The field of gene therapy has been increasingly studied in the last four decades, and its clinical application has become a reality in the last 15 years. Traditional Chinese medicine (TCM), an important component of complementary and alternative medicine, has evolved over thousands of years with its own unique system of theories, diagnostics and therapies. TCM is well-known for its various roles in preventing and treating infectious and chronic diseases, and its usage in other modern clinical practice. However, whether TCM can be applied alongside gene therapy is a topic that has not been systematically examined. Here we provide an overview of TCM theories in relation to gene therapy. We believe that TCM theories are congruent with some principles of gene therapy. TCM-derived drugs may also act as gene therapy vehicles, therapeutic genes, synergistic therapeutic treatments, and as co-administrated drugs to reduce side effects. We also discuss in this review some possible approaches to combine TCM and gene therapy.

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.

Objective:To evaluate the effectiveness of TiRobot-assisted surgery by O-arm navigation for minimally invasive treatment of type C pelvic fractures.Methods:A retrospective case control study was conducted on clinical data of 53 patients with type C pelvic ring fractures treated between January 2018 and September 2019 at Shanghai Tenth People’s Hospital. There were 29 males and 24 females，aged 20-68 years［（40.4 ± 15.5）years］. All were diagnosed with AO type C fractures. A total of 32 patients underwent minimally invasive internal fixation by TiRobot orthopedic system assisted with O-arm navigation（TiRobot group），and 21 patients were managed by minimally invasive internal fixation under the fluoroscopy guidance of the C-arm X-ray machine（control group）. The operation time，intraoperative blood loss，fluoroscopy time of screw and good and excellent rate of screw placement were recorded. One week after surgery，the pain intensity was assessed by visual analog scale（VAS），and the fracture reduction quality by Matta radiographic criteria. At the latest follow-up，the recovery of pelvic function was evaluated by Majeed criteria. Complications were recorded after operation and during the follow-up.Results:All patients were followed up for 12-15 months［（13.2 ± 0.3）months］. The operation time was（85.2 ± 6.9）minutes in TiRobot group and（101.0 ± 6.7）minutes in control group（ P < 0.05）. The intraoperative blood loss showed no significant difference between the two groups（ P > 0.05）. The fluoroscopy time of screw placement was 6（5，7）seconds in TiRobot group，compared to 10（10，15）seconds in control group（ P < 0.05）. In TiRobot group，the screw placement was excellent in 30 patients，good in 1，and fair in 1，with the excellent and good rate of 97%（31/32），while in control group，15 patients showed excellent placement of screws，2 good，and 4 fair，with the excellent and good rate of 76%（17/21）（ P < 0.05）. There was no significant difference in VAS between the two groups at postoperative one week（ P > 0.05）.At the latest follow-up，the pelvic reduction quality based on Matta radiographic criteria and pelvic function using Majeed criteria did not differ significantly between the two groups（ P > 0.05）. No complications occurred after operation and during the follow-up，such as loosening or breakage of screws，heterotopic ossification，vascular injury or nerve injury. Conclusion:For type C pelvic fractures，TiRobot-assisted minimally invasive internal fixation by O-arm navigation can shorten operation time and intraoperative fluoroscopy time，and improve the accuracy of screw placement when compared to C-arm fluoroscopy.

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.

Objective:To investigate the therapeutic effect of robot-assisted percutaneous screw fixation in LC-II pelvic fractures.Methods:A total of 46 patients with Young-Burgess LC-II pelvic fractures, who underwent surgical treatment at Shanghai Tenth People's Hospital from December 2019 to June 2022, were retrospectively analyzed. Among them, 27 were male and 19 were female, with an average age of 39.78±15.11 years (range, 19-68 years). The causes of injury were traffic accidents (28 cases) and fall from height (18 cases); all the cases were Type B pelvic fractures according to the Tile classification. The patients were divided into two groups based on the surgical approach: the robot group (26 cases) treated by Tianji robot-assisted percutaneous screw fixation, and the actual screw group (20 cases) treated by minimally invasive internal fixation under fluoroscopic guidance. The surgical time, blood loss, number of guide wire adjustments, the success rate of initial screw placement, postoperative vertical and horizontal displacement, and postoperative pelvic fracture reduction quality (Matta score) were compared between the two groups. Visual analogue scale (VAS) was used to assess patients' pain levels before and one week after surgery, and the clinical efficacy was evaluated using the Majeed score at the final follow-up. The occurrence of complications such as infection, vascular and nerve injury, and screw breakage was all recorded during the follow-up.Results:All patients successfully underwent the surgery and were followed up for an average of 11.93±3.09 months (range, 5-15 months). A total of 84 percutaneous screws were applied in the 46 cases, with 47 screws in the robot group and 37 screws in the actual screw group. The surgical time and number of guide wire adjustments were 81.27±4.90 minutes and 0.58±0.76 times in the robot group, 102.95±8.73 minutes and 5.80±2.24 times in the actual screw group, with significant differences ( t=-10.68, P<0.001; t=-11.13, P<0.001). There was no significant difference between the two groups in terms of intraoperative blood loss and postoperative vertical and horizontal displacement ( P>0.05). The success rate of initial screw placement in the robot group was 98% (46/47, 44 excellent, 2 good, 1 poor), while in the actual screw group, it was 86% (32/37, 27 excellent, 5 good, 5 poor), with a significant difference (χ 2=4.05, P=0.044). The VAS scores at one week postoperatively were 3.38±1.20 in the robot group and 6.80±1.61 in the actual screw group, with a significant difference ( t=-8.25, P<0.001). The excellent rate of Matta score after surgery was 92% in the robot group and 85% in the actual screw group, and the Majeed scores at the final follow-up were 86.54±3.10 and 85.7±2.20 respectively, with no significant difference between the two groups ( P>0.05). All patients achieved primary healing; no complications such as infection or screw breakage occurred during the follow-up period. Conclusion:Robot-assisted percutaneous access screw fixation for LC-II pelvic fracture can achieve accurate reduction of pelvic fracture, and is characterized by short operative time, low radiation exposure, and high safety and accuracy.

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 retrospectively analyze the long-term survival (10-15 years) and late toxicity of nasopharyngeal carcinoma (NPC) patients after intensity-modulated radiotherapy (IMRT), aiming to provide reference for the optimal treatment of NPC.Methods:132 patients with NPC who were treated with IMRT in Sichuan Cancer Hospital from 2003 to 2009 were recruited. Among them, 3 patients were classified as stage Ⅰ, 22 cases of grade Ⅱ, 61 cases of grade Ⅲ, 43 cases of Ⅳ A and 3 cases of Ⅳ B, respectively. The median dose was 73.37Gy (66 to 85Gy), divided into 33 times. Twenty patients received radiotherapy alone, 112 cases of concurrent radiochemotherapy. The survival rate was calculated by Kaplan-Meier method and log- rank test. Univariate prognostic analysis was performed. Cox model was used to conduct multivariate prognostic analysis. The late radiation toxicity was evaluated by RTOG/EORTC criteria. Results:The median follow-up duration was 128 months (range, 3 to 191 months). The 10-and 15-year local control rates of NPC patients were 86.0% and 79.9%. The disease-free survival rates were 72.5% and 63.2%, and the overall survival (OS) rates were 65.2% and 57.1%. The local recurrence rate was 12.1%, and the distant metastasis rate was 16.7%. A total of 53 patients died, of whom 15 patients died of local recurrence, 20 patients died of distant metastasis and 18 patients died of other diseases (pneumonia, intracranial hemorrhage and accident, etc.). The 10-and 15-year non-tumor-related mortality rates were 11.3% and 13.6%. Univariate analysis showed that age, smoking habit, lactate dehydrogenase (LDH), T stage and clinical stage were the independent prognostic factors of OS in NPC patients. Multivariate analysis demonstrated that LDH, T stage and synchronous chemotherapy were the prognostic factors of OS in NPC patients. The incidence of gradeⅠ-Ⅱ late radiation injury (hearing impairment, dysphagia, dental caries and xerostomia) was 90.4%, and 8.5% for grade Ⅲ-Ⅳ late radiation injury (skin fibrosis, hearing impairment and radiation brain injury).Conclusions:The 10-and 15-year OS of NPC patients treated with IMRT is relatively high. With the prolongation of survival, the non-tumor-related mortality rate is increased. Distant metastasis is the main cause of treatment failure. The main late injuries include grade Ⅰ/Ⅱ hearing impairment, dysphagia, dental caries and xerostomia.

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.

Blood supply is believed to be an important aspect in the development of pathological scars. However, there are controversies about vascular distribution, vascular structure and blood flow in pathological scars. Additionally, hypoxic microenvironment plays an important role in the vascularization of pathological scar tissues, and hypoxic conditions can be reflected by metabolic indexes and some cytokines. Furthermore, the correlation between blood supply and tissue hypoxia is controversial. The aim of this article is to review the literature on the characteristics of blood supply and tissue hypoxia in pathological scars, from which we can see pathological scars have unique characteristics of blood supply that are closely associated with tissue hypoxia. Moreover, development in the treatment of pathological scars is herein reviewed.
Cell Hypoxia ; Cicatrix ; blood ; metabolism ; Humans ; Regional Blood Flow