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.

Objective This study aimed to investigate the relationship between multiple inflammatory markers and poor outcome in patients with intracerebral hemorrhage,and to compare their predictive abilities.Methods We retrospectively analyzed the patients with intracerebral hemorrhage admitted to the Department of Neurology of the First Affiliated Hospital of Nanchang University from January 1,2015 to March 31,2023.According to the Modified Rankin Scale at 90 days after onset,the patients were divided into good outcome(mRS≤2 points)and poor outcome(mRS score≥3 points).Clinical information,laboratory examinations and follow-up data were compared between the two groups.Inflammatory markers include neutrophil to lymphocyte ratio(NLR),platelet to lymphocyte ratio(PLR),monocyte to high-density lipoprotein ratio(MHR),systemic inflammatory response index(SIRI),systemic immune-inflammation index(SII),white blood cell count to mean platelet volume ratio(WMR),lymphocyte to monocyte ratio(LMR),and systemic coagulation-inflammation index(SCI).Univariate and multivariate logistic regression analysis were used to analyze the predictors of poor prognosis after intracerebral hemorrhage,and receiver operating characteristic curve(ROC)was constructed to compare their predictive ability.Results A total of 510 patients with intracerebral hemorrhage were included.Of those,297(58.2%)had good outcome,and 213(41.8%)had poor outcome.Comparison of baseline characteristics demonstrated that patients with poor prognosis had higher levels of white blood cells,neutrophils,high-density lipoprotein,fibrinogen,NLR,PLR,SIRI,SII,WMR at admission,larger baseline hematoma volume and baseline perihematomal edema volume,a higher proportion of lobar hemorrhage,older age,and lower levels of platelets,lymphocytes,LMR,and SCI(P<0.05).Multivariate regression analysis showed that NLR(OR:1.081,95%CI:1.032～1.1131,P=0.001),SIRI(OR:1.089,95%CI:1.014～1.169,P=0.019),SII(OR:1.000,95%CI:1.000～1.001,P=0.011),WMR(OR:2.627,95%CI:1.267～5.445,P=0.009)were independently associated with poor prognosis in patients with ICH.In ROC analysis,the area under the curve of NLR(0.729,95%CI:0.685～0.774)was higher than SIRI(0.692,95%CI:0.645～0.738),SII(0.688,95%CI:0.641～0.735)and WMR(0.65,95%CI:0.602～0.698)for predicting poor outcomes.Conclusion NLR,SIRI,SII and WMR at admission are independently associated with poor outcomes in patients with intracerebral hemorrhage,and NLR has the strongest predictive ability.

AIM To evaluate the quality consistency of Tongmai Granules,Tongmai Tablets,Tongmai Capsules and Tongmai Oral Liquid.METHODS The HPLC fingerprints were established,after which the contents of danshensu,protocatechuic aldehyde,3'-hydroxy puerarin,puerarin,puerarin apioside,daidzin,ferulic acid,salvianolic acid B and salvianolic acid A were determined,and cluster analysis and principal component analysis were adopted in the quality analysis from the perspective of daily intake.RESULTS There were 21 common peaks in the fingerprints for 39 batches of samples with the similarities of 0.765-0.997.Various batches of samples were clustered into 5 categories,2 principal components demonstrated the accumulative variance contribution rate of 83.53％ .The daily intakes of various constituents in different dosage forms exhibited obvious differences,especially for that of salvianolic acid B,which were low in tablets and capsules,and their heterogeneities existed among the same dosage forms.CONCLUSION This simple and accurate method can provide a reference for the quality evaluation of Tongmai preparations from different manufacturers.

Aim To explore the effect of the classical famous prescription Dahuang Zhechong pill(DHZCP)on relieving liver cirrhosis by regulating the stress fiber remodeling mediated by mechanistic signaling pathway and to explore the underlying mechanism.Methods Mice were randomly divided into the control group,model group,DHZCP low-dose group,DHZCP high-dose group,and Colchicine-positive control group.The liver cirrhosis mouse model was constructed by intrap-eritoneal injection of olive oil-solubilized CCl4.HE staining and serologic markers were used to reflect liver injury.Masson staining was used to evaluate collagen deposition in liver tissue.ELISA was applied to detect vasoactive molecules and cancer indicators.Atomic force microscopy was employed to detect liver tissue stiffness.Color Doppler diagnostic instrument was used to assess portal blood flow velocity.Western blot was utilized to detect ROCK2 expression and phosphoryla-tion of YAP,Cofilin,and MLC.Results The liver tis-sues in the model group had obvious inflammatory cell infiltration and collagen deposition,accompanied by significant elevation of serum transaminases and fibrosis indexes.Similarly,vasoactive molecules and cancer in-dicators were elevated,and the mechanoregulatory pro-tein ROCK2 expression and phosphorylation of Cofilin and MLC were elevated,with YAP being strongly de-phosphorylated.Both low and high doses of DHZCP re-versed the pathological changes,serological indices,and inhibited the activation of the stress fiber(SF)re-modeling mechanistic signaling pathway.Conclusion DHZCP effectively ameliorates liver tissue lesions in mice with liver cirrhosis,and its mechanism may be re-lated to the inhibition of SF remodeling mechanistic signaling pathway.

Objective To observe the clinical effect of hysteroscopic combined with laparoscopic repair and reconstruction of diverticulum in the treatment of post-cesarean section uterine diverticulum(PCSD).Methods A total of 150 patients with PCSD admitted to the department of gynaecology of our hospital from January 2020 to February 2022 were selected and divided into the transvaginal group(transvaginal repair of diverticulum)and the combined group(hysteroscopic combined with laparoscopic repair and reconstruction of diverticulum)according to different surgical plans,with 75 cases in each group.The general condition of surgery,days of menstrual period,thickness of muscular layer,inflammation-related factor levels,complications and pregnancy in patients with birth plan were recorded in both groups.Results The operation time of patients in the combined group was longer than that in the transvaginal group,and the time of anal exhaustion and postoperative hospital stay of patients in the combined group were shorter than those in the transvaginal group,the differences were statistically significant(P<0.05);There was no significant difference in terms of the intraoperative bleeding volume of patients between the two groups(P>0.05).The high-sensitivity C-reactive protein(hs-CRP),interleukin-4(IL-4),interleukin-8(IL-8),and tumor necrosis factor-α(TNF-α)7 days after surgery of patients were lower in both groups compared with those before surgery,and the combined group was lower than the transvaginal group,the differences were statistically significant(P<0.05).Compared with that before the surgery,the days of menstrual period of patietns in both groups 6 and 12 months after the surgery decreased,and the thickness of muscular layer increased,with statistically significant differences(P<0.05);while no statistically significant difference was found in terms of the days of menstrual period or thickness of muscular layer of patients 6 and 12 months after surgery between the two groups(P>0.05).There was no significant difference in the incidence of cumulative complications such as incision infection,hyponatraemia,bladder injury and vaginal infection between the two groups(P>0.05).The intrauterine pregnancy rate of patients with birth plan and the full-term pregnancy rate in the combined group were higher than those in the transvaginal group,the differences were statistically significant(P<0.05).There was no statistically significant difference in the abortion rate or preterm birth rate between the two groups(P>0.05).Conclusion The hysteroscopic combined with laparoscopic repair and reconstruction of diverticulum for patients with PCSD has a fast postoperative recovery,a short hospital stay,and can increase the pregnancy rate of patients with birth plan.

Objective:To explore the clinical application value of multimodal Magnetic Resonance Imaging(MRI)in cognitive impairment related to Cerebral Small Vessel Disease(CSVD).Methods:This study was a retrospective analysis of 60 CSVD patients,who were divided into cognitive impairment group(n=32)and normal group(n=28)based on Montreal Cognitive Assessment scale(MoCA).Compare the general information and MRI parameters of two groups of patients,use correlation analysis and logistic regression analysis to investigate the relationship between MRI parameters and cognitive dysfunction,and use receiver operating characteristic curve(ROC)to explore the value of MRI parameters in diagnosing cognitive dysfunction in cerebral small vessel disease.Results:Compared with the cognitively normal group,the cognitive impairment group had older age,higher apparent diffusion coefficient(ADC),lower anisotropy score(FA),and an overall increase in microbleeds(P＜0.05).Age(r=-0.510),ADC value(r=-0.591),and overall number of microbleeds(r=-0.369)were significantly negatively correlated with MoCA score(P＜0.05),while FA value(r=0.262)was significantly positively correlated with MoCA score(P＜0.05).The logistic regression results indicate that age,overall microbleeds,ADC value,and FA value are influencing factors for cognitive impairment in patients with cerebral small vessel disease.ROC curve analysis showed that the combination of age,ADC,FA,and microbleeds significantly improved diagnostic efficacy(AUC=0.990,95％CI=0.974-1.000).Conclusion:The joint analysis of multimodal MRI parameters can provide important imaging evidence for the early identification of cognitive dysfunction in CSVD.

The aim of this study was to establish a rapid,highly sensitive,and specific nucleic acid detection method for the H5N6 and H9N2 avian influenza virus(AIV)subtypes by using reverse transcription-recombinase polymerase amplification(RT-RPA)combined with clustered regularly interspaced short palindromic repeats(CRISPR)-Cas13a proteins.The conserved regions were selected to design specific RT-RPA primers and crRNA sequences of H5,H6,H9,and N2 genes.RT-RPA tech-nology combined with CRISPR-Cas13a detection was used to evaluate the sensitivity and specificity of AIV nucleic acid detec-tion.The detection was performed on avian influenza environmental samples and compared with the results of fluorescence quantitative RT-PCR,to evaluate the effectiveness of the RT-RPA technology combined with CRISPR-Cas13a.AIV H5,H9,and N2 subtypes were detected with a sensitivity as high as 1 copy/μL,and AIV N6 subtypes were detected with a sensitivity of 10 copies/μL.Plasmid samples with differing copy numbers showed fluorescence under blue LED transillumination.The four AIV subtypes showed high specificity and did not cross-react with the other AIV subtypes.The detection of avian influenza ex-ternal environmental samples containing AIV H5,N6,and H9 subtypes was consistent with the results of fluorescence quanti-tative RT-PCR,with 100％accuracy.For AIV N2 subtypes,one additional negative sample was detected with 97.9％accuracy.The established RT-RPA technology combined with CRISPR-Cas13a detection enabled sensitive,specific visual detection of AIV H5N6 and H9N2 subtypes.This study provides a new nucleic acid detection method for AIV surveillance and subtype clas-sification.

The aim of this study was to establish a rapid,highly sensitive,and specific nucleic acid detection method for the H5N6 and H9N2 avian influenza virus(AIV)subtypes by using reverse transcription-recombinase polymerase amplification(RT-RPA)combined with clustered regularly interspaced short palindromic repeats(CRISPR)-Cas13a proteins.The conserved regions were selected to design specific RT-RPA primers and crRNA sequences of H5,H6,H9,and N2 genes.RT-RPA tech-nology combined with CRISPR-Cas13a detection was used to evaluate the sensitivity and specificity of AIV nucleic acid detec-tion.The detection was performed on avian influenza environmental samples and compared with the results of fluorescence quantitative RT-PCR,to evaluate the effectiveness of the RT-RPA technology combined with CRISPR-Cas13a.AIV H5,H9,and N2 subtypes were detected with a sensitivity as high as 1 copy/μL,and AIV N6 subtypes were detected with a sensitivity of 10 copies/μL.Plasmid samples with differing copy numbers showed fluorescence under blue LED transillumination.The four AIV subtypes showed high specificity and did not cross-react with the other AIV subtypes.The detection of avian influenza ex-ternal environmental samples containing AIV H5,N6,and H9 subtypes was consistent with the results of fluorescence quanti-tative RT-PCR,with 100％accuracy.For AIV N2 subtypes,one additional negative sample was detected with 97.9％accuracy.The established RT-RPA technology combined with CRISPR-Cas13a detection enabled sensitive,specific visual detection of AIV H5N6 and H9N2 subtypes.This study provides a new nucleic acid detection method for AIV surveillance and subtype clas-sification.

Objective:To compare the clinical outcomes and costs in arthroscopic repair of small and medium-sized supraspinatu tendon tears between double-pulley suture-bridge (DPSB) and traditional suture bridge (SB).Methods:A retrospective study was conducted at Department of Joint Surgery, The Second Hospital Affiliated to Hengyang Medical School to analyze the data of 26 patients with small and medium-sized supraspinatus tendon tears who had been treated by SB repair from May 2018 to December 2020 (SB group) and those of 35 patients with small and medium-sized supraspinatus tendon tears who had been treated by DPSB repair from January 2021 to December 2022 (DPSB group). There were 61 patients in the 2 groups, including 44 males and 17 females, with an age of (59.1±7.5) years. The left shoulder was affected in 26 patients and the right shoulder in 35 patients. The tear size was small in 25 cases and medium in 36 ones. The total number of anchors used, total anchoring costs, and surgical time were recorded and compared between the 2 groups. Visual analogue scale (VAS), American Shoulder and Elbow Surgeons (ASES) score, University of California Los Angeles (UCLA) score and shoulder range of motion were used to evaluate the clinical outcomes of the 2 groups before surgery and at the last follow-up. Comparisons were made within and between the 2 groups. Tendon integrity was assessed using MRI or ultrasound at 3, 6, 12 months or at the last follow-up.Results:There was no statistically significant difference in the preoperative general data between the 2 groups, indicating comparability ( P>0.05). DPSB and SB groups were followed up for (28.1±3.5) and (27.1±1.8) months, respectively. There was no statistically significant difference between DPSB group and SB group in surgical time or total number of anchors ( P>0.05). The total costs of anchoring in DPSB group [(6,028.6±173.4) yuan] were significantly lower than those in SB group [(13,257.1±554.2) yuan] ( P<0.05). At the last follow-up, the anterior flexion, abduction, external rotation and internal rotation of the shoulder, as well as VAS pain score, ASES score and UCLA score, were significantly better in both DPSB group and SB group than their preoperative values ( P<0.05), but there were no statistically significant differences between DPSB group and SB group ( P>0.05). There was no significant difference either in tendon retear between DPSB group (2 cases) and SB group (1 case) ( P>0.05). No such complication as wound infection or nerve damage was found in either group. Conclusions:In arthroscopic repair of small and medium-sized supraspinatu tendon tears, both DPSB and SB techniques can achieve satisfactory and comparable clinical outcomes. However, DPSB leads to lower total costs of anchoring.

AIM To evaluate the quality consistency of Tongmai Granules,Tongmai Tablets,Tongmai Capsules and Tongmai Oral Liquid.METHODS The HPLC fingerprints were established,after which the contents of danshensu,protocatechuic aldehyde,3'-hydroxy puerarin,puerarin,puerarin apioside,daidzin,ferulic acid,salvianolic acid B and salvianolic acid A were determined,and cluster analysis and principal component analysis were adopted in the quality analysis from the perspective of daily intake.RESULTS There were 21 common peaks in the fingerprints for 39 batches of samples with the similarities of 0.765-0.997.Various batches of samples were clustered into 5 categories,2 principal components demonstrated the accumulative variance contribution rate of 83.53％ .The daily intakes of various constituents in different dosage forms exhibited obvious differences,especially for that of salvianolic acid B,which were low in tablets and capsules,and their heterogeneities existed among the same dosage forms.CONCLUSION This simple and accurate method can provide a reference for the quality evaluation of Tongmai preparations from different manufacturers.