Comprehensive metabolome and transcriptome analyses demonstrate divergent anthocyanin and carotenoid accumulation in fruits of wild and cultivated loquats.

Eriobotrya is an evergreen fruit tree native to South-West China and adjacent countries. There are more than 26 loquat species known in this genus, while E. japonica is the only species yet domesticated to produce fresh fruits from late spring to early summer. Fruits of cultivated loquat are usually orange colored, in contrast to the red color of fruits of wild E. henryi (EH). However, the mechanisms of fruit pigment formation during loquat evolution are yet to be elucidated. To understand these, targeted carotenoid and anthocyanin metabolomics as well as transcriptomics analyses were carried out in this study. The results showed that ß-carotene, violaxanthin palmitate and rubixanthin laurate, totally accounted for over 60% of the colored carotenoids, were the major carotenoids in peel of the orange colored 'Jiefangzhong' (JFZ) fruits. Total carotenoids content in JFZ is about 10 times to that of EH, and the expression levels of PSY, ZDS and ZEP in JFZ were 10.69 to 23.26 folds to that in EH at ripen stage. Cyanidin-3-O-galactoside and pelargonidin-3-O-galactoside were the predominant anthocyanins enriched in EH peel. On the contrary, both of them were almost undetectable in JFZ, and the transcript levels of F3H, F3'H, ANS, CHS and CHI in EH were 4.39 to 73.12 folds higher than that in JFZ during fruit pigmentation. In summary, abundant carotenoid deposition in JFZ peel is well correlated with the strong expression of PSY, ZDS and ZEP, while the accumulation of anthocyanin metabolites in EH peel is tightly associated with the notably upregulated expressions of F3H, F3'H, ANS, CHS and CHI. This study was the first to demonstrate the metabolic background of how fruit pigmentations evolved from wild to cultivated loquat species, and provided gene targets for further breeding of more colorful loquat fruits via manipulation of carotenoids and anthocyanin biosynthesis.

[Locking loop stitch with suture-bridge technique in repair of acute closed distal Achilles tendon rupture by using suture anchors].

OBJECTIVE: To explore clinical efficacy of Locking loop stitch with suture-bridge technique in repair of acute closed distal Achilles tendon rupture by using suture anchors. METHODS: From July 2019 to March 2021, 20 patients with acute closed distal Achilles tendon rupture were treated by minimally invasive suture anchor locking suture bridging repair technique. Among them, including 18 males and 2 females, aged from 19 to 52 years old with an average of(40.0±9.0) years old. Complications were observed, and recovery of ankle function was evaluated by American Orthopaedic Foot & Ankle Society(AOFAS) ankle and hindfoot function scoring system before operation and 1 year after operation. RESULTS: All patients followed up from 6 to 18 months with an average of (12.0±3.2) months. The incisions were healed at stageⅠwithout infection and skin necrosis occurred;no gastrocnemius nerve injury and deep vein thrombosis of the lower extremities occurred;and no heel pain and Achilles tendon re-rupture occurred. AOFAS scores of ankle and hindfoot increased from(59.0±4.3) before opertaion to(95.1±2.6) at 1 year after operation (t=-32.1, P<0.05). CONCLUSION: The effect of locking suture bridging with suture anchor nails to repair acute distal Achilles tendon rupture is definite, and it could reduce incidence of complications such as Achilles tendon re-rupture, nerve injury, and skin necrosis, which has advantages of small surgical trauma, reliable anastomosis method and good functional recovery, and is an ideal method for treating acute closed distal Achilles tendon rupture.

The protective effects of exenatide against AGEs-induced articular matrix degradation in human primary chondrocytes.

Osteoarthritis (OA) presents a major global health burden and is projected to become even more prevalent in coming decades. Therefore, it is of utmost importance to uncover novel therapies for the treatment and prevention of this disease. In the present study, we investigated the effects of exenatide, a specific glucagon-like peptide (GLP) agonist, on degradation of type II collagen and aggrecan, the two main components of the articular extracellular matrix, in human primary chondrocytes. Our results reveal that exenatide could ameliorate degradation of type II collagen and aggrecan by inhibiting expression of metalloproteinases (MMPs) and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) induced by advanced glycation end-products. We also found that exenatide reduces oxidative stress and inhibits activation of nuclear factor-κB through the p38 cellular signaling pathway. Taken together, the findings of this study indicate that exenatide may have potential as a novel treatment for osteoarthritis.

Resveratrol protects cardiomyocytes against anoxia/reoxygenation via dephosphorylation of VDAC1 by Akt-GSK3 ß pathway.

Our previous studies showed that the effect of resveratrol preventing mitochondrial permeability transition pore (mPTP) opening in myocardial ischemia/reperfusion injury was achieved by regulating voltage-dependent anion channel 1 (VDAC1). However, the underlying mechanism remains unclear. Previous studies demonstrated that the activity and function of VDAC1 are highly regulated by post-translational modification. In present study, we investigated whether resveratrol modulates VDAC1 phosphorylation to achieve cardioprotection and explored the signaling pathways involved. Our findings demonstrated that anoxia/reoxygenation (A/R) treatment, an ischemia/reperfusion model in vitro, enhanced VDAC1 phosphorylation in cardiomyocytes. Moreover, we found phosphorylated VDAC1 showed increased affinity to Bax, whereas interaction with hexokinase 2 (HK2) was reduced. Accordingly, the generation of reactive oxygen species increased, the mitochondrial membrane potential collapsed, mPTP opening increased and cytochrome c released into cytoplasm, thereby leading to increased apoptosis. Moreover, our data showed that pretreatment with resveratrol prior to A/R injury inhibited VDAC1 phosphorylation. Dephosphorylated VDAC1 using pretreated resveratrol promoted dissociation with Bax and binding to HK2, which subsequently protected cardiomyocytes against A/R injury. In addition, Akt and its downstream glycogen synthase kinase 3 ß (GSK3ß) were phosphorylated by the action of resveratrol. Akt inhibitor IV abrogated Akt-GSK3ß phosphorylation and thereby abolished the dephosphorylation activity of resveratrol on VDAC1. Moreover, all resveratrol-mediated protective effects on A/R injured cardiomyocytes were abolished by Akt inhibitor IV. Taken together, our data indicated that A/R injury enhanced VDAC1 phosphorylation in cardiomyocytes, whereas pretreatment with resveratrol dephosphorylated VDAC1 through the Akt-GSK3ß pathway, thereby protecting cardiomyocytes against A/R injury.

[Application of delayed skin-stretching device in treatment of skin and soft tissue defects].

Objective: To summarize the effectiveness of delayed skin-stretching device in treatment of skin and soft tissue defects. Methods: Between December 2014 and December 2016, 10 cases of skin and soft tissue defects were treated with delayed skin-stretching device. There were 6 males and 4 females with an average age of 53 years (range, 42-64 years). The skin and soft tissue defects were caused by acute trauma in 6 cases. The incision could not be closed directly after making incisions because of osseous fascia syndrome in 3 cases. The skin soft tissue defect caused after huge carbuncle incision and drainage in 1 case. The defect located at thigh in 4 cases, lower leg in 3 cases, upper arm in 2 cases, back in 1 case. The defect area ranged from 10 cm×4 cm to 22 cm×12 cm. Pinch test was performed on the wound margin, which confirmed that the wound could not be closed directly. Results: Tension blisters were found in 3 cases during traction, and no complications such as impaired blood circulation or skin necrosis occurred in all cases. Skin defects closed directly after continuously stretching for 7-18 days. No skin graft or free flap repair was performed in all patients. The wound healed well after operation. All the 10 patients were followed up 5-8 months (mean, 6.5 months). There was no necrosis around the wound margin and the scar was linear. The sensation and function were not affected. Conclusion: Delayed skin-stretching device is an effective method to treat skin and soft tissue defects, which has the advantages of simple operation, lower risk of operation, less complications, and reliable effectiveness.

VDAC1 deacetylation is involved in the protective effects of resveratrol against mitochondria-mediated apoptosis in cardiomyocytes subjected to anoxia/reoxygenation injury.

We have recently demonstrated that Voltage-dependent anion channel 1 (VDAC1), a protein located in the mitochondrial outer membrane, is involved in the effects of resveratrol on the mitochondrial permeability transition pore (mPTP). However, the underlying mechanism of action remains to be elucidated. In the present study, we demonstrated that resveratrol promoted VDAC1 deacetylation in cardiomyocytes in response to anoxia/reoxygenation (A/R) injury. Moreover, silent information regulator of transcription 1 (SIRT1), a NAD+-dependent class III histone deacetylase, was up-regulated after pretreatment with resveratrol. Cells that were treated with Ex527, a specific inhibitor of SIRT1, showed a reduction in both SIRT1 expression and VDAC1 deacetylation, indicating that the deacetylation effect of resveratrol on VDAC1 is mediated by SIRT1. Furthermore, the ability deacetylated VDAC1 to bind to Bax was decreased after pretreatment with resveratrol, whereas Bcl-2 expression changed in the opposite direction. As a result, opening of the mPTP was restrained, the mitochondrial membrane potential was reserved, and cytochrome c release was inhibited, which subsequently decreased cardiomyocyte apoptosis. However, the cardioprotective effects observed after treatment of resveratrol could be abrogated by Ex527. In conclusion, resveratrol induces deacetylation of VDAC1 by SIRT1, thereby preventing mitochondria-mediated apoptosis in cardiomyocytes upon A/R injury.

Soybean isoflavone treatment induces osteoblast differentiation and proliferation by regulating analysis of Wnt/ß-catenin pathway.

It has been reported that soybean isoflavones (SI) have anti-bone resorptive activity in vivo. However, little is known about the cellular and molecular mechanisms of this effect. In this study, we investigated the effects of SI on osteoblast differentiation and proliferation. Results showed that SI promoted osteoblast proliferation and differentiation which could be marked as the formation of hydroxyapatite crystals or extracellular matrix and increased ALP level. SI treatment markedly decreased RANKL levels and increased OPG levels in osteoblast cells. SI resulted in activation of the Wnt/ß-catenin pathway. These results demonstrate that SI may be a useful remedy for the treatment of osteoporosis.

Chemical characterization and varying level of the proteoglycans.

Proteoglycans are a major component of the animal extracellular matrix, the "filler" substance existing between cells in an organism. Here they form large complexes, both to other proteoglycans, to hyaluronan and to fibrous matrix proteins (such as collagen). Proteoglycans are heavily glycosylated glycoproteins. In this study, we evaluate effect of collagen and proteoglycan on osteoporosis. Rats were treated with ovariectomy (OVX) for 4 weeks to induce osteoporosis. In OVX rats, the levels of collagen and proteoglycan in knee joint cartilage and anterior cruciate ligament were significantly decreased in the OVX group animals compared to the sham group. OVX rats showed decreased levels of magnesium (Mg(2+)) and calcium (Ca(2+)), increased concentrations of alkaline phosphatase (ALP) and phosphorus (P) in the serum. We conclude that osteoporosis is closely associated to decreased collagen and proteoglycan levels in knee joint cartilage and anterior cruciate ligament.

Effect of ischemia post-conditioning on skeletal muscle oxidative injury, mTOR, Bax, Bcl-2 proteins expression, and HIF-1α/ß-actin mRNA, IL-6/ß-actin mRNA and caveolin-3/ß-actin mRNA expression in ischemia-reperfusion rabbits.

This study is designed to investigate the effect of ischemia post-conditioning on IR-induced skeletal muscle injury in limbs of experimental rabbits. Rabbits are randomized to one of the following three groups: sham control, ischemic reperfusion, ischemic postconditioning. The lipid peroxidation level, antioxidant enzymes activities, skeletal muscle mammalian target of rapamycin (mTOR), Bax, Bcl-2 proteins expression and Bcl-2/Bax, and HIF-1α/ß-actin mRNA, interleukin-6 (IL-6)/ß-actin mRNA and caveolin-3/ß-actin mRNA expression were tested in the current study. The results suggested that ischemic postconditioning might decrease lipid peroxidation level, lactic dehydrogenase (LDH), creatine kinase (CK) activities, Bcl-2 proteins expression and Bcl-2/Bax, HIF-1α/ß-actin mRNA expression and increase skeletal muscle antioxidant enzymes activities, Bax protein expression and IL-6/ß-actin mRNA and caveolin-3/ß-actin mRNA expression. These indicated that protective effect of ischemic postconditioning against IR-induced skeletal muscle injury involve into a complex molecular mechanism. Our research results may offer a theoretical guidance for therapy of related clinical diseases.

Cistanches Herba aqueous extract affecting serum BGP and TRAP and bone marrow Smad1 mRNA, Smad5 mRNA, TGF-ß1 mRNA and TIEG1 mRNA expression levels in osteoporosis disease.

We studied molecular mechanism of Cistanches Herba aqueous extract (CHAE) in ovariectomized (OVX) rats, as an experimental model of postmenopausal osteoporosis. Female rats were either sham-operated or bilaterally OVX; and at 60 days postoperatively. The OVX group (n = 8) received an ovariectomy and treatment with normal saline for 90 days commencing from 20th post ovariectomy day. The ovariectomized +CHAE (OVX + CHAE) group (n = 8) received an ovariectomy and were treated with Cistanches Herba aqueous extract of 100 mg/kg body weight daily for 90 days commencing from 22nd post ovariectomy day. The ovariectomy +CHAE (OVX + CHAE) group (n = 8) received an ovariectomy, and were treated with the of 200 mg/kg body weight daily for 90 days commencing from 20th post ovariectomy day. Serum BGP and TRAP, E2, FSH and LH level, bone marrow Smad1, Smad5, TGF-ß1 and TIEG1 mRNA expression levels were examined. Results showed that serum BGP and TRAP, FSH and LH levels were significantly increased, whereas E2, Smad1, Smad5, TGF-ß1 and TIEG1 mRNA and proteins expression levels were significantly decreased in OVX rats compared to sham rats. 90 days of CHAE treatment could significantly decrease serum BGP and TRAP, FSH and LH levels, and increase E2, Smad1, Smad5, TGF-ß1 and TIEG1 mRNA and proteins expression levels in OVX rats. It can be concluded that CHAE play its protective effect against OVX-induced bone degeneration partly by regulating some bone metabolism related genes, e.g. Smad1, Smad5, TGF-ß1 and TIEG1.

Influence of ShuJinHuoXue tablets on ischemia reperfusion injury of animals' skeletal muscle.

Ischemia-reperfusion (IR) can lead to serious tissue oxidative injury in animals. ShuJinHuoXue tablet (SJHXT) is a Chinese Traditional Medicine which can relax the muscles and stimulate the blood circulation and has been used as a clinical medicine. In the present study, we investigated the effects of SJHXT pretreatment on oxidative injury using an animal model of acute limb IR. Results showed that SJHXT pre-treatment (200, 300 and 400 mg/kg/day) markedly reduced serum endothelin-1 (ET-1), thromboxane B2 (TXB2) levels and thromboxane B2/6-keto- prostaglandin F1α (TXB2/6-Keto-PGF(1α)), wet weight/dried weight (W/D) ratio, myeloperoxidase (MPO), creatine kinase (CK), lactate dehydrogenase (LDH) activities, and increased serum nitric oxide (NO), 6-Keto-PGF(1α) levels and NO/ET-1 ratio in the IR+SJHXT groups. In addition, the SJHXT pre-treatment (200, 300 and 400 mg/kg/day) markedly reduced skeletal muscle Ca²âº, malondialdehyde (MDA) levels, increased Na⁺-K⁺-ATPase, Ca²âº-Mg²âº-ATPase, superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities. Our results suggest that SJHXT pre-treatment may improve skeletal muscle blood vessel microcirculation, decrease skeletal muscle oxidative injury and enhance antioxidant enzymes activities in IR animals.

Lycopene effects on serum mineral elements and bone strength in rats.

This study investigated the beneficial effect of lycopene on bone biomarkers in ovariectomized (OVX) rats. Female Wistar rats were either sham operated or surgically ovariectomized and then fed with lycopene for 8 weeks. Serum Ca, P, alkaline phosphatase (ALP), interleukin 6 (IL-6) and bone gla protein (BGP) concentration was significantly higher in the untreated OVX group compared with that of the sham group, whereas serum estrogen levels were lower. Bone mineral density (BMD), BMD/wt, bone mineral content (BMC), BMC/wt values, maximum load, stiffness, energy and maximum stress were significantly lower in the untreated OVX group compared with that of the sham group. Administration of lycopene (20, 30 and 40 mg/kg b.w.) for 8 weeks significantly decreased serum Ca, P, ALP, and IL-6 concentration, and enhanced serum estrogen level, BMD, BMD/wt, BMC, BMC/wt values, maximum load, stiffness, energy and maximum stress in lycopene-treated OVX groups. In conclusion, the consumption of lycopene may have the most protective effect on bone in OVX rats.

Effect of Cistanches Herba aqueous extract on bone loss in ovariectomized rat.

To assess the ability of traditional Chinese medicine Cistanches Herba extract (CHE) to prevent bone loss in the ovariectomized (OVX) rat, Cistanches Herba extract (CHE) was administered intragastrically to the rats. Female rats were anesthetized with pentobarbital sodium (40 mg kg(-1), i.p.), and their ovaries were removed bilaterally. The rats in the sham-operated group were anesthetized, laparotomized, and sutured without removing their ovaries. After 1 week of recovery from surgery, the OVX rats were randomly divided into three groups and orally treated with H(2)O (OVX group) or CHE (100 or 200 mg kg(-1) daily) for 3 months. The sham-operated group (n = 8) was orally treated with H(2)O. After 3 months, the total body bone mineral density (BMD), bone mineral content (BMC), Bone biomechanical index, blood mineral levels and blood antioxidant enzymes activities were examined in sham-operated, ovariectomized and Cistanches Herba extract treated rats. Results showed that Cistanches Herba extract treatment significantly dose-dependently enhanced bone mineral density (BMD), bone mineral content (BMC), maximum load, displacement at maximum load, stress at maximum load, load at auto break, displacement at auto break, and stress at auto break, and blood antioxidant enzymes activities, decreased blood Ca, Zn and Cu levels compared to the OVX group. This experiment demonstrates that the administration of Cistanches Herba extract to ovariectomized rats reverses bone loss and prevents osteoporosis.

Salvage of amputated upper extremities with temporary ectopic implantation followed by replantation at a second stage.

Salvage of the complex amputation of extremities, such as combined with devastating segmental injuries, extensive soft tissue defect, and multiple important organ injuries, continues to be a challenge for plastic surgeons. Temporary ectopic implantation of the amputated part to a healthy recipient site allows the patient to recover from critical combined injuries, radical debridements, and soft tissue repair. In this article, the authors report two cases of temporary ectopic implantation of complexly amputated forearms, followed by successful replantation to their anatomic positions at a second stage. The contralateral upper extremity is an acceptable recipient site for temporary ectopic implantation. In secondary replantation, a cross-arm flap can be designed to carry the vascular pedicle from the ectopic implantation recipient to improve blood supply to the replanted part when the second blood supply is established. The authors validated that temporary ectopic implantation of amputated parts provides an alternative procedure for the salvage of amputated extremities under special circumstances.

[Application of the dorsoulnar arterial retrograde flap of the thumb to repair of soft tissue defect at the distal thumb].

OBJECTIVE: To introduce the surgical procedure and indication of the dorsoulnar arterial retrograde flap of the thumb in repair of soft tissue defect. METHODS: From March 2000 to March 2001, 12 cases of soft tissue defect with exposed bone at the distal thumb were involved. Ten flaps were pedicled distally at the proximal rotation point, 2.5 cm proximal to the cuticule. Two flaps were pedicled distally at the distal rotation point, 1.0 cm proximal to the cuticule. The flaps harvested in this study were 1.1 cm x 1.3 cm to 1.8 cm x 2.5 cm. RESULTS: All the flaps survived. After an follow-up of 2 to 14 months postoperatively, the appearance of the thumbs were satisfactory, except those of 2 thumbs repaired by the flaps transferred at the distal rotation point, which were bulkiness because of the uncovered pedicle. In 6 cases, the defect of finger pulp was repaired, and the 2-point discrimination was measured 8-10 mm. The same range of motion of the thumb IP joint were observed in both sides in all cases. CONCLUSION: The optimal indication of the procedure is soft tissue defect at the distal thumb.

[Temporary ectopic implantation of amputated foot].

OBJECTIVE: To investigate an effective technique of temporary ectopic implantation for amputated extremity under complex condition. METHODS: Two cases of amputated foot, which could not be implanted primarily, were treated with temporary ectopic implantation. The other leg of patient was chosen as recipient site. The posterotibial artery and saphenous vein were chosen as recipient vessels. When the general condition and the proximal condition of the amputated part were suitable, the ectopic implanted feet were transferred to their anatomic positions. RESULTS: All the feet survived after the replantation. The injured limbs recovered their normal length and sensation. The patients could walk after 4-6 months. CONCLUSION: Temporary ectopic implantation is an ideal technique for the salvage of amputated limb and organ under special condition. Severed foot and lower segment of the leg under complex condition were the best indication for the temporary ectopic implantation.