[Application of Digital Therapy in Post-operative Rehabilitation of Knee Anterior Cruciate Ligament Reconstruction].

OBJECTIVE: Digital therapy is important in treating motor system disease. The outcome of digital therapy in post-operative rehabilitation of knee anterior cruciate ligament (ACL) reconstruction is assessed. METHODS: 142 patients are treated with digital rehabilitation therapy after ACL reconstruction. Patients' pain score, joint motion, lower limb function score, anxiety score are statistically analyzed. Patients' satisfaction, device usage and adverse events are documented. RESULTS: At post-operative 1st day, 8th weeks, 12th weeks, pain score are 4, 2, 1, knee joint range of motion are 55°, 110°, 143°, lower limb function score are 18, 56, 76, anxiety score are 32.5, 26, 23.5 respectively. Patients' satisfaction are 9.4. Mean duration of device usage is (177.6±38.0) minutes per week. Rehabilitation-related and device-related adverse event does not happen. CONCLUSIONS: Digital therapy promotes post-operative rehabilitation after ACL reconstruction.

Role of Arterial Perfusion in Early Survival of Arterialized Venous Flaps: A Mechanism Study and Clinical Application.

BACKGROUND: The arterialized venous flap (AVF) is appropriate as a flap for hand and foot resurfacing meet the aesthetic demands in the same time. However, the inconsistency of survival rate limited its popularization in clinical settings. The purpose of this study was to investigate the role played by the caliber and location of the artery. METHODS: Arterialized venous flaps were designed on the abdomen of New Zealand rabbits, and the animals were randomized into 3 groups and 2 groups in experiment 1 and 2, respectively. In experiment 1, the artery flow was restricted with vascular staplers of different calibers. In experiment 2, the artery was anastomosed with the afferent vein in the center or at the margin of the flap. Blood perfusion state, water content, epidermal metabolite levels, and flap survival status were observed in both experiments. Furthermore, outcomes of 12 patients received AVF to resurface soft tissue defects in the digits, hands, and feet between January 2016 and February 2018 were analyzed. RESULTS: In experiment 1, compared with the control group, groups with restricted artery showed poor results regarding blood perfusion state, water content, epidermal metabolite levels, and flap survival status. In experiment 2, group with the afferent vein in the center of the flap showed better results mentioned previously. All the flaps survived uneventfully in this study. Two flaps partially failed (20% of the flap area) because of insufficient perfusion. Generally, larger caliber and center-located vein helped the survival of AVF. CONCLUSIONS: Experimental findings suggested that increased arterial perfusion and center-located vein are beneficial for the survival of AVF. Clinical series proved the findings previously. The problem of inconsistency of AVF can be partially solved by increasing arterial perfusion and dissecting afferent vein into the center of flap, and still, further studies are needed to shed light on the mechanism behind.

Nkx2-5 Regulates the Proliferation and Migration of H9c2 Cells.

BACKGROUND The protein NKX2-5 affects mammalian heart development. In mice, the disruption of Nkx2-5 has been associated with arrhythmias, abnormal myocardial contraction, abnormal cardiac morphogenesis, and death. However, the details of the mechanisms are unclear. This study was designed to investigate them. MATERIAL AND METHODS Rat cardiomyocytes from the H9c2 cell line were used in our study. First, we knocked down Nkx2-5 in the H9c2 cells and then validated consequent changes in cell proliferation and migration. We then used RNA sequencing to determine the changes in transcripts. Finally, we validated these results by quantitative reverse transcription-polymerase chain reaction. RESULTS We confirmed that Nkx2-5 regulates the proliferation and migration of H9c2 cells. In our experiments, Nkx2-5 regulated the expression of genes related to proliferation, migration, heart development, and disease. Based on bioinformatics analysis, knockdown of Nkx2-5 caused differential expression of genes involved in cardiac development, calcium ion-related biological activity, the transforming growth factor (TGF)-ß signaling pathway, pathways related to heart diseases, the MAPK signaling pathway, and other biological processes and signaling pathways. CONCLUSIONS Nkx2-5 may regulate proliferation and migration of the H9c2 cells through the genes Tgfb-2, Bmp10, Id2, Wt1, Hey1, and Cacna1g; rno-miR-1-3p; the TGF­ß signaling pathway; the MAPK signaling pathway; as well as other genes and pathways.

Analysis of NKX2-5 in 439 Chinese Patients with Sporadic Atrial Septal Defect.

BACKGROUND The NKX2 gene family is made up of core transcription factors that are involved in the morphogenesis of the vertebrate heart. NKx2-5 plays a pivotal role in mouse cardiogenesis, and mutations in NKx2-5 result in an abnormal structure and function of the heart, including atrial septal defect and cardiac electrophysiological abnormalities. MATERIAL AND METHODS To investigate the genetic variation of NKX2-5 in Chinese patients with sporadic atrial septal defect, we sequenced the full length of the NKX2-5 gene in the participants of the study. Four hundred thirty-nine patients and 567 healthy unrelated individuals were recruited. Genomic DNA was extracted from the peripheral blood leukocytes of the participants. DNA samples from the participants were amplified by multiplex PCR and sequenced on an Illumina HiSeq platform. Variations were detected by comparison with a standard reference genome and annotation with a variant effect predictor. RESULTS Thirty variations were detected in Chinese patients with sporadic atrial septal defect, and 6 single nucleotide polymorphisms (SNPs) had a frequency greater than 1%. Among the 30 variations, the SNPs rs2277923 and rs3729753 were extremely prominent, with a high frequency and odds ratio in patients. CONCLUSIONS Single nucleotide variations are the prominent genetic variations of NKX2-5 in Chinese patients with sporadic atrial septal defect. The SNPs rs2277923 and rs3729753 are prominent single nucleotide variations (SNVs) in Chinese patients with sporadic atrial septal defect.

Fisetin inhibits Listeria monocytogenes virulence by interfering with the oligomerization of listeriolysin O.

Listeriolysin O (LLO), an essential virulence determinant of Listeria monocytogenes, is a pore-forming toxin whose primary function is to facilitate cytosolic bacterial replication by breaching the phagosomal membranes, which is critical for the pathogen to evade host immune recognition. The critical role of LLO in the virulence of L. monocytogenes renders it an ideal target for designing novel antivirulence therapeutics. We found that fisetin, a natural flavonoid without antimicrobial activity, is a potent antagonist of LLO-mediated hemolysis. Fisetin effectively inhibits L. monocytogenes infection in both tissue culture and animal infection models. Molecular modeling and mutational analysis revealed that fisetin directly engages loop 2 and loop 3 of LLO, leading to the blockage of cholesterol binding and the reduction of its oligomerization, thus inhibiting its hemolytic activity. Our results establish fisetin as an effective antitoxin agent for LLO, which can be further developed into novel therapeutics against infections caused by L. monocytogenes.

Novel insights into chloroplast genome evolution in the green macroalgal genus Ulva (Ulvophyceae, Chlorophyta).

To understand the evolutionary driving forces of chloroplast (or plastid) genomes (plastomes) in the green macroalgal genus Ulva (Ulvophyceae, Chlorophyta), in this study, we sequenced and constructed seven complete chloroplast genomes from five Ulva species, and conducted comparative genomic analysis of Ulva plastomes in Ulvophyceae. Ulva plastome evolution reflects the strong selection pressure driving the compactness of genome organization and the decrease of overall GC composition. The overall plastome sequences including canonical genes, introns, derived foreign sequences and non-coding regions show a synergetic decrease in GC content at varying degrees. Fast degeneration of plastome sequences including non-core genes (minD and trnR3), derived foreign sequences, and noncoding spacer regions was accompanied by the marked decrease of their GC composition. Plastome introns preferentially resided in conserved housekeeping genes with high GC content and long length, as might be related to high GC content of target site sequences recognized by intron-encoded proteins (IEPs), and to more target sites contained by long GC-rich genes. Many foreign DNA sequences integrated into different intergenic regions contain some homologous specific orfs with high similarity, indicating that they could have been derived from the same origin. The invasion of foreign sequences seems to be an important driving force for plastome rearrangement in these IR-lacking Ulva cpDNAs. Gene partitioning pattern has changed and distribution range of gene clusters has expanded after the loss of IR, indicating that genome rearrangement was more extensive and more frequent in Ulva plastomes, which was markedly different from that in IR-containing ulvophycean plastomes. These new insights greatly enhance our understanding of plastome evolution in ecologically important Ulva seaweeds.

Bacterial Contamination of Open Fractures: Pathogens and Antibiotic Resistance Patterns in East China.

Bacterial contamination of soft tissue in open fractures leads to high infection rates. Pathogens and their resistance against therapeutic agents change with time and vary in different regions. The purpose of this study was to characterize the bacterial spectrum present in open fractures and analyze the bacterial resistance to antibiotic agents based on five trauma centers in East China. A retrospective multicenter cohort study was conducted in six major trauma centers in East China from January 2015 to December 2017. Patients who sustained open fractures of the lower extremities were included. The data collected included the mechanism of injury, the Gustilo-Anderson classification, the isolated pathogens and their resistance against therapeutic agents, as well as the prophylactic antibiotics administered. In total, 1348 patients were included in our study, all of whom received antibiotic prophylaxis (cefotiam or cefuroxime) during the first debridement at the emergency room. Wound cultures were taken in 1187 patients (85.8%); the results showed that the positive rate of open fracture was 54.8% (651/1187), and 59% of the bacterial detections occurred in grade III fractures. Most pathogens (72.7%) were sensitive to prophylactic antibiotics, according to the EAST guideline. Quinolones and cotrimoxazole showed the lowest rates of resistance. The updated EAST guidelines for antibiotic prophylaxis in open fracture (2011) have been proven to be adequate for a large portion of patients, and we would like to suggest additional Gram-negative coverage for patients with grade II open fractures based on the results obtained in this setting in East China.

Biomimetic hydrogel derived from decellularized dermal matrix facilitates skin wounds healing.

Cutaneous wound healing affecting millions of people worldwide represents an unsolvable clinical issue that is frequently challenged by scar formation with dramatical pain, impaired mobility and disfigurement. Herein, we prepared a kind of light-sensitive decellularized dermal extracellular matrix-derived hydrogel with fast gelling performance, biomimetic porous microstructure and abundant bioactive functions. On account of its excellent cell biocompatibility, this ECM-derived hydrogel could induce a marked cellular infiltration and enhance the tube formation of HUVECs. In vivo experiments based upon excisional wound splinting model showed that the hydrogel prominently imparted skin wound healing, as evidenced by notably increased skin appendages and well-organized collagen expression, coupled with significantly enhanced angiogenesis. Moreover, the skin regeneration mediated by this bioactive hydrogel was promoted by an accelerated M1-to-M2 macrophage phenotype transition. Consequently, the decellularized dermal matrix-derived bioactive hydrogel orchestrates the entire skin healing microenvironment to promote wound healing and will be of high value in treatment of cutaneous wound healing. As such, this biomimetic ddECMMA hydrogel provides a promising versatile opinion for the clinical translation.

Tandem integration of circular plasmid contributes significantly to the expanded mitochondrial genomes of the green-tide forming alga Ulva meridionalis (Ulvophyceae, Chlorophyta).

Comparative mitogenomics of Ulva species have revealed remarkable variations in genome size due to the integration of exogenous DNA fragments, the proliferation of group I/II introns, and the change of repeat sequences. The genus Ulva is a species-rich taxonomic group, containing a variety of green-tide forming algae. In this study, five complete mitogenomes of the green-tide forming macroalga, Ulva meridionalis R. Horimoto and S. Shimada, were assembled and compared with the available ulvophyceae mtDNAs. The main circular mitogenomes of U. meridionalis ranged from 82.94 to 111.49 kb in size, and its 111.49-kb mitogenome was the largest Ulva mitogenome sequenced so far. The expansion of U. meridionalis mitogenomes is mainly due to the tandem integration of a 5.36-kb mitochondrial circular plasmid (pUme), as well as the proliferation of introns. An intact DNA-directed RNA polymerase gene (rpo) was present in pUme of U. meridionalis and was then detected in two putative plasmids (pUmu1 and pUmu2) found in Ulva mutabilis. The observed integration of the circular plasmid into U. meridionalis mitogenomes seems to occur via homologous recombination, and is a more recent evolutionary event. Many highly homologous sequences of these three putative plasmids can be detected in the other Ulva mtDNAs sequenced thus far, indicating the integration of different mitochondrial plasmid DNA into the mitogenomes is a common phenomenon in the evolution of Ulva mitogenomes. The random incidence of destruction of plasmid-derived rpos and open reading frames (orfs) suggests that their existence is not the original characteristic of Ulva mitogenomes and there is no selective pressure to maintain their integrity. The frequent integration and rapid divergence of plasmid-derived sequences is one of the most important evolutionary forces to shape the diversity of Ulva mitogenomes.

Simultaneous reconstruction of septic composite defects in lower extremities: Combination of fasciocutaneous perforator flap and Masquelet technique.

Background: Management of composite defects with deep infection is a challenge to reconstructive surgeons. This study aimed to demonstrate the versatility, safety, and complications of simultaneous reconstruction of infectious composite defects with fasciocutaneous perforator flap combined with the Masquelet technique. Methods: This study presents 10 patients in whom a fasciocutaneous perforator flap combined with the Masquelet technique was used to restore soft tissue and bone defects of the lower extremity, and were admitted in two level 1 trauma centers in Shanghai. The first stage included debridement of necrotic bone and infected tissues, implantation of a polymethylmethacrylate cement spacer to cover the void; bridging fixation of the osseous defect using external or internal fixators, and soft-tissue reconstruction with a fasciocutaneous perforator flap. The second stage included cement spacer removal with membrane preservation, refreshing bone edges, and grafting the cavity with bone morphogenetic proteins and autologous iliac bone graft. Results: The mean follow-up duration after autologous bone graft was 17.5 months. The average bony defects and average flap dimensions were 7.1 cm and 44.9 cm2, respectively. All flaps survived uneventfully. No recurrence of infection was detected in either the second stage of surgery or follow-up period. The mean duration of bone consolidation was 31.9 weeks. One patient had a 2 cm leg length discrepancy, and one patient had mild foot drop. No residual deformity requiring a secondary procedure occurred. Conclusion: Fasciocutaneous perforator flap combined with Masquelet technique provides a reliable and versatile alternative for patients with composite defects resulting from lower extremity infection.

miR-199a-5p Plays a Pivotal Role on Wound Healing via Suppressing VEGFA and ROCK1 in Diabetic Ulcer Foot.

Diabetes mellitus (DM) is a growing health problem. As a common complication of DM, diabetic foot ulcer (DFU) results in delayed wound healing and is a leading cause of nontraumatic amputation. miR-199a-5p, a short noncoding RNA, had abnormal expression in DFU wound tissues. The expression of miR-199a-5p was significantly increased in DFU wound tissues, skin tissues of diabetic rats, and high glucose-induced cells. Vascular endothelial growth factor A (VEGFA) and Rho-associated kinase 1 (ROCK1) are directly targets of miR-199a-5p. Inhibiting the expression of miR-199a-5p alleviated the inhibition of VEGFA and ROCK1, thereby rescued impaired proliferation and migration of HG-induced cells, and restored the normal function of the cells to some extent. In diabetic rats, inhibition of miR-199a-5p significantly increased the expression of VEGFA and ROCK1, significantly promoted wound healing, and rescued impaired wound healing. miR-199a-5p and its targets showed therapeutic effect on diabetic wounds.

N klA Case Report on a IV-degree Thermal Crush Injury of Right Upper Arm: The Application of Functional Prosthesis Implantation Technology.

Severe IV-degree thermal crush injury of limbs involved the subcutaneous fascia, muscle and bone, which may lead to amputation and has a great impact on the patient's quality of life. We can repair wounds with pedicle flaps or even free flaps, However, there are still huge challenges in bone defect of extremities and functional reconstruction. In recent years, with the development of functional prostheses, we have reconstructed limb functions in many patients helping them to complete their daily lives. We report a case where the right upper arm was injured by thermal crush, leading severe burns to the skin, fascia, muscle and bone. We applied a pedicled latissimus dorsi flap and a free anterolateral thigh flap to repair the wound, and realized the function of limb salvage and movement of the right upper arm by implanting 3D printed scapula, upper arm, and elbow joint prostheses. This case illustrates that IV-degree burns involving bones have new technologies to repair and achieve mobility now.

Phylogenomic analysis of the chloroplast genome of the green-tide forming macroalga Ulva intestinalis Linnaeus (Ulvophyceae, Chlorophyta).

Ulva intestinalis Linnaeus 1753 (Ulvophyceae, Chlorophyta) is a marine green macroalga that is distributed on coasts of the Yellow Sea and the Bohai Sea in China. Here, the complete chloroplast genome of U. intestinalis was constructed and analyzed comparatively. The chloroplast genome of U. intestinalis is a 99,041-bp circular molecule that harbors a total of 112 genes including 71 protein-coding genes (PCGs), 26 transfer RNA genes (tRNAs), three ribosomal RNA genes (rRNAs), three free-standing open reading frames (orfs) and nine intronic orfs, and ten introns in seven genes (atpA, infA, psbB, psbC, petB, rrnL, and rrnS). The maximum likelihood (ML) phylogenomic analysis shows that U. intestinalis firstly groups with Ulva compressa, and then these two species together with the Ulva australis-Ulva fenestrata-Ulva rotundata subclade form a monophyletic clade, Ulva lineage II. U. intestinalis chloroplast genome is the only one in Ulva lineage II where the reversal of a collinear block of two genes (psbD-psbC) did not occur, and its genome structure is consistent with that of most chloroplast genomes in Ulva lineage I, indicating that the similarity of genome structure is not completely related to the genetic relationship of Ulva species. Our genomic data will facilitate the development of specific high-resolution chloroplast molecular markers for rapid identification of U. intestinalis, and help us understand its population diversity and genetic characteristics on a global scale.

miR-224-5p-enriched exosomes promote tumorigenesis by directly targeting androgen receptor in non-small cell lung cancer.

Non-small cell lung cancer (NSCLC) is the most common form of cancer, resulting in cancer-related deaths worldwide. Exosomes, a subclass of extracellular vesicles, are produced and secreted from various types of cells, including cancer cells. Cancer-derived exosomes can deliver nucleic acids, proteins, and lipids to provide a favorable microenvironment that supports tumor growth through enhancing cell proliferation and metastasis. Our results showed that miR-224-5p was upregulated in NSCLC patient tissues and cell lines, with a tumor-promoting phenotype. Meanwhile, exosome-derived miR-224-5p induced cell proliferation and metastasis in NSCLC and human lung cells. Moreover, we characterized the androgen receptor (AR) as a direct target of miR-224-5p. Tumor xenograft assay experiments revealed that overexpression of miR-224-5p drove NSCLC tumor growth via the suppression of AR and the mediation of epithelial-mesenchymal transition (EMT). Collectively, our results suggest that miR-224-5p-enriched exosomes promote tumorigenesis by directly targeting AR in NSCLC, which may provide novel potential therapeutic and preventive targets for NSCLC.

Locking Plate Use with or without Strut Support for Varus Displaced Proximal Humeral Fractures in Elderly Patients.

Varus displaced fractures of the proximal part of the humerus, particularly in osteoporotic bone, commonly require open reduction and internal fixation. However, surgical treatment methods remain controversial and have shown inconsistent results. A fibular allograft for indirect medial reduction and strut support has been used in an effort to prevent secondary postoperative varus displacement. However, the long-term outcomes of this method require confirmation. We hypothesized that placing a fibular strut parallel to the calcar screw could increase the biomechanical stability of the medial hinge, thus preventing secondary varus deformity. In the present study, we compared the clinical outcomes of locking plate use with and without medial strut support with use of a fibular allograft for the treatment of varus humeral fractures in patients ≥65 years old. METHODS: We compared 2 different graft techniques involving the use of fibular allografts in elderly patients with varus displaced proximal humeral fractures who underwent open reduction and internal fixation. The patients were divided into 3 groups: (1) the intramedullary graft group (Group A), (2) the medial hinge support group (Group B), and (3) the locking plate alone group (Group C). Clinical outcomes included the final varus angulation of the humeral head, the occurrence of major complications (screw cut-out or cut-through or osteonecrosis), and the American Shoulder and Elbow Surgeons (ASES) score at 1 year after treatment. RESULTS: A total of 128 patients were included in our study. The final varus angles were 14.7°, 13.1°, and 18.6°, for the intramedullary graft group, the medial hinge support group, and locking plate alone group, respectively. The mean ASES scores were 87.2, 88.6, and 82.2, respectively. There were differences in ASES scores between Group A and Group C as well as also between Group B and Group C. Fewer major complications were found in patients managed with locking plates in combination with intramedullary graft or medial hinge support (Group A and Group B) than in patients managed with locking plates alone (Group C). CONCLUSIONS: The use of a locking plate in combination with medial strut support with use of a fibular allograft reduced complications when used for the treatment of varus displaced proximal humeral fractures in elderly patients in comparison with the use of a locking plate alone. LEVEL OF EVIDENCE: Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.

Establishing an Evaluation System and Limb-Salvage Protocol for Mangled Lower Extremities in China.

Road traffic accident-related severely injured extremities account for the majority of disabilities in young people in China. Limb-salvage concepts and techniques vary greatly from physician to physician and from district to district in China. Current severity-scoring systems for lower-extremity injuries lack sensitivity and cannot be used as the sole criterion by which amputation decisions are made. China lacks a national database of mangled lower extremities, which is a priority for both limb-salvage protocols and scoring system development.

Study of Recellularized Human Acellular Arterial Matrix Repairs Porcine Biliary Segmental Defects.

Background: With the popularity of laparoscopic cholecystectomy, common bile duct injury has been reported more frequently. There is no perfect method for repairing porcine biliary segmental defects. Methods: After the decellularization of human arterial blood vessels, the cells were cultured with GFP+ (carry green fluorescent protein) porcine bile duct epithelial cells. The growth and proliferation of porcine bile duct epithelial cells on the human acellular arterial matrix (HAAM) were observed by hematoxylin-eosin (HE) staining, electron microscopy, and immunofluorescence. Then, the recellularized human acellular arterial matrix (RHAAM) was used to repair biliary segmental defects in the pig. The feasibility of it was detected by magnetic resonance cholangiopancreatography, liver function and blood routine changes, HE staining, immunofluorescence, real-time quantitative PCR (RT-qPCR), and western blot. Results: After 4 weeks (w) of co-culture of HAAM and GFP+ porcine bile duct epithelial cells, GFP+ porcine bile duct epithelial cells grew stably, proliferated, and fused on HAAM. Bile was successfully drained into the duodenum without bile leakage or biliary obstruction. Immunofluorescence detection showed that GFP-positive bile duct cells could still be detected after GFP-containing bile duct cells were implanted into the acellular arterial matrix for 8 w. The implanted bile duct cells can successfully resist bile invasion and protect the acellular arterial matrix until the newborn bile duct is formed. Conclusion: The RHAAM can be used to repair biliary segmental defects in pigs, which provides a new idea for the clinical treatment of common bile duct injury.