Micro-flow cell washing technique combined with single-cell Raman spectroscopy for rapid and automatic antimicrobial susceptibility test of pathogen in urine.

Facing the rapid spread of antimicrobial resistance, methods based on single-cell Raman spectroscopy have proven their advances in reducing the turn-around time (TAT) of antimicrobial susceptibility tests (AST). However, the Raman-based methods are still hindered by the prolonged centrifugal cell washing procedure, which may require complex labor operation and induce high mechanical stress, resulting in a pretreatment time of over 1 h as well as a high cell-loss probability. In this study, we developed a micro-flow cell washing device and corresponding Raman-compatible washing chips, which were able to automatically remove the impurities in the samples, retain the bacterial cell and perform Raman spectra acquisition in situ. Results of washing the 5- and 10-µm polymethyl methacrylate (PMMA) microspheres showed that the novel technique achieved a successful removal of 99 % impurity and an 80 % particle retention rate after 6 to 10 cycles of washing. The micro-flow cell washing technique could complete the pretreatment for urine samples in a 96-well plate within 10 min, only taking 15 % of the handling time required by centrifugation. The AST profiles of urine sample spiked with E. coli 25922, E. faecalis 29212, and S. aureus 29213 obtained by the proposed Raman-based approach were found to be 100 % consistent with the results from broth micro-dilution while reducing the TAT to 3 h from several days which is required by the latter. Our study has demonstrated the micro-flow cell washing technique is a reliable, fast and compatible approach to replace centrifuge washing for sample pretreatment of Raman-AST and could be readily applied in clinical scenarios.

Disturbed glycolipid metabolism activates CXCL13-CXCR5 axis in senescent TSCs to promote heterotopic ossification.

Heterotopic ossification (HO) occurs as a common complication after injury, while its risk factor and mechanism remain unclear, which restricts the development of pharmacological treatment. Clinical research suggests that diabetes mellitus (DM) patients are prone to developing HO in the tendon, but solid evidence and mechanical research are still needed. Here, we combined the clinical samples and the DM mice model to identify that disordered glycolipid metabolism aggravates the senescence of tendon-derived stem cells (TSCs) and promotes osteogenic differentiation. Then, combining the RNA-seq results of the aging tendon, we detected the abnormally activated autocrine CXCL13-CXCR5 axis in TSCs cultured in a high fat, high glucose (HFHG) environment and also in the aged tendon. Genetic inhibition of CXCL13 successfully alleviated HO formation in DM mice, providing a potential therapeutic target for suppressing HO formation in DM patients after trauma or surgery.

A Prospective Self-controlled Clinical Trial of Nonactivated Low Leukocyte PRP in Female Pattern Hair Loss Patients of Childbearing Age.

BACKGROUND: Alopecia significantly affects the mental health and social relationship of women since childbearing age, highlighting the need for a safe, effective, and convenient treatment. METHODS: The authors have conducted a prospective self-controlled trial involving 15 female patients at childbearing age with alopecia. These patients received a subcutaneous scalp injection of platelet-rich plasma once every 4 weeks for 3 treatments in total. Outcome measurements were included below: changes in hair density (hair/cm2), hair follicle density (hair follicle/cm2), and overall photographic assessment (improved or not) at 4, 12, and 24 weeks right after the first treatment. RESULTS: Comparing the photographs taken before and after the intervention, 67% of patients' hair density increased from 151 ± 39.82 hairs/cm2 (preintervention) to 170.96 ± 37.14 hairs/cm2 (at 24-week follow-up), representing an approximate increase of 19 hairs/cm2. Meanwhile, hair follicle density increased by approximately 15 follicles/cm2 after 24 weeks since the first treatment, rising from 151.04 ± 41.99 follicles/cm2 to 166.72 ± 37.13 follicles/cm2. The primary adverse reactions observed were local swelling and pain due to injections. CONCLUSION: Local injection of nonactivated platelet-rich plasma with low leukocytes concentration could be an effective strategy to alleviate alopecia symptoms in female patients.

Simulating PDT of port-wine stains in the in vivo chicken wattle model using Hemoporfin and radiation at 532 nm: Comparison of a LED and a laser source.

Port-wine stain (PWS) birthmarks are congenital capillary malformations occurring in 0.3 %∼0.5 % of newborns. Hemoporfin-mediated vascular-acting photodynamic therapy (Hemoporfin PDT) is an emerging option for treating PWS. This in vivo study aimed to compare laser and light-emitting diodes (LED) as light source for Hemoporfin PDT. Chicken wattles were used as the animal model. Color and histopathological changes were evaluated after combining Hemoporfin with KTP laser or LED light source of 532 nm at the same doses. Both PDT approaches could induce significant vascular injury and color bleaching. Although the use of the laser resulted in a greater vascular clearance, the LED showed more uniform distribution both in the beam profiles and tissue reaction and exhibited better safety. This in vivo study suggests that the LED is a favorable choice for larger PWS lesion.

Microwave-responsive gadolinium metal-organic frameworks nanosystem for MRI-guided cancer thermotherapy and synergistic immunotherapy.

The clinical application of cancer immunotherapy is unsatisfied due to low response rates and systemic immune-related adverse events. Microwave hyperthermia can be used as a synergistic immunotherapy to amplify the antitumor effect. Herein, we designed a Gd-based metal-organic framework (Gd-MOF) nanosystem for MRI-guided thermotherapy and synergistic immunotherapy, which featured high performance in drug loading and tumor tissue penetration. The PD-1 inhibitor (aPD-1) was initially loaded in the porous Gd-MOF (Gd/M) nanosystem. Then, the phase change material (PCM) and the cancer cell membrane were further sequentially modified on the surface of Gd/MP to obtain Gd-MOF@aPD-1@CM (Gd/MPC). When entering the tumor microenvironment (TME), Gd/MPC induces immunogenic death of tumor cells through microwave thermal responsiveness, improves tumor suppressive immune microenvironment and further enhances anti-tumor ability of T cells by releasing aPD-1. Meanwhile, Gd/MPC can be used for contrast-enhanced MRI. Transcriptomics data revealed that the downregulation of MSK2 in cancer cells leads to the downregulation of c-fos and c-jun, and ultimately leads to the apoptosis of cancer cells after treatment. In general, Gd/MPC nanosystem not only solves the problem of system side effect, but also achieves the controlled drug release via PCM, providing a promising theranostic nanoplatform for development of cancer combination immunotherapy.

1064nm Nd:YAG laser promotes chondrocytes regeneration and cartilage reshaping by upregulating local estrogen levels.

Cartilage is frequently used as a scaffolds for repairing and reconstructing body surface organs. However, after successful plastic surgery, transplanted cartilage scaffolds often exhibit deformation and absorption over time. To enhance the shaping stability of cartilage scaffolds and improve patients' satisfaction after reconstructions, we employed the ear folding models in New Zealand rabbits to confirm whether the 1064nm neodymium-doped yttrium aluminum garnet (Nd:YAG) laser could promote cartilage reshaping. There was an increase in collagen and aromatase (Cyp19) expression within the ear cartilage after laser treatment. Moreover, we have found that the Cyp19 inhibitor can inhibit the laser's effect on cartilage shaping and reduce collagen and Cyp19 expression. The overall findings suggest that treatment with 1064nm Nd:YAG laser irradiation can enhance estrogen levels in local cartilage tissues by upregulating Cyp19 expression in chondrocytes through photobiomodulation, thereby promoting the proliferation and collagen secretion of chondrocytes to improve cartilage reshaping and stability.

An innovative virtual reality training tool for the pre-hospital treatment of cranialmaxillofacial trauma.

Virtual reality (VR) surgery using the High Technology Computer Corporation Very Immersive Virtual Experience professional 2(HTC VIVE Pro2) suite is a multi-sensory, holistic surgical training experience. A multimedia combination including videos and three-dimensional interaction in VR has been developed to enable trainees to experience a realistic battlefield environment. The innovation allows trainees to interact with the individual components of the cranialmaxillofacial(CMF) anatomy and apply surgical instruments while watching close-up stereoscopic three-dimensional videos of the surgery. In this study, a novel training tool for the pre-hospital treatment of CMF trauma based on immersive virtual reality (iVR) was developed and validated. Twenty-five CMF surgeons evaluated the application for face and content validity. Using a structured assessment process, the surgeons commented on the content of the developed training tool, its realism and usability and the applicability of VR surgery for CMF trauma rescue simulation training. The results confirmed the applicability of VR for delivering training in the pre-hospital treatment of CMF trauma. Modifications were suggested to improve the user experience and interactions with the surgical instruments. This training tool is ready for testing with surgical trainees.

Study on Application of YAG Laser-Assisted Ear Correction Model to Congenital Auricular Deformities.

BACKGROUND: The objective of this study was to compare the clinical effects of a long-pulse-width 1064-nm Nd:YAG laser-assisted ear correction model and a simple ear correction model in the treatment of congenital auricular deformities in children older than 6 weeks. METHODS: A total of 129 children (179 ears) with congenital auricular deformities who were older than 2 months were included. Among them, 84 (119 ears) were treated with the ear correction model after long-pulse-width 1064-nm Nd:YAG laser irradiation (observation group) and 45 (60 ears) were treated with the ear correction model only (control group). The affected ears of the two groups were measured, the correction time was calculated, and the patients' family members filled in the satisfaction questionnaire before and after the treatment. RESULTS: The level of satisfaction of both groups improved after the treatment. The satisfaction level of the observation group was higher than that of the control group. Fewer days of correction were observed in the observation group than in the control group ( p < 0.05). The measurements of the affected ears in the two groups were significantly different after treatment. The difference after treatment of each measurement in the observation group was higher than in the control group. The therapeutic effect of the observation group was significantly better than that of the control group and the differences were statistically significant. CONCLUSION: Long-pulse-width 1064-nm Nd:YAG laser combined with the ear correction model can improve the success rate of correction of congenital auricular deformities in over-aged ears. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, II.

Objective Assessment of the Effectiveness of the Transconjunctival Fat-Repositioning Technique Using Magnetic Resonance Imaging and Three-Dimensional Surface Imaging.

Background: To quantify the surgical outcomes of transconjunctival fat repositioning technique, we study the lower periorbital area with magnetic resonance imaging (MRI) and three-dimensional (3D) surface imaging. Methods: The preoperative and postoperative images were analyzed. The T1-weighted MR images in the quasi-sagittal plane parallel to the long axis of the orbit and passing through the globe center were used to measure the transposed fat, and 3D comparison was performed to calculate the tear trough depth. Changes throughout the surgery and their relationships with clinical characteristics were statistically analyzed. Results: A total of 11 patients who underwent surgery were included. The mean follow-up period was 10.1 months. MRI measurements showed that the fat area beneath the tear trough increased from 0 to 22.7 ± 6.7 mm2 after fat repositioning, whereas the fat area of the eyelid bag decreased from 46.3 ± 11.0 mm2 to 25.5 ± 14.0 mm2. The 3D results revealed an increase in thickness by 1.2 ± 0.4 mm in the tear tough areas. Conclusion: Both MRI and 3D imaging provide options to objectively assess the fat-repositioning technique.

Accelerated Skin Wound Healing Using Flexible Photovoltaic-Bioelectrode Electrical Stimulation.

Owing to the complex and long-term treatment of foot wounds due to diabetes and the limited mobility of patients, advanced clinical surgery often uses wearable flexible devices for auxiliary treatment. Therefore, there is an urgent need for self-powered biomedical devices to reduce the extra weight. We have prepared an electrically stimulated MEMS (Micro Electromechanical System) electrode integrated with wearable OPV (Organic photovoltaic). The wearable OPV is constructed of a bio-affinity PET-ITO substrate and a hundred-nanometer organic layer. Under sunlight and near-infrared light irradiation, a voltage and current are supplied to the MEMS electrode to generate an exogenous lateral electric field directed to the center of the wound. The results of in vitro cell experiments and diabetic skin-relieving biological experiments showed the proliferation of skin fibroblasts and the expression of transforming growth factors increased, and the skin wounds of diabetic mouse healed faster. Our research provides new insights for the clinical treatment of diabetes.

High-quality and large-area echelle grating ruled on 500-mm ruling engine for the fiber array solar optical telescope (FASOT).

A 300 mm×500 mm large-area echelle grating with groove density of 79 grooves/mm is fabricated for the spectrometer of the fiber array solar optical telescope (FASOT). This paper focusses on measurement methods of the grating performance. We present a method to evaluate the grating's stray light intensity, which is measured to a level of 10-4. The directly measured grating efficiency is approximately 90% of the designed value, and an indirect measurement method based on the grating groove profile is proposed. Based on the Rayleigh criterion and the grating diffraction wavefront, a physical optics method and a geometric grating method are proposed and are used to calculate the actual grating resolving power; the calculated results exceed 95% of the grating's theoretical resolving power. These results show that the CIOMP-6 ruling engine has sufficient precision to fabricate high-quality, large-area echelle gratings.

Effect of the measuring mirror surface shape error on ruled groove straightness and the grating diffraction wavefront.

Measuring mirror requirements and their impact on groove errors are related to the error compensation strategy for a ruling engine. We analyze why the measuring mirror of the CIOMP-6 engine affects the groove straightness and the grating diffraction wavefront. We study a theoretical model of the relationship between the measuring mirror's surface shape error and the grating wavefront, propose a requirement for the measuring mirror surface shape error, and reprocess the measuring mirror. Comparative ruling experiments prove that the grating's wavefront quality at the diffraction order along the groove direction improved significantly after reprocessing of the measuring mirror.

Numerical calculation of the mosaic error between mosaic gratings.

The theoretical calculation model for a mosaic error was established based on the plane equation for a grating surface and the relationship equation for a mosaic grating surface. A mosaic grating was obtained based on this model. In the experiment, the mosaic error was calculated based on the diffraction wavefronts of two groups of mosaic gratings that were obtained simultaneously with a Zygo interferometer. The difference between the wavefront of the mosaic grating and the average wavefront of the mosaic grating element was 0.031λ. The maximum far-field intensity of the mosaic grating was 90% of that without an error. This model provides a theoretical basis for the numerical mosaic between gratings. In addition, the mosaic error can be calculated with this model, and the quality of the mosaic grating can be evaluated.

Death Caused by Vaginal Injection of Hyaluronic Acid and Collagen: A Case Report.

With the expanding utilization of hyaluronic acid (HA) and collagen as cosmetic fillers in plastic and reconstructive surgery, complications due to their excessive use and/or irregular procedures warrant great caution. Recently, a fatal case occurred caused by a poorly regulated procedure of vaginal injection of HA and collagen. A 33-year-old female was admitted to the emergency department 3 hours after the operation with a chief complaint of dyspnea, which initiated 5 to 10 minutes after the operation. Her blood pressure remained low while dopamine pressor and fluid replacement were used. Computed tomography of the chest showed local exudation in the lower lobe of the left lung, enlargement of right atrium and ventricle, and uneven development of the bilateral inferior lobar artery with filling defects. Pulmonary computed tomography angiography and three-dimensional reconstruction showed continuous interruption of pulmonary artery branches of the posterior basal segment of the right lower lobe. Unfortunately, the clinical symptoms caused by vaginal injection aggravated rapidly and could not be effectively controlled. The patient died 9 hours after injection. Pulmonary complications after injection of cosmetic fillers are scarcely reported. Thus far, only 2 cases of HA-related pulmonary complications after vaginal injection have been described. The present case emphasizes that surgeons and other healthcare providers must be aware of the risk of serious pulmonary complications and even death associated with these 2 widely utilized injectable fillers. Level of Evidence: 5.

Ruling engine using adjustable diamond and interferometric control for high-quality gratings and large echelles.

A concept for position adjustment of the diamond in a ruling engine to enable nanoscale groove positioning is proposed. Based on this concept, we fabricate a diamond carriage, design an optical path, and propose an appropriate control method. Several high-quality gratings are ruled for spectrometer or interferometer applications. After implementation of the improvements, the maximum intensity of ghosts and scattered light of the echelle grating with 79 grooves/mm is reduced to half of that before the improvement, and the highest achievable groove density is increased from 6000 grooves/mm to 8000 grooves/mm. The grating ruling results indicate that the proposed concept and the related improvements to the engine significantly improve the accuracy of the CIOMP-6 ruling engine.

Method for calculation and analysis of the weights of grating mosaic errors.

A method to calculate the weight of grating mosaic errors is proposed based on an analytical hierarchy process. An accurate mosaic error tolerance calculation formula is also presented that is useful for large-size grating fabrication by the mosaic method. The grating mosaic error weights and tolerances are analyzed for mosaic echelle gratings. The analytical error weight and tolerance results agree well with the experimental results. The mosaic grating's far-field intensity is 95.8%, which is higher than the 94% value from the simulation results.

The deubiquitylating enzyme USP15 regulates homologous recombination repair and cancer cell response to PARP inhibitors.

Poly-(ADP-ribose) polymerase inhibitors (PARPi) selectively kill breast and ovarian cancers with defects in homologous recombination (HR) caused by BRCA1/2 mutations. There is also clinical evidence for the utility of PARPi in breast and ovarian cancers without BRCA mutations, but the underlying mechanism is not clear. Here, we report that the deubiquitylating enzyme USP15 affects cancer cell response to PARPi by regulating HR. Mechanistically, USP15 is recruited to DNA double-strand breaks (DSBs) by MDC1, which requires the FHA domain of MDC1 and phosphorylated Ser678 of USP15. Subsequently, USP15 deubiquitinates BARD1 BRCT domain, and promotes BARD1-HP1γ interaction, resulting in BRCA1/BARD1 retention at DSBs. USP15 knockout mice exhibit genomic instability in vivo. Furthermore, cancer-associated USP15 mutations, with decreased USP15-BARD1 interaction, increases PARP inhibitor sensitivity in cancer cells. Thus, our results identify a novel regulator of HR, which is a potential biomarker for therapeutic treatment using PARP inhibitors in cancers.

Analysis and removal of five-dimensional mosaicking errors in mosaic grating.

Combining the mathematical relationships between the grating wavefront and surfaces with the spatial relationships between the two grating wavefront, a mathematical model of the mosaicking errors is established to mosaic gratings. The five-dimensional mosaicking errors will respectively be calculated and then removed by the adjustment mechanisms. Mosaicking experiments are performed by using two gratings. First, by using zeroth order, the longitudinal offset is calculated and removed. Second, by using the diffraction order, the in-plane angle and grating spacing are calculated and removed, while the tip and tilt angles are calculated and removed. And then a mosaic grating is obtained.

Broadband, high-resolution Raman observations from a double-echelle spatial heterodyne Raman spectrometer.

A new broadband Raman spectrometer has been developed, to the best of our knowledge, using a double-echelle spatial heterodyne Raman spectrometer (DESHRS). The instrument is constructed by using two echelle gratings. Masks are used to remove the shadow ghosts caused by the different orders of the two echelle gratings. Raman spectra of inorganic solid targets and methanol are given, and Raman shifts of up to 3000 cm-1 are obtained by the DESHRS. The instrument has shown that a broadband coverage and high resolution can be achieved simultaneously to meet the requirements of Raman measurements, covering 3590 cm-1 with 1.21 cm-1 spectral resolution.

Backscattering Raman spectroscopy using multi-grating spatial heterodyne Raman spectrometer.

Spatial heterodyne Raman spectrometry (SHRS) is a spectral analysis technique used to study material structures and compositions. We propose a multi-grating SHRS system that uses a multi-grating module rather than the single grating used to terminate each arm in traditional spatial heterodyne spectrometry (SHS). The proposed system not only retains the advantages of traditional SHS but also resolves the mutual limitation between system spectral range and resolution. The increased spectral range and resolution that can be achieved in detection are dependent on the number of sub-gratings used in the module. A verification system was built using 130 gr/mm and 150 gr/mm sub-gratings and calibrated. Under different experimental conditions (including laser power, integration time, container material and thickness, pure and mixed samples, and standoff experiments), the backscattered Raman spectra of different types of targets (including organic solutions, inorganic powders, and minerals) were tested. The multi-grating SHRS shows good performance for broad spectral range and high-resolution Raman detection.