ObjectiveTo establish an in vitro culture model of rat corpora cavernous smooth muscle cells （CCSMCs） using a modified tissue block adherence method. MethodsCorpus cavernosum smooth muscle tissue was digested with collagenase type I and subsequently cultured using an adherent method. Cells were purified via differential adhesion and identified through immunofluorescence and Western blotting. ResultsCCSMCs began to emerge from the tissue block after 3 days， increased significantly by day 7， and converged by day 12. Post-passage， CCSMCs exhibited strong proliferation and a “peak-to-valley” phenomenon. After purification， the cells tested positive for α-smooth muscle actin （α-SMA）， confirming the successful establishment of the in vitro culture model. ConclusionThe modified tissue block adherence method is a cost-effective and efficient way to obtain high-purity CCSMCs.

Andrological diseases have become an important public health problem threatening men's health worldwide， which significantly affects the quality of life of patients and brings a heavy disease burden. Western medicine often faces the dilemma of obvious side effects and limited efficacy. Traditional Chinese medicine has unique advantages in the prevention and treatment of andrological diseases and has accumulated rich clinical experience. Epimedii Folium， as a traditional Chinese medicine for strengthening kidney and Yang， exerts a key therapeutic effect on andrology diseases through multi-component synergy， multi-target regulation， and multi-pathway intervention. Recent studies have found that the main active components of Epimedii Folium， such as icariin， icariside， and icaritin， are the key material basis for the treatment of andrological diseases. The active components of Epimedii Folium can play a role in common andrological diseases such as erectile dysfunction， male infertility， and prostate cancer by regulating the activity of the nitric oxide/cyclic guanosine monophosphate （NO/cGMP） pathway， participating in oxidative stress response， regulating the secretion of hypothalamic-pituitary-gonadal axis hormones， improving spermatogenic dysfunction， and inhibiting the proliferation of cancer cells. However， the systematic action network and molecular mechanisms of the active components of Epimedii Folium have not been fully elucidated， thereby limiting its potential for clinical translation and application. In the future， it is necessary to combine cutting-edge technologies such as metabolomics， single-cell sequencing， and targeted nanoscale drug delivery systems， strengthening the research on the compatibility rules of active components and organ-specific delivery， providing a scientific basis for the development of innovative andrology traditional Chinese medicine formulas with international competitiveness， and promoting the innovation and breakthrough of andrology disease treatment modes.

A comprehensive phytochemical investigation of the leaves and twigs of Physalis angulata. var. villosa resulted in the isolation of 23 withanolide derivatives, including one novel 13,20-γ-lactone withanolide derivative (1) and three new withanolide derivatives (2-4). Architecturally, physalinin A (1) represents the first identified type B withanolide featuring a 13,20-γ-lactone moiety. The molecular structures of all isolates were elucidated using an integrated approach combining nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry (MS), infrared (IR) spectroscopy, and quantum chemical calculations to confirm structural assignments. The antiproliferative activities of all isolated withanolides were evaluated against four human cancer cell lines (HEL, HCT-116, Colo320DM, and MDA-MB-231). Among them, eight derivatives (2, 5-8, 14, 15, and 23) exhibited significant inhibitory effects, with half-maximal inhibitory concentration (IC₅₀) values of 0.18 ± 0.03 to 17.02 ± 0.21 μmol·L^-1. Structure-activity relationship (SAR) analysis suggested that the presence of an epoxide ring enhances anticancer activity, potentially through increased reactivity or specific interactions with molecular targets involved in cancer progression. These findings underscore the pharmacological potential of withanolides as promising lead compounds for the development of novel anticancer therapeutics.
Withanolides/isolation & purification* ; Physalis/chemistry* ; Humans ; Molecular Structure ; Cell Line, Tumor ; Antineoplastic Agents, Phytogenic/isolation & purification* ; Cell Proliferation/drug effects* ; Plant Leaves/chemistry* ; Plant Extracts/pharmacology*

Objective To investigate the early diagnostic value of combined detection of serum TIR-domain-containing adapter-inducing interferon-β(TRIF),cyclophilin A(CyPA),ubiquitin carboxyl-terminal hydrolase L1(UCH-L1),and hepcidin(Hepc)in brain injury among premature infants(BIPI).Methods Clinical data from 105 infants with brain injury following perinatal asphyxia(BIPI)admitted to our hospital between January 2022 and December 2024 were retrospectively collected and assigned to the case group,which was further subdivided into a severe subgroup(n=31)and a mild subgroup(n=74)based on the extent of brain injury.As controls,clinical data from 105 healthy preterm infants born in our hospital during the same period were retrospectively enrolled in a 1∶1 ratio.Serum levels of TRIF,CyPA,UCH-L1,and Hepc,along with clinical characteristics,were compared between the groups.Pearson correlation analysis was performed to evaluate the association between serum biomarker levels and neonatal behavioral neurological assessment(NBNA)scores.The diagnostic performance of these bio-markers for early detection of BIPI was assessed by constructing receiver operating characteristic(ROC)curves and calculating the area under the curve(AUC).Results Serum levels of TRIF,CyPA,UCH-L1,and Hepc were significantly higher in the case group than in the control group(P<0.05),while NBNA scores were significantly lower in the case group(P<0.05).Similarly,serum levels of these biomarkers were significantly elevated in the severe group compared to the mild group(P<0.05),accompanied by significantly lower NBNA scores in the severe group(P<0.05).Pearson correlation analysis revealed that serum levels of TRIF,CyPA,UCH-L1,and Hepc were negatively correlated with NBNA scores(r=-0.579,-0.514,-0.609,-0.588;all P<0.05).ROC analysis demon-strated that the combined detection of these four markers yielded an AUC of 0.927 for early diagnosis of BIPI,which was significantly higher than the AUCs of individual markers(0.819,0.803,0.776,and 0.767,respectively;P<0.05).Conclusions The serum levels of TRIF,CyPA,UCH-L1,and Hepc in children with BIPI were significantly elevated.These biomarkers were closely associated with disease progression and neurological development,and their combined measurement demonstrated superior performance in the early diagnosis of BIPI.

Objective To explore the effects of graphene quantum dots(GQDs)on the reprogramming of Müller cells.Methods Müller cells were randomly divided into the control,dedifferentiation,dedifferentiation+GQD,and GQD groups.The cells in the dedifferentiation group and the dedifferentiation+GQD group were dedifferentiated using the serum-free medium containing DMEM/F12(1∶1),20 μg·L-1 EGF,10 μg·L-1 bFGF,2 × B27,and 1 × N2 for 5 d.Then,the cells in the dedifferentiation+GQD group and the GQD group were treated with 50 mg·L-1 GQDs for 72 h.The Müller cells in the control group were subjected to normal cell culture.Immunofluorescence staining was used to identify the ex-pression of Müller cell markers,including glial fibrillary acidic protein(GFAP),glutamate-aspartate transporter(GLAST),and glutamine synthetase(GS).Phalloidin staining was performed to observe whether Müller cells could take up GQDs.The effect of different concentrations of GQDs on the proliferation of Müller cells was assessed using the cell counting kit-8(CCK-8)assay.The effect of GQDs on the expression of neural progenitor/stem cell markers(SRY-box transcription factor 2[SOX-2]and Nestin)and the astrocyte marker GFAP in normal and dedifferentiated Müller cells was examined using im-munofluorescence staining and Western blotting.Results The immunofluorescence staining results showed that the fluo-rescence of GFAP was extremely weak and almost invisible in Müller cells,while the fluorescence of GLAST and GS was extremely strong and predominantly appeared in the cytoplasm of Müller cells.After 24 h of GQD treatment,trace amounts of GQD fluorescence were visible in Müller cells.The amount of GQD fluorescence in the cytoplasm of Müller cells gradual-ly increased with time.The CCK-8 assay results showed that the activity of Müller cells tended to decrease with an increase in the treatment time and concentration of GQD.The statistical analysis results showed that the mean fluorescence intensity of GFAP,Nestin,and SOX-2 in Müller cells of the dedifferentiation,dedifferentiation+GQD,and GQD groups was higher than that of the control group,and the differences were statistically significant(all P＜0.05).The mean fluorescence inten-sity of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the mean fluores-cence intensity of Nestin and SOX-2 was lower than that in the dedifferentiation group,and the differences were all statisti-cally significant(P＜0.05).The relative protein expression of GFAP,Nestin,and SOX-2 in Müller cells in the dedifferentia-tion,dedifferentiation+GQD,and GQD groups was higher than that in the control group,and the differences were statis-tically significant(all P＜0.05).The relative protein expression of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the relative protein expression of Nestin and SOX-2 was lower than that in the dedi-fferentiation group,and the differences were statistically significant(all P＜0.05).Conclusion GQDs facilitate the re-programming of Müller cells into astrocytes.

Objective To investigate the early diagnostic value of combined detection of serum TIR-domain-containing adapter-inducing interferon-β(TRIF),cyclophilin A(CyPA),ubiquitin carboxyl-terminal hydrolase L1(UCH-L1),and hepcidin(Hepc)in brain injury among premature infants(BIPI).Methods Clinical data from 105 infants with brain injury following perinatal asphyxia(BIPI)admitted to our hospital between January 2022 and December 2024 were retrospectively collected and assigned to the case group,which was further subdivided into a severe subgroup(n=31)and a mild subgroup(n=74)based on the extent of brain injury.As controls,clinical data from 105 healthy preterm infants born in our hospital during the same period were retrospectively enrolled in a 1∶1 ratio.Serum levels of TRIF,CyPA,UCH-L1,and Hepc,along with clinical characteristics,were compared between the groups.Pearson correlation analysis was performed to evaluate the association between serum biomarker levels and neonatal behavioral neurological assessment(NBNA)scores.The diagnostic performance of these bio-markers for early detection of BIPI was assessed by constructing receiver operating characteristic(ROC)curves and calculating the area under the curve(AUC).Results Serum levels of TRIF,CyPA,UCH-L1,and Hepc were significantly higher in the case group than in the control group(P<0.05),while NBNA scores were significantly lower in the case group(P<0.05).Similarly,serum levels of these biomarkers were significantly elevated in the severe group compared to the mild group(P<0.05),accompanied by significantly lower NBNA scores in the severe group(P<0.05).Pearson correlation analysis revealed that serum levels of TRIF,CyPA,UCH-L1,and Hepc were negatively correlated with NBNA scores(r=-0.579,-0.514,-0.609,-0.588;all P<0.05).ROC analysis demon-strated that the combined detection of these four markers yielded an AUC of 0.927 for early diagnosis of BIPI,which was significantly higher than the AUCs of individual markers(0.819,0.803,0.776,and 0.767,respectively;P<0.05).Conclusions The serum levels of TRIF,CyPA,UCH-L1,and Hepc in children with BIPI were significantly elevated.These biomarkers were closely associated with disease progression and neurological development,and their combined measurement demonstrated superior performance in the early diagnosis of BIPI.

Objective To explore the effects of graphene quantum dots(GQDs)on the reprogramming of Müller cells.Methods Müller cells were randomly divided into the control,dedifferentiation,dedifferentiation+GQD,and GQD groups.The cells in the dedifferentiation group and the dedifferentiation+GQD group were dedifferentiated using the serum-free medium containing DMEM/F12(1∶1),20 μg·L-1 EGF,10 μg·L-1 bFGF,2 × B27,and 1 × N2 for 5 d.Then,the cells in the dedifferentiation+GQD group and the GQD group were treated with 50 mg·L-1 GQDs for 72 h.The Müller cells in the control group were subjected to normal cell culture.Immunofluorescence staining was used to identify the ex-pression of Müller cell markers,including glial fibrillary acidic protein(GFAP),glutamate-aspartate transporter(GLAST),and glutamine synthetase(GS).Phalloidin staining was performed to observe whether Müller cells could take up GQDs.The effect of different concentrations of GQDs on the proliferation of Müller cells was assessed using the cell counting kit-8(CCK-8)assay.The effect of GQDs on the expression of neural progenitor/stem cell markers(SRY-box transcription factor 2[SOX-2]and Nestin)and the astrocyte marker GFAP in normal and dedifferentiated Müller cells was examined using im-munofluorescence staining and Western blotting.Results The immunofluorescence staining results showed that the fluo-rescence of GFAP was extremely weak and almost invisible in Müller cells,while the fluorescence of GLAST and GS was extremely strong and predominantly appeared in the cytoplasm of Müller cells.After 24 h of GQD treatment,trace amounts of GQD fluorescence were visible in Müller cells.The amount of GQD fluorescence in the cytoplasm of Müller cells gradual-ly increased with time.The CCK-8 assay results showed that the activity of Müller cells tended to decrease with an increase in the treatment time and concentration of GQD.The statistical analysis results showed that the mean fluorescence intensity of GFAP,Nestin,and SOX-2 in Müller cells of the dedifferentiation,dedifferentiation+GQD,and GQD groups was higher than that of the control group,and the differences were statistically significant(all P＜0.05).The mean fluorescence inten-sity of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the mean fluores-cence intensity of Nestin and SOX-2 was lower than that in the dedifferentiation group,and the differences were all statisti-cally significant(P＜0.05).The relative protein expression of GFAP,Nestin,and SOX-2 in Müller cells in the dedifferentia-tion,dedifferentiation+GQD,and GQD groups was higher than that in the control group,and the differences were statis-tically significant(all P＜0.05).The relative protein expression of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the relative protein expression of Nestin and SOX-2 was lower than that in the dedi-fferentiation group,and the differences were statistically significant(all P＜0.05).Conclusion GQDs facilitate the re-programming of Müller cells into astrocytes.

Objective:To explore the clinical efficacy and safety analysis of a novel laser localization technology assisted percutaneous puncture of trigeminal nerve microsphere capsule compression surgery for the treatment of primary trigeminal neuralgia.Methods:A retrospective selection was conducted on 63 patients with primary trigeminal neuralgia who underwent percutaneous puncture of the trigeminal nerve microsphere capsule compression surgery at the First Hospital of Hunan University of Chinese Medicine from January 2020 to December 2021. According to different surgical methods, they were divided into a new laser localization assisted puncture group (observation group) of 32 cases and a traditional barehanded localization puncture group (control group) of 31 cases. An analysis was conducted on the surgical time, puncture time, puncture frequency, intraoperative exposure to radiation, number of cases of poor balloon formation, and clinical efficacy within 6 months after surgery for two groups of patients. The prognosis of the patients was followed up at 6 months after surgery.Results:The surgical time, puncture time, puncture frequency, and intraoperative exposure of the observation group were all less than those of the control group, and the differences were statistically significant (all P<0.05). There was no statistically significant difference ( P>0.05) in the number of cases of poor balloon angioplasty between the observation group and the control group, as well as the pain score grading of the Barlow Neurological Institute (BNI) on the first day after surgery. Within 6 months after surgery, there was no statistically significant difference in the incidence of facial numbness, diplopia, masseter weakness, perilabial herpes, and recurrent pain between the two groups of patients (all P>0.05). Conclusions:Laser positioning technology can assist in precise puncture of the foramen ovale and accurate placement of balloons based on surgical experience, which helps to improve surgical safety, reduce postoperative complications and intraoperative radiation dose, and achieve satisfactory short-term follow-up results.

The key pathogenesis of coronary heart disease （CHD） is spleen deficiency and phlegm stasis， and dysfunctional high-density lipoprotein （dys-HDL） may be the biological basis for the occurrence of CHD due to spleen deficiency and phlegm stasis. Considering the biological properties and effects of high-density lipoprotein （HDL）， it is believed that the structure and components of HDL are abnormal in the state of spleen deficiency which led to dys-HDL； and dys-HDL contributes to the formation of atherosclerotic plaques through two major pathways， namely， mediating the dysfunction of endothelial cells and mediating the foaminess of macrophages and smooth muscle cells， thus triggering the development of CHD. It is also believed that dys-HDL is a microcosmic manifestation and a pathological product of spleen deficiency， and spleen deficiency makes foundation for the production of dys-HDL； dys-HDL is also an important biological basis for the phlegm-stasis interactions in CHD. The method of fortifying spleen， resolving phlegm， and dispelling stasis， is proposed as an important principle in the treatment of CHD by traditional Chinese medicine， which can achieve the therapeutic purpose by affecting the changes in the structure and components of dys-HDL， thus revealing the scientific connotation of this method， and providing ideas for the diagnosis and treatment of CHD by traditional Chinese medicine.

Objective To explore the clinical efficacy and safety of donepezil(DNPQ)combined with butylphthalide sequential therapy(BST)in the treatment of Parkinson's syndrome(PS).Methods In this study,104 patients with Parkinson's disease(PD)who were diagnosed and treated in the Department of Neurology of The Fourth Hospital of Changsha from January 2020 to November 2023 were randomly divided into a control group(butylphthalide softcapsule combined with DNPQ)and an observation group(BST combined with DNPQ).The main observation indicators of this study were the clinical efficacy and drug-related adverse reactions after 3 months of treatment.The secondary observation indicators were the cognitive function[Montreal Cognitive Assessment(MoCA)and Mini-Mental State Examination(MMSE)],overall condition[Unified Parkinson's Disease Rating Scale(UPDRS)],activity of daily living(ADL),and oxidative stress-related cytokines[recombinant human Parkinson's disease protein 7(PARK7),neurotrophic factor 3(NT3),and C-reactive protein(CRP)]improvement after treatment.Results There were 52 patients in the experimental group and 52 patients in the control group.The treatment efficacy rate in the experimental group was significantly higher than that in the control group(P<0.05),while the incidence of adverse reactions was significantly lower than that in the control group(P<0.05).Before treatment,there were no significant differences in MoCA scores,MMSE scores,UPDRS scores,ADL scores,serum NT3,CRP,and PARK7 levels between the two groups(P>0.05).After treatment,the MoCA score,MMSE score,and ADL score in the experimental group were higher than those in the control group(P<0.05),while the UPDRS score was lower than that in the control group(P<0.05).After treatment,the serum NT3 level in the experimental group was higher than that in the control group(P<0.05),while the serum CRP and PARK7 levels were lower than those in the control group(P<0.05).Conclusion The combination of DNPQ and BST has better clinical efficacy and safety,which can improve cognitive function,ADL and oxidative stress-related cytokine content in patients with PS.