Preparation of (S)-epichlorohydrin using a novel halohydrin dehalogenase by selective conformation adjustment.

Chiral epichlorohydrin (ECH) is an attractive intermediate for chiral pharmaceuticals and chemicals preparation. The asymmetric synthesis of chiral ECH using 1,3-dicholoro-2-propanol (1,3-DCP) catalyzed by a haloalcohol dehalogenase (HHDH) was considered as a feasible approach. However, the reverse ring opening reaction caused low optical purity of chiral ECH, thus severely restricts the industrial application of HHDHs. In the present study, a novel selective conformation adjustment strategy was developed with an engineered HheCPS to regulate the kinetic parameters of the forward and reverse reactions, based on site saturation mutation and molecular simulation analysis. The HheCPS mutant E85P was constructed with a markable change in the conformation of (S)-ECH in the substrate pocket and a slight impact on the interaction between 1,3-DCP and the enzyme, which resulted in the kinetic deceleration of the reverse reactions. Compared with HheCPS, the catalytic efficiency (kcat(S)-ECH/Km(S)-ECH) of the reversed reaction dropped to 0.23-fold (from 0.13 to 0.03 mM-1 s-1), while the catalytic efficiency (kcat(1,3-DCP)/Km(1,3-DCP)) of the forward reaction only reduced from 0.83 to 0.71 mM-1 s-1. With 40 mM 1,3-DCP as substrate, HheCPS E85P catalyzed the synthesis of (S)-ECH with the yield up to 55.35% and the e.e. increased from 92.54 to >99%. Our work provided an effective approach for understanding the stereoselective catalytic mechanism as well as the green manufacturing of chiral epoxides.

Precision off-the-shelf natural killer cell therapies for oncology with logic-gated gene circuits.

Acute myeloid leukemia (AML) is an aggressive disease with a poor prognosis (5-year survival rate of 30.5% in the United States). Designing cell therapies to target AML is challenging because no single tumor-associated antigen (TAA) is highly expressed on all cancer subpopulations. Furthermore, TAAs are also expressed on healthy cells, leading to toxicity risk. To address these targeting challenges, we engineer natural killer (NK) cells with a multi-input gene circuit consisting of chimeric antigen receptors (CARs) controlled by OR and NOT logic gates. The OR gate kills a range of AML cells from leukemic stem cells to blasts using a bivalent CAR targeting FLT3 and/or CD33. The NOT gate protects healthy hematopoietic stem cells (HSCs) using an inhibitory CAR targeting endomucin, a protective antigen unique to healthy HSCs. NK cells with the combined OR-NOT gene circuit kill multiple AML subtypes and protect primary HSCs, and the circuit also works in vivo.

Development of a risk prediction model for subsequent infection after colonization with carbapenem-resistant Enterobacterales: a retrospective cohort study.

BACKGROUND: Colonization of carbapenem-resistant Enterobacterale (CRE) is considered as one of vital preconditions for infection, with corresponding high morbidity and mortality. It is important to construct a reliable prediction model for those CRE carriers with high risk of infection. METHODS: A retrospective cohort study was conducted in two Chinese tertiary hospitals for patients with CRE colonization from 2011 to 2021. Univariable analysis and the Fine-Gray sub-distribution hazard model were utilized to identify potential predictors for CRE-colonized infection, while death was the competing event. A nomogram was established to predict 30-day and 60-day risk of CRE-colonized infection. RESULTS: 879 eligible patients were enrolled in our study and divided into training (n = 761) and validation (n = 118) group, respectively. There were 196 (25.8%) patients suffered from subsequent CRE infection. The median duration of subsequent infection after identification of CRE colonization was 20 (interquartile range [IQR], 14-32) days. Multisite colonization, polymicrobial colonization, catheterization and receiving albumin after colonization, concomitant respiratory diseases, receiving carbapenems and antimicrobial combination therapy before CRE colonization within 90 days were included in final model. Model discrimination and calibration were acceptable for predicting the probability of 60-day CRE-colonized infection in both training (area under the curve [AUC], 74.7) and validation dataset (AUC, 81.1). Decision-curve analysis revealed a significantly better net benefit in current model. Our prediction model is freely available online at https://ken-zheng.shinyapps.io/PredictingModelofCREcolonizedInfection/ . CONCLUSIONS: Our nomogram has a good predictive performance and could contribute to early identification of CRE carriers with a high-risk of subsequent infection, although external validation would be required.

Lymph node dissection in small-sized pulmonary metastasectomy: Impact on the long-term survival.

INTRODUCTION: Pulmonary metastasectomy has been clarified in improving long-term survival in most primary malignancies with pulmonary metastasis, while the role of additional lymph node dissection remained controversial. We aimed to investigate the prognosis of lymph node involvement and identify the role of lymph node dissection during pulmonary metastasectomy in a real-world cohort. METHODS: We identified patients diagnosed with pulmonary metastases with ≤3 cm in size and received pulmonary metastasectomy between 2004 and 2017 in the Surveillance, Epidemiology, and End Results database. We compared the survival via Kaplan-Meier analysis and propensity score matching method, and the multivariable analysis was conducted by cox regression analysis. RESULTS: A total of 3452 patients were included, of which 2268(65.7%) received lymph node dissection, and the incidence of node-positive was 11.3%(256/2268). In total, the median overall survival was 62.8 months(interquartile range, 28.6-118.9 months), and the lymph node involvement was referred to an impaired survival compared to node-negative diseases(5-year overall survival rate, 58.0% versus 38.6%), with comparable survival between N1 and N2 diseases(P = 0.774). Lymph node dissection was associated with improved survival(HR = 0.80; 95%CI, 0.71-0.90; P < 0.001), and the survival benefits remained regardless of age, sex, the number of metastases, and surgical procedures, even in those with node-negative diseases. At least eight LNDs might lead to a significant improvement in survival, and additional survival benefits might be limited with additional dissected lymph nodes. CONCLUSIONS: Lymph node involvement was associated with impaired survival, and lymph node dissection during pulmonary metastasectomy could improve long-term survival and more accurate staging.

Acute kidney injury should not be neglected - optimization of quick Pitt bacteremia score for predicting mortality in critically ill patients with bloodstream infection: a retrospective cohort study.

Background: Considering the therapeutic difficulties and mortality associated with bloodstream infection (BSI), it is essential to investigate other potential factors affecting mortality in critically ill patients with BSI and examine the utility of the quick Pitt bacteremia (qPitt) score to improve the survival rate. Objectives: To improve the predictive accuracy of the qPitt scoring system by evaluating the five current components of qPitt and including other potential factors influencing mortality in critically ill patients with BSI. Design: This was a retrospective cohort study. Methods: Medical information from the Medical Information Mart for Intensive Care IV database was used in this retrospective cohort study. The risk factors associated with mortality were examined using a multivariate logistic regression model. The area under the receiver operating characteristic curve (AUC) was used to assess the discriminatory capability of the prediction models. Results: In total, 1240 eligible critically ill patients with BSI were included. After adjustment for age, community-onset BSI, indwelling invasive lines, and Glasgow Coma Scale (GCS) ⩽ 8, acute kidney injury (AKI) was identified as a notable risk factor for 14-day mortality. Except for altered mental status, the four other main components of the original qPitt were significantly associated with 14-day mortality. Hence, we established a modified qPitt (m-qPitt) by adding AKI and replacing altered mental status with GCS ⩽ 8. The AUCs for m-qPitt and qPitt were 0.723 [95% confidence interval (CI): 0.683-0.759] and 0.708 (95% CI: 0.669-0.745) in predicting 14-day mortality, respectively. Moreover, m-qPitt also had acceptable performance and discrimination power [0.700 (95% CI: 0.666-0.732)] in predicting 28-day mortality. Conclusion: AKI significantly influenced the survival of critically ill patients with BSIs. Compared with the original qPitt, our new m-qPitt was proven to have a better predictive performance for mortality in critically ill patients with BSI. Further studies should be conducted to validate the practicality of m-qPitt.

En bloc resection of an extremely giant mediastinal immature teratoma with somatic-type malignancy: A case report with a brief review of the literature.

Primary mediastinum immature teratoma with somatic-type malignant transformation (SM) is extremely rare, and the clinical prognosis is poor. Immature teratoma with SM is difficult to eradicate by chemotherapy due to poor sensitivity; therefore, surgical resection is recommended whenever possible because it may offer better survival.

Non-IL-2-blocking anti-CD25 antibody inhibits tumor growth by depleting Tregs and has synergistic effects with anti-CTLA-4 therapy.

CD25, also known as the interleukin-2 receptor α chain (IL-2Rα), is highly expressed on regulatory T cells (Tregs), but relatively lower on effector T cells (Teffs). This makes it a potential target for Treg depletion, which can be used in tumor immunotherapy. However, marketed anti-CD25 antibodies (Basiliximab and Daclizumab) were originally developed as immunosuppressive drugs to prevent graft rejection, because these antibodies can block IL-2 binding to CD25 on Teffs, which in turn destroys the function of Teffs. Recent studies have shown that non-IL-2-blocking anti-CD25 antibodies have displayed exciting antitumor effects. Here, we screened out a non-IL-2-blocking anti-CD25 monoclonal antibody (mAb) 7B7 by hybridoma technology, and confirmed its antitumor activity via depleting Tregs in a CD25 humanized mouse model. Subsequently, we verified that the humanized 7B7, named as h7B7-15S, has comparable activities to 7B7, and that its Treg depletion is further increased when combined with anti-CTLA-4, leading to enhanced remodeling of the tumor immune microenvironment. Moreover, our findings reveal that the Fab form of h7B7-15S has the ability to deplete Tregs, independent of the Fc region. Taken together, our studies expand the application of anti-CD25 in tumor immunotherapy and provide insight into the underlying mechanism.

Human Movement Recognition Based on 3D Point Cloud Spatiotemporal Information from Millimeter-Wave Radar.

Human movement recognition is the use of perceptual technology to collect some of the limb or body movements presented. This practice involves the use of wireless signals, processing, and classification to identify some of the regular movements of the human body. It has a wide range of application prospects, including in intelligent pensions, remote health monitoring, and child supervision. Among the traditional human movement recognition methods, the widely used ones are video image-based recognition technology and Wi-Fi-based recognition technology. However, in some dim and imperfect weather environments, it is not easy to maintain a high performance and recognition rate for human movement recognition using video images. There is the problem of a low recognition degree for Wi-Fi recognition of human movement in the case of a complex environment. Most of the previous research on human movement recognition is based on LiDAR perception technology. LiDAR scanning using a three-dimensional static point cloud can only present the point cloud characteristics of static objects; it struggles to reflect all the characteristics of moving objects. In addition, due to its consideration of privacy and security issues, the dynamic millimeter-wave radar point cloud used in the previous study on the existing problems of human body movement recognition performance is better, with the recognition of human movement characteristics in non-line-of-sight situations as well as better protection of people's privacy. In this paper, we propose a human motion feature recognition system (PNHM) based on spatiotemporal information of the 3D point cloud of millimeter-wave radar, design a neural network based on the network PointNet++ in order to effectively recognize human motion features, and study four human motions based on the threshold method. The data set of the four movements of the human body at two angles in two experimental environments was constructed. This paper compares four standard mainstream 3D point cloud human action recognition models for the system. The experimental results show that the recognition accuracy of the human body's when walking upright can reach 94%, the recognition accuracy when moving from squatting to standing can reach 84%, that when moving from standing to sitting can reach 87%, and the recognition accuracy of falling can reach 93%.

Long-Wavelength Illumination-Induced Photocurrent Enhancement of a ZnPc Photocathodic Material for Bioanalytical Applications.

Herein, a novel photocathodic nanocomposite poly{4,8-bis[5-(2-ethylhexyl)-thiophen-2-yl] benzo[1,2-b:4,5-b']dithiophene-2,6-diyl-alt-3-fluoro-2-[(2-ethylhexyl)-carbonyl]thieno[3,4-b]thiophene-4,6-diyl}/phthalocyanine zinc (PTB7-Th/ZnPc) with high photoelectric conversion efficiency under long-wavelength illumination was prepared to construct an ultrasensitive biosensor for the detection of microRNA-21 (miRNA-21), accompanied by a prominent anti-interference capability toward reductive substances. Impressively, the new heterojunction PTB7-Th/ZnPc nanocomposite could not only generate a strong cathodic photocurrent to improve the detection sensitivity under long-wavelength illumination (660 nm) but also effectively avoid the high damage of biological activity caused by short-wavelength light stimulation. Accordingly, by coupling with rolling circle amplification (RCA)-triggered DNA amplification to form functional biquencher nanospheres, a PEC biosensor was fabricated to realize the ultrasensitive analysis of miRNA-21 in the concentration range of 0.1 fM to 10 nM with a detection limit as low as 32 aM. This strategy provided a novel long-wavelength illumination-induced photocurrent enhancement photoactive material for a sensitive and low-damage anti-interference bioassay and early clinical disease diagnosis.

Effect of antivirals plus low-dose, short-term glucocorticoids on post-herpetic neuralgia.

BACKGROUND: Post-herpetic neuralgia (PHN) is the most common complication of herpes zoster (HZ). It is unclear whether short-term and low-dose glucocorticoids combined with antivirals can reduce the incidence of PHN. OBJECTIVES: To investigate the effects of antivirals plus low-dose, short-term glucocorticoids on PHN. MATERIALS & METHODS: A total of 394 patients with HZ were divided into glucocorticoid and non-glucocorticoid groups, and the incidence of PHN was studied retrospectively. Forty patients with HZ were randomized into the glucocorticoid (n=20) and non-glucocorticoid (n=20) groups. The levels of protein 100-B (S100B) and neuron-specific enolase (NSE) in the blood and the viral load in the skin lesions were measured before and after seven days of treatment. Patient condition and pain were assessed using the HZ and visual analogue scale pain scores. RESULTS: The incidence of PHN in the glucocorticoid and non-glucocorticoid groups was 20.89% and 30.51%, respectively. In patients with onset time >seven days before treatment, the incidence of PHN was 19.81% and 40.16%, respectively. In the glucocorticoid group, after treatment, the mean serum NSE level of the glucocorticoid group decreased from 15.8 ng/mL to 14.0 ng/mL, while the mean serum S100B level decreased from 1486.3 ng/mL to 1453.7 ng/mL. There was no intergroup difference in the reduction rate of viral load. The mean condition score and pain score were significantly lower in the glucocorticoid group. CONCLUSION: Antiviral therapy plus low-dose, short-term glucocorticoids can improve the condition of patients with HZ and partly reduce the incidence of PHN.

mm-TPG: Traffic Policemen Gesture Recognition Based on Millimeter Wave Radar Point Cloud.

Automatic driving technology refers to equipment such as vehicle-mounted sensors and computers that are used to navigate and control vehicles autonomously by acquiring external environmental information. To achieve automatic driving, vehicles must be able to perceive the surrounding environment and recognize and understand traffic signs, traffic signals, pedestrians, and other traffic participants, as well as accurately plan and control their path. Recognition of traffic signs and signals is an essential part of automatic driving technology, and gesture recognition is a crucial aspect of traffic-signal recognition. This article introduces mm-TPG, a traffic-police gesture recognition system based on a millimeter-wave point cloud. The system uses a 60 GHz frequency-modulated continuous-wave (FMCW) millimeter-wave radar as a sensor to achieve high-precision recognition of traffic-police gestures. Initially, a double-threshold filtering algorithm is used to denoise the millimeter-wave raw data, followed by multi-frame synthesis processing of the generated point cloud data and feature extraction using a ResNet18 network. Finally, gated recurrent units are used for classification to enable the recognition of different traffic-police gestures. Experimental results demonstrate that the mm-TPG system has high accuracy and robustness and can effectively recognize traffic-police gestures in complex environments such as varying lighting and weather conditions, providing strong support for traffic safety.

New hope for tumor immunotherapy: the macrophage-related "do not eat me" signaling pathway.

The "do not eat me" signaling pathway is extremely active in tumor cells, providing a means for these cells to elude macrophage phagocytosis and escape immune surveillance. Representative markers of this pathway, such as CD47 and CD24, are highly expressed in numerous tumors. The interaction of SIRPα with CD47 reduces the accumulation of non-myosin ⅡA on the cell membrane. The combination of CD24 and Siglec10 ultimately leads to the recruitment of SHP-1 or SHP-2 to reduce signal transduction. Both of them weaken the ability of macrophages to engulf tumor cells. Blocking the mutual recognition between CD47-SIRPα or CD24-Siglec10 using large molecular proteins or small molecular drugs represents a promising avenue for tumor immunotherapy. Doing so can inhibit signal transduction and enhance macrophage clearance rates of cancer cells. In this paper, we summarize the characteristics of the drugs that affect the "do not eat me" signaling pathway via classical large molecular proteins and small molecule drugs, which target the CD47-SIRPα and CD24-Siglec10 signaling pathways, which target the CD47-SIRPα and CD24-Siglec10 signaling pathways. We expect it will offer insight into the development of new drugs centered on blocking the "do not eat me" signaling pathway.

CRISPR-Mediated Base Editing: Promises and Challenges for a Viable Oncotherapy Strategy.

Base editing technology, developed from the CRISPR/Cas9 system, is able to efficiently implement single-base substitutions at specific DNA or RNA sites without generating double-strand breaks with precision and efficiency. Point mutations account for 58% of disease-causing genetic mutations in humans, and single nucleotide variants are an important cause of tumorigenesis, and the advent of base editors offers new hope for the study or treatment of such diseases. Although it has some limitations, base editors have been continuously improved in terms of editing efficiency, specificity, and product purity since their development. In this review, we examine the main base editing technologies and discuss their applications and prospects in tumor research and therapy, as well as elaborate on their mode of delivery.

Characteristics, Recombination Methods, and Applications Progresses of Split-Cas9 System.

CRISPR technology has been used to revolutionize various facets of life sciences because of its potent gene editing capabilities. In particular, CRISPR technology is anticipated to be used to cure congenital disorders, and malignant cancers brought on by gene mutation. In this article, we introduce a Split-Cas9 system, in which Cas9 protein is split into two or more parts and recombined in cells to function specific induction circumstances. Split-Cas9 system can improve the therapeutic index of CRISPR technology by splitting Cas9 proteins into small fragments, thus enhancing their compatibility with virus vectors and precise temporal and spatial control. This article examined the combination mode of Split-Cas9 system, contrasted the differences in its split sites and activity efficiency, and discussed the use and clinical transformation in vivo and in vitro.

Exploring disease interrelationships in older inpatients: a single-centre, retrospective study.

Background: Comorbidity is a common phenomenon in the older population; it causes a heavy burden on societies and individuals. However, the relevant evidence, especially in the southwestern region of China, is insufficient. Objectives: We aimed to examine current comorbidity characteristics as well as correlations among diseases in individuals aged >60 years. Design: Retrospective study. Methods: We included records of 2,995 inpatients treated at the Gerontological Department of Sichuan Geriatric Hospital from January 2018 to February 2022. The patients were divided into groups according to sex and age. Diseases were categorised based on the International Classification of Diseases and their Chinese names. We calculated the age-adjusted Charlson Comorbidity Index (ACCI), categorised diseases using the China Health and Retirement Longitudinal Study questionnaire, and visualised comorbidity using web graphs and the Apriori algorithm. Results: The ACCI was generally high, and it increased with age. There were significant differences in the frequency of all diseases across age groups, especially in individuals aged ≥90 years. The most common comorbid diseases were liver diseases, stomach or other digestive diseases, and hypertension. Strong correlations between the most common digestive diseases and hypertension were observed. Conclusion: Our findings provide insights into the current situation regarding comorbidity and the correlations among diseases in the older population. We expect our findings to inform future research directions as well as policies regarding general clinical practice and public health, especially for medical consortiums.

Coaxial radiography guided puncture technique for percutaneous transforaminal endoscopic lumbar discectomy: A randomized control trial.

BACKGROUND: The coaxial radiography-guided puncture technique (CR-PT) is a novel technique for endoscopic lumbar discectomy. As the X-ray beam and the puncturing needle are maintained in a parallel and coaxial direction, the X-ray beam can be used to guide the trajectory angle, facilitating the choice of the puncture site and providing real-time guidance. This puncture technique offers numerous advantages over the conventional anterior-posterior and lateral radiography-guided puncture technique (AP-PT), especially in cases of herniated lumbar discs with a hypertrophied transverse process or articular process, high iliac crest, and narrowed intervertebral foramen. AIM: To confirm whether CR-PT is a superior approach to percutaneous transforaminal endoscopic lumbar discectomy compared to AP-PT. METHODS: In this parallel, controlled, randomized clinical trial, herniated lumbar disc patients appointed to receive percutaneous endoscopic lumbar discectomy treatment were recruited from the Pain Management Department of the Affiliated Hospital of Xuzhou Medical University and Nantong Hospital of Traditional Chinese Medicine. Sixty-five participants were enrolled and divided into either a CR-PT group or an AP-PT group. The CR-PT group underwent CR-PT, and the AP-PT group underwent AP-PT. The number of fluoroscopies during puncturing, puncture duration (min), surgery duration (min), VAS score during puncturing, and puncture success rate were recorded. RESULTS: Sixty-five participants were included, with 31 participants in the CR-PT group and 34 in the AP-PT group. One participant in the AP-PT group dropped out due to unsuccessful puncturing. The number of fluoroscopies [median (P25, P75)] was 12 (11, 14) in the CR-PT group vs 16 (12, 23) in the AP-PT group, while the puncture duration (mean ± SD) was 20.42 ± 5.78 vs 25.06 ± 5.46, respectively. The VAS score was 3 (2, 4) in the CR-PT group vs 3 (3, 4) in the AP-PT group. Further subgroup analysis was performed, considering only the participants with L5/S1 segment herniation: 9 patients underwent CR-PT, and 9 underwent AP-PT. The number of fluoroscopies was 11.56 ± 0.88 vs 25.22 ± 5.33; the puncture duration was 13.89 ± 1.45 vs 28.89 ± 3.76; the surgery duration was 105 (99.5, 120) vs 149 (125, 157.5); and the VAS score was 2.11 ± 0.93 vs 3.89 ± 0.6, respectively. All the above outcomes demonstrated statistical significance (P < 0.05), favoring the CR-PT treatment. CONCLUSION: CR-PT is a novel and effective technique. As opposed to conventional AP-PT, this technique significantly improves puncture accuracy, shortens puncture time and operation time, and reduces pain intensity during puncturing.

Paving a way to treat spastic paraplegia 50.

Spastic paraplegia 50 (SPG50) is a rare neurodegenerative disease caused by loss-of-function mutations in AP4M1. There are no effective treatments for SPG50 or any other type of SPG, and current treatments are limited to symptomatic management. In this issue of the JCI, Chen et al. provide promising data from preclinical studies that evaluated the efficacy and safety profiles of an AAV-mediated AP4M1 gene replacement therapy for SPG50. AAV/AP4M1 gene replacement partly rescued functional defects in SPG50 cellular and mouse models, with acceptable safety profiles in rodents and monkeys. This work represents a substantial advancement in therapeutic development of SPG50 treatments, establishing the criteria for taking AAV9/AP4M1 gene therapy to clinical trials.