Eugenol Possesses Colitis Protective Effects: Impacts on the TLR4/MyD88/NF-[Formula: see text]B Pathway, Intestinal Epithelial Barrier, and Macrophage Polarization.

Eugenol (EU) has been shown to ameliorate experimental colitis due to its anti-oxidant and anti-inflammatory bioactivities. In this study, DSS-induced acute colitis was established and applied to clarify the regulation efficacy of EU on intestinal barrier impairment and macrophage polarization imbalance along with the inflammatory response. Besides, the adjusting effect of EU on macrophages was further investigated in vitro. The results confirmed that EU intervention alleviated DSS-induced colitis through methods such as restraining weight loss and colonic shortening and decreasing DAI scores. Microscopic observation manifested that EU maintained the intestinal barrier integrity in line with the mucus barrier and tight junction protection. Furthermore, EU intervention significantly suppressed the activation of TLR4/MyD88/NF-[Formula: see text]B signaling pathways and pro-inflammatory cytokines gene expressions, while enhancing the expressions of anti-inflammatory cytokines. Simultaneously, WB and FCM analyses of the CD86 and CD206 showed that EU could regulate the DSS-induced macrophage polarization imbalance. Overall, our data further elucidated the mechanism of EU's defensive effect on experimental colitis, which is relevant to the protective efficacy of intestinal barriers, inhibition of oxidative stress and excessive inflammatory response, and reprogramming of macrophage polarization. Hence, this study may facilitate a better understanding of the protective action of the EU against UC.

[Effect and mechanism of acupoint injection on influenza A virus induced pneumonia in mice].

OBJECTIVE: To investigate the effect and mechanism of acupoint injection with 0.1% vitamin C+vitamin B complex solution (VC+VBCo) at "Tiantu" (CV 22), "Quchi" (LI 11) and "Zusanli" (ST 36) in mouse model of pneumonia induced by influenza A virus (A/PR/8/34 [H1N1], PR8). METHODS: Sixty male ICR mice were randomized into 6 groups, i.e. control group, model group, acupoint injection group, intraperitoneal injection group, non-target point group and ribavirin group, 10 mice in each one. Except the control group, the pneumonia models were induced by slow nasal dripping PR8 virus in the other groups. On the 2nd day of experiment, VC+VBCo solution, 40 µL was injected at "Tiantu" (CV 22), "Quchi" (LI 11, left) and "Zusanli" (ST 36, left) in the acupoint injection group; VC+VBCo solution, 120 µL was injected intraperitoneally in the intraperitoneal injection group; VC+VBCo solution, 40 µL was injected at non-target acupoints (0.5 cm away from "Tiantu" [CV 22] to the left side, "Quchi" [LI 11, left] and "Zusanli" [ST 36, left]) in the non-target point group; and ribavirin solution, 120 µL was injected intraperitoneally in the ribavirin group. The intervention was delivered once daily, for consecutive 7 days. Three parallel experiments were undertaken. The mean death rate and survival time were assessed in each group, the body mass and lung index were compared among groups. Using HE staining, the morphology of lung tissue was observed; and with real-time fluorescence quantitative PCR, viral load in lung tissue was detected. The concentrations of inflammatory factors (tumor necrosis factor α [TNF-α], interleukin [IL]-1ß, IL-10) were detected in lung tissue of each group using ELISA; and those of oxidative stress markers (superoxide dismutase [SOD], glutathione peroxidase [GSH-Px], malondialdehyde [MDA]) were detected with chemiluminescence method. RESULTS: Compared with the control group, the body mass was decreased and lung index was increased in the model group (P<0.01). In comparison with the model group, body mass was increased in the acupoint injection group (P<0.05), lung index was reduced in the acupoint injection group the and ribavirin group (P<0.05); the mean death rate was decreased and the mean survival time prolonged in the mice of the acupoint injection group (P<0.01, P<0.05); and the mean death rate was reduced in the mice of the ribavirin group (P<0.05). In the model group, the alveolar structure was not integral, the alveolar septum was thickened, inflammatory cells were infiltrated and red blood cells exudated seriously (P<0.01). Compared with the model group, in the acupoint injection group and the ribavirin group, the alveolar structure was integral, the thickened alveolar septum was alleviated; and the infiltration of inflammatory cells and the exudation of red blood cells were reduced remarkably. The viral load was reduced in the mice of the ribavirin group when compared with the model group (P<0.01). Compared with the control group, the concentrations of TNF-α, IL-1ß and MDA in lung tissue were increased and those of IL-10, SOD and GSH-Px were reduced in the model group (P<0.01). In the acupoint injection group and the ribavirin group, the concentrations of TNF-α, IL-1ß and MDA were reduced in lung tissue and those of IL-10, SOD and GSH-Px were increased (P<0.05, P<0.01) when compared with the model group. CONCLUSION: Acupoint injection with VC+VBCo solution may alleviate inflammatory responses and oxidative stress in lung tissue of the PR8-induced pneumonia mice, improve survival rate and prolong the survival time in the case of no effect of the viral load.

Screening of non-alcoholic steatohepatitis (NASH)-related datasets and identification of NASH-related genes.

Non-alcoholic steatohepatitis (NASH) is a common liver disease in the western world. The mechanisms behind NASH formation are poorly understood, but there may be multiple targets considering the disease's multifactorial nature. To explore the genes related to the pathogenesis of NASH, we downloaded clinical data and gene expression of NASH patients from the Gene Expression Omnibus database (GEO). We identified 281 genes with a common expression in two NASH-related datasets (GSE89632 and GSE83452), suggesting that they may be related to NASH. Further study showed that Angptl4, Foxo1, and Ttc39B might be essential for NASH progression, and these have been poorly studied. Therefore, we explored their roles in NASH. Our data show that these genes participate in the development of NASH through lipid metabolism. This suggests that the three genes can be used as therapeutic targets in NASH.

miR-485-3p regulated by MALAT1 inhibits osteosarcoma glycolysis and metastasis by directly suppressing c-MET and AKT3/mTOR signalling.

AIMS: Osteosarcoma (OS) is an extremely malignant bone cancer with high incidence and rapid progression. This study aims to investigate the role and underlying mechanisms of MALAT1 and miR-485-3p in OS. MATERIALS AND METHODS: qRT-PCR and Western blotting were utilized to measure the levels of miR-485-3p, MALAT1, c-MET, AKT3, p-mTOR, mTOR, glycolysis-related proteins or migration-related proteins. Colony formation and transwell assay were used to test the roles of miR-485-3p, MALAT1, c-MET and AKT3 in cancer cell proliferation, migration and invasion. Dual luciferase assay was used to validate the interactions of miR-485-3p/c-MET, miR-485-3p/AKT3, and MALAT1/miR-485-3p. Glucose uptake assay and measurement of lactate production were employed to determine the glycolysis process. Mouse tumour xenograft model was used to determine the effect of shMALAT1 and miR-485-3p mimics on tumour growth and metastasis in vivo. KEY FINDINGS: miR-485-3p was decreased while c-MET, AKT3, and MALAT1 were increased in human OS tissues and cells. miR-485-3p bound directly to c-MET and AKT3 mRNAs and repressed OS cell glycolysis, proliferation, migration, and invasion through decreasing glycolysis-related proteins and migration-related proteins via inhibiting c-MET and AKT3/mTOR pathway. In addition, MALAT1 interacted with miR-485-3p and disinhibited c-MET and AKT3/mTOR signalling. Knockdown MALAT1 or overexpression of miR-485-3p restrained OS tumour growth and lung metastasis in vivo. SIGNIFICANCE: miR-485-3p suppresses OS glycolysis, proliferation, and metastasis via inhibiting c-MET and AKT3/mTOR signalling and MALAT1 acts as a sponge of miR-485-3p. MALAT1 and miR-485-3p may be the key regulators in OS progression, and potential molecular targets for future OS therapy.

[Acupuncture reduce colonic inflammation by suppressing oxidative stress and endoplasmic reticulum stress in rats with ulcerative colitis].

OBJECTIVE: To investigate the effect of manual acupuncture (MA) and electroacupuncture (EA) on histopathological changes, and levels of oxidative-stress related cytokines and key proteins of the endoplasmic reticulum stress (ERS) pathway in ulcerative colitis (UC) rats, so as to reveal their mechanisms underlying improvement of UC. METHODS: Twenty-eight male SD rats were randomly divided into control, model, EA and MA groups (n＝ 7 rats per group). The UC model was established by enema of mixture solution of 5% 2, 4, 6-trinitrobenzene sulfonic acid (TNBS, 100 mg/kg). Rats of the control group received intra-rectal perfusion of normal saline. After modeling, the left "Quchi"(LI11) and "Zusanli"(ST36) were stimulated with EA (2-4 mA，8 Hz/25 Hz) or MA for 20 min, once every other day for consecutive 2 weeks. The rats in the control and model group were just anesthetized and fixed. At the end of experiments, the colon tissue was collected for observing histopathological changes with H．E. staining. The contents of oxidative stress-related factors as superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), reduced glutathione (GSH) and total antioxidant capacity (T-AOC) were detected by ELISA, and the expression levels of key proteins of ERS as phosphorylated inhibitor of nuclear factor kappa B kinase α (p-IκBα), phosphorylated p65 (p-p65), glucose regulated protein 78 (GRP78), phosphorylated protein kinase R-like endoplasmic reticulum kinase (p-PERK) and phosphorylated eukaryotic translation initiation factor 2α (p-eIF2α) by using Western blot. RESULTS: After modeling, the colon tissues showed severe swelling, disordered arrangement of intestinal mucosal cells, hemorrhage with infiltration of inflammatory cell and partial loss of colon villus, which was relatively milder in the EA and MA groups. The colonic lesion score was remarkably increased in the model group in contrast to the control group (P<0.01), and obviously reduced in both EA and MA groups relevant to the model group (P<0.01). The levels of SOD, CAT, GSH and T-AOC were all significantly decreased (P<0.01), and the content of MDA, and expression levels of p-IκBα， p-p65 and GRP78, p-PERK and p-eIF2α proteins were all significantly increased in the model group relevant to the control group (P<0.01). After the treatment, modeling-induced down-regulation of SOD, CAT and GSH in both EA and MA groups, and T-AOC in the EA group, and up-regulation of levels of MDA, p-IκBα， p-p65, GRP78, p-PERK and p-eIF2α in both groups were reversed (P<0.01, P<0.05). CONCLUSION: Both EA and MA treatment can obviously alleviate colonic inflammation in UC rats via inhibiting oxidative stress and ERS.

[Protective effects of artesunate against Con A-induced autoimmune liver injury in mice].

Autoimmune liver disease is a refractory disease clinically, and there is no particularly effective drug at present. Therefore, it is of important clinical value to develop new effective intervention drugs for the prevention and treatment of autoimmune liver disease. In order to investigate the potential protective effect of artesunate (Art) on concanavalin A (Con A)-induced autoimmune liver injury, different doses of Art (27, 54, 108 mg·kg⁻¹) were orally administered to mice for consecutive 7 days, respectively. Then the Con A was injected into mice via tail vein to induce liver injury models. 8 h after modeling, the mice were sacrificed. The serum and liver tissue were collected for detecting the level of alanine aminotransferase (ALT), and aspartate transaminase (AST), liver pathological histopathology, inflammatory cytokines and nuclear factor (NF-κB) key protein expression level. The results showed that 108 mg·kg⁻¹ Art remarkably reduced Con A-induced liver indexes and serum transaminase levels (ALT and AST) as compared with model group(P<0.01). Meanwhile, the liver histopathological changes were obviously alleviated with a significant decrease of pro-inflammatory cytokine including tumor necrosis factor (TNF-α), interferon (IFN-γ), interleukin (IL-6), IL-17 and a higher increase of anti-inflammatory cytokine IL-10 (P<0.01). Western blot results showed that 108 mg·kg⁻¹ Art markedly inhibited the expressions of p-p65 and p-IκBα proteins (P<0.01). The specific inhibitor of NF-κB, pyrrolidinedithiocarbamate (PDTC) could also significantly inhibit the expressions of p-p65 and p-IκBα with and alleviate liver injuries. Therefore, our results indicated that Art may have a protective action against Con A-induced autoimmune liver injury mainly by suppressing NF-κB signal pathway in mice. The study provides scientific reference for artesunate usage in preventing autoimmune liver injury.

[Effect of Electroacupuncture and Manual Acupuncture on Colonic Inflammatory Injury, Cytokine Levels and Cell Apoptosis in Ulcerative Colitis Rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) and manual acupuncture interventions on colonic inflammatory injury, cytokine level and cell apoptosis in ulcerative colitis (UC)rats, so as to reveal its mechanisms underlying improvement of UC. METHODS: A total of 32 SD rats were randomized into control, model, EA and manual acupuncture groups (8 rats/group). The UC model was established by intrarectally administration of 5% trinitro-benzene-sulfonic acid (TNBS)+ dehydrated alcohol. Both "Quchi" (LI 11) and "Zusanli" (ST 36) were punctured with filiform needles for 20 min in the manual acupuncture group or stimulated with EA (8 Hz/25 Hz, 2-4 mA, and duration of 20 min) in the EA group. The treatment was conducted once daily for consecutive 6 days. Changes of body weight and pathological state of colitis were observed. The contents of pro-inflammatory cytokines (TNF-α, IL-1 ß, IL-6), anti-inflammatory cytokine (IL-10), homocysteine (Hcy) and myeloperoxidase (MPO, two oxygen free radicals associated substances) in the colon tissues were detected by ELISA, and the protein expression levels of Bcl-2,Bax,phosphorylated (p)-inhibitor of nuclear factor kappaB kinase α(IκBα) and p-p 65 (a subunit of nuclear factor) of colonic tissues were detected by Western blot, separately. RESULTS: Compared with the control group, the body weight was decreased and the state of the swelling and hemorrhage of the colon got worsened in the model group, while the state of the swelling and hemorrhage of the colon was better in both EA and manual acupuncture groups, and the body weight was clearly increased from day 4 on in both treatment groups. The concentrations of colonic TNF-α, IL-1 ß, IL-6, IL-10, MPO and Hcy were all significantly higher in the model group than in the control group (P<0.01), but those of colonic TNF-α, IL-1 ß, IL-6 in both EA and manual acupuncture groups, those of MPO and Hcy in the EA group were significantly down-regulated following the intervention (P<0.05, P<0.01), and that of IL-10 was notably further increased in the manual acupuncture group (P<0.05). In addition, modeling-induced remarkable down-regulation of colonic Bcl-2/Bax, and marked up-regulation of expression of IκBα and p-p 65 proteins were significantly suppressed in both EA and manual acupuncture groups (P<0.01). CONCLUSIONS: Both EA and manual acupuncture interventions Feb alleviate the colonic lesions in UC rats, which Feb be related to their functions in regulating levels of pro-inflammatory and anti-inflammatory cytokines, in balancing the expression of apoptosis-related protein and anti-apoptosis-related protein and in down-regulating the expression of the key nuclear transcription factors in the colonic tissue.

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.

[Biomechanical study on a net-fixation of Kirschner wire in treating depressed tibial plateau fractures].

OBJECTIVE: To evaluate the biomechanical properties of tibial plateau depressed fracture fixed with a net-fixation of Kirschner wires. METHODS: Twenty homemade fracture models were fixed with eight 1.5 mm Kirschner wires in a net-fixation; 20 homemade fracture models were fixed with two 3.5 mm cortical screws. Plane-compressed and dot-compressed test were made on each 10 models of the two groups. The maximal force of anti-ompress and stiffness were measured and evaluated. RESULTS: In plane-compressed test,mean maximal force of anti-compress and stiffness for screw fixation was (1,925.31 +/- 444.26) N and (2.28 +/- 0.53) N/mm2, respectively, for net-fixation was (1,609.62 +/- 277.72) N and (1.90 +/- 0.33) N/mm2, respectively. There was no statistical difference between the two fixation methods (P > 0.05). In dot-compressed test,mean maximal force of anti-compress and stiffness for screw fixation was (411.13 +/- 233.88) N and (2.66 +/- 1.52) N/mm2,respectively,for net-fixation was (1,105.58 +/- 290.66) N and (7.18 +/- 1.89) N/mm2,respectively,the net-fixation was better than that of the screw fixation (P< 0.01). CONCLUSION: Treatment of tibial plateau depressed fracture with a net-fixation of Kirschner wires is a biological fixation and is a reliably method.

[Effect of the melamine residue in soil on growth of Chinese cabbage].

Soil and foliar application of melamine (ME) treatments to 'Zaoshu 5' Chinese cabbage were investigated. The ME was degraded very slowly in soil treated with different dosages (40,160 and 800 mg x kg(-1)), and 90 days later the residuals of ME were 21.1%, 15.8% and 43.6% respectively. The Chinese cabbage could take in exogenously applied ME through its root and stem leaf and accumulate it to considerable levels with the increasing applied density. In soil application test, the maximum and minimum contents of ME were 105.7 and 8.0 mg x kg(-1) in root, and 139.9 and 7.1 mg x kg(-1) in stem leaf; the ME transport occurred from root to stem leaf. In foliar application test,the maximum and minimum contents of ME were 4.3 and 0.9 mg x kg(-1) in root, and 8.5 and 3.2 mg x kg(-1) in stem leaf. In soil application test,the low level of ME (40 mg x kg(-1)) increased the biomass yield by 9.8% and the high level of ME (800 mg x kg(-1)) decreased the biomass yield by 15.9%; the contents of chlorophyll and soluble sugar increased,but the content of Vitamin C decreased. Foliar application ME had no obvious significance on the growth of Chinese cabbage. The studies indicate that the residual time of ME in soil is long and the Chinese cabbage can absorb exogenously applied ME and ME can affect the growth of Chinese cabbage.

[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.