Feeding artery: a valuable feature for differentiation of regenerative nodule, dysplastic nodules and small hepatocellular carcinoma in CEUS LI-RADS.

OBJECTIVE: To investigate whether the feeding artery (FA) feature can aid in discriminating small hepatocellular carcinoma (HCC) using the contrast-enhanced ultrasound (CEUS) Liver Imaging Reporting and Data System (LI-RADS) from precancerous lesions. METHODS: Between June 2017 and May 2021, a total of 347 patients with 351 precancerous liver lesions or small HCCs who underwent CEUS were enrolled. Two independent radiologists assigned LI-RADS categories to all lesions and assessed the presence of the FA feature, which was used as an ancillary feature to either upgrade or downgrade the LI-RADS category. The diagnostic performance of CEUS LI-RADS, both with and without the FA feature, was evaluated based on accuracy, sensitivity, specificity, positive predictive value, and negative predictive value. RESULTS: The FA feature was found to be more prevalent in HCC (85.54%, p < 0.001) than in regenerative nodules (RNs, 29.73%), low-grade dysplastic nodules (LGDNs, 33.33%), and high-grade dysplastic nodules (HGDNs, 55.26%). Furthermore, the presence of arterial phase hyperenhancement (APHE), washout (WO), and FA in liver nodules was associated with a higher expression of GPC-3 and Ki-67 compared to the group without these features (p < 0.001). After adjusting, the sensitivity and accuracy of LR-5 for HCC improved from 68.67% (95%CI: 62.46%, 74.30%) to 77.51% (95%CI: 71.72%, 82.44%) and from 69.23% (95%CI: 64.11%, 74.02%) to 73.79% (95%CI: 68.86%, 78.31%), respectively. CONCLUSION: The FA feature is a valuable feature for distinguishing small HCC and precancerous lesions and could be added as a possible ancillary feature in CEUS LI-RADS which was backed up by biomarkers. CLINICAL RELEVANCE STATEMENT: The presence of a feeding artery is a valuable imaging feature in the differentiation of HCC and precancerous lesions. Incorporating this characteristic in the CEUS LI-RADS can enhance the diagnostic ability. KEY POINTS: • Feeding artery is more frequent in HCC than in regenerative nodules, low-grade dysplastic nodules, and high-grade dysplastic nodules. • Feeding artery feature is a valuable ancillary feature for CEUS LI-RADS to differentiate regenerative nodules, low-grade dysplastic nodules, high-grade dysplastic nodules, and HCC. • The existence of feeding artery, arterial phase hyperenhancement, and washout is associated with more GPC-3 positive expression and higher Ki-67 expression than the group without these features.

On-Site Diagnostic Ability of CEUS/CT/MRI for Hepatocellular Carcinoma (2019-2022): A Multicenter Study.

OBJECTIVES: To compare the on-site diagnostic performance of contrast-enhanced ultrasound (CEUS), computed tomography (CECT), and magnetic resonance imaging (CEMRI) for hepatocellular carcinoma (HCC) across diverse practice settings. METHODS: Between May 2019 and April 2022, a total of 2085 patients with 2320 pathologically confirmed focal liver lesions (FLLs) were enrolled. Imaging reports were compared with results from pathology analysis. Diagnostic performance was analyzed in defined size, high-risk factors for HCC, and hospital volume categories. RESULTS: Three images achieved similar diagnostic performance in classifying HCC from 16 types of FLLs, including HCC ≤2.0 cm. For HCC diagnosis at low-volume hospitals and HCC with high-risk factors, the accuracy and specificity of CEUS were comparable to CECT and CEMRI, while the sensitivity of CEUS (77.4 and 89.5%, respectively) was inferior to CEMRI (87.0 and 92.8%, respectively). The diagnostic accuracy of CEUS + CEMRI and CEUS + CECT increased by 7.8 and 6.2% for HCC ≤2.0 cm, 8.0 and 5.0% for HCC with high-risk factors, and 7.4 and 5.5% for HCC at low-volume hospitals, respectively, compared with CEMRI/CECT alone. CONCLUSIONS: Compared with CECT and CEMRI, CEUS provides adequate diagnostic performance in clinical first-line applications at high-volume hospitals. Moreover, a higher diagnostic performance for HCC is achieved by combining CEUS with CECT/CEMRI compared with any single imaging technique.

Ultrasound identification of hepatic echinococcosis using a deep convolutional neural network model in China: a retrospective, large-scale, multicentre, diagnostic accuracy study.

BACKGROUND: Ultrasonography is the most widely used technique to diagnose echinococcosis; however, challenges in using this technique and the demand on medical resources, especially in low-income or remote areas, can delay diagnosis. We aimed to develop a deep convolutional neural network (DCNN) model based on ultrasonography to identify echinococcosis and its types, especially alveolar echinococcosis. METHODS: This retrospective, large-scale, multicentre study used ultrasound images from patients assessed at 84 hospitals in China, obtained between Jan 1, 2002, and Dec 31, 2021. Patients with a diagnosis of cystic echinococcosis, alveolar echinococcosis, or seven other types of focal liver lesions were included. We tested ResNet-50, ResNext-50, and VGG-16 as the backbone network architecture for a classification DCNN model and input the perinodular information from the ultrasound images. We trained and validated the DCNN model to diagnose and classify echinococcosis using still greyscale ultrasound images of focal liver lesions in four stages: differentiating between echinococcosis and other focal liver lesions (stage one); differentiating cystic echinococcosis, alveolar echinococcosis, and other focal liver lesions (stage two); differentiating cystic echinococcosis, alveolar echinococcosis, benign other focal liver lesions, and malignant focal liver lesions (stage three); and differentiating between active and transitional cystic echinococcosis and inactive cystic echinococcosis (stage four). We then tested the algorithm on internal, external, and prospective test datasets. The performance of DCNN was also compared with that of 12 radiologists recruited between Jan 15, 2022, and Jan 28, 2022, from Qinghai, Xinjiang, Anhui, Henan, Xizang, and Beijing, China, with different levels of diagnostic experience for echinococcosis and other focal liver lesions in a subset of ultrasound data that were randomly chosen from the prospective test dataset. The study is registered at ClinicalTrials.gov (NCT03871140). FINDINGS: The study took place between Jan 1, 2002, and Dec 31, 2021. In total, to train and test the DCNN model, we used 9631 liver ultrasound images from 6784 patients (2819 [41·7%] female patients and 3943 [58·3%] male patients) from 87 Chinese hospitals. The DCNN model was trained with 6328 images, internally validated with 984 images, and tested with 2319 images. The ResNet-50 network architecture outperformed VGG-16 and ResNext-50 and was generalisable, with areas under the receiver operating characteristic curve (AUCs) of 0·982 (95% CI 0·960-0·994), 0·984 (0·972-0·992), and 0·913 (0·886-0·935) in distinguishing echinococcosis from other focal liver lesions; 0·986 (0·966-0·996), 0·962 (0·946-0·975), and 0·900 (0·872-0·924) in distinguishing alveolar echinococcosis from cystic echinococcosis and other focal liver lesions; and 0·974 (0·818-1·000), 0·956 (0·875-0·991), and 0·944 (0·844-0·988) in distinguishing active and transitional cystic echinococcosis from inactive echinococcosis in the three test datasets. Specifically, in patients with the hepatitis B or hepatitis C virus, the model could distinguish alveolar echinococcosis from hepatocellular carcinoma with an AUC of 0·892 (0·812-0·946). In identifying echinococcosis, the model showed significantly better performance compared with senior radiologists from a high-endemicity area (AUC 0·942 [0·904-0·967] vs 0·844 [0·820-0·866]; p=0·027) and improved the diagnostic ability of junior, attending, and senior radiologists before and after assistance with AI with comparison of AUCs of 0·743 (0·714-0·770) versus 0·850 (0·826-0·871); p<0·0001, 0·808 (0·782-0·832) versus 0·886 (0·864-0·905); p<0·0001, and 0·844 (0·820-0·866) versus 0·870 (0·847-0·890); p=0·092, respectively. INTERPRETATION: The DCNN model was shown to be accurate and robust, and could improve the ultrasound diagnostic ability of radiologists for echinococcosis and its types for highly endemic and remote regions. FUNDING: National Natural Science Foundation of China and National Key Research & Development Program of China. TRANSLATION: For the Chinese translation of the abstract see Supplementary Materials section.

Liver cirrhosis and tumor location can affect the range of intrahepatic microwave ablation zone.

BACKGROUND: The range of an ablation zone (AZ) plays a crucial role in the treatment effect of microwave ablation (MWA). The aim of this study was to analyze the factors influencing the AZ range. METHODS: Fourteen factors in four areas were included: patient-related factors (sex, age), disease-related factors (tumor location, liver cirrhosis), serological factors (ALT, AST, total protein, albumin, total bilirubin, direct bilirubin, and platelets), and MWA parameters (ablation time, power, and needle type). Multiple sequence MRI was used to delineate AZ by three radiologists using 3D Slicer. MATLAB was used to calculate the AZ length, width, and area of the largest section. Linear regression analysis was used to analyze influencing factors. Moreover, a subgroup analysis was conducted for patients with viral hepatitis. RESULT: 220 patients with 290 tumors were included between 2010-2021. In addition to MWA parameters, cirrhosis and tumor location were significant factors that influenced AZ (p < 0.001). The standardized coefficient (beta) of cirrhosis (cirrhosis vs. non-cirrhosis) was positive, which meant cirrhosis would lead to a decrease in AZ range. The beta of tumor location (near the hepatic hilar zone, intermediate zone, and periphery zone) was negative, indicating that AZ range decreased as the tumor location approached the hepatic hilum. For viral hepatitis patients, Fibrosis 4 (FIB4) score was a significant factor influencing AZ (p < 0.001), and the beta was negative, indicating that AZ range decreased as FIB4 increased. CONCLUSION: Liver cirrhosis, tumor location, and FIB4 affect the AZ range and should be considered when planning MWA parameters.

The tumor ghost on MRI after microwave ablation for hepatocellular carcinoma: A new modality to assess the ablative margin.

PURPOSE: The ablated tumor ghost can be visually distinguished on MR images after ablation. This retrospective study aimed to assess the performance of tumor ghost on post-ablation contrast-enhanced MRI with excellular contrast agent gadolinium-DTPA in evaluating the ablative margin of hepatocellular carcinoma (HCC) after microwave ablation (MWA). METHOD: 315 HCC lesions less than 5 cm in 287 patients completely treated by MWA were enrolled in the study. The tumor ghost was characterized as a lower signal intensity area than the surrounding tissues of the ablation zone on T1WI imaging. The ablation margin (AM) status was classified into AM0 (＞5mm) and AM1 (＜5mm) according to the minimum distance between the tumor ghost and ablated zone. Inter-observer agreement between two radiologists on the AM assessment was analyzed using the Cohen κ coefficient. Multivariate analysis using Cox proportional hazard model was performed to investigate independent risk factors for LTP. RESULTS: 175 and 140 tumors were evaluated as AM0 and AM1 through tumor ghost. The inter-observer agreement level between two radiologists for assessment of AM was good (κ coefficient = 0.752, 95 % confidence interval: 0.679-0.825, p < 0.001). The mediate follow-up period was 32.2 months (range 3.0-60.8 months). The incidence of LTP in the AM0 lesions and AM1 lesions was 6.3 % (11/175) and 20.0 % (28/140), respectively. AM status was identified as an independent prognostic factor for LTP (HR 3.057, 95 % CI, 1.445-6.470, p = 0.003). CONCLUSIONS: The assessment of the AM by tumor ghost on post-ablation MRI is an accurate and efficiently method for evaluating the completeness of microwave ablation for hepatocellular carcinoma.

Combination therapy using microwave ablation and d-mannose-chelated iron oxide nanoparticles inhibits hepatocellular carcinoma progression.

Despite being a common therapy for hepatocellular carcinoma (HCC), insufficient thermal ablation can leave behind tumor residues that can cause recurrence. This is believed to augment M2 inflammatory macrophages that usually play a pro-tumorigenic role. To address this problem, we designed d-mannose-chelated iron oxide nanoparticles (man-IONPs) to polarize M2-like macrophages into the antitumor M1 phenotype. In vitro and in vivo experiments demonstrated that man-IONPs specifically targeted M2-like macrophages and accumulated in peri-ablation zones after macrophage infiltration was augmented under insufficient microwave ablation (MWA). The nanoparticles simultaneously induced polarization of pro-tumorigenic M2 macrophages into antitumor M1 phenotypes, enabling the transformation of the immunosuppressive microenvironment into an immunoactivating one. Post-MWA macrophage polarization exerted robust inhibitory effects on HCC progression in a well-established orthotopic liver cancer mouse model. Thus, combining thermal ablation with man-IONPs can salvage residual tumors after insufficient MWA. These results have strong potential for clinical translation.

Radiomics analysis of ultrasound to predict recurrence of hepatocellular carcinoma after microwave ablation.

OBJECTIVE: To develop and validate an ultrasonic radiomics model for predicting the recurrence and differentiation of hepatocellular carcinoma (HCC). Convolutional neural network (CNN) ResNet 18 and Pyradiomics were used to analyze gray-scale-ultrasonic images to predict the prognosis and degree of differentiation of HCC. METHODS: This retrospective study enrolled 513 patients with HCC who underwent preoperative grayscale-ultrasonic imaging, and their clinical characteristics were observed. Patients were randomly divided into training (n = 413) and validation (n = 100) cohorts. CNN ResNet 18 and Pyradiomics were used to analyze ultrasonic images of HCC and peritumoral images to develop a prognostic and differentiation model. Clinical characteristics were integrated into the radiomics model and patients were stratified into high- and low-risk groups. The predictive effect was evaluated using the C-index and receiver operating characteristic (ROC) curve. RESULTS: The model combined with ResNet 18 and clinical characteristics achieved a good predictive ability. The C-indices of early recurrence (ER), late recurrence (LR), and recurrence-free survival (RFS) were 0.695 (0.561-0.789), 0.715 (0.623-0.800) and 0.721 (0.647-0.795), respectively, in the validation cohort, which was superior to the clinical model and ultrasonic semantic model. The model could stratify patients into high- and low-risk groups, which showed significant differences (p < 0.001) in ER, LR, and RFS. The area under the curve for predicting the degree of HCC differentiation was 0.855 and 0.709 in the training and validation cohorts, respectively. CONCLUSION: We developed and validated a radiomics model to predict HCC recurrence and HCC differentiation, which could also acquire pathological information in a noninvasive manner.KEY RESULTSA hepatocellular carcinoma (HCC) prognostic prediction model was developed and validated by convolutional neural network (CNN) ResNet 18-based gray-scale ultrasound (US).A differentiation of HCC prediction model was developed for preoperative prediction avoiding invasive operation.Compared with Pyradiomics, CNN ResNet was more suitable for extracting information from US images.

Engineered Probiotic Lactococcus lactis for Lycopene Production against ROS Stress in Intestinal Epithelial Cells.

Lactococcus lactis is a food-grade chassis for delivery of bioactive molecules to the intestinal mucosa in situ, while its ability to produce lycopene for detoxification of reactive oxidative species (ROS) is not realized yet. Here, L. lactis NZ9000 was engineered to synthesize lycopene by heterologous expression of a gene cluster crtEBI in plasmids or chromosomes, yielding the recombinant strains NZ4 and NZ5 with 0.59 and 0.54 mg/L lycopene production, respectively. To reroute the pyruvate flux to lycopene, the main lactate dehydrogenase and α-acetolactate synthase pathways were sequentially disrupted. The resultant strains NZΔldh-1 and NZΔldhΔals-1 increased lycopene accumulation to 0.70 and 0.73 mg/L, respectively, while their biomasses were reduced by 12.42% and the intracellular NADH/NAD+ ratios increased by 3.05- and 2.10-fold. To increase the biomasses of these engineered strains, aerobic respiration was activated and tuned by the addition of exogenous heme and oxygen. As a result, the engineered L. lactis strains partly recovered the growth and redox balance, yielding the lycopene levels of 0.91-1.09 mg/L. The engineered L. lactis strain protected the intestinal epithelial cells NCM460 against H2O2 challenge, with a 30.09% increase of cell survival and a 29.2% decrease of the intracellular ROS level compared with strain NZ9000 treatment. In summary, this work established the use of the engineered probiotic L. lactis for lycopene production and prospected its potential in the prevention of intestinal oxidative damage.

CEUS Versus MRI in Evaluation of the Effect of Microwave Ablation of Breast Cancer.

Percutaneous microwave ablation (MWA) is a new minimally invasive technique for breast cancer treatment. The aim of this research was to compare the differences in performance between contrast-enhanced ultrasound (CEUS) and magnetic resonance imaging (MRI) with respect to the curative effect of MWA in the treatment of breast cancer. Between 2015 and 2019, 26 patients with breast cancer underwent ultrasound-guided MWA. All patients underwent both CEUS and MRI within 3 d after ablation. If either of the two imaging modalities revealed suspicious enhancement of the ablation zone and the ultrasound-guided biopsy confirmed residual tumor in the suspicious area, supplementary MWA was applied. The diagnostic efficacy of CEUS and MRI within 3 d after ablation was evaluated based on a >6-mo follow-up of 26 patients. Two cases were diagnosed with residual tumors by ultrasound that were missed by MRI. Three cases were diagnosed with residual tumors by MRI that were missed by CEUS. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of CEUS in the diagnosis of complete ablation were 100%, 40%, 87.5%, 100% and 88.5%, respectively. The sensitivity, specificity, PPV, NPV and accuracy of MRI in the diagnosis of complete ablation were 100%, 60%, 91.3%, 100% and 92.3%, respectively. Within 3 d, both CEUS and MRI can efficiently assess the efficacy of MWA of breast cancer.

Risk Factors of Ureteral Stenosis After Percutaneous Microwave Ablation of Renal Tumor, a Single-Center Experience.

BACKGROUND: Ureteral stenosis after percutaneous microwave ablation (MWA) of renal tumor is a rare but severe complication, and its risk factors are not apparent. PURPOSE: This study aimed to investigate the risk factors for stenosis of ureter after MWA treatment of the renal tumor that is a rare complication. MATERIALS AND METHODS: Data of 211 patients who underwent MWA for the treatment of renal tumor were retrospectively analyzed from September 2006 to August 2019. Demographic characteristics, clinical features, ablation parameters, and outcomes were analyzed to find out the potential risk factors of this complication. P < 0.05 is considered significant. RESULTS: Six of 211 patients developed ureter stenosis, and the rate of this complication is 2.84%. The median time of emergence of hydronephrosis was 226 (range, 3-390) days. Univariate analysis shows the distance between ureter and tumor (P = 0.225) or ablation zone (P = 0.089) is not related to this complication. Postoperative urine routine (red blood cell, P = 0.001; white blood cell, P = 0.035) and R.E.N.A.L. score (P < 0.001) is related to this complication. But after multivariate logistic analysis, only R.E.N.A.L. score (P = 0.004) is associated with this complication. The location and growth pattern of tumor and the energy of ablation were not related to this complication independently. CONCLUSION: The stenosis of the ureter after MWA of renal tumor is not associated with the tumor size, location, or the distance between the ureter and tumor and ablation site independently. But R.E.N.A.L. score is associated with ureter stenosis after MWA for the treatment of renal tumor, which combines the information of location, depth, and size of tumor. Preoperative evaluation of the tumor is necessary for avoiding ureter stenosis. Further studies should focus on these risk factors of this complication.

Progesterone decreases gut permeability through upregulating occludin expression in primary human gut tissues and Caco-2 cells.

Progesterone plays a protective role in preventing inflammation and preterm delivery during pregnancy. However, the mechanism involved is unknown. Microbial product translocation from a permeable mucosa is demonstrated as a driver of inflammation. To study the mechanism of the protective role of progesterone during pregnancy, we investigated the effect of physiologic concentrations of progesterone on tight junction protein occludin expression and human gut permeability in vitro and systemic microbial translocation in pregnant women in vivo. Plasma bacterial lipopolysaccharide (LPS), a representative marker of in vivo systemic microbial translocation was measured. We found that plasma LPS levels were significantly decreased during 24 to 28 weeks of gestation compared to 8 to 12 weeks of gestation. Moreover, plasma LPS levels were negatively correlated with plasma progesterone levels but positively correlated with plasma tumor necrosis factor-alpha (TNF-α) levels at 8 to 12 weeks of gestation but not at 24 to 28 weeks of gestation. Progesterone treatment increased intestinal trans-epithelial electrical resistance (TEER) in primary human colon tissues and Caco-2 cells in vitro through upregulating tight junction protein occludin expression. Furthermore, progesterone exhibited an inhibitory effect on nuclear factor kappa B (NF-κB) activation following LPS stimulation in Caco-2 cells. These results reveal a novel mechanism that progesterone may play an important role in decreasing mucosal permeability, systemic microbial translocation, and inflammation during pregnancy.

[Effects of Electroacupuncture and Intracerebral Injection of VEGF on Caspase12, Caspase3, and GRP78 Genes in Rats with Cerebral Ischemia-Reperfusion Injury].

OBJECTIVE: To determine the effects of electroacupuncture (EA) and intracerebral injection of vascular endothelial growth factor (VEGF) on caspase12, caspase3, and glucose regulated protein 78 kD (GRP78) genes of rats with cerebral ischemia reperfusion injury. METHODS: 60 SD rats were randomly divided into sham-operation group, model group, EA group and EA+VEGF group with 15 rats in each group. Middle cerebral artery occlusion (MCAO) method was used to establish the model of cerebral ischemia reperfusion injury. Electro-acupuncture intervention was introduced 1 day after the injury in the EA group and EA+VEGF group: 30 minutes each session and once a day for a total of 14 d [acupoint selection: Baihui (GV 20), Quchi(Li 11), Zusanli (ST36)]. The rats in the EA+VEGF group were also injected with 10 µL of VEGF165 (0.025 µg/µL) into the lateral ventricle after the first session of EA. Five rats in each group were sacrificed after obtaining a neurological function score (mNSS) at day 0 (1 d after modeling, before EA intervention), day 7 and day 14, respectively. Nissl staining was used to observe the histomorphology of cerebral infarction areas. Immunohistochemistry was used to CM(155mm]detect GRP78 activity in the ischemic brain tissues. Real-time fluorescence quantitative PCR (real-time PCR) was used to detect the expressions of caspase12, caspase3 and GRP78 mRNA in the ischemic brain tissues. RESULTS: Compared with the sham-operation group, rats in the model group had higher mNSS scores ( P<0.05), showed signs of cerebral infarction (with reduced numbers of and disordered Nissl bodies and unclear structure), increased GRP78 immunopositive cells, increased expression of GRP78 mRNA ( P<0.05), and increased expressions of caspase12 and caspase3 mRNA ( P<0.05). Compared with the model group, EA and EA+VEGF decreased mNSS scores at day 7 and 14 ( P<0.05), showing alleviated signs of cerebral infarction, increased GRP78 immunopositive cells ( P<0.05), increased GRP78 mRNA expression ( P<0.05), and decreased caspase12 and caspase3 mRNA expressions ( P<0.05). The most obvious changes were found in the EA+VEGF group ( P<0.05). No significant changes were observed in the sham-operation group over time ( P<0.05). In comparison, mNSS scores, the signs of cerebral infarction, and the expressions of caspase12 and caspase3 decreased over time in the other groups ( P<0.05), accompanied with increased GRP78 immunopositive cells and the expression of GRP78 gene ( P<0.05). CONCLUSION: Electroacupuncture and intracerebral injection of VEGF promote tissue repair of rats with cerebral ischemic injury, possibly through down-regulating the expressions of caspase12 and caspase3 genes and up-regulating the expression of GRP78 gene. The effect of electroacupuncture in combination with intracerebral injection of VEGF is superior to that of the single use of electroacupuncture.

[Electroacupuncture Combined with Intracerebral Injection of VEGF Improves Neurological Dysfunction Possibly by Down-regulating Expression of Endoplasmic Reticulum Stress Related Proteins ATF 6, etc. in Cerebral Ischemia-reperfusion Injury Rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) and EA combined with intracerebral injection of vascular endothelial growth factor (VEGF) on endoplasmic reticulum stress (ERS) related proteins and genes as activating transcription factor (ATF 6), inositol requiring enzyme-1 (IRE 1), CCAAT/enhancer binding protein homologous protein (CHOP), X box-binding protein-1 (XBP 1) of cerebral ischemia reperfusion injury (CIRI) rats, so as to study its repair effect for CIRI. METHODS: Forty male SD rats were equally and randomly divided into 5 groups: sham operation, model, EA, VEGF and EA+VEGF groups (n=8). The CIRI model was established by occlusion of the middle cerebral artery (MCAO) with thread embolism method. For rats of the sham operation group, the right common carotid artery was isolated without MCAO. EA (2 Hz/100 Hz, 1-3 mA) was applied to "Baihui" (GV 20), left "Quchi" (LI 11) and left "Zusanli" (ST 36) for 30 min, once a day for 14 days. For rats of the VEGF and EA+VEGF groups, 10 µL VEGF (0.025 µg/µL) was injected into the lateral ventricle 24 h after successful modeling. The rats' neurological function was assessed by using the modified neurological severity score (mNSS), and the histopathological changes of cerebral tissue were observed by Nissl staining method. The expression levels of ERS related proteins and genes ATF 6, IRE 1, XBP 1 and CHOP were determined by western blot (WB) and fluorescent quantitative PCR, separately. RESULTS: After modeling, the level of mNSS was significantly higher in the model group than in the sham operation group (P<0.05), and the number of Nissl bodies was markedly lower in the model group than in the sham operation group (P<0.05). Following the treatment, the mNSS was significantly lower in the EA, VEGF and EA+VEGF groups than in the model group (P<0.05), and the numbers of Nissl bodies were obviously higher in the EA, VEGF and EA+VEGF groups than in the model group (P<0.05), suggesting an improvement of neurological dysfunction and a repair of the injured cerebral tissue after the treatment. The levels of CIRI-induced increase of mNSS and CIRI-induced decrease of the number of Nissl bodies in the EA+VEGF group were respectively remarkably lower or higher than those of the simple EA and simple VEGF groups (P<0.05). WB and PCR showed that the expression levels of ATF 6, IRE 1, XBP1 and CHOP proteins and genes were notably higher in the model group than in the sham operation group (P<0.05), and considerably lower in the EA, VEGF and EA+VEGF groups than in the model group (P<0.05). Comparison among the three treatment groups showed that after the treatment, the expression levels of ATF 6, IRE 1, XBP1 and CHOP proteins and genes were obviously lower in the EA+VEGF group than in the EA and VEGF groups (P<0.05). CONCLUSION: EA and EA plus intracerebral microinjection of VEGF can improve neurological function and promote cerebral tissue repair in CIRI rats, which is associated with their effects in down-regulating the expression of ERS related proteins ATF 6, IRE 1, XBP1 and CHOP. The effect of EA+VEGF is superior to that of simple EA and simple VEGF.

[Electroacupuncture Intervention Improves Physical Power of Aged Rats with Low Testosterone by Reducing Chronic Inflammatory Response of Leydig Cells].

OBJECTIVE: To assess the effect of electroacupuncture (EA) on chronic inflammatory response of Leydig cells in aged rats with low testosterone, so as to investigate its underlying mechanism of anti-male reproductive aging. METHODS: Twenty-four 20 months old SD rats were randomly divided into EA, medication and aged control groups (n＝ 8 in each), and other 8 young SD rats (2 months of age) were used as the youth control group. EA (2 Hz/100 Hz, 1 mA) was applied to "Guanyuan"(CV 4) and bilate-ral "Shenshu"(BL 23) for 15 min, once daily for 8 weeks except the weekends. The medication group received abdominal subcutaneous injection of testosterone propionate (7 mg• kg－1• 3 d－1) for 8 weeks. The aged control group and the youth control group received subcutaneous injection of 0.9% normal saline, with the same dose and same treatment frequency as those of the medication group. The rats' physical power was assessed according to the exhausted swimming duration, and the levels of serum total testosterone (TT) and free testosterone(FT) were determined by ELISA. The pathological changes of the testis tissue were detected by using H．E.staining, and the immunoactivity of cyclooxygenase-2 (COX-2) in Leydig cells was detected by immunohisto-chemistry. The expression levels of nuclear factor-κB p 65 (NF-κB p 65), COX-2, interleukin-1 ß (IL-1 ß) and tumor necrosis factor-α (TNF-α) proteins in the testis tissues were determined by Western blot. RESULTS: Before and after treatment, the exhaustive swimming duration and the levels of serum TT and FT in the aged control group were significantly lower than those of the youth control group (P < 0.01). After the treatment, the exhaustive swimming duration and serum TT and FT in the EA and medication groups were notably higher than those in the aged control group (P<0.01). HE staining showed that the incompleteness of basement membrane of spermatogenic tubules, reduction of spermatogenic cells and supporting cells and irregularity of Leydig cells in the testis tissue of the aged rats were relatively milder after EA intervention. Compared with the youth control group, the expression levels of NF-κB p 65 and COX-2, IL-1 ß and TNF-α in the testicular tissue were significantly higher in the aged control group (P<0.01),while in compared with the aged control group, the expression levels of NF-κB p 65, COX-2, IL-1 ß and TNF-α proteins were significantly down-regulated in the EA group (P<0.01). CONCLUSION: EA intervention can improve the physical power of the aged rats with low testosterone, which may be related to its effects in up-regulating TT and FT levels, and in reducing chronic inflammatory response in the testis tissue.

Flexible Micro-Supercapacitors Based on Naturally Derived Juglone.

Recently, great efforts have been devoted to designing and fabricating flexible, lightweight, wearable, and miniaturized supercapacitors. At the same time, the exploration of green, renewable, and biocompatible energy-storage materials has been attracting intensive attention. By taking fabrication and configuration design into consideration, the naturally derivable juglone molecule was exploited as an active charge-storage material, and integrated into flexible and micro-supercapacitor devices. The polypyrrole/juglone-composite-based supercapacitors exhibit significant energy-storage capabilities with high specific capacitance and long cyclability, which are comparable to that of conventional electrode materials. This study presents a new way for developing flexible, lightweight, portable, and/or wearable electronic devices with biocompatible and environmentally friendly attributes.

Green Synthesis of Red-Emitting Carbon Nanodots as a Novel "Turn-on" Nanothermometer in Living Cells.

Temperature measurements in biology and medical diagnostics, along with sensitive temperature probing of living cells, is of great importance; however, it still faces significant challenges. Herein, a novel "turn-on" carbon-dot-based fluorescent nanothermometry device for spatially resolved temperature measurements in living cells is presented. The carbon nanodots (CNDs) are prepared by a green microwave-assisted method and exhibit red fluorescence (λem =615 nm) with high quantum yields (15 %). Then, an on-off fluorescent probe is prepared for detecting glutathione (GSH) based on aggregation-induced fluorescence quenching. Interestingly, the quenched fluorescence could be recovered by increasing temperature and the CNDs-GSH mixture could behave as an off-on fluorescent probe for temperature. Thus, red-emitting CNDs can be utilized for "turn-on" fluorescent nanothermometry through the fluorescence quenching and recovery processes, respectively. We employ MC3T3-E1 cells as an example model to demonstrate the red-emitting CNDs can function as "non-contact" tools for the accurate measurement of temperature and its gradient inside a living cell.