PD-L1-mediated immune evasion in triple-negative breast cancer is linked to the loss of ZNF652.

The intrinsic regulation of programmed death ligand-1 (PD-L1) expression remains unclear. Here, we report that zinc-finger protein 652 (ZNF652) is a potent transcription repressor of PD-L1. ZNF652 frequently experiences loss of heterozygosity (LOH) in various cancers. Higher LOH rate and lack of estrogen-inducible transcription lead to suppressed expression of ZNF652 in triple-negative breast cancer (TNBC). Mechanistically, ZNF652 is physically associated with the NuRD transcription co-repressor complex to repress a cohort of genes, including PD-L1. Overexpression of ZNF652 inhibits PD-L1 transcription, whereas depletion of ZNF652 upregulates PD-L1. Loss of ZNF652 in TNBC unleashes PD-L1-mediated immune evasion both in vitro and in vivo. Significantly, ZNF652 expression is progressively lost during breast cancer progression, and a low ZNF652 level is correlated with elevated PD-L1 expression, less infiltrated CD8+ T cells, and poor prognosis in TNBC. Our study provides insights into PD-L1 regulation and supports the pursuit of ZNF652 as a potential biomarker and drug target for breast cancer immunotherapy.

Modified mouse model of repeated mild traumatic brain injury through a thinned-skull window and fluid percussion.

Mild traumatic brain injury (mTBI) is a clinically highly heterogeneous neurological disorder, none of the existing animal models can replicate the entire sequelae. This study aimed to develop a modified closed head injury (CHI) model of repeated mTBI (rmTBI) for investigating Ca2+ fluctuations of the affected neural network, the alternations of electrophysiology, and behavioral dysfunctions. The transcranial Ca2+ study protocol includes AAV-GCaMP6s infection in the right motor cortex, thinned-skull preparation, and two-photon laser scanning microscopy (TPLSM) imaging. The CHI rmTBI model is fabricated using the thinned-skull site and applying 2.0 atm fluid percussion with 48-h interval. The neurological dysfunction, minor motor performance, evident mood, spatial working, and reference deficits we found in this study mimic the clinically relevant syndromes after mTBI. Besides, our study revealed that there was a trend of transition from Ca2+ singlepeak to multipeak and plateau, and the total Ca2+ activities of multipeaks and plateaus (p < .001 vs. pre-rmTBI value) were significantly increased in ipsilateral layer 2/3 motor neurons after rm TBI. In parallel, there is a low-frequency power shift from delta to theta band (p < .01 vs. control) in the ipsilateral layer 2/3 of motor cortex of the rmTBI mice, and the overall firing rates significantly increased (p < .01 vs. control). Moreover, rmTBI causes slight cortical and hippocampal neuron damage and possibly induces neurogenesis in the dentate gyrus (DG). The alternations of Ca2+ and electrophysiological characteristics in layer 2/3 neuronal network, histopathological changes, and possible neurogenesis may play concertedly and partially contribute to the functional outcome post-rmTBI.

Polarization-independent 3D metasurface with complex amplitude modulation.

Metasurfaces, which possess unprecedented capabilities in manipulating electromagnetic wavefronts, are promising for accurate complex amplitude modulation with a compact device. However, current strategy of complex amplitude modulation based on metasurfaces focuses on anisotropic unit design which is intrinsically constrained of polarization states. In this study, we propose a design methodology of polarization-independent metasurface which comprises an array of nanocylinders with various radii and heights. The effectiveness of the proposed scheme is verified using an optical vortex generator and a complex-amplitude hologram device. The straightforward, cost-effective, and polarization-independent design can provide robust and reliable solutions for wavefronts modulation in various optical applications.

3D printing of nanowrinkled architectures via laser direct assembly.

Structural wrinkles in nature have been widely imitated to enhance the surface functionalities of objects, especially three-dimensional (3D) architectured wrinkles, holding promise for emerging applications in mechanical, electrical, and biological processes. However, the fabrication of user-defined 3D nanowrinkled architectures is a long-pending challenge. Here, we propose a bottom-up laser direct assembly strategy to fabricate multidimensional nanowrinkled architectures in a single-material one-step process. Through the introduction of laser-induced thermal transition into a 3D nanoprinting process for leading the point-by-point nanoscale wrinkling and the self-organization of wrinkle structures, we have demonstrated the program-controlled and on-demand fabrication of multidimensional nanowrinkled structures. Moreover, the precise control of wrinkle morphology with an optimal wavelength of 40 nanometers and the regulation of the dynamic transformation of wrinkled cellular microstructures via interfacial stress mismatch engineering have been achieved. This study provides a universal protocol for constructing nearly arbitrary nanowrinkled architectures and facilitates a new paradigm in nanostructure manufacturing.

Rapid In-Situ Synthesis and Patterning of Edge-Unsaturated MoS2 by Femtosecond Laser-Induced Photo-Chemical Reaction.

Molybdenum disulfide (MoS2) is a representative transition metal sulfide that is widely used in gas and biological detection, energy storage, and integrated electronic devices due to its unique optoelectrical and chemical characteristics. To advance toward the miniaturization and on-chip integration of functional devices, it is strategically important to develop a high-precision and cost-effective method for the synthesis and integration of MoS2 patterns and functional devices. Traditional methods require multiple steps and time-consuming processes such as material synthesis, transfer, and photolithography to fabricate MoS2 patterns at the desired region on the substrate, significantly increasing the difficulty of manufacturing micro/nanodevices. In this work, we propose a single-step femtosecond laser-induced photochemical method which can realize the fabrication of arbitrary two-dimensional edge-unsaturated MoS2 patterns with high efficiency in microscale. Based on this method, MoS2 can be synthesized at a rate of 150 µm/s, 2 orders of magnitude faster than existing laser-based thermal decomposition methods without sacrificing the resolution and quality. The morphology and roughness of the MoS2 pattern can be controlled by adjusting the laser parameters. Furthermore, the femtosecond laser direct writing (FLDW) method was used to fabricate microscale MoS2-based gas detectors that can detect a variety of toxic gases with high sensitivity up to 0.5 ppm at room temperature. This FLDW method is not only applicable to the fabrication of high-precision MoS2 patterns and integrated functional devices, it also provides an effective route for the development of other micro/nanodevices based on a broad range of transition metal sulfides and other functional materials.

Cryopreservation of peripheral blood mononuclear cells using uncontrolled rate freezing.

Peripheral blood mononuclear cells are widely used as source material for anticancer immunotherapies. The conventional cryopreservation method for peripheral blood mononuclear cells is time-consuming and expansive, which involves controlled rate freezing followed by storage in liquid nitrogen. Instead, the convenient uncontrolled rate freezing cryopreservation method had been reported successfully in peripheral blood hematopoietic stem cells and peripheral blood progenitor cells. Therefore, we hypothesized that uncontrolled rate freezing cooling method maybe also applied to peripheral blood mononuclear cells cryopreservation. In this study, we evaluated the performance of uncontrolled rate freezing and controlled rate freezing cooling methods through cell recovery rate, viability, differentiation potential into cytokine-induced killer cells and the cellular properties of the cultured cytokine-induced killer cells. The results showed similar post-thaw viability and recovery rate in both controlled rate freezing and uncontrolled rate freezing cryopreserved peripheral blood mononuclear cells. Importantly, the uncontrolled rate freezing cryopreserved peripheral blood mononuclear cells exhibited higher growth ratio and earlier cell clustering during ex-vivo cytokine-induced killer cell culture than the controlled rate freezing ones. These two groups of expanded cytokine-induced killer cells also exhibited similar effector cell subset ratio and tumoricidal activity. In general, the performance of cryopreserved peripheral blood mononuclear cells using uncontrolled rate freezing cooling method, with the commercial cryoprotective agent CellBanker 2, was equal or better than the controlled rate freezing method. Our study implied that the combined use of cryoprotective agent CellBanker 2 and uncontrolled rate freezing could be a convenient cryopreservation method for peripheral blood mononuclear cells.

Recovery and maintenance of NESTIN expression in umbilical cord-MSC using a novel culture medium.

Mesenchymal stem cells (MSC) are a popular candidate in cellular therapy for many diseases. MSCs are well known by their feature of self-renewal and their differentiation potential. NESTIN is a cytoskeletal protein expressed in MSC that functions directly in cell proliferation and differentiation. Here, we demonstrated that adding UltraGRO, a medium supplement, could maintain and partially recover the expression of NESTIN in human umbilical cord derived MSCs (UC-MSCs). Furthermore, the UC-MSCs cultured with UltraGRO showed a better immunomodulation ability in a colitis mouse model compared with those cultured in other types of media. This indicates that the use of novel culture medium benefits the maintenance of NESTIN expression and NESTIN may be one of the vital factors that regulates the performance of MSCs.

Global crotonylome reveals CDYL-regulated RPA1 crotonylation in homologous recombination-mediated DNA repair.

Previously, we reported that chromodomain Y-like (CDYL) acts as a crotonyl-coenzyme A hydratase and negatively regulates histone crotonylation (Kcr). However, the global CDYL-regulated crotonylome remains unclear. Here, we report a large-scale proteomics analysis for protein Kcr. We identify 14,311 Kcr sites across 3734 proteins in HeLa cells, providing by far the largest crotonylome dataset. We show that depletion of CDYL alters crotonylome landscape affecting diverse cellular pathways. Specifically, CDYL negatively regulated Kcr of RPA1, and mutation of the Kcr sites of RPA1 impaired its interaction with single-stranded DNA and/or with components of resection machinery, supporting a key role of RPA1 Kcr in homologous recombination DNA repair. Together, our study indicates that protein crotonylation has important implication in various pathophysiological processes.

Polarized second-harmonic generation optical microscopy for laser-directed assembly of ZnO nanowires.

Two-photon polymerization (TPP) based on laser direct writing is currently one of the most prevailing 3D micro/nano fabrication techniques. Nanomaterials can be doped in resins and assembled by TPP for developing advanced 3D functional devices. However, there lacks an effective visualization tool to determine the distribution and orientation of the nanomaterials as-doped in the composite resins. Herein, we present a nondestructive, in situ, and rapid characterization method to determine the orientation and distribution of the nanomaterials within cured resins using polarized second-harmonic generation (p-SHG). The directional assembly of the ZnO nanowires within micro/nanostructures fabricated by TPP is, for the first time to the best of our knowledge, characterized by p-SHG optical microscopy with a fast imaging speed by two orders of magnitude higher than that of the Raman mapping technique. Our method opens a window for nondestructive, rapid, in situ, and polarization-resolved characterization of functional devices made by TPP micro/nanofabrication.

Recovery and functionality of cryopreserved peripheral blood mononuclear cells using five different xeno-free cryoprotective solutions.

In this study, we compared three commercially available and two widely used CPAs for their ability of cryopreserving PBMCs. Similar survival (81.0%) and recovery rate (73.7%) were observed among cells using these five CPAs. However, all the cryopreserved PBMCs exhibited a significantly lower survival rate when compared with the fresh samples (94.3%). We further evaluated effector cell subpopulation and tumoricidal activity of PBMC-derived cytokine-induced killing (CIK) cells and natural killing (NK) cells. Similar and high survival (CIK: 88.6%; NK: 87.5%) and recovery (CIK: 99.5%; NK: 99.7%) rates were detected in CIK and NK cells prepared from cryopreserved PBMCs using the five CPAs. The CD3+CD56+ effector percentage (27.3%) of cryopreserved PBMC-derived CIK cells using the five different CPAs and their tumoricidal activities on melanoma CHL-1 cells (45.7%) and bladder cancer cell line T-24 (44.7%) were similar but significantly lower than those of the fresh PBMC-derived controls (effector: 30.7%; CHL-1: 84.2%; T-24: 82.2%). Cryopreserved PBMC-derived NK cells also exhibited similar tumoricidal activities (CHL-1: 73.8%; T-24: 71.9%) but was significantly lower than that of the fresh control group. We were not able to identify a specific CPA that performed superior than others in PBMC cryopreservation.

Spinal trauma in mainland China from 2001 to 2007: an epidemiological study based on a nationwide database.

STUDY DESIGN: Descriptive epidemiological study. OBJECTIVE: To determine the epidemiological characteristics of spinal trauma in Mainland China. SUMMARY OF BACKGROUND DATA: To date, a large-scale epidemiological analysis of spinal trauma in Mainland China has not been undertaken. METHODS: Data were acquired from Chinese Database of Traumas. Patients with International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) coding of 805.x and 806.x (spinal column fracture with and without spinal cord injury) from 2001 to 2007 were identified. Variables assessed included patient demographics, etiology, segmental distribution, and outcomes. RESULTS: A total of 82,720 patients with spinal trauma were identified, accounting for 4.58% of all trauma patients in the study period. Spinal cord injuries (SCIs) and fracture-dislocations accounted for 16.87% and 7.17% of spinal trauma and 0.74% and 0.32% of all traumas in the same period, respectively. The male-to-female ratio was 2.33:1. About 79.32% of spinal trauma occurred in patients between 20 and 60 years of age. There was an annual increase in incidence during the study period. A total of 64,630 patients (78.13%) had a definitive cause, with motor vehicle accidents identified as the leading etiological factor (33.61%), followed by high falls (31.25%) and trivial falls (23.23%). Lumbar spine was most frequently involved (56.09%), followed by thoracic spine (23.77%), cervical spine (17.75%), and sacrococcygeal vertebrae (2.39%). Fracture-dislocation occurred most frequently in the cervical and lumbar spines, whereas spinal cord injury occurred most frequently in the cervical and thoracic spines. Children younger than 10 years of age were prone to cervical spine injury, whereas adults older than 60 years were more prone to osteoporotic thoracic and lumbar fractures. Overall rate of conservative treatment (55.88%) was higher than that of operative treatment (44.12%). Overall combined cure and improvement rates exceeded 90%. The male mortality rate was twice that of the female population. Lumbar spine injury was the most curable, whereas cervical spine injury was associated with the worst prognosis and the highest medical costs. CONCLUSION: This is the first large-scale epidemiological study of spinal injury in Mainland China. The results obtained have important implications for future public health care planning, public safety, and resource allocation.

The epidemiology of civilian inpatients' burns in Chinese military hospitals, 2001-2007.

BACKGROUND: In China, large sample research directed at the epidemiology of burns is still scarce. Given the leading position of Chinese military hospitals in domestic clinical practice pertaining to burns, a look into their cases would help shed light on the incidence and hospital care of burns in China today. METHODS: Data were collected from information systems of all the Chinese military hospitals, for the years 2001-2007 inclusive. Retrospective analyses on the epidemiology characteristics of burns from those in patients admitted to all the Chinese military hospitals within this period were conducted. RESULTS: A total of 172,256 cases were studied, with a total number of 1384 fatalities, and the median length of hospital stay (LOS) 11.00 days (interquartile range 6.00-19.00 days). Rate of burn incidence remained high in May, June, July and August. Children of preschool aged and adults in the labor force were at the highest risk of injury. Scalds constituted the majority of injuries (80.5%). Multiple body site burns are the most commonly seen category (31.98%) in terms of injured body region distribution. In fire-related burns, major burns (TBSA>30%) represented a significant proportion. Males with TBSA>30%, fire-related burns resulted in the greatest number of fatalities. Gender, age, site of injury, burn size and outcome are all correlated with the length of hospitalization. CONCLUSION: Although this paper cannot provide information on population estimates of burns due to the absence of data on burns attending civilian hospitals, it finds that the epidemiology of burns in China shows distinct characteristics compared with previous studies of other country and regions, including that of other developing or low and middle-income countries (LMICs), irrespective of China's inclusion in that group.