Amino acid analysis as a method of discovering biomarkers for diagnosis of diabetes and its complications.

Diabetes mellitus (DM) is a severe chronic diseases with a global prevalence of 9%, leading to poor health and high health care costs, and is a direct cause of millions of deaths each year. The rising epidemic of diabetes and its complications, such as retinal and peripheral nerve disease, is a huge burden globally. A better understanding of the molecular pathways involved in the development and progression of diabetes and its complications can facilitate individualized prevention and treatment. High diabetes mellitus incidence rate is caused mainly by lack of non-invasive and reliable methods for early diagnosis, such as plasma biomarkers. The incidence of diabetes and its complications in the world still grows so it is crucial to develop a new, faster, high specificity and more sensitive diagnostic technologies. With the advancement of analytical techniques, metabolomics can identify and quantify multiple biomarkers simultaneously in a high-throughput manner, and effective biomarkers can greatly improve the efficiency of diabetes and its complications. By providing information on potential metabolic pathways, metabolomics can further define the mechanisms underlying the progression of diabetes and its complications, help identify potential therapeutic targets, and improve the prevention and management of T2D and its complications. The application of amino acid metabolomics in epidemiological studies has identified new biomarkers of diabetes mellitus (DM) and its complications, such as branched-chain amino acids, phenylalanine and arginine metabolites. This study focused on the analysis of metabolic amino acid profiling as a method for identifying biomarkers for the detection and screening of diabetes and its complications. The results presented are all from recent studies, and in all cases analyzed, there were significant changes in the amino acid profile of patients in the experimental group compared to the control group. This study demonstrates the potential of amino acid profiles as a detection method for diabetes and its complications.

Myocardin suppression increases lipid retention and atherosclerosis via downregulation of ABCA1 in vascular smooth muscle cells.

Myocardin (MYOCD) plays an important role in cardiovascular disease. However, its underlying impact on atherosclerosis remains to be elucidated. ATP binding cassette transporter A1 (ABCA1), a key membrane-associated lipid transporter which maintains intracellular lipid homeostasis, has a protective function in atherosclerosis progress. The purpose of this study was to investigate whether and how the effect of MYOCD on atherosclerosis is associated with ABCA1 in vascular smooth muscle cells (VSMCs). We found both MYOCD and ABCA1 expression were dramatically decreased in atherosclerotic patient aortas compared to control. MYOCD knockdown inhibited ABCA1 expression in human aortic vascular smooth muscle cells (HAVSMCs), leading to reduced cholesterol efflux and increased intracellular cholesterol contents. MYOCD overexpression exerted the opposite effect. Mechanistically, MYOCD regulates ABCA1 expression in an SRF-dependent manner. Consistently, apolipoprotein E-deficient mice treated with MYOCD shRNA developed more plaques in the aortic sinus, which is associated with reduced ABCA1 expression, increased cholesterol retention in the aorta, and decreased high-density lipoprotein cholesterol levels in the plasma. Our data suggest that MYOCD deficiency exacerbates atherosclerosis by downregulating ABCA1 dependent cholesterol efflux from VSMCs, thereby providing a novel strategy for the therapeutic treatment of atherosclerotic cardiovascular disease.

Knockdown of miR-660 protects nucleus pulposus cells from TNF-a-induced apoptosis by targeting serum amyloid A1.

BACKGROUND: Intervertebral disc degeneration (IVDD) is a well-known cause of lower back pain, which is induced by multiple factors including increased apoptosis and decreased survival of nucleus pulposus cells. In this study, we evaluate the effect and potential mechanism of miR-660 on the nucleus pulposus cells apoptosis induced by TNF-α. METHODS: First, we collected tissue of nucleus pulposus from IVDD and healthy controls. General characteristic of the IVDD and healthy control was also collected. And, we also collected nucleus pulposus cells that stimulated by TNF-α or control. miRNA microarray was performed to identify the differentially expressed miRNAs. Apoptosis rate and miR-660 relative expression was measured after stimulated with different concentration of TNF-α to identify the optimal concentration of TNF-α. Second, we successfully constructed antigomiR-660 to block the miR-660 expression in nucleus pulposus cells and then stimulated with TNF-α (100 ng/ml, 12 h). The apoptosis rates and relative protein expression were then measured again. The target association between miR-660 and SAA1 was confirmed by dual-luciferase reporter. RESULTS: There was no significant difference between the age (IVDD: 39 ± 10 years, healthy controls: 36 ± 7 years), BMI and sex between IVDD and healthy controls. Microarray analysis found that miR-660 was significantly up-regulated in IVDD and TNF-α treated groups, which was further identified by PCR. We found that the rate of apoptosis and miR-660 expression increased with TNF-α concentration increased. Finally, TNF-a with 100 ng/ml was used for further experiment. Compared with TNF-α group, TNF-α + antigomiR-660 could significantly down-regulated the apoptosis rate and relative protein (c-Caspase3 and c-Caspase7). Dual-luciferase reporter revealed that miR-660 could directly binding to the SAA1 at 80-87 sites. Compared with TNF-α alone group, TNF-α + antigomiR-660 significantly up-regulated the SAA1 expression (P < 0.05). CONCLUSION: These results indicated that knockdown of miR-660 protected the nucleus pulposus from apoptosis that induced TNF-α via up-regulation of SAA1. Further studies should focus on the role of miR-660 in protecting IVDD in vivo.

Tripaddle Posterior Interosseous Artery Flap Design for 3-Finger Defects: An Evaluation of 3 Surgical Approaches.

INTRODUCTION: The tripaddle posterior interosseous artery (PIA) flap can be used for multifinger defect resurfacing, but interpatient variations in perforator distribution remain an ongoing challenge when using this approach. This study aims to evaluate the efficacy of 3 different tripaddle PIA perforator flap designs according to the PIA perforator distribution for the repair of 3-finger defects. METHODS: In accordance with the size of the 3-finger defects and the position of the perforators, a tripaddle flap was designed on the multiple perforators of the descending branch of the PIA in the distal two thirds of the forearm. Patients received 1 of 3 distinct tripaddle PIA perforator flap designs based on perforator distributions of the PIA. RESULTS: Three cases of 3-finger defects were repaired with type A trefoil-shaped tripaddle flaps, whereas 4 cases were repaired with type B modified trefoil-shaped tripaddle flaps, and the other 3 cases were repaired with type C chain-shaped tripaddle flaps. All flaps survived except 2 paddles with tip necrosis. After 9.1 months of mean follow-up, 9 of the 10 cases demonstrated satisfactory cosmetic appearance, whereas the last case required a debulking procedure in the second stage. CONCLUSIONS: The free tripaddle PIA perforator flap is an effective option for repairing 3-finger skin defects. Various flap designs based on the PIA perforator distribution allow for more individualized treatment approaches.

Free multilobed posterior interosseous artery perforator flap for multi-finger skin defect reconstruction.

BACKGROUND: The posterior interosseous artery (PIA) perforator flap can be used for reconstruction of soft-tissue defects of fingers. Based on the multiple perforators from the posterior interosseous artery, we describe a technique to reconstruct the multi-finger defect in the use of the free multilobed PIA perforator flap. METHODS: PIA perforators from different areas of the forearm were used to design a free multilobed skin paddle for multi-finger skin defect reconstruction. Each paddle without the deep fascia had separate perforators. To increase the perforator pedicle length, the courses of the PIA perforators were dissected from the superficial layer of the deep fascia to the subcutaneous layer. RESULTS: The flap was raised as a unilateral free bilobed PIA perforator flap in 10 cases of two-finger defects, a free trilobed PIA perforator flap in two cases of three-finger defects, and a bilateral free bilobed PIA perforator flap in one case of four-finger defects. The average effective vascular pedicle length and trunk pedicle length were 8.3 and 3.1 cm, respectively, for the bilobed flap, and 6.3 and 4.0 cm, respectively, for the trilobed flap. All flaps survived except one paddle with tip necrosis. At 10.8 months (range, 4-27 months) after surgery, 10 cases showed satisfactory cosmetic appearance, while the fingers were bulky in the remaining three cases. The average score of static two-point discrimination in 10 innervated paddles was 12.9 mm. The remaining 20 paddles recovered only protective sensation. The average total active motion (TAM) of each finger was 164° before surgery and 187° at the latest follow-up. CONCLUSIONS: Free multilobed PIA perforator flap is a good candidate for reconstruction of multi-finger skin defect. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, Ⅳ.

[Microvascular anastomotic anterolateral thigh flaps for reconstruction of traumatic widespread defects of soft tissue in heel].

OBJECTIVE: To explore the results of repairing widespread defects of traumatic soft tissue in heel by microvascular anastomotic anterolateral thigh flaps. METHODS: Twenty-six consecutive free anterolateral thigh flaps in 26 patients were transplanted for repairing widespread defects of traumatic soft tissue in heel from October 1997 to March 2005, suturing the descending branch of the lateral circumflex femoral vascular and posterior tibial vascular to reconstruct the blood supply of transplanted flaps, repairing lateral femoral cutaneous nerve to recover their sensation, fixing tensor fasciae in the calcaneum to add their stabilization. RESULTS: All flaps survived completely,the wounds healed in the initial treatment, follow up 3 to 48 months, twenty-six cases achieved partial sensation, good contour and stabilization. CONCLUSIONS: The anterolateral thigh flap is an ideal flap for repairing widespread defects of traumatic soft tissue in heel, because it has such advantages as adequate blood supply, big dermatosis area and covert donor site, furthermore, nervi cutaneous femoris lateralis and tensor fasciae offer the good sensation and adequate stabilization.

[Transplantation of free latissimus dorsi myocutaneous flaps for craniomaxillofacial reconstruction].

OBJECTIVE: This clinical study was to improve the surgical treatment to craniomaxillofacial tissue defects. METHODS: Since 1997, eight cases with severe craniomaxillofacial defects were treated using free latissimus dorsi myocutaneous flaps. In the operation, nerve anastomosis was performed. Of the 8 cases, 7 were treated in one stage, 1 was treated in 3 steps. The craniomaxillofacial defects ranged from 10 cm x 8 cm to 30 cm x 12 cm. The flaps was 12 cm x 10 cm to 32 cm x 16 cm in size. RESULTS: Postoperative follow-up for 6 months to 4 years demonstrated satisfactory results in all the cases. There was neither necrosis nor ulcer after the operation. The sensation recovery of the flap was also satisfactory. CONCLUSION: Free transfer of the latissimus dorsi myocutaneous flap is an ideal treatment to severe craniomaxillofacial defects as it possesses the advantages of reliable blood supply, ability against infections, large size, concealed donor site, and functional restoration of sensation and movement.