Due to the diverse composition and complex physicochemical and biological characteristics, the pre-treatment of microbiological counting method （preparation of test solution） in microbiological limit test were interfered by many factors, which ultimately affected the repeatability and accuracy of test results. Improving the accuracy of microbiological test is of practical significance to ensure the safety and effectiveness of non-sterile preparations. In this paper, the key factors and optimization methods involved in the pre-treatment of proprietary Chinese medicines were systematically analyzed and summarized.

Subtype classification of age-related macular degeneration (AMD) based on optical coherence tomography (OCT) images serves as an effective auxiliary tool for clinicians in diagnosing disease progression and formulating treatment plans. To improve the classification accuracy of AMD subtypes, this study proposes a keypoint-based, attention-enhanced residual network (KPA-ResNet). The proposed architecture adopts a 50-layer residual network (ResNet-50) as the backbone, preceded by a keypoint localization module based on heatmap regression to outline critical lesion regions. A two-dimensional relative self-attention mechanism is incorporated into convolutional layers to enhance the representation of key lesion areas. Furthermore, the network depth is appropriately increased and an improved residual module, ConvNeXt, is introduced to enable comprehensive extraction of high-dimensional features and enrich the detail of lesion boundary contours, ultimately achieving higher classification accuracy of AMD subtypes. Experimental results demonstrate that KPA-ResNet achieves significant improvements in overall classification accuracy compared with conventional convolutional neural networks. Specifically, for the wet AMD subtypes, the classification accuracies for inactive choroidal neovascularization (CNV) and active CNV reach 92.8% and 95.2%, respectively, representing substantial improvement over ResNet-50. These findings validate the superior performance of KPA-ResNet in AMD subtype classification tasks. This work provides a high-accuracy, generalizable network architecture for OCT-based AMD subtype classification and offers new insights into integrating attention mechanisms with convolutional neural networks in ophthalmic image analysis.
Tomography, Optical Coherence/methods* ; Humans ; Macular Degeneration/diagnostic imaging* ; Neural Networks, Computer

The spatio-temporal heterogeneity of breast cell subsets forms the fundamental biological basis for physiological development and pathological progression, including tumorigenesis; however, its complex regulatory mechanisms are not yet fully elucidated. With its high-resolution capabilities, single-cell RNA sequencing (scRNA-seq) technology offers a powerful tool for dissecting this cellular heterogeneity. This technology enables the construction of high-precision breast cell atlases, the accurate identification of distinct cell subsets, and the reconstruction of differentiation trajectories from stem/progenitor cells to functional epithelial cells. By resolving the transcriptional regulatory networks that govern cell fate determination, intercellular communication patterns, and dynamic microenvironmental interactions, scRNA-seq has unveiled the molecular foundations of breast development and provided new perspectives on the pathogenesis of related diseases such as breast cancer and macromastia. Furthermore, scRNA-seq demonstrates significant potential for discovering early molecular markers of disease, deciphering tumor heterogeneity, and elucidating mechanisms of therapeutic resistance. The continued application of scRNA-seq for dissecting breast cell heterogeneity, combined with its integration with multi-modal data such as spatial omics, promises to provide critical evidence and new insights for revealing the molecular mechanisms of breast development-related diseases and for formulating precision therapeutic strategies.
Humans ; Single-Cell Analysis/methods* ; Female ; Breast Neoplasms/pathology* ; Sequence Analysis, RNA/methods* ; Breast/cytology*

Somatostatin receptor 1 (SSTR1) is a crucial therapeutic target for various neuroendocrine and oncological disorders. Current SSTR1-targeted treatments, including the first-generation somatostatin analog lanreotide (Lan) and the second-generation analog pasireotide (Pas), show promise but encounter challenges related to selectivity and efficacy. This study presents high-resolution cryo-electron microscopy structures of SSTR1 complexed with Lan or Pas, revealing the distinct mechanisms of ligand-binding and activation. These structures illustrate unique conformational changes in the SSTR1 orthosteric pocket induced by each ligand, which are critical for receptor activation and ligand selectivity. Combined with the biochemical assays and molecular dynamics simulations, our results provide a comparative analysis of binding characteristics within the SSTR family, highlighting subtle differences in SSTR1 activation by Lan and Pas. These insights pave the way for designing next-generation therapies with enhanced efficacy and reduced side effects through improved receptor subtype selectivity.

Objective To investigate the effects of Yitangkang on renal ectopic lipid deposition and the PPAR-α/PGC-1α signaling pathway in db/db mice;To explore its mechanism in improving renal injury.Methods Totally 30 db/db mice were divided into model group,Yitangkang group and losartan potassium group(10 mice in each group)using a random number table method.An additional 10 db/m mice were assigned as the blank group.The mice received corresponding interventions for 8 weeks.Body mass,fasting blood glucose(FBG),serum creatinine(SCr),blood urea nitrogen(BUN),and 24-hour urinary protein content were measured,renal morphology and injury were observed using HE,PAS and Masson staining,lipid deposition in renal tissue was observed by oil red O staining,renal ultrastructure was observed by transmission electron microscopy,immunofluorescence staining was used to detect the expressions of CPT1A and Nrf2 in renal tissue,RT-qPCR was performed to assess the mRNA expressions of PPAR-α and PGC-1α,Western blot was used to detect the protein expressions of PPAR-α,CPT1A,PGC-1α,Nrf2,NRF1 and TFAM.Results Compared with the blank group,the body mass and FBG significantly increased in the model group(P<0.05),and the contents of SCr,BUN and 24-hour urinary protein significantly increased(P<0.05);histopathology revealed glomerular hypertrophy,mesangial cell and matrix proliferation,thickened basement membrane,abnormal deposition of interstitial collagen fibers,increased lipid deposition in renal tissue,widespread foot process effacement,reduced foot process density(P<0.05),blurred mitochondrial outer membranes,swollen morphology,and indistinct cristae;mRNA expressions of PPAR-α and PGC-1α significantly decreased(P<0.05),and protein expressions of PPAR-α,CPT1A,PGC-1α,Nrf2,NRF1 and TFAM significantly decreased(P<0.05).Compared with the model group,the body mass and FBG of mice in Yitangkang group significantly decreased(P<0.05),but there was no significant difference in the above indexes in losartan potassium group(P>0.05);the contents of SCr,BUN and 24-hour urinary protein in Yitangkang group and losartan potassium group significantly decreased(P<0.05);the pathological damage and lipid deposition of renal tissue was alleviated,the ultrastructure of podocytes was improved,and the density of podocytes significantly increased(P<0.05);the mRNA expressions of PPAR-α and PGC-1α in renal tissue increased,and the protein expressions of PPAR-α,CPT1A,PGC-1α,Nrf2,NRF1 and TFAM increased,with statistical significance(P<0.05).Conclusion Yitangkang can effectively alleviate renal injury in diabetic nephropathy mice.The mechanism may be related to the activation of PPAR-α/PGC-1α signaling pathway and the reduction of renal lipid deposition and mitochondrial biogenesis.

Reduced or interrupted blood flow is an important pathological and physiological process in femoral head necrosis,so understanding the blood flow of the femoral head can help better understand the progression of femoral head necrosis.With the continuous development and improvement of imaging technology,the technical methods for detecting the blood flow of the femoral head are gradually being widely applied,allowing clinical physicians to better understand the blood flow situation of patients with femoral head necrosis.However,at present,the prognosis prediction of femoral head necrosis still mainly revolves around factors such as the area and angle of femoral head necrosis.Therefore,this article explores imaging techniques covering studies such as femoral head vascular imaging or blood flow perfusion parameters,summarizes their application research progress in femoral head necrosis blood flow assessment,and aims to provide more objective basis for exploring the prevention and prognosis of femoral head necrosis from the perspective of femoral head blood flow.

Objective:To explore the application of Rain Classroom combined with BOPPPS teaching mode in the teaching of Obstetrics and Gynecology Nursing.Methods:A total of 184 undergraduate nursing students in 6 classes of grade 2022 at a medical college were enrolled.Using cluster sampling,92 students from 3 classes were randomly assigned to the control group and another 92 students were selected as the experimental group.The control group received traditional teaching methods,while the experimental group received the Rain Classroom combined with BOPPPS teaching mode.Autonomous learning ability,critical thinking ability,teaching effectiveness evaluations,formative assessments,and final examination scores were compared between the two groups.Result:The scores of autonomous learning ability,critical thinking ability,teaching effectiveness evaluations,formative assessments,and final examination in the experimental group were higher than those in the control group after teaching(P＜0.05).Conclusion:Rain Classroom combined with BOPPPS teaching mode has outstanding teaching effect in the teaching of Obstetrics and Gynecology Nursing,cultivating students'autonomous learning ability and critical thinking ability,and improving students'participation in the whole process.

Objective·To determine hyaluronan(HA)expression in the endocrine-resistant microenvironment of estrogen receptor-positive(ER+)breast cancer and elucidate its impact on the acquired resistance.Methods·Chemiluminescent immunoassay was used to quantify HA levels in the culture supernatants of fulvestrant-resistant breast cancer cells.An immunofluorescence(IF)assay was performed to visualize the colocalization of CD44 and HA in MCF7/FulR cells.Using an established adaptive endocrine-resistant breast cancer mouse model,HA expression in resistant breast cancer tissues was assessed by immunohistochemistry(IHC)assay.Single-cell RNA sequencing(scRNA-seq)and RNA sequencing(RNA-seq)were conducted to examine transcriptomic profiles and alterations in HA-related genes in resistant breast cancer cells.Flow cytometry(FCM)was utilized to measure the proportion of CD44+CD24-cells in MCF7/FulR.The correlation between HA synthesis genes and cell stemness was investigated in clinical ER+breast cancers from GEO data sets.Hyaluronidase(HAase)treatment was applied to remove the"HA coat",and RT-qPCR and Western blotting analysis were carried out to monitor changes in stemness-related molecules.CCK-8 assays,flow cytometry(FCM),and Hoechst 33258 staining were performed to determine changes in apoptosis and fulvestrant efficiency after HAase treatment.Results·IF results revealed that compared with MCF7 cells,the"HA coat"on the surface of MCF7/FulR cells was significantly thickened.IHC demonstrated markedly increased HA retention in fulvestrant-resistant mouse breast cancer tissues.ScRNA-seq and RNA-seq analyses indicated elevated expression of stemness-related genes and HA synthesis-associated genes in fulvestrant-resistant breast cancer cells.Correlation analysis revealed a positive association between HA synthesis and cancer stemness in ER+breast cancer.IF and RT-qPCR results demonstrated that removing the HA coating from the surface of MCF7/FulR cells led to a significant reduction in the expression of stemness-related molecules;concurrently,CCK-8 assays,FCM analysis,and Hoechst 33258 staining revealed that"HA coat"clearance reduced MCF7/FulR'tolerance to fulvestrant and increased apoptosis.Conclusion·Endocrine-resistant breast cancer cells develop an enriched"HA coat",which promotes stemness in fulvestrant-resistant tumors.Disruption of this HA coat through HAase treatment effectively reduces cell stemness,induces apoptosis,and re-sensitizes breast cancer cells to fulvestrant.

Objective·To determine hyaluronan(HA)expression in the endocrine-resistant microenvironment of estrogen receptor-positive(ER+)breast cancer and elucidate its impact on the acquired resistance.Methods·Chemiluminescent immunoassay was used to quantify HA levels in the culture supernatants of fulvestrant-resistant breast cancer cells.An immunofluorescence(IF)assay was performed to visualize the colocalization of CD44 and HA in MCF7/FulR cells.Using an established adaptive endocrine-resistant breast cancer mouse model,HA expression in resistant breast cancer tissues was assessed by immunohistochemistry(IHC)assay.Single-cell RNA sequencing(scRNA-seq)and RNA sequencing(RNA-seq)were conducted to examine transcriptomic profiles and alterations in HA-related genes in resistant breast cancer cells.Flow cytometry(FCM)was utilized to measure the proportion of CD44+CD24-cells in MCF7/FulR.The correlation between HA synthesis genes and cell stemness was investigated in clinical ER+breast cancers from GEO data sets.Hyaluronidase(HAase)treatment was applied to remove the"HA coat",and RT-qPCR and Western blotting analysis were carried out to monitor changes in stemness-related molecules.CCK-8 assays,flow cytometry(FCM),and Hoechst 33258 staining were performed to determine changes in apoptosis and fulvestrant efficiency after HAase treatment.Results·IF results revealed that compared with MCF7 cells,the"HA coat"on the surface of MCF7/FulR cells was significantly thickened.IHC demonstrated markedly increased HA retention in fulvestrant-resistant mouse breast cancer tissues.ScRNA-seq and RNA-seq analyses indicated elevated expression of stemness-related genes and HA synthesis-associated genes in fulvestrant-resistant breast cancer cells.Correlation analysis revealed a positive association between HA synthesis and cancer stemness in ER+breast cancer.IF and RT-qPCR results demonstrated that removing the HA coating from the surface of MCF7/FulR cells led to a significant reduction in the expression of stemness-related molecules;concurrently,CCK-8 assays,FCM analysis,and Hoechst 33258 staining revealed that"HA coat"clearance reduced MCF7/FulR'tolerance to fulvestrant and increased apoptosis.Conclusion·Endocrine-resistant breast cancer cells develop an enriched"HA coat",which promotes stemness in fulvestrant-resistant tumors.Disruption of this HA coat through HAase treatment effectively reduces cell stemness,induces apoptosis,and re-sensitizes breast cancer cells to fulvestrant.

Objective:To explore the application of Rain Classroom combined with BOPPPS teaching mode in the teaching of Obstetrics and Gynecology Nursing.Methods:A total of 184 undergraduate nursing students in 6 classes of grade 2022 at a medical college were enrolled.Using cluster sampling,92 students from 3 classes were randomly assigned to the control group and another 92 students were selected as the experimental group.The control group received traditional teaching methods,while the experimental group received the Rain Classroom combined with BOPPPS teaching mode.Autonomous learning ability,critical thinking ability,teaching effectiveness evaluations,formative assessments,and final examination scores were compared between the two groups.Result:The scores of autonomous learning ability,critical thinking ability,teaching effectiveness evaluations,formative assessments,and final examination in the experimental group were higher than those in the control group after teaching(P＜0.05).Conclusion:Rain Classroom combined with BOPPPS teaching mode has outstanding teaching effect in the teaching of Obstetrics and Gynecology Nursing,cultivating students'autonomous learning ability and critical thinking ability,and improving students'participation in the whole process.