BACKGROUND:In vitro reconstruction of hair follicles depends on the self-assembly behavior of dermal papilla cells and keratinocytes during three dimensional co-culture,which is very similar to the development of hair follicle embryos.Therefore,hair follicle organoids can be used as a good model in vitro for basic research of hair follicle regeneration.Currently,wholly humanized follicle organoids cultured by biomimetic system have been successfully transplanted into animals.Research on hair follicle organoids makes it possible to regenerate hair follicles,but there is no uniform conclusion on the construction method,and there are still many challenges.OBJECTIVE:To review the current research progress of hair follicle organoids,summarize their culture methods and characterization,and expound their application prospects,so as to provide reliable basis for subsequent research,in order to promote its clinical transformation in tissue engineering and alopecia treatment.METHODS:CNKI and PubMed databases were searched by a computer,using"hair follicle regeneration,hair regeneration,organoid"as Chinese search terms and"hair growth,hair loss,hair regeneration,hair follicle,dermal papilla cell,organoid"as English search terms.The domestic and foreign literature on hair follicle organoids was reviewed and a total of 67 articles were included for follow-up analysis.RESULTS AND CONCLUSION:(1)Dermal papilla cells and keratinocytes from different sources can be used to construct hair follicle organoids in vitro to obtain mouse-derived,human-mouse chimeric and human-derived hair follicle organoids,which have been successfully used in rodent transplantation models.(2)A variety of programming methods,such as electrical stimulation,addition of recombinant protein,and dissociation of cell gene expression,can be used to promote hair follicle organoid such ashair follicle germ formation and hair shaft elongation.Simultaneously,the application of various 3D printing technologies has achieved controllable hair follicle organoid to the size of the quantity production,which greatly improves the production efficiency.(3)When melanocytes are added to the dissociated cell co-culture system,the problem of pigment deficiency in new hair follicles may be solved.When human umbilical vein endothelial cells are added,the new blood vessels in the cell aggregates can improve their nutrient supply and reduce necrosis in the central area.(4)As the technology of engineered hair follicle organoid gradually maturates,hair follicle organoids constructed from patient's autologous cells may be used for individualized treatment of severe alopecia.

This study was aimed at comparing the effects of storage conditions on the Brucella survival number in brain heart infu-sion medium,to provide experimental data support for the selection of storage conditions for Brucella strains.We created bacterial sus-pensions of seven strains of Brucella melitensis,Brucella abortus,and Brucella suis(denoted A,B,C,D,E,F,G)by using 30%glycerol brain heart infusion medium,then conducted stability testing.For accelerated stability tests,the bacterial solution was placed in a 37℃chamber for 1,2,3,or 4 weeks continuously,and the Brucella survival number at each time point was measured.For freeze-thaw stability testing,the bacterial solutions were frozen at-80℃,transferred to room temperature for 4 hours,and melted in one freeze-thaw cycle,and the Brucella survival number was measured for 2,4,6,and 8 freeze-thaw times.For long-term stability testing,the bacterial solutions were stored at-80℃,and the Brucella survival number was measured at various time points during the first,third,sixth,and twelfth months of storage.A line graph was plotted to demonstrate the changes in Brucella survival number at each time point in the three stability tests.IBM SPSS Statistics 22 software was used to assess statistically significant differences in Brucella survival number at each time point and the initial Brucella survival number.Accelerated stability tests showed that the Bru-cella survival number of the seven strains significantly decreased,and a statistically significant difference in Brucella survival number was observed after 1 week.In freeze-thaw stability tests,with increasing freeze-thaw times,the Brucella survival number of the seven strains showed a decreasing trend.Strains C and F showed statistically significant differences with respect to the initial Brucella sur-vival number after two times of freeze-thaw;strains A and D showed statistically significant differences with respect to the initial Bru-cella survival number after four times of freeze-thaw;strain G showed statistically significant differences with respect to the initial Bru-cella survival number after six times of freeze-thaw;and strain E showed statistically significant differences with respect to the initial Brucella survival number after eight times of freeze-thaw.The data for strain B showed abnormalities.In long-term stability testing,the Brucella survival number for strains C and F after storage for 1 month showed a statistically significant difference with respect to the initial bacterial solution.No statistically significant difference in Brucella survival number was observed for strains B and E after stor-age for 1,3,6,or 12 months.The data for strains A,D,and G showed abnormalities.The sterile 30%glycerol brain heart infusion medium maintained the Brucella survival number in an-80℃environment,and freezing and thawing multiple times should be avoided.Compared with the traditional freeze-drying method,this method is not only easy to perform but also avoids potential bio-safety hazards,and provides a reliable Brucella culture preservation scheme for researchers in related fields.

This study was aimed at comparing the effects of storage conditions on the Brucella survival number in brain heart infu-sion medium,to provide experimental data support for the selection of storage conditions for Brucella strains.We created bacterial sus-pensions of seven strains of Brucella melitensis,Brucella abortus,and Brucella suis(denoted A,B,C,D,E,F,G)by using 30%glycerol brain heart infusion medium,then conducted stability testing.For accelerated stability tests,the bacterial solution was placed in a 37℃chamber for 1,2,3,or 4 weeks continuously,and the Brucella survival number at each time point was measured.For freeze-thaw stability testing,the bacterial solutions were frozen at-80℃,transferred to room temperature for 4 hours,and melted in one freeze-thaw cycle,and the Brucella survival number was measured for 2,4,6,and 8 freeze-thaw times.For long-term stability testing,the bacterial solutions were stored at-80℃,and the Brucella survival number was measured at various time points during the first,third,sixth,and twelfth months of storage.A line graph was plotted to demonstrate the changes in Brucella survival number at each time point in the three stability tests.IBM SPSS Statistics 22 software was used to assess statistically significant differences in Brucella survival number at each time point and the initial Brucella survival number.Accelerated stability tests showed that the Bru-cella survival number of the seven strains significantly decreased,and a statistically significant difference in Brucella survival number was observed after 1 week.In freeze-thaw stability tests,with increasing freeze-thaw times,the Brucella survival number of the seven strains showed a decreasing trend.Strains C and F showed statistically significant differences with respect to the initial Brucella sur-vival number after two times of freeze-thaw;strains A and D showed statistically significant differences with respect to the initial Bru-cella survival number after four times of freeze-thaw;strain G showed statistically significant differences with respect to the initial Bru-cella survival number after six times of freeze-thaw;and strain E showed statistically significant differences with respect to the initial Brucella survival number after eight times of freeze-thaw.The data for strain B showed abnormalities.In long-term stability testing,the Brucella survival number for strains C and F after storage for 1 month showed a statistically significant difference with respect to the initial bacterial solution.No statistically significant difference in Brucella survival number was observed for strains B and E after stor-age for 1,3,6,or 12 months.The data for strains A,D,and G showed abnormalities.The sterile 30%glycerol brain heart infusion medium maintained the Brucella survival number in an-80℃environment,and freezing and thawing multiple times should be avoided.Compared with the traditional freeze-drying method,this method is not only easy to perform but also avoids potential bio-safety hazards,and provides a reliable Brucella culture preservation scheme for researchers in related fields.

Distal transradial access(dTRA)was first applied in coronary heart disease intervention by Kiemeneij in 2017,dTRA has become an important technological advancement of coronary interventional therapy.This approach,performed by puncturing the distal radial artery within the anatomical snuffbox or Hegu point,significantly reduces the incidence of radial artery occlusion compared with conventional transradial access and shortens compression time to 2-3 hours after procedure.Clinical evidence confirms the efficacy of dTRA in facilitating complex percutaneous coronary interventions,including stenting of left main coronary artery bifurcation lesions and recanalization of chronic total occlusions.Its application has progressively expanded to neurointerventional procedures(cerebral angiography),tumor embolization(transarterial chemoembolization for hepatocellular carcinoma),and peripheral vascular interventions.Despite increasing clinical adoption,dTRA still faces challenges,including a long learning curve and relatively lower initial puncture success rates.Combined with the current paucity of robust evidence-based data,the viability of dTRA as a routine interventional access route remains a subject of debate.This article systematically reviews the anatomical landmarks,clinical advantages,limitations,and multidisciplinary applications of dTRA.This article aims to provide practical guidance for interventionalists and promote the standardization of this technique in daily clinical practice.

Objective To evaluate the value of imaging quantification parameters in artificial intelligence (AI) assisted diagnosis systems in clinical decision-making for lung nodules≤2 cm and the diagnostic efficacy of AI. Methods Lung nodule patients admitted to Affiliated Zhongshan Hospital of Dalian University from 2020 to 2023 were included. Imaging parameters of lung nodules were extracted using AI assisted diagnosis systems. Multifactor analysis was used to screen predictors for distinguishing benign and malignant nodules and high-risk predictors for recurrent invasive adenocarcinoma, and a diagnostic model was established and its performance evaluated. The diagnostic efficacy of the AI system was judged according to pathological results. Results A total of 594 patients with lung nodules were included, including 202 males and 392 females, with an average age of (58.75±11.55) years. Volume, average CT value, and 3D maximum diameter of non-solid nodules were independent predictors of malignant nodules, with thresholds of 287.4 mm3, −491 HU, and 12.0 mm, respectively. The area under the curve (AUC) for diagnostic efficacy was ranked from high to low as combined model (0.802), volume (0.783), average CT value (0.749), and 3D maximum diameter (0.714). The average CT value and 3D long diameter of solid nodules were independent predictors of malignant nodules, with thresholds of −81 HU and 17.5 mm, respectively, and AUC values of 0.874 and 0.686, respectively, with the combined prediction AUC of 0.957. The mass of cystic nodules was an independent predictor of malignancy when the mass>180.7 mg. Independent predictors of high recurrence risk of invasive adenocarcinoma in non-solid nodules were consolidation-tumor ratio (CTR), average CT value, 3D long diameter, and volume, with thresholds of 0.14, −386 HU, 15.6 mm, and 1018.9 mm3, respectively, and diagnostic efficacy was ranked from high to low as combined model (0.788), 3D long diameter (0.735), volume (0.725), average CT value (0.720), and CTR (0.697). The accuracy of AI in predicting benign and malignant target nodules was 87.4%, with positive predictive value of 96.6% and negative predictive value of 58.9%. Conclusion In clinical decision-making for lung nodules ≤2 cm, AI assisted diagnosis systems have high application value.

Current epilepsy prediction methods are not effective in characterizing the multi-domain features of complex long-term electroencephalogram (EEG) data, leading to suboptimal prediction performance. Therefore, this paper proposes a novel multi-scale sparse adaptive convolutional network based on multi-head attention mechanism (MS-SACN-MM) model to effectively characterize the multi-domain features. The model first preprocesses the EEG data, constructs multiple convolutional layers to effectively avoid information overload, and uses a multi-layer perceptron and multi-head attention mechanism to focus the network on critical pre-seizure features. Then, it adopts a focal loss training strategy to alleviate class imbalance and enhance the model's robustness. Experimental results show that on the publicly created dataset (CHB-MIT) by MIT and Boston Children's Hospital, the MS-SACN-MM model achieves a maximum accuracy of 0.999 for seizure prediction 10 ~ 15 minutes in advance. This demonstrates good predictive performance and holds significant importance for early intervention and intelligent clinical management of epilepsy patients.
Humans ; Electroencephalography/methods* ; Epilepsy/physiopathology* ; Neural Networks, Computer ; Seizures/physiopathology* ; Signal Processing, Computer-Assisted ; Algorithms

Objective:To investigate the clinical, imaging and neuropathological characteristics of neuronal intranuclear inclusion disease (NIID) with symptoms of the central nervous system, and to improve the diagnosis and treatments of NIID.Methods:The clinical data of 7 patients with NIID diagnosed by brain biopsy in Xuanwu Hospital, Capital Medical University, Beijing, China from February 2009 to December 2024 were collected. The characteristics of clinical manifestations, imaging, and histology on brain biopsy were retrospectively analyzed.Results:Among the 7 patients, 5 were male and 2 were female. Their ages ranged from 44 to 70 years, median 56 (52, 65) years. Patients were classified into three types of tumor, stroke and encephalitis according to the onset symptoms, imaging manifestations and pathological changes. The chief complaint of the 5 patients was headache, while 4 patients had paroxysmal convulsions, 3 had speech disorders, 2 had abnormal mental behaviors, 2 had memory decline, and 1 had fever accompanied by consciousness disorders. Diffusion-weighted magnetic resonance imaging of the head showed the "ribbon sign" at the junction of the cortex and medulla in 2 cases. Most of the patients had white matter lesions, gyrus swelling and cerebral atrophy. Occasionally gyrus-like enhancement was observed. Brain biopsy reveals the histological changes that matched those on images and initial symptoms. There were proliferation of oligodendrocytes and astrocytes in the white matter, leukoaraiosis and edema, cortical disintegration and lamellar necrosis, as well as infiltration of lymphocytes and microglia, etc. However, the characteristic changes were eosinophilic hyaline inclusions in the nuclei of neurons and astrocytes. Immunohistochemical staining of p62 and ubiquitin showed homogeneous staining in round or ring-shaped nuclei.Conclusions:The clinical manifestations of NIID are highly variable, and a correct diagnosis of NIID requires careful integration of clinical, imaging and histopathologic data. For patients with a high suspicion of NIID, immunohistochemical staining of p62 and ubiquitin is diagnostically valuable.

Objective:To discuss the effect of Yes-associated protein(YAP)silencing on the proliferation,migration,and invasion capabilities of the human cervical cancer(CC)SiHa cells.Methods:The human CC SiHa cells were cultured in vitro,and the lentiviral YAP shRNA was transfected into the SiHa cells to establish stably transfected YAP-shRNA experimental group(sh-YAP group)and empty plasmid control group(control group).Western blotting method was used to detect the silencing effect of YAP;immunofluorescence method was used to detect the microfilament number and morphology of actin filaments(F-actin)in the cells in both groups;CCK-8 method was used to detect the survival rates of the cells in two groups;Transwell chamber assay and wound healing assay were used to detect the numbers of migration and invasion cells and scratch healing rates of the cells in two groups;Western blotting method was used to detect the expression levels of epithelial-mesenchymal transition(EMT)markers(E-cadherin and Snail),DNA damage repair-related proteins(γ-H2AX),and apoptosis-related proteins[c-MYC and B-cell lymphoma-2(Bcl-2)]in the cells in two groups.Results:The results of lentiviral YAP shRNA transfection into SiHa cells showed that the expression level of YAP protein in the SiHa cells was significantly decreased(P＜0.05).The immunofluorescence results showed that after YAP silencing,the F-actin in SiHa cells was sparse and regularly arranged,with a reduced number of cells and a shriveled appearance.The CCK-8 results showed that compared with control group,the survival rate of the SiHa cells in sh-YAP group was significantly decreased cultured for 24 and 48 h(P＜0.01).The results of Transwell chamber assay and the wound healing assay showed that compared with control group,the numbers of migration and invasion SiHa cells in sh-YAP group were significantly decreased(P＜0.01),and the cell scratch healing rates were signifiantly decreased(P＜0.05).The Western blotting results showed that compared with control group,the expression level of E-cadherin protein in the cells in sh-YAP group was increased(P＜0.05),and the expression levels of c-MYC,Bcl-2,and γ-H2AX proteins were decreased(P＜0.05 or P＜0.01).Conclusion:YAP gene silencing leads to the depolymerization of F-actin in the human CC SiHa cells and regulates the apoptosis and DNA damage repair,potentially reversing the EMT process,thereby inhibiting the proliferation and migration of the tumor cells.

Yttrium-90(90Y)selective internal radiation therapy(SIRT)is an emerging modality for the treatment of hepatocellular carcinoma(HCC),leveraging the nuclide 90Y to deliver targeted radiation therapy.90Y has a long half-life and can be used to selectively ablate tumor cells by high-energy beta rays.It has high biological effectiveness and robust local control capabilities.In recent years,with the continuous advancement of basic and clinical research,the application of 90Y-SIRT in the conversion treatment of unresectable HCC(uHCC)has made significant progress.However,challenges remain in the clinical application of 90Y-SIRT,including how to improve the efficacy of conversion therapy and how to optimize therapy regimens.This review aims to summarize the research progress of90Y-SIRT in the conversion therapy of uHCC.