This study was aimed at investigating differentially expressed genes(DEGs)in Clonorchis sinensis(C.sinensis)meta-cercariae and newly excysted juveniles(NEJs)cultured in vitro for 1 hour or 3 hours,through transcriptomic analysis.Our objective was to explore the mechanisms underlying host invasion.Metacercariae were digested and isolated from Pseudorasbora parva infected with C.sinensis.The metacercariae excysted and developed into NEJs in vitro.Subsequently,the mRNA of metacercariae and NEJs cultured in vitro for 1 hour or 3 hours was extracted for transcriptomic sequencing analysis to screen for DEGs,and to conduct GO and KEGG analyses.A protein-protein interaction network(PPI)was constructed to identify hub genes.A total of 1 218 DEGs were de-tected.The main enriched GO terms of DEGs included transcription regulator activity and gated channel activity(primarily K+).The KEGG pathways significantly enriched in DEGs included cholesterol metabolism,lysosome,synthesis,secretion,and action of para-thyroid hormone.ZFAND4-2,BIRC6,and other genes were screened and identified as hub genes through PPI network analysis.Addi-tionally,abundant differential expression of cathepsin-related genes,including Cathepsin L and Cathepsin F,were observed before and after excystment in C.sinensis.Therefore,significant transcriptional level changes occurred in the metacercariae of C.sinensis be-fore and after excystation,and enrichment was observed primarily in signaling pathways,such as activation of growth and material me-tabolism,that regulate parasite growth and development.Meanwhile,biological events conducive to parasite invasion,migration,and adhesion were triggered.

Objective:To investigate the application of time series models in forecasting pre-hospital emergency ambulance trips in Handan City and develop the LPro ensemble model for improved prediction accuracy to support emergency resource allocation.Methods:Pre-hospital emergency data from Handan Emergency Medical Command Center (2019-2023) were retrospectively analyzed. From 324 799 original records, 289 949 valid records were included after cleaning. The training set (2019-2022: 215 918 records) included 35 527 records in 2019, 52 015 in 2020, 61 836 in 2021, and 66 540 in 2022. The validation set (2023) contained 74 031 records. ARIMA, linear trend seasonal, exponential smoothing, and Prophet models were fitted to the training set. The LPro ensemble model was constructed using MAPE-based weighting (linear trend seasonal model: 0.38, Prophet: 0.62). Performance metrics included MAPE, RMSE, MAE, and R 2. Results:Data showed annual growth (compound annual growth rate 23.27%) and seasonal patterns (October peaks, February troughs). Ambulance dispatches increased annually with monthly cyclical patterns. For 2023 validation predictions: ARIMA (MAPE 8.76%, RMSE 619, MAE 491, R 2 0.4563), linear trend seasonal (MAPE 9.83%, RMSE 671, MAE 545, R 2 0.3608), Prophet (MAPE 8.43%, RMSE 562, MAE 503, R 2 0.5513), exponential smoothing (MAPE 8.08%, RMSE 643, MAE 410, R 2 0.4124). LPro model showed superior performance (MAPE 7.05%, RMSE 491, MAE 393, R 2 0.6570), with 16.37% lower MAPE, 12.63% lower RMSE, 21.87% lower MAE, and 19.17% higher R 2 versus Prophet. Conclusion:The LPro ensemble model substantially enhances prediction accuracy and reliability, offering scientific support for emergency resource optimization and dispatch scheduling in Handan City.

Objective To study the 50％effective dose(ED50)and ED95 of remimazolam in the induc-tion of general anesthesia in preschool children,and to observe its safety and efficacy.Methods Children un-dergoing elective surgery who received endotracheal intubation and general anesthesia from March to October 2024 were selected as the research objects.The ED50 and ED95 of remimazolam were calculated by the modified sequential method.On this basis the additional,the aditional 68 subjects were randomly divided into the remimazolam group(group R,n=34)and the propofol group(group P,n=34).The respiratory rate(RR)and bispectral index(BIS)before induction(T0),1 min(T1)and 3 min(T2)after administration of remima-zolam or propofol in each group were recorded,and the oxygen saturation(SpO2),heart rate(HR)and mean arterial pressure(MAP)at T0,T1,T2,and 1 min after tracheal intubation(T3)were recorded.The success rate of sedation induction,hemodynamic stability and the incidence of adverse reactions such as injection pain,bradycardia,hypotension and panic during induction were compared between the two groups.Results Probit regression analysis showed that the ED50 of remimazolam for inducing loss of consciousness in preschool chil-dren was 0.414 mg/kg,95％CI:0.381-0.448,and the ED95 was 0.474 mg/kg,95％CI:0.443-0.826.The success rate of sedation induction in both groups was 100％.The disappearance time of eyelash reflex in group R was(59.44±5.24)s,which was longer than(29.79±4.04)s in group P(P＜0.001).The RR and BIS of group R at T1 and T2 were higher than those of group P,and the HR and MAP of group R at T1,T2 and T3 were also higher than those of group P(P＜0.05).The incidence of injection pain,bradycardia and hypoten-sion in group R was lower than that in group P(P＜0.05).There was 1 case of panic in group R.Conclusion The ED50 and ED95 of remimazolam for loss of consciousness during general induction in preschool children are 0.414 mg/kg and 0.474 mg/kg respectively.It has a definite sedative effect,stable hemodynamics and a low incidence of adverse reactions for the induction of general anesthesia in this population,demonstrating its suit-ability for clinical use.

Objective To discuss the curative effect of hepatic arterial infusion chemotherapy(HAIC)combined with programmed death receptor-1(PD-1)inhibitors and anti-angiogenesis therapy for BCLC stage C hepatocellular carcinoma(HCC).Methods The patients with HCC of BCLC stage C,who received HAIC combined with PD-1 inhibitors and anti-angiogenesis therapy at the First Affiliated Hospital of Soochow University of China from January 2021 to December 2023,were collected for this study.The time from the start of treatment to complete remission(CR)of disease,the dynamic changes in alpha-fetoprotein(AFP)levels and the recurrence during follow-up period were analyzed.The relevant literature of the above triple regimen for HCC was searched from PubMed database and its curative effect was analyzed.Results Of the 214 HCC patients treated with triple therapy regimen,9(4.2％)achieved CR.The time from the start of treatment to CR was 2-10 months.The patients were followed up for 5-20 months.The time of AFP level returning to normal value was from 79 to 259 days.One patient developed recurrence 10 months after CR,and the other 8 patients maintained the CR status.A total of 12 articles were retrieved,the reported CR rates ranged from 0 to 16.5％.Conclusion A CR can be achieved in a few patients with advanced HCC treated with HAIC combined with PD-1 inhibitors and anti-angiogenesis therapy.

Objective:To explore the reliability and validity of the Chinese version of Hammersmith Neonatal Neurological Examination (C-HNNE).Methods:A prospective cohort study.One hundred and fourteen neonates born in Longgang District Maternity & Child Healthcare Hospital of Shenzhen City between October 2022 and August 2023, who were hospitalized in the Neonatology or Obstetrics Department after birth and met the inclusion criteria, were enrolled as study subjects.They were divided into an early preterm group(34 cases), a mid-late preterm group(50 cases), and a full-term group(30 cases) based on gestational age.The first C-HNNE assessment was completed within 24 hours after birth, and 20 cases from each group were selected for inter-rater reliability assessment; 48 hours after the first C-HNNE assessment, 20 cases from each group were selected to undergo the C-HNNE assessment again for test-retest reliability assessment.At corrected 4 months of age, short-term neurodevelopmental outcomes were determined by pediatric rehabilitation physicians using clinical examination combined with general movements (GMs) assessment.Inter-rater reliability and test-retest reliability were assessed by calculating intra-class correlation coefficient (ICC). The optimal cutoff scores of the C-HNNE for each group were determined by plotting receiver operating characteristic(ROC) curves.Predictive validity was determined by calculating sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV).Results:The highest sensitivity for predicting neonatal corrected neuromotor developmental outcome at 4 months of age was achieved when the optimal C-HNNE scores were taken as 22.25, 25.25, and 29.25 in the early preterm, mid-late preterm, and term groups, respectively.The ICCs for the inter-rater reliability of the total C-HNNE score and subscale scores in all 3 groups were above 0.7, indicating good reliability.The ICCs for test-retest reliability of some individual items were<0.6, indicating moderate reliability.In terms of validity, the correlation coefficients between the total C-HNNE scores and the GMs scores of the three groups were 0.550, 0.483, 0.473 (all P<0.01), and the sensitivity of the C-HNNE for predicting neurodevelopmental developmental outcomes in neonates corrected to 4 months of age was 82.9%, specificity was 70.4%, PPV was 58.0%, and NPV was 89.3%. Conclusions:C-HNNE demonstrates good reliability and validity and can be used as a routine bedside examination program for early neonatal life.

This study was aimed at investigating differentially expressed genes(DEGs)in Clonorchis sinensis(C.sinensis)meta-cercariae and newly excysted juveniles(NEJs)cultured in vitro for 1 hour or 3 hours,through transcriptomic analysis.Our objective was to explore the mechanisms underlying host invasion.Metacercariae were digested and isolated from Pseudorasbora parva infected with C.sinensis.The metacercariae excysted and developed into NEJs in vitro.Subsequently,the mRNA of metacercariae and NEJs cultured in vitro for 1 hour or 3 hours was extracted for transcriptomic sequencing analysis to screen for DEGs,and to conduct GO and KEGG analyses.A protein-protein interaction network(PPI)was constructed to identify hub genes.A total of 1 218 DEGs were de-tected.The main enriched GO terms of DEGs included transcription regulator activity and gated channel activity(primarily K+).The KEGG pathways significantly enriched in DEGs included cholesterol metabolism,lysosome,synthesis,secretion,and action of para-thyroid hormone.ZFAND4-2,BIRC6,and other genes were screened and identified as hub genes through PPI network analysis.Addi-tionally,abundant differential expression of cathepsin-related genes,including Cathepsin L and Cathepsin F,were observed before and after excystment in C.sinensis.Therefore,significant transcriptional level changes occurred in the metacercariae of C.sinensis be-fore and after excystation,and enrichment was observed primarily in signaling pathways,such as activation of growth and material me-tabolism,that regulate parasite growth and development.Meanwhile,biological events conducive to parasite invasion,migration,and adhesion were triggered.

Objective:To explore the reliability and validity of the Chinese version of Hammersmith Neonatal Neurological Examination (C-HNNE).Methods:A prospective cohort study.One hundred and fourteen neonates born in Longgang District Maternity & Child Healthcare Hospital of Shenzhen City between October 2022 and August 2023, who were hospitalized in the Neonatology or Obstetrics Department after birth and met the inclusion criteria, were enrolled as study subjects.They were divided into an early preterm group(34 cases), a mid-late preterm group(50 cases), and a full-term group(30 cases) based on gestational age.The first C-HNNE assessment was completed within 24 hours after birth, and 20 cases from each group were selected for inter-rater reliability assessment; 48 hours after the first C-HNNE assessment, 20 cases from each group were selected to undergo the C-HNNE assessment again for test-retest reliability assessment.At corrected 4 months of age, short-term neurodevelopmental outcomes were determined by pediatric rehabilitation physicians using clinical examination combined with general movements (GMs) assessment.Inter-rater reliability and test-retest reliability were assessed by calculating intra-class correlation coefficient (ICC). The optimal cutoff scores of the C-HNNE for each group were determined by plotting receiver operating characteristic(ROC) curves.Predictive validity was determined by calculating sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV).Results:The highest sensitivity for predicting neonatal corrected neuromotor developmental outcome at 4 months of age was achieved when the optimal C-HNNE scores were taken as 22.25, 25.25, and 29.25 in the early preterm, mid-late preterm, and term groups, respectively.The ICCs for the inter-rater reliability of the total C-HNNE score and subscale scores in all 3 groups were above 0.7, indicating good reliability.The ICCs for test-retest reliability of some individual items were<0.6, indicating moderate reliability.In terms of validity, the correlation coefficients between the total C-HNNE scores and the GMs scores of the three groups were 0.550, 0.483, 0.473 (all P<0.01), and the sensitivity of the C-HNNE for predicting neurodevelopmental developmental outcomes in neonates corrected to 4 months of age was 82.9%, specificity was 70.4%, PPV was 58.0%, and NPV was 89.3%. Conclusions:C-HNNE demonstrates good reliability and validity and can be used as a routine bedside examination program for early neonatal life.

BACKGROUND:Traditional Chinese medicine compound prescription has a long history in the treatment of primary osteoporosis,and the curative effect is definite,but the medication rule and mechanism are not clear. OBJECTIVE:Using the methodology of data mining and network pharmacology,to explore and verify the law of drug use and molecular mechanism of modern traditional Chinese medicine in the treatment of primary osteoporosis. METHODS:The relevant documents included in CNKI,WanFang,VIP and PubMed were used as data sources,and the relevant data were statistically counted and extracted by Microsoft EXCEL2019,IBMSPSS25.0 and other software.The high-frequency drugs obtained from the data statistics were analyzed by association rules analysis and cluster analysis,and the core drug combination of traditional Chinese medicine compound prescription in the treatment of primary osteoporosis was obtained by combining the two results.The therapeutic mechanism of this combination was explained by network pharmacology and verified by molecular docking. RESULTS AND CONCLUSION:Finally,151 articles were included and 207 prescriptions were selected,involving 285 flavors of Chinese herbs.(1)Ten groups of important drug combinations were obtained through the above two analyses,among which the core drug combination with the highest confidence and improvement was"Drynaria-Eucommia-Angelica."The key components of the combination in the treatment of primary osteoporosis were quercetin,kaempferol,naringenin and so on.The core targets were SRC proto-oncogene,phosphoinositide-3-Kinase regulatory subunit 1 and RELA proto-oncogene.The main pathways were cancer signaling pathway,JAK-STAT signaling pathway,VEGF signaling pathway,and NF-κB signaling pathway.(2)The key active components were docked with the core targets,and the two showed a good combination.To conclude,Chinese herbal compound therapy in the treatment of primary osteoporosis can use a variety of active components to exert its efficacy through multiple signal pathways and acting on multiple targets,which can provide a theoretical basis for the research and development of new drugs for the follow-up treatment of primary osteoporosis.

Objective:To assess if metagenomic next-generation sequencing (mNGS) of periprosthetic joint tissue can provide an alternative rapid and sensitive tool for the diagnosis of prosthetic joint infection (PJI), especially compared to microbiological culture.Methods:A total of 33 eligible patients who underwent revision arthroplasty from June 2019 to June 2020 in orthopedic surgery department of the first affiliated hospital of Zhengzhou University were retrospectively analyzed. Twenty-one patients were included in PJI group according to the American Academy of Musculoskeletal Infection diagnostic criteria, with 17 cases of knee and 4 cases of hip, including 9 cases of male and 12 cases of female, with an average age of 59.14±14.55 years old (range from 28 to 84), and an average BMI of 23.7±2.8 kg/m 2 (range from 17.7 to 29.4 kg/m 2). Twelve patients were included in aseptic loosening group (control group), with 4 cases of knee and 8 cases of hip, including 4 cases of male and 8 cases of female, with an average age of 53.08±10.05 years old (range from 39 to 70), and an average BMI of 25.2±2.9 kg/m 2 (range from 18.3 to 31.2 kg/m 2 ). Microbiological culture results of synovial fluid and periprosthetic joint tissue and mNGS results of periprosthetic joint tissue were collected. The sensitivity and specificity of mNGS and microbiological culture were calculated and compared. The species of pathogenic microorganismsdetected by the two techniques were summarized. In addition, the impact of antibiotic use on the efficacy of both techniques were compared. Results:mNGS detected 13 positive cases and microbiological culture detected 6 positive cases in the PJI group. In the aseptic loosening group, 1 case was determined positive by mNGS, and all the microbiological culture results were negative. In the diagnosis of PJI, mNGS showed significantly higher sensitivity than that of culture (61.9% vs 28.6%, χ2=4.71, P=0.03), while no statistical difference was observed in terms of specificity (91.7% vs 100%, χ2=1.04, P=0.31). In the PJI cases with prior exposure to antibiotics within two weeks, the sensitivity of mNGS was significantly higher than that of culture (53.8% vs 15.4%, χ2=4.25, P=0.04). However, there was no significant difference in the sensitivity between mNGS and culture in patients without antibioticsexposure (66.7% vs 44.4%, χ2=0.90, P=0.34). In the detection of pathogenic microorganism, mNGS detected 9 kinds of bacteria (Staphylococcus aureus, Staphylococcus family, Moraxella Oslo, Propionibacterium acnes, Streptococcus acnes, Staphylococcus epidermidis, Mycobacterium tuberculosis, Staphylococcus Lyons, Bacteroides fragilis) and 2 kinds of fungi (Aspergillus fumigatus, Candida parapsilosis), while microbiological culture detected 3 kinds of bacteria (Staphylococcus aureus, Moraxella catarrhalis, Mycobacterium tuberculosis) and one kind of fungi (Candida parapsilosis). mNGS and microbiological culture were both positive in 5 cases, among which 3 cases had completely matched results (Staphylococcus aureus, Mycobacterium tuberculosis, Candida parapsilosis), one case had partly matched results (mNGS detected more bacteria than culture) and one case had totally mismatched results. Additionally, in the diagnosis of the 3 included tuberculous PJI, mNGS showed 100% specificity and sensitivity. Conclusion:mNGS of periprosthetic joint tissue is a more powerful tool for diagnosis and pathogen detection of PJI compared to microbiological culture, especially in the diagnosis of tuberculosis PJI. Besides, mNGS is more resistant to antibiotic exposure than culture.

Objective To explore the clinical application value of serum tumor abnormal protein (TAP)in early diagnosis of breast cancer.Methods The serum TAP level was determined in 53 hospitalized patients with breast cancer and 65 cases of normal physical examination population as the control group.We further compared the positive rate of TAP in the two groups and the expression level of TAP between different clinical pathological parameters in breast cancer group.Results There was no case of TAP positive in the control group,while the positive rate of TAP was as high as 83.02％ in breast cancer group.TAP positive rate of the patients with negative PR (100.00％) was significantly higher than that of PR positive patients (73.53％)(P=0.019).However there was no significant difference of TAP positive rate between patients with different ages,clinical stages,lymph node metastasis and the different expression of ER,C-erbB2 and Ki67.Conclusion It might be clinically valuable to use TAP expression level as a screening marker for breast cancer in combination with the breast cancer hormone PR.