Evaluation of coating uniformity for the digestion-aid tablets by portable near-infrared spectroscopy.

Process analysis can effectively stabilize pharmaceutical quality and optimize the control of production process. For the sustained-release digestion-aid tablets, the coating film thickness is an important indicator to measure the quality of products. Traditional method mainly spot-checks tablets and measures with visual microscopy, which is time-consuming and laborious. This study attempted to use a portable near-infrared spectroscopy for rapid detection of a Chinese medicine tablets from production line. First, PLS regression models were established for coating film at twelve different locations of the tablet section, and the results showed that the correlation coefficients of training and validation sets were all over 0.80. Subsequently, the twelve locations were divided into six groups to further establish regressions. After chemometrics optimization, the optimal of six group models were generally better than single location models, with Rc2 and Rv2 all above 0.85, and RMSEV values all below 2.0. The mean relative error of prediction of the optimal model was 9.49%. The pharmaceutical process detection based on the portable NIR spectroscopy met the demand of managing digestion-aid tablet coating data conveniently. The proposed approach can successfully realize on-site and online pharmaceutical monitoring and has a promising practical value.

S-Nitrosocaptopril prevents cancer metastasis in vivo by creating the hostile bloodstream microenvironment against circulating tumor cells.

A perfect microenvironment facilitates the activated circulating tumor cells (CTCs) to spark the adhesion-invasion-extravasation metastatic cascade in their premetastatic niche. Platelet-CTC interaction contributes to the progression of tumor malignancy by protecting CTCs from shear stress and immunological assault, aiding CTCs entrapment in the capillary bed, enabling CTCs to successfully exit the bloodstream and enter the tissue, inducing epithelial-mesenchymal-like transition (EMT), and assisting in the establishment of metastatic foci. To prevent the cascade from sparking, we show that, the multifunctional S-nitrosocaptopril (CapNO) acts on both CTCs and platelets to interrupt platelet/CTCs interplay and adhesion to endothelium, thus inhibiting CTC-based pulmonary metastasis in vivo. The activated platelets cloak cancer HT29 cells, resulting in HT29-exhibiting platelet biomarkers CD61 and P-selectin positive. CapNO inhibits both sialyl Lewisx (Slex) expression on HT29 and ADP-induced activation of platelets through P-selectin- and GPIIb/IIIa-dependent mechanisms, confirmed by the corresponding antibody assay. CapNO inhibits platelet- or interleukin (IL)-1ß-mediated adhesion between HT29 and endothelial cells, and micrometastatic formation in the lungs of immunocompetent syngeneic mouse models. CapNO have also shown the effects of vasodilation, anticoagulation, inhibition of matrix metalloproteinase-2 (MMP2) expression on cancer cells, and inhibition of cell adhesion molecules (CAMs) expression on vascular endothelium. Due to a series of the beneficial effects of CapNO, CTCs remain exposed to the hostile bloodstream environment and are vulnerable to death induced by shear stress and immune elimination. This new discovery provides a basis for CapNO used for cancer metastatic chemoprevention, and might suggest regulation of the CTCs bloodstream microenvironment as a new avenue for cancer metastatic prevention.

Circulation patterns and seed-soil compatibility factors cooperate to cause cancer organ-specific metastasis.

Despite the recognition of the lethality of cancer metastasis and the importance of developing specific anti-metastasis therapies directed at the cancer metastatic cascade, the dynamics of cancer metastasis remains poorly understood. In this study, we examined the dynamics of circulating tumor cell (CTC) survival in the bloodstream using experimental mouse models. CTCs were arrested in the capillaries by adhesion to vascular endothelium within a few minutes after injection into the bloodstream. The loss of CTCs from the circulation followed a bi-phasic decay pattern, with the number of CTCs in the bloodstream being closely associated with the number of blood circulation cycles. The calculated in vivo Vd (apparent volume of distribution) of the CTC revealed organ specific binding of the CTCs. Moreover, confocal microscopy, in vivo fluorescence imaging in syngeneic mouse metastatic models and analysis of blood circulation patterns support the notion of organ-specific tumor metastasis. The present study suggests that organ-specific tumor metastasis is influenced by cooperation between blood circulation patterns and 'seed-soil' compatibility factors. These new findings provide further insights for optimized cancer metastatic prevention strategies such as by creating a hostile circulation microenvironment and targeting the organ-specific 'seed-soil' compatibility factors.

Sex-related pharmacokinetic differences and mechanisms of metapristone (RU486 metabolite).

Metapristone is the primary metabolite of the abortifacient mifepristone (RU486), and is being developed as a safe and effective cancer metastatic chemopreventive agent for both sexes. Here, we systematically investigated the sex-related pharmacokinetics of metapristone in both rats and dogs, and explored the related mechanisms of actions. Administration of metapristone to rats and dogs showed that plasma concentrations of metapristone (AUC, C max ) were significantly higher in female dogs and rats than in males. The sex-related differences in pharmacokinetics become more significant after ten consecutive days of oral administration. Female liver microsomes metabolized metapristone significantly slower than the male ones. The results from P450 reaction phenotyping using recombinant cDNA-expressed human CYPs in conjunction with specific CYP inhibitors suggested that CYP1A2 and CYP3A4 are the predominant CYPs involved in the metapristone metabolism, which were further confirmed by the enhanced protein levels of CYP1A2 and CYP3A4 induced by 1-week oral administration of metapristone to rats. The highest tissue concentration of metapristone was found in the liver. The study demonstrates, for the first time, the sex-related pharmacokinetics of metapristone, and reveals that activities of liver microsomal CYP1A2 and CYP3A4 as well as the renal clearance are primarily responsible for the sex-related pharmacokinetics.

The Architecture and Function of Monoclonal Antibody-Functionalized Mesoporous Silica Nanoparticles Loaded with Mifepristone: Repurposing Abortifacient for Cancer Metastatic Chemoprevention.

The circulating tumor cells (CTCs) existing in cancer survivors are considered the root cause of cancer metastasis. To prevent the devastating metastasis cascade from initiation, we hypothesize that a biodegradable nanomaterial loaded with the abortifacient mifepristone (MIF) and conjugated with the epithelial cell adhesion molecule antibody (aEpCAM) may serve as a safe and effective cancer metastatic preventive agent by targeting CTCs and preventing their adhesion-invasion to vascular intima. It is demonstrated that MIF-loaded mesoporous silica nanoparticles (MSN) coated with aEpCAM (aE-MSN-M) can specifically target and bind colorectal cancer cells in either cell medium or blood through EpCAM recognition proven by quantitative flow cytometric detection and free aEpCAM competitive assay. The specific binding results in downregulation of the captured cells and drives them into G0/G1 phase primarily attributed to the effect of aEpCAM. The functional nanoparticles significantly inhibit the heteroadhesion between cancer cells and endothelial cells, suggesting the combined inhibition effects of aEpCAM and MIF on E-selectin and ICAM-1 expression. The functionalized nanoparticles circulate in mouse blood long enough to deliver MIF and inhibit lung metastasis. The present proof-of-concept study shows that the aE-MSN-M can prevent cancer metastasis by restraining CTC activity and their adhesion-invasion to vascular intima.

Thirty-day rat toxicity study reveals reversible liver toxicity of mifepristone (RU486) and metapristone.

OBJECTIVE: Mifepristone (RU486) is an oral first-line contraceptive used by hundreds of millions of women, and recently it was tested for anticancer activity in both genders worldwide. We are developing metapristone (the N-monodemethyl RU486) as a potential metastasis chemopreventive. The present acute and 30-d subacute toxicity study aimed at examining and compared in parallel the potential toxicity of the two drugs. METHODS: The single-dose acute toxicity and 30-d subacute toxicity studies were conducted in mice and rats, respectively, by gavaging metapristone or mifepristone at various doses. Blood samples and organs were collected for blood chemistry, hematology and histology analyses. RESULTS: Oral mifepristone (3000 mg/kg) caused 30% and 40% death in female and male mice, respectively, within 15 h post-dosing. In comparison, the same dose of metapristone produced 30% acute death in males only. Thirty-day oral administration of the two drugs to rats (12.5, 50 and 200 mg/kg/day) caused reversible hepatotoxicity that only occurred at 200 mg/kg/day group, evidenced by the elevated liver enzyme activity and liver organ weight. CONCLUSION: The present study, for the first time, reveals reversible hepatotoxicity in rats caused by the 30-d consecutive administration at the high dose, and warns the potential hepatotoxicity caused by long-term administrations of high doses of mifepristone or metapristone in clinical trials but not by the acute single abortion doses.

In vitro and in vivo efficacy and safety evaluation of metapristone and mifepristone as cancer metastatic chemopreventive agents.

Malignant melanoma, the most deadly form of skin cancer, has a high propensity for metastatic spread and is notoriously chemotherapy-resistant. Metapristone is the primary metabolite of mifepristone (RU486) and shows biological activities similar to RU486. In the present study, we comprehensively investigated the efficacy of metapristone as a metastatic chemopreventive against melanoma B16F10 cells in vitro and in vivo, and evaluated the safety profile of both drugs in mice. Metapristone showed less cytostatic effect in vitro and in vivo in comparison with mifepristone. However, metapristone interfered the adhesion of B16F10 cells to fibronectin by down-regulating cellular expression of integrin α4. Chemopreventive pretreatment followed by oral administration of metapristone and mifepristone (2.5, 10, 50 mg/kg/day for 35 days) to melanoma C57BL/6 mouse model showed significant attenuation of pulmonary metastatic development. Oral administration of high doses of metapristone and mifepristone to normal mice for 35 days (25, 100, 250 mg/kg/day) resulted in a dose-dependent increase in mouse liver weight that was more severe with mifepristone than metapristone. The long-term toxicity study revealed more changes by mifepristone in counts of erythrocytes, leukocytes and platelets than by metapristone. In conclusion, metapristone may fit into a new class of cancer metastatic chemopreventive agents. It showed a safety and efficacy profile better than mifepristone.

[Detection of Adulteration in Milk Powder with Starch Near Infrared].

Three China trademarks of milk powder called Mengniu, Yili, Wandashan were taken as testing samples. Each of them mixed varied amount of starch in different gradient, which were consisted of 32 adulterated milk powder samples mixed with starch, was taken as standard samples for constructing predicted model. To those 32 samples, the reflecting spectrum characteristics in middle wave of near infrared spectrum with Near Infrared Spectrum Analyzer (Micro NIR 1700) produced by JDSU Ltd. USA were collected for five repeats in five different days. The time span was nearly two months. Firstly, we build the model used the reflecting spectrum characteristics of those samples with biomimetic pattern recognition (BPR) arithmetic to do the qualitative analysis. The analysis included the reliability of testing result and stability of the model. When we took ninety percent as the evaluation threshold of testing result of CAR (Correct Acceptance Rate) and CRR (Correct Rejection Rate), the lowest starch content of adulterate milk powder in all tested samples which the tested result were bigger than that abovementioned threshold was designated CAR threshold (CAR-T) and CRR threshold (CRR-T). CAR means the correct rate of accepting a sample which is belong to itself, CRR means correct rate of refusing to accept a sample which is not belong to itself. The results were shown that, when we constructed a model based on the near infrared spectrum data from each of three China trademark milk powders, respectively, if we constructed a model with infrared spectrum data tested in a same day, both the CAR-T and CRR-T of adulterate starch content of a sample can reach 0.1% in predicting the remainder infrared spectrum data tested within a same day. The three China trademarks of milk powder had the same result. In addition, when we ignored the trademarks, put the spectrum data of adulterate milk powder samples mixed with the same content of starch of three China trademarks milk powder together to construct a model, the CAR-T of mixed starch content of a sample may reach 0.1%, the CRR-T can reach 1%, if the model construction and predicting were performed with near infrared spectrum data tested in a same day. However, the CAR-T can just stably reach up to 5% and the CRR-T have the same result, if the model construction and predicting were crossly performed with mixed near infrared spectrum data tested in different days. Furthermore, the correct recognizing threshold mixed starch of a sample can stably reach up to 1% and the CAR-T can reach 5%, if the model construction was based on near infrared spectrum data combined the previous four days to predict the output of the another day. On the other hand, we also engaged quantitative analysis to the starch content in milk power with two kinds of arithmetic (PLSR, LS-SVR). In contrast with the testing outputs, the reliability of both the CAR-T and CRR-T in qualitative analysis was further validated.

Aspirin, lysine, mifepristone and doxycycline combined can effectively and safely prevent and treat cancer metastasis: prevent seeds from gemmating on soil.

Recent scientific advances have increased our understanding of the cancer metastatic complexities and provided further impetus for new combination therapies to prevent cancer metastasis. Here, we demonstrated that a combination (HAMPT) of aspirin, lysine, mifepristone and doxycycline can effectively and safely prevent cancer metastasis. The pharmaceutically-formulated HAMPT inhibited adhesion of cancer cells to either endothelial cells or extracellular matrix via down-regulating cell adhesion molecules ICAM-1 and α4-integrin. HAMPT inhibited the cloak effect by activated platelets on cancer cells, thereby interfering adhesion and invasion of cancer cells to the underlying stroma. At the effective concentration, HAMPT induced cancer cells into dormancy with minor inhibition on cell viability. Four-day pretreatment followed by 30-day oral administration of HAMPT (33.5-134 mg/kg) to the mice inoculated with cancer cells produced significant inhibition on cancer metastasis dose-dependently without marked side effects. Fifty-day rat toxicity study with HAMPT at doses (335-1340 mg/kg) 20-fold higher than its therapeutic dose produced no significant toxicity. Interestingly, the acute toxic death could not be reached at the maximum administrable dose (5 g/kg). This proof-of-concept study provides the first conceptual evidence that cancer metastasis can be controlled by using affordable old drugs to restrain circulating tumor cells from gemmating on the metastatic soil without the need for cytotoxicity.

Multivalent conjugation of antibody to dendrimers for the enhanced capture and regulation on colon cancer cells.

Circulation tumor cells (CTCs) in the bloodstream of early-stage cancer patients carry the important information about valuable biomarkers and biological properties of primary tumor. However, detection and capture of CTCs are challenging owing to their low concentrations. Traditional technologies have the limited detection sensitivity and the low capture efficiency. We, herein, report an effective approach to specifically bind and capture colon cancer HT29 cells by using multiple Sialyl Lewis X antibodies (aSlex)-conjugated PAMAM dendrimers. The conjugation was characterized by using atom force microscope, UV and fluorescence measurements. The capturing and regulating HT29 cells by the aSlex-coated dendrimer conjugate were analyzed by microscopy and flow cytometry. The results indicated that the conjugate showed the enhanced capture of HT29 cells in a concentration-dependent manner and the maximum capture efficiency of 77.88% was obtained within 1 h-exposure. G6-5aSlex-FITC conjugate showed capture efficiency better than FITC-G6-COOH-5aSlex conjugate. G6-5aSlex-FITC conjugate could specifically capture HT29 cells even when the target HT29 cells were diluted with the interfering cells (e.g., RBCs) to a low concentration. The capture resulted in a concentration-dependent restraint of the cell activity. In conclusion, the aSlex-coated dendrimer conjugate displayed the great potential in capturing and restraining colorectal CTCs in blood.

Systems pharmacology of mifepristone (RU486) reveals its 47 hub targets and network: comprehensive analysis and pharmacological focus on FAK-Src-Paxillin complex.

Mifepristone (RU486), a synthetic steroid compound used as an abortifacient drug, has received considerable attention to its anticancer activity recently. To explore the possibility of using mifepristone as a cancer metastasis chemopreventive, we performed a systems pharmacology analysis of mifepristone-related molecules in the present study. Data were collected by using Natural Language Processing (NLP) and 513 mifepristone-related genes were dug out and classified functionally using a gene ontology (GO) hierarchy, followed by KEGG pathway enrichment analysis. Potential signal pathways and targets involved in cancer were obtained by integrative network analysis. Total thirty-three proteins were involved in focal adhesion-the key signaling pathway associated with cancer metastasis. Molecular and cellular assays further demonstrated that mifepristone had the ability to prevent breast cancer cells from migration and interfere with their adhesion to endothelial cells. Moreover, mifepristone inhibited the expression of focal adhesion kinase (FAK), paxillin, and the formation of FAK/Src/Paxillin complex, which are correlated with cell adhesion and migration. This study set a good example to identify chemotherapeutic potential seamlessly from systems pharmacology to cellular pharmacology, and the revealed hub genes may be the promising targets for cancer metastasis chemoprevention.

A novel UPLC/MS/MS method for rapid determination of metapristone in rat plasma, a new cancer metastasis chemopreventive agent derived from mifepristone (RU486).

Mifepristone (RU486) is a chemical abortifacient used by hundreds of millions of women world-wide. It has recently been used in clinical trials for psychotic depression and cancer chemotherapy. Metapristone is the most predominant biological active metabolite of mifepristone, and being developed as a novel cancer metastasis chemopreventive agent based on its unique pharmacological properties. In this study, a novel rapid and sensitive method using UPLC/MS/MS was developed and validated for quantitative analysis of metapristone in plasma, which used less plasma volume and was demonstrated to be more simple and low-cost than the published methods. Metapristone in plasma was recovered by liquid-liquid extraction using 1 mL of ethyl acetate and chromatographic separation was carried on a C18 column at 35 °C, with a gradient mobile phase consisting of methanol and water containing 0.1% (v/v) formic acid at a flow rate of 0.3 mL/min. The mass spectrometric detection was carried out using a triple-quadrupole system via positive electrospray ionization. Multiple reaction monitoring was used for quantitation of m/z transitions from 416.3 to 119.9 for metapristone and from 313.1 to 109 for levonorgestrel (internal standard). Good linearity (r²> 0.9926) was achieved over a concentration range from 7.1 to 2840 ng/mL with a lower limit of quantification of 7.1 ng/mL for metapristone. The intra- and inter-day variations of the assay were 2.4-10.0% relative standard deviation with an accuracy of -5.6 to 8.6% relative error. This newly developed method was successfully applied to a pharmacokinetic study that revealed, for the first time, that there was a significant difference in pharmacokinetic profile between genders.

Synthesis, spectral characterization, and in vitro cellular activities of metapristone, a potential cancer metastatic chemopreventive agent derived from mifepristone (RU486).

Mifepristone (RU486) is marketed and used widely by women as an abortifacient, and experimentally for psychotic depression and anticancer treatments. After administration, metapristone is found to be the most predominant metabolite of mifepristone. We hypothesized that adhesion of circulating tumor cells (CTCs) to vascular endothelial bed is a crucial starting point in metastatic cascade, and that metapristone can serve as a cancer metastatic chemopreventive agent that can interrupt adhesion and invasion of CTCs to the intima of microvasculature. In the present study, we modified the synthesis procedure to produce grams of metapristone, fully characterized its spectral properties and in vitro cellular activities, including its cytostatic effects, cell cycle arrest, mitochondrial membrane potential, and apoptosis on human colorectal cancer HT-29 cells. Metapristone concentration dependently interrupted adhesion of HT-29 cells to endothelial cells. Metapristone may potentially be a useful agent to interrupt metastatic initiation.

[Reason analysis of inadaptability and its correction research on the authenticity identification model of West Lake Longjing tea based on LVF micro-NIR spectrometer].

In the present paper, the micro-NIR spectrometer with the splitter of linear variable filter was used to develop the recognition models of the West Lake Longjing tea and the ordinary flat tea of the year 2012 and 2013. The NIR spectral data of different years and different storage times were decomposed by PCA algorithm. The PLS-DA models were developed by the representative samples selected by the mathematical characteristics of PCA-scores' distribution in order to analyze the reason for the inadaptability of the models according to mathematical principles and find out the solution for its correction. Being examined by the external validation set, the adaptability of the authenticity identification model was enhanced effectively. The result of this research indicated that, for the West Lake Longjing tea and the ordinary flat tea, the correct recognition rate of the model developed by all different-year samples' NIR spectral data would be enhanced effectively. The model developed by the NIR spectral data of different storage time samples indicated that the physicochemical properties of the ordinary flat tea have changed remarkably after cryopreservation for 3 months, while the physicochemical properties of the West Lake Longjing tea are relatively stable. The model adaptabilities for different years and different storage times were studied according to the mathematical perspective of the principal component characteristics of spectral data. After the authenticity identification model of West Lake Longjing tea was developed, the prediction accuracy was enhanced effectively. This research would provide reference for not only the application of NIR spectroscopy in quality grading and safety of agricultural products, but also the enhancement of the prediction accuracy of the NIR grading models for agricultural products.

[The study of AutoFlow role in volume controlled ventilation].

OBJECTIVE: To evaluate the efficacy and safety of automatic variable flow rate (AutoFlow) for volume control ventilation through monitoring the number of ventilator alarm. METHODS: Forty-eight adult patients receiving the Drager Evita 4 ventilator with an expectation of more than 2 days duration were divided into two groups by randomly digital methods, each n=24. The patients in control group were received routinely mode with synchronized intermittent mandatory ventilation (SIMV), and the others in observation group were given SIMV and assist with AutoFlow. The midazolam and fentanyl was given to retain the Ramsay score 2-3 by continuous micro-pump. The ventilator alarm, blood gas analysis and respiratory function were recorded. RESULTS: There were no significant differences in respiratory rate (RR), tidal volume (VT), positive end-expiratory pressure (PEEP), fraction of inspired oxygen (FiO2), pH, arterial partial pressure of carbon dioxide (PaCO2), arterial partial pressure of oxygen (PaO2), oxygenation index (PaO2/FiO2), as well as sedative dose and time between two groups within 5 days of mechanical ventilation. Duration of mechanical ventilation in all patients was 164 days (3756 hours), and 78 days (1812 hours) in control group, 86 days (1944 hours) in observation group. The duration of mechanical ventilation in observation group was longer than that in control group [3 (1-15) days vs. 2 (1-28) days, P>0.05]. A total of 23 843 alarms were recorded, approximately 6 times/h, and 17 386 alarms in control group, averagely 9.6 times/h, 6457 alarms in the observation group, averagely 3.3 times/h. The number of ventilator alarm in observation group was less than that in control group (P<0.01). The number of airway pressure alarm in observation group was less than that in the control group [122 (8-1068) vs. 565 (13-1898), P<0.01]. There was no significant difference in sequential organ failure assessment (SOFA) score within 5 days between the two sets of mechanical ventilation. In the observation group ventilator-associated pneumonia (VAP) was occurred in 4 cases, and no pneumothorax happened, while in the control group there were 8 cases and 2 cases respectively. The mortality rate in intensive care unit (ICU) in observation group was lower than that in control group, but there was no statistical difference (25.0% vs. 37.5%, P>0.05). CONCLUSIONS: AutoFlow is confirmed be safe for volume control ventilation mode, and could significantly reduce the alarm of ventilator.

Sucrose as chiral selector for determining enantiomeric composition of metalaxyl by UV-vis spectroscopy and PLS regression.

This study revealed that it was possible to determine the enantiomeric composition of with multivariate regression models of spectral data obtained by ordinary UV-vis spectrophotometry of enantiomeric guest-host complexes. The total 60samples involving three concentration levels of metalaxyl as low, medium and high were prepared for spectral collecting. Four methods of modeling were subsequently proposed and compared including two common ways and two compensating ways for variations in total analyte concentration. Firstly, without normalization robust modeling was failed to achieve while employing the medium concentration levels as calibration and the other two levels as a validation. The same case occurred when full-cross validation was conducted. Besides, two enhanced methods were developed to account for the systematic variation. One of which normalized the spectra with respect to the total concentration of enantiomeric, along with spectral data, as a variable in the statistical analysis. The other one ignored variations in total concentration, relying on the specific band normalization to sort out any variations due to total concentration differences. The results clearly demonstrated that the spectra according to concentration provided the acceptable predictive ability in determining enantiomeric composition.

[Fast determination of the proportion of metalaxyl-M enantiomers by UV spectroscopy].

In the present study, beta-cyclodextrin(betaCD) was used as chiral selector to detect the proportion of chiral isomers of metalaxyl. The proportion of metalaxyl enantiomers can be detected by ultraviolet (UV)spectroscopy since the interaction between the R, S isomer of metalaxyl with beta-CD is different. The quantitative models were established by partial least squares regression (PLS) and the robust of models was evaluated by independent validation samples. The determination coefficient R2 of calibration set in the quantitative model was 0.999 0. The standard error of calibration set (SEC) and the relative standard deviation (RSD) of the model was respectively 0.006 7 and 0.89%; The correlation coefficient r of estimated value and specified value of the 6 independent validation samples was 0.998 5. The standard error of prediction (SEP) and RSD was respectively 0.008 9 and 1.17%. This method is rapid and easy to operate in practical applications.