To develop a novel immunocastration vaccine for animals,researchers designed and syn-thesized the recombinant plasmid pET-30a-SF which could express the recombinant protein SF.This protein was then conjugated in vitro with the synthetic peptide STGP to prepare the SF-STGP nanoparticle vaccine,and its immunocastration effect on mice was studied.The Spy Catcher and ferritin amino acid sequences were connected via GGGGS,and after codon optimization for E.coli,the recombinant plasmid pET-30a-SF was constructed and transformed into E.coli for in-duced expression.The recombinant protein SF was purified using Ni-column affinity chromatogra-phy and characterized.The peptide STGP,composed of Spy Tag,GnRH,and PADRE connected by GGGS,was conjugated with the recombinant protein SF in vitro.The self-assembled nanoparticles were observed using transmission electron microscopy(TEM)and dynamic light scattering(DLS).The prepared SF-STGP nanoparticles were mixed with MONTANIDE ISA 206 VG at a 1∶1 ratio to form the vaccine,which was then subcutaneously injected into male BALB/c mice for immunocastration evaluation.The recombinant protein SF showed the highest soluble expression when induced at 18 ℃ with 0.25 mmol/L IPTG for 14 h,and the maximum conjugation efficiency with STGP was achieved at a 1∶8 molar ratio.TEM and DLS analyses revealed that both the re-combinant protein SF and SF-STGP could self-assemble into nanoparticles with average diameters of 16.2 nm and 17.8 nm,respectively.Mouse immunization results demonstrated that the SF-STGP nanoparticle vaccine generated specific GnRH antibodies after the first immunization,with the spe-cific antibody D45o reaching its peak at the 10 th week.The SF-STGP+ISA 206 immunization group showed a peak D450 value of 2.8,and the specific antibody levels in all immunization groups were significantly higher than those in the control group(P＜0.05).Additionally,the SF-STGP nanoparticle vaccine effectively reduced serum testosterone levels in mice,with the testosterone concentration in the immunization groups being significantly lower than that in the control group(P＜0.05).Compared to the control group,the immunization group exhibits testicular atrophy.The constructed SF-STGP nanoparticle vaccine proves to be a highly effective immunogen,capable of inducing testicular atrophy and reducing gonadal hormone concentrations,demonstrating excellent castration effects.This study provides new insights into immunocastration vaccines for mammals.

OBJECTIVE To explore the distribution and drug resistance trends of the main pathogens causing blood-stream infections(BSI)in neonatal intensive care unit(NICU)of Henan Province from 2014 to 2023 so as to pro-vide bases for prevention and control of hospital-associated infections in the neonates and reasonable use of antibi-otics in the whole province.METHODS The data regarding to the pathogens causing BSI in the NICU neonates and drug resistance were retrospectively collected from Jan.2014 to Dec.2023,and the statistical analysis was per-formed by SPSS 26.0 and WHONET 5.6 software.RESULTS Totally 27,984 strains of pathogens were collected from 2014 to 2023,13,547(48.41％)of which were gram-negative bacteria,and 14,437(51.59％)were gram-pos-itive bacteria.Klebsiella pneumoniae(4221 strains,15.08％),Escherichia coli(3735 strains,13.35％),Acine-tobacter baumannii(1288 strains,4.60％),Enterobacter cloacae(847 strains,3.12％)and Pseudomonas aerugi-nosa(655 strain,2.34％)were the major species of gram-negative bacteria;Staphylococcus aureus(4545 strains,16.24％),Staphylococcus epidermidis(3306 strains,11.81％),Staphylococcus haemolyticus(2048 strains,7.32％),Staphylococcus hominis(1085 strains,3.88％)and Enterococcus faecalis(946 strains,3.38％)were the predominant species of gram-positive bacteria.The drug resistant analysis indicated that the drug resistance rate of the K.pneumoniae strains to imipenem showed an upward trend during the past six years,peaking at 44.23％,and it began to decline in 2021;though the drug resistance rate of the E.coli strains to imipenem showed some fluctuations,it gener-ally presented a downward trend,peaking at 7.00％.The drug resistance rates of the K.pneumoniae strains to the third generation of cephalosporins and carbapenems were higher than those of the E.coli,and there was significant differ-ence in the antimicrobial prevalence trend between the two species of Enterobacter during the ten years(P＜0.05).CONCLUSIONS K.pneumoniae and coagulase-negative Staphylococcus are dominant among the pathogens causing BSI in the ICU neonates of the whole province,and the isolation rates of drug-resistant strains are high.It is grossly necessary for the reasonable clinical use of antibiotics to carry out the bacterial drug resistance surveillance.

Objective:To analyze the clinical phenotypes and genetic characteristics of pediatric patients with neurofibromatosis type 1 (NF1).Methods:A cross-sectional study was adopted. Clinical and imaging data of 25 pediatric patients diagnosed as having NF1 in Department of Pediatric Neurology, Linyi People's Hospital Affiliated to Shandong Second Medical University from January 2024 to July 2025 were collected. Whole exome sequencing and Sanger sequencing were used to conduct genetic testing on the pediatric patients and his/her parents. Protein 3D modeling of the domestic and foreign unreported variations was conducted using SWISS-MODEL software.Results:Among the 25 pediatric patients with NF1, 14 were male (56%) and 11 were female (44%), with age ranging from 8 months to 18 years. All pediatric patients exhibited café-au-lait macules, and 7 (28%) presented with plexiform neurofibromas. Genetic test identified 4 types of NF1 variants: nonsense variant ( n=11, 44%), frameshift variant ( n=9, 36%), missense variant ( n=3, 12%), and splice-site variant ( n=2, 8%). Importantly, 5 novel NF1 variants were discovered, including c.3455T>A, c.3709dupG, c.2665_2684del, c.7092_7095delinsTA, and c.3260del. Three pediatric patients inherited NF1 variant from their parents, while the remaining 22 harbored de novo mutation. Conclusion:NF1 exhibits a broad clinical spectrum, primarily affecting the skin and nervous system; this study identifies 5 previously unreported variants, expanding the genetic profile of NF1.

Objective To investigate the mechanism of action and potential biomarkers of Jiangzhi Qingshen capsules in the treatment of dyslipidaemia based on clinical lipid metabolomics.Methods 30 patients with dyslipidaemia from Zhengzhou Hospital of Chinese Medicine were collected as the test group,and their lipid levels before and after taking Jiang Zhi Qing Shen capsule for 12 weeks were compared.Another 30 healthy patients were enrolled in the physical examination department of Zhengzhou Hospital of Chinese Medicine,and metabolomics investigation was carried out on plasma samples of the test group before and after the treatment as well as those of the healthy patients by LC-MS/MS technology.The differences between the groups were compared by multivariate statistical analysis,and the potential biomarkers were identified by HMDB and lipidblast databases,so as to clarify the possible pathways and targets for the treatment of mild dyslipidaemia by Jiangzhi Qingshen capsules.Results Jiangzhi Qingshen capsules could improve patients' blood lipid level,BMI and abdominal circumference significantly(P<0.05).And metabolomics results showed that 29 lipid metabolites in the plasma of the treated patients were dialled back,which involved cholesterol metabolism,fat digestion and absorption,glycerol-phospholipid metabolism and other biological processes.Conclusion The efficacy of Jiangzhi Qingshen capsules in treating dyslipidaemia was confirmed,and its mechanism of action might be related to the regulation of lipid metabolism.Metabolites such as acylcarnitine(Acar),phosphatidylethanolamine(PE)and phosphatidylcholine(PC)were expected to be used as the biomarkers of dyslipidaemia,so as to provide objective scientific basis for the lipid-lowering capsule,and to facilitate the promotion of its application.

Objective To investigate the clinical efficacy of the traditional Chinese medicine(TCM)throat opening and brightening method in treating unilateral vocal cord paralysis(UVCP).Methods Sixty patients with UVCP were prospectively collected and randomly assigned to two groups:the Chinese herbal medicine group(trea-ted with Buyang Huanwu Decoction,n=30)and the throat opening and brightening method group(treated with TCM throat opening and brightening method,n=30).The clinical studies that had utilized injection laryngoplasty for the treatment of UVCP(historical control group).Evaluation indicators included the voice handicap index-10(VHI-10),GRBAS-G,objective acoustic measurements of voice(vocal intensity,F0,shimmer,jitter,HNR),and aerodynamic measurements(maximum phonation time,MPT).Results Before treatment,no significant differences were observed between the two groups in all the evaluation indicators(P＞0.05).Post-treatment,the throat open-ing and brightening method group demonstrated significant improvements in VHI-10,GRBAS-G,shimmer,jitter,HNR,and MPT compared to pre-treatment values(P＜0.01),and these improvements were superior to those in the Chinese herbal medicine group.Pre-treatment VHI-10,GRBAS-G,and shimmer scores in the throat opening and brightening method group were significantly higher than those in the historical literature group(P＜0.01).Af-ter treatment,no significant differences were noted in any assessed parameters between the two groups(P＞0.05).Conclusion The TCM throat opening and brightening method significantly enhances phonatory function and quality of life in patients with UVCP,showing comparable efficacy to injection laryngoplasty.

Objective To observe the effects of Jianpi Huatan Prescription on hepatic cholesterol synthesis in subclinical hypothyroidism(SCH)mice;To discuss its mechanism based on cAMP/CREB/HMGCR signaling pathway.Methods Totally 80 C57BL/6 male mice were randomly divided into control group(10 mice)and modeling group(70 mice),and the modeling group was given methimazole 0.08 mg/(kg·d)in drinking water for 16 weeks to establish a SCH mdoel.The model mice were randomly divided into model group,euthyrox group and Jianpi Huatan Prescription high-,medium-and low-dosage groups,with 10 mice in each group,and were given corresponding drugs for gavage for 6 weeks.HE staining and Oil red O staining were used to observe the morphology and lipid deposition of liver tissue,ELISA was used to detect serum contents of thyroid stimulating hormone(TSH),triiodothyronine(T3),thyroid hormone(T4),total cholesterol(TC),triglycerides(TG),and TC,cyclic adenosine monophosphate(cAMP)in liver tissue,Western blot was used to detect the expressions of protein kinase A(PKA),cyclic adenosine response element binding protein(CREB),p-CREB and 3-hydroxy-3-methylglutaryl-CoA reductase(HMGCR)in liver tissue.Results Compared with the control group,the liver tissue of mice in the model group showed fat vacuoles of different sizes and obvious lipid deposition;the contents of TSH,TC,TG in serum and TC,cAMP in liver tissue significantly increased(P＜0.05,P＜0.01);the protein expressions of PKA,p-CREB and HMGCR significantly increased(P＜0.01).Compared with the model group,lipid deposition in liver tissue and structure of liver cells was improved to varying degrees in euthyrox group and Jianpi Huatan Prescription high-and medium-dosage groups,and the contents of serum TSH,TC,TG,and liver tissue TC,cAMP decreased(P＜0.05,P＜0.01);the expressions of PKA,p-CREB and HMGCR protein in liver tissue increased significantly(P＜0.01).Conclusion Jianpi Huatan Prescription can significantly inhibit hepatic cholesterol synthesis in SCH mice,and improve hepatic fat vacuoles and lipid deposition,and its mechanism may be to reduce TSH levels in SCH mice by regulating the cAMP/CREB/HMGCR signaling pathway.

Objective To investigate the mechanism of action and potential biomarkers of Jiangzhi Qingshen capsules in the treatment of dyslipidaemia based on clinical lipid metabolomics.Methods 30 patients with dyslipidaemia from Zhengzhou Hospital of Chinese Medicine were collected as the test group,and their lipid levels before and after taking Jiang Zhi Qing Shen capsule for 12 weeks were compared.Another 30 healthy patients were enrolled in the physical examination department of Zhengzhou Hospital of Chinese Medicine,and metabolomics investigation was carried out on plasma samples of the test group before and after the treatment as well as those of the healthy patients by LC-MS/MS technology.The differences between the groups were compared by multivariate statistical analysis,and the potential biomarkers were identified by HMDB and lipidblast databases,so as to clarify the possible pathways and targets for the treatment of mild dyslipidaemia by Jiangzhi Qingshen capsules.Results Jiangzhi Qingshen capsules could improve patients' blood lipid level,BMI and abdominal circumference significantly(P<0.05).And metabolomics results showed that 29 lipid metabolites in the plasma of the treated patients were dialled back,which involved cholesterol metabolism,fat digestion and absorption,glycerol-phospholipid metabolism and other biological processes.Conclusion The efficacy of Jiangzhi Qingshen capsules in treating dyslipidaemia was confirmed,and its mechanism of action might be related to the regulation of lipid metabolism.Metabolites such as acylcarnitine(Acar),phosphatidylethanolamine(PE)and phosphatidylcholine(PC)were expected to be used as the biomarkers of dyslipidaemia,so as to provide objective scientific basis for the lipid-lowering capsule,and to facilitate the promotion of its application.

Objective To investigate the clinical efficacy of the traditional Chinese medicine(TCM)throat opening and brightening method in treating unilateral vocal cord paralysis(UVCP).Methods Sixty patients with UVCP were prospectively collected and randomly assigned to two groups:the Chinese herbal medicine group(trea-ted with Buyang Huanwu Decoction,n=30)and the throat opening and brightening method group(treated with TCM throat opening and brightening method,n=30).The clinical studies that had utilized injection laryngoplasty for the treatment of UVCP(historical control group).Evaluation indicators included the voice handicap index-10(VHI-10),GRBAS-G,objective acoustic measurements of voice(vocal intensity,F0,shimmer,jitter,HNR),and aerodynamic measurements(maximum phonation time,MPT).Results Before treatment,no significant differences were observed between the two groups in all the evaluation indicators(P＞0.05).Post-treatment,the throat open-ing and brightening method group demonstrated significant improvements in VHI-10,GRBAS-G,shimmer,jitter,HNR,and MPT compared to pre-treatment values(P＜0.01),and these improvements were superior to those in the Chinese herbal medicine group.Pre-treatment VHI-10,GRBAS-G,and shimmer scores in the throat opening and brightening method group were significantly higher than those in the historical literature group(P＜0.01).Af-ter treatment,no significant differences were noted in any assessed parameters between the two groups(P＞0.05).Conclusion The TCM throat opening and brightening method significantly enhances phonatory function and quality of life in patients with UVCP,showing comparable efficacy to injection laryngoplasty.

To develop a novel immunocastration vaccine for animals,researchers designed and syn-thesized the recombinant plasmid pET-30a-SF which could express the recombinant protein SF.This protein was then conjugated in vitro with the synthetic peptide STGP to prepare the SF-STGP nanoparticle vaccine,and its immunocastration effect on mice was studied.The Spy Catcher and ferritin amino acid sequences were connected via GGGGS,and after codon optimization for E.coli,the recombinant plasmid pET-30a-SF was constructed and transformed into E.coli for in-duced expression.The recombinant protein SF was purified using Ni-column affinity chromatogra-phy and characterized.The peptide STGP,composed of Spy Tag,GnRH,and PADRE connected by GGGS,was conjugated with the recombinant protein SF in vitro.The self-assembled nanoparticles were observed using transmission electron microscopy(TEM)and dynamic light scattering(DLS).The prepared SF-STGP nanoparticles were mixed with MONTANIDE ISA 206 VG at a 1∶1 ratio to form the vaccine,which was then subcutaneously injected into male BALB/c mice for immunocastration evaluation.The recombinant protein SF showed the highest soluble expression when induced at 18 ℃ with 0.25 mmol/L IPTG for 14 h,and the maximum conjugation efficiency with STGP was achieved at a 1∶8 molar ratio.TEM and DLS analyses revealed that both the re-combinant protein SF and SF-STGP could self-assemble into nanoparticles with average diameters of 16.2 nm and 17.8 nm,respectively.Mouse immunization results demonstrated that the SF-STGP nanoparticle vaccine generated specific GnRH antibodies after the first immunization,with the spe-cific antibody D45o reaching its peak at the 10 th week.The SF-STGP+ISA 206 immunization group showed a peak D450 value of 2.8,and the specific antibody levels in all immunization groups were significantly higher than those in the control group(P＜0.05).Additionally,the SF-STGP nanoparticle vaccine effectively reduced serum testosterone levels in mice,with the testosterone concentration in the immunization groups being significantly lower than that in the control group(P＜0.05).Compared to the control group,the immunization group exhibits testicular atrophy.The constructed SF-STGP nanoparticle vaccine proves to be a highly effective immunogen,capable of inducing testicular atrophy and reducing gonadal hormone concentrations,demonstrating excellent castration effects.This study provides new insights into immunocastration vaccines for mammals.

OBJECTIVE To explore the distribution and drug resistance trends of the main pathogens causing blood-stream infections(BSI)in neonatal intensive care unit(NICU)of Henan Province from 2014 to 2023 so as to pro-vide bases for prevention and control of hospital-associated infections in the neonates and reasonable use of antibi-otics in the whole province.METHODS The data regarding to the pathogens causing BSI in the NICU neonates and drug resistance were retrospectively collected from Jan.2014 to Dec.2023,and the statistical analysis was per-formed by SPSS 26.0 and WHONET 5.6 software.RESULTS Totally 27,984 strains of pathogens were collected from 2014 to 2023,13,547(48.41％)of which were gram-negative bacteria,and 14,437(51.59％)were gram-pos-itive bacteria.Klebsiella pneumoniae(4221 strains,15.08％),Escherichia coli(3735 strains,13.35％),Acine-tobacter baumannii(1288 strains,4.60％),Enterobacter cloacae(847 strains,3.12％)and Pseudomonas aerugi-nosa(655 strain,2.34％)were the major species of gram-negative bacteria;Staphylococcus aureus(4545 strains,16.24％),Staphylococcus epidermidis(3306 strains,11.81％),Staphylococcus haemolyticus(2048 strains,7.32％),Staphylococcus hominis(1085 strains,3.88％)and Enterococcus faecalis(946 strains,3.38％)were the predominant species of gram-positive bacteria.The drug resistant analysis indicated that the drug resistance rate of the K.pneumoniae strains to imipenem showed an upward trend during the past six years,peaking at 44.23％,and it began to decline in 2021;though the drug resistance rate of the E.coli strains to imipenem showed some fluctuations,it gener-ally presented a downward trend,peaking at 7.00％.The drug resistance rates of the K.pneumoniae strains to the third generation of cephalosporins and carbapenems were higher than those of the E.coli,and there was significant differ-ence in the antimicrobial prevalence trend between the two species of Enterobacter during the ten years(P＜0.05).CONCLUSIONS K.pneumoniae and coagulase-negative Staphylococcus are dominant among the pathogens causing BSI in the ICU neonates of the whole province,and the isolation rates of drug-resistant strains are high.It is grossly necessary for the reasonable clinical use of antibiotics to carry out the bacterial drug resistance surveillance.