Phages, as obligate bacterial and archaeal parasites, constitute a virus group of paramount ecological significance due to their exceptional abundance and genetic diversity. These biological entities serve as critical regulators in Earth's ecosystems, driving biogeochemical cycles, energy fluxes, and ecosystem services across terrestrial and marine environments. Within soil microbiomes, phages function as microbial "dark matter," maintaining the soil-plant system balance through precise modulation of the microbial community structure and functional dynamics. Despite the growing research interests in soil phages in recent years, the proportion of such studies in environmental virology remains disproportionately low, which is primarily attributed to researchers' limited familiarity with the research methodologies for phage microecology, incomplete technical frameworks, and inherent challenges posed by soil environmental complexity. To address these challenges, this review synthesizes cutting-edge methodologies for soil phage investigation from four aspects: (1) tangential flow filtration (TFF)-based phage enrichment strategies; (2) integrated quantification approaches combining double-layer agar plating, epifluorescence microscopy, and flow cytometry; (3) multi-omics analytical pipelines leveraging metagenomics and viromics datasets; and (4) computational frameworks merging machine learning algorithms with eco-evolutionary theory for deciphering phage-host interaction networks. Through comparative analysis of methodological principles, technical merits, and application scopes, we establish a comprehensive workflow for soil phage research. Future research in this field should prioritize: (1) construction of soil phage resource libraries, (2) exploration of RNA phages based on transcriptomes, (3) functional characterization of unknown genes, and (4) deep integration and interaction validation of multi-omics data. This systematic methodological synthesis provides critical technical references for addressing fundamental challenges in characterizing soil phages regarding the community structure, functional potential, and interaction mechanisms with hosts.
Bacteriophages/physiology* ; Soil Microbiology ; Ecosystem ; Microbiota ; Metagenomics/methods*

Objective To investigate the effect of pramlintide,a pancreatic amyloid peptide analog,on learning and memory of Alzheimer's disease(AD)mice through antioxidant stress,and to determine the expression of phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway.Methods The APP/PS1 mice were divided into a pramlintide treatment group(intraperitoneal injection of 0.5 μmol/L per day for 10 weeks)and an AD group(same dose of PBS),with 5 mice in each group.The learning and memory abilities were detected with water maze test,the pathological changes of the hippocampus were observed with HE staining and immunohistochemistry,the morphological characteristics of dendritic spines in hippocampus were observed after Golgi staining,and the ultrastructure of hippocampal neurons was observed through transmission electron microscopy(TEM).The content of malondialdehyde(MDA)and the level of superoxide dismutase(SOD)in the hippocampal tissue were detected by biochemical assay,and the levels of inflammatory factors IL-6,TNF-α and IL-1β were determined with ELISA.Western blotting was applied to measure the expression of PI3K/Akt signaling pathway related proteins in the hippocampus.In the cell experiment,SH-SY5Y cells were added with Aβ 1-42 to establish a cell model of AD.After the cells were treated with pramlintide,the levels of oxidative stress and inflammatory response were detected,and cell apoptosis was detected by immunofluorescence.Results The animal experiments showed that pramlintide treatment resulted in significantly shortened escape latency(P<0.01),increased platform crossings(P<0.01),and prolonged time to exploring hidden platform(P<0.01).In the hippocampal tissue of the pramlintide treatment group,HE staining displayed hippocampal neurons in high density and neat arrangement(P<0.05),immunohistochemical results showed significantly reduced Aβ protein(P<0.01),Golgi staining results demonstrated more dendritic spines(P<0.05),TEM revealed almost intact neuronal mitochondrial structure,with reduced vacuolization and clear and identifiable morphology.When compared with the AD group,the levels of oxidative stress and inflammatory response were decreased(P<0.01),and the relative expression of p-PI3K/PI3K and p-Akt/Akt proteins was increased(P<0.01)in the treatment group.In cell experiments,the levels of oxidative stress and inflammatory response were decreased in AD cell model after pramlintide treatment(P<0.01),and the results of immunofluorescence showed that cell apoptosis was declined(P<0.01).Conclusion Pramlintide can improve the cognitive function,reduce the hippocampal deposition of Aβ,reduce oxidative stress and inflammatory response,alleviate the pathological changes of neuronal ultrastructure,and enhance the expression of PI3K/Akt signaling pathway in AD mice.

Objective To investigate the expression level of circular RNA circ-Tns3 in Alzheimer's disease(AD)mice and its role in Aβ-induced neuronal damage.Methods Five APP/PS1 transgenic AD mice and 5 wild-type(WT)mice,weighting of 23～26 g and aged 6 months were subjected in the study.Morris water maze test was used to assess learning and memory abilities,and immunohistochemical staining was performed to observe the number of Aβ plaques in the hippocampal tissue.Subsequently,total RNA was extracted from the brains to detect the differential expression of circRNAs between AD and WT mice,and the results were further analyzed with Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis.The top 6 differentially expressed circRNAs were validated by real-time quantitative PCR(RT-qPCR).In in vitro experiments,Aβ1-42 was used to treat neuronal cells to establish AD cell model,and si-circ-Tns3 was transfected into Aβ1-42-treated neuronal cells to knock down circ-Tns3.RT-qPCR was used to determine the expression levels of circ-Tns3 and miR-671-5p.Cell viability and apoptotic rate were detected by CCK-8 assay and TUNEL staining,respectively.The levels of superoxide dismutase(SOD)and malondialdehyde(MDA)were measured using corresponding kits,and the levels of inflammatory factors IL-6,IL-1β,and TNF-α were detected with ELISA.The interaction between circ-Tns3 and miR-671-5p was verified by dual-luciferase reporter assay and RNA-binding protein immunoprecipitation(RIP)assay.The expression level of Sirt1 protein was detected by Western blotting.Results The 6-month-old AD mice exhibited significant cognitive impairment and Aβ deposition(P<0.01).There were 269 differentially expressed circRNAs identified between AD and WT mice,of which 159 were up-regulated and 110 down-regulated.GO and KEGG enrichment analyses showed that these differentially expressed circRNAs were mainly involved in synaptic transmission,memory,and cholinergic synapse signaling pathways.The expression of circ-Tns3 was significantly increased not only in the brain tissue of AD mice but also in neuronal cells after Aβ1-42 treatment.In cellular experiments,knockdown of circ-Tns3 significantly reduced cell viability and number of apoptotic cells in Aβ1-42-treated neuronal cells,decreased MDA content,increased SOD activity,and reduced the levels of IL-6,IL-1β,and TNF-α(P<0.01).The starBase database predicted that circ-Tns3 and miR-671-5p have complementary sequences,and dual-luciferase reporter and RIP assays confirmed their interaction.The bioinformatics database predicted that miR-671-5p and sirt1 have complementary sequences.Western blotting indicated that in neuronal cells treated with Aβ1-42,the expression of sirt1 was increased after knockdown of circ-Tns3(P<0.01).In Aβ1-42-treated neuronal cells,after knockdown of circ-Tns3,addition of miR-671-5p inhibitor significantly decreased the expression level of sirt1 protein(P<0.01).Conclusions circ-Tns3 is highly expressed in AD mice and cell model of AD.Knocking circ-Tns3 down improves neuronal damage.circ-Tns3 may be involved in the neuronal damage through regulating sirt 1 protein by binding to miR-671-5p.

Objective To investigate the state of rotator cuff injury in flight crew and to explore the effectiveness of extracorporeal shock wave therapy combined with functional exercise in treating rotator cuff injury.Methods A total of 60 flight crew with rotator cuff injuries in Dalian Rehabilitation Center from June 2022 to June 2023 were enrolled in this study.They were randomly assigned to experimental group(shock wave therapy combined with functional exercise)or the control group(functional exercise).The clinical efficacy was compared between the two groups after 4 weeks of treatment.Results All the research objects were male.There was no significant difference in the general information,such as age,flight duration,or course of the disease between the two groups(P>0.05).The visual analogue scale(VAS)scores were decreased,and Constant-Murley scores(CMS)and range of motion(ROM)were increased in both groups after treatment.The above-mentioned indexes in the experimental group were better than those in the control group(P<0.001).Conclusion The shock wave therapy and functional exercise has a more significant therapeutic effect on rotator cuff injuries than functional exercise alone.

[Objective]To explore the diagnostic value of conventional ultrasound combined with shear wave elastography (SWE) for sarcopenia in patients with chronic kidney disease (CKD).[Methods]The study included 94 CKD patients (34 with sarcopenia and 60 without). All patient underwent the Simplified Assessment Rating Questionnaire (SARC-CalF),Mini Nutritional Assessment (MNA),Short Physical Performance Battery (SPPB),grip strength test,bioelectrical impedance analysis (BIA),conventional muscle ultrasound and SWE of the thighs. We then compared the differences in indicators between the sarcopenia group and non-sarcopenia group,used Spearman correlation analysis to assess the relationship between the two examinations (conventional ultrasound and SWE) and other clinical indicators,identified the diagnostic markers for sarcopenia,created receiver operating characteristic (ROC) curves,calculated the area under the curve (AUC) and determined the diagnostic performance of conventional ultrasound,SWE and their combination. Binary logistic regression was used to analyze the influencing factors of sarcopenia in CKD patients and a combined diagnosis model was established.[Results]The sarcopenia group showed lower upper arm circumference,calf circumference,6-meter walking speed and handgrip strength than non-sarcopenia group,and the differences were statistically significant (P<0.05). The sarcopenia group exhibited lower SARC-CalF and SPBB scores,as well as more compromised nutritional status. Statistically significant differences were observed in the ultrasound parameters between the two groups,including thickness of the subcutaneous fat and rectus femoris,combined thickness of the rectus femoris and vastus intermedius,rectus femoris cross-sectional area,elastic modulus of the rectus femoris and vastus medialis (all P<0.05). The muscle mass index had a moderate positive correlation with muscle thickness and cross-sectional area of the rectus femoris (0.3

Objective To investigate the current status,research hotspots,and emerging trends in the field of TCM treatment for tic disorder(TD);To provide references for relevant research.Methods Literature on the TCM treatment for TD was retrieved from CNKI,VIP,Wanfang Data,Web of Science Core Collection,and PubMed databases from January 1,2000 to May 1,2022.NoteExpress 3.7.0.9296 software was used for literature management.VOSviewer 1.6.18 software was utilized to conduct co-occurrence analysis on author,institution,and keyword information.R 4.1.3 was applied to statistically analyze keyword frequency and time distributions and generate visualizations.Results A total of 1 520 articles were included in the analysis,involving 107 core authors.High-frequency keywords included experience of famous doctors,clinical observation,acupuncture,and ear acupoints.Commonly used Chinese herbal medicines included Acori Tatarinowii Rhizoma and Bombyx Batryticatus.Frequently prescribed TCM formulae included Wendan Decoction,Changpu Yujin Decoction,Yinqiao Powder,and Tianma Gouteng Decoction.Conclusion Current research hotspots in this field focus on experience of famous doctors,the clinical efficacy of TCM interventions for TD,and molecular biological mechanism studies.Emerging research trends include external TCM treatment and comorbidity investigations.

Children and adults differ significantly in physiology,biochemistry and immune function,which leads to sig-nificant differences in blood transfusion strategies between children and adults.To guide the clinical transfusion practice of pediatric patients and improve the prognosis of children,the National Health Commission organized the formulation and re-lease of the health industry standard Guideline for Pediatric Transfusion(WS/T 795-2022).This paper will briefly introduce some concepts that help understand of the Standard and the preparation process of the Standard,and explain and interpret the preparation of the"scope","general provisions"and"factors to consider"of the Standard,hoping to contribute to the understanding and implementation of the Standard.

Objective:To explore the clinical value of a trinity strategy for the treatment of multiple orthopedic trauma.Methods:A retrospective case series study was conducted to analyze the clinical data of 1 267 patients with multiple orthopedic trauma admitted to Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine and the First Affiliated Hospital of Navy Medical University from June 2013 to May 2023, including 862 males and 405 females, aged 18-93 years [(55.2±19.8)years]. Associated injuries included hemorrhagic shock in 632 patients, traumatic wet lung in 274, cranial injuries in 135, abdominal and pelvic bleeding in 116, pneumothorax in 89, urinary injury in 13, and vesical rupture in 8. All the patients were treated with the trinity strategy and the treatment process was divided into the phases of first aid, remodeling, and rehabilitation. The first aid phase focused on stabilizing symptoms and saving lives; the remodeling phase centered on restoring the anatomical structure and alignment; the rehabilitation phase aimed for functional recovery through the integration of both Western and traditional Chinese medicine. The all-cause mortality within 30 days after surgery and fracture healing time were calculated; the excellent and good rates of Constant-Murley shoulder score, Mayo elbow score, Gartland-Werley wrist score, Harris hip score, Hospital for Special Surgery (HSS) knee score and the American Orthopedic Foot & Ankle Society (AOFAS) ankle-hindfoot score at the last follow-up and the overall excellent and good rate of all joint function scores were measured. The short form health survey (SF-36) scores were collected preoperatively and at 6 months postoperatively, including 8 aspects such as physical functioning, physical role, bodily pain, general health, vitality, social functioning, emotional role, and mental health. The incidence of postoperative complications was recorded.Results:All the patients were followed up for 6-18 months [(10.2±4.2)months]. The mortality rate during the acute phase (within 30 days after surgery) was 2.37% with 12 deaths due to hemorrhagic shock, 10 due to traumatic brain injury, 6 due to multiple organ dysfunction syndrome (MODS), and 2 due to pulmonary infection. The average fracture healing time averaged 3.8-18 months [(11.5±4.2)months], with 89.49% of the patients having bone union within 12 months after surgery, 8.93% having bone union within 18 months after surgery, and 1.58% undergoing reoperation. For the patients with internal fixation failure and nonunion, the average healing time was extended to (10.2±2.2)months and (13.7±3.3)months respectively. At the last follow-up, the excellent and good rates of Constant-Murley shoulder score, Mayo elbow score, Gartland-Werley wrist score, Harris hip score, HSS knee score, and AOFAS ankle-hindfoot score were 83.93%, 90.24%, 94.12%, 85.57%, 88.46%, and 92.31% respectively, with an overall excellent and good rate of 89.11%. At 6 months after surgery, the SF-36 scores of all the patients in the eight dimensions,including the physical functioning, physical role, bodily pain, general health, vitality, social functioning, emotional role, and mental health were (74.4±8.6)points, (44.7±14.4)points, (77.4±10.9)points, (68.4±18.2)points, (72.5±16.0)points, (76.8±8.7)points, (49.9±17.6)points, and (72.8±17.9)points, significantly improved compared with those before operation [(63.4±12.7)points, (30.9±17.4)points, (56.4±18.0)points, (55.4±24.7)points, (53.5±21.0)points, (55.8±24.3)points, (36.9±24.0)points, (58.8±21.6)points] ( P<0.01). Complications of different degrees occurred in 214 patients (16.89%), including lung infections in 118 patients (9.31%), lower extremity deep vein thrombosis in 50(3.95%), pressure injuries in 26(2.05%), internal fixation failure in 12(0.95%), and nonunion in 8(0.63%). Conclusions:The trinity strategy provides whole-process management, personalized treatment, and overall rehabilitation for multiple orthopedic trauma. It can decrease mortality, shorten fracture healing time, improve joint function and quality of life, and reduce the incidence of complications.

Objective A scoping review of research on mobile glucose management in pregnant women with gestational diabetes mellitus was conducted to provide references for clinical providers to conduct mobile glucose management and related research.Methods Based on Arksey and O'Malley's scoping review reporting framework,JBI Library,Cochrane Library,PubMed,Web of Science,Embase,CINAHL Complete,CBM,China National Knowledge In-frastructure(CNKI),and Wanfang Database were retrieved.The search time frame was from the establishment of the databases to February 14,2023.The included literature was screened,summarized and analyzed.Results A total of 65 publications from 11 countries were included,among which 38 were randomized controlled trials,27 were experimental studies.The vehicles relied on for mobile glucose management in gestational diabetes mainly included APPs,remote monitoring systems,professional websites,and social platforms,which could provide health education,glucose management monitoring and recording,social support,and glucose management follow-up and reminders to pregnant women.Most of the studies focus on the glucose index and drug use,pregnancy outcome,self-management,quality of life,user experience and resource cost of pregnant women.Conclusion Mobile glucose management carriers are geographically specific and have a positive effect on glucose control in gestational diabetes.Future studies need to further develop localized mobile glucose management intervention protocols and conduct more high-quality randomized controlled studies to verify their effectiveness.

Objective To determine the role of programmed cell death-ligand l(PD-L1)and cluster of differentiation 8(CD8)in judging the prognosis of human epidermal growth factor receptor 2(HER2)positive breast cancer patients,through evaluating the expression of PD-L1 and CD8 in the tumor microenvironment,and their correlation with clinicopathological data.Methods A total of 65 HER2 positive breast cancer patients were collected admitted to Tengzhou Central People's Hospital in Shandong Province from October 2016 to October 2018,all of whom were women.Theparaffin slices of cancer tissue were labeled with PD-L1 and CD8 using immunohistochemical methods.Collect clinicopathological data of patients,including 7 prognostic parameters,such as age,histological grade,P53,KI-67,estrogen receptor,progesterone receptor and tumor node metastasis classification(TNM)staging.Statistical analysis was conducted to determine whether the expression of PD-L1 and CD8 were correlated with the above clinicopathological data.Results PD-L1 positive rate was 21.5％,the positive expression of PD-L1 was correlated with histological grading and TNM staging,and the difference in results was statistically significant(x2=6.250,P=0.044;x2=13.730,P=0.001);CD8 positive rate was 32.3％,the expression of CD8 cells was correlated with histological grading and TNM staging,and the difference in results was statistically significant(x2=8.023,P=0.018;x2=6.117,P=0.047).Conclusion PD-L1 and CD8 expression are correlated with breast cancer histological grade and TNM stage.PD-L1 and CD8 expression may be important prognostic markers for HER2 positive breast cancer patients.