In this study,the carboxylated magnetic beads were coupled with bivalent nanobodies a-gainst porcine epidemic diarrhea virus(PEDV)M protein to construct immunomagnetic beads(IM-NBs-Ⅱ),The capture and enrichment function of IMNBs-Ⅱ was verified by using PEDV propaga-ted in Vero cells.A one-step RT-qPCR detection method for PEDV was established by combining the characteristics of IMNBs-Ⅱ with the detection advantages of reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR).Specific analysis found that this method has no cross reactivity with swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,porcine circovirus,indicating that it has good specificity.Sensitivity analysis re-vealed that the detection sensitivity of the RT-qPCR based on IMNBs-Ⅱ was increased 10 times compared to traditional RT-qPCR methods.Detection of the clinical samples confirm that the RT qPCR method based on IMNBs-Ⅱ is suitable for rapid and accurate detection of clinical feces and tissue samples.The method established in this study effectively avoids contamination issues during nucleic acid extraction,simplifies experimental procedures,and saves detection time,which pro-vides a method for efficient detection of PEDV.

Objective To explore the effect and mechanism of acupuncture at Zusanli(ST36)on inflammatory response in mice with mesenteric lymphadenitis based on the mitogen-activated protein ki-nase(MAPK)/nuclear factor-KB(NF-κB)signaling pathway.Methods Twelve male Kunming strain mice were randomly selected as blank control group,and mice with successfully established mes-enteric lymphadenitis models were randomly divided into model group and acupuncture at Zusanli group,with 12 mice in each group.Mice in the acupuncture at Zusanli group received electroacupunc-ture treatment at Zusanli for 4 days,while those in the blank control group and the model group re-ceived no treatment.Behavioral characteristics of mice in each group were observed.Hematoxylin and eosin(HE)staining was used to evaluate pathological changes in mesenteric lymph node tissues of mice.Enzyme-linked immunosorbent assay(ELISA)was employed to detect the expression levels of pro-inflammatory cytokines[tumor necrosis factor-α(TNF-α),interleukin-1 β(IL-1β),interleu-kin-6(IL-6)]in mouse serum.Western blot was used to detect the expression of autophagy-related proteins(Beclin1,LC3-Ⅱ,LC3-Ⅰ,p62)and MAPK/NF-κB signaling pathway-related proteins(p38 MAPK,p-p38 MAPK,NF-κB p65,p-NF-κB p65)in mesenteric lymph node tissues of mice.Results Mice in the model group exhibited decreased food intake,lethargy,soft and loose stools,and occasional kyphosis.The behavioral characteristics of mice in the acupuncture at Zusanli group were improved compared with those in the model group.HE staining results showed pathological changes in mesenteric lymph node tissues of mice in the model group,while the pathological changes in the acupuncture at Zusanli group were improved compared with those in the model group.ELISA results showed that the levels of TNF-α,IL-1β,and IL-6 in serum of mice in the model group were higher than those in the blank control group,while those in the acupuncture at Zusanli group were lower than those in the model group(P＜0.05).Western blot results showed that the relative ex-pression of Beclin1 protein and the LC3-Ⅱ/LC3-Ⅰ in mesenteric lymph node tissues of mice in the model group were lower than those in the blank control group,while the relative expression of p62 protein and the ratios of p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 were higher than those in the blank control group(P＜0.05).In the acupuncture at Zusanli group,the relative ex-pression of Beclin1 protein and the LC3-Ⅱ/LC3-Ⅰ were higher than those in the model group,while the relative expression of p62 protein and p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 were lower than those in the model group(P＜0.05).Conclusion Acupuncture at Zusanli alleviates the inflammatory response in mice with mesenteric lymphadenitis by inhibiting the p38 MAPK/NF-κB signaling pathway.

Objective To explore the prognosis-related genes of lung adenocarcinoma(LUAD)and establish a prognostic prediction model for LUAD.Methods The differentially expressed genes of LUAD tissues and adjacent normal tissues in the GSE43458,GSE7670,and GSE140797 datasets were screened.The protein-protein interaction(PPI)network analysis,gene ontology(GO)function,Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analyses,least absolute shrinkage and selection operator for prognosis coefficient screening,disease-specific survival analysis,Cox regression analysis,and gene correlation analysis were performed.Independent prognostic genes of LUAD were screened from the differential genes,and a prognostic prediction model of LUAD was established.The expression of independent prognostic genes was analyzed,and the predictive model was evaluated by receiver operating characteristic(ROC)curve.The GSE68465 data set was used as an external validation set to verify the predictive model.Results There were 197 up-regulated differential genes and 77 down-regulated differential genes in LUAD and adjacent normal tissues common to the three datasets.Through stepwise screening,two genes,IL7R and SLC2A1,were identified as independent prognostic factors for LUAD.IL7R was an independent protective factor,while the SLC2A1 was an independent risk factor.A prediction model for curve was built with IL7R and SLC2A1.The prediction model for LUAD constructed with IL7R and SLC2A1 is as follows:Risk score=(-0.694)×expression level of IL7R+0.763×expression level of SLC2A1.Conclusion This study screened out IL7R as an independent prognostic protective factor for LUDA,and SLC2A1 as an independent prognostic risk factor for LUAD.The LUDA prediction model constructed based on these two genes has good predictive ability and generalization ability,which can provide references for the research and clinical individualized treatment of LUAD.

Objective:To evaluate the differences in total effectiveness, Visual Analogue Scale (VAS) scores, and Cox Menstrual Symptom Scale (CMSS) scores among four non-invasive Traditional Chinese Medicine (TCM) techniques: moxibustion, massage therapy, acupoint application, and auricular acupressure, for the care of primary dysmenorrhea (PD) using network Meta-analysis, providing evidence-based guidance for clinical decision-making.Methods:Randomized controlled trials (RCTs) involving the four non-invasive TCM techniques for PD care were retrieved from databases including China National Knowledge Infrastructure, VIP, Wanfang, SinoMed, PubMed, Embase, Web of Science, and Cochrane Library up to November 30, 2023. Network Meta-analysis was conducted using RevMan 5.4 and Stata 17.0.Results:A total of 83 studies involving 8 370 PD patients were included. The network Meta-analysis showed that all four TCM techniques were superior to control measures in terms of total effectiveness and reducing VAS scores. Moxibustion, acupoint application, and auricular acupressure were also superior in reducing CMSS scores. Ranking of the effects of different interventions: total effectiveness: auricular acupressure > moxibustion > acupoint application > massage therapy > acupuncture > Chinese patent medicine > Western medicine > blank control > placebo treatment; VAS scores: moxibustion > auricular acupressure > acupoint application > acupuncture > placebo treatment > massage therapy > Western medicine > Chinese patent medicine > blank control; CMSS scores: auricular acupressure > moxibustion > acupuncture > placebo treatment > acupoint application > Western medicine > blank control.Conclusions:The four non-invasive TCM techniques have significant advantages in PD care, with auricular acupressure and moxibustion being the preferred choices for clinical care.

In this study,the carboxylated magnetic beads were coupled with bivalent nanobodies a-gainst porcine epidemic diarrhea virus(PEDV)M protein to construct immunomagnetic beads(IM-NBs-Ⅱ),The capture and enrichment function of IMNBs-Ⅱ was verified by using PEDV propaga-ted in Vero cells.A one-step RT-qPCR detection method for PEDV was established by combining the characteristics of IMNBs-Ⅱ with the detection advantages of reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR).Specific analysis found that this method has no cross reactivity with swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,porcine circovirus,indicating that it has good specificity.Sensitivity analysis re-vealed that the detection sensitivity of the RT-qPCR based on IMNBs-Ⅱ was increased 10 times compared to traditional RT-qPCR methods.Detection of the clinical samples confirm that the RT qPCR method based on IMNBs-Ⅱ is suitable for rapid and accurate detection of clinical feces and tissue samples.The method established in this study effectively avoids contamination issues during nucleic acid extraction,simplifies experimental procedures,and saves detection time,which pro-vides a method for efficient detection of PEDV.

Objective:To evaluate the differences in total effectiveness, Visual Analogue Scale (VAS) scores, and Cox Menstrual Symptom Scale (CMSS) scores among four non-invasive Traditional Chinese Medicine (TCM) techniques: moxibustion, massage therapy, acupoint application, and auricular acupressure, for the care of primary dysmenorrhea (PD) using network Meta-analysis, providing evidence-based guidance for clinical decision-making.Methods:Randomized controlled trials (RCTs) involving the four non-invasive TCM techniques for PD care were retrieved from databases including China National Knowledge Infrastructure, VIP, Wanfang, SinoMed, PubMed, Embase, Web of Science, and Cochrane Library up to November 30, 2023. Network Meta-analysis was conducted using RevMan 5.4 and Stata 17.0.Results:A total of 83 studies involving 8 370 PD patients were included. The network Meta-analysis showed that all four TCM techniques were superior to control measures in terms of total effectiveness and reducing VAS scores. Moxibustion, acupoint application, and auricular acupressure were also superior in reducing CMSS scores. Ranking of the effects of different interventions: total effectiveness: auricular acupressure > moxibustion > acupoint application > massage therapy > acupuncture > Chinese patent medicine > Western medicine > blank control > placebo treatment; VAS scores: moxibustion > auricular acupressure > acupoint application > acupuncture > placebo treatment > massage therapy > Western medicine > Chinese patent medicine > blank control; CMSS scores: auricular acupressure > moxibustion > acupuncture > placebo treatment > acupoint application > Western medicine > blank control.Conclusions:The four non-invasive TCM techniques have significant advantages in PD care, with auricular acupressure and moxibustion being the preferred choices for clinical care.

Objective To explore the prognosis-related genes of lung adenocarcinoma(LUAD)and establish a prognostic prediction model for LUAD.Methods The differentially expressed genes of LUAD tissues and adjacent normal tissues in the GSE43458,GSE7670,and GSE140797 datasets were screened.The protein-protein interaction(PPI)network analysis,gene ontology(GO)function,Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analyses,least absolute shrinkage and selection operator for prognosis coefficient screening,disease-specific survival analysis,Cox regression analysis,and gene correlation analysis were performed.Independent prognostic genes of LUAD were screened from the differential genes,and a prognostic prediction model of LUAD was established.The expression of independent prognostic genes was analyzed,and the predictive model was evaluated by receiver operating characteristic(ROC)curve.The GSE68465 data set was used as an external validation set to verify the predictive model.Results There were 197 up-regulated differential genes and 77 down-regulated differential genes in LUAD and adjacent normal tissues common to the three datasets.Through stepwise screening,two genes,IL7R and SLC2A1,were identified as independent prognostic factors for LUAD.IL7R was an independent protective factor,while the SLC2A1 was an independent risk factor.A prediction model for curve was built with IL7R and SLC2A1.The prediction model for LUAD constructed with IL7R and SLC2A1 is as follows:Risk score=(-0.694)×expression level of IL7R+0.763×expression level of SLC2A1.Conclusion This study screened out IL7R as an independent prognostic protective factor for LUDA,and SLC2A1 as an independent prognostic risk factor for LUAD.The LUDA prediction model constructed based on these two genes has good predictive ability and generalization ability,which can provide references for the research and clinical individualized treatment of LUAD.

Objective To analyze the curative effect of acupuncture at Gongsun, Sifeng combined with visceral acupoint tapping in children with mesenteric lymphadenitis (type of gastrointestinal heat accumulation), and its influence on serum intestinal fatty acid-binding protein (I-FABP) and vasoactive intestinal peptide (VIP). Methods A total of 60 children with mesenteric lymphadenitis (type of gastrointestinal heat accumulation) were selected, and were divided into control group (n=30, conventional treatment combined with acupuncture at Gongsun and Sifeng acupoints) and study group (n=30, visceral acupoint tapping on the basis of the control group) according to the random number table method. The clinical efficacy, changes of main symptoms and signs were compared, and the expression levels of serum I-FABP and VIP were detected. Results The total effective rate of the study group was higher than that of the control group(P < 0.05). After 2 weeks of treatment, the serum I-FABP levels of the two groups were lower than that before treatment, and the serum I-FABP level of the study group was lower than that of the control group (P < 0.05). After 2 weeks of treatment, the serum VIP levels of the two groups were higher than that before treatment, and the serum VIP level of the study group was higher than that of the control group (P < 0.05). After 2 weeks of treatment, the improvement of main symptoms and signs in the study group was better than that in the control group (P < 0.05). Conclusion Acupuncture at Gongsun and Sifeng combined with visceral acupoint tapping can be used to treat children with mesenteric lymphadytis (gastrointestinal heat accumulation type), regulate and improve gastrointestinal function, adjust the level of serum I-FABP and VI, and relieve the main symptoms and signs of children, and can be used as an effective therapeutic method.

Objective:To analyze the configuration characteristics, service utilization trend and outcome of preimplantation genetic diagnosis (PGD) in China.Methods:A longitudinal study and a cross-sectional survey in 2020 were used to analyze the number of PGD facilities and the trend of service utilization in China from 2004 to 2020. The characteristics of 2020 technology configuration were described from different geographical distribution, city positioning, and organization categories. The number of couples treated, the number of egg retrieval cycles and the number of diagnostic cycles were used to evaluate the scale of the service.Results:From 2004 to 2020, the number of PGD institutions increased from only 4 (10.8%) in 2004 to 78 (19.0%) in 2020. Although the number of PGD cycles increased with it, the overall proportion was not high, ranging from 0.3% to 4.1%. A total of 21 241 PGD cycles were reported in 2020, accounting for 4.1% of the total number of egg retrievals. The PGD technology configuration in 2020 was mainly characterized by provincial capitals [82.1% (64/78)], public hospitals [94.9% (74/78)], and comprehensive hospitals [59.0% (46/78)]. There were significant differences in the median of the number of PGD egg retrieval cycles in eastern, central and western institutions, which were 203.0, 177.5 and 79.0, respectively. In 2020, the PGD clinical pregnancy rate was 60.2% (10 326/17 158), the delivery rate was 51.1% (8 767/17 158), the multiple delivery rate was 1.9% (163/8 767), totally with 8 904 live births.Conclusion:After more than 20 years of rapid development of PGD technology in China, the geographical accessibility has been significantly improved. PGD services are small and uneven. Comprehensive hospitals in the capital cities of the eastern and central regions are the main features of the technical configuration, and prefecture-level institutions and institutions in the western regions have also become important providers of PGD technology.

Objective:To analyze the configuration characteristics, service utilization trend and outcome of preimplantation genetic diagnosis (PGD) in China.Methods:A longitudinal study and a cross-sectional survey in 2020 were used to analyze the number of PGD facilities and the trend of service utilization in China from 2004 to 2020. The characteristics of 2020 technology configuration were described from different geographical distribution, city positioning, and organization categories. The number of couples treated, the number of egg retrieval cycles and the number of diagnostic cycles were used to evaluate the scale of the service.Results:From 2004 to 2020, the number of PGD institutions increased from only 4 (10.8%) in 2004 to 78 (19.0%) in 2020. Although the number of PGD cycles increased with it, the overall proportion was not high, ranging from 0.3% to 4.1%. A total of 21 241 PGD cycles were reported in 2020, accounting for 4.1% of the total number of egg retrievals. The PGD technology configuration in 2020 was mainly characterized by provincial capitals [82.1% (64/78)], public hospitals [94.9% (74/78)], and comprehensive hospitals [59.0% (46/78)]. There were significant differences in the median of the number of PGD egg retrieval cycles in eastern, central and western institutions, which were 203.0, 177.5 and 79.0, respectively. In 2020, the PGD clinical pregnancy rate was 60.2% (10 326/17 158), the delivery rate was 51.1% (8 767/17 158), the multiple delivery rate was 1.9% (163/8 767), totally with 8 904 live births.Conclusion:After more than 20 years of rapid development of PGD technology in China, the geographical accessibility has been significantly improved. PGD services are small and uneven. Comprehensive hospitals in the capital cities of the eastern and central regions are the main features of the technical configuration, and prefecture-level institutions and institutions in the western regions have also become important providers of PGD technology.