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.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.
3.Evaluation of conjunctival sac culture findings prior to intravitreal injection
Xinjun REN ; Yifeng KE ; Liangzhang TAN ; Eric Emmanuel PAZO ; Yongtao LI ; Xiaorong LI
Chinese Journal of Ocular Fundus Diseases 2024;40(9):687-692
Objective:To observe the differences in the positive rate of conjunctival sac microbial culture after different methods of preventing infection before intravitreal injection (IVI).Methods:A prospective case-control study. A total of 1 200 participants with fundus diseases who received IVI injection at Tianjin Medical University Eye Hospital from July 2021 to December 2023 were included. Patients were randomly divided into 6 groups according to eye spot with antibiotic solution 3, 1 and 0 days before IVI and local eye disinfection with povidone-iodine (PVI) 3 min and 30 s before IVI: the first 3 days of antibiotics+3 min PVI group, the first 1 day of antibiotics+3 min PVI group, the first 0 days of antibiotics+3 min PVI group, the first 3 days of antibiotics+30 s PVI group, the first 1 day of antibiotics+30 s PVI group, the first 0 days of antibiotics+30 s PVI group, there were 200 cases in each group. Microbial sampling and cultivation of conjunctival sac were conducted before IVI to compare the differences in positive rates among different groups. Multiple group comparisons were conducted using one-way analysis of variance. The comparison of count data is conducted using χ2 test. Results:Among the 1 200 patients, there were 566 males and 634 females. Age (62.59±13.44) years old. There were 397 cases of diabetes and 482 cases of hypertension. IVI frequency (2.35±2.34). 64 cases were positive for conjunctival sac culture before IVI. The age ( F=1.468), sex composition ratio ( χ2=2.876), diabetes ( χ2=10.002), hypertension ( χ2=6.019), times of IVI ( χ2=4.507), and positive rate of conjunctival sac bacterial culture ( χ2 =6.272) of patients in each group had no statistical significance ( P>0.05). Using the duration of antibiotic application before IVI as a stratified factor, there was no statistically significant difference in the positive rate of conjunctival sac culture between groups with different durations of antibiotic application before IVI [ χ2=0.414, P=0.52, combined odds ratio ( OR)=0.819, 95% confidence interval ( CI) 0.493-1.360]. Using the duration of PVI application as a stratified factor, there was no statistically significant difference in the positive rate of conjunctival sac culture between different PVI disinfection times [ χ2=0.000, P=1.000, combined OR=1.00, 95% CI 0.503-1.988]. Conclusions:Pre IVI treatment with 0.5% PVI for 30 s can inhibit the growth of microbial colonies in the conjunctival sac. The application of local antibiotic eye fluid in the anterior eye of IVI cannot reduce the positive rate of conjunctival sac bacteria.
4.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.
5.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.
6.Effects of intravenous injection of human umbilical cord mesenchymal stem cells-derived small extracellular vesicles on experimental autoimmune uveitis in mice
Yongtao LI ; Yanan DUAN ; Huan LI ; Zhihui ZHANG ; Xinjun REN ; Xiaorong LI ; Xiaomin ZHANG
Chinese Journal of Experimental Ophthalmology 2021;39(11):949-956
Objective:To explore the therapeutic effects of intravenous injection of small extracellular vesicles (sEVs) derived from human umbilical cord mesenchymal stem cells (MSCs) on experimental autoimmune uveitis (EAU) in mice.Methods:MSCs from human umbilical cord were cultured and the supernatant was collected.The sEVs were isolated by ultracentrifugation method and a NanoSight instrument was used to analyze the particle size.The expression of surface markers sEVs, CD9, CD81 and CD63 was determined via Western blot.The morphology of sEVs was observed with a transmission electron microscope.Forty-eight 7-week-old female C57BL/6 mice were seclected to establish the EAU model through immunization with interphotoreceptor retinoid-binding protein peptide 651-670 (IRBP 651-670). The mice were divided into sEVs treatment group and phosphate buffer solution (PBS) control group using a random number table, with 24 mice in each group.The mice in the sEVs treatment group were injected with 50 μg of MSCs-derived sEVs via tail vein on the 11th day after modeling.In the PBS control group, the mice were injected with the same volume of PBS.Six mice in each group were randomly selected to observe the inflammation of the retina after mydriasis with an ophthalmoscope every other day from 8th day following modeling and the inflammation scores were evaluated.Six mice were randomly selected and sacrificed on the 14th day and 6 on the 18th day following modeling in each group, and both eyeballs of the mice were enucleated.Retinal tissue sections of the 6 mice sacrificed on the 18th day were stained with hematoxylin-eosin and the pathological scores were evaluated.The infiltration of helper T 1 (Th1) cells and Th17 cells in the eyeballs of the 6 mice sacrificed on the 18th day following modeling was detected by flow cytometry.T cells were isolated from spleen and lymph nodes of the 6 mice sacrificed on the 14th day, and the proliferation of T cells under different concentrations of IRBP 651-670 (0, 1, 10 and 20 μg/ml) was detected using a 5-bromodeoxyuridine (BrdU) method.To further study the effects of MSCs-derived sEVs on Th1/Th17 cells differentiation, naive T cells of spleen from another 3 normal mice were isolated by magnetic bead negative sorting and incubated with 10 μg/ml MSCs-derived sEVs or 10 μg/ml PBS, and then were cultured under Th1/Th17 cell differentiation conditions, respectively.Flow cytometry was used to measure the differentiation of naive T cells into Th1/Th17 cells.This study protocol complied with the regulations of the care and use of laboratory animals in China and was approved by an Ethics Committee of Tianjin Medical University (No.TJYY2019103022). Results:The isolated human MSCs-derived sEVs was with an average diameter of (102.4±33.6) nm and showed a double-layer membrane vesicle structure under the transmission electron microscope.The CD9, CD63 and CD81 proteins were highly expressed in sEVs.The inflammation scores of the sEVs treatment group were significantly lower than those in the control group on day 14, 16, 18, 20 and 22 after modeling (all at P<0.05). The pathological score of mice in the sEVs treatment group was significantly lower than that of PBS control group on the 18th day following modeling ( P<0.05). The flow cytometry results showed that on day 18 after modeling, the proportions of Th1 and Th17 cells in eyeballs in the sEVs treatment group were (15.55±2.03)% and (15.67±2.15)%, respectively, which were significantly lower than (21.35±0.72)% and (20.90±1.10)% in the PBS control group ( t=6.58, 5.31; both at P<0.01). BrdU results showed that when the IRBP 651-670 concentration was 20 μg/ml, the T cell proliferation ability in the sEVs treatment group was inhibited obviously compared with the control group ( P<0.05). The proportions of naive T cells differentiated into Th1 cells and Th17 cells in the sEVs treatment group were (28.15±1.32)% and (11.60±2.23)% respectively, which were significantly lower than (31.58±1.75)% and (23.52±1.76)% of the PBS control group, and the differences were statistically significant ( t=3.93, 10.26; both at P<0.05). Conclusions:Intravenous injection of human umbilical cord MSCs-derived sEVs can reduce the inflammation in EAU mice.The mechanism may be related to inhibiting the differentiation of naive T cells to Th1 and Th17 cells, and reducing the infiltration of Th1 and Th17 cells in the eyeballs.
7.Rate of capsular contracture among different breast implants in the last two decades: a meta-analysis
Peipei WANG ; Ziyang FU ; Yongtao LI ; Jizhen REN ; Yanjin WANG ; Shouju LI
Chinese Journal of Medical Aesthetics and Cosmetology 2021;27(6):513-516
Objective:To study the rate of capsule contracture after operation of the textured surfaces breast implants and the smooth surfaces breast implants, to provide evidence for plastic surgeons to select the type of breast implants during breast augmentation.Methods:This study started from January 2018 to May 2019. Chinese and English databases including Wanfang Science and Technology Periodical Full-text Database, VIP Chinese Science and Technology Periodical Full-text Database (VIP) and CNKI, PubMed, Cochrane Library, Web of Science, Science Drirect Online were searched by computer. Some relevant studies were collected for this meta-analysis.Results:We identified 9 studies including a total of 13165 subjects for the meta-analysis. The OR value of the study was 0.43 (95% CI: 0.35, 0.51), and the incidence rate of capsule contracture in the experimental group was lower than that in the control group. In cumulative meta-analysis and sensitivity test, the conclusion was stable. And there was no publication bias found by Egger regression test. Conclusions:The textured surfaces breast implants are better than the smooth surfaces breast implants in terms of incidence rate of capsule contracture after augmentation mammoplasty.
8. HBsAg loss with Pegylated-interferon alfa-2a in hepatitis B patients with partial response to nucleos(t)-ide analog: new switch study
Peng HU ; Jia SHANG ; Wenhong ZHANG ; Guozhong GONG ; Yongguo LI ; Xinyue CHEN ; Jianning JIANG ; Qing XIE ; Xiaoguang DOU ; Yongtao SUN ; Yufang LI ; Yingxia LIU ; Guozhen LIU ; Dewen MA ; Xiaoling CHI ; Hong TANG ; Xiaoou LI ; Yao XIE ; Xiaoping CHEN ; Jiaji JIANG ; Ping ZHA ; Jinlin HOU ; Zhiliang GAO ; Huimin FAN ; Jiguang DING ; Dazhi ZHANG ; Hong REN
Chinese Journal of Hepatology 2018;26(10):756-764
Objective:
Hepatitis B surface antigen (HBsAg) loss is seldom achieved with nucleos(t)ide analog (NA) therapy in chronic hepatitis B patients but may be enhanced by switching to finite pegylated-interferon (Peg-IFN) alfa-2a. We assessed HBsAg loss with 48- and 96-week Peg-IFN alfa-2a in chronic hepatitis B patients with partial response to a previous NA.
Methods:
Hepatitis B e antigen (HBeAg)-positive patients who achieved HBeAg loss and hepatitis B virus DNA < 200 IU/mL with previous adefovir, lamivudine or entecavir treatment were randomized 1:1 to receive Peg-IFN alfa-2a for 48 (
9.Application of Position Number in the Drug Management in Pharmacy Department
Chunling REN ; Yi LI ; Jinpeng ZHANG ; Zhanfang CHEN ; Xiaomei MO ; Yongtao LENG
China Pharmacy 2015;(19):2686-2688
OBJECTIVE:To provide reference for the application of position number in the pharmacy drug management. METHODS:Three-dimensional coding method was used for coding the position number. The mentioned method was combined with hospital information management system (HIS) for the out of storage,deployment and inventory. Memory field assumptions method was used to compare the size of field memorized by pharmacist in inpatient pharmacy before and after management of posi-tion number. Sampling controlled trial was conducted to compare the drugs deployment time and walking distance of pharmacists in inpatient pharmacy and drug storehouse before and after coding management of position number. RESULTS:After management of coding management in inpatient pharmacy,the memory required field was decreased from 1 028 to 25,deployment time of pharma-cists was decreased from(36.57±0.82)min to(24.20±0.33)min,and the walking distance was decreased from(79.17±0.29)m to(38.59±0.56)m. After management of coding management in drug storehouse,deployment time of pharmacists was decreased from(61.86±0.44)min to(47.18±0.63)min,and the walking distance was decreased from(129.53±0.58)m to(68.97±0.32) m. CONCLUSIONS:The drug coding management of position number can improve the deployment efficiency and reduce the brain and physical quantity of pharmacists.
10.External use of recombinant human granulocyte-macrophage colony stimulating factor hydrogel to repair thick skin graft donor sites
Chao LI ; Shouju LI ; Yongtao LI ; Ziyang FU ; Changyin REN
Chinese Journal of Tissue Engineering Research 2015;(34):5458-5462
BACKGROUND:There are less reports about the external use of recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) hydrogel to repair thick skin graft donor sites. By now, relevant self-control studies have not been retrieved. OBJECTIVE:To observe the effect of rhGM-CSF on the repair of thick skin graft donor sites. METHODS:Sixty patients with burns and scar hyperplasia undergoing autologous thick skin grafting were enroled, 47 males and 13 females, aged 18-65 years. The thigh was selected as donor sites. According to the depth of donor sites, the patients were divided into 0.4 mm and 0.55 mm groups, with 30 cases in each group. Wounds on the symmetric areas with equal area and same depth were selected or wounds with same depth were selected and divided equaly. The wounds were randomly assigned into treatment group and control group. The treatment group was treated with rhGM-CSF hydrogel externaly; the control group was only given vaseline dressing. At postoperative 3, 7, 10, 14 days, the fresh dressing was changed. Then, the wound appearance, healing time, healing rate and adverse effects were observed in the two groups. RESULTS AND CONCLUSION:At 14 days after operation, the wound surface was smoother and the pigmentation was relatively less in the treatment group compared with the control group; the degree of wound pain
was less in the treatment group than the control group during dressing change (P < 0.05). At 10 and 14 days after operation, the healing rate and healing time were better in the treatment group than the control group (P < 0.05). No general malaise or hypersensitivity cases were reported, and local issue hyperplasia was also not found. Al the above indicate that the external use of the rhGM-CSF hydrogel can evidently shorten the healing time and improve the healing condition when it is applied in the thick skin graft donor sites.

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