Identification and Validation of a m5C RNA Modification-Related Gene Signature for Predicting Prognosis and Immunotherapeutic Efficiency of Gastric Cancer.

Background: 5-methylcytosine (m5C) is a major site of RNA methylation modification, and its abnormal modification is associated with the development of gastric cancer (GC). This study aimed to explore the value of m5C-related genes on the prognosis of GC patients through bioinformatics. Methods: First, m5C-related genes were obtained by nonnegative matrix factorization (NMF) analysis and differentially expressed analysis. The m5C-related model was established and validated in distinct datasets. Moreover, a differential analysis of risk scores according to clinical characteristics was performed. The enrichment analysis was carried out to elucidate the underlying molecular mechanisms. Furthermore, we calculated the differences in immunotherapy and chemotherapy sensitivity between the high- and low-risk groups. Finally, we validated the expression levels of identified model genes by quantitative real-time polymerase chain reaction (qRT-PCR). Results: A total of five m5C-related subtypes of GC patients in the TCGA database were identified. The m5C-related model was constructed based on APOD, ASCL2, MFAP2, and CREB3L3. Functional enrichment revealed that the m5C-related model might involve in the cell cycle and cell adhesion. Moreover, the high-risk group had a higher abundance of stromal and immune cells in malignant tumor tissues and a lower tumor purity than the low-risk group. The patients in the high-risk group were more sensitive to chemotherapy and had better sensitivity to CTLA4 inhibitors. Furthermore, qRT-PCR results from our specimens verified an over-expression of ASCL2, CREB3L3, and MFAP2 in the cancer cells compared with the normal cells. Conclusion: A total of five GC subtypes were identified, and a risk model was constructed based on m5C modification.

MiR146a-loaded engineered exosomes released from silk fibroin patch promote diabetic wound healing by targeting IRAK1.

Unhealable diabetic wounds need to be addressed with the help of newer, more efficacious strategies. Exosomes combined with biomaterials for sustained delivery of therapeutic agents are expected to bring new hope for chronic wound treatment. Here, the engineered exosomes modified for efficiently loading miR146a and attaching to silk fibroin patch (SFP) were demonstrated to promote diabetic wound healing. Silk fibroin binding peptide (SFBP) was screened through phage display, and SFBP-Gluc-MS2 (SGM) and pac-miR146a-pac fusion protein were constructed. The designed exosomes (SGM-Exos, miR146a-Exos, and SGM-miR146a-Exos) were isolated from the engineered placental mesenchymal stem cells (PMSCs) transduced with SGM or/and pac-miR146a-pac protein. Gluc signals indicated SGM-Exo@SFP markedly increased the binding rate and the stability of SGM-Exo. Moreover, the loading efficiency of miR146a in SGM-miR146a-Exos was ten-fold higher than that in miR146a-Exos. Superior to untreated, SGM-miR146a-Exo-only treated, and SFP-only treated groups, SGM-miR146a-Exo@SFP drived wound healing associated with less inflammation, collagen deposition, and neovascularization. The transcriptomics analysis suggested anti-inflammatory and regenerative effects with SGM-miR146a-Exo@SFP treatment. Here, we show efficient exosome@biomaterial-based miRNA delivery systems for regenerative medicine and tissue engineering.

Sodium alginate/gelatin hydrogels loaded with adipose-derived mesenchymal stem cells promote wound healing in diabetic rats.

BACKGROUND: Chronic refractory wounds are a common complication in diabetic patients. Adipose-derived mesenchymal stem cells (ASCs) have been shown to play an essential role in diabetic wound repair. AIMS: To determine whether a composite of ASCs and sodium alginate/gelatin (Gel-Al) hydrogel can promote diabetic wound healing. METHODS: Full-thickness cutaneous wounds were created in streptozotocin-induced diabetic rats prior to treatment with Gel-Al hydrogels loaded with ASCs. Hydrogel biocompatibility and wound healing were analyzed. Hematoxylin and eosin staining, Masson staining, immunofluorescence, enzyme-linked immunosorbent assays (ELISA), and quantitative real-time PCR were performed for the assessment of cellular responses. RESULTS: Compared to the control group or Gel-Al alone group, the combination of Gel-Al and ASCs promoted wound closure, facilitated granulation tissue regeneration and collagen deposition, and upregulated the expression of vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), epidermal growth factor (EGF), and endothelial cell marker CD31. Moreover, the combination of Gel-Al and ASCs decreased interleukin-6 (IL-6) and interleukin-1ß (IL-1ß) expression, increased transforming growth factor beta1 (TGFß1), interleukin-10 (IL-10), interleukin-4 (IL-4) and interleukin-13 (IL-13) expression, and increased M2 macrophage polarization. CONCLUSIONS: Gel-Al hydrogels loaded with ASCs accelerate diabetic wound healing. The Gel-Al hydrogel-based ASC system therefore represents an innovative therapeutic strategy for diabetic wound repair.

Cancer Progression Mediated by CAFs Relating to HCC and Identification of Genetic Characteristics Influencing Prognosis.

Background: Hepatocellular carcinoma (HCC) is one of the most common malignancies, and although there are several treatment options, the overall results are not satisfactory. Cancer-associated fibroblasts (CAFs) can promote cancer progression through various mechanisms. Methods: HCC-associated mRNA data were sourced from The Cancer Genome Atlas database (TCGA) and International Cancer Genome Consortium (ICGC) database. First, the differentially expressed CAF-related genes (CAF-DEGs) were acquired by difference analysis and weighted gene coexpression network analysis (WGCNA). Moreover, a CAF-related risk model was built by Cox analysis. Kaplan-Meier (K-M) curves and receiver operating characteristic (ROC) curves were utilized to evaluate the validity of this risk model. Furthermore, enrichment analysis of differentially expressed genes (DEGs) between the high- and low-risk groups was executed to explore the functions relevant to the risk model. Furthermore, this study compared the differences in immune infiltration, immunotherapy, and drug sensitivity between the high- and low-risk groups. Finally, we verified the mRNA expression levels of selected prognostic genes by quantitative real-time polymerase chain reaction (qRT-PCR). Results: 107 CAF-DEGs were identified in the HCC samples, and five prognosis-related genes (ACTA2, IGJ, CTHRC1, CXCL12, and LAMB1) were obtained by Cox analysis and utilized to build a CAF-related risk model. K-M analysis illustrated a low survival in the high-risk group, and ROC curves revealed that the risk model could accurately predict the 1-, 3-, and 5-year overall survival (OS) of HCC patients. In addition, Cox analysis demonstrated that the risk score was an independent prognostic factor. Enrichment analysis illustrated that DEGs between the high- and low-risk groups were related to immune response, amino acid metabolism, and fatty acid metabolism. Furthermore, risk scores were correlated with the tumor microenvironment, CAF scores, and TIDE scores, and CAF-related marker genes were positively correlated with all five model genes. Notably, the risk model was relevant to the sensitivity of chemotherapy drugs. Finally, the results of qRT-PCR demonstrated that the expression levels of 5 model genes were in accordance with the analysis. Conclusion: A CAF-related risk model based on ACTA2, IGJ, CTHRC1, CXCL12, and LAMB1 was built and could be utilized to predict the prognosis and treatment of HCC.

Promotive effects of four herbal medicine ARCC on wound healing in mice and human.

Background: Traditional Chinese medicine (TCM) had been extensively used in China for wound management and had shown great potential in wound treatment while its mechanism is still needed to be addressed. Objective: The present study sought to investigate the therapuetic effect of the TCM ARCC on acute and chronic wounds. Methods: Here, using the ultra-low temperature preparation method, the mixed ultramicro powder prepared with Angelica (A), Angelica (R), Calcined Gypsum (C) and Caleramide (C) named as ARCC. The effects of ARCC on wound healing in adult and aged mice were comparatively evaluated through a full-thickness skin defect model. In addition, we randomly selected 10 patients aged 55 to 70 years from a cohort of 500 patients with diabetic feet to assess their prognosis. Results: As the results showed that the healing rate had delayed in aged mice compared to adult mice, while ARCC prominently augmented the healing process in aged mice. Moreover, ARCC treatment wounds in aged mice showed accelerated re-epithelization, enhanced granulation tissue formation, and increased vascularization, which was similar to that of adult mice. Furthermore, ARCC also achieved therapeutic effects in diabetic foot patients, accelerating wound healing. The results found that foot ulcers improved significantly 7 days after the ARCC administration, and 80% of patients were healed within 1 month. Discussion: In the present study, ARCC was found to have therapeutic effects on both acute and chronic wounds in animal models. ARCC also demonstrated therapeutic effects in diabetic feet, which promoted wound healing, prevented wound infection, and avoided the risk of further surgery or amputation. All these evidences suggested ARCC was a promising approach for wound treatment. Conclusions: ARCC might be recommended as a promising therapeutic medication in diabetic and chronic refractory wounds.

[Research progress of chronic wound debridement].

Objective: To review the research progress of chronic wound debridement. Methods: The recent related literature concerning the mechanisms, advantages, limitations, and indications of the technologies of chronic wound debridement was extensively consulted, reviewed, and summarized. Results: Debridement is essential for chronic wound healing, which includes autolytic debridement, enzymatic debridement, biodebridement, mechanical debridement, sharp/surgical debridement, ultrasound debridement, hydrosurgery debridement, and coblation debridement. Each technique has its own advantages and disadvantages. Conclusion: There are many types of technologies of chronic wound debridement, which can be chosen according to clinical conditions. It is showed there are more significant advantages associated with the technique of coblation debridement relatively, which also has greater potential. Further study is needed to improve its efficacy.

[Application of plasma-mediated bipolar radiofrequency ablation debridement in treatment with retention of internal fixation for early postoperative infection of fractures of extremities].

Objective: To explore the effectiveness of plasma-mediated bipolar radiofrequency ablation debridement (Coblation debridement) in treatment with retention of internal fixation for early postoperative infection of fractures of extremities. Methods: Between January 2012 and May 2015, 16 patients (12 males and 4 females) with early postoperative infection of internal fixation for extremity fracture were treated, with an average age of 41.6 years (range, 19-61 years). The fractures included tibia and fibula fracture in 5 cases, femoral fractures in 5 cases, distal humeral fractures in 3 cases, ulna and radius fractures in 2 cases, and patellar fracture in 1 case. Two cases were open fractures and 14 cases were closed fractures. All fractures were fixed non-intramedullarily. Postoperative infection occurred at 5-10 days (mean, 7 days) after operation, with bacteria cultured from wound secretion in all cases. Type EIC5872 70 Coblation knife and Coblator Ⅱ plasma surgery system were conducted to debride the foci of infection in ablating pattern with sterile saline used as the conductive fluid and the magnitude of power from 6 to 9. After pulse irrigating with sterile saline, irrigating tube and draining tube were placed beside the fixation. Postoperative continuous irrigation and drainage and systemic antibiotic therapy would be conducted. The effectiveness was evaluated by bone infection effectiveness evaluation criteria. Results: All the 16 patients were followed up 12-36 months (mean, 15 months) after operation. All the infected wounds were cured and healed by first intention without recurrence in all the patients, and the fracture healing time was 3-7 months (mean, 4.8 months) without limb dysfunction or nonunion. Internal fixation was removed at 1-2 years after operation in 4 cases, whom with good fracture healing and without recurrence of infection after operation. Conclusion: The effectiveness of Coblation debridement in treatment with retention of internal fixation for early postoperative infection of extremity fractures are satisfactory, which can avoid the second stage operation, infectious nonunion, and osteomyelitis.

[Experimental study on radiofrequency ablation technology for treatment of infected wounds in minipigs].

OBJECTIVE: To observe the effect of radiofrequency ablation technology for the treatment of infected wounds in minipigs. METHODS: Infected wounds of full-thickness skin defects (about 6.15 cm2/wound) were prepared in 8 6-month-old minipigs (weighing, 30-35 kg) using the method of Davis et al. The 160 wounds were randomly divided into 4 groups (n = 40). Infected wounds were debrided with the radiofrequency ablation technology in group A, with the electric knife in group B, and with the scalpel in group C; no treatment was done in group D as a control. The healing rate, healing time, and tissue filling rate were observed; bacterial quantitative examination and histological examination were done at 0, 2, 7, and 14 days after operation. RESULTS: All infected wounds were successfully established after 48 hours when Staphylococcus aureus dilution were inoculated. The wounds after radiofrequency ablation technology treatment were fresh and flat with slight bleeding; the healing time of group A was significantly shorter than that of groups B, C, and D (P < 0.05), and the healing rate of group A was significantly higher than that of groups B, C, and D at 7 and 14 days after operation (P < 0.05). The tissue filling rate of group A was significantly higher than that of groups B, C, and D at 2 days after operation (P < 0.05); the tissue filling rates of groups A, B, and C were significantly higher than that of group D at 7 and 14 days after operation (P < 0.05). At 0, 2, 7, and 14 days, there were significant differences in the bacterial count per gram tissue among 4 groups (P < 0.05), the order from low to high was groups A, B, C, and D. The histological observation showed that the surface of wound was smooth in group A at 0 day, and group A was better than the other groups in wound healing; at 2 days, some exudates were observed in 4 groups, but it was least in group A. There was inflammatory cell infiltration in various degrees in 4 groups at 7 and 14 days; it was lightest in group A with thick epithelium and dense collagen bundles, followed by groups B and C, and it was severe in group D. CONCLUSION: The radiofrequency ablation technology can effectively remove the necrotic tissues of infected wounds, remarkably reduce the number of bacteria, improve the healing rate, and shorten the healing time of wounds.

[Clinical safety about repairing the peripheral nerve defects with chemically extracted acellular nerve allograft].

OBJECTIVE: To discuss the clinical safety about repairing the peripheral nerve defects with the acellular allogeneic nerve. METHODS: The 41 patients (male 38, female 3, age 10 - 55 years old, average 28.9 years old) who were performed chemically extracted acellular nerve allograft transplanting to repair nerve defects from 2002 to 2011. The average interval from injury to nerve repairing was 4.1 months (range, 10 hours to 9 months). There were 41 cases nerve defects including 10 brachial plexus nerves, 3 radial nerves of upper arm, 4 ulnar nerves of forearm, 12 digital and toe nerves, 2 sciatic nerves, 2 femoral nerves, 3 tibial nerves and 5 common peroneal nerves. There were 12 cases combined fractures and 20 soft tissue injury or defects. The average length of the nerve allograft to bridge the nerve defects was 6.1 cm (range, 2 - 10 cm). No immunosuppressive drugs were used in all cases. The clinical safety was evaluated through physical examination, blood biochemistry and immunity detection. RESULTS: All cases were followed up post-operation. They got primary wound healing except 2 superficial infection who got delay healing through dressings changing. No any adverse effects happened including immunological rejection, hypersensitivity reaction, deep infection, hepatotoxicity and nephrotoxicity. CONCLUSIONS: It is safe and feasible to repairing human peripheral nerve defects with chemically extracted acellular nerve allograft.

[The application of external fixator for complex tissue defect in the forearm].

OBJECTIVE: To discuss the curative effect of the external fixator for complex tissue defect in the forearm. METHODS: From May, 2005 through December, 2008, the external fixators were used in 17 patients to treat the complex tissue defect in the forearm caused by trauma. There were 11 male and 6 female, with a mean age of 25.6. All patients were accompanied with the exposure of tendon, muscle or screw. The skin defect ranged from 7 cm x4 cm to 19 cm x9 cm. All patients underwent pedicle flap repair. The flap ranged from 10 cm x 6 cm to 20 cm x 15 cm. The proximal pedicle of the flap was sutured into a tubular. The position of the pedicle was fixed by the external fixator. The pin was at the ulnar and the iliac (n=5), and the radius and the iliac (n=12). The immobilization lasted 3 to 8 weeks, 5.1 weeks in average. RESULTS: All patients were followed up for 3 to 20 months, 11.3 in average. All pedicle flaps survived with no pressure ulcer, or no erosion in the axilla. No compartment syndrome or osteomyelitis occurred. Four to six week after surgery, the pedicle was cut. Infection occurred at the cutting end in 1 patient. The wound healed after addressing. The wound in the other 16 patients healed successfully. The fracture of the ulnar and the radius healed 8.5 or 15 weeks after surgery, 13.5 weeks in average. Eleven patients underwent second stage reshape and function restoration. The function of the hands and forearms recovered satisfactorily. Eleven patients returned to their work. Six patients can live with basic function for living. CONCLUSIONS: The external fixator used for complex tissue defect in the forearm can keep the position of the pedicle, replacing plaster fixation. It can reduce the incidence of flap and vessel spasm, and get good outcomes.

[Application of vacuum sealing drainage to treat late-stage large skin avulsion injury with infection].

OBJECTIVE: To investigate the clinical effect of vacuum sealing drainage (VSD) on late-stage large skin avulsion injury with infection. METHODS: From May 2007 to August 2008, 9 patients with large-area skin avulsion injury and infection were treated. There were 1 male and 8 females aged 9-52 years old (median 27 years old). All patients suffered from closed skin avulsion injury involving the lower back, buttock, and part of the thigh. The injury area varied from 30 cm x 25 cm to 92 cm x 38 cm. The time between injury and hospital admission was 15-23 days. The skin avulsion injury was complicated with pelvis fracture, urethral injury, anal injury, sacrum exposure, and limb fractures. The interval between hospital admission and operation was 3-23 hours. Free split-thickness skin graft was performed after the focus debridement and three VSD treatments (40-60 kPa). RESULTS: After three VSD treatments, no patient had general pyemia and severe local tissue necrosis or infection, the tissue edema in the skin avulsion area was alleviated obviously, and all the wound cavities were closed. All the wounds in the graft site healed after 28-45 days of treatment (average 39 days), and all the donor sites healed. Nine patients were followed up for 4-14 months (average 10 months). The appearance of the reparative area was good, and there was no occurrence of joint dysfunction in the injured area due to scar contracture. CONCLUSION: VSD is effective in treating late-stage large skin avulsion injury with infection.

[Transplantation of large anterior latissimus dorsi muscular flap combination with musculus rectus abdominis flap].

OBJECTIVE: To study the value and surgical techniques of transplantation of large anterior latissimus dorsi muscular flap combination with musculus rectus abdominis flap. METHODS: Three cases (2 male and 1 female) with skin defect and bone exposed were reviewed from May 2005 to January 2007. Two patients suffered from trauma, and 1 suffered from tumor resection. Flaps size were: 60 cm x 32 cm, 55 cm x 30 cm and 50 cm x 25 cm, flaps pattern including: 1 free flap with 2 ends of vascular, 1 flap with pedicle and free vascular end, 1 flap with 2 ends of pedicle. RESULTS: Two flaps survived completely, 1 flap with necrosis edge eventually healed after change of dressing. The infection had been effectively controlled and ready for function recovered. One case caused by trauma recovered with fracture healing, full weight-bearing and restore the original work. CONCLUSIONS: Large anterior latissimus dorsi muscular flap combination with musculus rectus abdominis flap can be used for repair of large skin defect. For the difficulty and technical requirements, surgical indications should be strictly controlled.