The present study aimed to establish an LC-MS/MS method for the quantification of tiamulin in minipig plasma and to further conduct a pharmacokinetic comparison of different formulations. The plasma samples were extracted with acetonitrile (meloxicam as internal standard), separated on a C18 column, and quantified by multiple reaction monitoring mode (ESI+). Sanyuan minipigs were used as experimental animals. Plasma samples were collected after intravenous injection (10 mg/kg) and intragastric administration (20 mg/kg). The method showed good linearity, with intra- and inter-batch RSD of 1.00%–8.13% and RE within ±15%. The extraction recovery, matrix effect and stability of the analytical methods met the relevant requirements. Tiamulin fumarate active pharmaceutical ingredient was intravenously administered, with c0 of about (4383.73±2676.78) ng/mL, AUC0-t of about (4803.50±965.68) h·ng/mL, t1/2 of about (4.66±1.68) h, and CL of about (2.14±0.46) L/(kg·h). Three tiamulin formulations were intragastrically administered, with cmax of (552.00±328.55), (545.00±136.97) and (590.60±237.02) ng/mL, tmax of (1.47±0.68), (0.69±0.75) and (0.72±0.72) h, and F of 24.85%, 15.28% and 16.97%, respectively. The validated method meets the requirements for biological sample analysis and is applicable for the pharmacokinetic evaluation of tiamulin formulations in minipigs.

Objective To investigate the correlation between arterial stiffness(AS)and incident chronic kidney disease(CKD)among the elderly individuals taking health checkup.Methods A retrospective study was conducted on 857 elderly individuals without CKD at baseline who taking physical exams in our medical center from December 2009 to May 2021.Their clinical and labora-tory data were collected.Brachial-ankle pulse wave velocity(baPWV)was used to assess the se-verity of AS,and then the subjects were divided into normal elasticity group(201 cases),and moderate(490 cases)and severe AS group(166 cases).Kaplan-Meier curves were plotted to dis-play cumulative incidence rates of incident CKD across different AS groups.Restricted cubic splines(RCS)and Cox regression models were applied to analyze the correlation of baPWV and incident CKD risk.Results The severe AS group had significantly advanced age,greater ratio of hypertension,larger waist circumference,higher HR,SBP and DBP,increased urinary albumin/creatinine ratio(UACR),elevated levels of TG and fasting blood glucose,and baPWV than the normal elasticity group(P＜0.05).During the follow-up period,37 participants developed CKD.The incidence of CKD was obviously higher in the severe AS group than the normal arterial elas-ticity group(9.04％vs 3.48％).RCS analysis revealed a U-shaped relationship between baPWV and incident CKD risk.When baPWV ≥1 400 cm/s,each standard deviation increase in baPWV indicates the risk of incident CKD increasing by 71％(HR=1.71,95％CI:1.30-2.25,P＜0.01).Regardless of adjustment for covariates or not,baPWV remained positive correlation with inci-dent CKD risk(P＜0.05).Conclusion Among the elderly individuals undergoing health check-up,increased AS severity is significantly associated with higher risk of incident CKD when baP-WV ≥1400 cm/s.

Objective To investigate the correlation between arterial stiffness(AS)and incident chronic kidney disease(CKD)among the elderly individuals taking health checkup.Methods A retrospective study was conducted on 857 elderly individuals without CKD at baseline who taking physical exams in our medical center from December 2009 to May 2021.Their clinical and labora-tory data were collected.Brachial-ankle pulse wave velocity(baPWV)was used to assess the se-verity of AS,and then the subjects were divided into normal elasticity group(201 cases),and moderate(490 cases)and severe AS group(166 cases).Kaplan-Meier curves were plotted to dis-play cumulative incidence rates of incident CKD across different AS groups.Restricted cubic splines(RCS)and Cox regression models were applied to analyze the correlation of baPWV and incident CKD risk.Results The severe AS group had significantly advanced age,greater ratio of hypertension,larger waist circumference,higher HR,SBP and DBP,increased urinary albumin/creatinine ratio(UACR),elevated levels of TG and fasting blood glucose,and baPWV than the normal elasticity group(P＜0.05).During the follow-up period,37 participants developed CKD.The incidence of CKD was obviously higher in the severe AS group than the normal arterial elas-ticity group(9.04％vs 3.48％).RCS analysis revealed a U-shaped relationship between baPWV and incident CKD risk.When baPWV ≥1 400 cm/s,each standard deviation increase in baPWV indicates the risk of incident CKD increasing by 71％(HR=1.71,95％CI:1.30-2.25,P＜0.01).Regardless of adjustment for covariates or not,baPWV remained positive correlation with inci-dent CKD risk(P＜0.05).Conclusion Among the elderly individuals undergoing health check-up,increased AS severity is significantly associated with higher risk of incident CKD when baP-WV ≥1400 cm/s.

Objective To analyze the clinical characteristics of lung adenocarcinoma patients with positive EGFR mutations detected in pleural effusion.Methods We retrospectively analyzed the clinical characteristics including gender,age,smoking history,presence of other underlying diseases(such as COPD,cardiovascular disease,and diabetes),site of pleural fluid,feature of pleural fluid,and TNM stage in patients with lung adenocar-cinoma who had been admitted to the first Affiliated Hospital of Bengbu Medical College from 2020.01 to 2022.12 for the first time by the detection of EGFR mutation positive in pleural effusion.The data were statistically analyzed using the SPSS 26.0 software.Results A total of 126 patients were screened for enrollment,including 61 patients(48.41%)with EGFR exon 19 deletion mutation(19del),56 patients(44.44%)with exon 21 L858R mutation(21L858R),and 9 patients(7.14%)with non-classical mutations.Univariate analysis showed that the three muta-tion subtypes were statistically significant in terms of gender,age,smoking history,and presence of COPD(P<0.05 for all comparisons),but not in terms of pleural fluid site,feature of pleural fluid,tumor size,and presence of cardiovascular disease,diabetes mellitus,presence of distant metastases,and mediastinal lymph node metastases(P>0.05 for all comparisons);Multivariate analysis showed that 21 L858R mutation was more likely to be found in male,older age,non-smoking,and presence of COPD than 19del mutation;non-classical mutation was more likely to be found in male than 19del mutation.Conclusions There are significant differences among the three mutation subtypes in sex,age,smoking history,and presence of COPD,but not in pleural fluid location,feature of pleural fluid,tumor size,presence of cardiovascular disease or diabetes mellitus,presence of distant metastases,or medias-tinal lymph node metastases;Among lung adenocarcinoma patients with positive EGFR mutations in pleural fluid,21 L858R mutation mostly occurs in male,older age,non-smokers,and those complicated with COPD,while non-classical mutation mainly develops in male.However,more case studies are needed to confirm the above conclusions.

Background::In vitro fertilization (IVF) has emerged as a transformative solution for infertility. However, achieving favorable live-birth outcomes remains challenging. Current clinical IVF practices in IVF involve the collection of heterogeneous embryo data through diverse methods, including static images and temporal videos. However, traditional embryo selection methods, primarily reliant on visual inspection of morphology, exhibit variability and are contingent on the experience of practitioners. Therefore, an automated system that can evaluate heterogeneous embryo data to predict the final outcomes of live births is highly desirable. Methods::We employed artificial intelligence (AI) for embryo morphological grading, blastocyst embryo selection, aneuploidy prediction, and final live-birth outcome prediction. We developed and validated the AI models using multitask learning for embryo morphological assessment, including pronucleus type on day 1 and the number of blastomeres, asymmetry, and fragmentation of blastomeres on day 3, using 19,201 embryo photographs from 8271 patients. A neural network was trained on embryo and clinical metadata to identify good-quality embryos for implantation on day 3 or day 5, and predict live-birth outcomes. Additionally, a 3D convolutional neural network was trained on 418 time-lapse videos of preimplantation genetic testing (PGT)-based ploidy outcomes for the prediction of aneuploidy and consequent live-birth outcomes.Results::These two approaches enabled us to automatically assess the implantation potential. By combining embryo and maternal metrics in an ensemble AI model, we evaluated live-birth outcomes in a prospective cohort that achieved higher accuracy than experienced embryologists (46.1% vs. 30.7% on day 3, 55.0% vs. 40.7% on day 5). Our results demonstrate the potential for AI-based selection of embryos based on characteristics beyond the observational abilities of human clinicians (area under the curve: 0.769, 95% confidence interval: 0.709–0.820). These findings could potentially provide a noninvasive, high-throughput, and low-cost screening tool to facilitate embryo selection and achieve better outcomes. Conclusions::Our study underscores the AI model’s ability to provide interpretable evidence for clinicians in assisted reproduction, highlighting its potential as a noninvasive, efficient, and cost-effective tool for improved embryo selection and enhanced IVF outcomes. The convergence of cutting-edge technology and reproductive medicine has opened new avenues for addressing infertility challenges and optimizing IVF success rates.

Objective:To investigate the effects of hydroxysafflor yellow A (HSYA) on depressive-like behavior and expression of type A γ-aminobutyric acid receptor(GABAAR)in hippocampus of chronic restraint stress model mice.Methods:The SPF grade male C57BL/6C mice were divided into Control group, HSYA group, Model group, Model + HSYA group and Model + fluoxetine group according to random number table method, with 12 mice in each group.Mice model of depression was established by chronic restraint stress.Mice in HSYA group and Model+ HSYA group were intraperitoneally injected with HSYA(20 mg/kg), mice in Model+ fluoxetine group were injected intraperitoneally with fluoxetine (10 mg/kg), and mice in Control group and Model group administered with 0.9% sodium chloride solution intraperitoneally once a day for 14 days.Then, the forced swimming test (FST) and tail suspension test (TST) were performed to evaluate the depressive-like behavior of mice, and the protein expression levels of different subtypes of GABAAR in the hippocampus of mice were determined by Western blot.SPSS 19.0 and GraphPad Prism 8.0 software were used for data statistical analysis and mapping.One-way ANOVA was used for comparison among groups, and Tukey-HSD test was used for further pairwise comparison.Results:(1) In the behavioral tests, there were significant differences in swimming immobility time of FST and tail suspension immobility time of TST among the five groups ( F=21.59, 20.81, both P<0.05). The swimming immobility time ((143.91±9.97) s) and tail suspension immobility time (( 107.00±6.54) s) in Model group were higher than those in Control group ((52.92±6.70) s, ( 43.50±5.96) s, both P<0.05). There were no significant difference in swimming immobility time and tail suspension immobility time between Model+ HSYA group ((26.17±7.69)s, ( 20.17±7.89)s) and Model+ fluoxetine group ((61.60±16.22)s, (34.14±10.74)s)(both P>0.05), but the swimming immobility time and tail suspension immobility time in these two groups were lower than those in Model group (both P<0.05). (2) The Western blot results showed that there were significant differences in the expression of GABAARβ1 and GABAARβ2 protein in hippocampus among the four groups ( F=12.21, 11.40, both P<0.05). The expression levels of GABAARβ1(45.60±10.76) and GABAARβ2 (46.27±4.82) protein in hippocampus of Model group were lower than those in Control group ((100.00±3.44), (100.00±3.26), both P<0.05). Compared to Model group, the expression of GABAARβ1 (79.91±5.00) and GABAARβ2 (79.08±5.53) protein in hippocampus of Model+ HSYA group were higher (both P<0.05). In addition, the expression of GABAARα1 and GABAARγ1 proteins in hippocampus were not significantly different among the four groups( F=0.23, 0.10, both P>0.05). Conclusion:HSYA can effectively alleviate depressive-like behavior in depression model mice, which may be related with the upregulation of GABAARβ1 and GABAARβ2 of hippocampus tissue.

Prediction of the interactions between small molecules and their targets play important roles in various applications of drug development, such as lead discovery, drug repurposing and elucidation of potential drug side effects. Therefore, a variety of machine learning-based models have been developed to predict these interactions. In this study, a model called auxiliary multi-task graph isomorphism network with uncertainty weighting (AMGU) was developed to predict the inhibitory activities of small molecules against 204 different kinases based on the multi-task Graph Isomorphism Network (MT-GIN) with the auxiliary learning and uncertainty weighting strategy. The calculation results illustrate that the AMGU model outperformed the descriptor-based models and state-of-the-art graph neural networks (GNN) models on the internal test set. Furthermore, it also exhibited much better performance on two external test sets, suggesting that the AMGU model has enhanced generalizability due to its great transfer learning capacity. Then, a naïve model-agnostic interpretable method for GNN called edges masking was devised to explain the underlying predictive mechanisms, and the consistency of the interpretability results for 5 typical epidermal growth factor receptor (EGFR) inhibitors with their structure‒activity relationships could be observed. Finally, a free online web server called KIP was developed to predict the kinome-wide polypharmacology effects of small molecules (http://cadd.zju.edu.cn/kip).

【Objective】 To explore the clinical efficacy of traditional Chinese medicine(TCM) therapy in patients with transfusion related iron overload diagnosed with chronic aplastic anemia (CAA). 【Methods】 A total of 115 patients with CAA and iron overload who had been admitted to Zhejiang Provincal Hospital of TCM from February 2015 to December 2016 were studied. They were assigned to treatment group(n=69), positive control group(n=16), and negative control group(n=30) according to different treatment plan designed in advance. Patients in the treatment group were treated with TCM of "Bushen Huoxue" recipe once a day, with the formula mixed with astragalus 40 g, rehmannia 12 g, cornus 12 g, deerhorn glue melting by heat 12 g, atractylodes 20 g, radix paeoniae alba 20 g, curculigo orchioides 10 g, herba epimedii 10 g, cistanche 12 g, cassia stem 10 g, dried orange peel 8 g, poria 10, gingembre 6 g, angelica sinensis 20 g, salvia miltiorrhiza 20 g, zedoary 9 g, leonurus 30 g, gromwell 15 g, and prepared liquorice root 6 g. Patients in positive control group were treated with desferrioxamine for more than 8 h per day, 5 to 7 days per week. Patients in negative control group were treated with basic treatment.The serum ferritin (SF) and cytokines of patients of the three groups before and 3 months after therapy were detected according to the blood-stasis of TCM symptom rating scale, and the correlation between the decrease of ferritin after treatment and the improvement of blood stasis syndrome score was analyzed. 【Results】 The level of SF of the treatment group, positive and negative control group before treatment were 1 881.63±1 386.81 vs 6 581.36±5 180.96 vs1 974.25±1 753.06, and were 2 040.14±1 484.27 vs 4 169.18±3 631.64(P<0.05)vs 2 699.80±2 352.34(P<0.05) 3 months after treatment. The treatment was effctive in 16 patients in treatment group, accounted for 23.19%(16/69). The score of blood stasis syndrome of the three groups before and after treatment were 4.26±1.45vs 6.88±1.31 vs 4.17±1.18 and 4.42±1.43 vs 5.00±0.89 vs 4.67±1.51(P<0.01), respectively. The effective rate of improving blood stasis syndrome score in the treatment group was up to 26.09%. The decrease of serum ferritin was positively correlated with the improvement of blood stasis score (P < 0.01). The levels of IL-6 and IL-10 in treatment group were 20.79±14.14 and 56.27±25.54 before treatment, 13.00±6.48 and 41.02±9.93(P<0.05)3 months after treatment, respectively. 【Conclusion】 "Bushen Huoxue" therapy can stabilize the the level of SF and improve the blood stasis syndrome in CAA patients with iron overload.

Objective: To investigate the biological characteristics of high hyperdiploid and triploid/tetraploid acute myeloid leukemia (HH/TT-AML) and its relationship with prognosis. Methods: The clinical data of 28 patients with newly diagnosed HH/TT-AML during March 2006 and June 2017 were retrospectively analyzed, and the factors influencing prognosis were analyzed by the Kaplan-Meier method. Results: The karyotypes of HH/TT-AML patients were mainly 49 chromosomes, accounting for 39.3% (11/28), followed by 50-55 chromosomes, accounting for 32.1% (9/28). The karyotypes of high hyperdiploid acute myeloid leukemia (HH-AML) patients were more likely to be +8 (77.3%, 17/22) or +21 (54.5%, 12/22). The survival analysis showed that the overall survival rate of HH/TT-AML patients with -5/5q-, -7/7q-, -17/der(17p) or der(3q) was significantly lower than that without these abnormalities (4.1 months vs 10.1 months,P＜0.05). There was no significant difference in the overall survival rate between triploid/tetraploid acute myeloid leukemia (TT-AML) patients and HH-AML patients (8.4 months vs 7.2 months,P＞0.05). The overall survival rate of the patients with allogeneic hematopoietic stem cell transplantation was significantly longer than that with chemotherapy alone (25.4 months vs 4.1 months,P＜0.01). Conclusion HH/TT AML patients are highly heterogeneous. The identification of poor prognosis-related chromosome abnormalities is helpful for the stratification of prognosis. The overall survival time of these patients is short. Early allogeneic hematopoietic stem cell transplantation after remission by combination chemotherapy can significantly improve the prognosis of these patients.

Objective To investigate the clinical characteristics of metabolically healthy obese ( MHO) individuals, and to explore the risk of progression into metabolic disorders after 3 years. Methods A total of 3943 residents in Jining City were evaluated twice from February 2012 to August 2015, and 3766 individuals were enrolled excluding those with missing data. Of the subjects, 875 subjects were screened as metabolic normal population, which were divided into MHO(n = 127), metabolically healthy overweight (MHOW, n = 386), and metabolically healthy normal weight ( MHNW, n = 362) groups. T test, x2 test, and logistic regression analysis were used for data analysis. Results The incidence of MHO was 11. 63％ (127 / 1092) in obesity, and the proportion of MHO in females was higher than that in males(13. 91％ vs 7. 82％ , P<0. 05). Compared with MHNW group, the levels of HbA1C , fasting insulin, low density lipoprotein-cholesterol ( LDL-C), triglyceride ( TG), glutamyl transpeptidase (GGT), systolic blood pressure(SBP), diastolic blood pressure(DBP), and waist circumference(WC) were higher in MHO while glomerular filtration rate (GFR) and high density lipoprotein-cholesterol (HDL-C) were lower(all P<0. 05); and fasting insulin, LDL-C, TG, GGT, SBP, WC were higher in MHOW while HDL-C was lower (all P<0. 05). The levels of fasting insulin, TG, SBP, WC were higher in MHO while GFR and HDL-C were lower compared with MHOW(all P<0. 05). Following up for 3 years, the incidences of dyslipidemia in MHNW, MHOW, and MHO were 17. 96％ (65 / 362), 32. 90％ (127 / 386), 42. 52％ (54 / 127), respectively, with significant difference among three groups(P<0. 05). The incidences of hyperglycemia in the three groups were 20. 17％ (73 / 362), 22. 80％(88 / 386), 26. 77％ (34 / 127), respectively, without significant difference among groups ( all P > 0. 05). After adjustment for some factors including sex, age, fasting insulin, glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, GGT, and creatinine, the risks of dyslipidemia in MHO ( OR = 2. 193, 95％ CI 1. 359-3. 539, P<0. 05) and MHOW(OR= 1. 705, 95％ CI 1. 190-2. 443, P<0. 05) were significantly increased as compared with MHNW. Conclusion Compared with MHNW individuals, MHOW/ MHO individuals show an obviously different clinical feature as well as with higher risks of dyslipidemia after 3 years.