To investigate the infection status of two arboviruses,akabane orthobunyavirus(AKAV)and bluetongue virus(BTV),in cattle herds of Guizhou Province,we employed the indirect ELISA method to detect AKAV and BTV antibody levels in the present experiment.A total of 1504 bovine serum samples from 37 large-scale farms and 88 free-range households from 26 districts or coun-ties of 7 cities(prefectures)of Guizhou Province were collected to detect AKAV antibody levels.Additionally,1 241 serum samples from 30 large-scale farms and 15 free-range households in 19 districts or counties of 3 cities(prefectures)were tested for BTV antibody levels.Moreover,two influencing factors,breeding mode and sampling season,were statistically analyzed for their effects.The results showed that the overall positive rate of AKAV antibodies was 11.64％(175/1 504),with individual positive rates of 13.20％(123/934)and 9.12％(52/570)in large-scale farms and free-range households,respectively.No significant differences were observed between the two groups.However,the farm positive rate(64.86％,24/37)in large-scale farms was significantly higher than that(26.14％,23/88)in free-range households.Seasonal statistics showed that the positive rate was highest during the summer season at 60.00％(12/20).The total positive rate of BTV antibodies was 25.42％(222/1 241).The farm positive rate and individual positive rate in free-range households were 66.67％(10/15)and 41.91％(57/136),respectively.For large-scale farms,these rates were 60.00％(18/30)and 14.93％(165/1 105),respectively.The individual pos-itive rate in free-range households was significantly higher than that in large-scale farms.Seasonal statistics showed that the positive rates in summer and autumn seasons were 50.00％(5/10)and 72.41％(21/29),respectively,both of which were significantly higher than those in winter and spring seasons.All these findings indicated that both AKAV and BTV were present to a certain ex-tent in Guizhou Province,with seasonality.Furthermore,differences were observed between the different breeding modes.Our results could provide a data reference for the formulation of preven-tion and control measures for the two insect-borne diseases.

To establish a rapid visual detection method for Akabane virus(AKAV)on site,specific primers and probes based on the S fragment of AKAV were designed in this experiment.Corre-sponding groups were added to the primers or probes to fulfil the requirement of the combination of recombinase polymerase amplification(RPA)with lateral flow dipstick(LFD).The reaction temperature and time,concentrations of the primer and probe were optimized to establish the RPA-LFD method for detecting AKAV.After that,the specificity,sensitivity and clinical reliability of the method were evaluated.The results showed that after 20 minutes of reaction at 37 ℃,the test results could be read on LFD paper.There was no cross reaction against blue tongue virus,Pasteurella multocida,bovine infectious rhinotracheitis virus and bovine Mycoplasma bovis,and the detection limit was 2.5 × 100 copies/μL of standard plasmid.Detection of clinical samples showed a consistent results with that by RT-PCR method.These findings indicated that the RPA-LFD method established had the advantages of good specificity,high sensitivity,simple operation and visualization,and could be applied to clinical detection,which provides new technical support for the rapid diagnosis and prevention and control of AKAV.

To investigate the infection status of two arboviruses,akabane orthobunyavirus(AKAV)and bluetongue virus(BTV),in cattle herds of Guizhou Province,we employed the indirect ELISA method to detect AKAV and BTV antibody levels in the present experiment.A total of 1504 bovine serum samples from 37 large-scale farms and 88 free-range households from 26 districts or coun-ties of 7 cities(prefectures)of Guizhou Province were collected to detect AKAV antibody levels.Additionally,1 241 serum samples from 30 large-scale farms and 15 free-range households in 19 districts or counties of 3 cities(prefectures)were tested for BTV antibody levels.Moreover,two influencing factors,breeding mode and sampling season,were statistically analyzed for their effects.The results showed that the overall positive rate of AKAV antibodies was 11.64％(175/1 504),with individual positive rates of 13.20％(123/934)and 9.12％(52/570)in large-scale farms and free-range households,respectively.No significant differences were observed between the two groups.However,the farm positive rate(64.86％,24/37)in large-scale farms was significantly higher than that(26.14％,23/88)in free-range households.Seasonal statistics showed that the positive rate was highest during the summer season at 60.00％(12/20).The total positive rate of BTV antibodies was 25.42％(222/1 241).The farm positive rate and individual positive rate in free-range households were 66.67％(10/15)and 41.91％(57/136),respectively.For large-scale farms,these rates were 60.00％(18/30)and 14.93％(165/1 105),respectively.The individual pos-itive rate in free-range households was significantly higher than that in large-scale farms.Seasonal statistics showed that the positive rates in summer and autumn seasons were 50.00％(5/10)and 72.41％(21/29),respectively,both of which were significantly higher than those in winter and spring seasons.All these findings indicated that both AKAV and BTV were present to a certain ex-tent in Guizhou Province,with seasonality.Furthermore,differences were observed between the different breeding modes.Our results could provide a data reference for the formulation of preven-tion and control measures for the two insect-borne diseases.

To establish a rapid visual detection method for Akabane virus(AKAV)on site,specific primers and probes based on the S fragment of AKAV were designed in this experiment.Corre-sponding groups were added to the primers or probes to fulfil the requirement of the combination of recombinase polymerase amplification(RPA)with lateral flow dipstick(LFD).The reaction temperature and time,concentrations of the primer and probe were optimized to establish the RPA-LFD method for detecting AKAV.After that,the specificity,sensitivity and clinical reliability of the method were evaluated.The results showed that after 20 minutes of reaction at 37 ℃,the test results could be read on LFD paper.There was no cross reaction against blue tongue virus,Pasteurella multocida,bovine infectious rhinotracheitis virus and bovine Mycoplasma bovis,and the detection limit was 2.5 × 100 copies/μL of standard plasmid.Detection of clinical samples showed a consistent results with that by RT-PCR method.These findings indicated that the RPA-LFD method established had the advantages of good specificity,high sensitivity,simple operation and visualization,and could be applied to clinical detection,which provides new technical support for the rapid diagnosis and prevention and control of AKAV.

Objective To construct and validate a risk prediction model for poor inhalation in chronic obstructive pulmonary disease(COPD)patients receiving inhaler therapy,providing a decision support tool for personalized prevention of poor inhalation.Methods A cross-sectional study was conducted to collect data related to COPD patients receiving inhaler therapy,forming a dataset.The dataset was randomly divided into a training set and a test set in a ratio of 4∶1.Four different methods for missing value imputation,3 methods for variable feature selection,and 18 machine learning algorithms were employed to successfully construct 216 models on the training set.The monte carlo simulation method was used for resampling in the test set to validate the models,with the area under curve(AUC),accuracy,precision,recall,and F1 score used to evaluate model performance.The optimal model was selected to build the poor inhalation prediction platform.Results A study involving 308 patients with COPD found that 135(43.8%)were at risk of adverse inhalation.Using 33 predictor variables,216 risk prediction models were developed.Of these models,the ensemble learning algorithm yielded the highest average AUC of 0.844,with a standard deviation of 0.058[95%CI=(0.843,0.845)].The differences in predictive performance among the 216 models were statistically significant(P＜0.01).Under the ensemble learning algorithm,adherence to inhaler use(38.087 4%),inhaler satisfaction(25.680 1%),literacy(24.031 3%),number of inhalers(5.482 3%),age(4.204 5%)and number of acute exacerbations in the past year(2.184 7%)contributed most to the predictive model.The model exhibited superior performance,with an AUC of 0.869 3,an accuracy of 83.87%,a precision of 86.96%,a recall of 74.07%,and an F1 score of 0.8.Conclusion This study has developed a predictive model for poor inhalation risk in COPD inhaler therapy patients using machine learning algorithms,which exhibits strong predictive capabilities and holds potential clinical application value.

ObjectiveTo explore the distribution characteristics of traditional Chinese medicine （TCM） syndrome elements of macroangiopathy in patients with type 2 diabetes mellitus （T2DM） and the key elements of occurrence， development and progression of disease. MethodsA multicenter cross-sectional study was conducted to enroll 445 T2DM patients from five hospitals， and according to the presence or absence of macroangiopathy， the patients were divided into a T2DM group （120 cases） and a diabetic macroangiopathy （DM） group （325 cases）. Patients in DM group were divided into grade Ⅰ， Ⅱ， Ⅲ and Ⅳ according to the peripheral vascular color Doppler ultrasound results and the vascular anomalies classification standard. The general data including gender， age， duration of T2DM and body mass index （BMI） were collected， and the data of four examinations were obtained for syndrome differentiation. According to the diagnostic criteria of TCM syndrome elements， the patients can be divided into 9 patterns including qi deficiency， blood deficiency， yin deficiency， yang deficiency， qi stagnation， blood stasis， excess heat， and excess cold. The general data and distribution of TCM syndrome elements were compared between the two groups. The distribution of TCM syndrome elements in different vascular anomalies grades in the DM group was analyzed. Logistic regression analysis was used to explore the influence of various TCM syndrome elements on the occurrence of macroangiopathy in T2DM. ResultsThere was no significant difference in gender and BMI between groups （P＞0.05）. The age and duration of diabetes in the DM group were older and longer than those in the T2DM group （P＜0.01）. With the increase of age and prolonged course of disease， the severity of diabetic macroangiopathy increases gradually （P＜0.05 or P＜0.01）. There was no significant difference in BMI and course of disease among the different TCM syndrome elements （P＞0.05）. The average age of patients with blood stasis syndrome was the oldest （P＜0.05）. There was significant difference in gender distribution between the excess heat syndrome and yin deficiency syndrome （P＜0.05）. A total of 240 TCM syndrome elements were extracted from the T2DM group， while 731 TCM syndrome elements extracted from the DM group. The top two high-frequency syndrome elements in the two groups were qi deficiency and yin deficiency， with a frequency of larger than 50%. The distribution of phlegm-damp syndrome and blood-stasis syndrome were significantly higher in the DM group than in the T2DM group （P＜0.01）. There were significant differences in the distribution of qi deficiency syndrome， yin deficiency syndrome， phlegm-damp syndrome， blood stasis syndrome， and excess heat syndrome among different grades of vascular anomalies （P＜0.01）； qi deficiency and yin deficiency were both high-frequency TCM syndrome elements in patients at grades 0 to Ⅲ； phlegm-damp syndrome increased in frequency with the progression of the disease from grades 0 to Ⅳ， and the frequency of blood stasis syndrome showed an overall upward trend. The frequency of phlegm-dampness syndrome increased from grades 0 to Ⅳ with the progression of the disease， and the frequency of blood stasis syndrome showed an overall upward trend. Logistic regression analysis showed that phlegm-damp syndrome and blood stasis syndrome were important TCM syndrome elements related to the vascular anomalies degree of macrovascular disease in T2DM （P＜0.05 or P＜0.01）. ConclusionQi deficiency and yin deficiency are the basic TCM syndrome elements throughout the whole process of T2DM and diabetic macrovascular disease. Phlegm-damp and blood stasis are related to the degree of vascular anomalies in diabetic macrovascular disease and are the key TCM syndrome elements in the progression of macroangiopathy in T2DM.

Objective:To evaluate the value of contrast-enhanced ultrasound parameters, systemic immune-inflammation index (SII) and neutrophil to lymphocyte ratio (NLR) in the prognosis of prostate cancer after radical resection.Methods:The clinical data of 94 patients with radical resection of prostate cancer admitted to the Ankang Hospital of Traditional Chinese Medicine from February 2018 to February 2023 were retrospectively analyzed. The patients were followed up for 2 years after surgery. According to the follow-up results, the patients were divided into a recurrence group (37 cases) and a non-recurrence group (57 cases). Basic data [including age, body mass index (BMI), diabetes, hypertension, clinical stage, course of disease, Gleason score, surgical method, and American Society of Anesthesiologists (ASA) grade], preoperative SII and NLR, and postoperative 1 d contrast-enhanced ultrasound data of the two groups were collected, and compared between groups. The risk factors of postoperative recurrence in patients with prostate cancer were screened by unconditional logistic step-by-step regression analysis. The effectiveness of contrast ultrasound parameters, SII and NLR in evaluating postoperative recurrence of prostate cancer was analyzed by receiver operating characteristic (ROC) curve. At the same time, the prediction model of postoperative recurrence of prostate cancer patients was constructed and verified by ROC curve.Results:There were no significant differences in age, BMI, diabetes, hypertension, course of disease, surgical method and ASA classification between the two groups (all P>0.05). The proportion of clinical stage Ⅲ to Ⅳ, Gleason score, peak intensity (PI), mean time of passage (MTT), area under the curve (AUC), half time of intensity decline (HT), ascending branch slope (WIS), platelet count, SII and NLR in the non-recurrent group were lower than those in the recurrent group, and the differences were statistically significant (all P<0.05). Multivariate logistic regression analysis confirmed that clinical stage Ⅲ to Ⅳ, Gleason score, PI, MTT, AUC, HT, WIS, platelet count, SII and NLR were risk factors for postoperative recurrence of prostate cancer patients (all P<0.05). ROC curve analysis results showed that the areas under the curve of PI, MTT, AUC, HT, WIS, SII and NLR for postoperative recurrence of prostate cancer patients were 0.850, 0.923, 0.867, 0.856, 0.788, 0.746 and 0.932, respectively. The sensitivity was 0.811, 0.838, 0.784, 0.811, 0.703, 0.676, 0.892, respectively. According to the results of multivariate logistic regression analysis, the prediction model of postoperative recurrence of prostate cancer patients was fitted as Logit( P)=-3.661+ 0.464×clinical stage+ 0.611×Gleason score+ 0.617×PI+ 0.901×MTT+ 1.071×AUC+ 0.952×HT+ 0. 728×WIS+ 0.568×platelet count+ 0.628×SII+ 0.750×NLR. ROC curve results showed that the area under the curve of the model for predicting postoperative recurrence of prostate cancer patients was 0.953, standard error was 0.035, 95% CI was 0.884-1.000, sensitivity was 0.958, and specificity was 0.911. Conclusions:Contrast-enhanced ultrasound parameters, SII and NLR have certain value in the prognosis evaluation of prostate cancer after radical resection, and can be used as clinical indicators to evaluate whether the recurrence is possible.

Objective:To evaluate the value of contrast-enhanced ultrasound parameters, systemic immune-inflammation index (SII) and neutrophil to lymphocyte ratio (NLR) in the prognosis of prostate cancer after radical resection.Methods:The clinical data of 94 patients with radical resection of prostate cancer admitted to the Ankang Hospital of Traditional Chinese Medicine from February 2018 to February 2023 were retrospectively analyzed. The patients were followed up for 2 years after surgery. According to the follow-up results, the patients were divided into a recurrence group (37 cases) and a non-recurrence group (57 cases). Basic data [including age, body mass index (BMI), diabetes, hypertension, clinical stage, course of disease, Gleason score, surgical method, and American Society of Anesthesiologists (ASA) grade], preoperative SII and NLR, and postoperative 1 d contrast-enhanced ultrasound data of the two groups were collected, and compared between groups. The risk factors of postoperative recurrence in patients with prostate cancer were screened by unconditional logistic step-by-step regression analysis. The effectiveness of contrast ultrasound parameters, SII and NLR in evaluating postoperative recurrence of prostate cancer was analyzed by receiver operating characteristic (ROC) curve. At the same time, the prediction model of postoperative recurrence of prostate cancer patients was constructed and verified by ROC curve.Results:There were no significant differences in age, BMI, diabetes, hypertension, course of disease, surgical method and ASA classification between the two groups (all P>0.05). The proportion of clinical stage Ⅲ to Ⅳ, Gleason score, peak intensity (PI), mean time of passage (MTT), area under the curve (AUC), half time of intensity decline (HT), ascending branch slope (WIS), platelet count, SII and NLR in the non-recurrent group were lower than those in the recurrent group, and the differences were statistically significant (all P<0.05). Multivariate logistic regression analysis confirmed that clinical stage Ⅲ to Ⅳ, Gleason score, PI, MTT, AUC, HT, WIS, platelet count, SII and NLR were risk factors for postoperative recurrence of prostate cancer patients (all P<0.05). ROC curve analysis results showed that the areas under the curve of PI, MTT, AUC, HT, WIS, SII and NLR for postoperative recurrence of prostate cancer patients were 0.850, 0.923, 0.867, 0.856, 0.788, 0.746 and 0.932, respectively. The sensitivity was 0.811, 0.838, 0.784, 0.811, 0.703, 0.676, 0.892, respectively. According to the results of multivariate logistic regression analysis, the prediction model of postoperative recurrence of prostate cancer patients was fitted as Logit( P)=-3.661+ 0.464×clinical stage+ 0.611×Gleason score+ 0.617×PI+ 0.901×MTT+ 1.071×AUC+ 0.952×HT+ 0. 728×WIS+ 0.568×platelet count+ 0.628×SII+ 0.750×NLR. ROC curve results showed that the area under the curve of the model for predicting postoperative recurrence of prostate cancer patients was 0.953, standard error was 0.035, 95% CI was 0.884-1.000, sensitivity was 0.958, and specificity was 0.911. Conclusions:Contrast-enhanced ultrasound parameters, SII and NLR have certain value in the prognosis evaluation of prostate cancer after radical resection, and can be used as clinical indicators to evaluate whether the recurrence is possible.

AIM: To investigate the mechanism of baicalin-induced apoptosis in human laryngeal cancer cells. METHODS: AMC-HN-8 cells were selected for the study, and baicalin was applied to the cells at different concentrations (0, 10, 30, 100, and 300 μmol/L), and the half-inhibitory concentration (IC50) was measured by the CCK-8 method. Bax, cleaved-caspase-3, Cyto-c, IRF4 protein expression by protein blotting (Western blot); miR-125b-5p and IRF4 expression by RT-qPCR. Dual-luciferase reporter gene validation of Targetscan prediction (binding of miR-125b-5p to IRF4-3'UTR); apoptosis and necrosis inhibitors explore the way baicalein induces death in laryngeal cancer cells. AMC-HN-8 was then divided into blank group, baicalein (IC50), miR-125b-5p inhibitor group, baicalein + inhibitor NC group, baicalein+miR-125b-5p inhibitor group, and cell invasion and clone formation assays to detect cell invasion and proliferation ability, respectively. Apoptosis was detected by flow cytometry. RESULTS: Baicalein inhibited the proliferation of AMC-HN-8 cells in a dose-dependent manner with an IC50 value of 47.31 μmol/L. Compared with the blank group, 47.31 μmol/L baicalin induced apoptosis and inhibited cell invasion, while upregulating the expression of miR-125b-5p and suppressing the mRNA and protein levels of IRF4. The luciferase results showed that the miR-125b-5p mimic was able to inhibit the activity of the IRF4-3'UTR promoter relative to the NC mimic (mimic) group. Baicalein induces laryngeal cancer cell death in an apoptotic manner. In addition, the combination of 47.31 μmol/L baicalin and miR-125b-5p inhibitor affected the behavior of AMC-HN-8 cells, showing that compared with the blank group, the baicalin group showed a decrease in the number of cell clones, weakened invasion ability, and increased apoptosis; the miR - 125b-5p inhibitor group showed an increase in the number of cell clones, enhanced invasion ability and decreased apoptosis. The baicalin+ inhibitor NC group was consistent with baicalin, with no significant effect of inhibitor NC on cell behavior. The cloning, invasion, and apoptosis of cells in the baicalin+miR-125b-5p inhibitor group were intermediate between the baicalin and miR-125b-5p inhibitor groups. CONCLUSION: Baicalin inhibits the proliferation of AMC-HN-8 cells, and the mechanism may be related to miR-125b-5p targeting to inhibit the expression of IRF4, inducing the pro-apoptotic proteins Bax, cleaved-caspase3, and Cyto-c, and inhibiting the apoptosis suppressor protein Bcl-2 thereby inducing apoptosis.