BACKGROUND:Exosomes derived from mesenchymal stem cells play pivotal roles in cell communication and epigenetic regulation due to their low immunogenicity and targeted delivery effects,and have been clinically applied in the treatment of various diseases.OBJECTIVE:To review the isolation,purification,identification methods,and application progress of mesenchymal stem cell-derived exosomes,and to facilitate the development of large-scale preparation techniques and clinical translation of mesenchymal stem cell-derived exosomes.METHODS:The Chinese search terms"exosome,mesenchymal stem cells,isolation,purification,characterization,clinical application"and the English search terms"exosome,extracellular vesicles,mesenchymal stem cells,isolation,characterization,application"were used to search the literature published before September 2024 in CNKI,PubMed,and Web of Science databases.Articles with poor relevance to the topic,outdated,or duplicated content were excluded,and finally,109 articles were included for review.RESULTS AND CONCLUSION:(1)This paper reviews recent methods for isolating and purifying exosomes,comparing the characteristics of ultracentrifugation,ultrafiltration,size-exclusion chromatography,polymer precipitation,immunoaffinity,microfluidic methods,and other novel approaches based on their underlying principles.(2)Methods for identifying exosomes can be categorized into physical and biochemical analyses,characterizing exosomes based on their shape,size,and characteristic proteins.(3)Mesenchymal stem cell-derived exosomes have broad applications in multiple fields such as medical aesthetics,wound repair,and cancer treatment,due to their immune-regulatory properties and ability to cross biological barriers.(4)The clinical translation of exosomes faces challenges due to their complex structure,lack of universal isolation techniques,and poor stability,making it difficult to achieve in a short period of time.

BACKGROUND:Exosomes derived from mesenchymal stem cells play pivotal roles in cell communication and epigenetic regulation due to their low immunogenicity and targeted delivery effects,and have been clinically applied in the treatment of various diseases.OBJECTIVE:To review the isolation,purification,identification methods,and application progress of mesenchymal stem cell-derived exosomes,and to facilitate the development of large-scale preparation techniques and clinical translation of mesenchymal stem cell-derived exosomes.METHODS:The Chinese search terms"exosome,mesenchymal stem cells,isolation,purification,characterization,clinical application"and the English search terms"exosome,extracellular vesicles,mesenchymal stem cells,isolation,characterization,application"were used to search the literature published before September 2024 in CNKI,PubMed,and Web of Science databases.Articles with poor relevance to the topic,outdated,or duplicated content were excluded,and finally,109 articles were included for review.RESULTS AND CONCLUSION:(1)This paper reviews recent methods for isolating and purifying exosomes,comparing the characteristics of ultracentrifugation,ultrafiltration,size-exclusion chromatography,polymer precipitation,immunoaffinity,microfluidic methods,and other novel approaches based on their underlying principles.(2)Methods for identifying exosomes can be categorized into physical and biochemical analyses,characterizing exosomes based on their shape,size,and characteristic proteins.(3)Mesenchymal stem cell-derived exosomes have broad applications in multiple fields such as medical aesthetics,wound repair,and cancer treatment,due to their immune-regulatory properties and ability to cross biological barriers.(4)The clinical translation of exosomes faces challenges due to their complex structure,lack of universal isolation techniques,and poor stability,making it difficult to achieve in a short period of time.

AIM:This study aims to investigate the effects of Jiedu-Shengji ointment(JDSJG)on wound healing in diabetic rats by modulating the protein kinase R-like endoplasmic reticulum kinase(PERK)/inositol-requiring enzyme 1(IRE1)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)pathway in the context of endoplasmic reticulum stress(ERS).METHODS:Male Sprague-Dawley rats were randomly assigned to four groups:control group,model group,detoxification muscle ointment group,and sulfadiazine silver cream group.All groups,except the control,were administered an intraperitoneal injection of 45 mg/kg streptozotocin to induce diabetes.The control and model groups received daily applications of normal saline,while the detoxification myogen ointment and sulfadiazine silver cream groups received their respective treatments daily.After dressing changes,wounds were bandaged with sterile gauze.Following 14 d of continuous treatment,wound healing was assessed and healing rates calculated.Histopathologi-cal changes in wound tissues were analyzed using HE staining.Transmission electron microscopy was utilized to observe the number,morphology,and swelling of endoplasmic reticulum in the wound tissues.The expression and distribution of PERK,IRE1 and thioredoxin interacting protein(TXNIP)was assessed by immunohistochemistry,while Western blot was used to measure the levels of apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),TXNIP and NLRP3.ELISA was conducted to quantify the inflammatory cytokines interleukin-18(IL-18)and IL-1β.RE-SULTS:After 14 d of intervention,significant differences were observed in wound tissue parameters across the groups.The model group exhibited a significantly lower wound healing rate compared to the control group(P<0.01),with in-creased wound exudation,poor granulation tissue growth,and elevated the protein levels of PERK,IRE1,TXNIP,CHOP,NLRP3 and ASC(P<0.01),as well as significantly higher levels of IL-1β and IL-18(P<0.01).In contrast,the detoxification muscle ointment group showed a marked improvement in wound healing rate(P<0.01),reduced inflamma-tory exudation,improved granulation tissue growth,and significant decreases in TXNIP expression(P<0.01),along with lower levels of PERK,IRE1,CHOP,ASC and NLRP3(P<0.01).Additionally,the IL-1β and IL-18 were significantly reduced(P<0.01).CONCLUSION:Jiedu Shengji ointment alleviates excessive ERS and mitigates chronic inflammato-ry responses,thereby promoting the healing of diabetic wounds.These effects may be attributed to the inhibition of exces-sive activation of the PERK/IRE1/NLRP3 pathway.

AIM:This study aims to investigate the effects of Jiedu-Shengji ointment(JDSJG)on wound healing in diabetic rats by modulating the protein kinase R-like endoplasmic reticulum kinase(PERK)/inositol-requiring enzyme 1(IRE1)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)pathway in the context of endoplasmic reticulum stress(ERS).METHODS:Male Sprague-Dawley rats were randomly assigned to four groups:control group,model group,detoxification muscle ointment group,and sulfadiazine silver cream group.All groups,except the control,were administered an intraperitoneal injection of 45 mg/kg streptozotocin to induce diabetes.The control and model groups received daily applications of normal saline,while the detoxification myogen ointment and sulfadiazine silver cream groups received their respective treatments daily.After dressing changes,wounds were bandaged with sterile gauze.Following 14 d of continuous treatment,wound healing was assessed and healing rates calculated.Histopathologi-cal changes in wound tissues were analyzed using HE staining.Transmission electron microscopy was utilized to observe the number,morphology,and swelling of endoplasmic reticulum in the wound tissues.The expression and distribution of PERK,IRE1 and thioredoxin interacting protein(TXNIP)was assessed by immunohistochemistry,while Western blot was used to measure the levels of apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),TXNIP and NLRP3.ELISA was conducted to quantify the inflammatory cytokines interleukin-18(IL-18)and IL-1β.RE-SULTS:After 14 d of intervention,significant differences were observed in wound tissue parameters across the groups.The model group exhibited a significantly lower wound healing rate compared to the control group(P<0.01),with in-creased wound exudation,poor granulation tissue growth,and elevated the protein levels of PERK,IRE1,TXNIP,CHOP,NLRP3 and ASC(P<0.01),as well as significantly higher levels of IL-1β and IL-18(P<0.01).In contrast,the detoxification muscle ointment group showed a marked improvement in wound healing rate(P<0.01),reduced inflamma-tory exudation,improved granulation tissue growth,and significant decreases in TXNIP expression(P<0.01),along with lower levels of PERK,IRE1,CHOP,ASC and NLRP3(P<0.01).Additionally,the IL-1β and IL-18 were significantly reduced(P<0.01).CONCLUSION:Jiedu Shengji ointment alleviates excessive ERS and mitigates chronic inflammato-ry responses,thereby promoting the healing of diabetic wounds.These effects may be attributed to the inhibition of exces-sive activation of the PERK/IRE1/NLRP3 pathway.

OBJECTIVE To explore the effect of Zi Zhu Ointment combined with herbal fumigation on wound heal-ing,oxidative stress and microribonucleic acid(miR)-200b of the patients with diabetic foot infection.METHODS A total of 102 patients with diabetic foot infection who were treated in Heping Hospital Affiliated to Changzhi Medical College from Sep.2020 to Sep.2023 were enrolled in the study and were divided into the control group with 48 cases and the study group with 54 cases according to the treatment program.The control group was trea-ted with conventional therapy and herbal fumigation,the study group was given additional Zi Zhu Ointment on ba-sis of the treatment of the control group,and both groups were persistently treated for 1 month.The clinical cura-tive effect,wound healing status,hemodynamics indexes of dorsal foot,pain score,oxidative stress indexes and miR-200b expression levels were observed and compared between the two groups of patients.RESULTS The effec-tive rate of clinical treatment of the study group was higher than that of the control group after the treatment(Z=2.138,P=0.033).The wound area of the study group was(0.42±0.05)cm2 after the treatment,smaller than that of the control group;the cover area of neoplastic granulation tissue of the study group was(16.87±3.21)cm2,larger than that of the control group;the coverage fraction of the neoplastic granulation tissue of the study group was(55.23±6.17)％,greater than that of the control group;there were significant differences between the two groups(P＜0.05).The blood vessel diameter of dorsal foot of the study group was greater than that of the control group after the treatment,the blood flow velocity of the study group was greater than that of the control group,and the visual analogue scale(VAS)score of the study group was lower than that of the control group(P＜0.05).There were no significant differences in the expression levels of advanced oxidative protein product(AOPP),superoxide dismutase(SOD),malondialdehyde(MDA)and miR-200b between the two groups of pa-tients before the treatment;the expression levels of AOPP,MDA and miR-200b of the study group were lower than those of the control group after the treatment,while the SOD level of the study group was higher than that of the control group(P＜0.05).CONCLUSIONS Zi Zhu Ointment combined with herbal fumigation can remarkably improve the clinical curative effect on the patients with diabetic foot infection,promote the wound healing,im-prove the arterial hemodynamics of dorsal foot,and relieve the pain.The mechanism may be associated with the improvement of oxidative stress response and downregulated expression level of miR-200b.

OBJECTIVE To explore the effect of Zi Zhu Ointment combined with herbal fumigation on wound heal-ing,oxidative stress and microribonucleic acid(miR)-200b of the patients with diabetic foot infection.METHODS A total of 102 patients with diabetic foot infection who were treated in Heping Hospital Affiliated to Changzhi Medical College from Sep.2020 to Sep.2023 were enrolled in the study and were divided into the control group with 48 cases and the study group with 54 cases according to the treatment program.The control group was trea-ted with conventional therapy and herbal fumigation,the study group was given additional Zi Zhu Ointment on ba-sis of the treatment of the control group,and both groups were persistently treated for 1 month.The clinical cura-tive effect,wound healing status,hemodynamics indexes of dorsal foot,pain score,oxidative stress indexes and miR-200b expression levels were observed and compared between the two groups of patients.RESULTS The effec-tive rate of clinical treatment of the study group was higher than that of the control group after the treatment(Z=2.138,P=0.033).The wound area of the study group was(0.42±0.05)cm2 after the treatment,smaller than that of the control group;the cover area of neoplastic granulation tissue of the study group was(16.87±3.21)cm2,larger than that of the control group;the coverage fraction of the neoplastic granulation tissue of the study group was(55.23±6.17)％,greater than that of the control group;there were significant differences between the two groups(P＜0.05).The blood vessel diameter of dorsal foot of the study group was greater than that of the control group after the treatment,the blood flow velocity of the study group was greater than that of the control group,and the visual analogue scale(VAS)score of the study group was lower than that of the control group(P＜0.05).There were no significant differences in the expression levels of advanced oxidative protein product(AOPP),superoxide dismutase(SOD),malondialdehyde(MDA)and miR-200b between the two groups of pa-tients before the treatment;the expression levels of AOPP,MDA and miR-200b of the study group were lower than those of the control group after the treatment,while the SOD level of the study group was higher than that of the control group(P＜0.05).CONCLUSIONS Zi Zhu Ointment combined with herbal fumigation can remarkably improve the clinical curative effect on the patients with diabetic foot infection,promote the wound healing,im-prove the arterial hemodynamics of dorsal foot,and relieve the pain.The mechanism may be associated with the improvement of oxidative stress response and downregulated expression level of miR-200b.

Breast cancer has become the malignant tumor with the highest incidence rate among women, of which human epidermal growth factor receptor 2 (HER2) positive breast cancer accounts for about 15%-20%. With the development of anti HER2 targeted drugs, the survival and prognosis of HER2 positive breast cancer has significantly benefited. About 45%-55% of breast cancer patients have low HER2 expression, and these patients usually do not receive anti HER2 treatment. However, there is a significant difference between the biological behavior and prognosis of breast cancer with low HER2 expression and breast cancer with zero HER2 expression. It is helpful to differentiate and adopt corresponding treatment strategies to improve the prognosis of patients. At present, there have been many advances in targeted therapy of breast cancer with low HER2 expression, which provides a useful reference for precision treatment of breast cancer with low HER2 expression.

Erythromycin is a macrolide antibiotic produced by Saccharopolyspora erythraea. Its yield is greatly affected by the fermentation conditions and the bioreactor configurations. In this study, a novel scale-up method for erythromycin fermentation was developed based on computational fluid dynamics (CFD) and time constant analysis. Firstly, the dissolved oxygen (DO) was determined as a key parameter according to the physiological properties of S. erythraea cultivated in a 50 L bioreactor. It was found that the time constant of oxygen supply (t_mt) in a 500 m³ bioreactor should be less than 6.25 s in order to satisfy the organism's oxygen uptake rate (OUR). Subsequently, a 500 m³ bioreactor was designed using the time constant method combined with empirical correlations. The impeller combination with one BDT8 impeller at bottom and two MSX4 impellers at upper part was determined, and then validated by numerical simulation. The results indicated that the t_mt of the bioreactor (< 6.25 s) and the fluid properties, including gas hold-up, shear stress and fluid vector, met the requirements of erythromycin fermentation. Finally, the industrial production of erythromycin in the 500 m³ showed the design method was applicable in large scale fermentation.
Erythromycin ; Saccharopolyspora/genetics* ; Bioreactors ; Fermentation ; Anti-Bacterial Agents

Bioreactors have been central in monoclonal antibodies and vaccines manufacturing by mammalian cells in suspension culture. Numerical simulation of five impeller combinations in a stirred bioreactor was conducted, and characteristics of velocity vectors, distributions of gas hold-up, distributions of shear rate in the bioreactor using 5 impeller combinations were numerically elucidated. In addition, genetically engineered CHO cells were cultivated in bioreactor installed with 5 different impeller combinations in fed-batch culture mode. The cell growth and antibody level were directly related to the maximum shear rate in the bioreactor, and the highest viable cell density and the peak antibody level were achieved in FBMI3 impeller combination, indicating that CHO cells are sensitive to shear force produced by impeller movement when cells were cultivated in bioreactor at large scale, and the maximum shear rate would play key roles in scaling-up of bioreactor at industrial scale.
Animals ; Batch Cell Culture Techniques ; Bioreactors ; standards ; CHO Cells ; Cell Count ; Computer Simulation ; Cricetinae ; Cricetulus ; Industrial Microbiology ; instrumentation ; methods

Objective To observe the effects of electro-acupuncture on loading of lower limb joints in patients with knee osteoarthritis (KOA) during stair climbing and explore the related biomechanical mechanism. Methods Forty patients with KOA were randomly assigned, with 20 patients in observation group (electro-acupuncture group, EA group) and 20 patients in control group (superficial acupuncture group, SA group). Finally 18 patients in each group completed the study. In observation group, seven knee acupuncture points were chosen and patients were connected with electro-acupuncture instrument; while in control group, the electro-acupuncture instrument was connected but not electrified after superficial acupuncture at non-acupoint points. The three-dimensional gait analysis system was used to assess the biomechanical characteristics during stair climbing before and after treatment, including peak vertical force (PFz), vertical impulse (IFz) and symmetry index (SI%). Results After 3 weeks of treatment in EA group, PFz of the right foot during stair ascent and PFz of the left foot during stair descent increased (P＜0.05); IFz of both feet during stair ascent and IFz of the right foot during stairs descent significantly decreased (P＜0.05); no significant differences were found in SI% of peak and impulse (P＞0.05). In SA group, only SI of impulse during stairs ascent increased (P＜0.05). There was no significant difference between two groups before and after treatment (P＞0.05). Conclusions Electro-acupuncture can effectively improve the joint load capacity and reduce the dynamic cumulative load of patients w