Mechanism of Reactive Oxygen/Nitrogen Species in Liver Ischemia-Reperfusion Injury and Preventive Effect of Chinese Medicine.

Liver ischemia-reperfusion injury (LIRI) is a pathological process involving multiple injury factors and cell types, with different stages. Currently, protective drugs targeting a single condition are limited in efficacy, and interventions on immune cells will also be accompanied by a series of side effects. In the current bottleneck research stage, the multi-target and obvious clinical efficacy of Chinese medicine (CM) is expected to become a breakthrough point in the research and development of new drugs. In this review, we summarize the roles of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in various stages of hepatic ischemia-reperfusion and on various types of cells. Combined with the current research progress in reducing ROS/RNS with CM, new therapies and mechanisms for the treatment of hepatic ischemia-reperfusion are discussed.

Naringin ameliorates liver fibrosis in zebrafish by modulating IDO1-mediated lipid metabolism and inflammatory infiltration.

Liver fibrosis (LF) is an important reparative process in response to acute or chronic hepatic injury, which has the potential to advance towards cirrhosis and hepatocellular carcinoma. Dietary naringin consumption contributes to protection against LF in animal studies, while the exact protective mechanism of naringin remains unclear. This study aimed to investigate the molecular mechanisms behind the potential protective effect of naringin against TAA-induced LF in zebrafish. In this study, we utilized zebrafish to create the LF model and investigate the therapeutic mechanism of naringin. Firstly, we evaluated the changes in hepatic fibrosis and lipid accumulation in the liver following naringin treatment with oil red O, Nile red, and Sirius red and immunohistochemistry. In addition, we employed an ROS probe to directly measure oxidative stress and monitor inflammatory cell migration in a zebrafish transgenic line. Morpholino was used in the knockdown of IDO1 in order to verify its vital role in LF. Our findings demonstrated that naringin exhibited anti-inflammatory and anti-fibrotic action in conjunction with a reversal in lipid accumulation, oxidative stress and suppression of macrophage infiltration and activation of hepatic stellate cells. Furthermore, the results showed that the antifibrotic effect of naringin was removed upon IDO1 knockdown, proving that naringin exerts a protective effect by regulating IDO1. Naringin demonstrates remarkable protective effects against LF, effectively counteracting inflammation and hepatic steatosis in zebrafish liver. These findings suggest that naringin may function as an effective IDO1 inhibitor, holding the potential for clinical translation as a therapeutic agent for the treatment of LF.

Caveolin-1 is critical for hepatic iron storage capacity in the development of nonalcoholic fatty liver disease.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is associated with disordered lipid and iron metabolism. Our previous study has substantiated the pivotal role of Caveolin-1 (Cav-1) in protecting hepatocytes and mediating iron metabolism in the liver. This study aimed to explore the specific mechanisms underlying the regulation of iron metabolism by Cav-1 in NAFLD. METHODS: Hepatocyte-specific Cav-1 overexpression mice and knockout mice were used in this study. Cav-1-knockdown of RAW264.7 cells and mouse primary hepatocytes were performed to verify the changes in vitro. Moreover, a high-fat diet and palmitic acid plus oleic acid treatment were utilized to construct a NAFLD model in vivo and in vitro, respectively, while a high-iron diet was used to construct an in vivo iron overload model. Besides, iron concentration, the expression of Cav-1 and iron metabolism-related proteins in liver tissue or serum were detected using iron assay kit, Prussian blue staining, Western blotting, immunofluorescence staining, immunohistochemical staining and ELISA. The related indicators of lipid metabolism and oxidative stress were evaluated by the corresponding reagent kit and staining. RESULTS: Significant disorder of lipid and iron metabolism occurred in NAFLD. The expression of Cav-1 was decreased in NAFLD hepatocytes (P < 0.05), accompanied by iron metabolism disorder. Cav-1 enhanced the iron storage capacity of hepatocytes by activating the ferritin light chain/ferritin heavy chain pathway in NAFLD, subsequently alleviating the oxidative stress induced by excess ferrous ions in the liver. Further, CD68+CD163+ macrophages expressing Cav-1 were found to accelerate iron accumulation in the liver, which was contrary to the effect of Cav-1 in hepatocytes. Positive correlations were also observed between the serum Cav-1 concentration and the serum iron-related protein levels in NAFLD patients and healthy volunteers (P < 0.05). CONCLUSIONS: These findings confirm that Cav-1 is an essential target protein that regulates iron and lipid metabolic homeostasis. It is a pivotal molecule for predicting and protecting against the development of NAFLD.

Guanidinoacetic Acid Attenuates Adipogenesis through Regulation of miR-133a in Sheep.

Guanidinoacetic acid (GAA) is an amino acid derivative, previously described in the skeletal muscle of vertebrates, that serves as an important regulator of cellular bioenergetics and has been widely used as a feed additive. Nevertheless, the effect of GAA on adipose tissue growth remains unclear. Here, we hypothesized that dietary GAA negatively affected adipose tissue development in lambs. Lambs were individually fed diets with (0.09%) or without GAA for 70 d ad libitum, and the subcutaneous adipose tissues were sampled for analysis. The results showed that dietary GAA supplementation decreased the girth rib (GR) value (p < 0.01) of lamb carcasses. Both real-time PCR and Western blot analysis suggested that dietary GAA inhibited the expression of adipogenic markers, including peroxisome proliferator-activated receptor γ (PPARγ, p < 0.05), CCAAT/enhancer-binding protein α (C/EBPα, p < 0.01) and sterol-regulatory-element-binding protein 1c (SREBP1C, p < 0.01) in subcutaneous adipose tissue. In vitro, GAA inhibited sheep stromal vascular fraction (SVF) cell proliferation, which was associated with downregulation of proliferating cell nuclear antigen (PCNA, p < 0.05), cyclin-dependent kinase 4 (CDK 4, p < 0.05) and cyclin D1 (p < 0.01). GAA suppressed adipogenesis of SVF cells. Furthermore, miRNA sequencing revealed that GAA affected the miRNA expression profile, and real-time PCR analysis confirmed that miR-133a expression in both subcutaneous adipose tissue and SVF cell was downregulated by GAA. Meanwhile, miR-133a promoted adipogenic differentiation of SVF cells by targeting Sirt1. miR-133a mimics alleviated the inhibitory effect of GAA on SVF cells' adipogenic differentiation. In summary, GAA attenuated adipogenesis of sheep SVF cells, which might occur through miR-133a-modulated Sirt1 expression.

Lip morphology estimation models based on three-dimensional images in a modern adult population from China.

Lips are the main part of the lower facial soft tissue and are vital to forensic facial approximation (FFA). Facial soft tissue thickness (FSTT) and linear measurements in three dimensions are used in the quantitative analysis of lip morphology. With most FSTT analysis methods, the surface of soft tissue is unexplicit. Our study aimed to determine FSTT and explore the relationship between the hard and soft tissues of lips in different skeletal occlusions based on cone-beam CT (CBCT) and 3dMD images in a Chinese population. The FSTT of 11 landmarks in CBCT and 29 lip measurements in CBCT and 3dMD of 180 healthy Chinese individuals (90 males, 90 females) between 18 and 30 years were analyzed. The subjects were randomly divided into two groups with different skeletal occlusions distributed equally: 156 subjects in the experimental group to establish the prediction regression formulae of lip morphology and 24 subjects in the test group to assess the accuracy of the formulae. The results indicated that FSTT in the lower lip region varied among different skeletal occlusions. Furthermore, sex discrepancy was noted in the FSTT in midline landmarks and linear measurements. Measurements showing the highest correlation between soft and hard tissues were between total upper lip height and Ns-Pr (0.563 in males, 0.651 in females). The stepwise multiple regression equations were verified to be reliable with an average error of 1.246 mm. The method of combining CBCT with 3dMD provides a new perspective in predicting lip morphology and expands the database for FFA.

Three-dimensional superimposition of digital models for individual identification.

Dentition is an individualizing structure in humans that may be potentially utilized in individual identification. However, research on the use of three-dimensional (3D) digital models for personal identification is rare. This study aimed to develop a method for individual identification based on a 3D image registration algorithm and assess its feasibility in practice. Twenty-eight college students were recruited; for each subject, a dental cast and an intraoral scan were taken at different time points, and digital models were acquired. The digital models of the dental casts and intraoral scans were assumed as antemortem and postmortem dentition, respectively. Additional 72 dental casts were extracted from a hospital database as a suspect pool together with 28 antemortem models. The dentition images of all of the models were extracted. Correntropy was introduced into the traditional iterative closest point algorithm to compare each postmortem 3D dentition with 3D dentitions in the suspect pool. Point-to-point root mean square (RMS) distances were calculated, and then 28 matches and 2772 mismatches were obtained. Statistical analysis was performed using the Mann-Whitney U test, which showed significant differences in RMS between matches (0.18±0.03mm) and mismatches (1.04±0.67mm) (P<0.05). All of the RMS values of the matched models were below 0.27mm. The percentage of accurate identification reached 100% in the present study. These results indicate that this method for individual identification based on 3D superimposition of digital models is effective in personal identification.

Three-dimensional prediction of nose morphology in Chinese young adults: a pilot study combining cone-beam computed tomography and 3dMD photogrammetry system.

The nose is the most prominent part of the face and is a crucial factor for facial esthetics as well as facial reconstruction. Although some studies have explored the features of external nose and predicted the relationships between skeletal structures and soft tissues in the nasal region, the reliability and applicability of methods used in previous studies have not been reproduced. In addition, the majority of previous studies have focused on the sagittal direction, whereas the thickness of the soft tissues was rarely analyzed in three dimensions. A few studies have explained the specific characteristics of the nose of Chinese individuals. The aim of this study was to investigate the relationship between the hard nasal structures and soft external nose in three dimensions and to predict the morphology of the nose based on hard-tissue measurements. To eliminate the influence of low resolution of CBCT and increase the accuracy of measurement, three-dimensional (3D) images captured by cone-beam computed tomography (CBCT) and 3dMD photogrammetry system were used in this study. Twenty-six measurements (15 measurements for hard tissue and 11 measurements for soft tissue) based on 5 craniometric and 5 capulometric landmarks of the nose of 120 males and 120 females were obtained. All of the subjects were randomly divided into an experimental group (180 subjects consisting of 90 males and 90 females) and a test group (60 subjects consisting of 30 males and 30 females). Correlation coefficients between hard- and soft-tissue measurements were analyzed, and regression equations were obtained based on the experimental group and served as predictors to estimate nasal morphology in the test group. Most hard- and soft-tissue measurements appeared significantly different between genders. The strongest correlation was found between basis nasi protrusion and nasospinale protrusion (0.499) in males, and nasal height and nTr-nsTr (0.593) in females. For the regression equations, the highest value of R2 was observed in the nasal bridge length in males (0.257) and nasal tip protrusion in females (0.389). The proportion of subjects with predicted errors < 10% was over 86.7% in males and 70.0% in females. Our study proved that a combined CBCT and 3dMD photogrammetry system is a reliable method for nasal morphology estimation. Further research should investigate other influencing factors such as age, skeletal types, facial proportions, or population variance in nasal morphology estimation.

[Effect of Dry Needling Stimulation of Myofascial Trigger Point on Sample Entropy of Electromyography of Gastrocnemius Injured Site in Rats].

OBJECTIVE: To attempt to establish an objective quantitative indicator to characterize the trigger point activity, so as to evaluate the effect of dry needling on myofascial trigger point activity. METHODS: Twenty-four male Sprague-Dawley rats were randomly divided into blank control group, dry needling (needling) group, stretching exercise (stretching) group and needling plus stretching group (n＝6 per group). The chronic myofascial pain (trigger point) model was established by freedom vertical fall of a wooden striking device onto the mid-point of gastrocnemius belly of the left hind-limb to induce contusion, followed by forcing the rat to make a continuous downgrade running exercise at a speed of 16 m/min for 90 min on the next day which was conducted once a week for 8 weeks. Electromyography (EMG) of the regional myofascial injured point was monitored and recorded using an EMG recorder via electrodes. It was considered success of the model if spontaneous electrical activities appeared in the injured site. After a 4 weeks' recovery, rats of the needling group were treated by filiform needle stimulation (lifting-thrusting-rotating) of the central part of the injured gastrocnemius belly (about 10 mm deep) for 6 min, and those of the stretching group treated by holding the rat's limb to make the hip and knee joints to an angle of about 180°, and the ankle-joint about 90° for 1 min every time, 3 times altogether (with an interval of 1 min between every 2 times). The activity of the trigger point was estimated by the sample entropy of the EMG signal sequence in reference to Richman's and Moorman's methods to estimate the curative effect of both needling and exercise. RESULTS: After the modeling cycle, the mean sample entropies of EMG signals was significantly decreased in the model groups (needling group [0.034±0.010], stretching group [0.045±0.023], needling plus stretching group [0.047±0.034]) relevant to the blank control group (0.985±0.196, P<0.01). After the treatment, the mean sample entropy of EMG signals was evidently increased in both needling (0.819±0.088), stretching (0.532±0.25) and needling plus stretching (0.810±0.117) groups (P<0.01). The mean sample entropy of the needling and needling plus stretching groups were significantly higher than that of the stretching group (P<0.01), without remarkable difference between the two needling groups in the mean sample entropy (P>0.05), suggesting a better efficacy of dry needling in easing trigger point activity. CONCLUSION: Dry needling is able to relieve myofascial trigger point activity in rats, which is better than that of simple passive stretching therapy.

Evidence for Dry Needling in the Management of Myofascial Trigger Points Associated With Low Back Pain: A Systematic Review and Meta-Analysis.

OBJECTIVE: To evaluate the current evidence of the effectiveness of dry needling of myofascial trigger points (MTrPs) associated with low back pain (LBP). DATA SOURCES: PubMed, Ovid, EBSCO, ScienceDirect, Web of Science, Cochrane Library, CINAHL, and China National Knowledge Infrastructure databases were searched until January 2017. STUDY SELECTION: Randomized controlled trials (RCTs) that used dry needling as the main treatment and included participants diagnosed with LBP with the presence of MTrPs were included. DATA EXTRACTION: Two reviewers independently screened articles, scored methodologic quality, and extracted data. The primary outcomes were pain intensity and functional disability at postintervention and follow-up. DATA SYNTHESIS: A total of 11 RCTs involving 802 patients were included in the meta-analysis. Results suggested that compared with other treatments, dry needling of MTrPs was more effective in alleviating the intensity of LBP (standardized mean difference [SMD], -1.06; 95% confidence interval [CI], -1.77 to -0.36; P=.003) and functional disability (SMD, -0.76; 95% CI, -1.46 to -0.06; P=.03); however, the significant effects of dry needling plus other treatments on pain intensity could be superior to dry needling alone for LBP at postintervention (SMD, 0.83; 95% CI, 0.55-1.11; P<.00001). CONCLUSIONS: Moderate evidence showed that dry needling of MTrPs, especially if associated with other therapies, could be recommended to relieve the intensity of LBP at postintervention; however, the clinical superiority of dry needling in improving functional disability and its follow-up effects still remains unclear.

Decreased Spontaneous Electrical Activity and Acetylcholine at Myofascial Trigger Spots after Dry Needling Treatment: A Pilot Study.

OBJECTIVE: The aims of this study are to investigate the changes in spontaneous electrical activities (SEAs) and in acetylcholine (ACh), acetylcholine receptor (AChR), and acetylcholine esterase (AChE) levels after dry needling at myofascial trigger spots in model rats. MATERIALS AND METHODS: Forty-eight male Sprague-Dawley rats were divided into four groups. Thirty-six rats were assigned to three model groups, which underwent MTrSs modeling intervention. Twelve rats were assigned to the blank control (BC) group. After model construction, the 36 model rats were randomly subdivided into three groups according to treatment: MTrSs model control (MC) and two dry needling groups. One dry needling group received puncturing at MTrSs (DN-M), whereas the other underwent puncturing at non-MTrSs (DN-nM). Dry needling treatment will last for two weeks, once a week. SEAs and ACh, AChR, and AChE levels were measured after one-week rest of dry needling treatment. RESULTS: The amplitudes and frequencies of endplate noise (EPN) and endplate spike (EPS) significantly decreased after dry needling treatment in the DN-M group. Moreover, ACh and AChR levels significantly decreased, whereas AChE significantly increased after dry needling treatment in the DN-M group. CONCLUSION: Dry needling at the exact MTrSs is more effective than dry needling at non-MTrSs.