照用阳性菌金黄色葡萄球菌、枯草芽孢杆菌、白色念珠菌的回收率<70%,薄膜过滤法回收率均大于80%。结论薄膜过滤法能有效地去除蜂胶牙泰喷雾剂的抑菌活性。用该法进行微生物限度检查,可行性强,能达到检测目的。     

The counting method for ameliorating traumatic memories

The Counting Method is a technique for modulating and mastering traumatic memories in which the therapist counts out loud to 100 while the client silently remembers a traumatic event. Immediately afterward, the recollection is reported, discussed and reframed. This method is briefly described and its use within the context of ongoing therapy is explained.     

Survivin mediates self-protection through ROS/cdc25c/CDK1 signaling pathway during tumor cell apoptosis induced by high fluence low-power laser irradiation

Survivin, an important member of inhibitor-of-apoptosis (IAP) family, can be up-regulated by various pro-apoptotic stimuli, such as UV, photodynamic therapy (PDT) and cisplatin. High fluence low-power laser irradiation (HF-LPLI) is a newly discovered pro-apoptotic stimulator. The anti-apoptotic mechanism of survivin during HF-LPLI-induced apoptosis is still not investigated. Here, we report that HF-LPLI up-regulates survivin activity through reactive oxygen species (ROS)/cdc25c protein phosphatase (cdc25c)/cyclin-dependent kinase (CDK1) signaling pathway in human lung adenocarcinoma cells (ASTC-a-1). The up-regulation of survivin activity can reduce HF-LPLI-induced apoptosis, while down-regulation of the activity can promote the apoptosis. In addition, activated survivin delays mitochondrial depolarization, cytochrome c release, caspase-9 and Bax activation, all of which are typical pro-apoptotic events of cell apoptosis induced by HF-LPLI. On the basis of the present studies, we conclude that survivin can mediate self-protection during tumor cell apoptosis caused by HF-LPLI.     

Joint Inferences on Vaccine Efficacy Against Infection and Disease with Application to the First HIV Vaccine Efficacy Trial

In many clinical trials, subjects are followed for two stages of outcomes, and it is of interest to compare the incidence of each outcome between two randomized groups. The outcome of the first stage may influence the outcome of the second stage. Moreover, the relative risks of the two outcomes may be linked, with the time-dependent profile of relative risk for the second outcome functionally dependent on that of the first. For example, during exposure to HIV, virologic and host factors simultaneously impact the probability of infection and the subsequent viral trajectories, and the efficacy of a tested vaccine to prevent infection and to prevent viral failure may work in concert. We address this problem by modeling the relationship between the stage two hazard function and covariates via Cox's proportional hazards model (Cox, 1972 Cox , D. R. ( 1972 ). Regression models and life-tables (with discussion) . J. R. Statist. Soc. B 34 : 187 – 220 [CSA] [Web of Science ®] , [Google Scholar]), with the stage one log-hazard ratio θ (·) at the first event time T _l included as a covariate. With θ(·) estimated using three methods, 1) nonparametric kernel smoothing; 2) locally parametric penalized splines; and 3) fully parametric cubic linear splines, we subsequently develop inference procedures for the regression parameter in the stage two Cox model based on each of the estimator of θ(·). The inferential procedures are studied in simulations and are illustrated with application to data from the world's first preventive HIV vaccine efficacy trial.     

Autophagy contributes to the survival of CD133+ liver cancer stem cells in the hypoxic and nutrient-deprived tumor microenvironment

Liver cancer stem cells (LCSCs) can drive and maintain hepatocellular carcinoma (HCC) growth, metastasis, and recurrence. Therefore, they are potentially responsible for the poor prognosis of HCC. Oxygen and nutrient deficiencies are common characteristics of the tumor microenvironment. However, how LCSCs adapt to oxygen- and nutrient-deprived conditions is unclear. Here, we used immunofluorescent staining and flow cytometry analysis to show that CD133+ cells were significantly enriched after hypoxia and nutrient starvation (H/S) in the human HCC cell line Huh7. Sorted CD133+ cells showed higher survival, less apoptosis, and possess higher clonogenic ability under H/S compared to the CD133− population. Under H/S, electron microscopy revealed more advanced autophagic vesicles in CD133+ cells. Additionally, CD133+ cells had higher autophagy levels as measured by both RT-qPCR and Western blotting. CD133+ cells had more accumulated GFP-LC3 puncta, which can be detected by fluorescence microscopy. The autophagic inhibitor chloroquine (CQ) significantly increased apoptosis and decreased the clonogenic capacity of CD133+ cells under H/S. Pre-culturing in H/S enhanced the sphere-forming capacity of CD133+ cells. However, CQ significantly impaired this process. Therefore, autophagy is essential for LCSCs maintenance. CD133+ cells were also found to have a higher tumor-forming ability in vivo, which could be inhibited by CQ administration. Collectively, our results indicate that the involvement of autophagy in maintenance of CD133+ LCSCs under the oxygen- and nutrient-deprived conditions that are typical of the tumor microenvironment in HCC. Therefore, autophagy inhibitors may make LCSCs more sensitive to the tumor microenvironment and be useful in improving anti-cancer treatments.     

Subneurotoxic copper(II)-induced NF-κB-dependent microglial activation is associated with mitochondrial ROS

Microglia-mediated neuroinflammation and the associated neuronal damage play critical roles in the pathogenesis of neurodegenerative disorders. Evidence shows an elevated concentration of extracellular copper(II) in the brains of these disorders, which may contribute to neuronal death through direct neurotoxicity. Here we explored whether extracellular copper(II) triggers microglial activation. Primary rat microglia and murine microglial cell line BV-2 cells were cultured and treated with copper(II). The content of tumor necrosis factor-α (TNF-α) and nitric oxide in the medium was determined. Extracellular hydrogen peroxide was quantified by a fluorometric assay with Amplex Red. Mitochondrial superoxide was measured by MitoSOX oxidation. At subneurotoxic concentrations, copper(II) treatment induced a dose- and time-dependent release of TNF-α and nitric oxide from microglial cells, and caused an indirect, microglia-mediated neurotoxicity that was blocked by inhibition of TNF-α and nitric oxide production. Copper(II)-initiated microglial activation was accompanied with reduced IкB-α expression as well as phosphorylation and translocation of nuclear factor-κB (NF-κB) p65 and was blocked by NF-κB inhibitors (BAY11-7082 and SC-514). Moreover, copper(II) treatment evoked a rapid release of hydrogen peroxide from microglial cells, an effect that was not affected by NADPH oxidase inhibitors. N-acetyl-cysteine, a scavenger of reactive oxygen species (ROS), abrogated copper(II)-elicited microglial release of TNF-α and nitric oxide and subsequent neurotoxicity. Importantly, mitochondrial production of superoxide, paralleled to extracellular release of hydrogen peroxide, was induced after copper(II) stimulation. Our findings suggest that extracellular copper(II) at subneurotoxic concentrations could trigger NF-κB-dependent microglial activation and subsequent neurotoxicity. NADPH oxidase-independent, mitochondria-derived ROS may be involved in this activation.     

Basic numerical processing in left intraparietal sulcus (IPS) acalculia

We report a case of a patient, AD, who suffered from acalculia following an infarct restricted to the left IPS (intraparietal sulcus). AD showed deficits in an arithmetic battery and in basic numerical processing tasks. Compared to matched controls, AD showed a larger distance effect in numerical comparisons, an abnormal size congruity indicated by a lack of facilitation by irrelevant numerical values in physical comparisons, a deficiency in counting in comparison tasks, and a deficiency in subitizing in number naming tasks and comparison tasks. This suggests that the underlying deficit in AD is a difficulty in the perception and manipulation of quantity. Moreover, it indicates the essential role of the IPS in basic numerical processing.     

Anti-melanoma activity of Cynanchi atrati Radix is mediated by regulation of NF-kappa B activity and pro-apoptotic proteins

Ethnopharmacological relevance

Cynanchi atrati Radix has been traditionally prescribed for patients with inflammatory fever or chronic tumoral disorders. Melanoma is one of the most devastating cancer types, in which overexpression of nuclear factor kappa B (NF-κB) enables the cancer to survive without apoptosis. To identify a potential anti-melanoma candidate, we evaluated the apoptotic activity of an ethanol extract of Cynanchi atrati Radix (CAE) on melanoma and its underlying mechanisms.

Materials and methods

Sixty C57BL/6 N mice with melanoma were orally administrated CAE (100 or 200 mg/kg) or distilled water for 10 days. Survival, tumor weight and volume were monitored and measured. Intratumoral apoptotic change was measured using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. To confirm the pro-apoptotic activity of CAE (10, 50 or 100 μg/mL) compared to positive drug (10 μg/mL of IKK-2 inhibitor IV), cell proliferation, caspase-3/7 activity, flow cytometric analysis, TUNEL and DAPI staining, immunoblotting and gene expression analyses for apoptosis-associated genes were conducted using B16F10 cell line.

Results

CAE administration remarkably improved survivability with a significant reduction in tumor weight (p<0.01) and volume (p<0.01), as well as increased apoptotic bodies in melanoma tissue. The CAE treatment significantly inhibited proliferation of B16F10 cells (p<0.001), but increased caspase-3/7 activity (p<0.01 or 0.001) and apoptotic population. The CAE partially blocked nuclear translocation of NF-κB but activated the p53-associated apoptotic pathway.

Conclusion

These results indicate that the CAE has anti-melanoma potential, and the underlying mechanisms involve inhibition of the activities of NF-κB and its target proteins as well as promoting the activities of pro-apoptotic proteins.     

ABBOTT CD-3700全自动血液分析仪对低值血小板检测准确度探讨

目的探讨全自动血液分析仪对低值血小板检测的准确度。方法将ABBOTF CELL—DYN3700血液分析仪检测出的102例血小板小于50×10^9／L的血常规标本同时用血小板计数参考方法和手工计数法进行比对,使用受试者工作特征曲线（Receiver Operating Characteristic Curve,简称ROC曲线）观察血液分析仪对其检测的准确度。结果血液分析仪对102例低值血小板曲线下的面积（Area Under the Curve,简称Area）为0．893,血小板数在30×10^9／L～50×10^9／L时,Area0．846;当血小板数在10×10^9／L-30×10^9／L时,Area0．746;当血小板数在0×10^9/L-10×10^9／L,Area0．650。102例手工计数的血小板ROC Area0．759,当血小板数在30×10^9／L-50×10^9／L时,ROC Area 0．702,当血小板数在10×10^9／L～30×10^9／L时,ROC Area 0．780;当血小板数在0×10^9／L～10×10^9/L时,Area为0．565。结论全自动血液分析仪CD-3700对低值血小板（〈50×10^9／L）的检测准确度较好（Area0．893）,但随着血小板数值的降低,其准确度明显下降。当血小板数值低于10×10^9／L时,分析仪已无法保证检测准确性,而传统的手工计数准确性较差,差异较大,无法起到良好的复核作用。建议采用流式细胞术对此类标本进行复检。