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Porcine epidemic diarrhea virus (PEDV) is one of the most devastating diseases affecting the pig industry globally. Due to the emergence of novel strains, no effective vaccines are available for prevention and control. Investigating the pathogenic mechanisms of PEDV may provide insights for creating clinical interventions. This study constructed and expressed eukaryotic expression vectors containing PEDV proteins (except NSP11) with a 3' HA tag in Vero cells. The subcellular localization of PEDV proteins was examined using endogenous protein antibodies to investigate their involvement in the viral life cycle, including endocytosis, intracellular trafficking, genome replication, energy metabolism, budding, and release. We systematically analyzed the potential roles of all PEDV viral proteins in the virus life cycle. We found that the endosome sorting complex required for transport (ESCRT) machinery may be involved in the replication and budding processes of PEDV. Our study provides insight into the molecular mechanisms underlying PEDV infection. IMPORTANCE: The global swine industry has suffered immense losses due to the spread of PEDV. Currently, there are no effective vaccines available for clinical protection. Exploring the pathogenic mechanisms of PEDV may provide valuable insights for clinical interventions. This study investigated the involvement of viral proteins in various stages of the PEDV lifecycle in the state of viral infection and identified several previously unreported interactions between viral and host proteins. These findings contribute to a better understanding of the pathogenic mechanisms underlying PEDV infection and may serve as a basis for further research and development of therapeutic strategies., Competing Interests: Declaration of competing interest The authors declare that have no competing interests., (Copyright © 2024. Published by Elsevier B.V.)

To determine a rapid and accurate method for locating the keypoint and "keyhole" in the suboccipital retrosigmoid keyhole approach. (1) Twelve adult skull specimens were selected to locate the anatomical landmarks on the external surface of the skull.The line between the infraorbital margin and superior margin of the external acoustic meatus was named the baseline. A coordinate system was established using the baseline and its perpendicular line through the top point of diagastric groove.The perpendicular distance (x), and the horizontal distance (y) between the central point of the "keyhole" and the top point of the digastric groove in that coordinate system were measured. The method was applied to fresh cadaveric specimens and 53 clinical cases to evaluate its application value. (1) x and y were 14.20 ± 2.63 mm and 6.54 ± 1.83 mm, respectively (left) and 14.95 ± 2.53 mm and 6.65 ± 1.61 mm, respectively (right). There was no significant difference between the left and right sides of the skull (P > 0.05). (2) The operative area was satisfactorily exposed in the fresh cadaveric specimens, and no venous sinus injury was observed. (3) In clinical practice, drilling did not cause injury to venous sinuses, the mean diameter of the bone windows was 2.0-2.5 cm, the mean craniotomy time was 26.01 ± 3.46 min, and the transverse and sigmoid sinuses of 47 patients were well-exposed. We propose a "one point, two lines, and two distances" for "keyhole" localization theory, that is we use the baseline between the infraorbital margin and superior margin of the external acoustic meatus and the perpendicular line to the baseline through the top point of the digastric groove to establish a coordinate system. And the drilling point was 14.0 mm above and 6.5 mm behind the top point of the digastric groove in the coordinate system., (© 2024. The Author(s).)

Background: Cardioneuroablation (CNA) has shown encouraging results in patients with vasovagal syncope (VVS). However, data on different subtypes was scarce., Methods: This observational study retrospectively enrolled 141 patients [mean age: 40 ± 18 years, 51 males (36.2%)] with the diagnosis of VVS. The characteristics among different types of VVS and the outcomes after CNA were analyzed., Results: After a mean follow-up of 4.3 ± 1.5 years, 41 patients (29.1%) experienced syncope/pre-syncope events after CNA. Syncope/pre-syncope recurrence significantly differed in each subtype ( P = 0.04). The cardioinhibitory type of VVS had the lowest recurrence rate after the procedure ( n = 6, 16.7%), followed by mixed ( n = 26, 30.6%) and vasodepressive ( n = 9, 45.0%). Additionally, a significant difference was observed in the analyses of the Kaplan-Meier survival curve ( P = 0.02). Syncope/pre-syncope burden was significantly reduced after CNA in the vasodepressive type ( P < 0.01). Vasodepressive types with recurrent syncope/pre-syncope after CNA have a lower baseline deceleration capacity (DC) level than those without (7.4 ± 1.0 ms vs. 9.0 ± 1.6 ms, P = 0.01). Patients with DC < 8.4 ms had an 8.1 (HR = 8.1, 95% CI: 2.2-30.0, P = 0.02) times risk of syncope/pre-syncope recurrence after CNA compared to patients with DC ≥ 8.4 ms, and this association still existed after adjusting for age and sex (HR = 8.1, 95% CI: 2.2-30.1, P = 0.02)., Conclusions: Different subtypes exhibit different event-free rates. The vasodepressive type exhibited the lowest event-free rate, but those patients with DC ≥ 8.4 ms might benefit from CNA., (© 2024 JGC All rights reserved; www.jgc301.com.)

Accurate identification of single nucleotide variants (SNVs) in key driver genes holds a significant value for disease diagnosis and treatment. Fluorescent probes exhibit tremendous potential in specific, high-resolution, and rapid detection of SNVs. However, additional steps are required in most post-PCR assays to convert double-stranded DNA (dsDNA) products into single-stranded DNA (ssDNA), enabling them to possess hybridization activity to trigger subsequent reactions. This process not only prolongs the complexity of the experiment but also introduces the risk of losing target information. In this study, we proposed two strategies for enriching active double-stranded DNA, involving PCR based on obstructive groups and cleavable units. Building upon this, we explored the impact of modified units on the strand displacement reaction (SDR) and assessed their discriminatory efficacy for mutations. The results showed that detection of low variant allele frequencies (VAF) as low as 0.1% can be achieved. The proposed strategy allowed orthogonal identification of 45 clinical colorectal cancer tissue samples with 100% specificity, and the results were generally consistent with sequencing results. Compared to existing methods for enriching active targets, our approach offers a more diverse set of enrichment strategies, characterized by the advantage of being simple and fast and preserving original information to the maximum extent. The objective of this study is to offer an effective solution for the swift and facile acquisition of active double-stranded DNA. We anticipate that our work will facilitate the practical applications of SDR based on dsDNA.

Background: The emergence of DNA nanotechnology has enabled the systematic design of diverse bionic dissipative behaviors under the precise control of nucleic acid nanodevices. Nevertheless, when compared to the dissipation observed in robust living systems, it is highly desirable to enhance the anti-interference for artificial DNA dissipation to withstand perturbations and facilitate repairs within the complex biological environments., Results: In this study, we introduce strategically designed "trash cans" to facilitate kinetic control over interferences, transforming the stochastic binding of individual components within a homogeneous solution into a competitive binding process. This approach effectively eliminates incorrect binding and the accumulation of systemic interferences while ensuring a consistent pattern of energy fluctuation from response to silence. Remarkably, even in the presence of numerous interferences differing by only one base, we successfully achieve complete system reset through multiple cycles, effectively restoring the energy level to a minimum., Significance: The system was able to operate stably without any adverse effect under conditions of irregular interference, high-abundance interference, and even multiplex interferences including DNA and RNA crosstalk. This work not only provides an effective paradigm for constructing robust DNA dissipation systems but also greatly broadens the potential of DNA dissipation for applications in high-precision molecular recognition and complex biological reaction networks., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Detecting low-frequency DNA mutations hotspots cluster is critical for cancer diagnosis but remains challenging. Quantitative PCR (qPCR) is constrained by sensitivity, and allele-specific PCR is restricted by throughput. Here we develop a long blocker displacement amplification (LBDA) coupled with qPCR for ultrasensitive and multiplexed variants detection. By designing long blocker oligos to perfectly match wildtype sequences while mispairing with mutants, long blockers enable 14-44 nt enrichment regions which is 2-fold longer than normal BDA in the experiments. For wild template with a specific nucleotide, LBDA can detect different mutation types down to 0.5 % variant allele frequency (VAF) in one reaction, with median enrichment fold of 1,000 on 21 mutant DNA templates compared to the wild type. We applied LBDA-qPCR to detect KRAS and NRAS mutation hotspots, utilizing a single plex assay capable of covering 81 mutations and tested in synthetic templates and colorectal cancer tissue samples. Moreover, the mutation types were verified through Sanger sequencing, demonstrating concordance with results obtained from next generation sequencing. Overall, LBDA-qPCR provides a simple yet ultrasensitive approach for multiplexed detection of low VAF mutations hotspots, presenting a powerful tool for cancer diagnosis and monitoring., (© 2024 Wiley‐VCH GmbH.)

The aim of this study is to propose a new diagnostic model based on "segmentation + classification" to improve the routine screening of Thyroid nodule ultrasonography by utilizing the key domain knowledge of medical diagnostic tasks. A Multi-scale segmentation network based on a pyramidal pooling structure of multi-parallel void spaces is proposed. First, in the segmentation network, the exact information of the underlying feature space is obtained by an Attention Gate. Second, the inflated convolutional part of Atrous Spatial Pyramid Pooling (ASPP) is cascaded for multiple downsampling. Finally, a three-branch classification network combined with expert knowledge is designed, drawing on doctors' clinical diagnosis experience, to extract features from the original image of the nodule, the regional image of the nodule, and the edge image of the nodule, respectively, and to improve the classification accuracy of the model by utilizing the Coordinate attention (CA) mechanism and cross-level feature fusion. The Multi-scale segmentation network achieves 94.27%, 93.90% and 88.85% of mean precision (mPA), Dice value (Dice) and mean joint intersection (MIoU), respectively, and the accuracy, specificity and sensitivity of the classification network reaches 86.07%, 81.34% and 90.19%, respectively. Comparison tests show that this method outperforms the U-Net, AGU-Net and DeepLab V3+ classical models as well as the nnU-Net, Swin UNetr and MedFormer models that have emerged in recent years. This algorithm, as an auxiliary diagnostic tool, can help physicians more accurately assess the benign or malignant nature of Thyroid nodules. It can provide objective quantitative indicators, reduce the bias of subjective judgment, and improve the consistency and accuracy of diagnosis. Codes and models are available at https://github.com/enheliang/Thyroid-Segmentation-Network.git., Competing Interests: Competing interestsThe authors declare no competing interest., (© Korean Society of Medical and Biological Engineering 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Background: Adjuvants may enhance the efficacy of vaccines. however, the efficacy of adjuvant-associated COVID-19 vaccines (ACVs) remains unclear since the emergence of the COVID-19 pandemic. This study aimed to address this gap by conducting a systematic review and meta-analysis of the efficacy of ACVs against Severe Acute Respiratory Syndrome Coronavirus 2 CoV (SARS-CoV-2) variants of concern (VOC)., Methods: A systematic search was conducted of randomized controlled trials (RCTs) evaluating the vaccine efficacy (VE) of ACVs against VOC (alpha, beta, gamma, delta, or Omicron), up to May 27, 2023. The DerSimonian-Laird random-effects model was used to assess VE with 95% confidence intervals (CI) through meta-analysis. Cochrane Risk of Bias tools were used to assess the risk of bias in RCTs., Results: Eight RCTs with 113,202 participants were included in the analysis, which incorporated 4 ACVs [Matrix-M (NVX-CoV2373), Alum (BBV152), CpG-1018/Alum (SCB-2019), and AS03 (CoVLP]). The pooled efficacy of full vaccination with ACVs against VOC was 88.0% (95% CI: 83.0-91.5). Full vaccination was effective against Alpha, Beta, Delta, and Gamma variants, with VE values of 93.66% (95% CI: 86.5-100.74), 64.70% (95% CI: 41.87-87.54), 75.95% (95% CI: 67.9-83.99), and 91.26% (95% CI: 84.35-98.17), respectively. Currently, there is a lack of RCT evidence regarding the efficacy of ACVs against the Omicron variant., Conclusion: In this meta-analysis, it should be that full vaccination with ACVs has high efficacy against Alpha or Gamma variants and moderate efficacy against Beta and Delta variants. Notably, with the exception of the aluminum-adjuvanted vaccine, the other ACVs had moderate to high efficacy against the SARS-CoV-2 variant. This raises concerns about the effectiveness of ACVs booster vaccinations against Omicron., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2024 the Author(s). Published by Wolters Kluwer Health, Inc.)

Nonenzymatic self-assembly circuit utilizing hairpin substrates has been developed to be a powerful tool for information transduction, amplification and computation. However, the sensitivity, stability and application of this circuit are impeded by the presence of leakage which refers to undesired triggering in the absence of input. Herein, we proposed a movable toehold principle to suppress leakage and accelerate the catalytic reaction through removing partial hairpin toehold responsible for the leakage and transferring it to the catalyst. With movable toehold, catalytic hairpin assembly (called mtCHA) exhibited an excellent signal-to-background ratio of over 100, high robustness and improved specificity. In more complex circuit, including proximity recognition, signal amplification of small molecules (such as ATP), logic network, autocatalysis circuit and two-layer cascade circuit, mtCHA also demonstrated satisfactory performance. Our findings suggest that mtCHA holds great potential for broader applications, and the approach of repurposing harmful fragments into beneficial candidates can provide valuable insights for other chemical systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Knee osteoarthritis (KOA) stands as a degenerative ailment with a substantial and escalating prevalence. The practice of traditional Chinese non-pharmacological therapy has become a prevalent complementary and adjunctive approach. A mounting body of evidence suggests its efficacy in addressing KOA. Recent investigations have delved into its underlying mechanism, yielding some headway. Consequently, this comprehensive analysis seeks to encapsulate the clinical application and molecular mechanism of traditional Chinese non-pharmacological therapy in KOA treatment. The review reveals that various therapies, such as acupuncture, electroacupuncture, warm needle acupuncture, tuina, and acupotomy, primarily target localized knee components like cartilage, subchondral bone, and synovium. Moreover, their impact extends to the central nervous system and intestinal flora. More perfect experimental design and more comprehensive research remain a promising avenue in the future., Competing Interests: The authors report no conflicts of interest in this work., (© 2024 Liu et al.)

Multidrug-resistant bacterial infections, especially those caused by multidrug-resistant Escherichia coli ( E. coli ) bacteria, are an ever-growing threat because of the shrinking arsenal of efficacious antibiotics. Therefore, it is urgently needed to develop a kind of novel, long-term antibacterial agent effectively overcome resistant bacteria. Herein, we present a novel designed antibacterial agent-6-Aza-2-thiothymine-capped gold nanoclusters (ATT-AuNCs), which show excellent antibacterial activity against multidrug-resistant E. coli bacteria. The prepared AuNCs could permeabilize into the bacterial cell membrane via binding with a bivalent cation (e.g., Ca 2+ ), followed by the generation of reactive oxygen species (e.g., • OH and • O 2- ), ultimately resulting in protein leakage from compromised cell membranes, inducing DNA damage and upregulating pro-oxidative genes intracellular. The AuNCs also speed up the wound healing process without noticeable hemolytic activity or cytotoxicity to erythrocytes and mammalian tissue. Altogether, the results indicate the great promise of ATT-AuNCs for treating multidrug-resistant E. coli bacterial infection., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

Background: The benefits and risks of adjuvant-associated COVID-19 vaccines (ACVs) are unclear. The study aimed to assess the immunogenicity and safety of ACVs compared with controls (placebo or the same vaccine without adjuvants [NACVs])., Methods: Randomized controlled trials sourced from PubMed, EMBASE, Web of Science, and Cochrane Library were systematically reviewed. Evaluators extracted information independently. The evidence quality was assessed using random-effects models. The risk of bias was assessed using the Cochrane Risk of Bias tool., Results: Of the 33 studies, 27 analyzed immunogenicity (n = 9069, ACVs group; n = 3757, control), and 26 analyzed safety (n = 58669, ACVs groups; n = 30733 control). Compared with controls, full vaccination with ACVs produced significant immune responses (relative risk [RR] of seroneutralization reaction, 12.3; 95 % confidence interval [95 % CI], 6.92-21.89; standardized mean deviation of geometric mean titer 3.96, 95 % CI, 3.35-4.58). Additionally, ACVs produced significant immunoreactivity compared with NACVs only ( P  < 0.05). Furthermore, full vaccination with ACVs significantly increased the risk of local and systemic adverse reactions (AEs) compared with controls. However, vaccination with ACVs did not significantly increase the risk of systemic and localized AEs compared with vaccination with NACVs only (P > 0.05). It was observed that ACVs had a lower risk of all-cause mortality than controls (RR, 0.51; 95 % CI 0.30-0.87). It was further found that ACVs produced nAb response against all sublines of the Omicron variant, but the antibody titers were lower than those for the SARS-CoV-2 original strain., Conclusions: The findings of this meta-analysis demonstrate that ACVs may have a superior effect and an acceptable safety in preventing COVID-19. Although these results suggest the potential of ACVs, further studies are required., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Objectives: Most Asian countries have employed Chinese medicine (CM) and Western medicine to treat lumbar spinal stenosis (LSS). Evidence synthesis and comparison of effectiveness are difficult since outcomes examined and presented through trials possess heterogeneity. This study aimed to solve the outcome problems for CM clinical trials in LSS by building a core outcome set (COS)., Methods: To achieve an agreement on a set of core outcome domains, a four-phase study was carried out. First, we identified candidate outcome domains by systematically reviewing trials. In addition, we identified outcome domains associated with patients by conducting semistructured interviews with patients. Next, outcome domains were processed through a national two-round Delphi survey, in which 18 patients and 21 experts were recruited. Finally, the above domains were converted as a core outcome domain set based on a consensus meeting, in which 24 stakeholders were recruited., Results: Seventeen outcome subdomains were identified by the systematic review and interviews. The Delphi survey assigned a priority to four outcome domains in the first round and four outcomes additionally in the second round. The core outcome domains were determined through discussion and redefinition of outcomes in the consensus meeting: pain and discomfort, health-related quality of life, lumbar function, activities of daily living, measures of walking, patient global assessment, adverse events and CM-specific outcomes., Conclusion: COS-CM-LSS is likely to enhance the consistency of outcomes reported in clinical trials. In-depth research should be conducted for the exploration of the best methods to examine the above outcomes., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: The mortality rate from septic shock in patients with hematological malignancies (HMs) remains significantly higher than that in patients without HMs. A longer resuscitation time would definitely be harmful because of the irreversibly immunocompromised status of the patients. Shortening the resuscitation time through continuous renal replacement therapy (CRRT) with oXiris ® would be an attractive strategy in managing such patients., Aim: To explore the effects of CRRT and oXiris ® in shortening the resuscitation time and modifying the host response by reducing inflammation mediator levels., Methods: Forty-five patients with HM were diagnosed with septic shock and underwent CRRT between 2018 and 2022. Patients were divided into two groups based on the hemofilter used for CRRT (oXiris ® group, n = 26; M150 group, n = 19). We compared the number of days of negative and total fluid balance after 7 d of CRRT between the groups. The heart rate, norepinephrine dose, Sequential Organ Failure Assessment (SOFA) score, and blood lactic acid levels at different time points in the two groups were also compared. Blood levels of inflammatory mediators in the 26 patients in the oXiris ® group were measured to further infer the possible mechanism., Results: The average total fluid balance after 7 d of CRRT in the oXiris ® group was significantly lower than that of patients in the M150 hemofilter group. The SOFA scores of patients after CRRT with oXiris ® therapy were significantly lower than those before treatment on day 1 (d1), d3 and d7 after CRRT; these parameters were also significantly lower than those of the control group on d7. The lac level after oXiris ® therapy was significantly lower than that before treatment on d3 and d7 after CRRT. There were no significant differences in the above parameters between the two groups at the other time points. In the oXiris ® group, procalcitonin levels decreased on d7, whereas interleukin-6 and tumor necrosis factor levels decreased significantly on d3 and d7 after treatment., Conclusion: CRRT with oXiris ® hemofilter may improve hemodynamics by reducing inflammatory mediators and playing a role in shortening the resuscitation period and decreasing total fluid balance in the resuscitation phases., Competing Interests: Conflict-of-interest statement: The authors declare no conflicts of interest., (©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.)

Porcine enteric alphacoronavirus (PEAV) is a new bat HKU2-like porcine coronavirus, and its endemic outbreak has caused severe economic losses to the pig industry. Its broad cellular tropism suggests a potential risk of cross-species transmission. A limited understanding of PEAV entry mechanisms may hinder a rapid response to potential outbreaks. This study analyzed PEAV entry events using chemical inhibitors, RNA interference, and dominant-negative mutants. PEAV entry into Vero cells depended on three endocytic pathways: caveolae, clathrin, and macropinocytosis. Endocytosis requires dynamin, cholesterol, and a low pH. Rab5, Rab7, and Rab9 GTPases (but not Rab11) regulate PEAV endocytosis. PEAV particles colocalize with EEA1, Rab5, Rab7, Rab9, and Lamp-1, suggesting that PEAV translocates into early endosomes after internalization, and Rab5, Rab7, and Rab9 regulate trafficking to lysosomes before viral genome release. PEAV enters porcine intestinal cells (IPI-2I) through the same endocytic pathway, suggesting that PEAV may enter various cells through multiple endocytic pathways. This study provides new insights into the PEAV life cycle. IMPORTANCE Emerging and reemerging coronaviruses cause severe human and animal epidemics worldwide. PEAV is the first bat-like coronavirus to cause infection in domestic animals. However, the PEAV entry mechanism into host cells remains unknown. This study demonstrates that PEAV enters into Vero or IPI-2I cells through caveola/clathrin-mediated endocytosis and macropinocytosis, which does not require a specific receptor. Subsequently, Rab5, Rab7, and Rab9 regulate PEAV trafficking from early endosomes to lysosomes, which is pH dependent. The results advance our understanding of the disease and help to develop potential new drug targets against PEAV.

In this work, we propose a hairpin probe-mediated exponential amplification reaction (HEAR) strategy that combines DNA strand displacement with a "who triggers, who gets generated" mode, providing excellent single-base discrimination and a reduced background signal. The detection limit is 19 aM, which is reduced by 3 orders of magnitude compared to traditional exponential amplification approaches. This one-pot strategy also exhibits a wide dynamic range, high specificity and short detection time. It is expected to become a powerful tool for clinical diagnosis.

This study explored the role of P2X7 receptors in spinal cord astrocytes in the electroacupuncture-induced inhibition of visceral hypersensitivity (VH) in rats with irritable bowel syndrome (IBS). Visceral hypersensitivity of IBS was intracolonically induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS). Visceromotor responses to colorectal distension (CRD-20,40,60,80 mmHg) and abdominal withdrawal reflex scoring (AWRs) were recorded after electroacupuncture at bilateral Zusanli (ST36) and Sanyinjiao (SP6) acupoints to evaluate the analgesic effect of electroacupuncture on visceral pain in rats with IBS. Fluorocitric acid (FCA), an astrocyte activity inhibitor, was injected intrathecally before electroacupuncture intervention and AWRs were recorded. Western blot and real-time qPCR were used to detect the expression of NMDA and P2X7 receptor to observe the regulation effect of electroacupuncture on NMDA receptor in the spinal cord of rats with visceral hypersensitivity. Intrathecal injection of P2X7 agonist or antagonist was administered before electroacupuncture treatment. To observe the effect of P2X7 receptor in spinal astrocytes on the inhibition of visceral hyperalgesia by electroacupuncture, the changes of AWR score, NMDA receptor in the spinal cord, and GFAP expression in astrocytes were detected. Inflammation of the colon had basically subsided at day 21 post-TNBS; persistent visceral hypersensitivity could be suppressed by electroacupuncture. This analgesic effect could be inhibited by FCA. The analgesic effect, downregulation of NMDA receptor NR1 subunit, and P2X7 protein of electroacupuncture were all reversed by FCA. P2X7 receptor antagonist A740003 can cooperate with EA to carry out analgesic effect in rats with visceral pain and downregulate the expression of NR1, NR2B, and GFAP in spinal dorsal horn. However, the P2X7 receptor agonist BzATP could partially reverse the analgesic effect of EA, inhibiting the downregulatory effect of EA on the expression of NR1, NR2B, and GFAP. These results indicate that EA may downregulate the expression of the NMDA receptor by inhibiting the P2X7 receptor in the spinal cord, thereby inhibiting spinal cord sensitization in IBS rats with visceral pain, in which astrocytes are an important medium., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

The incidence of thyroid nodules is increasing year by year. Accurate determination of benign and malignant nodules is an important basis for formulating treatment plans. Ultrasonography is the most widely used methodology in the diagnosis of benign and malignant nodules, but diagnosis by doctors is highly subjective, and the rates of missed diagnosis and misdiagnosis are high. To improve the accuracy of clinical diagnosis, this paper proposes a new diagnostic model based on deep learning. The diagnostic model adopts the diagnostic strategy of localization-classification. First, the distribution laws of the nodule size and nodule aspect ratio are obtained through data statistics, a multiscale localization network structure is a priori designed, and the nodule aspect ratio is obtained from the positioning results. Then, uncropped ultrasound images and nodule area image are correspondingly input into a two-way classification network, and an improved attention mechanism is used to enhance the feature extraction performance. Finally, the deep features, the shallow features, and the nodule aspect ratio are fused, and a fully connected layer is used to complete the classification of benign and malignant nodules. The experimental dataset consists of 4021 ultrasound images, where each image has been labeled under the guidance of doctors, and the ratio of the training set, validation set, and test set sizes is close to 3:1:1. The experimental results show that the accuracy of the multiscale localization network reaches 93.74%, and that the accuracy, specificity, and sensitivity of the classification network reach 86.34%, 81.29%, and 90.48%, respectively. Compared with the champion model of the TNSCUI 2020 classification competition, the accuracy rate is 1.52 points higher. Therefore, the network model proposed in this paper can effectively diagnose benign and malignant thyroid nodules., (© 2023. The Author(s).)

CRISPR (clustered regularly interspaced short palindromic repeats) technology has achieved great breakthroughs in terms of convenience and sensitivity; it is becoming the most promising molecular tool. However, only two CRISPR activation modes (single and double stranded) are available, and they have specificity and universality bottlenecks that limit the application of CRISPR technology in high-precision molecular recognition. Herein, we proposed a novel CRISPR/Cas12a unrestricted activation mode to greatly improve its performance. The new mode totally eliminates the need for a protospacer adjacent motif and accurately activates Cas12a through toehold-mediated strand displacement and branch migration, which is highly universal and ultra-specific. With this mode, we discriminated all mismatch types and detected the EGFR T790M and L858R mutations in very low abundance. Taken together, our activation mode is deeply incorporated with DNA nanotechnology and extensively broadens the application boundaries of CRISPR technology in biomedical and molecular reaction networks., (© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Digestive system cancers account for nearly half of all cancers around the world and have a high mortality rate. Cell culture and animal models represent cornerstones of digestive cancer research. However, their ability to enable cancer precision medicine is limited. Cell culture models cannot retain the genetic and phenotypic heterogeneity of tumors and lack tumor microenvironment (TME). Patient-derived xenograft mouse models are not suitable for immune-oncology research. While humanized mouse models are time- and cost-consuming. Suitable preclinical models, which can facilitate the understanding of mechanisms of tumor progression and develop new therapeutic strategies, are in high demand. This review article summarizes the recent progress on the establishment of TME by using tumor organoid models and microfluidic systems. The main challenges regarding the translation of organoid models from bench to bedside are discussed. The integration of organoids and a microfluidic platform is the emerging trend in drug screening and precision medicine. A future prospective on this field is also provided., Competing Interests: Declaration of Competing Interest The authors declare that there are no conflicts of interest., (Copyright © 2022. Published by Elsevier Inc.)

Objective: To explore whether acupuncture combined with moxibustion could inhibit epithelialmesenchymal transition in Crohn's disease by affecting the transforming growth factor β 1 (TGF- β 1)/Smad3/Snail pathway., Methods: Sixty-three patients with Crohn's disease were randomly divided into an observation group (31 cases) receiving moxibustion at 43 °C combined with acupuncture, and a control group (32 cases) receiving moxibustion at 37 °C combined with sham acupuncture using a random number table. Patients were treated for 12 weeks. Crohn's Disease Activity Index (CDAI) was used to evaluate disease activity. Hematoxylin-eosin staining and transmission electron microscopy were utilized to observe the morphological and ultrastructural changes. Immunohistochemistry was used to detect the expression of transforming growth factor β 1 (TGF-β 1), T β R1, T β R2, Smad3, Snail, E-cadherin and fibronectin in intestinal mucosal tissues., Results: The decrease of the CDAI score, morphological and ultrastructural changes were more significant in observation group. The expression levels of TGF- β 1, Tβ R2, Smad3, and Snail in the observation group were significantly lower than those before the treatment (P<0.05 or P<0.01). After treatment, the expression levels of TGF-β 1, TβR2, and Snail in the observation group were significantly lower than those in the control group (all P<0.05); compared with the control group, the expression of fibronectin in the observation group was significantly decreased, and the expression of E-cadherin was significantly increased (all P<0.05)., Conclusions: Moxibustion at 43 °C combined with acupuncture may suppress TGF-β 1/Smad3/Snail pathway-mediated epithelial-mesenchymal transition of intestinal epithelial cells in Crohn's disease patients by inhibiting the expression levels of TGF-β 1, Tβ R2, Smad3, and Snail. (Registration No. ChiCTR-IIR-16007751)., (© 2022. The Chinese Journal of Integrated Traditional and Western Medicine Press and Springer-Verlag GmbH Germany, part of Springer Nature.)

Currently, developing an effective and easy-to-operate signal amplification assay to detect the trace-amount miRNAs in serum remains a significant challenge. Herein, an ultrasensitive CeO 2 @Ag hybrid nanoflower (CeO 2 @Ag HNF)-labeled electrochemical biosensor was developed for sensing miRNA, based on a target-feedback rolling-cleavage (TFRC) signal amplifier. CeO 2 @Ag HNFs possessing a unique three-dimensional layered structure were synthesized without any complex reaction conditions, such as heating and vacuum. The bimetallic nanoflowers provided a large surface area and excellent CAT-like activity, thereby enhancing electrochemical performance. Based on the principle of branched catalytic hairpin assembly, target miRNA could continuously trigger the assembly of branched junctions with ends composed of DNAzyme. The activated DNAzyme was able to cleave the hairpin substrate efficiently and release to capture more CeO 2 @Ag HNFs label. The process combining target-driven branched catalytic hairpin assembly and DNAzyme-assisted rolling cleavage were defined as TFRC signal amplification. This sensitive electrochemical biosensor exhibited good linear relationship of 1 fM - 1 nM with a detection limit down to 0.89 fM. The proposed method is expected to have a promising application in the miRNA-associated fundamental research and clinical diagnosis., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Automatic estimation of the poses of dairy cows over a long period can provide relevant information regarding their status and well-being in precision farming. Due to appearance similarity, cow pose estimation is challenging. To monitor the health of dairy cows in actual farm environments, a multicow pose estimation algorithm was proposed in this study. First, a monitoring system was established at a dairy cow breeding site, and 175 surveillance videos of 10 different cows were used as raw data to construct object detection and pose estimation data sets. To achieve the detection of multiple cows, the You Only Look Once (YOLO)v4 model based on CSPDarkNet53 was built and fine-tuned to output the bounding box for further pose estimation. On the test set of 400 images including single and multiple cows throughout the whole day, the average precision (AP) reached 94.58%. Second, the keypoint heatmaps and part affinity field (PAF) were extracted to match the keypoints of the same cow based on the real-time multiperson 2D pose detection model. To verify the performance of the algorithm, 200 single-object images and 200 dual-object images with occlusions were tested under different light conditions. The test results showed that the AP of leg keypoints was the highest, reaching 91.6%, regardless of day or night and single cows or double cows. This was followed by the AP values of the back, neck and head, sequentially. The AP of single cow pose estimation was 85% during the day and 78.1% at night, compared to double cows with occlusion, for which the values were 74.3% and 71.6%, respectively. The keypoint detection rate decreased when the occlusion was severe. However, in actual cow breeding sites, cows are seldom strongly occluded. Finally, a pose classification network was built to estimate the three typical poses (standing, walking and lying) of cows based on the extracted cow skeleton in the bounding box, achieving precision of 91.67%, 92.97% and 99.23%, respectively. The results showed that the algorithm proposed in this study exhibited a relatively high detection rate. Therefore, the proposed method can provide a theoretical reference for animal pose estimation in large-scale precision livestock farming., Competing Interests: The authors have declared that no competing interests exist.

Objective: We retrospectively assessed the surgical results of PBC with preoperative multimodal image fusion and intraoperative Dyna Computed Tomography (CT) in 24 patients with primary trigeminal neuralgia (PTN) to explore a valuable aid for Percutaneous balloon compression (PBC)., Methods: We studied the data of 24 patients with PTN. All patients underwent PBC and were assessed with preoperative multimodal image fusion [computed tomography (CT) and magnetic resonance imaging (MRI)] and intraoperative Dyna CT in the Department of Neurosurgery of Zhuhai People's Hospital between October 2020 and September 2021. Multimodal image fusion-three-dimensional (3D) reconstruction of CT and MRI data-was performed using 3D-Slicer software, and preoperative evaluation was performed according to the results of image fusion. Dyna CT was used to dynamically observe the position and shape of the metallic hollow introducer and Fogarty catheter and balloon during the operation to guide the operation in real time. We performed follow-up assessments each month and summarized the clinical characteristics, surgical effects, and complications in all patients., Results: Surgery was successful for all patients; the patients reported immediate pain relief. Surgical complications included facial numbness in 24 patients (100%), mild masseter weakness in three (12.5%), herpes zoster in three (12.5%), and balloon rupture in one (4.2%). None of the patients had serious surgical complications. The mean follow-up time was 9.6 ± 2.7 months. During the follow-up period, 22 patients (91.7%) experienced no recurrence of pain, and two patients (8.3%) experienced recurrence of pain, of which one underwent secondary PBC surgery., Conclusions: Preoperative multimodal image reconstruction can help fully evaluate PBC surgery, clarify the etiology, and predict the volume of contrast medium required during the operation. It provided important assistance for PBC treatment of trigeminal neuralgia patients when preoperative multimodal image fusion is combined with intraoperative Dyna CT., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Liao, Li, Wu, Jian, Yi, Weng and Chen.)

Objective: We aimed to explore a method of precise localization within craniotomy based on skull anatomical landmarks via the suboccipital retrosigmoid approach., Method: Craniometric measurements were taken from 15 adult dry skulls and eight cadaver head specimens. In the anatomical study, the keypoint corresponded to the transverse-sigmoid sinus junction's corresponding point on the external surface of the temporal mastoid process, eight cadaveric heads underwent a simulated craniotomy using the suboccipital retrosigmoid approach. The center of the burr hole is precisely oriented 12 mm vertically above the top point of the mastoid groove based on the line between the infraorbital margin and the upper edge of the external auditory canal. Clinical application was verified in clinical surgery by evaluating the accuracy, safety, rapidity, and minimal invasiveness of the procedure in 29 patients., Result: No venous sinus injuries were observed. Within clinical application, 29 patients underwent craniotomy using the suboccipital retrosigmoid approach. The operative area was clearly exposed in all patients and the microsurgical anatomy of the intracranial region after the dura mater incision was satisfactory. No venous sinus ruptures were observed. The average craniectomy time was 27.02 ± 0.86 min. The diameter of the bone window was 1.7-2.9 cm., Conclusion: We conclude that the method can ensure safe, accurate, and rapid craniotomy with good vision while avoiding injury to the venous sinus., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Jian, Sheng, Li, Li, Weng and Chen.)

Nucleic acid hybridization plays a critical role in medical diagnostics and nanotechnology, but its selectivity and robustness remain to be improved. Here, focusing on double-stranded nucleic acid-based hybridization, we present a series of related strategies. Above all, two simple strategies for enriching toehold-included double-stranded nucleic acids have been proposed. On this basis, two universal hybridization methods with higher selectivity than typical toehold exchange reaction and a long target detection method using short probes to extend the detectable length range are realized. We also provide a double-stranded nucleic acids-catalyzed cycle amplification reaction to improve sensitivity, which has superior interference resistance and excellent discrimination for single-base mismatches. Besides, double-stranded nucleic acids with forked toeholds are used as essential elements to construct a series of logic gates that can evaluate different input combinations. Given the unique advantages of double-stranded nucleic acids, we expect the current work to advance the application of double-stranded nucleic acid-based hybridization in medical diagnostics and nanotechnology., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

DNA strand displacement plays an essential role in the field of dynamic DNA nanotechnology. However, flexible regulation of strand displacement remains a significant challenge. Most previous regulatory tools focused on controllable activation of toehold and thus limited the design flexibility. Here, we introduce a regulatory tool termed cooperative branch migration (CBM), through which DNA strand displacement can be controlled by regulating the complementarity of branch migration domains. CBM shows perfect compatibility with the majority of existing regulatory tools, and when combined with forked toehold, it permits continuous fine-tuning of the strand displacement rate spanning 5 orders of magnitude. CBM manifests multifunctional regulation ability, including rate fine-tuning, continuous dynamic regulation, reaction resetting, and selective activation. To exemplify the powerful function, we also constructed a nested if-function signal processing system on the basis of cascading CBM reactions. We believe that the proposed regulatory strategy would effectively enrich the DNA strand displacement toolbox and ultimately promote the construction of DNA machines of higher complexity in nucleic acid research and biomedical applications.

Seven benzophenone compounds were synthesized in one or two steps, then their antitumor activity was evaluated. The total yields ranged from 9% to 44%. Compounds 3c-5c exhibited obvious antitumor activity. Among them, compounds 3c and 4c exhibited excellent and broad-spectrum antitumor activity. Compound 3c exhibited much stronger inhibitory activities against fourteen cancer cells than cisplatin. In particular, compound 3c exhibited stronger cytotoxicity against hepatocarcinoma SMMC-7721 cells than Taxol, with a half maximal inhibitory concentration (IC 50 ) of approximately 0.111 μM. These results demonstrated that compounds 3c , 4c and 5c were very promising antitumor leads for further structural modification.

N 6 -methyladenosine (m6A) is the most abundant post-transcriptional modification in RNA and has important implications in physiological processes and tumor development. However, sensitive and specific quantification of locus-specific m6A modification levels remains a challenging task. In the present work, a novel m6A-sensitive DNAzyme was utilized to directly detect m6A by coupling with a three-way junction-mediated isothermal exponential CRISPR amplification reaction for the first time. This method was built on the fact that the binding arm of the DNAzyme bound to the specific site and its core structure catalyzed the selective cleavage of unmodified adenine instead of methylated adenines. Subsequently, the intact RNA was identified by the proximity effect of the three-way junction. Enormous amounts of single-stranded DNA products were generated through a combination of SDA and EXPAR for signal amplification. The specific real-time curve of products was recorded through detecting the fluorescence intensity triggered by CRISPR Cas12a. As a result, methylation target of abundance down to 1% was successfully identified. In addition, this strategy could be used for the analysis of cell RNA extracts. Combined with an electrochemical sensor for quantitative detection of RNA methylation, we demonstrated the generality of as-proposed strategy. We envision the present method would provide a new platform for the analysis of m6A in RNA and promote its application in clinical diseases., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Understanding the mechanism of the negative differential resistance (NDR) in transition metal dichalcogenides is essential for fundamental science and the development of electronic devices. Here, the NDR of the current-voltage characteristics was observed based on the glutamine-functionalized WS 2 quantum dots (QDs). The NDR effect can be adjusted by varying the applied voltage range, air pressure, surrounding gases, and relative humidity. A peak-to-valley current ratio as high as 6.3 has been achieved at room temperature. Carrier trapping induced by water molecules was suggested to be responsible for the mechanism of the NDR in the glutamine-functionalized WS 2 QDs. Investigating the NDR of WS 2 QDs may promote the development of memory applications and emerging devices., (© 2021 IOP Publishing Ltd.)

Objective: To explore the precise location of the keypoint during craniotomy using the retrosigmoid keyhole approach. Methods: This study included 20 dry skulls and 10 wet cadaveric specimens. On the inner surface of dry skulls, the junction between the inferior margin of the transverse sinus (ITS) and the posterior margin of the sigmoid sinus (TSJ) was marked. The keypoint (D) was identified as the TSJ's corresponding point on the external surface of the temporal mastoid process (MP). The distance from the keypoint to the top point of the digastric groove, mastoidale, and asterion were noted (AD, BD, CD, respectively). A method to accurately locate the keypoint was developed based on these relationships. The developed method was used on the wet cadaveric specimens to evaluate its accuracy, safety, rapidity, and minimal invasion. Results: No significant difference was found between the AD, BD, and CD of the left and right sides. The drilling point was oriented on a straight line 12 mm above the top point of digastric groove, perpendicular to the Frankfort horizontal plane (FHP). In the cadaveric specimens, the operative area was clearly exposed. No venous sinus rupture occurred. The average craniotomy time was 28.74 ± 3.89 min. Conclusions: A potentially safe, accurate, and rapid craniotomy procedure was developed with the added advantage of preserving the visibility of the operating field and preventing venous sinus injury., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Jian, Sheng, Li, An, Weng and Chen.)

Objective: To investigate the clinical effects of the L-carnitine-astaxanthin compound nutrients Menglankang (MLK) on idiopathic oligospermia (OS) and asthenospermia (AS)., Methods: This study included 73 cases of OS and 220 cases of AS treated with MLK once a bag, bid, for 3 successive months. Before and at 1, 2 and 3 months after treatment, we obtained and analyzed the semen parameters and sperm DNA fragmentation index (DFI) of the patients., Results: Compared with the baseline, the OS patients showed remarkable increases after 1 and 2 months of treatment in the semen volume (［3.07 ± 1.47］ vs ［3.26 ± 1.26］ and ［3.30 ± 1.28］ ml), sperm concentration (［10.96 ± 6.09］ vs ［16.74 ± 11.15］ and ［17.56 ± 9.92］ ×10⁶/ml, P < 0.05), total sperm count (［29.78 ± 17.48］ vs ［52.98 ± 32.07］ and ［57.67 ± 36.98］ ×106, P < 0.05) and the percentages of progressively motile sperm (PMS) (［39.8 ± 11.66］% vs ［45.3 ± 14.03］% and ［46.42 ± 10.69］%, P < 0.05) and morphologically normal sperm (MNS) (［1.71 ± 1.07］% vs ［1.79 ± 0.91］% and ［1.84 ± 0.96］%), and so did the AS patients in PMS (［19.23 ± 8.32］% vs ［25.46 ± 13.86］% and ［27.33 ± 12.88］%, P < 0.05). After 3 months of medication, the OS patients exhibited even more significant increases in the semen volume (［3.63 ± 1.39］ ml) (P < 0.05), sperm concentration (［20.56 ± 14.7］ ×10⁶/ml) (P < 0.05), total sperm count (［66.35 ± 55.91］ ×106) (P < 0.05), PMS (［49.24 ± 13.45］%) (P < 0.05) and MNS (［2.59 ± 0.93］%) (P < 0.05), and so did the AS patients in the semen volume (［3.27 ± 1.42］ vs ［3.85 ± 1.59］ ml, P < 0.05), PMS (［29.11 ± 13.58］%) (P < 0.05) and NMS (［2.01 ± 1.14］% vs ［2.57 ± 1.15］%, P < 0.05). In comparison with the baseline, the sperm DFI was not significantly improved at 1 month after treatment, but remarkably decreased at 2 and 3 months in the OS patients (［25.87 ± 13.76］% vs ［18.66 ± 10.83］% and ［16.48 ± 11.46］%, P < 0.05) and the AS patients as well (［26.40 ± 12.28］% vs ［19.35 ± 11.54］% and ［15.32 ± 10.89］%, P < 0.05)., Conclusions: The L-carnitine-astaxanthin compound nutrients Menglankang can significantly improve the semen quality of the patients with idiopathic oligospermia or asthenospermia.

Sixteen substituted 1-hydroxy-3-methylxanthones were synthesized in one step. The yields ranged from 33 to 76%. Then, the antitumor, antioxidant, anti-tyrosinase, anti-pancreatic lipase, and antifungal activities of compounds 1 - 16 were evaluated. Compounds 10 - 12 and 14 inhibited tyrosinase and pancreatic lipase activity to a certain extent, respectively. Compound 16 exhibited obvious cytotoxicity against fifteen cancer cells, moderate antioxidant activity, and moderate inhibitory activity against Candida albicans . In particular, compound 16 exhibited strong inhibitory activity against A-549 and A549/Taxol cells. These results demonstrated that compounds 10 - 12 , 14 , and 16 are promising leads for further structural modification.[Formula: see text].

In this study, an ultrasensitive electrochemical miRNA biosensor based on toehold mediated strand displacement reaction circuits (SDRCs) and molecular beacon mediated isothermal circular strand displacement polymerization reaction (ICSDPR) has been proposed. During the SDRCs module, the cascade strand displacement reaction induces the recycling of the target let 7a and generation of a large amount of strand A (SA). The SA recognition opens the hairpin capture probe immobilized on the gold electrode, thus, varying the distance between the redox molecules and electrode surface. The primer mediated ICSDPR is observed to further generate a large amount of SA, thus, leading to a reduction in the signal. Considering these merits, the proposed method is observed to exhibit a log-linear linearity from 10 aM to 100 pM and ultrahigh sensitivity towards let 7a down to 6.2 aM, with a capability of distinguishing the let 7a family members, thereby, providing a new electrochemical route for early cancer screening., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Sensitive and specific miRNA detection is essential for the early cancer diagnosis. In this work, we design a fluorescent microRNA biosensor based on exponential amplification reaction (EXPAR) and nicking endonuclease-powered three-dimensional (3-D) bipedal DNA walkers (BDW). Target microRNA initiates EXPAR with the help of polymerase and nicking endonuclease to generate the large number of BDW in solution. The newly generated BDW can be continuously assembled onto polystyrene microsphere track co-modified with fluorescence-labeled DNA strand. Thus, in the presence of nicking endonuclease, the walking machine is activated to produce enhanced fluorescent signal in the supernatant. Besides, we prove that BDW holds the faster walking speed than single-legged DNA walker (SDW) based on comparative study. Under optimal conditions, the proposed amplification method owns a wide linear range from 10 fM to 5 nM with a detection limit down to 5.2 fM. The reaction time of the assay takes about 70 min. The combination of enzyme-assisted EXPAR in solution and enzyme-powered BDW on particle significantly increases the signal amplification efficiency and improves the detection sensitivity. Therefore, our method has enormous potential for the application of BDW-related biosensors., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objective: To observe the effect of acupuncture-moxibustion on negative emotions and plasma tryptophan (Trip)-kynurenine (Kyn) metabolism in the patients with Crohn's disease (CD) at the mild and moderate active stage., Methods: A total of 66 CD patients were randomized into an observation group (33 cases, 1 case dropped off) and a control group (33 cases, 2 cases dropped off). In the observation group, acupuncture was applied in combination with moxibustion. In the control group, the sham-acupuncture was used in combination with sham-moxibustion. In both of the observation group and the control group, acupuncture was applied to Zhongwan (CV 12), Shangjuxu (ST 37), Sanyinjiao (SP 6), Gongsun (SP 4), Hegu (LI 4), Quchi (LI 11), Taixi (KI 3) and Taichong (LR 3), and moxibustion was applied to Tianshu (ST 25) and Zusanli (ST 36). The treatment was given once every two days, 3 times a week, totally for 12 weeks. Separately, before and after treatment, the score of the hospital anxiety-depression scale (HADS) and the score of intestinal core symptoms (degree of abdominal pain and frequency of diarrhea) were observed in the patients of the two groups. The concentration of plasma indoleamine 2,3-dioxygenase 1 (IDO1) and the ratios of Kyn/Trp, QuinA/Kyn, KynA/Kyn and KynA/QuinA were compared between the two groups., Results: Compared with before treatment, the scores of HADS-A and HADS-D in the observation group and the score of HADS-A in the control group were all reduced after treatment ( P <0.01, P <0.05). The scores of abdominal pain degree in the two groups and score of diarrhea frequency in the observation group were all reduced after treatment ( P <0.001). After treatment, the reducing ranges of the score of HADS-A and the scores of abdominal pain degree and diarrhea frequency in the observation group were all larger than the control group ( P <0.01, P <0.05). Compared with before treatment, the plasma IDO1 concentration in the two groups and the ratios of plasma Kyn/Trp and QuinA/Kyn in the observation group were all reduced after treatment ( P <0.001, P <0.05, P <0.01), the ratios of plasma KynA/Kyn and KynA/QuinA were increased after treatment in the observation group ( P <0.05, P <0.01). After treatment, the changes in IDO1 concentration and the ratios of plasma QuinA/Kyn and KynA/QuinA in the observation were larger than the control group ( P ≤0.01, P <0.05). In the observation group, the difference in the ratio of plasma KynA/Kyn before and after treatment was negatively related to the improvement value of HADS-D ( r =-0.67, P <0.05). After treatment, plasma IDO1 concentration was positively related to HADS-A in the observation group ( r =0.65, P <0.05)., Conclusion: Acupuncture and moxibustion relieve the negative emotions of anxiety and depression in CD patients at mild and moderate active stage, which is probably related to the regulation of plasma Trp-Kyn metabolic pathway.

Multiplex PCR encounters difficulties in primer designing with all the primer pairs working at the same annealing temperature. In this study, we have developed a double-strand primer-mediated multiple strand displacement reaction for the detection of SARS-COV-2 ORF, N and E genes (as examples). The double primer is composed of a 5'-modified fluorophore strand, which does not impact polymerase extension and a 3'-modified quencher strand, which cannot impact elongation. At the annealing temperature, the fluorophore strand combined with the template, extended and resulted in fluorescence signal release. Results showed that the double-strand primer relatively exhibits a wide annealing temperature range and good compatibility between three pairs of primers and probes. These merits allow the simple and multiplex real-time fluorescence quantification of nucleic acids. The detection limit was 400 copies/mL, and the detection time was approximately 2 h. In addition to its extreme specificity and simplicity, this method has a wide range of applications such as multiple PCR and SNP detection.

Objective: To investigate the clinical value of the prostate small extracorporeal protein (PSEP) level in the urine in evaluating the therapeutic effect on chronic prostatitis (CP)., Methods: Totally 188 CP patients were treated with minocycline and Ningmitai Capsules in our hospital and regularly returned for follow-up examination from November 2017 to November 2018. Based on the results of treatment after 4 and 8 weeks of medication, we divided the patients into a cured, an effective and an ineffective group and compared the contents of PSEP in the urine samples of the three groups of patients before and after treatment., Results: Compared with the baseline, the PSEP content in the urine after 4 weeks of medication was decreased in the cured group (n = 20) (［3.63 ± 3.81］ vs ［1.16 ± 0.41］ ng/ml, P < 0.05), effective group (n = 85) (［4.13 ± 4.05］ vs ［2.97 ± 2.89］ ng/ml, P > 0.05) and ineffective group (n = 83) (［4.72 ± 2.98］ vs ［3.74 ± 1.31］ ng/ml, P > 0.05), and so was that after 8 weeks of treatment in the cured group (n = 48) (［3.72 ± 3.51］ vs ［0.89 ± 0.37］ ng/ml, P < 0.05), effective group (n = 106) (［4.37 ± 3.93］ vs ［1.83 ± 0.71］ ng/ml, P < 0.05) and ineffective group (n = 34) (［4.61 ± 3.59］ vs ［3.58 ± 1.15］ ng/ml, P > 0.05)., Conclusions: The PSEP level in the urine can be used as an index for clinical evaluation of the therapeutic effect on chronic prostatitis.

New agents with particular specificity toward targeted bacteria and superefficacy in antibacterial activity are urgently needed in facing the crisis of worldwide antibiotic resistance. Herein, a novel strategy by equipping bacteriophage (PAP) with photodynamic inactivation (PDI)-active AIEgens (luminogens with aggregation-induced emission property) was presented to generate a type of AIE-PAP bioconjugate with superior capability for both targeted imaging and synergistic killing of certain species of bacteria. The targeting ability inherited from the bacteriophage enabled the bioconjugates to specifically recognize the host bacteria with preserved infection activity of phage itself. Meanwhile, the AIE characteristic empowered them a monitoring functionality, and the real-time tracking of their interactions with targets was therefore realized via convenient fluorescence imaging. More importantly, the PDI-active AIEgens could serve as powerful in situ photosensitizers producing high-efficiency reactive oxygen species (ROS) under white light irradiation. As a result, selective targeting and synergistic killing of both antibiotic-sensitive and multi-drug-resistant (MDR) bacteria were successfully achieved in in vitro and in vivo antibacterial tests with excellent biocompatibility. This novel AIE-phage integrated strategy would diversify the existing pool of antibacterial agents and inspire the development of promising drug candidates in the future.

Objective: To study the effect of the traditional Chinese therapy of tonifying the kidney and invigorating blood circulation (TKIB) on male infertility., Methods: Forty-two infertile males with abnormal DNA fragmentation index (DFI) were randomly allocated into a TKIB (n = 22) and a control group (n = 20), the former treated by TKIB with an oral Chinese medicinal prescription while the latter with oral tamoxifen tablets and vitamin E capsules, both for 3 months. Before and after treatment, we obtained the semen parameters and sperm DFI from the patients and followed them up for the outcomes of natural pregnancy., Results: Compared with the baseline, the patients in both the TKIB and control groups showed significant increases after medication in sperm concentration (［36.82 ± 29.16］ and ［34.56 ± 37.03］ vs ［50.00 ± 39.16］ and ［40.72 ± 47.37］ ×10⁶/ml, P<0.05), the percentage of progressively motile sperm (PMS) (［20.62 ± 9.10］% and ［21.25 ± 9.11］% vs ［36.82 ± 13.45］% and ［26.18 ± 10.60］%, P<0.05) and the percentage of morphologically normal sperm (MNS) (［1.28 ± 1.00］% and ［1.48 ± 0.91］% vs ［3.44 ± 1.33］% and ［2.57 ± 1.32］%, P<0.05), but remarkably decreased sperm DFI (［29.07 ± 11.52］% and ［24.43 ± 8.46］% vs ［15.51 ± 11.31］% and ［18.53 ± 10.44］%, P<0.05). The patients of the TKIB group exhibited an even higher total sperm motility and percentages of PMS and MNS than those of the control group (P<0.05) but no statistically significant difference from the latter in sperm concentration or DFI (P>0.05). Besides, the former achieved higher rates of natural pregnancy (18.2%) and live birth (18.2%) than the controls (15% and 10%) though neither with statistically significant difference (P>0.05)., Conclusions: The traditional Chinese therapy of tonifying the kidney and invigorating blood circulation can reduce sperm DNA damage and improve the outcomes of natural pregnancy in infertile men.

Objective: To investigate the antioxidative and spermatogenesis-repairing effects of Shenjing Guben Pills (SGP), a Chinese medicine for invigorating the kidney and blood circulation, on the testis, epididymis and sperm in rats with oxidative stress injury (OSI) induced by cadmium chloride., Methods: Seventy-two male Wistar rats were equally randomized into six groups: normal control, OSI model control, Wuzi Yanzong Pills (WYP) and low-, medium- and high-dose SGP. The OSI model was made in the latter five groups by intraperitoneal injection of cadmium chloride at 1 mg/kg, and 24 hours later, the rats of the normal and model control groups treated intragastrically with 0.9% normal saline, those of the WYP group with WYP at 4.5 g/kg/d, and those of the low-, medium- and high-dose SGP groups with SGP at 2.8, 5.6 and 11.2 g/kg/d, respectively, all for 56 days. Then, all the animals were sacrificed for obtainment of the visceral indexes and histopathological changes of the testis, epididymis and seminal vesicle, measurement of sperm concentration and motility and the percentage of morphologically normal sperm (MNS) in the epididymis, and determination of the levels of glutathione perox-idase (GSH-PX), superoxide dismutase (SOD), malondial-dehyde aldehyde (MDA) and serum testosterone (T)., Results: Compared with the OSI model controls, the rats in the high-, medium- and low-dose SGP groups showed significantly higher visceral indexes of the testis (［0.237 ± 0.098］ vs ［0.403 ± 0.090］, ［0.357 ± 0.150］ and ［0.348 ± 0.140］ g/100 g, P < 0.05) and seminal vesicle (［0.241 ± 0.118］ vs ［0.347 ± 0.115］, ［0.336 ± 0.090］ and ［0.320 ± 0.065］ g/100 g, P < 0.05) and those of the high-dose SGP group in the epididymal index (［0.099 ± 0.088］ vs ［0.156 ± 0.030］ g/100 g, P < 0.05). In comparison with the OSI model controls, the animals of the high-, medium- and low-dose SGP groups exhibited significant increases in sperm concentration (［10.5 ± 17.7］ vs ［58.1 ± 32.2］, ［36.0 ± 36.2］ and ［31.9 ± 32.7］ ×10⁶/ml, P < 0.05) and serum T (［2.56 ± 0.75］ vs ［3.62 ± 0.96］, ［3.48 ± 1.33］ and ［3.24 ± 0.83］ nmol/L, P < 0.05 or P < 0.01), and those of the high- and medium-dose SCG groups in total sperm motility (［9.5 ± 13.0］% vs ［26.5 ± 15.5］% and ［18.9 ± 8.2］%, P < 0.05) and MNS (［36.2 ± 40.2］% vs ［85.3 ± 23.3］% and ［65.8 ± 28.1］%, P < 0.05) and the levels GSH-PX (［3.62 ± 2.22］ vs ［5.70 ± 1.73］ and ［5.42 ± 2.35］ U/mg prot, P < 0.05 ) and SOD (［41.3 ± 8.8］ vs ［52.7 ± 14.6］ and ［51.3 ± 14.7］ U/mg prot, P < 0.05). The MDA level, however, was markedly decreased in the high-, medium- and low-dose SGP groups (［0.41 ± 0.29］, ［0.44 ± 0.19］ and ［0.47 ± 0.20］ nmol/mg prot) as compared with that in the OSI model controls (［0.69 ± 0.28］ nmol/mg prot) (P < 0.05). Histopathological examinations manifested coagulative necrosis, calcification and disappearance of spermatogenic and Sertoli cells in the seminiferous tubules of the OSI model controls, with decreased intraluminal secretions and atrophic epithelial papillae in the seminal vesicles and non-sperm cells in the narrowed lumens of the atrophic epididymis. With the increased dose of SGP, the proportion of normal seminiferous tubules was enlarged, the epithelia of the seminal vesicle became column-shaped again, and the epididymal lumens grew lager with more sperm cells, which indicated a dose-dependent therapeutic efficacy. Medium- and high-dose SGP achieved a significantly better effect than WYP., Conclusions: Shenjing Guben Pills can antagonize oxidative stress, elevate the levels of testicular antioxidant enzymes and serum T, repair pathological injury of the testis, epididymis and seminal vesicle, and improve semen quality and spermatogenic function.

Objective: To investigate the efficacy and safety of lycopene in the treatment of BPH with lower urinary tract symptoms (LUTS)., Methods: Totally, 127 BPH patients with LUTS were treated with oral lycopene tablets at 500 mg bid in the Department of Andrology of Jinling Hospital from January 2018 to January 2019. At 8 and 16 weeks of medication, we compared the IPSS, quality of life (QOL) score, prostate volume, PSA level, maximum urinary flow rate (Qmax), postvoid residual urine volume (PVR) and the incidence of adverse reactions before and after treatment., Results: A total of 120 patients completed the treatment. Compared with the baseline, significant improvement was observed after 8 weeks of medication in the IPSS (［18.42 ± 4.59］ vs ［14.13 ± 4.51］, P < 0.01) and QOL score (［4.34 ± 1.37］ vs ［3.14 ± 1.25］, P < 0.01), and even more significant at 16 weeks in the IPSS (［18.42 ± 4.59］ vs ［10.29 ± 3.61］, P< 0.01), QOL score (［4.34 ± 1.37］ vs ［2.17 ± 1.35］, P < 0.01), PSA (［3.87 ± 3.14］ vs ［2.90 ± 3.07］ μg/L, P < 0.01), Qmax (［10.62 ± 2.08］ vs ［14.15 ± 3.66］ ml/s, P < 0.01) and PVR (［35.88 ± 15.33］ vs ［18.36 ± 13.09］ ml, P < 0.01), but there was no statistically significant difference in the prostate volume before and after treatment (［39.85 ± 10.22］ vs ［38.16 ± 10.12］ ml, P > 0.05), and no adverse reactions in any of the patients., Conclusions: Lycopene has a good therapeutic effect on BPH with LUTS, which can significantly improve the patients' quality of life without causing adverse reactions.

Background: Immune thrombocytopenia (ITP) is an immune-mediated acquired autoimmune hemorrhagic disease. About one-third of patients are unresponsive to first-line therapies. Thalidomide (THD) as an immunomodulatory agent is now used to treat several autoimmune disorders. Therefore, we assessed the safety and efficacy of THD in corticosteroid-resistant or relapsed ITP patients, and preliminarily explore its mechanism., Methods: 50 newly-diagnosed ITP patients and 47 healthy volunteers were enrolled in this study. Additionally, 17 corticosteroid-resistant or relapsed ITP patients were recruited, with 7 cases in the rhTPO + THD group and 10 cases in the THD monotherapy group. Overall response rate at 6, 12, and 24 months were assessed. Levels of Neuropilin-1(NRP-1), regulatory T cells (Tregs) and regulatory B cells (Bregs) were detected., Results: Expression of NRP-1, Tregs and Bregs were reduced in newly-diagnosed ITP patients. In vitro, THD treatment upregulated expression of NRP-1and Tregs only in ITP patients. As for corticosteroid-resistant or relapsed ITP patients, overall response rate at 6, 12, and 24 months was 85.7%, 57.1% and 100% in the rhTPO + THD group and 60%, 75% and 83.3% in the THD group, respectively. Additionally, rhTPO plus THD or THD therapy significantly increased the levels of NRP-1, Tregs and Bregs in responders., Conclusions: Our study shows for the first time that NRP-1 is involved in the pathogenesis of ITP, THD could induce response in ITP patients by upregulating NRP-1 expression and restoring the proportion of Tregs and Bregs. THD might be served as a novel therapeutic agent in corticosteroid-resistant or relapsed ITP patients., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Erythermalgia is a rare cutaneous disease characterized by episodic attacks of burning pain, erythema, and increased temperature. It primarily involves the extremities, with possible extensions to the ears, face, neck, and scrotum; in rare instances, it may afflict the ears, face, or the scrotum alone. Although various medications alone or in different combinations have been tried with significant variations in response, no recommended therapeutics have been established until very recently. This report presents two case histories of a 20-year-old and a 46-year-old woman, respectively, who displayed intermittent facial erythema associated with warmth and pain during the episodes and who presented normal between episodes. Both had good response to combinative treatments of systemic medication and topical lidocaine compounds. The younger was disease-free after more than 4 years of follow-up, and the older had recurrence after stopping the treatment. Conclusion: Erythermalgia may involve the face alone, and combinative approaches may be of choice for its treatment. Topical lidocaine compounds are considered to be a good option for palliative treatment.

Male infertility (MI) is a complex multifactorial disease, and idiopathic infertility accounts for 30% of cases of MI. At present, the evidence for the effectiveness of empirical drugs is limited, and in vitro fertilization is costly and may increase the risk of birth defects and childhood cancers. Therefore, affected individuals may feel obliged to pursue natural remedies. Traditional Chinese medicine (TCM) may represent a useful option for infertile men. It has been demonstrated that TCM can regulate the hypothalamic-pituitary-testicular axis and boost the function of Sertoli cells and Leydig cells. TCM can also alleviate inflammation, prevent oxidative stress, reduce the DNA fragmentation index, and modulate the proliferation and apoptosis of germ cells. Furthermore, TCM can supply trace elements and vitamins, ameliorate the microcirculation of the testis, decrease the levels of serum anti-sperm antibody, and modify epigenetic markers. However, the evidence in favor of TCM is not compelling, which has hindered the development of TCM. This review attempts to elucidate the underlying therapeutic mechanisms of TCM. We also explore the advantages of TCM, differences between TCM and Western medicine, and problems in existing studies. Subsequently, we propose solutions to these problems and present perspectives for the future development of TCM., Competing Interests: The authors have no potential conflicts of interest to disclose., (Copyright © 2019 Korean Society for Sexual Medicine and Andrology.)

Resonance absorption mechanism-based metasurface absorbers can realize perfect optical absorption. Further, all-dielectric metasurface absorbers have more extensive applicability than metasurface absorbers that contain metal components. However, the absorption peaks of the all-dielectric metasurface absorbers reported to date are very sharp. In this work, we propose a broadband optical absorption all-dielectric metasurface, where a unit cell of this metasurface is composed of two coupled subwavelength semiconductor resonators arrayed in the direction of the wave vector and embedded in a low-index material. The results indicate that the peak absorption for more than 99% is achieved across a 60 nm bandwidth in the short-wavelength infrared region. This absorption bandwidth is three times that of a metasurface based on the conventional design scheme that consists of only a single layer of semiconductor resonators. Additionally, the coupled semiconductor resonator-based all-dielectric metasurface shows robust perfect absorption properties when the geometrical and material parameters-including the diameter, height, permittivity, and loss tangent of the resonator and the vertical and horizontal distances between the two centers of the coupled resonators-are varied over a wide range. With the convenience of use of existing semiconductor technologies in micro/nano-processing of the surface, this proposed broadband absorption all-dielectric metasurface offers a path toward realizing potential applications in numerous optical devices.