Objective: To investigate the incidence of occult hepatitis B virus infection (OBI) in spouses of asymptomatic hepatitis B virus (HBV) infected individuals in Shenzhen, China, and to analyze their serological and molecular characteristics and possible transmission routes, so as to propose refined strategies for blood safety. Methods: After rapid screening for HBsAg at the blood collection sites, samples from HBsAg-positive blood donors and their concurrently donating spouses were collected. All samples were tested for hepatitis B serological markers by electrochemiluminescence immunoassay (ECLI). Simultaneously, HBV nucleic acid extractiona, nested PCR amplification, gene sequencing of S and BCP/PC regions and qPCR were conducted. Results: A total of 112 samples were collected, including 56 from HBsAg positive donors and 56 from their spouses. All donors were confirmed as HBsAg+/DNA+/anti-HBc+, indicative of asymptomatic chronic hepatitis (CHB) infection. Among their 56 spouses, 11 (19.6%) were identified as HBV DNA+. The prevalence was higher in males (23.1%) than in females (16.7%). Six spouses (10.8%) had OBI, three of whom (5.4%) were negative in routine blood screening tests. The residual risk of HBV were estimated as 1∶127 (95%CI, 1∶356 to 1∶66). Among infected couples, immune escape mutation (E164D) and glycosylation mutations (I126T and T131N/M133T) were identified. Furthermore, sequence analysis suggested partner-to-partner transmission in eight cases. Conclusion: A substantial proportion (19.6%) of spouses of asymptomatic HBV infected donors were HBV-positve, with an OBI prevalence of 10.9%. Among these, 5.4% were negative in routine tests. To ensure blood safety, we recommend that spouses of HBV infected individuals be deferred from blood donation.

Coronary heart disease （CHD） has a distinct circadian rhythmicity in its onset， which is closely related to the waxing and waning of qi and blood in the channel passages of the human body. Grounded in the circadian rhythm theory of "correspondence between heaven and humankind"， this paper explores the pathogenic evolution rules of qi-blood disharmony， channel obstruction， interacting as cause and effect with circadian rhythm disorders in CHD. Clinically， CHD can be observed to attack in the early morning， at night， in the afternoon， and at fixed or unfixed circadian intervals. The pathogenesis rhythms at different time nodes can correspondingly reflect the susceptibility tendencies and pathological stress responses of the human body to core pathogeneses such as yang deficiency with cold congelation， yin deficiency with yang floating， qi and yin deficiency， and liver constraint with qi stagnation under specific temporal states. Treatment should be adapted to the season and time， regulating yin and yang in accordance with biological rhythms. It is recommended to regulate qi during the daytime by promoting ascending and dispersing to unblock the channel passages， and select the appropriate timing for administration to harness natural timing. At the Mao （卯） hour， yang ascends while yin descends， and taking medication at this time conforms to the ascending of yang qi. At the Wu （午） hour， the heart meridian is active， and taking medication at this time warms the channel passages and assists heart yang， which can be treated with modified Buzhong Yiqi Decoction （补中益气汤） or Shengyang Yiwei Decoction （升阳益胃汤）. At night， blood should be harmonized， when the channel passages are unblocked through nourishing， restraining， and searching， with Lianmei Decoction （连梅汤） combined with Dahuang Zhechong Pill （大黄䗪虫丸）. Following the temporal rhythm， the focus is put on the transitional time of day and night and the four seasons to treat disease before it arises， resonating and corresponding with the natural rhythm.

While pooled CRISPR screens have revolutionized functional genomics by enabling genome-scale interrogation, they have long faced a fundamental compatibility challenge with high-resolution microscopy-based phenotypes. The core issue lies in the technical disconnect between pooled screening formats, where cells are typically analyzed in bulk populations, and microscopy approaches that require spatial resolution and single-cell visualization. This limitation has created a significant blind spot in functional genomics, preventing researchers from systematically linking genetic perturbations to detailed subcellular architectural changes. To bridge this critical technological gap, a recent landmark study published in Developmental Cell has unveiled an integrated microscopy-based CRISPR screening platform that represents a paradigm shift in the field. The most innovative breakthrough of the platform involves extending photoactivation-based labeling techniques to fixed, immunostained cells, thereby overcoming a key limitation of prior approaches that were restricted exclusively to live-cell reporter systems. This methodological advancement allows researchers to capture transient cellular states and preserve delicate subcellular structures that would otherwise be lost in live-cell imaging scenarios. By seamlessly integrating this enhanced labeling approach with sophisticated automated imaging workflows and high-content analysis pipelines, the system enables unprecedented high-throughput, organelle-level phenotypic profiling at true single-cell resolution. The platform’s capabilities were demonstrated through its application to a comprehensive genome-wide screen for regulators of cilium assembly, a complex cellular process involving intricate cytoskeletal dynamics and membrane remodeling events. The screening results validated the platform’s robustness by successfully recapitulating known ciliary machinery components while simultaneously uncovering novel regulatory factors. Most notably, the study identified SMIM27/TZMP1, a conserved transition zone microprotein, as a previously unrecognized regulator of ciliary function. This discovery provides crucial insights into the molecular mechanisms governing ciliary transition zone barrier formation and selectivity. This accessible and versatile framework effectively overcomes longstanding bottlenecks in microscopy-based screening technologies, offering a generalizable strategy for dissecting organelle biology and cellular architecture at scale. The platform’s ability to combine genome-wide screening power with detailed morphological analysis opens new avenues for investigating cellular responses to genetic perturbations across diverse biological contexts.

Steroid-induced osteonecrosis of the femoral head （SONFH） is a severe musculoskeletal disorder often induced by the prolonged or excessive use of glucocorticoids. Characterized by ischemia of bone cells， necrosis， and trabecular fractures， SONFH is accompanied by pain， femoral head collapse， and joint dysfunction， which can lead to disability in severe cases. The pathogenesis of SONFH involves hormone-induced osteoblast apoptosis， bone microvascular endothelial cell （BMEC） apoptosis， oxidative stress， and inflammatory responses. The phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway plays a pivotal role in the development of the disease. Modulating the PI3K/Akt signaling pathway can promote Akt phosphorylation， thereby stimulating the osteogenic differentiation of bone marrow mesenchymal stem cells and osteoblasts， promoting angiogenesis in BMECs， and inhibiting osteoclastogenesis. The research on the treatment of SONFH with traditional Chinese medicine （TCM） has gained increasing attention. Recent studies have shown that TCM monomers and compounds have potential therapeutic effect on SONFH by intervening in the PI3K/Akt signaling pathway. These studies not only provide a scientific basis for the application of TCM in the treatment of SONFH but also offer new ideas for the development of new therapeutic strategies. This review summarized the progress in Chinese and international research on the PI3K/Akt signaling pathway in SONFH over the past five years. It involved the composition and transmission mechanisms of the signaling pathway， as well as its regulatory effects on osteoblasts， mesenchymal stem cells， osteoclasts， BMECs， and other cells. Additionally， the review explored the TCM understanding of SONFH and the application of TCM monomers and compounds in the intervention of the PI3K/Akt pathway. By systematically analyzing and organizing these research findings， this article aimed to provide references and point out directions for the clinical prevention and treatment of SONFH and promote further development of TCM in this field. With in-depth research on the PI3K/Akt pathway and the modern application of TCM， it is expected to bring safer and more effective treatment options for patients with SONFH.

Objective: To analyze the phylogenetic characteristics of VP1 gene of Coxsackievirus A10 (CVA10) isolates from 2004 to 2023, and to understand the genetic evolution and epidemic trends of CVA10, so as to provide references for the prevention and control of hand, foot, and mouth disease. Methods: The full-length sequences of the VP1 region of CVA10 isolates were retrieved from the BV-BRC database before December 15, 2024. Gene typing, sequence analysis, evolutionary analysis, and amino acid mutation site analysis were conducted using bioinformatics software. Results: A total of 1 253 CVA10 isolates VP1 region nucleotide full-length sequences from 2004 to 2023 were included, with 9 strains from 2004 to 2008, 338 strains from 2009 to 2012, and 906 strains from 2013 to 2023. China had the highest number of CVA10 isolates, with 1 143 strains accounting for 91.22%, and the predominant genotype was C3. Compared to the prototype strain, the nucleotide sequence homology of the VP1 region of CVA10 isolates ranged from 74.94% to 77.63%, while the amino acid sequence homology ranged from 88.59% to 93.62%. The third codon position preferred cytosine and thymine. The top three most abundant amino acids were threonine, alanine, and valine. The average relative synonymous codon usage of 30 amino acid codon groups was greater than 1. The average amino acid substitution entropy value was 0.04, with four amino acid mutation-prone sites identified, and the mutation-prone rate was 1.35%. Conclusions From 2004 to 2023, the majority of CVA10 isolates were primarily sourced from China, with genotype C3 being the predominant circulating strain in China. The nucleotide homology between the CVA10 isolates and the prototype strain was relatively low, and mutation-prone sites were identified, indicating that enhanced monitoring of viral variation is necessary.

OBJECTIVES: To explore the relationship between white matter injury (WMI) and cerebral perfusion in preterm infants using arterial spin labeling (ASL). METHODS: A total of 293 preterm infants (gestational age <34 weeks) hospitalized at the Third Affiliated Hospital of Zhengzhou University between June 2022 and June 2024 were included. After achieving clinical stability, the infants underwent brain magnetic resonance imaging (MRI) and ASL. Based on MRI findings, infants were classified into WMI (n=66) and non-WMI (n=227) groups. Cerebral perfusion parameters were compared between groups, and the association between WMI and perfusion alterations was evaluated. RESULTS: The WMI group showed a higher incidence of mild intraventricular hemorrhage (IVH) than the non-WMI group (P<0.05). Significantly lower cerebral perfusion was observed in the WMI group across bilateral frontal, temporal, parietal, and occipital lobes, as well as the basal ganglia and thalamus (P<0.05). After adjusting for gestational age, corrected gestational age at ASL scan, and mild IVH, WMI remained significantly associated with reduced regional perfusion (P<0.05). CONCLUSIONS WMI in preterm infants correlates with localized cerebral hypoperfusion. ASL-detected perfusion abnormalities may provide novel insights into WMI pathogenesis.
Humans ; White Matter/blood supply* ; Infant, Newborn ; Spin Labels ; Infant, Premature ; Female ; Male ; Cerebrovascular Circulation ; Magnetic Resonance Imaging

Objective:To investigate the clinical characteristics of patients with hepatitis B virus（HBV）-related primary hepatocellular carcinoma（HCC）who were treated in a multidisciplinary team（MDT）for liver cancer，so as to provide a basis for clinical optimization of the diagnosis and treatment of patients with chronic hepatitis B（CHB）.Methods:A retrospective analysis was performed for 482 HBV-related HCC patients who were treated with HCC-MDT every Thursday afternoon in The First Affiliated Hospital of the Army Medical University from January 2022 to May 2024，aged 18-87（55.54±10.84）years，86.93%（419/482）males and 13.07%（63/482）females. According to the different underlying liver diseases at the time of initial medical treatment and the different prognostic outcomes at the later follow-up，the differences in clinical characteristics between groups under different conditions were compared and analyzed，and the influencing factors of HCC prognosis were understood by Logistic regression analysis. Results:At the time of MDT presentation，the differences in HBeAg status（ χ2=6.311 ，P=0.043），γ-glutamyl traspeptidase（GGT）（ Z=6.277， P=0.043），alkaline phosphatase（ALP）（ Z=7.236 ，P=0.027），and model for end-stage liver disease（MELD）scores（ Z=6.111， P=0.047）among patients with different underlying liver diseases were statistically significant. At follow-up，6.75%（11/163）of HBV-related HCC patients who presented to MDT had a family history of HCC，and their cumulative mortality rate was as high as 60.8%（205/337）at least for 1 year. Mulitivariate Logistic regression analysis showed that different underlying liver disease at the time of initial medical treatment，HBV DNA replication level，MELD score and choice of anti-cancer treatment regimen were the influencing factors for the prognosis of HCC（all P<0.05）. The worse the degree of cirrhosis at the initial presentation，the higher the level of HBV DNA replication，and the higher the MELD score，the worse the prognosis for HCC. Conclusion:Advancing the diagnosis and treatment of CHB，maximizing the inhibition of HBV DNA replication，reducing the MELD score，and optimizing the anti-cancer treatment regimen can reduce the mortality rate of HBV-related HCC.

Turning to critical illness is a common stage of various diseases and injuries before death. Patients usually have complex health conditions, while the treatment process involves a wide range of content, along with high requirements for doctor′s professionalism and multi-specialty teamwork, as well as a great demand for time-sensitive treatments. However, this is not matched with critical care professionals and the current state of medical care in China. Telemedicine, which shortens the distance of medical professionals and the gap of disease diagnosis and treatments in various regions through electronic information, can effectively solve the current problem. Therefore, there is an urgent need to develop a standardized, high-quality visualization telemedicine round system .Therefore, experts have been organized to search domestic and foreign literature on telemedicine round for critically ill patients and to form this consensus based on clinical experiences so as to further improve the level of critical care treatments in regions.

Objective To investigate the protective effects of paeonol(PAE)on autophagy in human neuroblastoma cells(SH-SY5Y)induced by overexpression of α-synuclein(α-Syn),and to explore its related mechanism.Methods SH-SY5Y cells served as control group,while those induced with A53T-α-Syn mutation were used as model group.Additional groups included PAE(150 μg/ml)group,3-MA(1 mmol/L)group,and PAE(150 μg/ml)+3-MA(1 mmol/L)group.Cell viability was assessed using CCK-8 method,cell morphology was observed under an optical microscope,and protein expressions of α-Syn,LC3-Ⅱ,p62,Beclin-1,phosphorylated c-Jun N-terminal kinase(p-JNK),and p-Bcl-2 were determined by Western blotting.Results Compared with control group,model control exhibited decreased cell survival(P＜0.01),increased α-Syn expression(P＜0.001),reduced expression of autophagy-related proteins LC3-Ⅱ and Beclin-1(P＜0.01,P＜0.05),elevated autophagy substrate protein p62(P＜0.05),and decreased expression of autophagy pathway-related proteins p-JNK and Bcl-2(P＜0.05,P＜0.01).Compared with model group,PAE group showed increased cell survival(P＜0.01),decreased α-Syn and p62 protein expression(P＜0.01,P＜0.05),and increased expression of LC3-Ⅱ,Beclin-1,p-JNK and Bcl-2(P＜0.05).Compared with PAE group,3-MA+PAE group demonstrated increased α-Syn expression(P＜0.05).Conclusions PAE could attenuate the injury of SH-SY5Y cells induced by A53T-α-Syn and eliminate over-expressed α-Syn by activating autophagy pathway,which may be associated with the upregulation of JNK/Bcl-2 mediated autophagy pathway.

Objective To establish a stable,reliable,and clinically relevant porcine model of endotoxin-induced acute respiratory distress syndrome(ARDS).Methods Ten 8-month-old male Bama minipigs were deeply sedated,followed by invasive mechanical ventilation and electrocardiographic monitoring.Lipopolysaccharide(LPS)was intravenously pumped at 600 μg/(kg·h)for 3 hours,then maintained at 15 μg/(kg·h)thereafter.Dynamic monitoring was performed at five time points after LPS injection(LPS 0,1,3,5,and 8 h),including arterial blood gas analysis and chest computed tomography(CT)scans.Pathological examination of lung tissues obtained via bronchoscopic biopsy(HE staining and transmission electron microscopy)was conducted.These indicators were comprehensively used to evaluate the success of the animal model.Results At 5 hours after LPS administration,8 minipigs developed symptoms such as skin cyanosis,elevated body temperature,and respiratory distress.The oxygenation index decreased to<300 mmHg.Chest CT scans showed diffuse pulmonary infiltrates.Histopathology revealed alveolar edema and hyaline membrane formation.Transmission electron microscopy demonstrated disruption of pulmonary blood-air barrier,depletion of lamellar bodies in type Ⅱ pneumocytes,inflammatory cell infiltration,and exudation of plasma proteins and fibrin.Compared with LPS 0 h,at LPS 8 h,the oxygenation index and arterial blood pH were significantly decreased(P<0.001),while blood lactic acid and serum potassium were significantly increased(P<0.05);serum calcium and base excess were significantly decreased(P<0.05),and the lung injury score based on HE-stained lung sections was significantly increased(P<0.01).Conclusion The porcine ARDS model established by continuous LPS injection can dynamically simulate the pathophysiological characteristics and typical pathological manifestations of clinical septic ARDS,making it an effective tool to study the pathogenesis,prevention,and treatment strategies of septic ARDS.