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Overexpression of PD-L1 can be a predictive marker for anti-PD-1 therapeutic efficacy in classic Hodgkin lymphoma (CHL); however, harmonization of different IHC assays remains to be accomplished, and interpretations of PD-L1 immunostaining results remain controversial in CHL. In this study, we sought to optimize the PD-L1 immunohistochemistry (IHC) assay in CHL. All tests were performed on a tumour tissue microarray established from 54 CHL cases. Three IHC antibodies (405.9A11, SP142, 22C3) for detecting PD-L1 expression were compared semi quantitatively with the RNAscope assay (No. 310035, ACD), and the difference in the expression in background immune cells (ICs) between assays and the associations of expression levels with densities of TILs/TAMs were also analysed. 405.9A11 demonstrated best specificity in HRS cells and best sensitivity in ICs. Positive expression of PD-L1 was more frequent in ICs (85.2%) than in HRS cells (48.1%). Different subgroups of background ICs, including tumour-associated macrophages (TAMs), were assessed and scored for CD4, CD8, FOXP3, and CD163 expression. PD-L1 expression on ICs was the factor most associated with the density of TAMs. 405.9A11 provided the most convincing PD-L1 expression results. Pathologists should report PD-L1 expression in a combined manner, including both the status of HRS cells and the percentage of PD-L1-positive ICs. [ABSTRACT FROM AUTHOR]

Grandparental caregiving is common in China, which can directly impact grandchild development. In addition, grandparents may have an indirect impact on children through their relationship with and support of the parents. However, associations between grandparenting, parenting, and child outcomes are rarely investigated. The current study is a pilot study which includes 42 mothers, fathers, and co-residing grandparents in China, and examines whether grandparental sensitivity relates to parental sensitivity (biological child of the grandparent), and whether grandparental sensitivity and maternal and paternal sensitivity each uniquely contribute to 14-month-old children's Executive Function (EF). Results did not show a significant relation between parental and grandparental sensitivity or a unique contribution of sensitivity from mothers, fathers, or grandparents to infant EF. Our findings from exploratory analyses showed differences in sensitivity of grandparents and parents towards the third generation but did not confirm a combined effect of multiple caregivers on infant EF development in China. Highlights: This pilot study is the first to observe caregiving in mothers, fathers and co-residential grandparents in China. The study investigated the relation between grandparenting and parenting, and the association of multiple caregiving with infant EF development. Sensitivity levels of grandparents towards the third generation were lower than those of parents. There was no significant combined effect of multiple caregivers (mother, father and grandparent sensitivity) on infant EF development. [ABSTRACT FROM AUTHOR]

The present study investigated the efficacy and toxicity profile of first-line asparaginase (ASP)-based versus non-ASP-based regimens in treating early-stage extranodal NK/T-cell lymphoma (ENKTCL) in non-anthracycline therapy era. This multi-center, real-world retrospective study consisted 305 newly diagnosed localized ENKTCL patients who were treated with sequential chemoradiation between 2010 and 2020 in China: 190 cases received ASP-based regimens and 115 cases received non-ASP-based regimens. Propensity score matching and multivariable analyses were used to compare survivals and toxicities between the two treatment groups. Non-ASP-based regimens achieved comparable survivals compared with ASP-based regimens in the entire cohort. The 5-year overall survival (OS), progression-free survival (PFS) rates were 84.7% and 73.5% for non-ASP-based regimens, and 87.7% (P=0.464) and 74.6% (P=0.702) for ASP-based regimens. The non-inferior survivals of non-ASP-based regimens were consistent after adjustment using PSM and multivariable analyses. However, survival benefits of ASP varied in different treatment modalities. Among patients receiving sequential chemotherapy and radiation (CT+RT±CT), ASP-based regimens achieved higher complete remission rate (54.3 vs. 34.5%, P=0.047) and more favorable survivals compared with non-ASP-based regimens (5-year OS, 87.0 vs. 69.0%, P=0.028). However, for patients receiving sequential radiation and chemotherapy (RT+CT), non-ASP-based regimens achieved comparable favorable survivals as ASP-based regimens. Besides, liver injury, malnutrition, and coagulative dysfunction were significantly more commonly documented in ASP-based regimens. These findings suggested that ASP was an effective agent in treating ENKTCL, especially among those receiving induction CT and RT. For patients who received upfront RT, non-ASP-based regimens might be a comparably effective and more tolerable treatment option. [ABSTRACT FROM AUTHOR]

Visible particle identification is a crucial prerequisite step for process improvement and control during the manufacturing of injectable biotherapeutic drug products. Raman spectroscopy is a technology with several advantages for particle identification including high chemical sensitivity, minimal sample manipulation, and applicability to aqueous solutions. However, considerable effort and experience are required to extract and interpret Raman spectral data. In this study, we applied machine learning algorithms to analyze Raman spectral data for visible particle identification in order to minimize expert support and improve data analysis accuracy. We manually prepared ten types of particle standard solutions to simulate the particle types typically observed during manufacturing and established a Raman spectral library with accurate peak assignments for the visible particles. Five classification algorithms were trained using visible particle Raman spectral data. All models had high prediction accuracy of >98% for all types of visible particles. Our results demonstrate that the combination of Raman spectroscopy and machine learning can provide a simple and accurate data analysis approach for visible particle identification. [ABSTRACT FROM AUTHOR]

Intracellular pH plays a critical role in biological functions, and abnormal pH values are related to various diseases. Here, we report on an intracellular pH sensor AgInS2 (AIS)/ZnS quantum dots (QDs) that show long fluorescence lifetimes of hundreds of nanoseconds and low toxicity. Fluorescence lifetime imaging microscopy (FLIM) combined with AIS/ZnS QDs is used for the imaging of live cells in different pH buffers and different cell lines. The FLIM images of AIS/ZnS QDs in live cells demonstrate different intracellular pH values in different regions, such as in lysosomes or cytoplasm. This method can also distinguish cancer cells from normal cells, and the fluorescence lifetime difference of the AIS/ZnS QDs between the two types of cells is 100 ± 7 ns. Most importantly, the exfoliated cervical cells from 20 patients are investigated using FLIM combined with AIS/ZnS QDs. The lifetime difference value between the normal and cervical cancer (CC) groups is 115 ± 9 ns, and the difference between the normal and the precancerous lesion group is 64 ± 9 ns. For the first time, the noninvasive method has been used for cervical cancer screening, and it has shown great improvement in sensitivity compared with a clinical conventional cytology examination. [ABSTRACT FROM AUTHOR]

Simplicity and stability of an analytical method are the key to practical applications. In order to achieve such a goal, a molecule with multifunction could be one of the strategies. In this work, the multifunctional characteristics of branched polyethylenemine (bPEI) were employed for verification of this hypothesis. Three types of amines in the bPEI can be used to synthesize core-shell nanostructures (bPEI@Au and SiO2@bPEI@Au) in a simple one-pot process within 1 h, and as coreactants with over 500-fold enhancement for electrochemiluminescent (ECL) detection. Based on the remarkable performance of bPEI, a label-free ECL immunosensor and its array with the same assembly strategy were constructed for the simple and rapid determination of three acute myocardial infarction (AMI) biomarkers: cardiac troponin I (cTnI), human-type fatty acid binding protein (hFABP) and copeptin. Both methods exhibited excellent stability and sensitivity with wide linear range and low limits of detection down to fg/mL level by the photomultiplier tube and pg/mL level by the electron-multiplying charge-coupled device. [ABSTRACT FROM AUTHOR]

In this study, C-doped TiO2 nanoparticles (C-TiO2) were prepared and tested as a photosensitizer for visible-light-driven photodynamic therapy against cervical cancer cells (HeLa). X-ray diffraction and Transmission Electron Microscopy confirmed the anatase form of nanoparticles, spherical shape, and size distribution from 5 to 15 nm. Ultraviolet–visible light spectroscopy showed that C doping of TiO2 enhances the optical absorption in the visible light range caused by a bandgap narrowing. The photo-cytotoxic activity of C-TiO2 was investigated in vitro against HeLa cells. The lack of dark cytotoxicity indicates good biocompatibility of C-TiO2. In contrast, a combination with blue light significantly reduced the survival of HeLa cells: illumination only decreased cell viability by 30% (15 min of illumination, 120 µW power), and 60% when HeLa cells were preincubated with C-TiO2. We have also confirmed blue light-induced C-TiO2-catalyzed generation of reactive oxygen species in vitro and intracellularly. Oxidative stress triggered by C-TiO2/blue light was the leading cause of HeLa cell death. Fluorescent labeling of treated HeLa cells showed distinct morphological changes after the C-TiO2/blue light treatment. Unlike blue light illumination, which caused the appearance of large necrotic cells with deformed nuclei, cytoplasm swelling, and membrane blebbing, a combination of C-TiO2/blue light leads to controlled cell death, thus providing a better outcome of local anticancer therapy. [ABSTRACT FROM AUTHOR]

Reconfigurable micro–nanostructures have been largely used in the development of modern photocontrol technology. In this work, a dynamically adjustable hydrogel of supramolecule structures is realized by molecule self-assembly and cross-link in microscale. Calculations based on the molecule modeling systematically illuminate the assembly process and the mechanism of the dynamically adjustable micro–nanostructure. The transmittance changes of the hydrogel, which are measured in the visible-light range before and after electric heating, show outstanding light-control function. Thus, the one-step fabrication, bio-compatibility (no initiator, cross linker, or monomer residual), and agile photocontrol application demonstrate the potential of the adjustable physically bonded hydrogel in development of low-loss and integrated dynamical light-control devices. [ABSTRACT FROM AUTHOR]

TiO2 prolatenanospheroids (PNSs) may be photosensitizers (PSs), which act by catalyzation of hydroxyl radical (∙OH) formation upon light illumination. ∙OH might, in turn, contribute to killing of cancer cells. On the other hand, there is great concern about toxicity in the dark of TiO2 nanoparticles in general. In this work, we have investigated the biocompatibility of TiO2 PNSs of the anatase crystal form (length between 100 and 300 nm and width 50 nm) in the dark with immune cells and light-induced cytotoxicity on several cancer cell lines. The effects of the treatment of different cell lines with several concentrations of TiO2 PNSs suspensions showed the specifics of cells' viability and the intracellular localization. The results of in vitro studies obtained by cytotoxicity assays adjusted to individual cell lines' metabolism point towards the biocompatibility of TiO2 PNSs at low and moderate concentrations in the dark, which neither kill the cells, nor induce activation of the immune system cells. Laser scanning confocal microscopy revealed that PNSs are taken up by cells, and insight into the intracellular distribution was obtained in this study. [ABSTRACT FROM AUTHOR]

Negative birth experiences can lead to symptoms of post-traumatic stress disorder in new mothers but have received much less attention in new fathers. A sample of 314 first-time expectant couples rated their symptoms of anxiety and depression in the third trimester and at 4-month post birth (227 vaginal delivery, 87 caesarean section), when they also completed the emotional memories subscale of the BirthMARQ (Foley et al. BMC Pregnancy Childbirth, 14, 211, 2014). We first examined mode of delivery (vaginal birth versus caesarean section) as a predictor of mothers' and fathers' BirthMARQ scores. Next, we used actor-partner interdependence model (APIM) to investigate intra- and interpersonal associations between birth experiences and maternal/paternal latent factors for antenatal and postnatal depression/anxiety. Reports of negative birth experiences were more common for mothers than fathers and for parents of babies born by caesarean section than by vaginal delivery. Within-couple agreement was moderately strong and, for both parents at both time-points, individual differences in negative birth memories were associated with symptoms of depression and anxiety. Negative birth memories also played a mediating role in the association between birth via caesarean section and reduced postnatal maternal wellbeing. Given the striking similarities between mothers and fathers in links between birth experiences and wellbeing, our findings highlight the need for partner-inclusive intervention strategies. [ABSTRACT FROM AUTHOR]

Introduction: Mesenchymal stem cells (MSCs) are promising candidates for cell therapy owing to their therapeutic effect in various diseases. In general, MSCs grow efficiently in serum-containing culture media, indicating an essential role of adhesion in their mesenchymal lineage-specific propagation. Nevertheless, the use of non-human supplements in culture (xeno-free issue) in addition to the lack of control over unknown factors in the serum hampers the clinical transition of MSCs. Methods: In this study, embryonic stem cell derived mesenchymal stem cells (ES-MSCs) were used owing to their scalable production, and they expressed a series of MSC markers same as adipose-derived MSCs. The affinity of the culture matrix was increased by combining fibronectin coating with its adjuvant peptide, gelatin, or both (FNGP) on tissue culture polystyrene to compare the regenerative, therapeutic activities of ES-MSCs with a cell binding plate as a commercial control. Results: The FNGP culture plate promoted pivotal therapeutic functions of ES-MSCs as evidenced by their increased stemness as well as anti-inflammatory and proangiogenic effects in vitro. Indeed, after culturing on the FNGP plates, ES-MSCs efficiently rescued the necrotic damages in mouse ischemic hindlimb model. Conclusions: This study suggests a potential solution by promoting the surface affinity of culture plates using a mixture of human fibronectin and its adjuvant PHSRN peptide in gelatin. The FNGP plate is expected to serve as an effective alternative for serum-free MSC expansion for bench to clinical transition. [ABSTRACT FROM AUTHOR]

Angiogenesis induction into damaged sites has long been an unresolved issue. Local treatment with pro-angiogenic molecules has been the most common approach. However, this approach has critical side effects including inflammatory coupling, tumorous vascular activation, and off-target circulation. Here, the concept that a structure can guide desirable biological function is applied to physically engineer three-dimensional channel networks in implant sites, without any therapeutic treatment. Microchannel networks are generated in a gelatin hydrogel to overcome the diffusion limit of nutrients and oxygen three-dimensionally. Hydrogel implantation in mouse and porcine models of hindlimb ischemia rescues severely damaged tissues by the ingrowth of neighboring host vessels with microchannel perfusion. This effect is guided by microchannel size-specific regenerative macrophage polarization with the consequent functional recovery of endothelial cells. Multiple-site implantation reveals hypoxia and neighboring vessels as major causative factors of the beneficial function. This technique may contribute to the development of therapeutics for hypoxia/inflammatory-related diseases. Restoration of blood flow to damaged sites has commonly involved treatment with pro-angiogenic molecules but these have undesired side effects. Here the authors present a microchannel-patterned gelatin hydrogel that is able to rescue mouse and porcine models of hindlimb ischemia. [ABSTRACT FROM AUTHOR]

Purpose: To record the coexistence of polypoidal choroidal vasculopathy (PCV) with other rare fundus diseases in a Chinese population. Method: In this retrospective hospital-based study, a chart review of 861 patients with newly diagnosed PCV was performed. The clinical features of rare fundus comorbidities of PCV were recorded. Results: Five eyes of 5 patients aged 63.4 ± 11.22 years (0.58%) had PCV coexisting with other fundus diseases in the same eye. Of the 5 PCV patients, 2 (0.23%) had myelinated nerve fiber, 2 (0.23%) had branch retinal vein occlusion, and 1 (0.12%) had retinal angiomatous proliferation. Conclusion: We reported rare fundus comorbidities of PCV in a large Chinese cohort. These comorbidities included myelinated nerve fiber, branch retinal vein occlusion and retinal angiomatous proliferation. The combination might constitute an accidental occurrence. [ABSTRACT FROM AUTHOR]

The oblique-incidence reflectivity difference (OI-RD) scanning microscopy has the capability of simultaneously measuring binding curves of a protein probe with tens of thousands molecular targets in a microarray and yielding reaction rate constants. However, the quality of reaction rate constants is influenced by the fluidic system. To improve the quality of reaction rate constant measurement over the entire microarray, we demonstrate a fluidic chamber that allows the fluid to flow from the bottom to the top uniformly across the microarray and thus provides more uniform and accurate measurements of reaction rate constants with simplified fluidic design. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the changes of the photoreceptor layer (PRL) thickness with spectral domain optical coherence tomography (SD-OCT) and the retinal function by mfERG, as well as the correlation of morphology and function parameters in subjects with early and intermediate age-related macular degeneration (AMD). Methods: Subjects with clinical diagnosis of early or intermediate AMD and age-matched healthy subjects were recruited prospectively in this study. Color fundus photography, SD-OCT, and mfERG were conducted. Retinal photoreceptor thickness was measured, and first-order kernel responses were recorded. The differences between AMD group and control group were compared, and the correlations between macular photoreceptor thickness and the mfERG were analyzed. Results: PRL thickness (μm) in four areas including foveola and 0.5, 1.5, and 3 mm away from foveola was 192.48 ± 17.37, 163.73 ± 12.95, 130.93 ± 9.20, and 108.78 ± 7.81, respectively, in normal eyes, whereas in AMD group, they were 158.61 ± 45.25, 138.91 ± 20.92, 118.91 ± 12.85, and 95.00 ± 9.64, respectively ( P < 0.001). The mean amplitude response densities of AMD patients decreased significantly compared to the control group in ring 1-6 ( P < 0.001). The mean mfERG N1 and P1 latency of AMD patients prolonged compared to the control group, except the ring 1 ( P = 0.588 and P = 0.084). The macular PRL thickness was significantly associated with the mfERGN1 and P1 amplitude density in ring 1-4 ( r = 0.338-0.533, P < 0.01). Conclusions: PRL thickness decreases are in accordance with the deterioration of retinal electrophysiological activity. The retinal PRL thickness is important parameter to assess of early and intermediate AMD severity. [ABSTRACT FROM AUTHOR]

Transition metal manganese ion (Mn) doped zinc selenide quantum dots (Mn:ZnSe D-Dots) have been considered as a new material for fluorescent probes in biological labeling. However, this application is limited by the low membrane permeability of D-Dots. In this work, Mn:ZnSe D-Dots were capped with the polycation Sofast to label living cells. For the first time, the efficiency of cellular uptake in living cells is significantly enhanced. Various molar ratios of Sofast to D-Dots were explored and compared to obtain the optimal reaction conditions between Sofast and D-Dots for preparing Sofast/D-Dots nano-compound. A comparison on the fluorescence labeling ability of living cells were made between Sofast/D-Dots and pure D-Dots. Results from laser scanning confocal microscope show that Sofast/D-Dots complexes enter the cells more efficiently than pure D-Dots, even with a lower concentration and shorter incubation time. The cytotoxicities of D-Dots and Sofast/D-Dots were also studied. It was found that Sofast/D-Dots have a much lower cytotoxicity than cadmium-containing quantum dots (i.e. CdTe and CdTe/ZnS). Our results suggest that the non-heavy-metal-containing Sofast/D-Dots complexes have a great potential in the application of biological labeling, especially of long-time bioimaging in living cells. [ABSTRACT FROM AUTHOR]

Bovine serum albumin (BSA)-coated CdTe/ZnS quantum dots (BSA-QDs) were selected to conjugate with folic acid (FA), forming FA-BSA-QDs. This study aims to develop these small FA-BSA-QDs (less than 10 nm) for the diagnosis of cancers in which the FA receptor (FR) is overexpressed. The enhancement of cellular uptake in FR-positive human nasopharyngeal carcinoma cells (KB cells) for FA-BSA-QDs was found by means of confocal fluorescence microscopy under single-photon and two-photon excitation. The uptake enhancement for FA-BSA-QDs was further evaluated by flow-cytometric analysis in 10 KB cells, and was about 3 times higher than for BSA-QDs on average. The uptake enhancement was suppressed when KB cells had been pretreated with excess FA, reflecting that the enhancement was mediated by the association of FR at cell membranes with FA-BSA-QDs. When human embryonic kidney cells (293T) (FR-negative cells) and KB cells, respectively, were incubated with FA-BSA-QDs (1 μM) for 40 min, the FA-BSA-QD uptake by 293T cells was much weaker than that by KB cells, demonstrating that FA-BSA-QDs could undergo preferential binding on FR-positive cancer cells. These characteristics suggest that FA-BSA-QDs are potential candidates for cancer diagnosis. [ABSTRACT FROM AUTHOR]

The nuclear pore complex (NPC) is one of the largest supramolecular structures in eukaryotic cells. Its octagonal ring-scaffold perforates the nuclear envelope and features a unique molecular machinery that regulates nucleocytoplasmic transport. NPCs are composed of ~30 different nucleoporins (Nups), averaged at 8, 16 or 32 copies per NPC. This estimate has not been confirmed for individual NPCs in living cells due to the inherent difficulty of counting proteins inside single supramolecular complexes. Here we used single-molecule SPEED microscopy to directly count the copy-number of twenty-four different Nups within individual NPCs of live yeast, and found agreement as well as significant deviation from previous estimates. As expected, we counted 8 copies of four peripheral Nups and 16 copies of fourteen scaffold Nups. Unexpectedly, we counted a maximum of 16 copies of Nsp1 and Nic96, rather than 32 as previously estimated; and found only 10-15 copies of six other Nups, rather than 8 or 16 copies as expected. This in situ molecular-counting technology can test structure-function models of NPCs and other supramolecular structures in cells. [ABSTRACT FROM AUTHOR]