In vivo evaluation of liver function by multimodal imaging in an alcohol-induced liver injury model.

Background: Visually evaluating liver function is a hot topic in hepatology research. There are few reliable and practical visualization methods for evaluating the liver function in vivo in experimental studies. In this study, we established a multimodal imaging approach for in vivo liver function evaluation and compared healthy mice with chronic alcoholic liver injury (cALI) model mice to explore its potential applicability in experimental research. Methods: In vivo fluorescence imaging (IVFI) technology was utilized to visually represent the clearance of indocyanine green from the liver of both healthy mice and mice with cALI. The reserve liver function was evaluated via IVFI using the Cy5.5-galactosylated polylysine probe, which targets the asialoglycoprotein receptor of hepatocytes. Hepatic microcirculation was assessed through laser speckle perfusion imaging of hepatic blood perfusion. The liver microstructure was then investigated by in vivo confocal laser endomicroscopy imaging. Finally, hepatic asialoglycoprotein receptor expression, histology, and the levels of serum alanine aminotransferase and aspartate aminotransferase were measured. Results: In vivo multimodal imaging results intuitively and dynamically showed that indocyanine green clearance [mean ± standard deviation (SD): 30.83±14.71, 95% confidence interval (CI): 20.3 to 41.35], the fluorescence signal intensity (mean ± SD: 1,217.92±117.63; 95% CI: 1,148.38 to 1,290.84) and fluorescence aggregation area (mean ± SD: 5,855.80±1,271.81; 95% CI: 5,051.57 to 6,653.88) of Cy5.5-galactosylated polylysine targeting the asialoglycoprotein receptor, and hepatic blood perfusion (mean ± SD: 1,494.86±299.33; 95% CI: 1,316.98 to 1,690.16) in model mice were significantly lower than those in healthy mice (all P<0.001). Compared to healthy mice, the model mice exhibited a significant decline in liver asialoglycoprotein receptor expression (mean ± SD: 219.03±16.34; 95% CI: 208.97 to 230.69; P<0.001), increased serum alanine aminotransferase (mean ± SD: 149.70±47.89 U/L; 95% CI: 81.75 to 128.89; P=0.01) and aspartate aminotransferase levels (mean ± SD: 106.30±36.13 U/L; 95% CI: 122.01 to 180.17; P=0.021), hepatocyte swelling and deformation, disappearance of the hepatic cord structure, partial necrosis, and disintegration of hepatocytes. The imaging features of fluorescence signals in liver regions, hepatic blood perfusion and microstructure were biologically related to hepatic asialoglycoprotein receptor expression, serum indices of liver function, and histopathology in model mice. Conclusions: Utilizing in vivo multimodal imaging technology to assess liver function is a viable approach for experimental research, providing dynamic and intuitive visual evaluations in a rapid manner.

Association between urinary neonicotinoid insecticide levels and dyslipidemia risk: A cross-sectional study in Chinese community-dwelling elderly.

Experimental evidence has demonstrated that neonicotinoids (NEOs) exposure can cause lipid accumulation and increased leptin levels. However, the relationship between NEOs exposure and dyslipidemia in humans remains unclear, and the interactive effects of NEOs and their characteristic metabolites on dyslipidemia remain unknown. We detected 14 NEOs and their metabolites in urine samples of 500 individuals (236 and 264 with and without dyslipidemia, respectively) randomly selected from the baseline of the Yinchuan community-dwelling elderly cohort (Ningxia, China). The NEOs and their metabolites were widely detected in urine (87.2-99.6 %) samples, and the median levels ranged within 0.06-0.55 µg/g creatinine. The positive associations and dose-dependent relationships of thiacloprid, imidacloprid-olefin, and imidacloprid-equivalent total with dyslipidemia were validated using restricted cubic spline analysis. Mixture models revealed a positive association between the NEOs mixture and dyslipidemia risk, with urine desnitro-imidacloprid ranked as the top contributor. The Bayesian Kernel Machine Regression models showed that the NEOs mixtures were associated with increased dyslipidemia when the chemical mixtures were ≥ 25th percentile compared to their medians, and desnitro-imidacloprid and imidacloprid-olefin were the major contributors to the combined effect. Given the widespread use of NEOs and the dyslipidemia pandemic, further investigations are urgently needed to confirm our findings and elucidate the underlying mechanisms.

Berberine regulates glucose metabolism in largemouth bass by modulating intestinal microbiota.

This study examined the role of intestinal microbiota in berberine (BBR)-mediated glucose (GLU) metabolism regulation in largemouth bass. Four groups of largemouth bass (133.7 ± 1.43 g) were fed with control diet, BBR (1 g/kg feed) supplemented diet, antibiotic (ATB, 0.9 g/kg feed) supplemented diet and BBR + ATB (1g/kg feed +0.9 g/kg feed) supplemented diet for 50 days. BBR improved growth, decreased the hepatosomatic and visceral weight indices, significantly downregulated the serum total cholesterol and GLU levels, and significantly upregulated the serum total bile acid (TBA) levels. The hepatic hexokinase, pyruvate kinase, GLU-6-phosphatase and glutamic oxalacetic transaminase activities in the largemouth bass were significantly upregulated when compared with those in the control group. The ATB group exhibited significantly decreased final bodyweight, weight gain, specific growth rates and serum TBA levels, and significantly increased hepatosomatic and viscera weight indices, hepatic phosphoenolpyruvate carboxykinase, phosphofructokinase, and pyruvate carboxylase activities, and serum GLU levels. Meanwhile, the BBR + ATB group exhibited significantly decreased final weight, weight gain and specific growth rates, and TBA levels and significantly increased hepatosomatic and viscera weight indices and GLU levels. High-throughput sequencing revealed that compared with those in the control group, the Chao one index and Bacteroidota contents were significantly upregulated and the Firmicutes contents were downregulated in the BBR group. Additionally, the Shannon and Simpson indices and Bacteroidota levels were significantly downregulated, whereas the Firmicutes levels were significantly upregulated in ATB and BBR + ATB groups. The results of in-vitro culture of intestinal microbiota revealed that BBR significantly increased the number of culturable bacteria. The characteristic bacterium in the BBR group was Enterobacter cloacae. Biochemical identification analysis revealed that E. cloacae metabolizes carbohydrates. The size and degree of vacuolation of the hepatocytes in the control, ATB, and ATB + BBR groups were higher than those in the BBR group. Additionally, BBR decreased the number of nuclei at the edges and the distribution of lipids in the liver tissue. Collectively, BBR reduced the blood GLU level and improved GLU metabolism in largemouth bass. Comparative analysis of experiments with ATB and BBR supplementation revealed that BBR regulated GLU metabolism in largemouth bass by modulating intestinal microbiota.

Effects of electroacupuncture on liver function in mice with chronic alcoholic liver injury: visual display by in vivo fluorescence imaging.

OBJECTIVE: Acupuncture can improve the symptoms of alcohol-induced bodily injury and has been accepted by the World Health Organization. In this study, in vivo fluorescence imaging (IVFI) was applied to display and evaluate the effect of electroacupuncture (EA) on liver function (LF) in mice with chronic alcoholic liver injury (cALI). METHODS: IVFI of the Cy5.5-galactosylated polylysine (Cy5.5-GP) probe targeting the liver asialoglycoprotein receptor (ASGPR) and liver indocyanine green (ICG) clearance was performed to visually evaluate the effect of EA at ST36 and BL18 on liver reserve function and hepatic metabolism in mice with cALI. In addition, changes in ASGPR expression, serum indexes of LF, and hepatic morphology were observed. RESULTS: After EA at ST36 and BL18, the ASGPR-targeted fluorescence signals (FS) in the liver increased significantly in cALI mice (p < 0.05) and exhibited relationships with liver ASGPR expression, liver ICG clearance, liver histology, and serum marker levels of LF in cALI mice undergoing EA intervention. CONCLUSIONS: As displayed by IVFI, EA at ST36 and BL18 appears to improve liver reserve function and inhibit the development of liver injury in mice with cALI. EA may have potential as a treatment strategy to protect against ALI.

A Novel Cuproptosis-Related Signature Identified DLAT as a Prognostic Biomarker for Hepatocellular Carcinoma Patients.

Background: Hepatocellular carcinoma (HCC) is the most common type of liver cancers, with more than a million cases per year by 2025. Cuproptosis is a novel form of programmed cell death, and is caused by mitochondrial lipoylation and destabilization of iron-sulfur proteins triggered by copper, which was considered as a key player in various biological processes. However, the roles of cuproptosis-related genes (CRGs) in HCC remain largely unknown. Methods: In the present study, we constructed and validated a four CRGs signature for predicting the overall survival (OS) of HCC patients in both The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) databases. Results: Patients with high CRGs risk score showed shorter OS than those with low CRGs risk score. Functional analysis suggested that the CRGs-based prognostic signature was associated with metabolism remodeling which facilitated liver cancer progression. In addition, reduced infiltration of CD8+ T cells and increased macrophages were found in HCCs from patients with high CRGs risk score. As one of the four CRGs, higher expression of dihydrolipoamide S-acetyltransferase (DLAT) was accompanied by higher expression of program death ligand 1 (PD-L1) in HCC. Further, we confirmed that DLAT was up-regulated and correlated with poor prognosis in a clinical HCC cohort. Conclusion: In conclusion, our study constructed a four CRGs signature prognostic model and identified DLAT as an independent prognostic factor for HCC, thus providing new clues for understanding the association between cuproptosis and HCC.

New Insights into the Relationships between Bacterial Dynamics and Water Quality of Aquaculture Systems Supplemented with Carbon Source and Biofilm Substratum.

Aquaculture is crucial for achieving the FAO's goal of a world without hunger and malnutrition. Recently, biofilm substratum has been proposed as an effective means to control waste pollution caused by excessive nutrient inputs from aquaculture, but key bacterial communities involved in the remediation remain unclear. Here we reported a freshwater mesocosm study where the addition of biofilm substrata with external carbon effectively controlled the total ammonia nitrogen and improved fish growth. 16S rRNA study and Weighted UniFrac analysis revealed that bacterial compositions were significantly different (999 permutations, p-value < 0.01) between the biofilm-substrata-added and biofilm-substrata-free systems. Planctomycetes were found, as key bacteria benefited from the biofilm substrata addition and exerted the major function of ammonia nitrogen control. Our study demonstrated that the addition of biofilm substrata and an external carbon source favored fish growth and improved the aquaculture environment by the formation of a unique bacteria community.

[Effect of acupuncture on the cognitive function of migraine patients with depression/anxiety disorder].

OBJECTIVE: To observe the effect of five-element acupuncture on the cognitive function repair of migraine patients with depression/anxiety disorder. METHODS: The migraine patients with depression/anxiety disorder (19 cases, 5 cases dropped off) were taken as the observation group, and received five-element acupuncture twice a week for 8 weeks. Healthy subjects (19 cases) were selected by demographic data matching as the control group. The cognitive function was evaluated with the event related potential (ERP) technique, and the latency and amplitude of visual evoked potential P300 were adopted as the observation indexes. The headache days (every 4 weeks), headache intensity [visual analogue scale(VAS) score], and headache impact test-6 (HIT-6) score, Hamilton depression scale (HAMD) score and Hamilton anxiety scale (HAMA) score were used as the observation indexes for curative effect. RESULTS: Before the treatment, latency of target stimulus at Fz [ (417.5±34.3) ms] in the observation group was extended compared with the healthy subjects of the control group [(388.6±42.1) ms, P<0.05]. In the observation group, the latency of each point target stimulus [Fz: (376.1±36.2) ms, F3: (374.8±37.6) ms, F4: (372.0±37.6) ms] after treatment were shorter than those [Fz: (417.5±34.3) ms, F3: (417.4±33.8) ms, F4: (416.0±36.6) ms] before treatment (P<0.05). Before and after treatment, there was no significant difference in the amplitude of each point between the observation group and the control group (P>0.05). In the observation group, the headache days was shorter than that before treatment (P<0.01), and the VAS score, HIT-6 score, HAMD score and HAMA score were all lower than before treatment (P<0.01). CONCLUSION: There are some cognitive impairments in migraine patients with depression/anxiety disorder. Five-element acupuncture not only relieves headache, anxiety and depression effectively, but also improves the activation level of the frontal lobe. It significantly repairs the impaired cognitive function.

[Sixty years of meridian-collateral exploration: a road of interdisciplinary research].

The theory of traditional Chinese medicine believes that meridian-collateral (jingluo) is an important system to maintain life and participates in the formation and cure of disease. Exploring the biological connotation of meridian-collateral is the key content in the modern research of traditional Chinese meridian. For more than 60 years, scholars at home and abroad have carried out the researches of meridian-collateral from the fields of biomedicine (e.g. physiology, anatomy and histomorphology), imaging, physics, chemistry, computer technology, etc. Meridian-collateral research in modern times demonstrates a multi-disciplinary historical development and provides multi-perspective scientific information for understanding meridian-collateral. The meridian-collateral research in future needs more accurate multidisciplinary intersection, which will be the light of hope to reveal the biological connotation of meridian-collateral.

[Effect of electroacupuncture on the hemorheology and hepatic microcirculation in mice with chronic alcoholic liver injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the hemorheology and hepatic microcirculation in mice with chronic alcoholic liver injury (cALI), so as to explore the microcirculation mechanism of EA underlying regulating liver function in cALI mice. METHODS: Forty Kunming mice were randomly assigned to control, model, acupoint EA and non-acupoint EA groups, with 10 mice in each group. The cALI model was established by gavage with 50% ethanol (15 mL/kg，2 times per day with 8 hours' interval) for 28 d. Mice in the acupoint EA group received EA at bilateral "Zusanli" (ST36) and "Ganshu"(GB11) for 20 min, once daily for 14 d. And mice in the non-acupoint EA group received EA the spot about 1 cm lateral to GB11 and 0.5 cm lateral to ST36 respectively for 20 min, once daily for 14 d. The hepatic blood perfusion (HBP) of mice was detected by laser speckle perfusion imaging. The index of hemorheology, serum markers of liver function and hepatic histology in mice were observed by automatic blood rheometer, automatic bio-analysis machine and H.E. staining, respectively. RESULTS: Compared with the control group, the HBP of the model group was significantly decreased (P<0.05); the low-cut whole blood viscosity, plasma viscosity, and erythrocyte sedimentation level were significantly increased （P<0.05）, and the red blood cell deformation index was significantly decreased (P<0.05); serum alanine aminotransferase (ALT) and gamma-glutamyl transferase (GGT) increased significantly (P<0.05) in the model group. Hepatocyte steatosis accompanied by inflammatory cell infiltration and focal necrosis was observed in the model group. After EA at ST36 and GB11, and in comparison with the model group showed that the HBP was significantly increased (P<0.05); the low-cut whole blood viscosity, plasma viscosity, erythrocyte sedimentation rate and serum GGT were significantly decreased (P<0.05), and the red blood cell deformation index was significantly increased (P<0.05）; the degree of fatty degeneration in the liver tissue was reduced, and the focal necrosis was reduced. The plasma viscosity, erythrocyte sedimentation rate were significantly decreased (P<0.05), and the red blood cell deformation index was significantly increased in the non-acupoint EA group relevant to the model group (P<0.05). The erythrocyte sedimentation rate and the red blood cell deformation index of the acupoint EA group was significantly lower than that of the non-acupoint EA group (P<0.05). CONCLUSION: EA at ST36 and GB11 could improve the hemorheological disorder and the hepatic microcirculation, and inhibit the deve-lopment of liver injury in cALI mice.

Curcumin alleviates acute kidney injury in a dry-heat environment by reducing oxidative stress and inflammation in a rat model.

Curcumin exhibits anti-inflammatory and antioxidant activities. We investigated the protective effects of curcumin in a renal injury rat model under dry-heat conditions. We divided Sprague-Dawley rats into four groups: dry-heat 0- (normal temperature control group), 50-, 100-, and 150-minute groups. Each group was divided into five subgroups (n = 10): normal saline (NS), sodium carboxymethylcellulose (CMCNa), and curcumin pretreated low, medium, and high-dose (50, 100, and 200 mg/kg, respectively) groups. Compared to the normal temperature group, serum creatinine, blood urea nitrogen, urinary kidney injury molecule-1, and neutrophil gelatinase-associated load changes in lipoprotein (NGAL) levels were significantly increased in the dry-heat environment group (P < .05); inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression and malondialdehyde (MDA) and related inflammatory factor levels were increased in the kidney tissue. Superoxide dismutase (SOD) and catalase (CAT) levels were decreased. However, following all curcumin pretreatment, the serum levels of kidney injury indicators and NGAL were decreased in the urine compared to those in the NS and CMCNa groups (P < .05), whereas renal SOD and CAT activities were increased and MDA was decreased (P < .05). Renal tissues of the 150-minute group showed obvious pathological changes. Compared to the NS group, pathological changes in the renal tissues of the 100- and 200-mg/kg curcumin groups were significantly reduced. Furthermore, iNOS and COX-2 expression and inflammatory factor levels were decreased after curcumin treatment. Curcumin exerted renoprotective effects that were likely mediated by its antioxidant and anti-inflammatory effects in a dry-heat environment rat model.

Identification and analysis of lipid droplet-related proteome in the adipose tissue of grass carp (Ctenopharyngodon idella) under fed and starved conditions.

Fat accumulation in the mesenteric adipose tissue is a serious problem in grass carp (Ctenopharyngodon idella) culture. Lipid droplet-related proteins (LDRPs) are involved in the formation, degradation, and biological functions of lipid droplets. In this study, we aimed to provide reference proteomics data to study lipid droplet regulation in fish. We isolated LDRPs from the mesenteric adipose tissue of grass carp (1-year-old) after normal feeding and 7 days of starvation, and identified and analysed them using isobaric tags for relative and absolute quantitation (iTRAQ) technology. Short-term starvation had no significant effect on the body weight, condition factor, visceral index, hepatopancreas index, intraperitoneal fat index, adipose tissue triglyceride content, and adipocyte size of grass carp. Nine hundred and fifty proteins were identified and annotated using the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases; they are involved in a variety of metabolic and signalling pathways, including amino acid, lipid, and carbohydrate metabolism, and the PI3K-Akt signalling pathway. There were 296 differentially expressed proteins (DEPs), with 143 up-regulated and 153 down-regulated proteins. Three proteins involved in triglyceride and fatty acid syntheses and two proteins involved in autophagy were up-regulated, and six proteins involved in lipid catabolism were down-regulated. These results indicate that under short-term starvation, lipid droplets in the adipose tissue of grass carp may maintain their shape by promoting fat production and inhibiting lipolysis, and autophagy may be one of the main strategies for coping with short-term energy deprivation.

[Consumption Capacity of N2O in Paddy Soil and the Response Mechanism of nosZ-I-containing Communities].

The long-term flooding anaerobic environment in paddy soils is conducive to denitrification, which is one of the most important reasons for N2O emissions. N2O can be transformed to nitrogen gas (N2) by bacteria and archaea containing nitrous oxide reductase (N2OR) encoded by the nosZ gene, which is the only known biological pathway of N2O consumption in soil. nosZ-I is known to be typical in denitrifying bacteria, which is one of the clades of the nosZ gene and is mainly possessed a Tat signal peptide motif. Although many researchers have studied N2O emission characteristics of paddy soil, the capacity of N2O consumption and the response mechanism of related functional microorganisms in paddy fields is not yet clear. To verify the effect of exogenous N2O on N2O consumption and nosZ-I gene, a pot trial experiment was performed under anaerobic conditions. We collected intact soil cores from flooding paddy fields at a 0-5 cm depth, and exogenous N2O gas was input through the bottom of flooding paddy soil cores. Meanwhile, a control treatment (CK) with no additional N2O gas was also performed. The dynamic characteristics of the added exogenous N2O concentration through the intact soil cores, the content of inorganic nitrogen, and DOC were systematically monitored. In addition, the change in the nosZ-I population diversity and community composition were investigated by high-throughput sequencing approaches, with the purpose of revealing the N2O uptake ability of flooded paddy soil and the response mechanism of the nosZ-I population. The results showed that 97.39% of exogenous N2O diffused into the soil cores, and only 0.72%-7.75% of exogenous N2O escaped from the soil surface. The N2O released in the headspace of soil cores could continue being absorbed and consumed by the flooding soil column. In addition, 67.10% of the N2O escaped to the headspace was consumed in exogenous N2O treatment after 192 h of incubation, which was higher than that in CK treatment, and the N2O consumption rate increased by 144.2% than that in CK treatment. Meanwhile, the consumption of NH4+-N, NO3--N, and DOC consumed during exogenous N2O addition treatment was 19.65%, 16.29%, and 8.41% higher than that in CK treatment, respectively. However, the diversity of the nosZ-I gene community had no significant difference; the community composition of nosZ-I-containing bacteria changed significantly after 192 h when exogenous N2O was input. The abundances of OTU5004, OTU5065, OTU960, and OTU1282 (Proteobacteria) significantly increased, which were the dominant bacterial strain of nosZ-I gene on the OTU level. Compared with the initial sample and CK, the abundance of the OTU5004 strain increased by 7.3% and 4.63%, and the abundance of the OTU5265 strain (Azoarcus sp.) increased by 0.33% and 0.15%, respectively. The result indicated that the flooding paddy soil column at the soil layer of 0-5 cm has a strong N2O absorption and consumption ability. In summary, compared with CK, the addition of exogenous N2O significantly accelerated the N2O consumption rate, improved the consumption potential of flooding paddy soil column, promoted carbon and nitrogen conversion, and changed nosZ-I community composition. These results would provide a new reference for reducing atmospheric N2O emissions.

Feeding Faba Beans (Vicia faba L.) Reduces Myocyte Metabolic Activity in Grass Carp (Ctenopharyngodon idellus).

In this study, we aimed to explore the effects of faba bean (Vicia faba L.) on the energy metabolism of grass carp (Ctenopharyngodon idellus). A total of 180 fish (∼2900 g) were randomly assigned to six tanks (2.5 × 2.5 × 1.2 m; 30 individuals per tank) and fed either faba bean (Vicia faba L.) or a commercial diet for 120 days (3% body weight, twice per day). The results showed that faba bean-fed grass carp (FBFG) had significantly lower growth and higher fat accumulation in the mesenteric adipose tissue and hepatopancreas than commercial diet-fed grass carp (CDFG). Compared with CDFG, FBFG exhibited no significant difference in proximate composition of the muscle; however, an obvious decrease in muscle fiber size and significantly higher hardness, chewiness, and gumminess were observed. Transcriptome results showed that a total of 197 genes were differentially regulated in the dorsal muscle. Down-regulated genes included four genes annotated with myocyte development and 12 transcripts annotated with components of myofibrils. In addition, the FBFG group exhibited significantly lower expression of genes associated with oxygen transport, the mitochondrial respiratory chain, and creatine metabolism, suggesting reduced energy availability in the muscle of the FBFG. Moreover, using western-blotting and enzyme assays, we found decreased protein levels in the mitochondrial electron transport respiratory chain and creatine metabolism activities, as well as increased expression of autophagy marker protein levels, in the muscle of FBFG. Overall, our results suggest that an abnormal energy distribution may exist in grass carps after feeding with faba bean, which is reflected by a mass of fat deposition in the adipose tissue and hepatopancreas and subdued metabolic activity in the muscle.

[Display of low hydraulic resistance channels along Conception Vessel by in vivo fluorography in rats].

OBJECTIVE: To display the low hydraulic resistance channel along the running course of the Conception Vessel (CV) by using in vivo fluorography, so as to provide evidence for the existence of meridians of traditional Chinese medicine from the aspect of interstitial fluid channel. METHODS: A total of 24 male SD rats were randomly divided into 3 groups, namely CV-low impedance spots (CV-LIS), CV-low hydraulic resistance points (CV-LHRP) and non-meridian points (NMP) groups (n＝8 in each group). The CV-LIS (the imaged line between the xiphoid process and symphysis pubis), CV-LHRP (lateral and parallel to LIS) and NMP (lateral and parallel to CV-LHRP) were detected by using meridian detector and biofluid resistance meter, respectively. Sodium fluorescein solution (1%) was injected into the spot (A-spot, upper 1/5 of the CV-LIS), and one of the NMP (about 1 cm lateral and parallel to the A-spot, and the mid-point between the Kidney and Stomach meridian running courses) by using a microsyringe for observing the migration trace of sodium fluorescein. RESULTS: After injection of fluorescein, the migration trace of fluorescein appeared along the CV-LIS and CV-LHRP lines, and only local diffusion was found in the NMP group. The migration trace lengths along the CV-LIS and CV-LHRP were significantly longer than that of the NMP group (P<0.01), and the migration width of CV-LIS and CV-LHRP groups was significantly smaller than that of the NMP group (P<0.01). The migration speed of CV-LIS and CV-LHRP groups was significantly higher than that of the NMP group (P<0.01). CONCLUSION: Fluorography is able to be used to display the running course of CV, possibly supporting the existence of meridians from the interstitial fluid channel and providing a new method for visualization of meridians.

[Preliminary observation on imbalance degree of impedance in extracellular fluid of cells in the twelve meridians in healthy subjects].

OBJECTIVE: To calculate the imbalance degree (IBD) of left-right meridian (IBD-LRM), IBD of exterior-interior meridian (IBD-EIM) and IBD of hand-foot meridians (IBD-HFM) of impedance in extracellular fluid of cells in twelve meridians of healthy subjects, so as to provide foundation for meridian diagnosis. METHODS: A total of 31 healthy volunteers were enrolled and bioelectrical impedance spectroscopy (BIS) was applied. The constant current (from 1 to 100 kHz, 200 µA) was connected into the bilateral twelve meridians through two excitation electrodes with a distance of 10 cm. Two measuring electrodes, with an interval of 5 cm, were set in between the two excitation electrodes to collect the voltage amplitude and phase. The Cole-Cole curve fitting was used to calculate the impedance of extracellular fluid of cells in the twelve meridians; the IBD-LRM, IBD-EIM and IBD-HFM as well as their absolute values were calculated. RESULTS: The impedance of extracellular fluid in the left side was higher than that in right side in the large intestine meridian, the small intestine meridian and the bladder meridian (P<0.05, P<0.01). The mean value of IBD-LRM of extracellular fluid was (4.0±1.4) %; the mean value of absolute value of IBD-LRM was (15.0±1.1) %; the maximum absolute value of IBD-LRM was the bladder meridian. The mean value of IBD-EIM was (3.3±1.0) %; the mean value of absolute value of IBD-EIM was (17.9±1.6) %; the maximum absolute value of IBD-EIM was the bladder meridian and the kidney meridian. The impedance of extracellular fluid of hand jueyin meridian, hand taiyin meridian and hand shaoyin meridian were lower than those of foot meridians. The mean value of IBD-HFM was (-2.6±1.1) %; the mean value of absolute value of IBD-HFM was (19.7±1.7) %; the maximum absolute value of IBD-HFM was shaoyang meridian; the imbalance of yin meridians was greater than yang meridians. There were significant differences in impedance of extracellular fluid between left and right and between hands and feet (P<0.05, P<0.01). CONCLUSION: The extracellular fluid of left-right meridians of healthy subjects is different, but the absolute value of IBD is low; the mean value of exterior meridian and interior meridian is very close, and the absolute value of IBD is medium; the impedance of the foot meridians are greater than the hand meridians, and the absolute value of IBD is relatively high.

[Impact of Dicyandiamide (DCD) and 3,4-Dimethylpyrazole Phosphate (DMPP) on Ammonia-oxidizing Bacteria and Archaea in a Vegetable Planting Soil].

Nitrification inhibitors (NIs) dicyandiamide (DCD) and 3,4-dimethylpyrazole phosphate (DMPP) showed significant effects in the inhibition of nitrification and the improvement of the utilization efficiency of nitrogen fertilizer in agricultural soils. However, the effects of different NIs on ammonia-oxidizing bacteria (AOB) and archaea (AOA) is still unclear. To verify the inhibitory effect of DCD and DMPP on AOB and AOA, a pot experiment was performed, including Urea, Urea+DCD, and Urea+DMPP treatments. The dynamics of NH4+-N and NO3--N and nitrification potential among different treatments were measured. In addition, real-time PCR and high-throughput sequencing approaches were applied to investigate the changes in the AOB and AOA population abundance and composition. The results revealed that the concentrations of NH4+-N in Urea+DCD and Urea+DMPP treatments were 213% and 675% higher than that in the CK treatment, respectively. However, the concentrations of NO3--N and the nitrification potentials were 13.3% and 37.2%, and 20.4% and 82.4% lower than that in CK treatment, respectively; Furthermore, the copy numbers of the bacterial and archaeal amoA gene were 51.2% and 56.5%, and 6.0% and 27.0% lower than that in the CK treatment, respectively. However, the diversity indexes of AOB and AOA communities, including evenness and richness, exhibited no significant differences after addition of DCD and DMPP. The nork-environmental-samples, unclassified-Nitrosomonadaceae, unclassified-Bacteria, and Nitrosospira, were the predominant genera of the AOB community. The no rank-Crenarchaeota, no rank-environmental-samples and Nitrososphaera were the predominant groups in the AOA community. Summarily, application of DCD and DMPP significantly delayed the transformation of NH4+-N, decreased the formation of NO3--N, inhibited the abundance and changed the composition of AOB and AOA communities. DMPP had a stronger inhibitory effect on nitrification, and on AOB and AOA than DCD. Therefore, compared with DCD, DMPP had a better application prospect regarding the improvement of the nitrogen utilization efficiency in vegetable soil.

Lipid droplets participate in modulating innate immune genes in Ctenopharyngodon idella kidney cells.

Lipid droplets (LDs) are increasingly being recognized as important immune modulators in mammals, in additional to their function of lipid ester deposition. However, the role of LDs in fish immunity remains poorly understood. In this study, the function of LDs in the innate immune response of Ctenopharyngodon idella kidney (CIK) cells, which are the equivalent of myeloid cells in vertebrates, was investigated. LD number and TG content significantly increased in the CIK cells following exposure to lipopolysaccharide (LPS), peptidoglycan (PGN), and polyriboinosinic-polyribocytidylic acid (Poly [I: C]) for 24 h, accompanied by increases in the relative expression of several innate immune genes. However, fatty acid compositions of the triglycerides were not changed after treatment with these three pathogenic mimics. LPS, PGN, and Poly (I: C) did not alter the relative expressions of lipogenic (FAS, SCD, and DGAT) and lipid catabolic (PPARα, ATGL, and CPT-1) genes. However, these treatments did increase the mRNA levels of lipid transportation genes (FATP/CD36, ACSL1, and ACSL4), and also decreased the non-esterified fatty acid level in the medium. To further explore the role of LDs in the immune response, CIK cells were incubated with different concentrations (0, 100, 200, 300, 400, 500 µM) of exogenous lipid mix (LM; oleic acid [OA]:linoleic acid [LA]:linolenic acid [LNA] = 2:1:1), and were then transferred to a lipid-free medium and incubated for 24 h. LD size and number increased with the increase in lipid levels, and this was accompanied by increased expression of innate immune genes, including MyD88, IRF3, and IL-1ß, which were expressed at their highest levels in 300 µM exogenous lipid mix. Interestingly, after incubating with different fatty acids (LM, OA, LA, LNA, arachidonic acid [ARA], and docosahexaenoic acid [DHA]; 300 µM), ARA and DHA were more potent in inducing LD formation and innate immune gene expression in the CIK cells. Finally, atglistatin, an ATGL inhibitor, effectively attenuated the expression of most genes upregulated by ARA or DHA, suggesting that lipolysis may be involved in the regulation of immune genes at the transcriptional level. Overall, the findings of this study demonstrate that LDs are functional organelles that could act as modulators in the innate immune response of CIK cells. Additionally, long-chain polyunsaturated fatty acid enriched LDs play a unique role in regulating this process.

Smad4-dependent regulation of type I collagen expression in the muscle of grass carp fed with faba bean.

Smad4 is the key regulator in the transforming growth factor ß1 (TGF-ß1)/Smads signal pathway, and is also the crux of the regulation of type I collagen expression in mammals. In fish, however, the relationship between Smad4 and type I collagen is still unknown. Given the widely accepted importance of type I collagen in fish muscle hardness, we seek to explore this issue by analyzing the expressions of the TGF-ß1/Smads pathway molecules and type I collagen in the muscle of crisp grass carp fed with faba bean, which shows increased muscle hardness. The study found that (1) in the process of feeding the grass carp with faba bean, the mRNA and protein expressions of TGF-ß1, Smad2 and Smad4 all increased along with the increase of type I collagen expression (Col1α1 and Col1α2); (2) one day after the injection of Smad4 over-expression vector, both mRNA and protein expressions of Col1α1 and Col1α2 significantly increased, reaching the maximum on the 2nd and 5th day, respectively; (3) one day after the injection of Smad4 RNAi interference vector, the mRNA and protein expressions of Col1α1 and Col1α2 decreased, reaching the minimum on the 5th day. These results revealed that Smad4 is the major regulator of type I collagen in the muscle of grass carp fed with faba bean. This study would provide an important mechanistic basis for nutritional regulation of type I collagen in the muscle of fish.

Lipid accumulation in grass carp (Ctenopharyngodon idellus) fed faba beans (Vicia faba L.).

Feeding faba beans (Vicia faba L.) to grass carp (Ctenopharyngodon idellus) increases muscle compactness but decreases growth and motility. The lipid metabolism of grass carp was examined to assess potential effects of feeding faba beans on physiological properties using a total of 180 fish. The treatment group was fed faba beans for 120 days and a commercial diet for another 30 days. The control group received a commercial diet for 150 days. Fish were sampled every month. Weight gain was significantly lower in the treatment group than in the control. Hardness, springiness, chewiness, cohesiveness, and gumminess of the dorsal muscle increased significantly with the feeding faba beans from 30 to 120 days, which was not reversed by the subsequent feeding of commercial diet. Fat accumulation increased significantly in the treatment group as suggested by the condition factor, viscera index, hepatopancreatic index, and intraperitoneal fat index (IPFI), hepatopancreas, and muscle fat content but was not affected by subsequent feeding with the commercial diet. Serum triglyceride and total cholesterol levels were significantly reduced in the experimental diet group. In the hepatopancreas and intraperitoneal fat IPF, monounsaturated fatty acids showed significantly higher content in faba bean feeding fish, whereas polyunsaturated fatty acid content showed the reversed pattern. In the hepatopancreas, the activities of the lipogenic enzymes malate dehydrogenase and glucose 6-phosphate dehydrogenase were higher in the treatment than in the control group. Moreover, the treatment group showed lower mRNA levels of carnitine palmitoyltransferase-1. Overall, our results clearly demonstrate increasing lipid accumulation in the viscera of faba bean-fed grass carp.

[Comparison of Extracellular and Intracellular Fluid Distribution Between Stomach Meridian or Gallbladder Meridian and Their Neighboring Tissues of Lower Leg in Healthy Volunteers].

OBJECTIVE: To compare differences of extracellular fluid impedance (Re) and intracellular fluid impedance (Ri) between the Stomach(ST) Meridian or Gallbladder(GB) Meridian and their neighboring non-meridian sites of the left lower leg at the same level, so as to explore the distribution characteristics of body fluid in the meridian. METHODS: Sixteen healthy volunteers were enrolled in the present study. The Re and Ri were detected by using Ag/AgCl electrodes and a digital lock-in amplifier. The measuring electrodes (at an interval of about 3 cm) were separately fixed to the skin sites covering the running courses of the ST Meridian (in the lateral interspace of the anterior tibial muscle)and the GB Meridian (in the interspace of the anterior edge of the fibula), and the excitation electrodes (at an interval of about 9 cm) respectively fixed to the skin sites covering the anterior tibial muscle and the interspace between the anterior tibial muscle and the tibia (about 2 cm and 5 cm lateral to the ST and GB meridians, and about 3－4 cm and 6－8 cm lateral to the ST and GB meridians, respectively). A 100 µA constant current with frequencies from 1 kHz to 100 kHz delivered via an excitation electrode was applied to the site (control spots of the ST Meridian), and signals of the voltage amplitude and phase difference of the tissues fed to the lock-in amplifier via the measuring electrode were collected, followed by measuring those of the GB Meridian and control sites. The circumference of the lower leg around the two excitation and measuring electrodes was measured. Then the cole-cole curve fitting was performed to calculate the Ri and Re, as well as the intracellular fluid resistivity (ρi) and extracellular fluid resistivity (ρe) of the ST and GB meridians, the related muscles and interspace lateral to ST or GB (ST/GB) meridians at the same level. RESULTS: The Ri and Re (Ω) values of the ST, GB, the muscle lateral to ST/GB and the interspace lateral to ST/GB were 19.1±1.3 and 28.3±1.4, 15.8±1.9 and 25.7±2.0, 19.6±1.3 and 31.3±1.6, and 19.4±1.2 and 32.4±1.6, respectively. The Re values were significantly lower at the ST and GB meridians than at the muscle lateral to and the interspace lateral to both meridians (P<0.05). The ρi and ρe values (Ω•cm) of the ST, GB, the muscle lateral to and the interspace lateral to ST/GB were 658.9±78.5 and 953.8±75.3, 528.0±90.1 and 833.9±101.7, 669.9±71.8 and 1 059.8±86.0, 655.9±64.8 and 1 099.3±93.3, respectively. The ρi and ρe values were significantly lower at the GB Meridian Than at the other three locntions, and the ρe value of ST Meridian was significantly lower than those of the muscle lateral to and the interspace lateral to ST/GB meridians (P<0.01)．. CONCLUSION: The Ri, Re, ρi and ρe values of the ST and GB meridians are significantly lower than those of their neighboring tissues at the same levels of the lower leg, suggesting a more extracellular fluid in the meridian running course and providing evidence for our speculation that the meridian is a hydraulic resistance channel.