Ginkgolide B Regulates CDDP Chemoresistance in Oral Cancer via the Platelet-Activating Factor Receptor Pathway.

The platelet-activating factor receptor (PAFR) is a key molecule that participates in intracellular signaling pathways, including regulating the activation of kinases. It is involved in cancer progression, but the detailed mechanism of its chemosensitivity is unknown. The purpose of the current study was to elucidate the mechanism regulating cisplatin (CDDP) sensitivity through PAFR functions in oral squamous cell carcinoma (OSCC). We first analyzed the correlation between PAFR expression and CDDP sensitivity in seven OSCC-derived cell lines based upon cell viability assays. Among them, we isolated 2 CDDP-resistant cell lines (Ca9-22 and Ho-1-N-1). In addition to conducting PAFR-knockdown (siPAFR) experiments, we found that ginkgolide B (GB), a specific inhibitor of PAFR, enhanced both CDDP chemosusceptibility and apoptosis. We next evaluated the downstream signaling pathway of PAFR in siPAFR-treated cells and GB-treated cells after CDDP treatment. In both cases, we observed decreased phosphorylation of ERK and Akt and increased expression of cleaved caspase-3. These results suggest that PAFR is a therapeutic target for modulating CDDP sensitivity in OSCC cells. Thus, GB may be a novel drug that could enhance combination chemotherapy with CDDP for OSCC patients.

Tumor Suppressive Circular RNA-102450: Development of a Novel Diagnostic Procedure for Lymph Node Metastasis from Oral Cancer.

Circular RNAs (circRNAs), which form as covalently closed loop structures, have several biological functions such as regulation of cellular behavior by adsorbing microRNAs. However, there is limited information of circRNAs in oral squamous cell carcinoma (OSCC). Here, we aimed to elucidate the roles of aberrantly expressed circRNAs in OSCC. CircRNA microarray showed that circRNA-102450 was down-regulated in OSCC cells. Clinical validation of circRNA-102450 was performed using highly sensitive droplet digital PCR in preoperative liquid biopsy samples from 30 OSCC patients. Interestingly, none of 16 studied patients with high circRNA-102450 had regional lymph node metastasis (RLNM), whereas 4 of 14 studied patients (28.5%) with low expression had pathologically proven RLNM. Overexpressed circRNA-102450 significantly inhibited the tumor metastatic properties of cell proliferation, migration, and invasion. Furthermore, circRNA-102450 directly bound to, and consequently down-regulated, miR-1178 in OSCC cells. Taken together, circRNA-102450 has a tumor suppressive effect via the circRNA-102450/miR-1178 axis and may be a novel potential marker of RLNM in OSCC patients.

Extremely enhanced N2+ lasing in a filamentary plasma grating in ambient air.

Cavity-free air lasing offers a promising route towards the realization of atmospheric lasers for various applications such as remote sensing and standoff spectroscopy; however, achieving efficient generation and control of air lasing in ambient air is still a challenge. Here we show the experimental realization of a giant lasing enhancement by three to four orders of magnitude in ambient air for the self-seeded N2+ lasing at 428 nm, assigned to the B2Σu+(ν'=0) and X2Σg+(ν''=1) emission, by modulating the spatiotemporal overlap of ultrashort near-infrared control-pump pulses in a filamentary plasma grating; meanwhile, the spontaneous emission from the same transition is only enhanced by three to four times. We find that this enhancement is sensitive to the relative polarization and interference time of the two pulses, and reveal that the formation of the plasma grating induces different population variations in the B2Σu+(ν'=0) and X2Σg+(ν''=1) levels, resulting in an enormous population inversion between the two levels, thereby a higher gain for the giant enhancement of N2+ lasing in ambient air.

Engineered exosomes delivering specific tumor-suppressive RNAi attenuate oral cancer progression.

Exosomes are involved in a wide range of biological processes in human cells. Considerable evidence suggests that engineered exosomes (eExosomes) containing therapeutic agents can attenuate the oncogenic activity of human cancer cells. Despite its biomedical relevance, no information has been available for oral squamous cell carcinoma (OSCC), and therefore the development of specific OSCC-targeting eExosomes (octExosomes) is urgently needed. We demonstrated that exosomes from normal fibroblasts transfected with Epstein-Barr Virus Induced-3 (EBI3) cDNA were electroporated with siRNA of lymphocyte cytoplasmic protein 1 (LCP1), as octExosomes, and a series of experiments were performed to evaluate the loading specificity/effectiveness and their anti-oral cancer cell activities after administration of octExosomes. These experiments revealed that octExosomes were stable, effective for transferring siLCP1 into OSCC cells and LCP1 was downregulated in OSCC cells with octExosomes as compared with their counterparts, leading to a significant tumor-suppressive effect in vitro and in vivo. Here we report the development of a new valuable tool for inhibiting tumor cells. By engineering exosomes, siLCP1 was transferred to specifically suppress oncogenic activity of OSCC cells. Inhibition of other types of human malignant cells merits further study.

Giant Enhancement of Air Lasing by Complete Population Inversion in N_{2}

A fine manipulation of population transfer among molecular quantum levels is a key technology for control of molecular processes. When a light field intensity is increased to the TW-PW cm^{-2} level, it becomes possible to transfer a population to specific excited levels through nonlinear light-molecule interaction, but it has been a challenge to control the extent of the population transfer. We deplete the population in the X^{2}Σ_{g}^{+}(v=0) state of N_{2}^{+} almost completely by focusing a dual-color (800 nm and 1.6 µm) intense femtosecond laser pulse in a nitrogen gas, and make the intensity of N_{2}^{+} lasing at 391 nm enhanced by 5-6 orders of magnitude. By solving a time-dependent Schrödinger equation describing the population transfer among the three lowest electronic states of N_{2}^{+}, we reveal that the X^{2}Σ_{g}^{+}(v=0) population is depleted by the vibrational Raman excitation followed by the electronic excitation A^{2}Π_{u}(v=2,3,4)←X^{2}Σ_{g}^{+}(v=1)←X^{2}Σ_{g}^{+}(v=0), resulting in the excessive population inversion between the B^{2}Σ_{u}^{+}(v=0) and X^{2}Σ_{g}^{+}(v=0) states. Our results offer a promising route to efficient population transfer among vibrational and electronic levels of molecules by a precisely designed intense laser field.

Absolute carrier-envelope-phase dependences of single and double ionization of methanol in a near-IR few-cycle laser field.

We investigate the carrier-envelope phase (CEP) dependences of the single and double ionization processes of methanol (CH3OH) in an intense near-IR few-cycle laser field (2.1 × 1014 W/cm2) by the asymmetry in the ejection direction of CH3 + for the non-hydrogen migration channels and CH2 + for the hydrogen migration channels created through the C-O bond breaking after the ionization. Based on the absolute CEP values at the laser-molecule interaction point, calibrated by the method using intense few-cycle circularly polarized laser pulses [Fukahori et al., Phys. Rev. A 95, 053410-1-053410-14 (2017)], we confirm that methanol cations are produced by tunnel ionization and methanol dications are produced by the recollisional double ionization. We obtain the phase offset for the double ionization accompanying no hydrogen migration to be 1.85π as the absolute CEP at which the extent of the asymmetry becomes maximum. We interpret the phase shift of 0.85π from the phase offset of 1.0π for the tunnel ionization, estimated by a tunnel ionization model incorporating the chemical bond asymmetry, as the corresponding time delay associated with the electron recollisional ionization. The positive phase shift of 0.13π for the single ionization in the non-hydrogen migration channel is interpreted as the additional time (165 as) with which a methanol cation can be excited electronically prior to the decomposition. The additional phase shift of 0.22π for the single ionization in the hydrogen migration channel is interpreted as the additional time (280 as) required for a methanol cation to be excited electronically leading to the hydrogen migration prior to the decomposition.

Rotational, Vibrational, and Electronic Modulations in N_{2}

We investigate lasing of a N_{2} gas induced by intense few-cycle near-IR laser pulses. By the pump-probe measurements, we reveal that the intensity of the B^{2}Σ_{u}^{+}-X^{2}Σ_{g}^{+} lasing emission of N_{2}^{+} oscillates at high (0.3-0.5 PHz), medium (50-75 THz), and low (∼3 THz) frequencies, corresponding to the energy separations between the rovibrational levels of the A^{2}Π_{u} and X^{2}Σ_{g}^{+} states. By solving the time-dependent Schrödinger equation, we reproduce the oscillations in the three different frequency ranges and show that the coherent population transfer among the three electronic states of N_{2}^{+} creates the population inversion between the B^{2}Σ_{u}^{+} and X^{2}Σ_{g}^{+} states, resulting in the lasing at 391 nm.

Significant Enhancement of N_{2}

We show that the intensity of self-seeded N_{2}^{+} lasing at 391 nm, assigned to the B^{2}Σ_{u}^{+}(v^{'}=0)→X^{2}Σ_{g}^{+}(v^{''}=0) emission, is enhanced by 2 orders of magnitude by modulating in time the polarization of an intense ultrashort near-IR (40 fs, 800 nm) laser pulse with which N_{2} is irradiated. We find that this dramatic enhancement of the 391 nm lasing is sensitive to the temporal variation of the polarization state within the laser pulse while the intensity of the spontaneous fluorescence emission at 391 nm is kept constant when the polarization state varies. We conclude that a postionization multiple-state coupling, through which the population can be transferred from the X^{2}Σ_{g}^{+} state of N_{2}^{+} to the first electronically excited A^{2}Π_{u} state, leads to the depletion of the population in the X^{2}Σ_{g}^{+} state, and consequently, to the population inversion between the X^{2}Σ_{g}^{+} state and the B^{2}Σ_{u}^{+} state.

Eicosapantaenoic acid treatment based on the EPA/AA ratio in patients with coronary artery disease: follow-up data from the Tochigi Ryomo EPA/AA Trial in Coronary Artery Disease (TREAT-CAD) study.

Eicosapentaenoic acid (EPA) administration has been reported to decrease the incidence of cardiovascular events, and the serum EPA/arachidonic acid (AA) ratio has been identified as a potential new risk marker for coronary artery disease (CAD). The present study aimed to investigate the value of EPA treatment based on the EPA/AA ratio at baseline. We retrospectively analyzed clinical outcome data from 149 CAD patients with a baseline EPA/AA ratio ≤ 0.4 who had received purified EPA (EPA group) or not (no EPA group) and CAD patients with an EPA/AA ratio > 0.4 who had not received EPA (control group). The baseline EPA/AA ratios were similar in the EPA and no EPA groups and were significantly lower than those in the control group (P < 0.0001). The EPA/AA ratio significantly increased in the EPA group (P < 0.0001) and the no EPA group (P < 0.001) but not in the control group. The cumulative incidence of cardiovascular death tended to be lower in the EPA group (log-rank test: P = 0.07). Receiver operating characteristic curve analysis demonstrated that the cut-off value of the target EPA/AA ratio after EPA treatment for all-cause death was 1.23 (AUC = 0.85, P = 0.016). These results suggest that EPA treatment may improve the long-term prognosis in CAD patients with an EPA/AA ratio ≤ 0.4 and that an EPA/AA ratio > 1.2 may be an appropriate EPA treatment target value to reduce mortality.

Strong-Field Fourier Transform Vibrational Spectroscopy of D_{2}

The photoionization of D_{2} and dissociation of the resultant D_{2}^{+} are monitored by pump-probe measurements using intense near-infrared few-cycle laser pulses. The yields of D_{2}^{+} and D^{+} recorded up to the pump-probe delay time of 527 ps exhibit oscillatory structures reflecting the motion of the created vibrational wave packet of D_{2}^{+}, and the Fourier transform of the data in time domain reveals the vibrational level separations with uncertainties of 0.0002-0.0097 cm^{-1}, showing a potential application of the strong-field pump-probe measurements to high-resolution spectroscopy of molecular ions.

Screening of primary aldosteronism by clinical features and daily laboratory tests: combination of urine pH, sex, and serum K.

OBJECTIVE: To develop and validate a scoring system for selection of patients who should proceed to endocrinologic examinations of primary aldosteronism in newly diagnosed hypertensive patients. METHODS: A multivariate logistic regression analysis for primary aldosteronism was undertaken by use of seven possible primary aldosteronism markers, age less than 40 years, female sex, moderate-to-severe hypertension, hypokalemia, serum Na minus Cl at least 40 mmol/l, serum uric acid 237.92 µmol/l or less (4.0 mg/dl), and urine pH (U-pH) at least 7.0, in consecutive outpatients newly diagnosed with hypertension. The diagnostic criteria of primary aldosteronism were plasma aldosterone concentration-to-plasma renin activity ratio [ARR, (ng/dl)/(ng/ml per h)] at least 20 and at least one positive result in four types of challenge tests. RESULTS: Of 130 patients, 24 were diagnosed with primary aldosteronism. The area under the receiver operating characteristic curve (AUC) for a logistic model incorporating all possible primary aldosteronism markers was 0.73 [95% confidence interval (CI): 0.61-0.85]. Removing high U-pH, female sex, and hypokalemia from the full model decreased the AUC by 0.059, 0.035, and 0.011, respectively. We devised pH of urine, female sex, low serum K (PFK) score, in which one point each was assigned to high U-pH, female sex, and hypokalemia. The prevalences of primary aldosteronism in patients with 0, 1, 2, and 3 points were 11, 14, 42, and 60%, respectively. In external validation datasets (n = 106), AUC of PFK score was significantly higher than that of hypokalemia alone (0.73, 95% CI: 0.63-0.83 vs. 0.53, 95% CI: 0.44-0.63, P < 0.01). CONCLUSION: PFK score may be a better parameter than hypokalemia alone for identifying patients with a high probability of having primary aldosteronism.

Effect of a novel calcium channel blocker on abnormal nocturnal blood pressure in hypertensive patients.

The authors examined the effect of cilnidipine, a unique L/N-type calcium channel blocker, on abnormal nocturnal blood pressure (BP) dipping in Japanese hypertensive patients in the real world. The Ambulatory Blood Pressure Control and Home Blood Pressure (Morning and Evening) Lowering by N-Channel Blocker Cilnidipine (ACHIEVE-ONE), a large-scale clinical study, was designed to evaluate the effects of cilnidipine in daily medical practice. Among the study, 24-hour ambulatory BP data were obtained from 615 patients and classified according to their nocturnal dipping status as extreme dippers, dippers, nondippers, or risers. A 12-week treatment with cilnidipine significantly reduced 24-hour BP in all groups (P<.001). Changes in nocturnal systolic BP (SBP) from baseline were -17.9 mm Hg from 154.6 mm Hg in risers and -11.9 mm Hg from 142.1 mm Hg, -6.6 mm Hg from 128.5 mm Hg, and 0.1 mm Hg from 115.8 mm Hg in nondippers, dippers, and extreme dippers, respectively. Changes from baseline in nocturnal SBP reduction rate were 8.2% in risers (P<.001) but -7.0% in extreme dippers (P<.001), while no change was observed in the nighttime SBP reduction rate for the total patients (-0.2%±9.6%, P=.617). Cilnidipine partially, but significantly, restored abnormal nocturnal dipping status toward a normal dipping pattern in hypertensive patients.

[Effectiveness of network meeting system following practical pharmacy training].

We established a network meeting system program and used it to review a prior pharmaceutical practice training session. Pharmacists at Tokai University Hachioji Hospital gave lectures about dispensing and other tasks performed by clinical pharmacists to third-year undergraduate students at Tokyo University of Pharmacy and Life Sciences. After the lectures, discussions were held using the network meeting system, after which a questionnaire was completed by the students. The questionnaire was answered by 530 students, of whom approximately 90% expressed interest in the program, 80% noted approval of the media used in the system, and 94% thought that the program was useful. Thus, we concluded that the students were motivated by the program to remember what they had learned in the lectures. We also found that the quality of data communication had an effect on the interests and motivation of the students. Based on their evaluation of the media, it was considered that improvements in communication regarding the system were necessary, though the evaluation of the utility of the program was not influenced by the quality of data communication. As a result, we concluded that our network meeting system program was useful to review prior learning of pharmaceutical practice.

[Subject making and evaluation method for objective structured clinical examination (OSCE) in pharmaceutical practice].

The aim of this study was to discuss subject selection and the evaluation method for an objective structured clinical examination (OSCE) in pharmaceutical sciences. We designed the OSCE to assess pharmaceutical students' clinical ability. In this trial, there are three stations (ST1 in circle: dispensing powdered medicine, ST2 in circle: patient reception, ST3 in circle: drug counseling), and 25 students and six instructors participated. Each student took an examination at two stations, and evaluated other students at the other station. Each instructor evaluated the student at two stations. Before the evaluation, we developed a checklist that contained the items "evaluation of the quantity" of the action, "evaluation of the quality" of the skill and the attitude, and "overall evaluation" to ensure the standardization of scoring. After the OSCE trial, we calculated and analyzed each examinee's evaluation score. In ST1 in circle and ST3 in circle, the average time for performance exceeded the time limit (5 min). There was no significant relationship between the each examinee's evaluation score and the time at all stations. The evaluation point "evaluation of the quantity" did not differ among evaluators, but a difference was seen in the "evaluation of the quality." In addition, in the quantitative evaluation, there was a difference in the evaluation of the item for which the evaluator's judgment was necessary. Instructors' evaluations were more severer than students'. In the "overall evaluation," there was no significant relation between the quantitative evaluation score and the score of the overall impression. However, there was a significant relationship with the qualitative evaluation. From these results, for assignment making, it is necessary for examinees to finish the work within the time limit, and that the evaluation not affect the performance time. Additionally, it is necessary to standardize the evaluation to reduce differences among evaluators, who should be trained. Moreover, it was suggested that the selection of an appropriate evaluation system for each evaluation item is important in OSCE stations.