Multidrug-resistant (MDR) bacterial infections have emerged as a serious challenge of global public health crisis. The overuse and misuse of conventional antibiotics have dramatically accelerated the emergence, evolution and worldwide spread of drug-resistant bacterial strains, necessitating urgent exploration of novel antibacterial strategies. Bacteriophages serve as natural bacterial predators offering distinct advantages including high host specificity, autonomous self-replication capabilities and cost-effective large-scale production. However, wild-type phages present significant clinical limitations due to their narrow host ranges, susceptibility to rapid immune clearance and poor penetration of bacterial biofilms, which severely restrict their therapeutic applications. The convergence of synthetic biology, nanotechnology and advanced gene editing technologies has accelerated the development of engineered bacteriophage platforms, providing programmable, scalable and clinically translatable pathways to overcome these inherent biological constraints. Here, we systematically delineate four fundamental strategies for engineered bacteriophage development. Chemical modification utilizes reactive functional groups such as amino, carboxyl and thiol moieties on capsid proteins through esterification, amidation or click chemistry reactions to achieve precise drug conjugation and surface functionalization. In vivo editing encompasses ultraviolet or chemical mutagenesis for random mutation induction, homologous recombination for targeted genetic alterations, recombineering methodologies including electroporation-mediated bacteriophage recombination engineering, and CRISPR-Cas systems for precise genome editing to enable exact genetic reconstruction and host range reprogramming. In vitro synthesis leverages genome engineering platforms where intact phage genomes are transferred into yeast or host bacteria to facilitate highly efficient homologous recombination, enabling large DNA fragment assembly and cross-gene host range expansion without bacterial toxicity constraints. Directed evolution combines artificial selection through mutation library screening with rational design approaches involving chimeric receptor binding protein construction or site-specific mutagenesis, effectively balancing the discovery of unknown adaptive pathways with targeted host specificity modification. Moreover, we comprehensively discuss therapeutic applications across diverse clinical scenarios. Engineered bacteriophage effectively disrupt bacterial biofilms through sophisticated functionalized delivery platforms including nanozyme-conjugated phages, phage-liposome nanoconjugates and bio-responsive hydrogels, demonstrating significantly enhanced bactericidal efficiency compared to unmodified free phages. These bioengineered vectors attenuate bacterial virulence and resensitize pathogens to antibiotics by delivering CRISPR-Cas systems or base editors to disrupt critical virulence factors such as pili, capsule synthesis machineries and quorum sensing systems, or by inactivating antibiotic resistance determinants including beta-lactamase genes. As an intelligent nanomedicine delivery platform, engineered bacteriophage enable precise pathogen elimination an through photocatalytic reactive oxygen species generation, immunomodulatory interventions, or controlled release of antibacterial drugs. Furthermore, oral administration of engineered bacteriophage facilitates microbiota modulation, which selectively eliminate intestinal pathogens while preserve beneficial commensal microbiota, thereby restoring microbial community balance and preventing complications associated with dysbiosis. Finally, we critically analyze persistent challenges including host strain matching complexity, evolution of bacterial resistance mechanisms, pharmacokinetic optimization requirements, optimal administration route selection, large-scale production quality control standards and clinical dosing determination protocols. Through multidisciplinary integration of synthetic biology, infectious disease medicine and immunology, future translational medicine studies of bacteriophage should establish comprehensive technical platforms encompassing rapid phage screening, intelligent rational design, rigorous in vivo evaluation and standardized clinical validation processes, ultimately advancing engineered bacteriophage from laboratory innovations to clinically approved therapeutics for effectively combating MDR bacterial infections.

Alzheimer’s disease (AD) is a chronic neurodegenerative disorder that severely affects the health of the elderly, marked by its incurability, high prevalence, and extended latency period. The current approach to AD prevention and treatment emphasizes early detection and intervention, particularly during the pre-AD stage of mild cognitive impairment (MCI), which provides an optimal “window of opportunity” for intervention. Clinical detection methods for MCI, such as cerebrospinal fluid monitoring, genetic testing, and imaging diagnostics, are invasive and costly, limiting their broad clinical application. Speech, as a vital cognitive output, offers a new perspective and tool for computer-assisted analysis and screening of cognitive decline. This is because elderly individuals with cognitive decline exhibit distinct characteristics in semantic and audio information, such as reduced lexical richness, decreased speech coherence and conciseness, and declines in speech rate, voice rhythm, and hesitation rates. The objective presence of these semantic and audio characteristics lays the groundwork for computer-based screening of cognitive decline. Speech information is primarily sourced from databases or collected through tasks involving spontaneous speech, semantic fluency, and reading, followed by analysis using computer models. Spontaneous language tasks include dialogues/interviews, event descriptions, narrative recall, and picture descriptions. Semantic fluency tasks assess controlled retrieval of vocabulary items, requiring participants to extract information at the word level during lexical search. Reading tasks involve participants reading a passage aloud. Summarizing past research, the speech characteristics of the elderly can be divided into two major categories: semantic information and audio information. Semantic information focuses on the meaning of speech across different tasks, highlighting differences in vocabulary and text content in cognitive impairment. Overall, discourse pragmatic disorders in AD can be studied along three dimensions: cohesion, coherence, and conciseness. Cohesion mainly examines the use of vocabulary by participants, with a reduction in the use of nouns, pronouns, verbs, and adjectives in AD patients. Coherence assesses the ability of participants to maintain topics, with a decrease in the number of subordinate clauses in AD patients. Conciseness evaluates the information density of participants, with AD patients producing shorter texts with less information compared to normal elderly individuals. Audio information focuses on acoustic features that are difficult for the human ear to detect. There is a significant degradation in temporal parameters in the later stages of cognitive impairment; AD patients require more time to read the same paragraph, have longer vocalization times, and produce more pauses or silent parts in their spontaneous speech signals compared to normal individuals. Researchers have extracted audio and speech features, developing independent systems for each set of features, achieving an accuracy rate of 82% for both, which increases to 86% when both types of features are combined, demonstrating the advantage of integrating audio and speech information. Currently, deep learning and machine learning are the main methods used for information analysis. The overall diagnostic accuracy rate for AD exceeds 80%, and the diagnostic accuracy rate for MCI also exceeds 80%, indicating significant potential. Deep learning techniques require substantial data support, necessitating future expansion of database scale and continuous algorithm upgrades to transition from laboratory research to practical product implementation.

Objective To explore the efficacy of 4R technology combined with prone five-direction cer-vical muscle strength training for neck type cervical spondylosis.Methods A total of 112 patients with neck and shoulder pain who visited the Affiliated Shapingba Hospital of Chongqing University and the Shapingba District Traditional Chinese Medicine Hospital in Chongqing from January to November 2024 were selected as research subjects.They were randomly assigned using a random number table method into an observation group and a control group,with 56 patients in each group.The observation group received 4R technology com-bined with prone five-direction cervical muscle strength training,while the control group received conventional rehabilitation treatment.Both interventions lasted for 4 weeks.Differences in Visual Analogue Scale(VAS)scores,Neck Disability Index(NDI)scores,and cervical range of motion in flexion,extension,lateral bending,and rota-tion were assessed before treatment and at 1 and 6 months after treatment for both groups.Results At 1 and 6 months of treatment,VAS scores in both groups decreased compared to before treatment,with the observation group lower than the control group,and the difference was statistically significant(P<0.05).At 1 and 6 months of treatment,NDI scores in both groups decreased compared to before treatment,with the observation group lower than the control group,and the difference was statistically significant(P<0.05).At 1 and 6 months of treatment,cervical flexion,cervical extension,and cervical lateral flexion in both groups increased compared to before treatment,and cervical rotation in the observation group increased compared to before treatment.Cervical extension and cervical rotation in the observation group were greater than those in the con-trol group,and only at 6 months of treatment was cervical lateral flexion in the observation group greater than in the control group,with all differences statistically significant(P<0.05).Conclusion The 4R technology combined with prone five-direction cervical muscle strength training can effectively improve cervical-type cer-vical spondylosis.

Objective To establish a molecular method for the identification of Aucklandia lappa Decne.and its common adulterants,Inula helenium L.,based on tetra-primer amplification refractory mutation system(ARMS).Methods After the analysis and comparison of internal transcribed spacer 2(ITS2)sequence differences between Aucklandia lappa Decne.and Inula helenium L.,single-nucleotide polymorphism(SNP)sites were found to design specific primers for identification,the ARMS method was established to specifically amplify samples from different sources,and identification was carried out according to the size of specific amplification bands.Results The results showed that 170 bp and 271 bp specific bands could be amplified by Aucklandia lappa Decne.and Inula helenium L.,respectively,under the following conditions:annealing temperature 56℃,cycle number 25,primer concentration ratio of internal and external primers 1∶4,agarose gel concentration 1.5%and using 2×TSINGKE Master Mix(blue),the detection limit of adulterates in Aucklandia lappa Decne.and Inula helenium L.mixture sample was 5%.The 23 samples collected from the market were identified,results comparison was the same among ARMS detection,DNA barcoding,and plant classification.Conclusion The ARMS method established in this study can accurately identify Aucklandia lappa Decne.and Inula helenium L.,which has the characteristics of strong specificity and high sensitivity and simple operation,can provide basic research for solving the problem of admixture identification and technical support for the quality control of Aucklandia lappa Decne.medicinal materials.

Objective:To investigate the application effects of learning pyramid theory combined with constructivism in the standardized training of nurses in the department of gastroenterology.Methods:We randomly divided 104 gastroenterology nurses who received standardized training from October 2021 to October 2023 into observation group and control group, with 52 nurses in each group. The control group received conventional teaching, while the observation group received a combination of learning pyramid theory and constructivist teaching. The two groups were compared in terms of theoretical and practical scores, core competencies, self-learning abilities, and professional values scores before and after training. SPSS 23.0 was used to perform the t-test. Results:After training, the two groups showed improvements in all the tests; and the scores were all significantly higher in the observation group than in the control group: theoretical knowledge [(88.61±15.62) vs. (81.76±14.27)], skill practice [(82.91±15.17) vs. (76.40±13.82)], Competency Inventory for Registered Nurses scale score [(157.83±21.73) vs. (125.40±15.77)], self-learning ability score [(123.38±21.62) vs. (97.43±15.23)], and Nurse Professional Values scale score [(89.15±14.63) vs. (78.52±12.84)].Conclusions:The teaching model of learning pyramid theory and constructivism theory can enhance standardized training effectiveness for gastroenterology nurses.

Objective:To evaluate the clinically curative effect of intervention treatment of electronic bronchoscope in treating tracheobronchial tuberculosis at clinical activity stage.Methods:Sixty patients with tracheobronchial tuberculosis at clinical activity stage(type I,II,III and VI)who admitted to Liuzhou People's Hospital from September 2020 to September 2023 were selected,and they were divided into drug group(anti-tuberculosis drug treatment)and combination group(anti-tuberculosis drug treatment+interventional treatment with electronic bronchoscope)by the random number table method,with 30 cases in each group.The curative effects of the two groups were observed,and the negative conversion rate of sputum bacteria,clinical symptom scores(cough symptom,expectoration symptom)before and after treatment,Modified British Medical Research Council Dyspnea Scale(mMRC)score between two groups were compared,and the differences in indicators of pulmonary function such as forced expiratory volume in the first second(FEV1),forced vital capacity(FVC)and maximum voluntary ventilation(MVV)between the two groups also were compared.And then,the incidence of complications was calculated.Results:During the 1,2 and 3 months of follow-up,there were respectively 21 cases,27 cases and 30 cases occurred negative conversion of sputum bacteria in 30 patients of the combination group,and there were respectively 15 cases,18 cases and 23 cases occurred negative conversion of sputum bacteria in 30 patients of the drug group.At the 1st month of follow-up,the negative conversion rate of sputum bacteria in combination group was higher than that in drug group,while there was no statistically significant difference between the two groups(P＞0.05).At the 2nd and 3rd month of follow-up,the negative conversion rate of sputum bacteria in the combination group was higher than that in the drug group,and the differences were statistically significant(x2=7.200,7.925,P<0.05).The effective rate of treatment of the combination group was 100%,which was higher than 80%of the drug group,and the difference of that between two groups was significant(x2=6.667,P<0.05).After 2 months of treatment,the mMRC score,cough symptom score and expectoration symptom score of the combination group were all lower than those of the drug group,and the differences were statistically significant(t=3.504,3.950,3.530,P<0.05).The improvement effects of FEV1,FVC and MVV of the combination group were all better than those of the drug group,and the differences were statistically significant(t=6.626,4.966,4.097,P<0.05).There was no significant difference in the incidence of complications between the two groups(P>0.05).Conclusion:Anti-tuberculosis drug therapy combined with electronic bronchoscopy intervention has a good therapeutic effect in clinically active tracheobronchial tuberculosis.

This study investigates the mechanism by which Xintong Granules improve myocardial ischemia-reperfusion injury(MIRI) through the regulation of gut microbiota and their metabolites, specifically short-chain fatty acids(SCFAs). Rats were randomly divided based on body weight into the sham operation group, model group, low-dose Xintong Granules group(1.43 g·kg~(-1)·d~(-1)), medium-dose Xintong Granules group(2.86 g·kg~(-1)·d~(-1)), high-dose Xintong Granules group(5.72 g·kg~(-1)·d~(-1)), and metoprolol group(10 mg·kg~(-1)·d~(-1)). After 14 days of pre-administration, the MIRI rat model was established by ligating the left anterior descending coronary artery. The myocardial infarction area was assessed using the 2,3,5-triphenyltetrazolium chloride(TTC) staining method. Apoptosis in tissue cells was detected by the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL) assay. Pathological changes in myocardial cells and colonic tissue were observed using hematoxylin-eosin(HE) staining. The levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6), creatine kinase MB isoenzyme(CK-MB), and cardiac troponin T(cTnT) in rat serum were quantitatively measured using enzyme-linked immunosorbent assay(ELISA) kits. The activities of lactate dehydrogenase(LDH), creatine kinase(CK), and superoxide dismutase(SOD) in myocardial tissue, as well as the level of malondialdehyde(MDA), were determined using colorimetric assays. Gut microbiota composition was analyzed by 16S rDNA sequencing, and fecal SCFAs were quantified using gas chromatography-mass spectrometry(GC-MS). The results show that Xintong Granules significantly reduced the myocardial infarction area, suppressed cardiomyocyte apoptosis, and decreased serum levels of pro-inflammatory cytokines(TNF-α, IL-1β, and IL-6), myocardial injury markers(CK-MB, cTnT, LDH, and CK), and oxidative stress marker MDA. Additionally, Xintong Granules significantly improved intestinal inflammation in MIRI rats, regulated gut microbiota composition and diversity, and increased the levels of SCFAs(acetate, propionate, isobutyrate, etc.). In summary, Xintong Granules effectively alleviate MIRI symptoms. This study preliminarily confirms that Xintong Granules exert their inhibitory effects on MIRI by regulating gut microbiota imbalance and increasing SCFA levels.
Animals ; Gastrointestinal Microbiome/drug effects* ; Rats ; Male ; Myocardial Reperfusion Injury/genetics* ; Drugs, Chinese Herbal/administration & dosage* ; Rats, Sprague-Dawley ; Apoptosis/drug effects* ; Humans ; Tumor Necrosis Factor-alpha/metabolism* ; Interleukin-6/genetics* ; Malondialdehyde/metabolism*

OBJECTIVE: To investigate the relationship between angiotensin Ⅱ type 1 receptor autoantibody (AT1-AA) and semen parameters. Methods： The semen samples of 820 male patients who were treated in the Reproductive Medicine Center of Taiyuan Central Hospital from August 2022 to August 2023 were retrospectively analyzed. The levels of AT1-AA and Ang Ⅱ of semen were detected by ELISA, and the function of AT1-AA was detected by cardiomyocyte beating assay in suckling rats. The patients were divided into low group, median group and high group according to the OD values of AT1-AA. The differences in general data and semen parameters between different groups were analyzed. And the correlation between AT1-AA level and semen parameters in semen of all study subjects was analyzed by the method of Spearman analysis. And the relationships between AT1-AA OD value, Ang Ⅱ level and semen parameters in the AT1-AA high value group were analyzed as well. RESULTS: AT1-AA was present in semen with good function. There was no significant difference in the general data of patients in different AT1-AA levels (P>0.05). In the comparison of semen parameters among the groups with different levels of AT1-AA, there were differences in sperm concentration, PR concentration, NP％, and ALH among the three groups (P<0.05). And AT1-AA OD value was positively correlated with total sperm count, sperm concentration, PR concentration, and NP％, and negatively correlated with semen volume (P<0.05). In the AT1-AA high value group, the OD value of AT1-AA in semen was negatively correlated with inactive sperm, and positively correlated with total motility (［PR+NP］％), curve rate, mean path rate, and ALH. However, there was no correlation between the level of Ang Ⅱ in semen and semen parameters (P>0.05). CONCLUSION The presence of AT1-AA in semen may be associated with the promotion of sperm motility.
Male ; Humans ; Autoantibodies ; Sperm Motility ; Semen ; Retrospective Studies ; Receptor, Angiotensin, Type 1/immunology* ; Animals ; Rats ; Angiotensin II ; Adult ; Sperm Count ; Semen Analysis ; Receptor, Angiotensin, Type 2/immunology*

OBJECTIVE: To investigate the effect of acupuncture on advanced cancer patients by meta-analysis. METHODS: Nine databases (the Cochrane Central Register of Controlled Trials, MEDLINE, Web of Science, Embase, China National Knowledge Infrastructure, the Cumulative Index to Nursing and Allied Health Literature, Chinese Biomedical Literature Database, China Science and Technology Journal Database, and WanFang Data) were searched for randomized controlled trials (RCTs) on acupuncture in advanced cancer patients published from inception to February 13, 2023 and updated to June 1, 2023. Primary outcomes were quality of life (QOL), while secondary outcomes were pain, fatigue, and adverse events (side effects). Data synthesis was performed using RevMan V.5.3 to calculate pooled effect sizes. RoB-2 was used for the risk of bias, and the quality of evidence was assessed using the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) tool. RESULTS: Totally 17 RCTs involving 1,178 participants were included, 15 of which were pooled for meta-analysis. Most studies demonstrated some concern for the overall risk of bias. The pooled data indicated that acupuncture was associated with improved QOL [mean difference (MD)=6.67, 95% confidence interval (CI): 5.09 to 8.26], pain (MD=-1.18, 95% CI -2.28 to -0.08), and adverse events (risk ratio=0.30, 95% CI: 0.26 to 0.57) compared with control groups. Fatigue outcome was not included. Heterogeneity was substantial, and GRADE evidence was very low for both QOL and pain. CONCLUSIONS Acupuncture could benefit patients with advanced cancer and is considered safe compared with usual care. However, the evidence regarding QOL and pain outcomes requires further validation. It is crucial to encourage the development of high-quality studies to strengthen this evidence. (Registry No. CRD42023423539).
Humans ; Acupuncture Therapy ; Neoplasms/therapy* ; Quality of Life ; Randomized Controlled Trials as Topic ; Treatment Outcome

Gut microbial communities are likely remodeled in tandem with accumulated physiological decline during aging, yet there is limited understanding of gut microbiome variation in advanced age. Here, we performed a metagenomics-based enterotype analysis in a geographically homogeneous cohort of 367 enrolled Chinese individuals between the ages of 60 and 94 years, with the goal of characterizing the gut microbiome of elderly individuals and identifying factors linked to enterotype variations. In addition to two adult-like enterotypes dominated by Bacteroides (ET-Bacteroides) and Prevotella (ET-Prevotella), we identified a novel enterotype dominated by Escherichia (ET-Escherichia), whose prevalence increased in advanced age. Our data demonstrated that age explained more of the variance in the gut microbiome than previously identified factors such as type 2 diabetes mellitus (T2DM) or diet. We characterized the distinct taxonomic and functional profiles of ET-Escherichia, and found the strongest cohesion and highest robustness of the microbial co-occurrence network in this enterotype, as well as the lowest species diversity. In addition, we carried out a series of correlation analyses and co-abundance network analyses, which showed that several factors were likely linked to the overabundance of Escherichia members, including advanced age, vegetable intake, and fruit intake. Overall, our data revealed an enterotype variation characterized by Escherichia enrichment in the elderly population. Considering the different age distribution of each enterotype, these findings provide new insights into the changes that occur in the gut microbiome with age and highlight the importance of microbiome-based stratification of elderly individuals.
Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Bacteroides ; China ; Diabetes Mellitus, Type 2/microbiology* ; Escherichia coli/classification* ; Gastrointestinal Microbiome/genetics* ; Metagenomics ; East Asian People