Dental caries, a chronic disease characterized by tooth decay, occupies the second position in terms of disease burden and is primarily caused by cariogenic bacteria, especially Streptococcus mutans, because of its acidogenic, aciduric, and biofilm-forming capabilities. Developing novel targeted anti-virulence agents is always a focal point in caries control to overcome the limitations of conventional anti-virulence agents. The current study represents an up-to-date review of in silico approaches of drug design against dental caries, which have emerged more and more powerful complementary to biochemical attempts. Firstly, we categorize the in silico approaches into computer-aided drug design (CADD) and AI-assisted drug design (AIDD) and highlight the specific methods and models they contain respectively. Subsequently, we detail the design of anti-virulence drugs targeting single or multiple cariogenic virulence targets of S. mutans, such as glucosyltransferases (Gtfs), antigen I/II (AgI/II), sortase A (SrtA), the VicRK signal transduction system and superoxide dismutases (SODs). Finally, we outline the current opportunities and challenges encountered in this field to aid future endeavors and applications of CADD and AIDD in anti-virulence drug design.

Objective:To explore the construction of management system of dynamic adjustment for supply catalogue of medical consumables based on diagnosis related groups(DRG),so as to realize the fine supervision and management of rational and compliant use for medical consumables.Methods:The management system of dynamic adjustment for supply catalogue of medical consumables was constructed from 6 dimensions(system establishment,hierarchical management,detailed classification,announcement management,strengthening trial and standardizing contract)through analyzed the existing problems.A total of 149 broad headings of using medical consumables in clinical work of Children's Hospital,Zhejiang University School of Medicine from January 2022 to December 2023 were selected.The changes of the ratio of hygienic material's income in medical income,and the ratio of hygienic material's cost in medically overall cost were compared between before adopted the management system of dynamic adjustment for supply catalogue of medical consumables(2022)and after adopted that(2023).Results:After dynamic adjustment,32 broad headings(21.48%)of 149 broad headings of medical consumables were adjusted according to trial process,and 14 broad headings(9.40%)of medical consumables were optimized in the secondary catalogue,and 85 broad headings(57.05%)of medical consumables reduced their product brand,and the use of 8(5.37%)broad headings of medical consumables were stopped.On the premise that both the person-time of outpatient and emergency,and the person-time of discharge increased,the ratio of the income of hygienic material in medical income decreased from 8.02%of 2022 to 7.29%of 2023,and the ratio of hygienic material cost in overall medical cost decreased from 15.38%of 2022 to 14.17%of 2023,and the accumulated cost of medical consumables was reduced by 7.37 million CNY,accounting for 4.1%of the expenditure of medical consumables.Online procurement rates of medical consumables were respectively 93.05%and 94.34%in 2022 and 2023,which showed an increasing trend year by year.Conclusion:The application of management system of dynamic adjustment for supply catalogue of medical consumables can reduce the ratio of consumables of hospital,and improve the management efficiency of medical consumables,and reduce the procurement cost of medical consumables.

Objectives:To explore whether short-course radiotherapy (SCRT)-based total neoadjuvant therapy (TNT) combined with PD-1 inhibitors could further promote tumor regression and improve the prognosis.Methods:This is a prospective, multicenter, two-arm randomized controlled, seamless phase Ⅱ/Ⅲ trial for proficient mismatch repair or microsatellite stable (pMMR/MSS) locally advanced rectal cancer (LARC). Eligible patients were randomly assigned to the iTNT (TNT+PD-1) group or the TNT group. Patients in the TNT group received SCRT (5 Gy×5) followed by 4 cycles of CAPOX or 6 cycles of mFOLFOX chemotherapy, with the iTNT group receiving SCRT followed by the same regime in combination with 4 cycles of Sintilimab. Total mesorectal excision (TME) surgery or watch and wait (W&W) was performed after neoadjuvant therapy and then 2 cycles of same regimen as before were recommended. The primary endpoints are the complete response (CR) rate for phase Ⅱ trial and 3-year disease-free survival (DFS) for phase Ⅲ trial. A total of 588 patients will be enrolled for the phase Ⅱ/Ⅲ trial. Short-term efficacy and safety data from the initial 100 treated patients were analyzed as planned.Results:From 2022-8-31 to 2023-5-24 the initial 100 patients were enrolled from 10 hospitals in China, 76.0%(76/100) patients were male, and the median age was 61 years (21-74 years). More patients had tumors located in the lower rectum (78.0%, 78/100), staged T3-4 (97.0%, 97/100) and N1-2 (93.0%, 93/100), and about half of the tumors invaded the mesorectal fascia (52.0%, 52/100) and with extramural vascular invasion (51.0%, 51/100). Analyses were performed according to the per-protocal (PP) set. All patients in the iTNT group ( n=52) and the TNT group ( n=48) completed SCRT; The 4-cycle chemotherapy±Sintilimab completion rates were 86.5% and 100.0% in the iTNT and TNT groups, respectively. In the iTNT group, 82.7% (43/52), 11.5% (6/52), and 5.8% (3/52) of the patients received 4, 3, and 2 cycles of PD-1 inhibitor. After TNT, 68 patients underwent radical surgery and 15 patients achieved cCR and adopted W&W. The pathological complete response (pCR) rates were 48.5% (16/33) and 17.1% (6/35) in the iTNT and TNT groups, with CR rates of 50.0% (25/50) and 26.1% (12/46), respectively. The incidence of treatment-related grade 3-4 adverse events was 26.9% (14/52, iTNT group) and 18.8% (9/48, TNT group), with thrombocytopenia and leukopenia being the most common. Among patients receiving immunotherapy, grade 3 immunotherapy-related adverse events occurred in 2 (3.8%, 2/52) patients: one case was pancreatitis, another case was hepatitis combined with myositis and myocarditis. Conclusion:The preliminary results show that SCRT-based TNT combined with PD-1 inhibitors could further improve the CR rate for LARC without unexpected serious adverse events.

Dental caries,a chronic disease characterized by tooth decay,occupies the second position in terms of disease burden and is primarily caused by cariogenic bacteria,especially Streptococcus mutans,because of its acidogenic,aciduric,and biofilm-forming capabilities.Developing novel targeted anti-virulence agents is always a focal point in caries control to overcome the limitations of conventional anti-virulence agents.The current study represents an up-to-date review of in silico approaches of drug design against dental caries,which have emerged more and more powerful complementary to biochemical attempts.Firstly,we categorize the in silico approaches into computer-aided drug design(CADD)and AI-assisted drug design(AIDD)and highlight the specific methods and models they contain respectively.Subsequently,we detail the design of anti-virulence drugs targeting single or multiple cariogenic virulence targets of S.mutans,such as glucosyltransferases(Gtfs),antigen Ⅰ/Ⅱ(AgⅠ/Ⅱ),sortase A(SrtA),the VicRK signal transduction system and superoxide dismutases(SODs).Finally,we outline the current opportunities and challenges encountered in this field to aid future endeavors and applications of CADD and AIDD in anti-virulence drug design.

Iron overload refers to the pathological state in which the iron content in the body exceeds physiological requirements,Resultsing in the deposition of iron in the organs.Iron overload mouse models are an important tool for the study of iron metabolism disorders and related diseases.This paper summarizes the commonly used modeling method used in the construction of iron overload mouse models,which mainly comprise two categories.(1)exogenous iron overload mouse models,constructed through supplementation with extraenteral iron(injected iron)or intraintestinal iron(oral iron);(2)spontaneous iron overload models,constructed by screening for specific mouse lines or modifying iron metabolism-related genes.The method involving extraenteral iron supplementation has a short modeling duration and a high success rate,making it suitable for single and composite iron overload models.However,the high iron absorption rate may cause toxic reactions;thus,the optimal dose needs to be determined in advance.By contrast,intraintestinal iron supplementation is simple and safe,but has a low iron absorption rate and a long modeling duration.The spontaneous iron overload model is mainly used for specific genetic research studies,which are complex and involve high costs.The various modeling method offer diverse research pathways,spanning from molecular to systemic levels.This diversity is conducive to gaining an in-depth understanding of the pathological mechanisms underlying iron overload and provides an experimental basis for the development of new treatments.

Objectives:To explore whether short-course radiotherapy (SCRT)-based total neoadjuvant therapy (TNT) combined with PD-1 inhibitors could further promote tumor regression and improve the prognosis.Methods:This is a prospective, multicenter, two-arm randomized controlled, seamless phase Ⅱ/Ⅲ trial for proficient mismatch repair or microsatellite stable (pMMR/MSS) locally advanced rectal cancer (LARC). Eligible patients were randomly assigned to the iTNT (TNT+PD-1) group or the TNT group. Patients in the TNT group received SCRT (5 Gy×5) followed by 4 cycles of CAPOX or 6 cycles of mFOLFOX chemotherapy, with the iTNT group receiving SCRT followed by the same regime in combination with 4 cycles of Sintilimab. Total mesorectal excision (TME) surgery or watch and wait (W&W) was performed after neoadjuvant therapy and then 2 cycles of same regimen as before were recommended. The primary endpoints are the complete response (CR) rate for phase Ⅱ trial and 3-year disease-free survival (DFS) for phase Ⅲ trial. A total of 588 patients will be enrolled for the phase Ⅱ/Ⅲ trial. Short-term efficacy and safety data from the initial 100 treated patients were analyzed as planned.Results:From 2022-8-31 to 2023-5-24 the initial 100 patients were enrolled from 10 hospitals in China, 76.0%(76/100) patients were male, and the median age was 61 years (21-74 years). More patients had tumors located in the lower rectum (78.0%, 78/100), staged T3-4 (97.0%, 97/100) and N1-2 (93.0%, 93/100), and about half of the tumors invaded the mesorectal fascia (52.0%, 52/100) and with extramural vascular invasion (51.0%, 51/100). Analyses were performed according to the per-protocal (PP) set. All patients in the iTNT group ( n=52) and the TNT group ( n=48) completed SCRT; The 4-cycle chemotherapy±Sintilimab completion rates were 86.5% and 100.0% in the iTNT and TNT groups, respectively. In the iTNT group, 82.7% (43/52), 11.5% (6/52), and 5.8% (3/52) of the patients received 4, 3, and 2 cycles of PD-1 inhibitor. After TNT, 68 patients underwent radical surgery and 15 patients achieved cCR and adopted W&W. The pathological complete response (pCR) rates were 48.5% (16/33) and 17.1% (6/35) in the iTNT and TNT groups, with CR rates of 50.0% (25/50) and 26.1% (12/46), respectively. The incidence of treatment-related grade 3-4 adverse events was 26.9% (14/52, iTNT group) and 18.8% (9/48, TNT group), with thrombocytopenia and leukopenia being the most common. Among patients receiving immunotherapy, grade 3 immunotherapy-related adverse events occurred in 2 (3.8%, 2/52) patients: one case was pancreatitis, another case was hepatitis combined with myositis and myocarditis. Conclusion:The preliminary results show that SCRT-based TNT combined with PD-1 inhibitors could further improve the CR rate for LARC without unexpected serious adverse events.

Iron overload refers to the pathological state in which the iron content in the body exceeds physiological requirements,Resultsing in the deposition of iron in the organs.Iron overload mouse models are an important tool for the study of iron metabolism disorders and related diseases.This paper summarizes the commonly used modeling method used in the construction of iron overload mouse models,which mainly comprise two categories.(1)exogenous iron overload mouse models,constructed through supplementation with extraenteral iron(injected iron)or intraintestinal iron(oral iron);(2)spontaneous iron overload models,constructed by screening for specific mouse lines or modifying iron metabolism-related genes.The method involving extraenteral iron supplementation has a short modeling duration and a high success rate,making it suitable for single and composite iron overload models.However,the high iron absorption rate may cause toxic reactions;thus,the optimal dose needs to be determined in advance.By contrast,intraintestinal iron supplementation is simple and safe,but has a low iron absorption rate and a long modeling duration.The spontaneous iron overload model is mainly used for specific genetic research studies,which are complex and involve high costs.The various modeling method offer diverse research pathways,spanning from molecular to systemic levels.This diversity is conducive to gaining an in-depth understanding of the pathological mechanisms underlying iron overload and provides an experimental basis for the development of new treatments.

Objective:To explore the construction of management system of dynamic adjustment for supply catalogue of medical consumables based on diagnosis related groups(DRG),so as to realize the fine supervision and management of rational and compliant use for medical consumables.Methods:The management system of dynamic adjustment for supply catalogue of medical consumables was constructed from 6 dimensions(system establishment,hierarchical management,detailed classification,announcement management,strengthening trial and standardizing contract)through analyzed the existing problems.A total of 149 broad headings of using medical consumables in clinical work of Children's Hospital,Zhejiang University School of Medicine from January 2022 to December 2023 were selected.The changes of the ratio of hygienic material's income in medical income,and the ratio of hygienic material's cost in medically overall cost were compared between before adopted the management system of dynamic adjustment for supply catalogue of medical consumables(2022)and after adopted that(2023).Results:After dynamic adjustment,32 broad headings(21.48%)of 149 broad headings of medical consumables were adjusted according to trial process,and 14 broad headings(9.40%)of medical consumables were optimized in the secondary catalogue,and 85 broad headings(57.05%)of medical consumables reduced their product brand,and the use of 8(5.37%)broad headings of medical consumables were stopped.On the premise that both the person-time of outpatient and emergency,and the person-time of discharge increased,the ratio of the income of hygienic material in medical income decreased from 8.02%of 2022 to 7.29%of 2023,and the ratio of hygienic material cost in overall medical cost decreased from 15.38%of 2022 to 14.17%of 2023,and the accumulated cost of medical consumables was reduced by 7.37 million CNY,accounting for 4.1%of the expenditure of medical consumables.Online procurement rates of medical consumables were respectively 93.05%and 94.34%in 2022 and 2023,which showed an increasing trend year by year.Conclusion:The application of management system of dynamic adjustment for supply catalogue of medical consumables can reduce the ratio of consumables of hospital,and improve the management efficiency of medical consumables,and reduce the procurement cost of medical consumables.

Objective:To construct bacterial cytoplasmic membrane nanovesicles(BMV)with overexpressing programmed death 1(PD-1),denoted as BMV-PD-1 and evaluate the targeting efficacy of BMV-PD-1 towards transplanted lung tumor tissues in mice.Methods:The fusion plasmid ClyA-PD-1-EGFP fused by PD-1 and Cytolysin A(ClyA)was transferred into Escherichia coli BL21-Codonplus through plasmid transformation.Laser confocal microscopy,SDS-PAGE,and WB were used to detect the expression of the fusion protein ClyA-PD-1-EGFP.Bacterial membranes were extracted and processed with an extruder to generate BMV-PD-1.TEM and NTA were utilized to assess the morphology,size distribution,and zeta potential of BMV-PD-1,while WB was used to verify the presence of PD-1 protein.Laser confocal imaging was conducted to monitor the uptake of BMV-PD-1 by Lewis lung cancer cells.A C57BL/6J mouse subcutaneous transplant tumor model of LLC lung cancer cells was constructed,and the tumor targeting of BMV-PD-1 was evaluated by small animal imaging system.Results:Laser confocal microscopy images demonstrated that the plasmid ClyA-PD-1-EGFP was transferred into BL21-Codonplus and successfully expressed into protein.SDS-PAGE results suggested that ClyA-PD-1-EGFP was overexpressed in BL21-Codonplus.WB analysis indicated that PD-1 was expressed in bacteria and highly expressed in BMV-PD-1(P<0.001).NTA and TEM analyses revealed that BMV-PD-1 were spherical vesicles with a diameter of(145±14)nm and a negative surface charge.Laser confocal imaging showed that the high expression of PD-1 significantly increased the uptake of BMV-PD-1 by lung cancer cells(P<0.01).In vivo imaging of small animals further confirmed that the high expression of PD-1 can effectively improve cancer targeting of BMV-PD-1(P<0.01).Conclusion:In this study,bacterial plasma membrane nanovesicles BMV-PD-1 with high PD-1 expression are successfully constructed,and it is found that PD-1 overexpression markedly improve the mouse lung cancer xenograft tissue targeting specificity of BMV-PD-1,laying the groundwork for further development of BMV-PD-1 as a carrier for targeted drug delivery systems in tumors.

Specific tumor-targeted gene delivery remains an unsolved therapeutic issue due to aberrant vascularization in tumor microenvironment (TME). Some bacteria exhibit spontaneous chemotaxis toward the anaerobic and immune-suppressive TME, which makes them ideal natural vehicles for cancer gene therapy. Here, we conjugated ZIF-8 metal-organic frameworks encapsulating eukaryotic murine interleukin 2 (Il2) expression plasmid onto the surface of VNP20009, an attenuated Salmonella typhimurium strain with well-documented anti-cancer activity, and constructed a TME-targeted Il2 delivery system named Il2/ZIF-8@Salmonella. Both in vitro and in vivo experiments demonstrated that Il2/ZIF-8@Salmonella maintained the tumor-targeting feature of bacteria, and could be effectively phagocytosed by intratumoral macrophages, thus leading to the expression and secretion of IL2 in TME. The detailed analysis of tumor immune microenvironment (TIME) showed that one dose of combinatorial Il2/ZIF-8@Salmonella achieved synergistic actions on a potent remodeling of TIME, marked by the activation of cytotoxic T cells and M1-polarization of macrophages in TME, thus leading to significant anti-tumor effects in melanoma, orthotopic hepatocellular carcinoma, and pulmonary metastasis models. More importantly, Il2/ZIF-8@Salmonella exhibited high safety to major organs and hematopoietic systems. Taken together, we report a novel plasmid/ZIF-8@Salmonella system that simultaneously achieves effective TME-targeted delivery of therapeutic gene, as well as synergistic re-activation of TIME.