BACKGROUND: A general standard has not been established for the extraction and purification of adipose-derived stem cells (ADSCs). An erythrocyte lysis step for stromal vascular fraction is the commonly used method in the procedure for ADSCs isolation. However, there is a lack of evidence on whether this step will have adverse effects on human ADSCs (hADSCs). OBJECTIVE: To test the efficiency of two hADSCs isolation methods, which are erythrocyte-lysis method based on ammonium chloride and non-lysis method. Moreover, the biological characteristics of the hADSCs isolated by the two methods were also compared. METHODS: After collagenase digestion of lipoaspirate, erythrocyte lysis buffer was used to purify stromal vascular fraction in erythrocyte-lysis method, while in non-lysis method the buffer was not used. A Muse™ cell analyzer was used to assess living cell counting and proportion of stromal vascular fraction in both methods. Then hADSCs were cultured to the second passage for next testing. Cell morphology was observed under light microscope. Cell phenotype was detected by flow cytometry. Cell counting kit-8 was used to evaluate cell proliferation. Oil red O staining and alizarin staining were used to evaluate adipogenic and osteogenic ability of hADSCs after adipogenic and osteogenic induction. This study was approved by the Ethics Committee of the Plastic Surgery Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, and informed consents were signed by all participants. RESULTS AND CONCLUSION: (1) Compared with the erythrocyte lysis group, hADSCs obtained in the non-lysis group contained a larger number and a larger percentage of non-erythrocyte living cells. (2) The two groups of hADSCs were spindle-shaped and arranged as a fish shape. (3) The cell phenotypes of both groups met the phenotypic requirements for human mesenchymal stem cells. (4) The cell proliferation in the non-lysis group was faster than that in the erythrocyte lysis group, while there was no difference in the adipogenic and osteogenic abilities between the two groups. In conclusion, the use of erythrocyte lysis buffer reduces the isolation efficiency of hADSCs and inhibits cell proliferation. The non-lysis isolation method does not affect phenotypes, adipogenic and osteogenic ability of hADSCs. Therefore, it is not recommended to implement erythrocyte lysis during the isolation of hADSCs.

In recent years, breast implant-associated anaplastic large cell lymphoma (BIA-ALCL) has become the research focus of many specialists in the field. This study introduces the epidemiological features of BIA-ALCL, reviewing the characteristics of prosthesis implantation and prosthesis and briefly describing its clinical manifestations, diagnosis, and treatment. Additionally, the research progress of its disease mechanism was summarized. Altogether, BIA-ALCL not only requires plastic surgeons to be vigilant and to regulate the collection and reporting of cases, but also requires them to use a multidisciplinary approach for conducting thorough research.

Despite decades of research,cancer continues to be a major global health concern.The human mouth appears to be a multiplicity of local environments communicating with other organs and causing diseases via microbes.Nowadays,the role of oral microbes in the development and progression of cancer has received increasing scrutiny.At the same time,bioengineering technology and nanotechnology is growing rapidly,in which the physiological activities of natural bacteria are modified to improve the therapeutic efficiency of cancers.These engineered bacteria were transformed to achieve directed genetic reprogramming,selective functional reorganization and precise control.In contrast to endotoxins produced by typical genetically modified bacteria,oral flora exhibits favorable biosafety characteristics.To outline the current cognitions upon oral microbes,engineered microbes and human cancers,related literatures were searched and reviewed based on the PubMed database.We focused on a number of oral microbes and related mechanisms associated with the tumor microenvironment,which involve in cancer occurrence and development.Whether engineering oral bacteria can be a possible application of cancer therapy is worth consideration.A deeper understanding of the relationship between engineered oral bacteria and cancer therapy may enhance our knowledge of tumor pathogenesis thus providing new insights and strategies for cancer prevention and treatment.

With the rapid development of biotechnology, therapeutic peptide has been a hot area in the central nervous system drugs due to its features of easy to design and target specificity. However, therapeutic peptide is difficult to cross the blood brain barrier into the central nervous system and target cells, coupled with its in vivo instability, which seriously restricts its application in central nervous system diseases. This review focuses on the progress of therapeutic peptides across the blood brain barrier targeting the central nervous system, compares and analyses the methods of increasing therapeutic peptides penetration, specificity and stability in combination with other molecules, in order to provide help for the development of central nervous system drugs.

In recent years,3D bioprinting technology has developed rapidly,becoming an essential tool in the fields of cancer research,tissue engineering,disease modeling and mechanistic studies.This paper reviewed the fundamental principles of bioprinting technology and its current applications in cancer research and tissue engineering.Bioprinting is an additive manufacturing technology that constructs complex three-dimensional tissue structures by digitally controlling the layer-by-layer deposition of biomaterials and living cells.The core steps of bioprinting include designing a 3D model,selecting appropriate bioprinting techniques and materials,printing layer by layer,followed by post-processing involving cell culture and functionalization.In cancer research,3D bioprinting can create complex tumor models that simulate the tumor microenvironment,revealing new mechanisms of tumor initiation and progression.Traditional in vitro models,such as 2D cell cultures or animal models,often fail to accurately replicate the complexity of human tumors.However,3D bioprinted tumor models,which mimic the dynamic interactions between tumor cells and their environment such as immune cells,stroma and blood vessels,offer a more biomimetic platform for studying tumor growth,invasion and metastasis.These models provide a research platform that closely mirrors actual tumor behavior.Additionally,Bioprinted models and scaffolds can be leveraged in personalized precision therapies by efficiently constructing patient-specific 3D models from their own cells.These models enable the prediction of patient's sensitivity to drugs and radiotherapy.Additionally,localized scaffolds can be developed to meet individual patient needs,allowing for the formulation of appropriate drug types and dosages.Furthermore,3D-printed scaffolds can support drug delivery by targeting specific areas,reducing drug-related side effects.They can also be used to facilitate local immunotherapy,cytokine therapy,cancer vaccines,and chimeric antigen receptor cell therapy,enhancing therapeutic outcomes.In tissue engineering,traditional tissue repair methods often struggle to address the complex requirements of constructing intricate tissue structures.3D bioprinting offers a novel solution by enabling the creation of complex tissue architectures and promoting tissue regeneration.Basic tissues,such as bone,cartilage and skin,which have higher regenerative capacities,are gradually being incorporated into clinical practice.Significant progress has also been made in the repair and reconstruction of more complex organs like the liver and heart,though considerable challenges remain before these advancements can be fully translated into clinical applications.Finally,this paper discussed the current challenges and future directions of 3D bioprinting in these fields,aiming to provide reference for researchers.

BACKGROUND: Oral submucous fibrosis (OSF) is a chronic disease with carcinogenic tendency that poses a non-negligible threat to human health. Exosomes derived from human adipose mesenchymal stem cells (ADSC-Exo) reduces visceral and cutaneous fibroses, but their role in OSF has received little attention. The aim of this study was to investigate the effects of ADSC-Exo on OSF and elucidate the mechanism. METHODS: In brief, ADSCs were extracted from adipose tissues and subjected to flow cytometry and induction culture. Fibroblasts were isolated from human buccal mucosa and subjected to immunofluorescence. Myofibroblasts were obtained from fibroblasts induced by arecoline and identified. Immunofluorescence assay confirmed that myofibroblasts could take up ADSC-Exo. The effects of ADSC-Exo on the proliferative and migratory capacities of myofibroblasts were examined using the Cell Counting Kit-8 and scratch assay. Real-time quantitative polymerase chain reaction (qPCR) was performed to evaluate mothers against decapentaplegic homolog 2 (Smad2), Smad3, Smad7, collagen type 1 (Col1), Col3, alpha smooth muscle actin (α-SMA), fibronectin, and vimentin. Western blotting was performed to detect phospho (p)-Smad2, Smad2, p-Smad2/3, Smad2/3, Smad7, Col1, Col3, α-SMA, fibronectin, and vimentin. Furthermore, the dual-luciferase reporter assay was performed to prove that miR-181a-5p in ADSC-Exo directly inhibited the expression of Smad2 mRNA to regulate the transforming growth factor beta (TGF-β) pathway. We also performed qPCR and western blotting to verify the results. RESULTS: ADSC-Exo could promote the proliferation and migration of myofibroblasts, reduce the expressions of p-smad2, Smad2, p-smad2/3, Smad2/3, Col1, αSMA, fibronectin, and vimentin and elevated the levels of Smad7 and Col3. In addition, miR-181a-5p was highly expressed in ADSC-Exo and bound to the 3'-untranslated region of Smad2. ADSC-Exo enriched with miR-181a-5p reduced collagen production in myofibroblasts and modulated the TGF-β pathway. CONCLUSIONS ADSC-Exo promoted the proliferative and migratory capacities of myofibroblasts and inhibited collagen deposition and trans-differentiation of myofibroblasts in vitro. miR-181a-5p in exosomes targets Smad2 to regulate the TGF-β pathway in myofibroblasts. ADSC-Exo perform antifibrotic actions through the miR-181a-5p/Smad2 axis and may be a promising clinical treatment for OSF.

Objective To explore the regularity of traditional Chinese medicine (TCM) in the treatment of postembolization syndrome (PES) after transcatheter arterial chemoembolization (TACE). Methods CNKI, WANFANG and VIP were used as data sources to search the journals and literatures related to TCM treatment from January 2000 to December 2021. Then, relevant TCM formula or Chinese patent medicines preparations were screened out. The Chinese medicinal materials contained were entered into Excel 2019 table database, and the data were analyzed by SPSS Statistics 21.0 and SPSS Modeler 18.0 statistical software. Results 86 qualified prescriptions were included, containing 181 Chinese medicinal materials, with a total frequency of 942 times. Of the 181 Chinese herbs included, there were 28 herbs with frequency ≥10%, with a total frequency of 587. The top 5 Chinese medicinal materials of frequency were licorice, Poria, Atractylodes, Bupleurum and Astragalus. Among the efficacy classifications, tonifying deficiency drugs, heat-clearing drugs and diuretics were most used. In four properties and five tastes, the top three of four properties were warm, flat and cold, and the top three of five tastes were sweet, bitter and pungent. In the classification of meridians, the first three meridians were spleen meridian, lung meridian and liver meridian. 30 association rules were obtained in association rules analysis, 11 common factors were obtained by factor analysis, 6 clustering combinations were obtained by cluster analysis, and 4 commonly used drug combinations were obtained. Conclusion The prescription drugs for the treatment of PES after TACE were mainly tonic drugs, heat-clearing drugs and diuresis and dampness-draining drugs. The treatment methods were mainly invigorating spleen and replenishing qi, clearing heat and dampness and detoxification.

Objective:To explore the effects of general anesthesia and combined spinal and epidural anesthesia on inflammatory factors and pain in patients with osteoarthritis after total knee arthroplasty.Methods:A total of 84 patients with osteoarthritis who underwent unilateral total knee arthroplasty in Hulunbuir People's Hospital from January 2020 to May 2021 were selected as the research subjects. They were randomly divided into general anesthesia group (40 cases) and combined spinal and epidural anesthesia group (44 cases). Venous blood samples of 5 ml were collected before operation and 6, 24, 48 hours after operation, and the contents of inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6] in serum were determined by enzyme-linked immunosorbent assay (ELISA). The visual analogue pain scale (VAS) of the two groups at 30 min, 6, 24 and 48 hours after operation was compared.Results:At 6 and 24 hours after operation, TNF-α, IL-1β and IL-6 levels in the combined spinal and epidural anesthesia group were lower than those in the general anesthesia group ( t = 4.17, 3.85, 8.95, 10.98, 10.04, 9.87, P < 0.05). There were significant differences in the levels of TNF-α, IL-1β and IL-6 at different time points between the general anesthesia group and combined spinal and epidural anesthesia group ( F = 271.67, 149.26, 81.70, 189.36, 102.44, 157.32, P < 0.001). At 6 and 24 hours after operation, the VAS scores of patients in the combined spinal and epidural anesthesia group were significantly lower than those in the general anesthesia group ( t = 6.60, 3.66, P < 0.05). There were statistically significant differences in VSA scores between the two groups at different time points ( F = 67.47, 52.37, P < 0.05). Conclusion:The effect of combined spinal and epidural anesthesia is significantly higher than general anesthesia in inhibiting the expression of TNF-α, IL-1β and IL-6 in patients with osteoarthritis after operation, and the effect of analgesia is obvious.

Objective:The aim of this study is to compare the effects of age on the biological properties of adipose-derived stem cells(ASCs) and fat survival of ASC-assisted lipotransfer. To identify the effect of age factors on the biological characteristics of human ASCs and compare the effects of ASCs-assisted subcutaneous fat transplantation/lipotransfer on nude mice at different ages.Methods:Human lipoaspirates were obtained from 30 healthy female patients (aged from 18 to 65 years) acquiring the abdominal liposuction. Samples were divided into three groups according to donor age: group A, 18-29 years; group B, 30-49 years; group C, 50-65 years. Stromal vascular fraction cells were isolated from the harvested adipose tissue using collagenase. The yield and cell viability of SVF were tested using the Muse cell count and viability assay. ASCs were cultured and harvested at the second passage. MSC surface markers of ASCs were examined by the flow cytometry. The cell proliferation of ASCs from different age was determined by the CCK-8 assay. The scratch test was used for assessing the ASCs migration ability. The adipogenic differentiation potential of ASCs was analyzed by induction of lipid formation in vitro. The expression levels of PPAR-γ and CEBP-α genes were detected by RT-PCR assay. The survival of adipocytes in the grafts was analyzed by perilipin-A immunofluorescence staining. The fat survival of ASCs-enriched grafts from different age was measured in animal models. The weight and residual volume of fat grafts were compared in different groups after three months. The histologic analysis was evaluated by cell integrity and necrosis tissue in fat grafts. The vessel density was measured using the CD31 immunohistochemical staining. Data were analyzed by SPSS software version 21.0 with one-way ANOVA to compare the difference of multiple groups. A value of P< 0.05 was considered statistically significant. Results:The yield and cell viability of SVF isolated from lipoaspirates were: group A, (7.06±1.28)×10 5/ml and 82.46%±2.81%; group B, (6.90±0.32)×10 5/ml and 82.01%±3.85%; group C, (6.40±0.62)×10 5/ml and 77.82%±3.45%, respectively. No significant difference was found in different age groups. SVF viability was decreased with increasing age. The expression of positive surface markers CD90, CD44, CD105 and CD73 of ASCs in each group was above 95%, and the expression of negative surface markers was below 2%, all of which met the criteria for the expression level of mesenchymal stem cell surface markers. Moreover, there was a decline in cell proliferation and migration of ASCs with increasing age. No significant difference was found in the adipogenic differentiation of ASCs in three groups. The fat grafts were harvested three months after cell-assisted lipotransfer. The graft weight was(0.18±0.02) g in group A, (0.17±0.02) g in group B, (0.15±0.01) g in group C, (0.13±0.03) g in control group, respectively; F=9.274, P<0.001. The residual volume of grafts was(262.88±17.69)/mm 3 in group A, (263.83±25.96)/mm 3 in group B, (240.06±25.08)/mm 3 in group C, (201.81±31.48)/mm 3 in the control group; F=12.95, P<0.001. There were significant differences in the weight and residual volume of fat grafts in different age groups( F=5.231, P=0.012; F=3.364, P=0.049). HE staining result showed that compared with the blank control group, ASC-assisted groups had uniform distribution of adipocytes, less fibrous connective tissue and necrotic tissue. There was a statistically significant difference in the proportion of fat integrity and necrotic tissue between the groups ( F=3.434, P=0.027; F=9.314, P<0.001). Results of the histologic analysis showed no significant difference in the proportion of fat cell integrity and necrotic tissue in each group( F=0.282, P=0.756; F=0.421, P=0.661). Immunofluorescence staining result showed that, compared with the control group, a higher number of perilipin-positive adipocytes were observed in ASCs-assisted fat grafting from different age groups, with uniform distribution. The vessel density of fat grafts was (15.70±4.16)/mm 2 in group A, (17.03±8.30)/mm 2 in group B; (16.68±6.71)/mm 2 in group C, (11.50±4.04)/mm 2 in control group; F=3.523, P=0.019. Conclusions:The proliferation and migration of human ASCs decreased with age, but age did not affect the adipogenic differentiation potential of ASCs. ASCs from different ages effectively improved the fat survival of grafts. ASCs-assisted fat grafting was more effective in young people than in elder.

Objective:The aim of this study is to compare the effects of age on the biological properties of adipose-derived stem cells(ASCs) and fat survival of ASC-assisted lipotransfer. To identify the effect of age factors on the biological characteristics of human ASCs and compare the effects of ASCs-assisted subcutaneous fat transplantation/lipotransfer on nude mice at different ages.Methods:Human lipoaspirates were obtained from 30 healthy female patients (aged from 18 to 65 years) acquiring the abdominal liposuction. Samples were divided into three groups according to donor age: group A, 18-29 years; group B, 30-49 years; group C, 50-65 years. Stromal vascular fraction cells were isolated from the harvested adipose tissue using collagenase. The yield and cell viability of SVF were tested using the Muse cell count and viability assay. ASCs were cultured and harvested at the second passage. MSC surface markers of ASCs were examined by the flow cytometry. The cell proliferation of ASCs from different age was determined by the CCK-8 assay. The scratch test was used for assessing the ASCs migration ability. The adipogenic differentiation potential of ASCs was analyzed by induction of lipid formation in vitro. The expression levels of PPAR-γ and CEBP-α genes were detected by RT-PCR assay. The survival of adipocytes in the grafts was analyzed by perilipin-A immunofluorescence staining. The fat survival of ASCs-enriched grafts from different age was measured in animal models. The weight and residual volume of fat grafts were compared in different groups after three months. The histologic analysis was evaluated by cell integrity and necrosis tissue in fat grafts. The vessel density was measured using the CD31 immunohistochemical staining. Data were analyzed by SPSS software version 21.0 with one-way ANOVA to compare the difference of multiple groups. A value of P< 0.05 was considered statistically significant. Results:The yield and cell viability of SVF isolated from lipoaspirates were: group A, (7.06±1.28)×10 5/ml and 82.46%±2.81%; group B, (6.90±0.32)×10 5/ml and 82.01%±3.85%; group C, (6.40±0.62)×10 5/ml and 77.82%±3.45%, respectively. No significant difference was found in different age groups. SVF viability was decreased with increasing age. The expression of positive surface markers CD90, CD44, CD105 and CD73 of ASCs in each group was above 95%, and the expression of negative surface markers was below 2%, all of which met the criteria for the expression level of mesenchymal stem cell surface markers. Moreover, there was a decline in cell proliferation and migration of ASCs with increasing age. No significant difference was found in the adipogenic differentiation of ASCs in three groups. The fat grafts were harvested three months after cell-assisted lipotransfer. The graft weight was(0.18±0.02) g in group A, (0.17±0.02) g in group B, (0.15±0.01) g in group C, (0.13±0.03) g in control group, respectively; F=9.274, P<0.001. The residual volume of grafts was(262.88±17.69)/mm 3 in group A, (263.83±25.96)/mm 3 in group B, (240.06±25.08)/mm 3 in group C, (201.81±31.48)/mm 3 in the control group; F=12.95, P<0.001. There were significant differences in the weight and residual volume of fat grafts in different age groups( F=5.231, P=0.012; F=3.364, P=0.049). HE staining result showed that compared with the blank control group, ASC-assisted groups had uniform distribution of adipocytes, less fibrous connective tissue and necrotic tissue. There was a statistically significant difference in the proportion of fat integrity and necrotic tissue between the groups ( F=3.434, P=0.027; F=9.314, P<0.001). Results of the histologic analysis showed no significant difference in the proportion of fat cell integrity and necrotic tissue in each group( F=0.282, P=0.756; F=0.421, P=0.661). Immunofluorescence staining result showed that, compared with the control group, a higher number of perilipin-positive adipocytes were observed in ASCs-assisted fat grafting from different age groups, with uniform distribution. The vessel density of fat grafts was (15.70±4.16)/mm 2 in group A, (17.03±8.30)/mm 2 in group B; (16.68±6.71)/mm 2 in group C, (11.50±4.04)/mm 2 in control group; F=3.523, P=0.019. Conclusions:The proliferation and migration of human ASCs decreased with age, but age did not affect the adipogenic differentiation potential of ASCs. ASCs from different ages effectively improved the fat survival of grafts. ASCs-assisted fat grafting was more effective in young people than in elder.