Objective:A camelid natural nanobody library was screened to target S100 calcium-binding protein B (S100B) for obtaining high-affinity and specific nanobodies (Nbs), which provided a molecular basis for the early diagnosis and prognostic treatment of malignant melanoma.Methods:In this study, affinity panning was employed to isolate S100B-specific nanobodies with unique complementarity-determining region 3 (CDR3) sequences from a camelid natural nanobody library. The selected Nbs were expressed in a prokaryotic system and purified via Ni-NTA affinity chromatography. The affinity, specificity, and diagnostic potential of the Nbs were evaluated using enzyme-linked immunosorbent assay (ELISA), western blot, and bio-layer interferometry (BLI).Results:A camelid natural anti-S100B nanobody library with a capacity of 1.91×10 8 CFU was successfully constructed. Affinity panning yielded 30 S100B-specific Nbs, among which Nb107, Nb122, Nb212, and Nb324 with distinct CDR3 sequences were selected for expression. Following Ni-NTA purification, all four anti-S100B Nbs exhibited high purity. Western blot analysis confirmed their ability to recognize recombinant S100B. ELISA and BLI analyses revealed that Nb212 demonstrated high affinity (1.96×10 -11 mol). Additionally, Nb107, Nb122, and Nb212 exhibited broad-spectrum recognition capabilities, binding to various tumor cell lines (Hepa1-6, GL261, 4T1, CT26) as well as murine/human melanoma cells. These Nbs also effectively bound to native murine/human antigens in serum samples from melanoma (A375, B16F10) mouse models. Conclusions:Specific anti-S100B Nbs are successfully screened and expressed, demonstrating not only recognition of native conformational antigens but also broad-spectrum binding and high affinity. These findings highlight their significant potential for developing early diagnostic assays and broad-spectrum targeted vaccines or therapeutics against diverse tumor cells.

Objective To analyze the correlation of cholecystectomy and changes in intestinal microbiota composition and function by observing functional characteristics of differential microbial communities.Methods A cross-sectional study was conducted on the patients(PC group,n=73)undergoing cholecystectomy in our hospital from 2020 to 2021.Another 56 healthy age-and gender-matched individuals(HC group)without a history of cholecystectomy were subjected and served as the control group.Fecal specimens were collected from the 2 groups.16S rRNA sequencing analysis was performed to examine the changes in composition and function of intestinal microbiota.Results There were no statistical differences between the 2 groups in baseline indicators,such as gender,age,BMI,smoking and drinking history,blood pressure,heart rate,and comorbidities,but significant difference was observed in total bilirubin(TBIL)between them(P<0.01).Alpha diversity analysis showed no significant difference in Chao1,Shannon,and Simpson indices between the 2 groups.Beta diversity analysis using the Bray-Curtis distance algorithm revealed a significant difference between the 2 groups at the class and genus levels(P<0.05).The analysis of microbiota relative abundance using LEFSE showed that Enterobacteriaceae,Lactobacillales,Citrobacter,Megasphaera,Lactobacillus,Enterococcus,Akkermansia,Streptococcus,Klebsiella,and Ruminococcus_gnavus were up-regulated in the PC group,and Lachnospiraceae,Sutterellaceae,Lachnospirales,Lachnospira,and Sutterella were down-regulated.Kyoto Encyclopedia of Genes and Genomes(KEGG)functional prediction analysis indicated that significant differences were seen between the 2 groups in metabolic pathways,including ascorbic acid(vitamin C)metabolism and aldonic acid metabolism(P<0.05),tricarboxylic acid cycle(TCA cycle)(P<0.05),glutathione metabolism(P<0.05),glutamic acid metabolism(P<0.05),secondary bile acid metabolism(P<0.05),and pentose phosphate pathway(P<0.01).Conclusion Cholecystectomy is closely associated with the structural alterations in the composition of intestinal microbiota.Variations in microbiota composition and function may induce perturbations in TCA cycle and glutathione metabolism,glutamate metabolism,secondary bile acid metabolism,and pentose phosphate pathways.

Objective To explore the influencing factors of early neurological deterioration(END)in patients with acute anterior circulation large-vessel occlusive mild stroke who were treated with medications alone within 72 h after onset.Methods Retrospective consecutive data were collected of patients with acute large-vessel occlusive mild stroke who presented to the Advanced Stroke Center of Linyi People's Hospital within 24 h of onset from January 2021 to December 2022.END was defined as an increase of ≥ 4 points in the National Institutes of Health stroke scale(N1HSS)score within 72 h after onset compared to the admission score.Patients were divided into the neurological deterioration group and the stable condition group(NIHSS score did not increase or increased by 1-3 points within 72 h after onset compared to the admission score).Baseline and clinical data of all patients were collected,including sex,age,cerebrovascular disease risk factors(hypertension,diabetes mellitus,hyperlipidemia,coronary heart disease,atrial fibrillation,smoking,alcohol consumption,stroke history),NIHSS score at admission,time from onset to admission,systolic blood pressure at admission,diastolic blood pressure at admission,laboratory test indicators at admission(blood glucose,glycosylated hemoglobin,homocysteine,total cholesterol,triglyceride,low-density lipoprotein cholesterol,high-density lipoprotein cholesterol,neutrophils,lymphocytes and neutrophil-to-lymphocyte ratio),responsible occlusion artery(internal carotid artery,middle cerebral artery,anterior cerebral artery),affected cerebral hemisphere,collateral circulation score,and medications used within 72 h after admission(intravenous thrombolysis+dual antiplatelet therapy,tirofiban+dual antiplatelet therapy,argatroban+dual antiplatelet therapy,argatroban alone,dual antiplatelet therapy alone).Variables with statistically significant differences in univariate analysis were included in multivariate Logistic regression analysis to explore the independent influencing factors for END in patients with acute anterior circulation large-vessel occlusive mild stroke treated with medications alone.Results A total of 208 patients with acute anterior circulation large-vessel occlusive mild stroke were included,with 143 males and 65 females,aged 38-85 years,with an average age of(64±9)years.Among them,86 patients were in the neurological deterioration group and 122 in the stable condition group.(1)There were statistically significant differences between the neurological deterioration group and the stable condition group in terms of history of diabetes mellitus(39.5％[34/86]vs.17.2％[21/122]),smoking history(43.0％[37/86]vs.29.5％[36/122]),left cerebral hemisphere lesion(57.0％[49/86]vs.41.0％[50/122]),collateral circulation score(4[3,5]vs.5[4,5]),time from onset to admission(7.0[3.0,17.0]hvs.4.3[2.0,11.0]h),blood glucose at admission(7.4[5.8,10.0]mmol/L vs.6.7[5.8,7.7]mmol/L),neutrophil-to-lymphocyte ratio(3.8[2.4,5.1]vs.3.0[2.1,4.3]),dual antiplatelet therapy alone(19.8％[17/86]vs.6.6％[8/122]),and argatroban+dual antiplatelet therapy(8.1％[7/86]vs.29.5％[36/122];all P＜0.05).There were no statistically significant differences in the results of the remaining univariate analyses(all P＞0.05).(2)Multivariate Logistic regression analysis showed that diabetes mellitus(OR,2.674,95％CI 1.121-6.377,P=0.027)and left cerebral hemisphere vessel occlusion(OR,2.030,95％CI I.083-3.806,P=0.027)were independent risk factors for END in acute anterior circulation large-vessel occlusive mild stroke.Argatroban+dual antiplatelet therapy(OR,0.267,95％CI 0.116-0.613,P=0.002)and high collateral circulation score(OR,0.551,95％CI 0.368-0.824,P=0.004)were independent protective factors for END in acute anterior circulation large-vessel occlusive mild stroke.Conclusions Acute anterior circulation large-vessel occlusive mild stroke patients with diabetes mellitus or left cerebral hemisphere lesions are prone to END.The combination of argatroban and dual antiplatelet therapy and good collateral circulation can reduce the risk of END.

Objective:A camelid natural nanobody library was screened to target S100 calcium-binding protein B (S100B) for obtaining high-affinity and specific nanobodies (Nbs), which provided a molecular basis for the early diagnosis and prognostic treatment of malignant melanoma.Methods:In this study, affinity panning was employed to isolate S100B-specific nanobodies with unique complementarity-determining region 3 (CDR3) sequences from a camelid natural nanobody library. The selected Nbs were expressed in a prokaryotic system and purified via Ni-NTA affinity chromatography. The affinity, specificity, and diagnostic potential of the Nbs were evaluated using enzyme-linked immunosorbent assay (ELISA), western blot, and bio-layer interferometry (BLI).Results:A camelid natural anti-S100B nanobody library with a capacity of 1.91×10 8 CFU was successfully constructed. Affinity panning yielded 30 S100B-specific Nbs, among which Nb107, Nb122, Nb212, and Nb324 with distinct CDR3 sequences were selected for expression. Following Ni-NTA purification, all four anti-S100B Nbs exhibited high purity. Western blot analysis confirmed their ability to recognize recombinant S100B. ELISA and BLI analyses revealed that Nb212 demonstrated high affinity (1.96×10 -11 mol). Additionally, Nb107, Nb122, and Nb212 exhibited broad-spectrum recognition capabilities, binding to various tumor cell lines (Hepa1-6, GL261, 4T1, CT26) as well as murine/human melanoma cells. These Nbs also effectively bound to native murine/human antigens in serum samples from melanoma (A375, B16F10) mouse models. Conclusions:Specific anti-S100B Nbs are successfully screened and expressed, demonstrating not only recognition of native conformational antigens but also broad-spectrum binding and high affinity. These findings highlight their significant potential for developing early diagnostic assays and broad-spectrum targeted vaccines or therapeutics against diverse tumor cells.

Objective To explore the influencing factors of early neurological deterioration(END)in patients with acute anterior circulation large-vessel occlusive mild stroke who were treated with medications alone within 72 h after onset.Methods Retrospective consecutive data were collected of patients with acute large-vessel occlusive mild stroke who presented to the Advanced Stroke Center of Linyi People's Hospital within 24 h of onset from January 2021 to December 2022.END was defined as an increase of ≥ 4 points in the National Institutes of Health stroke scale(N1HSS)score within 72 h after onset compared to the admission score.Patients were divided into the neurological deterioration group and the stable condition group(NIHSS score did not increase or increased by 1-3 points within 72 h after onset compared to the admission score).Baseline and clinical data of all patients were collected,including sex,age,cerebrovascular disease risk factors(hypertension,diabetes mellitus,hyperlipidemia,coronary heart disease,atrial fibrillation,smoking,alcohol consumption,stroke history),NIHSS score at admission,time from onset to admission,systolic blood pressure at admission,diastolic blood pressure at admission,laboratory test indicators at admission(blood glucose,glycosylated hemoglobin,homocysteine,total cholesterol,triglyceride,low-density lipoprotein cholesterol,high-density lipoprotein cholesterol,neutrophils,lymphocytes and neutrophil-to-lymphocyte ratio),responsible occlusion artery(internal carotid artery,middle cerebral artery,anterior cerebral artery),affected cerebral hemisphere,collateral circulation score,and medications used within 72 h after admission(intravenous thrombolysis+dual antiplatelet therapy,tirofiban+dual antiplatelet therapy,argatroban+dual antiplatelet therapy,argatroban alone,dual antiplatelet therapy alone).Variables with statistically significant differences in univariate analysis were included in multivariate Logistic regression analysis to explore the independent influencing factors for END in patients with acute anterior circulation large-vessel occlusive mild stroke treated with medications alone.Results A total of 208 patients with acute anterior circulation large-vessel occlusive mild stroke were included,with 143 males and 65 females,aged 38-85 years,with an average age of(64±9)years.Among them,86 patients were in the neurological deterioration group and 122 in the stable condition group.(1)There were statistically significant differences between the neurological deterioration group and the stable condition group in terms of history of diabetes mellitus(39.5％[34/86]vs.17.2％[21/122]),smoking history(43.0％[37/86]vs.29.5％[36/122]),left cerebral hemisphere lesion(57.0％[49/86]vs.41.0％[50/122]),collateral circulation score(4[3,5]vs.5[4,5]),time from onset to admission(7.0[3.0,17.0]hvs.4.3[2.0,11.0]h),blood glucose at admission(7.4[5.8,10.0]mmol/L vs.6.7[5.8,7.7]mmol/L),neutrophil-to-lymphocyte ratio(3.8[2.4,5.1]vs.3.0[2.1,4.3]),dual antiplatelet therapy alone(19.8％[17/86]vs.6.6％[8/122]),and argatroban+dual antiplatelet therapy(8.1％[7/86]vs.29.5％[36/122];all P＜0.05).There were no statistically significant differences in the results of the remaining univariate analyses(all P＞0.05).(2)Multivariate Logistic regression analysis showed that diabetes mellitus(OR,2.674,95％CI 1.121-6.377,P=0.027)and left cerebral hemisphere vessel occlusion(OR,2.030,95％CI I.083-3.806,P=0.027)were independent risk factors for END in acute anterior circulation large-vessel occlusive mild stroke.Argatroban+dual antiplatelet therapy(OR,0.267,95％CI 0.116-0.613,P=0.002)and high collateral circulation score(OR,0.551,95％CI 0.368-0.824,P=0.004)were independent protective factors for END in acute anterior circulation large-vessel occlusive mild stroke.Conclusions Acute anterior circulation large-vessel occlusive mild stroke patients with diabetes mellitus or left cerebral hemisphere lesions are prone to END.The combination of argatroban and dual antiplatelet therapy and good collateral circulation can reduce the risk of END.

Objective To investigate the standardized use of anti-tumor drugs in Beijing hospitals in 2024,and put forward relevant policy suggestions on standardizing the use of anti-tumor drugs.Methods In 2024,Beijing Cancer Quality Control Center organized internal medicine and pharmacy experts to conduct a research on the standardized use of anti-tumor drugs in 556 medical records from 60 hospitals.The evaluation content included pathological diagnosis and tumor staging,treatment process and quality control.Results The evaluation of the use of anti-tumor drugs in 60 hospitals in Beijing found that the main problems of non-standardized use of anti-tumor drugs were incomplete or unsigned informed consent forms;incomplete,un-staged or incorrect staging of tumors;lack of adverse reaction evaluation;incomplete records of drug application on the day of treatment;no reasons given for adjustments in drug dosage and administration methods,and incomplete efficacy evaluation.Conclusion It is necessary to strengthen the training and guidance of medical staff,carry out regular quality control inspections,enhance information construction,establish a multi-disciplinary diagnosis and treatment system,and establish a regular follow-up mechanism,so as to further standardize and improve the standard rate of antitumor drug use in hospitals.

Objective To investigate the standardized use of anti-tumor drugs in Beijing hospitals in 2024,and put forward relevant policy suggestions on standardizing the use of anti-tumor drugs.Methods In 2024,Beijing Cancer Quality Control Center organized internal medicine and pharmacy experts to conduct a research on the standardized use of anti-tumor drugs in 556 medical records from 60 hospitals.The evaluation content included pathological diagnosis and tumor staging,treatment process and quality control.Results The evaluation of the use of anti-tumor drugs in 60 hospitals in Beijing found that the main problems of non-standardized use of anti-tumor drugs were incomplete or unsigned informed consent forms;incomplete,un-staged or incorrect staging of tumors;lack of adverse reaction evaluation;incomplete records of drug application on the day of treatment;no reasons given for adjustments in drug dosage and administration methods,and incomplete efficacy evaluation.Conclusion It is necessary to strengthen the training and guidance of medical staff,carry out regular quality control inspections,enhance information construction,establish a multi-disciplinary diagnosis and treatment system,and establish a regular follow-up mechanism,so as to further standardize and improve the standard rate of antitumor drug use in hospitals.

Objective To obtain the range of anatomical parameters of healthy knee joints in Chinese males and validate a statistical shape model(SSM)based on the geometric shape of a healthy knee to provide references for the design of knee SSM-based prostheses.Methods Computed tomography(CT)images of knee joints from 112 healthy males were acquired to build three-dimensional(3D)knee joint models.Each model was the target model separately,and the remaining models were used as the training set for principal component analysis(PCA).The obtained knee SSM was fitted to the target model to predict the SSM.The exact anatomical measurement points were marked on the sample and SSM prediction models,and 17 linear and 3 angular parameters were derived.The values of the anatomical parameters were statistically tested using an independent samples t-test and Mann-Whitney U-test,and the validity of the SSM in terms of geometric shape was demonstrated if the resulting P-values were all greater than 0.05.Results Qualitative and quantitative comparative analyses of anatomical parameters showed that the mean deviation of linear parameters was less than 6 mm,and that of angular parameters was less than 2.5°.The results of statistical tests showed P＞0.05 for all anatomical parameters,proving that the knee SSM prediction model was not statistically different from the true healthy model in terms of geometric shape.Conclusions This study derived a reference range of anatomical parameters for a healthy knee and demonstrated that the knee SSM model was consistent with the real healthy model in terms of shape.The results provide a reference for the design of knee SSM-based prostheses.

A 72-year-old female patient was admitted to the emergency department of Qintang District People′s Hospital of Guigang City in August 2023 due to chills and fever, abdominal distension and pain, diarrhea, cough and shortness of breath for 1 day. She had a history of chronic obstructive and pulmonary heart disease, stage Ⅲ hypertension, and ceftazidime allergy. Clinical diagnosis of acute bacterial infection of chronic obstructive pneumonia was made and levofloxacin combined with piperacillin/tazobactam were given as symptomatic treatment. The blood culture reported Campylobacter fetus after four days, and the patient was cured and discharged after seven days with negative blood culture. The morphology and mass spectrometry identification of the strain were consistent with the definition of Campylobacter fetus. Whole genome sequencing predicted the multi-site sequence type as Campylobacter fetus subsp. fetus( Cff) ST20, carrying the tetracycline resistance gene tet (O/M/O), 18 flagella genes (including rpoN gene from Campylobacter jejuni. these genes were not found in the other two Campylobacter fetus subspecies), and six virulence genes (including like-typhoidal toxin and typhoid toxin genes). The pathogen has the ecological characteristics of parasitic farmed animal colonization and the biological characteristics of high mobility and virulence. These attributes facilitated its entry into the bloodstream via the fecal-oral route, leading to invasive infections.