Immunomodulatory cancer therapy is witnessing the rise of viral immunotherapy. The oncolytic influenza A virus, although promising in preclinical investigations, remains to be implemented in clinical practice. Recent progress in genetic engineering, coupled with experiential insights, offers opportunities to enhance the therapeutic efficacy of the influenza A virus. This review explores the use of the influenza virus, its attenuated forms, and associated vaccines in cancer immunotherapy, highlighting their respective advantages and challenges. We further elucidate methods for engineering influenza viruses and innovative approaches to augment them with cytokines or immune checkpoint inhibitors, aiming to maximize their clinical impact. Our goal is to provide insights essential for refining influenza A virus-based viral tumor immunotherapies.
Humans ; Neoplasms/immunology* ; Immunotherapy/trends* ; Influenza A virus/immunology* ; Oncolytic Virotherapy/trends* ; Animals ; Cancer Vaccines/therapeutic use* ; Oncolytic Viruses ; Genetic Engineering ; Immune Checkpoint Inhibitors/therapeutic use*

Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering, which limit sequence and functional diversity, thereby constraining their therapeutic and application potential. Protein design tools have significantly advanced the creation of novel protein sequences, structures, and functions. However, further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability. Here, we extended an evolution-based design protocol to create a novel minibinder, BindHer, against the human epidermal growth factor receptor 2 (HER2). It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity. Radiolabeling experiments with ^99mTc, ⁶⁸Ga, and ¹⁸F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models, with minimal nonspecific liver absorption, outperforming scaffolds designed through traditional engineering. These findings highlight a new rational approach to automated protein design, offering significant potential for large-scale applications in therapeutic mini-protein development.

Sarin(GB)is a typical representative of nerve agents with high toxicity,and very low amount can cause death.GB can cause water and atmospheric environment poisoning,so the detection of GB in water and air is of great significance.In this work,a colorimetric sensor array(CSA)based on GB inhibition of cholinesterase activity was constructed to detect GB with high selectivity.A 4×4 colorimetric array was constructed using acetylcholinesterase(AChE),butyryl cholinesterase(BuChE)and the corresponding substrate acetylthiocholine iodide(S-ACh),butyryl thiocholine iodide(S-BCh),acetylcholine chloride(ACh),butyryl choline chloride(BCh)and 2,6-dichloroindophenol ethyl ester(DCIE).The linear curve of the sensor was Y=131.3×lgC+271.6(R2=0.997),where Y was the array response Euclidean distance,C was the concentration of GB(mg/L),the linear range was 0.03?0.32 mg/L,and the detection limit was 27.6 μg/L.The method could effectively distinguish chemical warfare agents(CWA)such as VX,Soman(GD),mustard gas(HD),Louie reagent(L),and had high anti-interference ability,sensitivity and good repeatability.It was successfully applied to the detection of GB in simulated water and simulated air samples,and the sample recovery rate was 97.2% ?100.9%.This method would be potentially applied to the field rapid detection of nerve agents.

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.

AIM To investigate the mechanism of Shaoyao Gancao Decoction in preventing meglumine diatrizoate-induced post-ERCP pancreatitis in rats through autophagy regulation.METHODS The rats were randomized into the normal group,the model group,the low-dose and high-dose Shaoyao Gancao Decoction(1.5,3.0 g/kg),and the indomethacin suppository group.A rat model of post-ERCP pancreatitis was induced by meglumine diatrizoate injection into the pancreatic duct under continuous pressure.The rats had their pancreatic tissues stained with HE to observe the pathological alterations,inflammatory cell infiltration,hemorrhage and necrosis;their serum levels of IL-1β,IL-6,IL-8,TNF-α,AMS,and IL-10 identified by ELISA;their autophagic vacuoles in pancreatic acinar cells observed by transmission electron microscopy;their pancreatic protein expressions of Beclin1,LC3B,p62,TRAF2 and p-JNK detected by IHC and Western blot;and their pancreatic mRNA expressions of Beclin1 and TRAF2 detected by RT-qPCR.RESULTS Compared with the model group,the high-dose Shaoyao Gancao Decoction group displayed no obvious hemorrhage;improvement in edema of acinar and interstitial cells;obviously less cellular inflammatory infiltration;substantially decreased serum levels of IL-1β,IL-6,TNF-α and AMS(P＜0.05,P＜0.01);drastically reduced amount of autophagosomes in acinar cells;and down-regulated expressions of autophagy-related proteins Beclin1,LC3,p62,TRAF2 and p-JNK(P＜0.05,P＜0.01).CONCLUSION Shaoyao Gancao Decoction can prevent post-ERCP pancreatitis by ameliorating pancreatic tissue injury,decreasing serum inflammatory response level,and interfering with abnormal autophagy of pancreatic acinar cells.Its molecular mechanism may involve inhibition of TRAF2 protein expression and modulation of p-JNK activation.

Aim To study the effects of whole herb of Prunella and its active components on the malignant progression of breast cancer and its mechanism.Meth-ods Breast cancer transplantation tumor model was constructed and randomly divided into the model group,low,medium and high dose group of whole herb of Prunella(0.1,0.2,0.4 g·mL-1 by gavage)and paclitaxel(10 mg·kg-1 by intraperitoneal injection),which was administered by gavage every day,and the tumor tissues were collected after 28 days of interven-tion.The weight,tumor volume and mass of nude mice in each group were detected,HE staining was used to observe the morphology of breast cancer tumor tissues,and immunohistochemical staining was used to observe the proliferation of cell-cycle regulatory protein-67(Ki-67)and cytokeratin 17(CK17)in breast cancer tumor tissues.The cellular experiments were performed by u-sing different concentrations of the ethyl acetate extract of the whole herb of Prunella in breast cancer MDA-MB-231 cells for 24 h.The proliferation of MDA-MB-231 cells and the effects on the cell cycle and apoptosis of MDA-MB-231 cells were detected by using the CCK-8 assay,the cell cycle flow and the apoptotic cell flow.Western blot was used to detect the effect of ethyl ace-tate extract of whole herb of Prunella on the expression of apoptosis-related proteins in breast cancer MDA-MB-231 cells.UPLCQ-TOF MS/MS was used to detect the chemical compositions of the ethyl acetate extract of Prunella whole herb.Results The whole herb of Pru-nella had no significant effect on the growth of nude mice(P＞0.05);it could significantly inhibit the growth of transplanted tumors in nude mice with human breast cancer(P＜0.05);the results of HE staining showed that the cells in the tissues appeared to be rela-tively sparse with the increase of the dose of Prunella and had different degrees of nuclear consolidation and deep staining of nuclei and the apoptosis of the tumor cells increased;the metastasis of tumor cells to the liv-er and lungs was inhibited,when compared with that in the model group.Compared with the model group,the low,medium and high groups of Prunella had no signif-icant effect on the liver index,while the spleen index was significantly reduced(P＜0.05);the expression of Ki-67 and CK17 was reduced.The ethyl acetate ex-tract of the whole herb of Prunella could inhibit the proliferation of MDA-MB-231 cells in breast cancer(P＜0.01);the results of flow cytometry showed that,with the increase of the concentration of the ethyl ace-tate extract of the whole herb of Prunella,the proportion of S-phase cells in the MDA-MB-231 cells significantly increased,and the proportion of G0/G1-phase cells sig-nificantly decreased,while the proportion of G2-phase cells did not change significantly(P＜0.01);Western blotting was not affected in the low,medium and high groups,and the spleen index significantly decreased(P＜0.05);the expression of Ki-67 and CK17 was re-duced;the results of Western blot showed that the eth-yl acetate extract of the whole herb of Prunella promo-ted the expression of Bax,cleaved caspase-3,cleaved caspase-9 proteins,and inhibited the expression of Bcl-2,caspase-3,caspase-9,cyclinA2,and CDK2 proteins(P＜0.05,P＜0.01).The acetic acid of the whole herb of Prunella ethyl ester extract identified a total of 51 compounds.Conclusions The whole herb of Pru-nella can inhibit the growth of breast cancer in nude mice transplanted with tumors,promote the apoptosis of tumor cells,inhibit the proliferation of breast cancer MDA-MB-231 cells,inhibit the metastasis of tumor cells to the liver and lungs,protect the liver and spleen,and reduce the expression of the value-added markers Ki-67 and CK17 in tumor tissues,and the ef-fective ingredient of the whole herb,the ethyl acetate extract,can induce apoptosis.The mechanism may be related to the down-regulation of cyclinsA2,CDK2,Bcl-2,caspase-3,caspase-9 and up-regulation of Bax,cleaved caspase-3,cleaved caspase-9 protein expres-sion.

AIM To investigate the mechanism of Shaoyao Gancao Decoction in preventing meglumine diatrizoate-induced post-ERCP pancreatitis in rats through autophagy regulation.METHODS The rats were randomized into the normal group,the model group,the low-dose and high-dose Shaoyao Gancao Decoction(1.5,3.0 g/kg),and the indomethacin suppository group.A rat model of post-ERCP pancreatitis was induced by meglumine diatrizoate injection into the pancreatic duct under continuous pressure.The rats had their pancreatic tissues stained with HE to observe the pathological alterations,inflammatory cell infiltration,hemorrhage and necrosis;their serum levels of IL-1β,IL-6,IL-8,TNF-α,AMS,and IL-10 identified by ELISA;their autophagic vacuoles in pancreatic acinar cells observed by transmission electron microscopy;their pancreatic protein expressions of Beclin1,LC3B,p62,TRAF2 and p-JNK detected by IHC and Western blot;and their pancreatic mRNA expressions of Beclin1 and TRAF2 detected by RT-qPCR.RESULTS Compared with the model group,the high-dose Shaoyao Gancao Decoction group displayed no obvious hemorrhage;improvement in edema of acinar and interstitial cells;obviously less cellular inflammatory infiltration;substantially decreased serum levels of IL-1β,IL-6,TNF-α and AMS(P＜0.05,P＜0.01);drastically reduced amount of autophagosomes in acinar cells;and down-regulated expressions of autophagy-related proteins Beclin1,LC3,p62,TRAF2 and p-JNK(P＜0.05,P＜0.01).CONCLUSION Shaoyao Gancao Decoction can prevent post-ERCP pancreatitis by ameliorating pancreatic tissue injury,decreasing serum inflammatory response level,and interfering with abnormal autophagy of pancreatic acinar cells.Its molecular mechanism may involve inhibition of TRAF2 protein expression and modulation of p-JNK activation.

Aim To study the effects of whole herb of Prunella and its active components on the malignant progression of breast cancer and its mechanism.Meth-ods Breast cancer transplantation tumor model was constructed and randomly divided into the model group,low,medium and high dose group of whole herb of Prunella(0.1,0.2,0.4 g·mL-1 by gavage)and paclitaxel(10 mg·kg-1 by intraperitoneal injection),which was administered by gavage every day,and the tumor tissues were collected after 28 days of interven-tion.The weight,tumor volume and mass of nude mice in each group were detected,HE staining was used to observe the morphology of breast cancer tumor tissues,and immunohistochemical staining was used to observe the proliferation of cell-cycle regulatory protein-67(Ki-67)and cytokeratin 17(CK17)in breast cancer tumor tissues.The cellular experiments were performed by u-sing different concentrations of the ethyl acetate extract of the whole herb of Prunella in breast cancer MDA-MB-231 cells for 24 h.The proliferation of MDA-MB-231 cells and the effects on the cell cycle and apoptosis of MDA-MB-231 cells were detected by using the CCK-8 assay,the cell cycle flow and the apoptotic cell flow.Western blot was used to detect the effect of ethyl ace-tate extract of whole herb of Prunella on the expression of apoptosis-related proteins in breast cancer MDA-MB-231 cells.UPLCQ-TOF MS/MS was used to detect the chemical compositions of the ethyl acetate extract of Prunella whole herb.Results The whole herb of Pru-nella had no significant effect on the growth of nude mice(P＞0.05);it could significantly inhibit the growth of transplanted tumors in nude mice with human breast cancer(P＜0.05);the results of HE staining showed that the cells in the tissues appeared to be rela-tively sparse with the increase of the dose of Prunella and had different degrees of nuclear consolidation and deep staining of nuclei and the apoptosis of the tumor cells increased;the metastasis of tumor cells to the liv-er and lungs was inhibited,when compared with that in the model group.Compared with the model group,the low,medium and high groups of Prunella had no signif-icant effect on the liver index,while the spleen index was significantly reduced(P＜0.05);the expression of Ki-67 and CK17 was reduced.The ethyl acetate ex-tract of the whole herb of Prunella could inhibit the proliferation of MDA-MB-231 cells in breast cancer(P＜0.01);the results of flow cytometry showed that,with the increase of the concentration of the ethyl ace-tate extract of the whole herb of Prunella,the proportion of S-phase cells in the MDA-MB-231 cells significantly increased,and the proportion of G0/G1-phase cells sig-nificantly decreased,while the proportion of G2-phase cells did not change significantly(P＜0.01);Western blotting was not affected in the low,medium and high groups,and the spleen index significantly decreased(P＜0.05);the expression of Ki-67 and CK17 was re-duced;the results of Western blot showed that the eth-yl acetate extract of the whole herb of Prunella promo-ted the expression of Bax,cleaved caspase-3,cleaved caspase-9 proteins,and inhibited the expression of Bcl-2,caspase-3,caspase-9,cyclinA2,and CDK2 proteins(P＜0.05,P＜0.01).The acetic acid of the whole herb of Prunella ethyl ester extract identified a total of 51 compounds.Conclusions The whole herb of Pru-nella can inhibit the growth of breast cancer in nude mice transplanted with tumors,promote the apoptosis of tumor cells,inhibit the proliferation of breast cancer MDA-MB-231 cells,inhibit the metastasis of tumor cells to the liver and lungs,protect the liver and spleen,and reduce the expression of the value-added markers Ki-67 and CK17 in tumor tissues,and the ef-fective ingredient of the whole herb,the ethyl acetate extract,can induce apoptosis.The mechanism may be related to the down-regulation of cyclinsA2,CDK2,Bcl-2,caspase-3,caspase-9 and up-regulation of Bax,cleaved caspase-3,cleaved caspase-9 protein expres-sion.

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 clinical characteristics and prognosis of pneumococcal meningitis(PM),and drug sensitivity of Streptococcus pneumoniae(SP)isolates in Chinese children.Methods A retrospective analysis was conducted on clinical information,laboratory data,and microbiological data of 160 hospitalized children under 15 years old with PM from January 2019 to December 2020 in 33 tertiary hospitals across the country.Results Among the 160 children with PM,there were 103 males and 57 females.The age ranged from 15 days to 15 years,with 109 cases(68.1% )aged 3 months to under 3 years.SP strains were isolated from 95 cases(59.4% )in cerebrospinal fluid cultures and from 57 cases(35.6% )in blood cultures.The positive rates of SP detection by cerebrospinal fluid metagenomic next-generation sequencing and cerebrospinal fluid SP antigen testing were 40% (35/87)and 27% (21/78),respectively.Fifty-five cases(34.4% )had one or more risk factors for purulent meningitis,113 cases(70.6% )had one or more extra-cranial infectious foci,and 18 cases(11.3% )had underlying diseases.The most common clinical symptoms were fever(147 cases,91.9% ),followed by lethargy(98 cases,61.3% )and vomiting(61 cases,38.1% ).Sixty-nine cases(43.1% )experienced intracranial complications during hospitalization,with subdural effusion and/or empyema being the most common complication[43 cases(26.9% )],followed by hydrocephalus in 24 cases(15.0% ),brain abscess in 23 cases(14.4% ),and cerebral hemorrhage in 8 cases(5.0% ).Subdural effusion and/or empyema and hydrocephalus mainly occurred in children under 1 year old,with rates of 91% (39/43)and 83% (20/24),respectively.SP strains exhibited complete sensitivity to vancomycin(100% ,75/75),linezolid(100% ,56/56),and meropenem(100% ,6/6).High sensitivity rates were also observed for levofloxacin(81% ,22/27),moxifloxacin(82% ,14/17),rifampicin(96% ,25/26),and chloramphenicol(91% ,21/23).However,low sensitivity rates were found for penicillin(16% ,11/68)and clindamycin(6% ,1/17),and SP strains were completely resistant to erythromycin(100% ,31/31).The rates of discharge with cure and improvement were 22.5% (36/160)and 66.2% (106/160),respectively,while 18 cases(11.3% )had adverse outcomes.Conclusions Pediatric PM is more common in children aged 3 months to under 3 years.Intracranial complications are more frequently observed in children under 1 year old.Fever is the most common clinical manifestation of PM,and subdural effusion/emphysema and hydrocephalus are the most frequent complications.Non-culture detection methods for cerebrospinal fluid can improve pathogen detection rates.Adverse outcomes can be noted in more than 10% of PM cases.SP strains are high sensitivity to vancomycin,linezolid,meropenem,levofloxacin,moxifloxacin,rifampicin,and chloramphenicol.[Chinese Journal of Contemporary Pediatrics,2024,26(2):131-138]