BACKGROUND:Traditional methods of urinary tract reconstruction are limited by donor scarcity,high complication rates,and suboptimal functional recovery.Tissue engineering strategies offer new directions in this field.Since the urinary tract is mainly composed of muscle tissue,the key is to find suitable seed cells and efficiently induce them to differentiate into smooth muscle cells.Comparative studies on the efficacy of different smooth muscle cell induction regimens are still lacking. OBJECTIVE:To isolate,culture,and identify human urine-derived stem cells,and to compare the effects of two different induction protocols. METHODS:Human urine-derived stem cells were isolated from urine samples of 11 healthy adult volunteers by multiple centrifugations.Surface markers were identified by flow cytometry.The multi-directional differentiation potential of human urine-derived stem cells was verified through osteogenic and adipogenic differentiation.Differentiation was induced by transforming growth factor-β1 or transforming growth factor-β1 combined with platelet derived growth factor for 14 days.Immunofluorescence staining and western blot assay were employed to compare the expression differences of smooth muscle-specific proteins(α-SMA and SM22). RESULTS AND CONCLUSION:(1)Urine-derived stem cells were successfully isolated from the eight urine samples of healthy people.These cells exhibit a"rice grain"-like morphology and possess a robust proliferative capacity.(2)Urine-derived stem cells exhibited high expression of mesenchymal stem cell surface markers(CD73,CD90,and CD44)and extremely low expression of hematopoietic stem cell surface markers(CD34 and CD45).These cells did not express CD19,CD105,and HLA-DR.(3)After osteogenic and adipogenic differentiation,the formation of calcium nodules and lipid droplets was observed,with positive staining results from Alizarin Red S and Oil Red O staining.(4)After 14 days of smooth muscle induction culture,immunofluorescence staining revealed that the smooth muscle differentiation rate of urine-derived stem cells treated with a combination of transforming growth factor-β1 and platelet derived growth factor was significantly higher compared to those treated with transforming growth factor-β1 alone(P<0.005).(5)After 14 days of smooth muscle induction culture,western blot assay further demonstrated that the expression levels of α-SMA and SM22 in the transforming growth factor-β1/platelet derived growth factor group were significantly elevated compared to those in the transforming growth factor-β1 only group(P<0.005).These findings confirm that urine-derived stem cells can be non-invasively isolated using multiple rounds of centrifugation.Compared with transforming growth factor-β1 alone,the combination of transforming growth factor-β1 and platelet derived growth factor can improve the efficiency of inducing urine-derived stem cells to differentiate into smooth muscle cells.

OBJECTIVE To investigate the targeting effect of folic acid-modified crebanine nanoparticles （FA-Cre@PEG- PLGA NPs， hereinafter referred to as “NPs”） combined with ultrasound irradiation on M109 cells in vitro and in vivo after administration， and explore the anti-tumor mechanism. METHODS CCK-8 assay was used to detect the inhibitory effect of NPs combined with ultrasound irradiation on the proliferation of M109 cells， and the best ultrasound time was selected. Using human lung cancer A549 cells as a control， the targeting of NPs combined with ultrasound irradiation to M109 cells was evaluated by free folic acid blocking assay and cell uptake assay. The effects of NPs combined with ultrasound irradiation on the migration， invasion， apoptosis， cell cycle and reactive oxygen species （ROS） levels of M109 cells were detected by cell scratch test， Transwell chamber test and flow cytometry at 1 h after 958401536@qq.com administration； the changes of mitochondrial membrane potential （MMP） were observed by fluorescence inverted microscope. A mouse subcutaneous tumor model of M109 cells was constructed， and the in vivo tumor targeting of NPs combined with ultrasound irradiation was investigated by small animal in vivo imaging technology. RESULTS NPs combined with ultrasound irradiation could significantly inhibit the proliferation of M109 cells， and the optimal ultrasound time was 1 h after administration. The free folic acid could antagonize the inhibitory effect of NPs on the proliferation of M109 cells， and combined with ultrasound irradiation could partially reverse this antagonism. Compared with A549 cells， the uptake rate of NPs in M109 cells was significantly higher （P＜0.01）， and ultrasound irradiation could promote cellular uptake. NPs combined with ultrasound irradiation could inhibit the migration and invasion of M109 cells and block the cell cycle in the G0/G1 and G2/M phases. Compared with control group， the apoptosis rate of M109 cells and ROS level were increased significantly （P＜0.01）， while the MMP decreased significantly （P＜0.01） in the different concentration （100， 200， 300 μg/mL） groups of M109 cells. Compared with the mice in non-ultrasound group， the fluorescence intensity and tumor-targeting index of the tumor site in the 0 h ultrasound group were significantly enhanced （P＜0.05 or P＜0.01）. CONCLUSIONS NPs combined with ultrasound irradiation have a strong targeting effect on M109 cells in vitro and in vivo， the anti-tumor mechanism includes inhibiting cell migration and invasion， blocking cell cycle， and inducing apoptosis.

This study aims to reveal the effect and mechanism of Gentianella turkestanorum total extract(GTI) in ameliorating non-alcoholic steatohepatitis(NASH). UPLC-Q-TOF-MS was employed to identify the chemical components in GTI. SwissTarget-Prediction, GeneCards, OMIM, and TTD were utilized to screen the targets of GTI components and NASH. The common targets shared by GTI components and NASH were filtered through the STRING database and Cytoscape 3.9.0 to identify core targets, followed by GO and KEGG enrichment analysis. AutoDock was used for molecular docking of key components with core targets. A mouse model of NASH was established with a methionine-choline-deficient high-fat diet. A 4-week drug intervention was conducted, during which mouse weight was monitored, and the liver-to-brain ratio was measured at the end. Hematoxylin-eosin staining, Sirius red staining, and oil red O staining were employed to observe the pathological changes in the liver tissue. The levels of various biomarkers, including aspartate aminotransferase(AST), alanine aminotransferase(ALT), hydroxyproline(HYP), total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione(GSH), in the serum and liver tissue were determined. RT-qPCR was conducted to measure the mRNA levels of interleukin 1β(IL-1β), interleukin 6(IL-6), tumor necrosis factor α(TNF-α), collagen type I α1 chain(COL1A1), and α-smooth muscle actin(α-SMA). Western blotting was conducted to determine the protein levels of IL-1β, IL-6, TNF-α, and potential drug targets identified through network pharmacology. UPLC-Q-TOF/MS identified 581 chemical components of GTI, and 534 targets of GTI and 1 157 targets of NASH were screened out. The topological analysis of the common targets shared by GTI and NASH identified core targets such as IL-1β, IL-6, protein kinase B(AKT), TNF, and peroxisome proliferator activated receptor gamma(PPARG). GO and KEGG analyses indicated that the ameliorating effect of GTI on NASH was related to inflammatory responses and the phosphoinositide 3-kinase(PI3K)/AKT pathway. The staining results demonstrated that GTI ameliorated hepatocyte vacuolation, swelling, ballooning, and lipid accumulation in NASH mice. Compared with the model group, high doses of GTI reduced the AST, ALT, HYP, TC, and TG levels(P<0.01) while increasing the HDL-C, SOD, and GSH levels(P<0.01). RT-qPCR results showed that GTI down-regulated the mRNA levels of IL-1β, IL-6, TNF-α, COL1A1, and α-SMA(P<0.01). Western blot results indicated that GTI down-regulated the protein levels of IL-1β, IL-6, TNF-α, phosphorylated PI3K(p-PI3K), phosphorylated AKT(p-AKT), phosphorylated inhibitor of nuclear factor kappa B alpha(p-IκBα), and nuclear factor kappa B(NF-κB)(P<0.01). In summary, GTI ameliorates inflammation, dyslipidemia, and oxidative stress associated with NASH by regulating the PI3K/AKT/NF-κB signaling pathway.
Animals ; Non-alcoholic Fatty Liver Disease/genetics* ; Mice ; Network Pharmacology ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Chromatography, High Pressure Liquid ; Liver/metabolism* ; Mice, Inbred C57BL ; Humans ; Mass Spectrometry ; Tumor Necrosis Factor-alpha/metabolism* ; Disease Models, Animal ; Molecular Docking Simulation

Objective·To construct and verify the mouse model that can mimic the vitamin A deficiency(VAD)-like craniofacial skeletal deformity and do not cause embryonic death.Methods·Based on the Cre-LoxP system,the OsxCre;Rosa26dn/dn mice expressing osteoblast-specific dominant-negative retinoid acid receptor α(dnRARα)mutation were obtained by hybridization through OsxCre and Rosa26dnRARa/ddnRARa mice,to achieve the conditional inhibition of retinoic acid signaling to simulate VAD disease.Femur bone mesenchymal stem cells(BMSCs)and parietal bone cells of OsxCre;Rosa26dn/dn mice and their control littermates were isolated and underwent osteogenic induction,to assess the expression of retinoid acid receptor α(RARα)protein through Western blotting.Osteoblasts induced from parietal bone cells of OsxCre;Rosa26dn/dn mice and their control littermates were isolated and the effect of retinoic acid signaling inhibition was verified through dual luciferase gene reporter assay.Meanwhile,Ad-eGFP or Ad-Cre adenovirus-infected femur BMSCs and parietal bone cells of Rosa26dn/dnmice underwent osteogenic induction to assess the expression of dominant-negative mutant protein and the inhibition of the retinoic acid signaling pathway in vitro by Western blotting and dual luciferase gene reporter assay.Moreover,the skulls of 6-week-old OsxCre;Rosa26dn/dn mice were collected,and Micro-CT scanning and three-dimensional(3D)reconstruction were performed to verify the craniofacial skeletal deformities of the mouse model.Results·Western blotting results demonstrated that the level of RARα protein increased in the femur and parietal osteoblasts of OsxCre;Rosa26dn/dn mice compared to that of their control littermates,and also increased in the Ad-Cre-infected femur and parietal osteoblasts of Rosa26dn/dn mice compared to that in the Ad-eGFP-infected group(P＜0.05).Dualluciferase gene reporter assay results indicated that the activity of retinoid acid response element(RARE)was inhibited in the osteoblasts of OsxCre;Rosa26dn/dn mice compared to their control littermates,and was also inhibited in the Ad-Cre-infected group compared to the Ad-eGFP-infected group(P＜0.05).Micro-CT and 3D reconstruction suggested that the skull of 6-week-old OsxCre;Rosa26dn/dn mice exhibited VAD-like craniofacial skeletal deformities,including smaller size of the skull and osteogenesis imperfecta compared to their control littermates.Conclusion·An osteoblast-specific dnRARα expressing mouse model that can mimic VAD-like craniofacial skeletal deformity is successfully constructed,therefore providing a new model for exploring the pathogenesis and therapeutic targets of VAD-like craniofacial skeletal deformity in the future.

Purpose::Vibrio vulnificus ( V. Vulnificus) infection is characterized by rapid onset, aggressive progression, and challenging treatment. Bacterial resistance poses a significant challenge for clinical anti-infection treatment and is thus the subject of research. Enhancing host infection tolerance represents a novel infection prevention strategy to improve patient survival. Our team initially identified cytochrome P4501A1 (CYP1A1) as an important target owing to its negative modulation of the body's infection tolerance. This study explored the superior effects of the CYP1A1 inhibitor bergamottin compared to antibiotic combination therapy on the survival of mice infected with multidrug-resistant V. Vulnificus and the protection of their vital organs. Methods::An increasing concentration gradient method was used to induce multidrug-resistant V. Vulnificus development. We established a lethal infection model in C57BL/6J male mice and evaluated the effect of bergamottin on mouse survival. A mild infection model was established in C57BL/6J male mice, and the serum levels of creatinine, urea nitrogen, aspartate aminotransferase, and alanine aminotransferase were determined using enzyme-linked immunosorbent assay to evaluate the effect of bergamottin on liver and kidney function. The morphological changes induced in the presence of bergamottin in mouse organs were evaluated by hematoxylin and eosin staining of liver and kidney tissues. The bacterial growth curve and organ load determination were used to evaluate whether bergamottin has a direct antibacterial effect on multidrug-resistant V. Vulnificus. Quantification of inflammatory factors in serum by enzyme-linked immunosorbent assay and the expression levels of inflammatory factors in liver and kidney tissues by real-time quantitative polymerase chain reaction were performed to evaluate the effect of bergamottin on inflammatory factor levels. Western blot analysis of IκBα, phosphorylated IκBα, p65, and phosphorylated p65 protein expression in liver and kidney tissues and in human hepatocellular carcinomas-2 and human kidney-2 cell lines was used to evaluate the effect of bergamottin on the nuclear factor kappa-B signaling pathway. One-way ANOVA and Kaplan-Meier analysis were used for statistical analysis. Results::In mice infected with multidrug-resistant V. Vulnificus, bergamottin prolonged survival ( p = 0.014), reduced the serum creatinine ( p = 0.002), urea nitrogen ( p = 0.030), aspartate aminotransferase ( p = 0.029), and alanine aminotransferase ( p = 0.003) levels, and protected the cellular morphology of liver and kidney tissues. Bergamottin inhibited interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α expression in serum (IL-1β: p = 0.010, IL-6: p = 0.029, TNF-α: p = 0.025) and inhibited the protein expression of the inflammatory factors IL-1β, IL-6, TNF-α in liver (IL-1β: p = 0.010, IL-6: p = 0.011, TNF-α: p = 0.037) and kidney (IL-1β: p = 0.016, IL-6: p = 0.011, TNF-α: p = 0.008) tissues. Bergamottin did not affect the proliferation of multidrug-resistant V. Vulnificus or the bacterial load in the mouse peritoneal lavage fluid ( p = 0.225), liver ( p = 0.186), or kidney ( p = 0.637). Conclusion::Bergamottin enhances the tolerance of mice to multidrug-resistant V. Vulnificus infection. This study can serve as a reference and guide the development of novel clinical treatment strategies for V. Vulnificus.

Objective To investigate the effect of parent-child alienation on depression in surface ship officers and soldiers based on the theory of"diathesis-stress",and the mediating role of resilience between parent-child alienation and depression in them.Methods A cross-sectional study was conducted on 599 officers and soldiers from a surface ship unit.The participants were surveyed with inventory of alienation toward parents,connor-davidson resilience scale and patient health questionnaire-9 to obtain and analyze their demographic-military characteristics of their depression scores.The participants with depression scores ≥5 were recruited as the subjects,and Spearman correlation analysis was used to explore the correlation among parent-child alienation,resilience and depression.On the basis of hierarchical regression analysis,AMOS software was used to establish a structural equation modelling of intermediary effects.Results The depression score was 1(0,4)in the participants,and the depression scores of those with service length ≥11 years were comparatively higher than those with shorter length.Our results indicated that parent-child alienation was positive correlated with depression(r=0.451,P＜0.001),while resilience was negatively correlated with depression and parent-child alienation(r=-0.412,-0.407,P＜0.001).Regression analysis revealed that parent-child alienation had a direct positive predictive value for depression(β=0.574,P＜0.001),and resilience showed a negative predictive value for depression(β=-0.211,P＜0.01).Model analysis displayed that resilience had a significant mediating role in the effect of parent-child alienation on depression among these surface ship officers and soldiers,with an effect value of 0.088,and accounting for 15.86％of the total effect.Conclusion Parent-child alienation has a significant influence on depression among surface ship officers and soldiers,with resilience playing a partial mediating role.

Objective To observe the clinical effect of Tongyuan Acupuncture combined with Modified Linggui Zhugan Decoction in the treatment of polycystic ovary syndrome(PCOS)with spleen deficiency and phlegm-damp syndrome.Methods A total of 86 patients with PCOS of spleen deficiency and phlegm-damp type were randomly divided into observation group and control group,43 cases in each group.The control group was treated with Metformin Hydrochloride Tablets combined with Ethinylestradiol and Cyproterone Acetate Tablets.The observation group was treated with Tongyuan Acupuncture combined with Modified Linggui Zhugan Decoction on the basis of the treatment of the control group.Both groups were treated for three consecutive months.After three months of treatment,the clinical efficacy of the two groups was evaluated.The changes of sex hormone indexes estradiol(E2),testosterone(T),luteinizing hormone(LH),follicle stimulating hormone(FSH)and prolactin(PRL)expression levels were observed before and after treatment in the two groups.The changes of glucose metabolism levels including insulin resistance index(HOMA-IR),fasting insulin(FINS)and fasting blood glucose(FPG)expression levels were compared before and after treatment between the two groups.Results(1)The total effective rate was 90.70%(39/43)in the observation group and 74.42%(32/43)in the control group.The curative effect of the observation group was superior to that of the control group,and the difference was statistically significant(P＜0.05).(2)After treatment,HOMA-IR,FINS and FPG in the two groups were significantly improved(P＜0.05),and the improvement of HOMA-IR,FINS and FPG in the observation group was superior to that in the control group,the difference being significant(P＜0.05).(3)After treatment,the levels of E2,T,LH,FSH and PRL in the two groups were significantly improved(P＜0.05),and the levels of E2,T,LH,FSH and PRL in the observation group were significantly superior to those in the control group,the differences were statistically significant(P＜0.05).Conclusion Tongyuan Acupuncture combined with Modified Linggui Zhugan Decoction in the treatment of spleen deficiency and phlegm-damp type of PCOS can effectively improve the sex hormone level of patients and regulate the glucose metabolism of patients.The clinical effect is significant.

Objective To investigate the occurrence of adverse events of lurasidone in the U.S.Food and Drug Administration Adverse Event Reporting System(FAERS)database by using Open Vigil FDA2.1,to enrich the experience and provide the basis for the clinical use of the drug in China.Methods Using Open Vigil FDA2.1,adverse event data were extracted from the FAERS database for a total of 51 quarters from the 4th quarter of 2010 to the 3rd quarter of 2023,and the ratio of reporting ratio(ROR)method and the proportional reporting ratio(PRR)method were used for data mining and analysis.Results A total of 32 728 adverse event reports with lurasidone as the first suspected drug was obtained,with a larger proportion of females(54.26％)and occurring mostly in adults(18 to 59 years).After the screening,326 preferred term(PT)signals were obtained,involving 20 system-organ classifications(injury,poisoning and procedural complications,general disorders and administration site conditions,psychiatric disorders,etc.).Among them,PTs with the higher frequency of occurrence included off label use,feeling abnormal,crying,anxiety,depression,insomnia,etc.PTs with stronger signal strength included activation syndrome,mania,tongue movement disturbance,hypoprolactinaemia,akathisia,etc.Multiple new suspected adverse drug reactions were unearthed,including hypoprolactinaemia,emotional poverty,stiff tongue,etc.Conclusion Lurasidone has a favorable safety profile,and women need to closely monitor prolactin levels when taking this medication.The drug is relatively safe for use in pregnant,puerperal and perinatal women and patients with poor metabolic function.Hypoprolactinaemia and restless leg syndrome are new rare suspected adverse events with lurasidone.

Dendrobium officinale is a Chinese herbal medicine with the same origin as medicine and food,and it is commonly used in making soup and tonic diet,etc.Dendrobium officinale and its compound preparations are widely used in clinical practice.This article used keywords such as"Dendrobium officinale"and"clinical"to search for relevant literature on clinical research on Dendrobium officinale in CNKI,Wanfang,VIP,PubMed and other databases over the past 10 years.It is found that Dendrobium officinale and its compound preparations were mainly used in the treatment of internal medicine(such as chronic gastritis,diabetes,bronchitis,hypertension,coronary heart disease,etc.),surgery,gynecology,pediatrics,ophthalmology and other diseases,and had good clinical effects.It is a Chinese herbal medicine with the development value.This article provides a comprehensive and systematic summary and analysis of the clinical application of Dendrobium officinale,aiming to provide reference for its rational clinical application and further development and research.

Objective:To evaluate the clinical efficacy of pushing reduction with our self-designed spinal fracture reduction device in the treatment of A3N0/1 thoracolumbar fractures.Methods:A retrospective study was conducted to analyze the medical records of 53 patients who had undergone surgery for thoracolumbar vertebrae fracture at Department of Minimally Invasive Spine Surgery, Zhengzhou Orthopedic Hospital from January 2019 to January 2022. All patients were treated by internal fixation via the Wiltse approach and bone grafting through the pedicle of the injured vertebrae. Clinical data: 35 males and 18 females; age: (37.8±10.2) years; injured segments: 23 cases at the thoracic spine and 30 cases at the lumbar spine; time from injury to surgery: (3.3±1.5) days. According to whether our self-designed spinal fracture reduction device was used or not, the patients were assigned into group A (23 cases) in which the injured vertebrae were pushed and reduced using our novel spinal fracture reduction device after vertebral distraction reduction by the pedicle screw and group B (30 cases) in which the injured vertebrae were distracted and reduced using the pedicle screw alone. The operation time, intraoperative blood loss and complications were compared between the 2 groups. The anterior vertebral body height ratio (AVBHr), middle vertebral body height ratio (MVBHr), posterior vertebral body height ratio (PVBHr), Cobb angle of the injured vertebra, visual analogue scale (VAS) and Oswestry disability index (ODI) at preoperation, postoperative 3 and 6 months, and the last follow-up were compared between the 2 groups.Results:There was no statistically significant difference in the preoperative general data between the 2 groups, indicating comparability ( P>0.05). All patients were followed up for (16.3±5.9) months. All incisions healed at one stage postoperatively without any related complications. The operation time in group A was significantly longer than that in group B [(115.1±16.6) min. versus (101.0±11.5) min.], the intraoperative blood loss in group A was significantly greater than that in group B [(136.5±17.0) mL versus (121.6±19.8) mL], the MVBHr at postoperative 3 months in group A (93.9%±4.0%) was significantly better than that in group B (83.3%±7.6%), and the MVBHr, AVBHr, Cobb angle, VAS, and ODI at the last follow-up in group A [86.6%±5.5%, 89.8%±4.1%, 4°(4°, 6°), 1 (0, 1) point, and 4.7%±2.0%] were significantly better than those in group B [78.0% (74.0%, 79.0%), 84.5%±4.9%, 12.2°±3.3°, 2 (1, 3) points, and 7.3%±2.7%] (all P<0.05). However, there was no statistically significant difference in PVBHr between the 2 groups at postoperative 3 months or at the last follow-up ( P>0.05). Conclusion:In the treatment of A3N0/1 thoracolumbar fractures, pushing reduction with our self-designed spinal fracture reduction device can directly and effectively reduce the fracture zone of the injured vertebra, which is conducive to maintaining postoperative vertebral reduction, reducing vertebral height loss and kyphotic deformity at a later stage, relieving lumbar pain and improving lumbar spine function.