The Application of "Table Tennis Racquet" Random Skin Flap in the Treatment of Facial Skin Carcinoma.

BACKGROUND: The repair of facial skin and soft tissue defects remains a clinical challenge. The author introduced a novel "table tennis racquet" random skin flap for wound repair after facial skin cancer excision and discussed its survival mechanisms. METHODS: A lateral mandibular neck skin flap shaped like a table tennis racquet with no well-known blood vessels at the narrow pedicle was designed in 31 cases to repair tissue defects. Among them, there were 8 cases of skin carcinoma in the frontotemporal area and 23 cases of skin carcinoma in the cheek. The flap area was 8.0 × 7.0 cm at maximum and 3.0 × 2.5 cm at minimum, with a pedicle width of 1.0-2.0 cm and a pedicle length of 2.0-6.0 cm. RESULTS: All 31 "table tennis racquet" random skin flaps survived, although there were 3 cases with delayed healing of distal flap bruising. All of them had an ideal local shape after repair with a concealed donor area and inconspicuous scars. CONCLUSIONS: This flap has a "table tennis racquet" shape with a pedicle without well-known blood vessels and has a length-to-width ratio that exceeds that of conventional random flaps, making it unconventional. Because of its long and narrow pedicle, it not only has a large rotation and coverage area but also can be designed away from the defect area, avoiding the defect of no donor tissue being localized near the defect. Overall, this approach is an ideal option for repairing tissue defects after enlarged excision of facial skin carcinoma.

A B-box zinc finger protein, MdBBX10, enhanced salt and drought stresses tolerance in Arabidopsis.

KEY MESSAGE: The expression of MdBBX10 was significantly induced by different stresses and ABA treatments. Overexpression of MdBBX10 in Arabidopsis significantly enhanced abiotic stresses tolerance by ABA signalling. The roles of B-box domain(s) containing proteins (BBXs) in regulation of flowering and light morphogenesis of plants were intensively studied. However, the roles of plant BBXs in abiotic stresses are poorly understood. A B-box protein encoding gene from apple (MdBBX10) was found to be up-regulated from gene expression profile under salt stress. qRT-PCR analysis indicated that the expression of MdBBX10 was significantly induced by different stresses and exogenous abscisic acid (ABA) in apple roots and leaves. The ß-glucuronidase activity driven by the promoter of MdBBX10 was also strongly induced by NaCl, H2O2, polyethylene glycol and exogenous ABA, which was consistent to the existence of rich cis-acting elements related to the abiotic stresses in the promoter sequence. Over-expression of MdBBX10 in Arabidopsis significantly enhanced tolerance to abiotic stresses, with higher germination ratio and longer length of roots than the wild type plants. Transgenic plants of over-expressing MdBBX10 lines were more sensitive to exogenous ABA than the wild type plants. Under abiotic stress treatments, the transcript levels of ABA- and stress-related genes were higher in MdBBX10-overexpressing plants than wild type plants. Over-expression of MdBBX10 could enhance plant's ability to scavenge reactive oxygen species (ROS) under stresses, which is correlated with the expression of ROS-scavenging genes. These results provided the evidences that MdBBX10 plays an important role in enhanced plant tolerance to abiotic stresses, which were involved in ABA-mediated response and ROS response.

Polymer-Covalent Organic Frameworks Composites for Glucose and pH Dual-Responsive Insulin Delivery in Mice.

Glucose and pH dual-responsive insulin delivery carriers that have been validated in animal models, remain elusive and much desired. Herein, a new class of covalent organic frameworks (COFs)-based insulin delivery nanocarriers is developed by encapsulating insulin (Ins) and glucose oxidase (GOx) into COFs (COF-1 and COF-5) via both Brønsted and Lewis type (N:→B) complexations. Subsequently, polyethylene glycolated fluorescein isothiocyanate (FITC-PEG) is incorporated into the COFs via the exchange reactions between the disulfide in insulin chains and the thiol in FITC-PEG to afford a robust nano-assembly (FITC-PEG-COF@Ins-GOx). In vitro, the nanocarriers rely on the boroxine-linked COFs' response to pH and glucose dual-stimulation and rendered sustainable insulin delivery. In vivo, the polymer-COFs composite displays excellent long-acting anti-diabetic effects on type 1 diabetic mice within 72 h without side effects after one injection. More intriguingly, the nanocomposites also show great promise for the efficient delivery of native proteins with high generality. To the authors' knowledge, this represents the first study pertaining to a facile methodology to prepare COF-based insulin-delivery nanocarriers for in vitro and in vivo therapeutic applications.

Members of B-box Protein Family from Malus domestica Enhanced Abiotic Stresses Tolerance in Escherichia coli.

The B-box proteins (BBXs) are zinc finger proteins containing one or two B-box domain(s) and involved in regulation of development processes as transcription factors in plants. Here, seven BBX genes in Malus domestica genome (MdBBXs) were identified and found to be up-regulated under abiotic stresses, with 2-12 folds in roots. All recombinant MdBBXs expressed in Escherichia coli (E. coli) enhanced the cell's tolerance to salt and osmotic stresses, respectively. Deficiency of B-box domain of MdBBX10 led to the loss of anti-stress functions. Five conservative cysteines in B-box domain played crucial roles in stress resistance, which are involved in two of metal iron binding sites of zinc finger motifs in BBXs. All the above results suggested MdBBXs confer stress tolerance to E. coli cell against abiotic stresses.

Polyethylenimine and sodium cholate-modified ethosomes complex as multidrug carriers for the treatment of melanoma through transdermal delivery.

Aim: Multidrug resistance is the main reason for the failure of chemotherapy during the treatment of the tumor. To overcome multidrug resistance, this study attempts to develop a novel transdermal drug-delivery system (TDDS) loading cytotoxic drug and chemosensitizer. Materials & methods: The polyethylenimine-modified ethosomes (Eth-PEI) and sodium cholate-modified ethosomes (Eth-SC) were firstly fabricated, and then a novel TDDS based on the carriers complex of Eth-PEI/Eth-SC was prepared by electrostatic interaction and evaluated both in vitro and in vivo. Results: The Eth-PEI/Eth-SC showed the excellent antitumor effect on treating melanoma, using doxorubicin and curcumin as the cytotoxic drug and chemosensitizer, respectively. Conclusion: The as-prepared TDDS composed of Eth-PEI/Eth-SC loading multidrug is an effective means for treating melanoma.

PvNAC1 increases biomass and enhances salt tolerance by decreasing Na+ accumulation and promoting ROS scavenging in switchgrass (Panicum virgatum L.).

Switchgrass (Panicum virgatum L.) is a bioenergy crop; thus, it is important to improve biomass to effectively produce bioethanol, particularly under adverse stress conditions. NAC transcription factors are involved in the abiotic stress response. PvNAC1 was isolated in the nucleus of switchgrass, with its C-terminal region containing a transcriptional activation domain. PvNAC1 expression was induced by dehydration, salt, H2O2, and abscisic acid treatments. Overexpressing (OE) PvNAC1 improved growth performance, leading to significantly taller and heavier (dry weight) plants. Moreover, cellulose content was significantly higher in OE plants, indicating that PvNAC1 plays an important role regulating growth and bioethanol production. PvNAC1 RNA interference (RNAi) switchgrass plants exhibited reduced dry weight and cellulose content. OE PvNAC1 enhanced tolerance to salt stress, through higher reactive oxygen species scavenging ability and less Na+ and more K+ accumulation in roots and shoots. RNAi plants were more sensitive to salt stress. The quantitative polymerase chain reaction results revealed that some stress responsive genes, three antioxidant enzymatic genes, and an ion homeostasis-related gene were upregulated in OE plants and downregulated in RNAi plants. These results show that PvNAC1 functions as a transcriptional activator in response to salt stress and growth.

[Androgen-secreting tissue construction with co-cultured rat testes somatic cells by tissue engineering technique].

OBJECTIVE: To explore the feasibility of constructing androgen-secreting tissue of a certain size and shape using co-cultured somatic cells of rat testis. METHODS: Thirty male Wistar rats were castrated. model and implanted rat model were prepared by resecting bilateral testes. The suspension of mixed testes cells was cultured to obtain various somatic cells of testes and Leydig cells were collected by differential anchorage-dependent method. These two kinds of cells were seeded onto biodegradable scaffolds of polyglycolic acid (PGA) fibers and cultured in vitro. The tissue formation of cell-scaffold constructs was observed by optical microscope and electronic microscope and the level of testosterone in the supernatant was detected regularly. After 7-day culture in vitro, the 2 kinds of cell-scaffold constructs, scaffold with purified Leydig cells or co-cultured testis somatic cells (seed cells), were implanted into the gastrocolic omentum or cavity of tunica vaginalis of the castrated rats. The implants were harvested 4, 6, 9, 12, and 24 weeks later to evaluate the tissue formation of cell-scaffold constructs in vivo. The serum testosterone level of the implanted rats was assayed to evaluate the testosterone secreting function of the regenerative tissue. RESULTS: Both the co-cultured testis somatic cells and Leydig cells had fine compatibility with the PGA fibers and adhered to the scaffolds very well. Testosterone was detected at a certain degree in the supernatant of cell-scaffold constructs, indicating the testosterone secreting function of the constructs. Two months after the implantation both kinds of cell-scaffold constructs formed testosterone secreting tissue in both gastrocolic omentum and cavity of tunica vaginalis of the implanted rats. The regenerative tissues were vascularized very well with a certain size and shape. Six weeks after implantation the serum testosterone level of the Leydig cell group was 0.60 ng/ml +/- 0.04 ng/ml, and that of the co-culture group was 0.84 ng/ml +/- 0.03 ng/ml, both significantly higher than that of the control castrated rats (0.56 ng/ml +/- 0.05 ng/ml, both P < 0.01), and the serum testosterone level of the co-cultured testes somatic cell implantation group was significantly higher than that of the Leydig cell implantation group too (P < 0.01). CONCLUSION: It is completely feasible to construct androgen-secreting tissue in vitro and in vivo using tissue engineering technique. Co-cultured testis somatic cells may serve as the better seed cells for androgen-secreting tissue engineering than purified Leydig cells in terms of the quantity and function of cells.

Repair of osteochondral defects with in vitro engineered cartilage based on autologous bone marrow stromal cells in a swine model.

Functional reconstruction of large osteochondral defects is always a major challenge in articular surgery. Some studies have reported the feasibility of repairing articular osteochondral defects using bone marrow stromal cells (BMSCs) and biodegradable scaffolds. However, no significant breakthroughs have been achieved in clinical translation due to the instability of in vivo cartilage regeneration based on direct cell-scaffold construct implantation. To overcome the disadvantages of direct cell-scaffold construct implantation, the current study proposed an in vitro cartilage regeneration strategy, providing relatively mature cartilage-like tissue with superior mechanical properties. Our strategy involved in vitro cartilage engineering, repair of osteochondral defects, and evaluation of in vivo repair efficacy. The results demonstrated that BMSC engineered cartilage in vitro (BEC-vitro) presented a time-depended maturation process. The implantation of BEC-vitro alone could successfully realize tissue-specific repair of osteochondral defects with both cartilage and subchondral bone. Furthermore, the maturity level of BEC-vitro had significant influence on the repaired results. These results indicated that in vitro cartilage regeneration using BMSCs is a promising strategy for functional reconstruction of osteochondral defect, thus promoting the clinical translation of cartilage regeneration techniques incorporating BMSCs.

Repair of porcine articular osteochondral defects in non-weightbearing areas with autologous bone marrow stromal cells.

In vivo niche is known to play important roles in terminal differentiation of implanted bone marrow stromal cells (BMSCs). This study explored the feasibility of repairing articular osteochondral defects using autologous BMSCs and biodegradable polymers. BMSCs from 18 hybrid pigs' marrows were either treated with dexamethasone (40 ng/mL) alone or chondrogenically induced with dexamethasone and transforming growth factor-beta1 (10 ng/mL). The cells were seeded respectively onto polylactic acid (PLA)- coated polyglycolic acid (PGA) scaffolds. Four osteochondral defects in each animal were created at non-weightbearing areas of knee joints (2/each side) and were respectively repaired by a chondrogenically induced BMSC-PGA/PLA construct in experimental group (Exp), by a dexamethasone-treated BMSC-PGA/PLA construct in control 1 group (Ctrl 1), by a PGA/PLA construct alone in control 2 group (Ctrl 2), or left unrepaired in control 3 group (Ctrl 3). To trace the implanted cells, green fluorescent protein (GFP)- labeled BMSCs were implanted in 2 animals. Gross view and histology showed that Exp and Ctrl 1 (with cell implantation) achieved better reparative results than Ctrl 2 and Ctrl 3 (without cell implantation) in terms of the reparative level and the restoration of the histological structure. In addition, 6-month results were better than 3-month results in all 4 groups. In Exp, 11 of 16 defects were completely repaired by hyaline cartilage and cancellous bone. In Ctrl 1, 11 of 16 defects were repaired by fibrocartilage and cancellous bone, although the repair with hyaline cartilage and cancellous bone was observed in 5 of 16 defects. In contrast, no obvious repair or only fibrotic tissue was observed in Ctrl 2 and Ctrl 3. The compressive moduli of repaired cartilage in Exp reached 80.27% of the normal amount at 6 months, with a high level of glycosaminoglycan (GAG) content (no statistical difference from normal). In Ctrl 1, the compressive moduli and GAG content were 62.69% and 78.03% of normal levels, respectively. More importantly, GFP-labeled cells were detected in the engineered cartilage and the repaired subchondral bone. These results strongly indicate that the implanted BMSCs can differentiate into either chondrocytes or osteoblasts and repair articular osteochondral defects by forming engineered cartilage and engineered bone.

Temporal and spatial CGRP innervation in recombinant human bone morphogenetic protein induced spinal fusion in rabbits.

STUDY DESIGN: Animal experiment using a rabbit posterolateral intertransverse process fusion model. OBJECTIVE: To explore the temporal and spatial distribution of sensory nerve fibers expressing calcitonin-gene related peptide (CGRP) during spinal fusion induced by recombinant human bone morphogenetic protein-4 and the role of the CGRP innervation in ectopic bone formation and remodeling. SUMMARY OF BACKGROUND DATA: Sensory neuropeptide CGRP involved in local bone turnover has been evidenced but its underlying mechanism is poorly understood. Knowledge in the CGRP innervation in ectopic bone induced by bone morphogenetic proteins can help us to understand its role in bone turnover. METHODS: Twenty-seven New Zealand white rabbits underwent single level posterolateral intertransverse process fusion of the lumbar vertebrae with implantation of porous poly-d,l-lactic acid blocks loaded with 1.25 microg recombinant human bone morphogenetic protein-4 solution. Animals were killed and the operated lumbar vertebrae were harvested for histomorphological evaluation at 3 days (n = 3), 1 week (n = 6), 3 weeks (n = 6), 7 weeks (n = 6), and 12 weeks (n = 6) following surgery, respectively. RESULTS: New cartilage presented at 1 week postimplantation adjacent to the implant, reached a peak volume at week 3 followed by a drop till week 12 after its ossification. Trabeculae-like woven bone structure presented at week 3. CGRP-positive nerve fibers regenerated already at 3 days postimplantation, reached its peak density at week 3. The CGRP-positive fibers presented both in fibrous tissues adjacent to proliferating cartilages and in bone marrow of newly formed trabecular bone. CONCLUSIONS: The observed spatial and temporal regeneration of CGRP-positive nerve fibers in ectopic bone formation suggested CGRP innervation is associated with ectopic osteogenesis.