The interstitial protein UL14 of pseudorabies virus(PRV)constitutes a crucial compo-nent for viral replication and virulence invasion.It can interact with other viral proteins to complete the infection of the host,rendering it one of the potentially important antiviral targets.In this stud-y,the PRV DX strain isolated in the laboratory was used as the parental strain.The recombinant UL14 protein was expressed and purified through the prokaryotic expression system.BALB/c mice were immunized with this protein as an antigen.After three rounds of subcloning screening,three hybridoma cell lines against the UL14 protein were obtained,named 1B4,1B5,and 1F2,respective-ly.The immunoreactivity of the antibodies was detected by immunofluorescence assay(IFA).The results showed that the antibody titers of the 1B4 and 1B5 strains were not lower than 1∶3 200,and that of the 1F2 strain was not lower than 1∶2 000.The results of western blot(WB)detection showed that the antibody titers of the three strains were all not lower than 1∶8 000.The results of indirect ELISA detection showed that the antibody titers in ascites were all not lower than 112 800.The results of the identification of the antigenic epitopes of the UL14 protein showed that the antigenic epitope recognized by 1B4 was 1 MFASDRRERRVRLAEAFQRE20,and the antigenic epitopes recognized by 1B5 and 1F2 were speculated to be 33GRADKKNPEFVRAFMAAKQAR53.After PRV infected susceptible cells,the temporal expression of the UL14 protein was detected by IF A using the prepared monoclonal antibodies.It was found that the temporal expression of UL14 was similar to that of gC,and the result of RT-qPCR of UL14 gene was consistent with IF A,indi-cating that UL14 might be a late gene during the PRV replication.The subcellular localization of UL14 after virus infection was detected by IFA,and it was found that the UL14 protein could be transferred from the cytoplasm to the nucleus and the expression of UL14 protein increased with the extension of infection time.This study provides a good research tool for further investigating on the function of the PRV UL14 protein and might contribute to the further study of the PRV replication mechanism mediated by the PRV UL14 protein.

The interstitial protein UL14 of pseudorabies virus(PRV)constitutes a crucial compo-nent for viral replication and virulence invasion.It can interact with other viral proteins to complete the infection of the host,rendering it one of the potentially important antiviral targets.In this stud-y,the PRV DX strain isolated in the laboratory was used as the parental strain.The recombinant UL14 protein was expressed and purified through the prokaryotic expression system.BALB/c mice were immunized with this protein as an antigen.After three rounds of subcloning screening,three hybridoma cell lines against the UL14 protein were obtained,named 1B4,1B5,and 1F2,respective-ly.The immunoreactivity of the antibodies was detected by immunofluorescence assay(IFA).The results showed that the antibody titers of the 1B4 and 1B5 strains were not lower than 1∶3 200,and that of the 1F2 strain was not lower than 1∶2 000.The results of western blot(WB)detection showed that the antibody titers of the three strains were all not lower than 1∶8 000.The results of indirect ELISA detection showed that the antibody titers in ascites were all not lower than 112 800.The results of the identification of the antigenic epitopes of the UL14 protein showed that the antigenic epitope recognized by 1B4 was 1 MFASDRRERRVRLAEAFQRE20,and the antigenic epitopes recognized by 1B5 and 1F2 were speculated to be 33GRADKKNPEFVRAFMAAKQAR53.After PRV infected susceptible cells,the temporal expression of the UL14 protein was detected by IF A using the prepared monoclonal antibodies.It was found that the temporal expression of UL14 was similar to that of gC,and the result of RT-qPCR of UL14 gene was consistent with IF A,indi-cating that UL14 might be a late gene during the PRV replication.The subcellular localization of UL14 after virus infection was detected by IFA,and it was found that the UL14 protein could be transferred from the cytoplasm to the nucleus and the expression of UL14 protein increased with the extension of infection time.This study provides a good research tool for further investigating on the function of the PRV UL14 protein and might contribute to the further study of the PRV replication mechanism mediated by the PRV UL14 protein.

OBJECTIVE To understand the prevalence of non-bacterial pathogens infections among the children who were hospitalized in pediatrics department due to acute respiratory tract infections(ARTI)from 2020 to 2024.METHODS The children who were hospitalized in pediatrics department of Lishui People's Hospital due to ARTI from Mar.2020 to Feb.2024 were enrolled in the study,the results of tests of 13 items of respiratory tract patho-gens for the children were retrospectively analyzed.The positive isolation rates of respiratory tract pathogens and epidemiological characteristics were observed and compared among the children according to the gender,age and season.RESULTS A total of 12,291 children were enrolled in the study,6508 of whom were tested positive for re-spiratory tract pathogens,and the total positive isolation rate was 52.95％.Rhinovirus(HRV),Mycoplasma pneumoniae and respiratory syncytial virus(RSV)ranked the top 3 pathogens tested positive.The total positive rate of isolation of the male children was 54.43％(3707/6810),higher than 51.10％(2801/5481)of the female children(x2=13.524,P＜0.001);the positive rates of HRV and RSV of the male children were higher than those of the female children(P＜0.05),while the positive rate of MP of the female children was higher than that of the male children(P＜0.05).There was significant difference in the total positive rate of isolation of respirato-ry tract pathogens among the age groups(x2=59.924,P＜0.001).The positive rates of RSV,HRV and MP of the infant group,the toddler group,the preschool age group and the school age group were relatively high.The total positive rate of isolation of the respiratory tract pathogens showed an upward trend with the change of sea-sons(x2=145.887,P＜0.001);there was significant difference in the prevalence of pathogens among the seasons(P＜0.05).CONCLUSIONS HRV,MP and RSV are dominant among the respiratory tract non-bacterial patho-gens isolated from the children who are hospitalized in Lishui People's Hospital from 2020 to 2024.There are sig-nificant differences in the positive rates of the respiratory tract pathogens among the genders,age,and seasons.

OBJECTIVE To evaluate the performance of commonly used molecular biological identification methods in differential identification of Citrobacter freundii complex(Cfc)and find out the differences in the mass spectra among the prevalent species of Cfc so as to facilitate the rapid and accurate identification.METHODS Totally 150 clinical strains were isolated from The First Medical Center of Chinese PLA General Hospital from 2015 to 2020 and were identified as Cfc by means of matrix assisted laser desorption ionization-time-of-flight mass spectrometry(MALDI-TOF MS),the genome data were obtained by the next generation sequencing and were compared with reference genome of Citrobacter species from National Center of Biotechnology Information(NCBI)based on aver-age nucleotide identity(ANI),phylogenetic analyses of rec N gene,concatenated alignments of four housekeeping genes(fusA,pyrG,leuS and rpoB as well as gyrB,rplB,fusA and leuS),core genome single nucleotide poly-morphism(coreSNP)and whole genome(WG),digital DNA-DNA hybridization(dDDH)analysis,ribosomal multilocus sequence typing(rMLST)and typical(strain)genome service(TYGS)analysis.ANI was set as the'gold standard' for the identification of the Cfc strains,and the capabilities of the various methods were evaluated.A certain number of strains of Citrobacter freundii(Cfr),Citrobacter braakii(Cbr)and Citrobacter portucalen-sis(Cpo)were randomly selected to obtain and compare the mass spectra so as to find out the differences in the mass spectra.RESULTS The identification capability of the Cfc strains is limited based on the available database for MALDI-TOF MS,Cpo was prone to be misidentified as Cfr or Cbr.The mass spectrum between Cpo and Cfr ranged between 2600 and 2660 m/z,the mass spectrum between Cpo and Cbr ranged between 5200 and 5300 m/z,showing significant difference.The concatenated alignments of four housekeeping genes gyrB,rplB,fusA and leuS,CoreSNP and WG phylogenetic analysis had high accurate rate of identification.CONCLUSIONS The meth-ods have certain limitations in identification of Cfc.The joint application of ANI with the methods such as WG phylogenetic analysis,update of mass spectrum database and timely establishment of database can achieve the ac-curate identification of the species of Cfc.

OBJECTIVE To understand the prevalence of non-bacterial pathogens infections among the children who were hospitalized in pediatrics department due to acute respiratory tract infections(ARTI)from 2020 to 2024.METHODS The children who were hospitalized in pediatrics department of Lishui People's Hospital due to ARTI from Mar.2020 to Feb.2024 were enrolled in the study,the results of tests of 13 items of respiratory tract patho-gens for the children were retrospectively analyzed.The positive isolation rates of respiratory tract pathogens and epidemiological characteristics were observed and compared among the children according to the gender,age and season.RESULTS A total of 12,291 children were enrolled in the study,6508 of whom were tested positive for re-spiratory tract pathogens,and the total positive isolation rate was 52.95％.Rhinovirus(HRV),Mycoplasma pneumoniae and respiratory syncytial virus(RSV)ranked the top 3 pathogens tested positive.The total positive rate of isolation of the male children was 54.43％(3707/6810),higher than 51.10％(2801/5481)of the female children(x2=13.524,P＜0.001);the positive rates of HRV and RSV of the male children were higher than those of the female children(P＜0.05),while the positive rate of MP of the female children was higher than that of the male children(P＜0.05).There was significant difference in the total positive rate of isolation of respirato-ry tract pathogens among the age groups(x2=59.924,P＜0.001).The positive rates of RSV,HRV and MP of the infant group,the toddler group,the preschool age group and the school age group were relatively high.The total positive rate of isolation of the respiratory tract pathogens showed an upward trend with the change of sea-sons(x2=145.887,P＜0.001);there was significant difference in the prevalence of pathogens among the seasons(P＜0.05).CONCLUSIONS HRV,MP and RSV are dominant among the respiratory tract non-bacterial patho-gens isolated from the children who are hospitalized in Lishui People's Hospital from 2020 to 2024.There are sig-nificant differences in the positive rates of the respiratory tract pathogens among the genders,age,and seasons.

OBJECTIVE To evaluate the performance of commonly used molecular biological identification methods in differential identification of Citrobacter freundii complex(Cfc)and find out the differences in the mass spectra among the prevalent species of Cfc so as to facilitate the rapid and accurate identification.METHODS Totally 150 clinical strains were isolated from The First Medical Center of Chinese PLA General Hospital from 2015 to 2020 and were identified as Cfc by means of matrix assisted laser desorption ionization-time-of-flight mass spectrometry(MALDI-TOF MS),the genome data were obtained by the next generation sequencing and were compared with reference genome of Citrobacter species from National Center of Biotechnology Information(NCBI)based on aver-age nucleotide identity(ANI),phylogenetic analyses of rec N gene,concatenated alignments of four housekeeping genes(fusA,pyrG,leuS and rpoB as well as gyrB,rplB,fusA and leuS),core genome single nucleotide poly-morphism(coreSNP)and whole genome(WG),digital DNA-DNA hybridization(dDDH)analysis,ribosomal multilocus sequence typing(rMLST)and typical(strain)genome service(TYGS)analysis.ANI was set as the'gold standard' for the identification of the Cfc strains,and the capabilities of the various methods were evaluated.A certain number of strains of Citrobacter freundii(Cfr),Citrobacter braakii(Cbr)and Citrobacter portucalen-sis(Cpo)were randomly selected to obtain and compare the mass spectra so as to find out the differences in the mass spectra.RESULTS The identification capability of the Cfc strains is limited based on the available database for MALDI-TOF MS,Cpo was prone to be misidentified as Cfr or Cbr.The mass spectrum between Cpo and Cfr ranged between 2600 and 2660 m/z,the mass spectrum between Cpo and Cbr ranged between 5200 and 5300 m/z,showing significant difference.The concatenated alignments of four housekeeping genes gyrB,rplB,fusA and leuS,CoreSNP and WG phylogenetic analysis had high accurate rate of identification.CONCLUSIONS The meth-ods have certain limitations in identification of Cfc.The joint application of ANI with the methods such as WG phylogenetic analysis,update of mass spectrum database and timely establishment of database can achieve the ac-curate identification of the species of Cfc.

As the backbone force of China's social and economic construction, the health status of workers is closely related to the nation's productivity and social development. Currently, cancers have become one of the major diseases threatening the health of workers. However, there are still many shortcomings in the cancer screening services for the workers. To standardize cancer screening services for workers, ensure the quality of screening services, and improve the overall screening effectiveness, 19 institutions, including Peking Union Medical College Hospital of the Chinese Academy of Medical Sciences, have jointly formulated the Group Standard "Specification for service of cancer screening for workers (T/CHAA 023-2023)". This standard follows the principles of "legality, scientific rigor, advancement, and feasibility" and combines the frontier scientific advances in cancer screening. It clarifies the relevant requirements for service principles, service design, service delivery, service management, service evaluation, and improving worker cancer screening. Implementing this group standard will help connect the common screening needs of workers, employers, and cancer screening service providers, standardize the screening process, improve screening quality, and ultimately increase the early diagnosis rate and survival rate of cancer patients. Consequently, this group standard will help safeguard workers' health rights and interests, ensure the labor force resources, promote the comprehensive coordinated and sustainable development of society, and contribute to realizing the "Healthy China 2030" strategic policy.

This article reports the pharmacotherapy process of a clinical pharmacist participating in the treatment of acute liver injury caused by nintedanib in a patient with lung cancer complicated with interstitial pneumonia.Clinical pharmacist analyzed the rapid increase of liver enzymes in patients by excluding their past drug use history,stopping suspicious drugs and searching domestic and foreign literature,and determined that the association with nidanib was very likely according to RUCAM scale.The clinical pharmacies suggesteded stopping nintedanib and taking liver protection treatment actively.The clinician adopted the suggestion,and finally the patient's liver function improved after 20 days of treatment.Clinical pharmacists demonstrate their value by analyzing the causes and characteristics of liver injury in patients,assisting physicians in the timely identification and management of adverse reactions,and participating in clinical practice.This article can provide a reference for the diagnosis and treatment of similar cases of acute liver injury.

Objective To provide alternative delivery methods of novel oral antitumor drugs for patients with dysphagia.Methods The retrieval range was determined according to the"Guidelines for Clinical Application of Novel Antitumor Drugs(2022 edition)".By consulting the instructions and searching databases such as PubMed,Micromedex,UpToDate,etc.,the preparation,stability,storage,and related clinical information of temporary liquid drug formulations were obtained.Results Seventy novel oral anticancer drugs were included in the literature search.Thirty-three drugs had relevant literature or data supporting alternative administration methods,only eight had information on alternative administration in the instructions,and the level of evidence for other drugs varied.Conclusion The evidence levels were low for most temporary liquid formulations,and medical teams should fully consider the advantages and disadvantages of using these products outside the instructions and use them with caution.

Objective : The purpose of this study was to investigate the effects of plasma exosome⁃derived miR⁃29b⁃3p in myocardial ischemia⁃reperfusion injury ( MIRI) rats on hypoxia/reoxygenation ( H/R) cardiomyocyte after sevoflurane (SEV) postconditioning through targeting IGF1 . Methods : The GEO database was used to screen differentially expressed miRNAs in MIRI , and cardiomyocytes were treated with H/R to construct a MIRI cell model. The expression of miR⁃29b⁃3p and IGF1 in the MIRI cell model post⁃treated with SEV was intervened , and then the survival rate of cardiomyocytes was detected by MTT , apoptosis was detected by flow cytometry , and inflammatory factors (IL⁃1β and TNF⁃α ) in cardiomyocytes in each group were detected by ELISA. Results : Compared with Normal group , the expression of miR⁃29b⁃3p in plasma exosomes of MIRI rats was enhanced (P < 0. 05) , and the target⁃binding relationship between miR⁃29b⁃3p and IGF1 was confirmed ( P < 0. 05) . After SEV post⁃treatment , the expression of miR⁃29b⁃3p in H/R⁃stimulated cardiomyocytes decreased , while the expression of IGF1 increased (both P < 0. 05) . Overexpression of miR⁃29b⁃3p in plasma exosomes could significantly inhibit the survival rate of H/R cells after SEV treatment , aggravate apoptosis and inflammatory response , while knockdown of miR⁃29b⁃3p showed a opposite effects (all P < 0. 05) . The rescue experimental data showed that overexpression of IGF1 could partially reverse the effects of overexpression of miR⁃29b⁃3p on H/R cell injury after SEV treatment ( all P <0. 05) . Conclusion Plasma exosome⁃derived miR⁃29b⁃3p promotes H/R cardiomyocyte injury after SEV treatment by targeting IGF1 .