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After ovulation, the mitochondrial enzyme CYP11A1 cleavage the cholesterol into pregnenolone for progesterone synthesis, suggesting that mitochondrial dynamics play a vital role in the female reproductive system. The changes in the mitochondria dynamics throughout the ovarian cycle have been reported in literature, but the correlation to its role in the ovarian cycle remains unclear. In this study, mitochondrial fusion promotor, M1, was used to study the impact of mitochondria dynamics in the female reproductive system. Our results showed that M1 treatment in mice can lead to the disruptions of estrous cycles in vagina smears. The decrease in serum LH was recorded in the animal. And the inhibitions of progesterone secretion and ovulations were observed in ovarian culture. Although no significant changes in mitochondrial networks were observed in the ovaries, significant up-regulation of mitochondrial respiratory complexes was revealed in M1 treatments through transcriptomic analysis. In contrast to the estrogen and steroid biosynthesis up-regulated in M1, the molecules of extracellular matrix, remodeling enzymes, and adhesion signalings were decreased. Collectively, our study provides novel targets to regulate the ovarian cycles through the mitochondria. However, more studies are still necessary to provide the functional connections between mitochondria and the female reproductive systems.

International guidelines do not recommend surgery for the first episode of primary spontaneous pneumothorax (PSP), except in cases of persistent air leak, hemopneumothorax, bilateral pneumothorax, or occupations at risk. However, these recommendations have been challenged because of a significant reduction in the recurrence rate in emerging studies. We evaluated the rationale of recommendations by systematically reviewing RCTs and observational studies by using the Grading of Recommendations, Assessment, Development, and Evaluations (GRADE) system. We searched articles in PubMed, EMBASE, and Cochrane databases up to August 15, 2020. The primary outcomes were the recurrence rate and complication rate. The secondary outcomes were hospital stay and drainage duration. Nine eligible studies with 1121 patients were retrieved and analyzed. The recurrence rate was lower in the VATS than in conservative treatment with moderate evidence (OR 0.13, 95% CI 0.09 to 0.19, P 2 = 0%). We did not find significant differences in complication rate (Peto OR 1.17, 95% CI 0.33 to 4.12, P = 0.80), hospital stay duration (MD − 0.48 days, 95% CI − 2.84 to 1.87, P = 0.69, very low evidence), and in drainage duration (MD − 3.99 days, 95% CI − 9.06 to 1.08, P = 0.12, very low evidence) between the two groups. Our results would suggest VATS treatment as a weak recommendation for patients with the first episode of PSP, based on our systematic review of the current evidence by using the GRADE system, indicating that different treatments will be appropriate for different patients and that patients’ values and preferences should be incorporated through shared decision making.Trial REGISTRY: PROSPERO; No.: CRD42020162267.

BACKGROUND Kisspeptin (KISS1) and kisspeptin receptor (KISS1R) are essential gatekeepers of the reproductive system. The functions of KISS1 and KISS1R in corpus luteal cells remain ambiguous. The objective was to observe normal physiologic functions of corpus luteal cells in vivo and clarify the functions of KISS1 in vitro. METHODS We conducted an in vivo observation of cellular patterns as well as the levels of steroidogenic enzymes and KISS1/KISS1R in corpus luteal cells obtained from female crossbred Taiwan native goats in the estrous cycle; the observation was performed using hematoxylin and eosin and immunohistochemistry staining. Subsequently, we used kisspeptin-10 (Kp-10) to stimulate temperature sensitive-caprine luteal cell line (ts-CLC-D) cells to investigate the progesterone (P4) levels, steroidogenic messenger RNA (mRNA)/protein levels, cell survival rate, intracellular Ca2+ concentration, and cell proliferation-related mRNA/protein levels in the mitogen-activated protein kinase pathway in vitro by applying immunofluorescence staining, Western blotting, 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide assay, and real-time polymerase chain reaction. RESULTS We observed the presence of proteins and mRNAs for STAR, CYP11A1, HSD3B, KISS1, and KISS1R in the corpus luteal cells from goats in vivo. In vitro, the addition of Kp-10 reduced the P4 levels (p < 0.01) and increased cell proliferation (p < 0.05) of the ts-CLC-D cells. Furthermore, we found that the levels of proteins and mRNA for STAR, CYP11A1, and HSD3B decreased significantly when Kp-10 was added (p < 0.05). However, adding Kp-10 did not affect the mRNA levels for PLCG2, DAG1, PRKCA, KRAS, RAF1, MAP2K1, MAP2K2, MAPK3, MAPK1, and MAPK14. CONCLUSION We determined that KISS1 could affect the P4 levels, steroidogenesis, and cell proliferation in luteal cells. However, further research is required to clarify how KISS1 regulates proliferation and steroid production in luteal cells.

Background: The percentage of female medical students has been significant elevating worldwide. The demographic shift is expected to influence the proportion of male versus female surgeons soon. The objective of this study was to evaluate the gender differences in the acquisition of robotic suturing skills. Methods: We compared the robotic suturing performance between 39 male and 19 female medical students. We separated the training into two parts: phase I, involving virtual reality (VR) robotic simulation, and phase II, involving robotic dry-laboratory simulation training. Participants first conducted step-by-step exercises on the VR robotic simulator and then the robotic skin-suturing pad using the da Vinci robot. Results: The metric analysis of the VR task “suture sponge” showed that female students required less time (difference: −170.7 seconds, 95% CI: −247.4 to −94.0) and had fewer errors (error difference: −50, 95% CI: −74.2 to −25.8) to complete the suture sponge exercise compared to male students. Moreover, female students completed more stitches than male students (differences in mean stitch achieved: .35; 95% CI: .06 to .65). However, there was no difference in the quality scores of stitches by gender (p = 0.85). Conclusion: Female medical students performed better in the VR task of suture spongy and achieved more stitches than male students with the da Vinci system despite no difference in robotic suture quality by gender. Because this is the first study comparing gender performance on a robotic platform, further studies are required to investigate if different training approaches will affect the performance by gender. Keywords: Robotic surgery, Simulation, Gender difference, Virtual reality, Medical education

Various bio-applications of mesoporous materials (e.g., the immobilization of enzymes or the delivery of biomolecules such as siRNA) require large pores for the successful adsorption of the rather large molecules of interest and protecting the fragile cargo from external forces such as degradation. We describe the facile synthesis of functionalized mesoporous silica nanoparticles with large pores (LP-MSNs) providing high loading capacity for the immobilization of two differently-sized enzymes. The synthesis procedure yields homogeneous core–shell particles of about 100 nm in size with large mesopores (about 7 nm in diameter) and an azide-functionality inside the pores. The LP-MSNs were synthesized employing a co-condensation approach with the rather large micellar template cetyltrimethylammonium p-toluenesulfonate (CTATos). Due to the azide functionality, the LP-MSNs are suitable for bio-orthogonal click chemistry reactions within the porous network. Two different acetylene-functionalized enzymes (sp-carbonic anhydrase (CA) and sp-horseradish peroxidase (HRP)) were immobilized in the pores of the obtained LP-MSNs by a copper-catalyzed 1,3-dipolar cycloaddition reaction. The covalent attachment of the enzymes within the mesopores allowed us to investigate the catalytic performance of the enzyme–silica systems. The enzymes are stable after bioconjugation with the silica support and show high catalytic activity over several cycles for the colorimetric reaction of guaiacol (2-methoxyphenol) in case of LP-MSN–HRP and the hydrolysis of 4-nitrophenyl acetate (NPA) by LP-MSN–CA.

BACKGROUND Kisspeptin and its receptor KISS1R have been found to be essential regulators of reproductive function. Previous data have revealed the presence of Kiss1 and Kiss1r mRNAs in the hypothalamus and the testis of humans and rodents. However, the precise location and possible physiological role of the kisspeptin/KISS1R system in the testis remain ambiguous. METHODS We first produced an anti-KISS1R immunoglobulin Y antibody for KISS1R identification. To detect the exact sites of KISS1R and kisspeptin expression in the testis, we conducted immunohistochemistry assays on sections of testes. We used real-time polymerase chain reactions to identify Kiss1r in mice and to determine the expression levels of testicular genes. Finally, to verify the upstream regulation on the Kisspeptin/KISS1 receptor system, we treated primary mouse Leydig cells and MA-10 cells with luteinizing hormone (LH) and Br-cAMP, respectively, and examined Kiss1 and Kiss1r mRNA expression. RESULTS Immunohistochemistry assays revealed that kisspeptin was expressed in Leydig cells and KISS1R was localized in the seminiferous tubules. With real-time polymerase chain reactions, we found Kiss1r mRNA was constitutively expressed in the mouse testis from birth until the postnatal fourth week. Furthermore, mRNA expression of Kiss1 was synchronized with that of Insl3 and Cyp19a. However, the expression of the LH receptor-encoding gene increased 1 week earlier than did Kiss1 expression. This indicated that the kisspeptin/KISS1R system in the testis may be controlled by LH and cAMP signaling pathways. Finally, we confirmed that Kiss1 mRNA expression was increased in both LH-treated primary Leydig cells and Br-cAMP-treated MA-10 cells (p

The fission and fusion processes determine mitochondrial morphology and function, in which signaling lipids play pivotal roles. Phosphatidic acid (PA) generated by PLD6, a mitochondrial phospholipase D, is essential for mitochondrial fusion; however, the underlying mechanism is still unclear. We previously discovered that a nucleoside diphosphate kinase, NME3, is critical for mitochondrial fusion yet through a kinase-independent activity. Here we demonstrate that NME3 functions as a mitochondrial tethering factor through binding to PA with its N-terminal amphipathic helix, enriching at the tip of mitochondria, and clustering membranes via its oligomeric structure. Optogenetic induction of PLD activity enhances NME3 recruitment to mitochondria in a catalytic-dependent manner, and NME3 is required for PLD6-mediated mitochondrial fusion. Together, PLD6-generated PA on mitochondria promotes NME3 binding, thereby increasing mitochondrial tethering to facilitate membrane fusion.

Background Issuing of correct prescriptions is a foundation of patient safety. Medication errors represent one of the most important problems in health care, with ‘look-alike and sound-alike’ (LASA) being the lead error. Existing solutions to prevent LASA still have their limitations. Deep learning techniques have revolutionized identification classifiers in many fields. In search of better image-based solutions for blister package identification problem, this study using a baseline deep learning drug identification (DLDI) aims to understand how identification confusion of look-alike images by human occurs through the cognitive counterpart of deep learning solutions and thereof to suggest further solutions to approach them. Methods We collected images of 250 types of blister-packaged drug from the Out-Patient Department (OPD) of a medical center for identification. The deep learning framework of You Only Look Once (YOLO) was adopted for implementation of the proposed deep learning. The commonly-used F1 score, defined by precision and recall for large numbers of identification tests, was used as the performance criterion. This study trained and compared the proposed models based on images of either the front-side or back-side of blister-packaged drugs. Results Our results showed that the total training time for the front-side model and back-side model was 5 h 34 min and 7 h 42 min, respectively. The F1 score of the back-side model (95.99%) was better than that of the front-side model (93.72%). Conclusions In conclusion, this study constructed a deep learning-based model for blister-packaged drug identification, with an accuracy greater than 90%. This model outperformed identification using conventional computer vision solutions, and could assist pharmacists in identifying drugs while preventing medication errors caused by look-alike blister packages. By integration into existing prescription systems in hospitals, the results of this study indicated that using this model, drugs dispensed could be verified in order to achieve automated prescription and dispensing.

Large-pore mesoporous silica nanoparticles (LP-MSNs) with defined particle size (

Paraquat, an herbicide used extensively worldwide, can cause severe toxicity in humans and animals, leading to irreversible, lethal lung fibrosis. The potential of CO-releasing molecules (CORMs), substances that release CO (Carbon monoxide) within animal tissues, for treating paraquat-induced ROS generation and inflammation is investigated here. Our results show that the fast CO releaser CORM-3 (4-20 μM) acts as a potential scavenger of free radicals and decreases fibrosis progression by inhibiting paraquat-induced overexpression of connective tissue growth factor and angiotensin II in MRC-5 cells. The slow CO releaser CORM-A1 (5 mg/kg) clearly decreased expression of the lung profibrogenic cytokines COX-2, TNF-α, and α-SMA and serum hydroxyproline, resulting in a lower mortality rate in paraquat-treated mice. Mice treated with higher-dose CORM-A1 (10 mg/kg) had relatively intact lung lobes and fewer fibrotic patches by gross observation, with less collagen deposition, mesangial matrix accumulation, and pulmonary fibrosis resulting from the mitigation of TGF-β overexpression. In conclusion, our data demonstrate for the first time that CORM-A1 alleviated the development of the fibrotic process and improved survival rate in mice exposed to PQ, would be an attractive therapeutic approach to attenuate the progression of pulmonary fibrosis following PQ exposure.

A dual surfactant synthetic approach was employed for the synthesis of large-pore mesoporous silica nanoparticles (MSNs). The as-synthesized mesoporous silica exhibited not only 2D hexagonal mesostructure with an average pore size of about 7 nm but also a spherical nanoparticulate morphology. A delayed co-condensation synthesis method was further employed to create site-specific core-shell bifunctional MSNs. These core-shell large-pore MSNs are biocompatible and can be efficiently taken up by cells. They are therefore promising nanocarriers for cellular delivery purposes.

Patients with lung cancer harboring activating epidermal growth factor (EGFR) mutations and pre-existing diabetes have been demonstrated to exhibit poor responses to first-line EGFR-tyrosine kinase inhibitor (TKI) therapy. Strategies for the management of acquired resistance to EGFR-TKIs in patients with advanced non-small cell lung cancer (NSCLC) are urgently required. Only a limited number of studies have been published to date on the effects of insulin on EGFR-TKI resistance in NSCLC. Hence, the aim of the present study was to investigate the roles of hyperinsulinemia and hyperglycemia in mediating gefitinib resistance in NSCLC cells with activating EGFR mutations. In the present study, the HCC4006 cell line, which harbors EGFR mutations, was co-treated with gefitinib and long-acting insulin glargine. Whether hyperinsulinemia is able to mediate EGFR-TKI resistance in the NSCLC cell line harboring activating EGFR mutations was also investigated, and the possible underlying mechanisms responsible for these actions were explored. The inhibition of cell proliferation, and the potential mechanism of gefitinib resistance, were examined using an MTS proliferation assay and western blot analysis, and through the transfection of siRNAs. Whether the inhibition of AKT is able to overcome EGFR-TKI resistance induced by long-acting insulin was also investigated. The results obtained suggested that hyperinsulinemia induced by glargine upregulated NSCLC cell proliferation and survival, and induced gefitinib resistance. By contrast, the morphology and proliferation of the cells in a medium containing a 2-fold concentration of glucose were not significantly affected. Gefitinib resistance induced by hyperinsulinemia may have been mediated via the phosphoinositide 3-kinase (PI3K)/AKT pathway rather than the mitogen-activated protein kinase extracellular signal regulated kinase (MAPK/ERK) pathway. AKT serine/threonine kinase 1 knockdown by siRNA rescued the gefitinib resistance that was induced by hyperinsulinemia. In conclusion, hyperinsulinemia, but not hyperglycemia, was identified to cause the development of gefitinib resistance in NSCLC cells with activating EGFR mutations. However, additional studies are required to investigate strategies, such as co targeting hyperinsulinemia and the PI3K/AKT pathway, for overcoming EGFR-TKI resistance in patients with NSCLC.

Objective Peer-assisted learning has been regarded as an adjunct to teaching modalities. It remains inconclusive regarding the benefits of peer observation in skills learning. Hence, we investigated whether the active engagement (AE) of peer observation in addition to expert demonstration would facilitate the performance in the virtual reality (VR) tasks. Setting/Design The programs involved 4 VR tasks including basic (camera targeting), intermediate (energy dissection and energy switching), and advanced (suture sponge) tasks in the da Vinci Skills Simulators, which were set up in the operating room at Taipei Medical University Hospital. Fifty medical students participated in the study. The AE of the participants was defined as the total number of peer observations in addition to expert observation before their performance. We assessed the correlations between AE and surgical task performance using Pearson correlation and the concept of learning analytics. Participants Medical students (sixth-year students in Taiwan, equivalent to fourth-year students in the US system) from Taipei Medical University were recruited. Results AE was correlated with the energy dissection task (r = 0.329, p = 0.02) and marginally associated with the energy switching task (r = 0.271, p = 0.057). However, AE was not correlated with either task scores for camera targeting (r = 0.096, p = 0.509) or task scores for suture sponge (r = −0.091, p = 0.529). Conclusions Our findings suggest that AE of peer observation may facilitate learning energy dissection task, which is an intermediate-level task, but not in other basic or advanced tasks in a VR context. The study highlights the potential effect of AE of peer observation on surgical learning based on a distinct level of tasks. Tasks that fit the learners’ level are recommended. Nevertheless, the effectiveness of peer observation on surgical training still has to be explored to ensure favorable results and optimal learning outcomes.

Enterovirus 71 (EV71) causes the outbreaks of hand-foot-and-mouth disease and results in deaths of hundreds of young children. EV71 virus-like particles (VLPs) are empty capsids consisting of viral structural proteins and can elicit potent immune responses, thus holding promise as an EV71 vaccine candidate. However, an efficient, scalable production and purification scheme is missing. For mass production of EV71 VLPs, this study aimed to develop a production and chromatography-based purification process. We first demonstrated the successful EV71 VLPs production in the stirred-tank bioreactor in which High Five™ cells were infected with a recombinant baculovirus co-expressing EV71 structural polyprotein P1 and protease 3CD. The culture supernatant containing the VLPs was subjected to tangential flow filtration (TFF) for concentration/diafiltration, which enabled the removal of >80% of proteins while recovering >80% of VLPs. The concentrated VLPs were next subjected to hydroxyapatite chromatography (HAC) in which the VLPs were mainly found in the flow through. After another TFF concentration/diafiltration, the VLPs were purified by size-exclusion chromatography (SEC) and concentrated/diafiltered by a final TFF. The integrated process yielded an overall VLPs recovery of ≈ 36% and a purity of ≈ 83%, which was better or comparable to the recovery and purity for the purification of live EV71 virus particles. This process thus may move the EV71 VLPs vaccine one step closer to the clinical applications.

Patients with lung cancer harboring activating epidermal growth factor (EGFR) mutations and pre-existing diabetes have been demonstrated to exhibit poor responses to first-line EGFR-tyrosine kinase inhibitor (TKI) therapy. Strategies for the management of acquired resistance to EGFR-TKIs in patients with advanced non-small cell lung cancer (NSCLC) are urgently required. Only a limited number of studies have been published to date on the effects of insulin on EGFR-TKI resistance in NSCLC. Hence, the aim of the present study was to investigate the roles of hyperinsulinemia and hyperglycemia in mediating gefitinib resistance in NSCLC cells with activating EGFR mutations. In the present study, the HCC4006 cell line, which harbors EGFR mutations, was co-treated with gefitinib and long-acting insulin glargine. Whether hyperinsulinemia is able to mediate EGFR-TKI resistance in the NSCLC cell line harboring activating EGFR mutations was also investigated, and the possible underlying mechanisms responsible for these actions were explored. The inhibition of cell proliferation, and the potential mechanism of gefitinib resistance, were examined using an MTS proliferation assay and western blot analysis, and through the transfection of siRNAs. Whether the inhibition of AKT is able to overcome EGFR-TKI resistance induced by long-acting insulin was also investigated. The results obtained suggested that hyperinsulinemia induced by glargine upregulated NSCLC cell proliferation and survival, and induced gefitinib resistance. By contrast, the morphology and proliferation of the cells in a medium containing a 2-fold concentration of glucose were not significantly affected. Gefitinib resistance induced by hyperinsulinemia may have been mediated via the phosphoinositide 3-kinase (PI3K)/AKT pathway rather than the mitogen-activated protein kinase extracellular signal regulated kinase (MAPK/ERK) pathway. AKT serine/threonine kinase 1 knockdown by siRNA rescued the gefitinib resistance that was induced by hyperinsulinemia. In conclusion, hyperinsulinemia, but not hyperglycemia, was identified to cause the development of gefitinib resistance in NSCLC cells with activating EGFR mutations. However, additional studies are required to investigate strategies, such as co targeting hyperinsulinemia and the PI3K/AKT pathway, for overcoming EGFR-TKI resistance in patients with NSCLC.

The aims of this work were to assess dental age using the Demirjian system and to evaluate the discrepancy between chronological and dental age among children residing in Western Australia. Forty panoramic radiographs of 5- to 11-year-old children taken at the paediatric clinic of a dental hospital in Western Australia were selected. The developmental stage of each left permanent mandibular tooth excluding the third molar was assessed by three trained examiners. Estimated dental age using the Demirjian system was compared with the chronological age of each child. The accuracy of the Demirjian system in this group of children was 42.5%. The difference between chronological age and dental age was significant (t = 4.066, df = 39, p < 0.001). Dental age estimation for Caucasian children had greater accuracy (χ2 = 5.013, df = 1, p = 0.025). Gender and orthodontic concerns were not related to the accuracy of the Demirjian system (p ≥ 0.481). The discrepancy between chronological and dental age using the Demirji...

To evaluate the effectiveness of a simulation-based flipped classroom in gaining the laparoscopic skills in medical students.An intervention trial.Taipei Medical University Hospital, an academic teaching hospital.Fifty-nine medical students participating in a 1-hour laparoscopic skill training session were randomly assigned to a conventional classroom (n = 29) or a flipped classroom approach (n = 30) based on their registered order. At the end of the session, instructors assessed participants' performance in laparoscopic suturing and intracorporeal knot-tying using the assessment checklist based on a modified Objective Structured Assessment of Technical Skills tool.Students in the flipped group completed more numbers of stitches (mean [M] = 0.47; standard deviation [SD] = 0.507) than those in the conventional group (M = 0.10; SD = 0.310) (mean difference: 0.37; 95% CI: 0.114-582; p = 0.002). Moreover, students in the flipped group also had higher stitch quality scores (M = 7.17; SD = 2.730) than those in the conventional group (M = 5.14; SD = 1.767) (mean difference = 2.03; 95% CI: 0.83-3.228; p = 0.001). Meanwhile, students in the flipped group had higher pass rates for the second throw (p0.001), third throw (p = 0.002), appropriate tissue reapproximation without loosening or strangulation (p0.001), needle cut from suture under direct visualization (p = 0.004), and needle safely removed under direct visualization (p = 0.018) than those in the conventional group.Comparing with traditional approach, a simulation-based flipped classroom approach may improve laparoscopic intracorporeal knot-tying skill acquisition in medical students.

10532 Background: Evidence-based medicine (EBM) requires applying literature evidence to inform practice. Students from Taipei Medical University Hospital, trained in EBM concepts, participated in a preliminary study using Watson for Oncology (WfO), an evidence-based decision-support system to enhance the EBM skills of medical students. Methods: A class of 50 medical students compared traditional search methods (TSM) and WfO in a workshop divided into 2 sequential sessions on colon and lung cancer, respectively. All students were trained on WfO, and 2 groups of 25 students each were randomly assigned to either TSM or WfO in the first session. Those groups were then assigned to the alternate approach in the second session. Students completed a profile that included their clinical experience with each cancer type. Students used either WfO or TSM to help answer a series of questions related to colon or lung cancer. Students then completed a survey of attitudes towards the technology, followed by a constructed-response learning assessment without the aid of TSM or WfO. Assessments were scored and results compared using a Mann-Whitney U Test; outcomes at two different experience levels, based on student profiles, were compared using a Kruskal-Wallis test. Results: In this preliminary study, more than 70% of students reported limited clinical experience with either cancer. On the colon cancer assessment, students in the WfO group performed significantly better than the TSM group ( p = 0.0001); there was no significant difference detected for lung cancer. Students with more clinical experience felt that TSM was easier to learn than WfO ( p= 0.005); students with less experience felt that WfO was clearer and more understandable than TSM ( p= 0.002). Conclusions: These preliminary results are consistent with better learning outcomes for students using WfO in the colon cancer module. Students with more clinic experience reported that TSM was easier to learn than WfO, however it is unknown if this might be due to a potentially greater familiarity with TSM in this more experienced group. More studies are needed to determine what features, if any, of WfO can facilitate EBM approaches in oncology education.

Adipose-derived stem cells (ASCs) have captured growing interests for cartilage regeneration. Although ASCs chondrogenesis can be stimulated by genetic modification, whether genetically engineered ASCs hold promise for the cartilaginous tissue formation remains to be explored. Since baculovirus (an emerging gene delivery vector) effectively transduced ASCs and transforming growth factor β3 (TGF-β3) was recently shown to induce ASCs chondrogenesis more potently than TGF-β1, we constructed a baculoviral vector (Bac-CT3W) to encode TGF-β3. The Bac-CT3W-transduced ASCs expressed TGF-β3 robustly and substantiated the chondrogenesis of ASCs cultured in monolayer and in porous scaffolds. Culture of the transduced cell/scaffold constructs in the rotating-shaft bioreactor (RSB) under hypoxic and perfusion conditions for 2 weeks further augmented the ASCs chondrogenesis and deposition of cartilage-specific collagen II and glycosaminoglycans, leading to the formation of cartilage-like tissues with hyaline appearance and compressive modulus approaching 62% of the native articular cartilage. Intriguingly, prolonged culture to 3 or 4 weeks failed to further augment the construct growth, probably due to the scaffold degradation. Altogether, baculovirus-mediated TGF-β3 expression in ASCs in conjunction with dynamic culture in the RSB for 2 weeks synergistically ameliorated the ASCs chondrogenesis and formation of cartilaginous tissues, representing a novel approach to producing engineered cartilages.

Chromobodies have recently drawn great attention as bioimaging nanotools. They offer high antigen binding specificity and affinity comparable to conventional antibodies, but much smaller size and higher stability. Chromobodies can be used in live cell imaging for specific spatio-temporal visualization of cellular processes. To date, functional application of chromobodies requires lengthy genetic manipulation of the target cell. Here, we develop multifunctional large-pore mesoporous silica nanoparticles (MSNs) as nanocarriers to directly transport chromobodies into living cells for antigen-visualization in real time. The multifunctional large-pore MSNs feature high loading capacity for chromobodies and are efficiently taken up by cells. By functionalizing the internal MSN surface with nitrilotriacetic acid-metal ion complexes, we can control the release of His6-tagged chromobodies from MSNs in acidified endosomes and observe successful chromobody-antigen binding in the cytosol. Hence, by combining the two nanotools, chromobodies and MSNs, we establish a new powerful approach for chromobody applications in living cells.

To develop the enterovirus 71 (EV71) vaccine, we previously constructed a recombinant baculovirus (Bac-P1-3CD) co-expressing EV71 P1 (under polyhedrin promoter) and 3CD (under p10 promoter) proteins, which caused P1 cleavage by 3CD protease and self-assembly of virus-like particles (VLPs) in Sf-9 cells. Assuming that reducing the 3CD expression can alleviate the competition with P1 expression and elevate the VLPs yield, hereby we constructed Bac-P1-C3CD and Bac-P1-I3CD expressing 3CD under weaker CMV and IE-1 promoters, respectively. Western blot and ELISA analyses revealed that Bac-P1-C3CD and Bac-P1-I3CD led to the VLPs release into the supernatant and enhanced the extracellular VLPs yield in Sf-9 cells, but gave poor VLPs production in High Five™ (Hi-5) cells. By optimizing the process parameters including host cells, cell density, culture mode and dissolved oxygen (DO), the best extracellular VLPs yield was achieved by infecting Sf-9 cells (4 × 10(6)cells/mL) cultured in the bioreactor (DO=30%) with Bac-P1-C3CD, which approached ≈64.3mg/L and represented a ≈43-fold increase over the yield (1.5mg/L) attained using the old process (Bac-P1-3CD infection of Sf-9 cells in the spinner flasks). The resultant VLPs not only resembled the VLPs produced from Bac-P1-3CD infection in density, size and shape, but also induced potent antibody responses in mouse models. The antibodies neutralized EV71 strains of homologous and heterologous genogroups, implicating the potential of the VLPs to confer cross-protection for the prevention of future epidemics. Altogether, Bac-P1-C3CD and the bioprocess render mass production more economical, obviate the need for cell lysis and hold promise for future industrial vaccine production.

High-performance self-aligned inversion-channel In 0.75 Ga 0.25 As n -MOSFETs using in situ ultra-high-vacuum (UHV) deposited Al 2 O 3 /Ga 2 O 3 (Gd 2 O 3 ) [GGO] and ex-situ atomic-layer-deposited (ALD) Al 2 O 3 as gate dielectrics have been fabricated. Both devices exhibit high drain currents and transconductances. A 1.2 μm-gate-length In 0.75 Ga 0.25 As MOSFET using Al 2 O 3 (2 nm-thick)/GGO (13 nm-thick) gate dielectric demonstrated a maximum drain current of 970 μA/μm, a peak transconductance of 410 μS/μm, and a peak mobility of 1560 cm 2 /V s. A maximum drain current of 740 μA/μm and a peak transconductance of 325 μS/μm were exhibited by a 1 μm-gate-length In 0.75 Ga 0.25 As MOSFET using ALD-Al 2 O 3 (6 nm-thick). A comparison between the inversion-channel InGaAs MOSFETs with gate dielectrics using the UHV- and ALD-approaches, and fabricated using the same self-aligned process, was carried out to provide insights for achieving InGaAs MOSFETs with even higher device performances. The comparison in the device performances was extended to cover representative enhancement-mode InGaAs MOSFETs, including self-aligned inversion-channel, non-self-aligned inversion-channel, and flat-band, or buried channel-type of E-mode (non inversion-channel) III–V devices.

Baculovirus is an effective vector for gene delivery into various mammalian cells, including chondrocytes and mesenchymal stem cells, and has been employed for diverse applications. By gene delivery and expression of the growth factor, recombinant baculovirus has been shown to modulate the differentiation state of the cells and stimulates the production of extracellular matrix and tissue formation, hence repairing the damaged cartilage and bone in vivo. This article reviews the studies pertaining to the applications of baculovirus-mediated gene delivery in cartilage and bone tissue engineering and discusses recent progress, future applications and potential hurdles.

This work proposes a new gateless one-time programmable (OTP) cell. This gateless OTP cell has a parasitic oxide–nitride–oxide (ONO) structure as the storage node and is successfully demonstrated in a 45 nm CMOS logic process. This gateless OTP cell, formed in a pure logic process and decoupled from gate oxide, is highly stable with a five orders of on/off current window. It also exhibits superior program performance, with an operating voltage of only 5 V and at a programming current of no more than 10 μA. Unlike breakdown-based anti-fuses, electron trapping of the gateless OTP cell demonstrates repeatability for testing. The gateless OTP can be UV-erased and is stable over 10 P/E cycles. An electrical erase mechanism with limited performance is also characterized and discussed. This new nitride gateless anti-fuse cell is a very promising logic OTP solution that provides fully compatibility to CMOS process below the 90 nm node.

Purpose – The purpose of this paper is to examine evaluations of teaching from viewpoints of lecturers and students to reveal perceived differences in teaching performance.Design/methodology/approach – A survey is administered to lecturers and students in a large‐scale university in Taiwan. Valid samples include ten lecturers and 250 students. Upon obtaining consensuses from lecturers, 25 students in each course are randomly selected to fill out questionnaires. Lecturers are requested to fill out questionnaires designed for instructors only. Factor, cluster, and multivariate analyses of variance are applied in analyzing the survey data. Five dimensions of evaluations of teaching are extracted from students' viewpoints, including content of materials, learning condition, interaction, attitudes, and responsiveness. These five factors are used in the clustering procedure to segment courses into superior‐ and inferior‐quality.Findings – Results indicate surveyed students in the clusters of superior‐ and infer...

Mucin glycoproteins are key components of native mucus which serves as an initial barrier in the human body against microbial attack. Mucins are able to prevent bacterial adhesion and can trap viruses. However, the weak mechanical properties of mucin solutions have so far prevented their application in a physiological environment. Here, methylcellulose biopolymers are used as mechanical adjuvants to overcome this limitation and generate a thermoresponsive mucin/methylcellulose hybrid system. The hybrid material developed combines the selective permeability properties brought about by mucins with the thermal autogelation properties of methylcellulose. As a consequence, triggered by contact with body-warm surfaces, the hybrid material rapidly forms a gel at physiological conditions, and this external temperature stimulus can also be harnessed to stimulate drug release from incorporated thermosensitive liposomes. Finally, the hybrid gel selectively retards the release of embedded molecules which can be used to further control and prolong drug release from the material.

ABSTRACT ABSTRACT ABSTRACT: The purpose of this research was to improve probiotic microencapsulation using prebiotics and : The purpose of this research was to improve probiotic microencapsulation using prebiotics and : The purpose of this research was to improve probiotic microencapsulation using prebiotics and : The purpose of this research was to improve probiotic microencapsulation using prebiotics and : The purpose of this research was to improve probiotic microencapsulation using prebiotics and modern optimization techniques to determine optimal processing conditions, performance, and survival rates. modern optimization techniques to determine optimal processing conditions, performance, and survival rates. modern optimization techniques to determine optimal processing conditions, performance, and survival rates. modern optimization techniques to determine optimal processing conditions, performance, and survival rates. modern optimization techniques to determine optimal processing conditions, performance, and survival rates. Prebiotics (fructooligosaccharides or isomaltooligosaccharides), growth promoter (peptide), and sodium algi- Prebiotics (fructooligosaccharides or isomaltooligosaccharides), growth promoter (peptide), and sodium algi- Prebiotics (fructooligosaccharides or isomaltooligosaccharides), growth promoter (peptide), and sodium algi- Prebiotics (fructooligosaccharides or isomaltooligosaccharides), growth promoter (peptide), and sodium algi- Prebiotics (fructooligosaccharides or isomaltooligosaccharides), growth promoter (peptide), and sodium algi- nate were incorporated as coating materials to microencapsulate 4 probiotics ( nate were incorporated as coating materials to microencapsulate 4 probiotics ( nate were incorporated as coating materials to microencapsulate 4 probiotics ( nate were incorporated as coating materials to microencapsulate 4 probiotics ( nate were incorporated as coating materials to microencapsulate 4 probiotics (Lactobacillus acidophilus Lactobacillus acidophilus Lactobacillus acidophilus Lactobacillus acidophilus Lactobacillus acidophilus, Lacto- Lacto- Lacto- Lacto- bacillus casei bacillus casei bacillus casei bacillus casei bacillus casei, Bifidobacterium bifidum Bifidobacterium bifidum Bifidobacterium bifidum Bifidobacterium bifidum Bifidobacterium bifidum, and Bifidobacterium longum Bifidobacterium longum Bifidobacterium longum Bifidobacterium longum Bifidobacterium longum). The proportion of the prebiotics, pep- ). The proportion of the prebiotics, pep- ). The proportion of the prebiotics, pep- ). The proportion of the prebiotics, pep- ). The proportion of the prebiotics, pep- tide, and sodium alginate was optimized using response surface methodology (RSM) to 1st construct a surface tide, and sodium alginate was optimized using response surface methodology (RSM) to 1st construct a surface tide, and sodium alginate was optimized using response surface methodology (RSM) to 1st construct a surface tide, and sodium alginate was optimized using response surface methodology (RSM) to 1st construct a surface tide, and sodium alginate was optimized using response surface methodology (RSM) to 1st construct a surface model, with sequential quadratic programming (SQP) subsequently adopted to optimize the model and evaluate model, with sequential quadratic programming (SQP) subsequently adopted to optimize the model and evaluate model, with sequential quadratic programming (SQP) subsequently adopted to optimize the model and evaluate model, with sequential quadratic programming (SQP) subsequently adopted to optimize the model and evaluate model, with sequential quadratic programming (SQP) subsequently adopted to optimize the model and evaluate the survival of microencapsulated probiotics under simulated gastric fluid test. Optimization results indicated the survival of microencapsulated probiotics under simulated gastric fluid test. Optimization results indicated the survival of microencapsulated probiotics under simulated gastric fluid test. Optimization results indicated the survival of microencapsulated probiotics under simulated gastric fluid test. Optimization results indicated the survival of microencapsulated probiotics under simulated gastric fluid test. Optimization results indicated that 1% sodium alginate mixed with 1% peptide and 3% fructooligosaccharides as coating materials would that 1% sodium alginate mixed with 1% peptide and 3% fructooligosaccharides as coating materials would that 1% sodium alginate mixed with 1% peptide and 3% fructooligosaccharides as coating materials would that 1% sodium alginate mixed with 1% peptide and 3% fructooligosaccharides as coating materials would that 1% sodium alginate mixed with 1% peptide and 3% fructooligosaccharides as coating materials would produce the highest survival in terms of probiotic count. The verification experiment yielded a result close to the produce the highest survival in terms of probiotic count. The verification experiment yielded a result close to the produce the highest survival in terms of probiotic count. The verification experiment yielded a result close to the produce the highest survival in terms of probiotic count. The verification experiment yielded a result close to the produce the highest survival in terms of probiotic count. The verification experiment yielded a result close to the predicted values, with no significant difference ( predicted values, with no significant difference ( predicted values, with no significant difference ( predicted values, with no significant difference ( predicted values, with no significant difference (P > 0.05). The storage results also demonstrated that addition > 0.05). The storage results also demonstrated that addition > 0.05). The storage results also demonstrated that addition > 0.05). The storage results also demonstrated that addition > 0.05). The storage results also demonstrated that addition of prebiotics in the walls of probiotic microcapsules provided improved protection for the active organisms. of prebiotics in the walls of probiotic microcapsules provided improved protection for the active organisms. of prebiotics in the walls of probiotic microcapsules provided improved protection for the active organisms. of prebiotics in the walls of probiotic microcapsules provided improved protection for the active organisms. of prebiotics in the walls of probiotic microcapsules provided improved protection for the active organisms. These probiotic counts remained at 10 These probiotic counts remained at 10 These probiotic counts remained at 10 These probiotic counts remained at 10 These probiotic counts remained at 10 6 to 10 to 10 to 10 to 10 to 10 7 colony-forming units (CFU)/g for microcapsules stored for 1 mo colony-forming units (CFU)/g for microcapsules stored for 1 mo colony-forming units (CFU)/g for microcapsules stored for 1 mo colony-forming units (CFU)/g for microcapsules stored for 1 mo colony-forming units (CFU)/g for microcapsules stored for 1 mo and then treated in simulated gastric fluid test and bile salt test. and then treated in simulated gastric fluid test and bile salt test. and then treated in simulated gastric fluid test and bile salt test. and then treated in simulated gastric fluid test and bile salt test. and then treated in simulated gastric fluid test and bile salt test. K K K K Keywords: microencapsulation, prebiotics, probiotics, response surface methodology microencapsulation, prebiotics, probiotics, response surface methodology microencapsulation, prebiotics, probiotics, response surface methodology microencapsulation, prebiotics, probiotics, response surface methodology microencapsulation, prebiotics, probiotics, response surface methodology

Pollution of water and soil by heavy-metal byproducts of mining and industrial operations is an environmental problem in many parts of Taiwan. Bermudagrass (Cynodon dactylon) and seashore paspalum (Paspalum vaginatum Swartz) are two of the most saline-tolerant warm-season turfgrasses. A whole-plant microculture (WPMC) system has been developed that permits intact-root growth observation through a gelrite-solidified culture medium. To screen efficiently for potential whole-plant-level salt tolerance in these turfgrasses, root morphology was studied using scanning electron microscopy. Nodal explants at the same developmental stage as the plants were sterilized, and each explant was placed in a Pyrex test tube containing 25 mL of 1/2 Murashige and Skoog (MS) media with 30 g L− 1 sucrose, 1 mg L− 1 IBA, 0.1 mg 2ip, 2.5 g L− 1gelrite, and either 0, 1, or 100 ppm of CdCl2 stress media for five months. The experimental results demonstrated significant differences among stress media in terms of root numb...

Enterovirus 71 (EV71) is responsible for the outbreaks of hand-foot-and-mouth disease that caused significant mortality in children, but no vaccine is available yet. EV71 virus-like particle (VLP) is the empty capsid consisting of viral structural proteins but can elicit potent immune responses, rendering VLP a promising EV71 vaccine candidate. To evaluate whether VLP remains stable after long-term storage, which is crucial for advancing the VLP vaccine to the clinical setting, we evaluated the effects of NaCl concentration, buffers and temperatures on the VLP stability. We first validated the use of dynamic light scattering (DLS) for measuring the hydrodynamic diameter (≈30-35 nm) of VLP, which was close to the VLP diameter (≈25-27 nm) as measured by transmission electron microscopy (TEM). Using these techniques, we found that EV71 VLP remained stable for 5 months in sodium phosphate (NaPi) buffers with various NaCl concentrations. EV71 VLP also remained morphologically stable in NaPi, citrate and TE(+) buffers for 5 months, yet the enzyme-linked immunosorbent assay (ELISA) revealed that the VLP stored in citrate and TE(+) buffers partially lost the immunogenicity after 5 months. In contrast, the VLP stored in the NaPi buffer at 4°C remained stable macroscopically and microscopically for 5 months, as judged from the DLS, TEM and ELISA. The VLP stored at 25°C and 37°C also retained stability for 1 month, which would obviate the need of a cold chain during the shipping. These data altogether proved the stability of EV71 VLP and suggested that the VLP is amenable to bioprocessing and storage.

Epstein-Barr virus (EBV) belongs to the gammaherpesvirus family. To produce infectious progeny, EBV reactivates from latency into the lytic cycle by expressing the determinative lytic transactivator, Zta. In the presence of histone deacetylase inhibitor (HDACi), p53 is a prerequisite for the initiation of the EBV lytic cycle by facilitating the expression of Zta. In this study, a serial mutational analysis of Zta promoter (Zp) indicated an important role for the ZID element in responding to HDACi induction and p53 binds to this ZID element together with Sp1, a universal transcription factor. Abolition of the DNA-binding ability of Sp1 reduces the inducibility of ZID by HDACi and also reduces the amount of p53 binding to ZID. Finally, it was shown that EBV in p53-positive-lymphoblastoid cell lines (LCLs) can enter into the lytic cycle spontaneously; however, knockdown of p53 in LCLs leads to retardation of EBV reactivation.

Transduction of partially dedifferentiated rabbit chondrocytes with a baculovirus (Bac-CB) expressing bone morphogenetic protein-2 (BMP-2) reverses dedifferentiation and enhances matrix production. Hereby we examined whether transduction with Bac-CB in combination with another baculovirus expressing transforming growth factor-beta1 (TGF-beta1) or insulin-like growth factor-1 (IGF-1) synergistically augmented chondrogenic differentiation. Passage 3 rabbit articular chondrocytes were transduced by different baculovirus combinations: single transduction with Bac-CB, cotransduction with Bac-CB and Bac-CT (expressing TGF-beta1), cotransduction with Bac-CB and Bac-CI (expressing IGF-1), and transduction with Bac-CB followed by repeated transduction with Bac-CT, Bac-CI, or Bac-CB 5 days later. Transduced cells were encapsulated into alginate beads for culture. Among these strategies, only cotransduction with Bac-CB and Bac-CT led to improved redifferentiation when compared with Bac-CB single transduction, as evidenced by the enhanced expression of aggrecan and collagen IIB (Col IIB), suppressed expression of Col I and Col X, emergence of chondrocyte-specific lacunae, and elevated deposition of matrix molecules. The cotransduction also accelerated the expression of Sox9, Col IIB, and aggrecan. In summary, baculovirus-mediated coexpression of TGF-beta1 and BMP-2 synergistically accelerates the chondrocyte redifferentiation process and improves the maintenance of chondrocyte phenotype and accumulation of cartilage-specific matrix molecules.