Ovulatory disorders are mainly characterized by abnormal follicular maturation or ovulation， with complex etiologies and a lack of effective prevention and treatment methods. Autophagy dysfunction is closely related to the generation and progression of ovulatory disorders. This article systematically elucidates the mechanisms of TCM on follicular development and ovulatory disorders from the perspective of autophagy. It also reviews relevant studies on how TCM regulates autophagy to influence follicular development and improve ovulatory disorders. The findings reveal that TCM monomers/active ingredients （leonurine， total flavonoids from Eucommia ulmoides， alpinetin， icariin， etc.） and compound formulas （including Cangfu daotan decoction， Guishen yugong decoction， Zhuluan decoction， Yishen yangluan formula， Guishen pill， etc.） improve the follicular microenvironment， regulate sex hormone levels， and reduce follicular atresia by regulating autophagy-related genes and signaling pathways such as phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin and AMP-activated protein kinase. These actions thereby promote normal follicular development and ovulation， and delay ovarian aging. Most research in this field is based on cellular and animal experiments， often focusing on a single signaling pathway or factor. Some studies fail to fully reflect the individualized treatment characteristics of TCM that emphasize “syndrome differentiation and treatment”， highlighting the urgent need for further investigation.

This study aims to establish BHK-21 stable cell lines expressing APN from four species(human,pig,dog,and cat),the APN fragments were amplified from pEGFP-C1-APN plasmids of the four species stored in the laboratory to generate the recombinant plasmids pcDNA4.0-APN.Af-ter the recombinant plasmids were transfected into BHK-21 cells,the stable BHK-21 cell lines ex-pressing the APNs were selected by two rounds of limited dilution.The constructed BHK-21 cell lines were identified by indirect immunofluorescence assay(IFA),and their susceptibility to PD-CoV and TGEV was tested for these four cell lines.Virus infection experiments revealed that PD-CoV infected cells expressing human,pig,and dog APNs,while it did not infect cells expressing cat APN.Simultaneously,TGEV infected cells expressing pig,dog,and cat APNs,but did not infect cells expressing human APN.The results suggest that the risk of cross-species infection for different coronaviruses and the established cell line can be used effectively to evaluate the virus in-fection.The findings also revealed that PDCoV has the potential risk of cross-species infection of human and dog,and TGEV has the potential risk of cross-species infection of dog and cat.These results provide a basis for the prevention and control strategy of coronaviruses.

This study aims to establish BHK-21 stable cell lines expressing APN from four species(human,pig,dog,and cat),the APN fragments were amplified from pEGFP-C1-APN plasmids of the four species stored in the laboratory to generate the recombinant plasmids pcDNA4.0-APN.Af-ter the recombinant plasmids were transfected into BHK-21 cells,the stable BHK-21 cell lines ex-pressing the APNs were selected by two rounds of limited dilution.The constructed BHK-21 cell lines were identified by indirect immunofluorescence assay(IFA),and their susceptibility to PD-CoV and TGEV was tested for these four cell lines.Virus infection experiments revealed that PD-CoV infected cells expressing human,pig,and dog APNs,while it did not infect cells expressing cat APN.Simultaneously,TGEV infected cells expressing pig,dog,and cat APNs,but did not infect cells expressing human APN.The results suggest that the risk of cross-species infection for different coronaviruses and the established cell line can be used effectively to evaluate the virus in-fection.The findings also revealed that PDCoV has the potential risk of cross-species infection of human and dog,and TGEV has the potential risk of cross-species infection of dog and cat.These results provide a basis for the prevention and control strategy of coronaviruses.

OBJECTIVE To evaluate the quality of diagnosis and treatment guidelines and expert consensuses on childhood immune thrombocytopenic purpura(ITP)published domestically and internationally,in order to provide reference for clinical practice and future guideline/expert consensus development and improvement.METHODS A systematic search was conducted across multiple databases,including PubMed,Cochrane Library,Embase,CNKI,Wanfang data,VIP,CBM;additionally,supplementary searches were carried out on websites such as Medlive,the Chinese Medical Association's official website,and National Institute for Health and Clinical Excellence in the UK.The retrieval time ranged from the inception to September 2,2024.Researchers who had undergone systematic training independently evaluated the methodology and report quality included in the guideline/consensus using the Appraisal of Guidelines Research and Evaluation Ⅱ(AGREE Ⅱ)and the Reporting Items for Practice Guidelines in Healthcare(RIGHT).RESULTS A total of 11 guidelines/consensuses were included.The average scores for the six domains of AGREE Ⅱ tool respectively were"range and purpose"[(66.67±17.98)%],"participants"[58.33%(13.89%,73.61%)],"rigor"[(41.81±23.85)%],"clarity"[(69.57±19.35)%],"applicability"[(35.98±17.83)%],and"independence"[27.08%(0,75.00%)];out of 11 articles,9 had a recommendation level of B,2 had a recommendation level of C,and there were no A-level articles.The average reporting rates of the 7 areas in the RIGHT tool were"basic information"[(72.35±12.95)%],"background"[(54.55±15.40)%],"evidence"[(36.36±24.81)%],"recommended opinions"[(53.25±19.20)%],"review and quality assurance"[0(0,25.00%)],"funding and conflict of interest statement and management"[12.50%(0,25.00%)],and other aspects[8.33%(0,50.00%)].In addition,there was no statistically significant difference in the AGREE Ⅱ and RIGHT scores between the guidelines and consensuses(P＞0.05).CONCLUSIONS The overall quality of the guidelines and consensuses included in this study is not high,with a recommended level of B or C.It is recommended that clinical decision-making prioritize referring to the relatively high-quality guideline/consensus among them.The quality of evidence in the existing traditional Chinese medicine guidelines for children with ITP needs to be improved,and there is no integrated guideline/consensus for traditional Chinese and Western medicine.It is recommended to revise or write relevant guideline/consensus according to the requirements of AGREE Ⅱ and RIGHT in various fields to guide clinical practice.

Objective To investigate the effect of circular RNA_003213 (hsa_circ_003213) targeting microRNA-145-5p (miR-145-5p) on the proliferation, migration and invasion of rheumatoid arthritis synovialfibroblasts (RA-FLSs). Methods The levels of hsa_circ_0032131 and miR-145-5p in peripheral blood of RA patients and healthy subjects, RA-FLSs as well as normal human synovial fibroblasts were detected. RA-FLSs were divided into control (NC) group, si-NC group, si-hsa_circ_0032131 group, miR-NC group, miR-145-5p group, anti-miR-NC+si-hsa_circ_0032131 group, anti-miR-145-5p+si-hsa_circ_0032131 group, pcDNA group and pcDNA-hsa_circ_0032131 group. CCK-8 and transwell assays were used to detect the viability, migration and invasion of RA-FLSs. The targeting relationship between hsa_circ_0032131 and miR-145-5p was determined through a dual luciferase reporter assay. Results Compared with healthy subjects, the expression of hsa_circ_0032131 was increased, while the expression of miR-145-5p was decreased in the peripheral blood of RA patients, and the difference was statistically significant (P < 0.05). Compared with normal human synovial fibroblasts, the expression of hsa_circ_0032131 was increased, while the expression of miR-145-5p was decreased in RA-FLSs, and the difference was statistically significant (P < 0.05). Compared with the NC group, the viability, migration and the number of RA-FLSs invasion in the si-hsa_circ_0032131 group were reduced, and the difference was statistically significant (P < 0.05). Compared with the miR-NC group, the viability, migration and the number of RA-FLSs invasion in the miR-145-5p group were reduced, and the difference was statistically significant (P < 0.05). Compared with the anti-miR-NC+si-hsa_circ_0032131 group, the viability, migration and the number of RA-FLSs invasion in the anti-miR-145-5p+si-hsa_circ_0032131 group were increased, and the difference was statistically significant (P < 0.05). Compared with the pcDNA group, the level of miR-145-5p in the RA-FLSs of the pcDNA-hsa_circ_0032131 group was reduced (P < 0.05). The miR-145-5p was a direct target of hsa_circ_0032131. Conclusion Knockdown of hsa_circ_0032131 inhibits the proliferation, migration, and invasion of RA-FLSs by upregulating miR-145-5p.

ObjectiveTo explore the effects and possible mechanisms of Jianpi Bushen Formula （健脾补肾方） on radiation-induced immune function damage of mice. MethodsFifty mice were randomly divided into four groups： normal group， model group， thymosin group， high- and low-dose groups of Jianpi Bushen Formula， with 10 mice in each group. Except for the normal group， the mice in the other groups were irradiated with a single whole-body dose of 6.0 Gy X-rays to establish a radiation-injured mouse model. After the successful modeling， the low- and high-dose groups of the Jianpi Bushen Formula were given respectively 13 g/（kg·d）、 26 g/（kg·d） of the formula by gavage， while the thymosin group was given 11.7 mg/（kg·d） of thymosin by gavage， and the normal group and model group were given 0.1 ml/（10g·d） of 0.9% sodium chloride solution by gavage. Each group was administered once a day for 7 consecutive days. After the last gavage， the mice were weighed， and their spleens were separated and weighed to calculate the spleen index. The levels of interferon gamma （IFN-γ） and interleukin 2 （IL-2） in the spleen tissue were detected by enzyme linked immunosorbent assay （ELISA）. The autophagosomes in the spleen were observed by transmission electron microscopy. The mRNA expression levels of phosphatidylinositol 3-kinase （PI3K）， protein kinase B （Akt）， and mammalian target of rapamycin （mTOR） in the spleen were detected by real-time fluorescent quantitative PCR. The protein expression of PI3K， Akt， and mTOR in the spleen， as well as the expression of autophagy-related proteins microtubule-associated light chain protein 3 （LC3）， Beclin1， and p62 were detected by Western blot. ResultsCompared with the normal group， the model group showed significant decreases in body weight， spleen index， and levels of IFN-γ and IL-2 in the spleen （P＜0.01）； the mRNA and protein expression levels of PI3K， Akt， and mTOR in the spleen were also significantly reduced （P＜0.01）； the expression of Beclin1 protein， the ratio of LC3-II/LC3-I significantly increased （P＜0.01）， while the level of p62 protein expression significantly decreased （P＜0.01）. And transmission electron microscopy showed a significant increase in the number of autophagosomes in the spleen and severe cell structure damage in the model group. Compared with the model group， all the above indicators in each medication group were significantly improved （P＜0.05 or P＜0.01）. In the high-dose Jianpi Bushen Formula group， partial intact cristae were visible in the fine mitochondria of the spleen， and there were more autophagosomes. In the low-dose Jianpi Bushen Formula group and thymosin group， the structure of the fine mitochondria in the spleen was relatively intact， and there were fewer autophagosomes. The improvement effect of the low-dose Jianpi Bushen Formula group was better than that of the high-dose group （P＜0.05 or P＜0.01）， and there was no significant difference between the low-dose group and the thymosin group in terms of each indicator （P＞0.05）. ConclusionJianpi Bushen Formula may alleviate the structural damage of the spleen， promote the recovery of immune function， and achieve a best effect at a low dose by enhancing the PI3K/Akt/mTOR signaling pathway in the spleen and inhibiting the over-activation of autophagy induced by radiation.

OBJECTIVE To establish the estimation model for the exposure of mycophenolic acid （MPA） in early renal transplant recipients [calculated by the area under the plasma concentration-time curve with 12 h （AUC0－12 h）]. METHODS Twenty kidney transplant recipients， who received triple immunosuppressive therapy of mycophenolate mofetil （MMF）+tacrolimus+ methylprednisolone， were selected and given MMF dispersible tablets （750 mg， q12 h） on the 15th day after the operation； the blood samples were collected from the patients before and 0.5， 1.0， 1.5， 2.0， 3.0， 4.0， 6.0， 8.0， 12.0 hours after the administration， respectively. The blood concentration of MPA was determined， and the pharmacokinetic parameters of MPA were calculated. The multivariate linear stepwise regression analysis method was used to fit an estimation formula for the finite sampling method suitable for MPA-AUC0－12 h of the recipients. Bland-Altman analysis was used to evaluate the agreement between the estimation formula and the classical pharmacokinetic method. RESULTS The main pharmacokinetic parameters of MPA in 20 renal transplant recipients： c0 was （1.53±0.84） μg/mL， cmax was （12.07±5.97） μg/mL， t1/2 was （5.41±3.67） h， tmax was （1.58±0.75） h， and the average AUC0－12 h calculated by the classical pharmacokinetic method was （33.95±13.40） μg·h/mL. MPA-AUC0－12 h was estimated with sampling points of “4.0， 8.0， 12.0 h”； the simplified calculation formula was AUC0－12 h＝12.058+2.819c4.0+7.045c8.0+ 3.879c12.0 （R 2＝0.934）. The predicted value had a good correlation and consistency with the measured value， and 95.0% of predicted values did not exceed the x±1.96SD （standard deviation） range. CONCLUSIONS The estimation model is established successfully for the exposure of MPA in early renal transplant recipients； the model has better prediction accuracy and fewer sampling points.

ObjectiveTo explore the occupational needs of hand injury patients and the effects of activity-based analysis and occupation-based interventions. MethodsFrom January, 2021 to June, 2022, 212 patients who accepted occupational therapy for wrist-hand injury in Rehabilitation Center of the First Affiliated Hospital of Nanjing Medical University were interviewed using Canadian Occupational Performance Measure (COPM) to collect the activities that they wanted to do, needed to do, were expected to do, but unable to do. Two senior occupational therapists conducted the activity analysis of the summarized needs, identified occupational components, designed a purposeful occupation-based intervention program, and evaluated the performance and satisfaction using COPM. ResultsA cumulative total of 84 occupational needs were proposed, covering five major domains: work, basic activities of daily living, instrumental activities of daily living, leisure and sleep. The activities mostly impaired involved using computer (8.28%), using chopsticks (6.42%), wringing towels (6.25%), travelling on transport (4.90%) and lifting heavy objects (4.73%). The performance and satisfaction improved after treatment (t > 16.572, P < 0.001). ConclusionThe occupational needs are culturally specific for local hand injury patients; the use of an activity-based analysis and occupation-based intervention is effective on meeting the occupational needs.

OBJECTIVE To explore and establish an analysis method for sulfamethoxazole blood concentration in patients with heart transplantation to guide clinical rational drug use. METHODS A new two-dimensional liquid chromatograph (2D-LC-UV) based on column switching technology was used. The on-line solid-phase extraction of sulfamethoxazole in serum was performed using a one-dimensional column Aston SC2(3.5 mm×25 mm, 5 μm), then it was intercepted and retained through the middle column Aston SBR(3.5 mm×10 mm, 5 μm), and the target analytes were completely separated and detected by transfer to the second-dimension column Aston SNX4(4.6 mm×130 mm, 5 μm). The chromatographic conditions were obtained by optimization. The one-dimensional mobile phase was acetonitrile-methanol-H2O(10:10:70), the flow rate was 0.8 mL·min-1; the two-dimensional mobile phase ratio was BPI-1 basic mobile phase-API-3 acidic mobile phase-methanol(20:40:40) the flow rate was 1.2 mL·min-1. The UV detection wavelength was 240 nm. RESULTS The correlation coefficient between the concentration of sulfamethoxazole and peak area was in the range of 9.96‒200.04 μg·mL-1 with R2=0.999 6, it showed a good linear relationship. The intra-day and inter-day precisions(RSD) at low, medium and high concentrations were <15%, and the relative recoveries were between 85%‒115%. The blood concentration of 56 patients in the hospital was measured, and only 30 patients(53.57%) had the blood sample sulfamethoxazole concentration of 100‒150 μg·mL-1, and the rest were not comfortable about the concentration treatment window; there were 0 cases of grade III-IV adverse reactions in the urinary system/blood system/liver, etc. CONCLUSION This method has a simple pre-treatment, high automation, can be sampled in large volumes, and has high accuracy and sensitivity. It can meet the requirements of clinical applications. The research results can provide a methodological reference for clinical therapeutic drug monitoring..

Objective:To screen serum protein markers and evaluate their diagnostic application value in hepatitis B-related acute-on-chronic liver failure (HBV-ACLF).Methods:Serum samples of patients with HBV-ACLF, chronic hepatitis B (CHB) and normal healthy volunteers ( n = 5/group) were determined by cytokine antibody chip in line with the Chinese Diagnostic Standards Study for HBV-ACLF (COSSH-ACLF) cohort. The differentially expressed proteins significance were identified by microarray analysis and prediction. The preliminary serological markers of HBV-ACLF were screened for diagnosis. The potential markers were determined by enzyme-linked immunosorbent assay (ELISA), area under the receiver operating characteristic curve (AUROC) analysis and liver tissue immunohistochemistry for the diagnosis of HBV-ACLF. Student t-test or Mann-Whitney U test were used to compare the continuous measurement data between the two groups, and analysis of variance and Kruskal-Wallis test were used to compare the continuous measurement data between multiple groups. Results:Cytokine antibody chip preliminary screening results showed that the expression levels of these six cytokines, namely, macrophage inflammatory protein 3α (MIP-3α), hepatocyte growth factor, E-selectin, osteopontin, growth differentiation factor 15 and carcinoembryonic antigen-related cellular adhesion molecule 1 were significantly increased in the HBV-ACLF group. Among them, the expression level of MIP-3α was significantly higher in the HBV-ACLF group (99.6 times higher than CHB group and 146.9 times higher than healthy volunteers’ group, respectively, P < 0.0001) as validated by serum ELISA in 132 HBV-ACLF cases, 91 CHB cases and 72 healthy volunteers. AUROC analysis showed that the high expression of MIP-3α could be used as a marker to distinguish patients with HBV-ACLF from CHB. The AUROC was 0.995 (95% CI: 0.990 ~ 1.000), with sensitivity and specificity of 95.5% and. 98.9%, respectively. Immunohistochemistry showed that MIP-3α was positively expressed in HBV-ACLF-derived liver tissues, and negatively expressed in CHB-derived liver and normal liver tissues. Conclusion:Serum MIP-3α level is closely related to the pathological characteristics of HBV-ACLF. Therefore, it may be used as a potential serological marker for the diagnosis of HBV-ACLF.