1.Ectomesenchymal stem cells-derived extracellular vesicles promote neuronal axonal elongation
Haitao SUN ; Chunpeng REN ; Yongtao YANG ; Yonghui HUANG ; Rujie QIN ; Zhen LI
Chinese Journal of Tissue Engineering Research 2025;29(23):4924-4930
BACKGROUND:The occurrence of neuronal axonal injury can result in neurological dysfunction,and the facilitation of axonal elongation is anticipated to play a pivotal role in the treatment of diseases affecting the nervous system.OBJECTIVE:To investigate whether ectomesenchymal stem cells-derived extracellular vesicles can promote neuronal axonal elongation.METHODS:(1)Ectomesenchymal stem cells were obtained from nasal mucosa using the tissue adherence method,and the specific markers of were identified through immunofluorescence.Ectomesenchymal stem cells-derived extracellular vesicles were acquired via ultracentrifugation and identified.(2)Ectomesenchymal stem cells-derived extracellular vesicles(0,0.5,1.0,1.5 mg/mL)were incubated with PC12 cells for 72 hours.The cytotoxicity and proliferation of ectomesenchymal stem cells-derived extracellular vesicles on PC12 cells were assessed using the CCK-8 assay.(3)Ectomesenchymal stem cells-derived extracellular vesicles(1.0 mg/mL)were incubated with PC12 cells or neurons for 72 hours.The changes in axon length were observed using microscopic analysis.The expression levels of axon-related markers β3-tubulin(early stage),growth associated protein 43(middle stage),and neurofilament 200(mature stage)were analyzed through real-time fluorescence quantitative PCR and Western blotting.These investigations aimed to explore the potential of ectomesenchymal stem cells-derived extracellular vesicles in promoting neurite elongation within PC12 cells or neurons.RESULTS AND CONCLUSION:(1)The majority of the acquired ectomesenchymal stem cells exhibited a spindle-shaped morphology,while a minority displayed irregular shapes,and demonstrated high expression levels of mesenchymal stem cell-specific markers Nestin,CD44,and Vimentin.The obtained ectomesenchymal stem cells-derived extracellular vesicles fulfilled the biological criteria for extracellular vesicles.(2)Within the detected protein concentration range of 0.5 to 1.5 mg/mL,the proliferation of PC12 cells was promoted by ectomesenchymal stem cells-derived extracellular vesicles,and this effect was further enhanced with increasing concentrations.(3)Ectomesenchymal stem cells-derived extracellular vesicles increased the length of axons in PC12 cells and neurons and the expression of axon-related markers β3-tubulin,growth associated protein 43,and neurofilament 200.Above findings suggest that ectomesenchymal stem cells-derived extracellular vesicles have the potential to enhance neuronal axonal elongation.
2."Two-point and two-line method" in design of free perforator flap of medial sural artery: clinical application and clinical significance
Yan ZHANG ; Yucheng LIU ; Yang CAO ; Haibo WU ; Yongtao HUANG ; Chengpeng YANG ; Fengwen SUN ; Qinfeng GAO ; Jihui JU ; Guangzhe JIN
Chinese Journal of Microsurgery 2025;48(1):55-59
Objective:To study the feasibility of design and harvest of free medial sural artery perforator flap with the "two-point and two-line method".Methods:From September 2022 to June 2023, Suzhou Ruihua Orthopaedic Hospital implemented the "two-point and two-line method" to guide preoperative perforator positioning and flap design. Thirty medial sural artery perforator flaps were successfully harvested with the method, and 21 wounds of hand and 9 of foot and ankle were reconstructed with the flaps. The size of soft tissue defects were 2.5 cm×2.5 cm-7.0 cm×14.5 cm, and the flaps size were 3.0 cm×3.0 cm-7.5 cm×15.0 cm. All donor sites were directly closed or by skin grafting. All patients were entered in 6-15 months of postoperative outpatient follow-up, and the recovery of donor and recipient sites was assessed by the comprehensive evaluation scales. The sensory function of the flaps was evaluated using the sensory function evaluation standard of British Medical Research Council (BMRC).Results:All perforators were successfully located with 47 perforators in total, and all of them were musculocutaneous perforator. It was found that there was 1 perforator in 14 flaps, 2 perforators in 15 flaps and 3 perforators in 1 flap. All 30 flaps survived after surgery, beside 2 flaps that had arterial insufficiency but survived successfully after surgical exploration. All donor sites healed in one stage. Comprehensive evaluation scale of flap was employed to evaluate the flaps and the scores were 84 points to 96 points with an average score of 92.5 points. The excellent and good grades were achieved in 27 flaps and 3 flaps, respectively, with a combined excellent and good rate at 100%. Sensation of the flaps was evaluated by BMRC with 1 flap of S 1, 17 of S 2 and 12 of S 3. Conclusion:The "two-point and two-line method" has been used in design of the perforator flap of medial sural artery. This method is simple and accurate, and is feasible and ideal in design of flaps before surgery.
3.Ectomesenchymal stem cells-derived extracellular vesicles promote neuronal axonal elongation
Haitao SUN ; Chunpeng REN ; Yongtao YANG ; Yonghui HUANG ; Rujie QIN ; Zhen LI
Chinese Journal of Tissue Engineering Research 2025;29(23):4924-4930
BACKGROUND:The occurrence of neuronal axonal injury can result in neurological dysfunction,and the facilitation of axonal elongation is anticipated to play a pivotal role in the treatment of diseases affecting the nervous system.OBJECTIVE:To investigate whether ectomesenchymal stem cells-derived extracellular vesicles can promote neuronal axonal elongation.METHODS:(1)Ectomesenchymal stem cells were obtained from nasal mucosa using the tissue adherence method,and the specific markers of were identified through immunofluorescence.Ectomesenchymal stem cells-derived extracellular vesicles were acquired via ultracentrifugation and identified.(2)Ectomesenchymal stem cells-derived extracellular vesicles(0,0.5,1.0,1.5 mg/mL)were incubated with PC12 cells for 72 hours.The cytotoxicity and proliferation of ectomesenchymal stem cells-derived extracellular vesicles on PC12 cells were assessed using the CCK-8 assay.(3)Ectomesenchymal stem cells-derived extracellular vesicles(1.0 mg/mL)were incubated with PC12 cells or neurons for 72 hours.The changes in axon length were observed using microscopic analysis.The expression levels of axon-related markers β3-tubulin(early stage),growth associated protein 43(middle stage),and neurofilament 200(mature stage)were analyzed through real-time fluorescence quantitative PCR and Western blotting.These investigations aimed to explore the potential of ectomesenchymal stem cells-derived extracellular vesicles in promoting neurite elongation within PC12 cells or neurons.RESULTS AND CONCLUSION:(1)The majority of the acquired ectomesenchymal stem cells exhibited a spindle-shaped morphology,while a minority displayed irregular shapes,and demonstrated high expression levels of mesenchymal stem cell-specific markers Nestin,CD44,and Vimentin.The obtained ectomesenchymal stem cells-derived extracellular vesicles fulfilled the biological criteria for extracellular vesicles.(2)Within the detected protein concentration range of 0.5 to 1.5 mg/mL,the proliferation of PC12 cells was promoted by ectomesenchymal stem cells-derived extracellular vesicles,and this effect was further enhanced with increasing concentrations.(3)Ectomesenchymal stem cells-derived extracellular vesicles increased the length of axons in PC12 cells and neurons and the expression of axon-related markers β3-tubulin,growth associated protein 43,and neurofilament 200.Above findings suggest that ectomesenchymal stem cells-derived extracellular vesicles have the potential to enhance neuronal axonal elongation.
4."Two-point and two-line method" in design of free perforator flap of medial sural artery: clinical application and clinical significance
Yan ZHANG ; Yucheng LIU ; Yang CAO ; Haibo WU ; Yongtao HUANG ; Chengpeng YANG ; Fengwen SUN ; Qinfeng GAO ; Jihui JU ; Guangzhe JIN
Chinese Journal of Microsurgery 2025;48(1):55-59
Objective:To study the feasibility of design and harvest of free medial sural artery perforator flap with the "two-point and two-line method".Methods:From September 2022 to June 2023, Suzhou Ruihua Orthopaedic Hospital implemented the "two-point and two-line method" to guide preoperative perforator positioning and flap design. Thirty medial sural artery perforator flaps were successfully harvested with the method, and 21 wounds of hand and 9 of foot and ankle were reconstructed with the flaps. The size of soft tissue defects were 2.5 cm×2.5 cm-7.0 cm×14.5 cm, and the flaps size were 3.0 cm×3.0 cm-7.5 cm×15.0 cm. All donor sites were directly closed or by skin grafting. All patients were entered in 6-15 months of postoperative outpatient follow-up, and the recovery of donor and recipient sites was assessed by the comprehensive evaluation scales. The sensory function of the flaps was evaluated using the sensory function evaluation standard of British Medical Research Council (BMRC).Results:All perforators were successfully located with 47 perforators in total, and all of them were musculocutaneous perforator. It was found that there was 1 perforator in 14 flaps, 2 perforators in 15 flaps and 3 perforators in 1 flap. All 30 flaps survived after surgery, beside 2 flaps that had arterial insufficiency but survived successfully after surgical exploration. All donor sites healed in one stage. Comprehensive evaluation scale of flap was employed to evaluate the flaps and the scores were 84 points to 96 points with an average score of 92.5 points. The excellent and good grades were achieved in 27 flaps and 3 flaps, respectively, with a combined excellent and good rate at 100%. Sensation of the flaps was evaluated by BMRC with 1 flap of S 1, 17 of S 2 and 12 of S 3. Conclusion:The "two-point and two-line method" has been used in design of the perforator flap of medial sural artery. This method is simple and accurate, and is feasible and ideal in design of flaps before surgery.
5.Antibiofilm activity of polyethylene glycol-quercetin nanoparticlesloaded gelatin-N,O-carboxymethyl chitosan composite nanogels against Staphylococcus epidermidis
Wanhe LUO ; Yongtao JIANG ; Jinhuan LIU ; Beibei SUN ; Xiuge GAO ; Samah Attia ALGHARIB ; Dawei GUO ; Jie WEI ; Yurong WEI
Journal of Veterinary Science 2024;25(2):e30-
Background:
Biofilms, such as those from Staphylococcus epidermidis, are generally insensitive to traditional antimicrobial agents, making it difficult to inhibit their formation. Although quercetin has excellent antibiofilm effects, its clinical applications are limited by the lack of sustained and targeted release at the site of S. epidermidis infection.
Objectives:
Polyethylene glycol-quercetin nanoparticles (PQ-NPs)-loaded gelatin-N,Ocarboxymethyl chitosan (N,O-CMCS) composite nanogels were prepared and assessed for the on-demand release potential for reducing S. epidermidis biofilm formation.
Methods:
The formation mechanism, physicochemical characterization, and antibiofilm activity of PQ-nanogels against S. epidermidis were studied.
Results:
Physicochemical characterization confirmed that PQ-nanogels had been prepared by the electrostatic interactions between gelatin and N,O-CMCS with sodium tripolyphosphate. The PQ-nanogels exhibited obvious pH and gelatinase-responsive to achieve on-demand release in the micro-environment (pH 5.5 and gelatinase) of S. epidermidis.In addition, PQ-nanogels had excellent antibiofilm activity, and the potential antibiofilm mechanism may enhance its antibiofilm activity by reducing its relative biofilm formation, surface hydrophobicity, exopolysaccharides production, and eDNA production.
Conclusions
This study will guide the development of the dual responsiveness (pH and gelatinase) of nanogels to achieve on-demand release for reducing S. epidermidis biofilm formation.
6.Echocardiographic evaluation on infants with pulmonary atresia and intact ventricular septum:Surgical decision and post operation right ventricle development
Jing YANG ; Wenhong DING ; Qiang WANG ; Bin LI ; Yongtao WU ; Yuekun SUN ; Zhenbo HU
Chinese Journal of Medical Imaging Technology 2024;40(11):1672-1676
Objective To observe value of echocardiographic evaluation on infants with pulmonary atresia and intact ventricular septum(PA/IVS)for surgical decision and post operation right ventricle(RV)development.Methods Forty-six PA/IVS infants who underwent pulmonary valve(PV)annuloplasty(group A,n=25),PV annuloplasty and RV outflow tract reconstruction with/without additional systemic-to-pulmonary artery shunt(group B,n=15),and systemic-to-pulmonary artery shunt and atrial septal enlargement(group C,n=6)were retrospectively enrolled.Status of RV developments were compared among groups before operation as well as 1 and 6 months after operation.Results The presentation age in group B was younger than,while in group C was older than that in group A(both P<0.05).Before operation,tricuspid valve(TV)annulus diameter,TV annulus Z-score and TV/mitral valve(MV)annulus ratio in groups B and C were both smaller than those in group A(all P<0.05),whereas RV/left ventricle(LV)longitudinal diameter ratio in group B was larger and PV annulus Z-score in group C was smaller than those in group A(both P<0.05).Meanwhile,preoperative PV annulus Z-score and RV/LV longitudinal diameter ratio in group B were both larger than those in group C(both P<0.05).One month after operations,TV annulus diameter,TV annulus Z-score and TV/MV annulus ratio in group B and C,as well as PV annulus diameter,PV annulus Z-score and RV/LV longitudinal diameter ratio in group C were all smaller than those in group A(all P<0.05),while PV annulus diameter,PV annulus Z-score and RV/LV longitudinal diameter ratio in group C were all smaller than those in group B(all P<0.05).Six months after operations,no significant difference of TV annulus diameter,PV annulus diameter,PV annulus Z-score,TV/MV annulus ratio nor RV/LV longitudinal diameter ratio was found between group A and B(all P>0.05),but the above indexes in group C were all lower than those in group A and B(all P<0.05).Besides,no significant difference of TV annulus Z-score was found between group A and B(P>0.05),which were lower in group C than in group A(P<0.05).Conclusion Echocardiographic evaluation on PA/IVS infants was helpful to establishment of forward blood flow from RV to pulmonary arteries in time,hence promoting RV development.
7.Application of highly selective arterial indocyanine green angiography in the design of anterolateral thigh free flap
Shi WANG ; Shuai DONG ; Yang CAO ; Guiyang WANG ; Chengpeng YANG ; Fengwen SUN ; Yongtao HUANG ; Liping GUO ; Liang YANG ; Rong ZHOU ; Jihui JU
Chinese Journal of Burns 2024;40(10):948-954
Objective:To introduce the application of highly selective arterial indocyanine green angiography (hereinafter referred to as highly selective arterial angiography) in the design of anterolateral thigh free flap.Methods:This study was a retrospective observational study. From November 2023 to April 2024, 29 patients with wounds in extremities which were repaired by anterolateral thigh free flaps designed under the assistance of highly selective arterial angiography and met the inclusion criteria were admitted to the Department of Hand Surgery and Department of Wound Repair Surgery of Suzhou Ruihua Orthopedic Hospital, including 26 males and 3 females, aged 16 to 71 years. The wound area after debridement ranged from 8.0 cm×4.5 cm to 27.0 cm×16.0 cm. During the surgery, highly selective arterial angiography was used to assist in flap design. The fluorescence development range of the source arteries or perforators of flaps was observed. The blood supply range of the source arteries or perforators of flaps was determined based on the fluorescence development of the skin, and the excision position of the flap was adjusted. The flap incision area ranged from 9.0 cm×6.0 cm to 29.0 cm×16.0 cm. During the surgery, the number of highly selective arterial angiography, the type of source artery of perforators for puncture, and changes in the excision position of flaps were observed and recorded. After surgery, the blood supply and survival of flaps, the healing of wounds and the survival of skin grafts in the flap donor sites, and the angiography-related complications were observed.Results:All the 32 flaps of 29 patients were successfully excised. The highly selective arterial angiography was performed 37 times, including 13 cases of puncture of the oblique branch of the lateral circumflex femoral artery, 6 cases of puncture of the descending branch, 8 cases of double puncture of the oblique and descending branches, and 2 cases of puncture of arteries from other branches. During the surgery, the excision position of 28 flaps did not change, the excision position of 3 flaps moved towards proximal extremity of the thigh, and the excision position of 1 flap moved towards distal extremity of the thigh. All the flaps survived successfully after the surgery, and there was no partial necrosis of the flaps at the proximal or distal ends. The wounds in the flap donor sites healed, and all skin grafts survived. No angiography-related complications occurred.Conclusions:Highly selective arterial angiography can be used to determine the blood supply range of the source artery and perforators of the anterolateral thigh free flaps during the surgery. It can evaluate the blood supply of flaps more intuitively and objectively. Its application in assisting flap design can avoid partial flap necrosis caused by unreasonable preoperative design to a certain extent, and it is safe and reliable.
8.Auxiliary role of Fisher discriminant function based on inflammatory cytokines in diagnosis of depressive disorder
Hailing JIA ; Yongtao YANG ; Lijun CUI ; Yanmei GUO ; Baoping YAN ; Wei CUI ; Xiuli SUN
Sichuan Mental Health 2024;37(4):312-316
Background Currently,the diagnosis of depressive disorder relies on symptomatology,which is greatly influenced by subjective factors such as clinicians' experience.Finding more accurate and reliable quantitative diagnostic methods is currently an urgent problem.Objective To explore the value of Fisher discriminant function based on inflammatory cytokines in the diagnosis of depressive disorder,so as to provide references for clinical diagnosis.Methods A total of 136 patients diagnosed with depressive disorder according to the Diagnostic and Statistical Manual of Mental Disorders,fifth edition(DSM-5)criteria,who underwent inpatient treatment at Hebei Mental Health Center from April 2020 to November 2020,were enrolled as study group.67 healthy participants matched for age and gender,was recruited during the same period.Serum levels of inflammatory cytokine were measured using enzyme-linked immunosorbent assay(ELISA).Fisher discriminant model was employed to establish a discriminant function for inflammatory cytokines exhibiting significant statistical differences between study group and control group,which was then validated.Results The levels of pro-inflammatory cytokines interleukin-1β(IL-1β),interleukin-6(IL-6),interferon-γ(INF-γ)and tumor necrosis factor-α(TNF-α)were higher in the study group compared with control group,with statistically significant differences(U=9.115,5.239,4.431,5.428,P<0.01).Conversely,the levels of anti-inflammatory cytokines interleukin-4(IL-4),interleukin-10(IL-10)and interleukin-13(IL-13)were lower in the study group compared with control group,with statistically significant differences(U=7.398,7.331,7.614,P<0.01).The retrospective test in Fisher discriminant function achieved a correct discrimination rate of 89.66%,and the cross validation achieved a correct discrimination rate of 88.67%.Conclusion The Fisher discriminant function developed in this study may serve as a valuable auxiliary method in the diagnosis of depressive disorder.
9.Analysis of clinical and risk factors of positive conjunctival sac culture before intravitreal injection
Lina SUN ; Yongtao LI ; Meiqi XU ; Guanghua HOU ; Yifeng KE ; Yue ZHENG ; Xina XIN ; Pazo Emmanuel ERIC ; Xinjun REN
Chinese Journal of Ocular Fundus Diseases 2024;40(12):919-923
Objective:To observe and analyze the risk factors of positive conjunctival capsule microbial culture in patients with intravitreal injection treatment (IVT) before treatment.Methods:A prospective study. A total of 1 092 patients who received IVT at the Vitreous Injection Center of Tianjin Medical University Eye Hospital from February 2021 to February 2024 were included in the study. Among them, 539 were males and 553 were females. The age was (62.29±13.61) years. Hypertension and diabetes were 661 and 576 cases, respectively. There were 742 cases of urban residence and 350 cases of rural residence. Three and one days before IVT, 364 patients received antibiotics and 364 patients did not receive antibiotics. Patients' gender, age, history of hypertension and diabetes, pre-IVT antibiotic eye drops use history, and differences in residence (town/country) were collected in detail. Samples were collected after the conjunctival sac was rinsed, and microbial culture was performed. The differences in conjunctival microbial culture positivity rates was compared between those who did not use antibiotic eye drops before IVT, those who used them 1 day before IVT, and those who used them 3 days before IVT. The positive rate of conjunctival sac microbial culture were compared among individuals of different ages, genders, with/without hypertension, with/without diabetes, with different IVT times, and from different living areas (urban/rural). The clinical baseline of positive conjunctival capsule bacterial culture was compared and observed. χ2 test was used to compare the positive rate of conjunctival capsule microbial culture among different clinical baselines. Logistic binary regression analysis was used to analyze the influencing factors. Results:Among the 1 092 patients, 54 cases (4.95%, 54/1 092) were positive for microbial culture of conjunctival sac. There was no significant difference ( P>0.05) in the positive rate of conjunctival sac microbial culture among patients of different ages ( χ2=5.599), gender ( χ2=0.549), residence ( χ2=0.153), with or without hypertension and diabetes ( χ2=3.545, 0.044), and with or without diabetic macular edema ( χ2=0.180). There was no significant difference ( P>0.05) in the positive rate of conjunctival sac microbial culture between patients with different numbers of IVT ( χ2=0.961) or between those who received antibiotic eye drops before IVT and those who did not ( χ2=5.600). Logistic binary regression analysis showed that none of the above factors were risk factors for positive conjunctival capsule microbial culture ( P>0.05). No infective endophthalmitis occurred in all patients during the observation period. Conclusion:The use of antibiotics before IVT is not the decisive factor for positive microbial culture in conjunctival sac.
10.A multicentre cross-sectional study on the correlation of female sexual dysfunction with sex hormones
Rui JU ; Xiangyan RUAN ; Yinmei DAI ; Yu YANG ; Xin XU ; Jiaojiao CHENG ; Yongtao BAI ; Yanhong SHI ; Xingzi YANG ; Yanhua SUN ; Ruiling LI ; Mueck Alfred O.
Chinese Journal of Reproduction and Contraception 2024;44(3):278-284
Objective:To explore the correlation between endogenous sex hormone factors and sexual function in Chinese women.Methods:A total of 711 women who met the criteria were investigated in the Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing Chaoyang District Maternal and Child Health Care Hospital, Beijing Chaoyang District Taiyanggong Community Health Service Centre from April 2020 to March 2021. Sociodemographic data (data split into two groups according to educational background: high school or above and lower high school; and three categories based on marital relationship: satisfied, average, and dissatisfied) and serum levels of sex hormones were collected. Female sexual dysfunction (FSD) was evaluated using the female sexual function index (FSFI). Participants were divided into three groups according to the Stages of Reproductive Aging Workshop (STRAW+10): reproductive stage group, menopausal transition stage group and postmenopausal stage group. Spearman rank correlation analysis and multiple linear regression analyses were performed to determine the correlation between characteristics and scores of FSFI.Results:The multivariate logistic regression analysis revealed that postmenopausal stage, secondary education level, neutral and dissatisfied marital relations were significantly negatively correlated with total FSFI score (all P<0.01). Secondary education level, neutral and dissatisfied marital relations were negatively associated with nearly all FSFI domains apart from pain score (all P<0.05); postmenopausal stage was negatively associated with lubrication, satisfaction and pain (all P<0.05). FSH was negatively associated with all FSFI domain scores (all P<0.05). There was a significantly positive correlation between estradiol and scores of FSFI dimensions other than orgasm and satisfaction (all P<0.05). Total testosterone positively affected the score of total FSFI and desire (all P<0.05); free testosterone was positively related to orgasm ( P<0.05). Conclusion:Estradiol and total testosterone serve as protective factors for female sexual function.

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