Historical trajectories of antibiotics resistance genes assessed through sedimentary DNA analysis of a subtropical eutrophic lake.

Investigating the occurrence of antibiotic-resistance genes (ARGs) in sedimentary archives provides opportunities for reconstructing the distribution and dissemination of historical (i.e., non-anthropogenic origin) ARGs. Although ARGs in freshwater environments have attracted great attention, historical variations in the diversity and abundance of ARGs over centuries to millennia remain largely unknown. In this study, we investigated the vertical change patterns of bacterial communities, ARGs and mobile genetic elements (MGEs) found in sediments of Lake Chenghai spanning the past 600 years. Within resistome preserved in sediments, 177 ARGs subtypes were found with aminoglycosides and multidrug resistance being the most abundant. The ARG abundance in the upper sediment layers (equivalent to the post-antibiotic era since the 1940s) was lower than those during the pre-antibiotic era, whereas the ARG diversity was higher during the post-antibiotic era, possibly because human-induced lake eutrophication over the recent decades facilitated the spread and proliferation of drug-resistant bacteria. Statistical analysis suggested that MGEs abundance and the bacterial community structure were significantly correlated with the abundance and diversity of ARGs, suggesting that the occurrence and distribution of ARGs may be transferred between different bacteria by MGEs. Our results provide new perspectives on the natural history of ARGs in freshwater environments and are essential for understanding the temporal dynamics and dissemination of ARGs.

Effects of Tremella fuciformis Mushroom Polysaccharides on Structure, Pasting, and Thermal Properties of Chinese Chestnuts (Castanea henryi) Starch Granules under Different Freeze-Thaw Cycles.

The purpose of this study was to investigate the effect of Tremella fuciformis polysaccharides on the physicochemical properties of freeze-thawed cone chestnut starch. Various aspects, including water content, crystallinity, particle size, gelatinization, retrogradation, thermal properties, rheological properties, and texture, were examined. The results revealed that moderate freezing and thawing processes increased the retrogradation of starch; particle size, viscosity, shear type, hinning degree, and hardness decreased. After adding Tremella fuciformis polysaccharide, the particle size, relative crystallinity, and gelatinization temperature decreased, which showed solid characteristics. Consequently, the inclusion of Tremella fuciformis polysaccharide effectively countered dehydration caused by freezing and thawing, reduced viscosity, and prevented the retrogradation of frozen-thawed chestnut starch. Moreover, Tremella fuciformis polysaccharide played a significant role in enhancing the stability of the frozen-thawed chestnut starch. These findings highlight the potential benefits of incorporating Tremella fuciformis polysaccharides in starch-based products subjected to freeze-thaw cycles.

[Characteristics of Industrial Volatile Organic Compounds(VOCs) Emission in China from 2011 to 2019].

In order to better understand the industrial volatile organic compounds(VOCs) emissions in China in recent years, an industrial VOCs emission inventory was developed from 2011 to 2019, based on the dynamic emission factors method and the comprehensive source classification system. The results showed that VOCs emissions increased first from 11122.7 kt in 2011 to 13397.9 kt in 2017, and then decreased to 13247.0 kt in 2019. The emission structure of the four source categories changed. The contribution from basic organic chemical industries, gasoline storage and transportation, manufacturing(i.e., coatings, inks, pigments, and similar products), and industrial protective coatings continued to increase. On the contrary, the contributions of oil and natural gas processing, automobile, and container manufacturing industries declined over the study period. Among the industrial emissions in China in 2019, industrial coating, printing, and basic organic chemical industries emitted large amounts of VOCs(accounting for 39.2% of the total emission), and because their contribution became increasingly prominent since 2011, these sectors will be the key emission sources in the future. With respect to the spatial distribution in 2019, East China and South China had the largest VOCs emissions. Shandong, Guangdong, Jiangsu, and Zhejiang were the four provinces that contributed the most, accounting for 40.6% of the total VOCs emissions.

Freshwater lake ecosystem health assessment and its response to pollution stresses based on planktonic index of biotic integrity.

Based on the planktonic data monitored in Lake Balihe, an ecosystem health assessment system referring to planktonic index of biotic integrity (P-IBI) was constructed to evaluate the lake health status and study the response mechanism of P-IBI to environmental factors. The results showed that a total of 7 phylum 59 species of phytoplankton and 3 phylum 28 species of zooplankton were identified in the lake, and both the plankton density and biomass varied significantly in time and space scales. The significant variation of Protozoa density should be responsible to the inconsistency between the distributions of zooplankton density and biomass, as well as the lowest value of P-IBI in summer. The P-IBI values and therefore the health levels can be seasonally ranked as winter > autumn > spring > summer and found spatially increased along the flow direction. Based on the relationships between P-IBI and the environmental factors, ammonia nitrogen = 0.46 mg/L and Secchi depth = 63 cm were found as the environmental protection thresholds of planktonic biotic integrity for this freshwater lake ecosystem. The findings of the research may provide some guidance to the ecological monitoring and protection of freshwater lake.

Analyses on the Temporal and Spatial Characteristics of Water Quality in a Seagoing River Using Multivariate Statistical Techniques: A Case Study in the Duliujian River, China.

In the Duliujian River, 12 water environmental parameters corresponding to 45 sampling sites were analyzed over four seasons. With a statistics test (Spearman correlation coefficient) and multivariate statistical methods, including cluster analysis (CA) and principal components analysis (PCA), the river water quality temporal and spatial patterns were analyzed to evaluate the pollution status and identify the potential pollution sources along the river. CA and PCA results on spatial scale revealed that the upstream was slightly polluted by domestic sewage, while the upper-middle reach was highly polluted due to the sewage from feed mills, furniture and pharmaceutical factories. The middle-lower reach, moderately polluted by sewage from textile, pharmaceutical, petroleum and oil refinery factories as well as fisheries and livestock activities, demonstrated the water purification role of wetland reserves. Seawater intrusion caused serious water pollution in the estuary. Through temporal CA, the four seasons were grouped into three clusters consistent with the hydrological mean, high and low flow periods. The temporal PCA results suggested that nutrient control was the primary task in mean flow period and the monitoring of effluents from feed mills, petrochemical and pharmaceutical factories is more important in the high flow period, while the wastewater from domestic and livestock should be monitored carefully in low flow periods. The results may provide some guidance or inspiration for environmental management.

Analyzing the significant environmental factors on the spatial and temporal distribution of water quality utilizing multivariate statistical techniques: a case study in the Balihe Lake, China.

The assessment of surface water quality is significant to the management of aquatic ecosystem. In this research, in Balihe Lake which is an agricultural watershed lake, 11 environmental parameters (pH, water temperature, water depth, turbidity, DO, COD, TN, NH4+-N, NO3--N, TP, Chl-a) are monitored at 45 sampling sites in four seasons (winter of 2016, spring, summer, and autumn of 2017). With these monitoring data, two kinds of multivariate statistical methods including cluster analysis (CA) and principal component analysis (PCA) are applied to evaluate the spatial and temporal characteristics of the surface water quality. The results reveal that the spatial clusters (less, moderately, and highly polluted sections) of 45 sampling sites classified by the CA method are exactly consistent with the geographical distribution of these sampling sites, which rely on water quality meliorating downstream. From the perspective of time scale, the correlations between environmental parameters generated by the PCA method reveal that the main factors affecting the surface water quality are different in the four seasons. For the whole study period, which is a longer time scale rather than season, the main factors are also different to that of any season. Large time scale may weaken the effect and potential risk of nutrients on water quality, and it is therefore reasonable to select seasonal scale for the study of water quality in an agricultural watershed by using PCA. The results of this research may demonstrate significance to the identification of the main pollution factors and water quality assessment in freshwater lake with multivariate statistical methods.

The Cardiovascular Effect of Systemic Homocysteine Is Associated with Oxidative Stress in the Rostral Ventrolateral Medulla.

It has been demonstrated that homocysteine (HCY) is a significant risk factor of hypertension, which is characterized by overactivity of sympathetic tone. Excessive oxidative stress in the rostral ventrolateral medulla (RVLM), a key region for control of sympathetic outflow, contributes to sympathetic hyperactivity in hypertension. Therefore, the goal of the present study is to determine the effect of systemic HCY on production of reactive oxygen species (ROS) in the RVLM. In the rat model of the diet-induced hyperhomocysteinemia (L-methionine, 1 g/kg/day, 8 weeks), we found that the HCY resulted in a significant increase (≈3.7-fold, P < 0.05) in ROS production in the RVLM, which was paralleled with enhanced sympathetic tone and blood pressure (BP). Compared to the vehicle group, levels of BP and basal renal sympathetic nerve activity in the HCY group were significantly (P < 0.05, n = 5) increased by an average of 27 mmHg and 31%, respectively. Furthermore, the rats treated with L-methionine (1 g/kg/day, 8 weeks) showed an upregulation of NADPHase (NOX4) protein expression and a downregulation of superoxide dismutase protein expression in the RVLM. The current data suggest that central oxidative stress induced by systemic HCY plays an important role in hypertension-associated sympathetic overactivity.

Nandrolone attenuates aortic adaptation to exercise in rats.

AIMS: In this study, we investigated the interaction between exercise-induced mitochondrial adaptation of large vessels and the effects of chronic anabolic androgenic steroids (AASs). METHODS AND RESULTS: Four groups of Sprague-Dawley rats were studied: (i) sedentary, (ii) sedentary + nandrolone-treated, (iii) aerobic exercise trained, and (iv) trained + nandrolone-treated. Aerobic training increased the levels of aortic endothelial nitric oxide synthase (eNOS) and heme oxygenase-1 (HO-1) in accordance with improved acetylcholine-induced vascular relaxation. These beneficial effects were associated with induction of mitochondrial complexes I and V, increased mitochondrial DNA copy number, and greater expression of transcription factors involved in mitochondrial biogenesis/fusion. We also observed enhanced mitochondrial autophagy pathway activity, including increased conversion of LC3-I to LC3-II and greater expression of beclin1 and autophagy-related protein-7 (ATG7). The levels of thiobarbituric acid-reactive substances and protein carbonyls remained unchanged, whereas significant increases in catalase and mitochondrial manganese superoxide dismutase (MnSOD) levels were observed in the aortas of trained animals, when compared with sedentary controls. Nandrolone increased oxidative stress biomarkers and inhibited exercise-induced increases of eNOS, HO-1, catalase, and MnSOD expression. In addition, it also attenuated elevated peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and mitofusin-2 expression, and further up-regulated LC3II conversion, beclin1, ATG7, and dynamin-related protein-1 expression. CONCLUSION: These results demonstrate that nandrolone attenuates aortic adaptations to exercise by regulating mitochondrial dynamic remodelling, including down-regulation of mitochondrial biogenesis and intensive autophagy.

Differential changes of aorta and carotid vasodilation in type 2 diabetic GK and OLETF rats: paradoxical roles of hyperglycemia and insulin.

We investigated large vessel function in lean Goto-Kakizaki diabetic rats (GK) and Otsuka Long-Evans Tokushima Fatty diabetic rats (OLETF) with possible roles of hyperglycemia/hyperosmolarity and insulin. Both young and old GK showed marked hyperglycemia with normal insulin level and well-preserved endothelium-dependent and endothelium-independent vasodilation in aorta and carotid artery. There were significant elevations in endothelial/inducible nitric oxide synthase (eNOS/iNOS) and inducible/constitutive heme oxygenase (HO-1/HO-2) in GK. The endothelium-dependent vasodilation in GK was inhibited partly by NOS blockade and completely by simultaneous blocking of HO and NOS. In contrast, OLETF showed hyperinsulinemia and mild hyperglycemia but significant endothelium dysfunction beginning at early ages with concomitantly reduced eNOS. Insulin injection corrected hyperglycemia in GK but induced endothelium dysfunction and intima hyperplasia. Hyperglycemia/hyperosmolarity in vitro enhanced vessel eNOS/HO. We suggest that hyperinsulinemia plays a role in endothelium dysfunction in obese diabetic OLETF, while hyperglycemia/hyperosmolarity-induced eNOS/HO upregulation participates in the adaptation of endothelium function in lean diabetic GK.

Paradoxical effects of streptozotocin-induced diabetes on endothelial dysfunction in stroke-prone spontaneously hypertensive rats.

Although both diabetes and hypertension are risk factors for cardiovascular disease, the role of hyperglycaemia per se in endothelial dysfunction is controversial. This study was designed to examine whether hyperglycaemia, or streptozotocin-induced diabetes, could aggravate endothelial dysfunction in stroke-prone spontaneously hypertensive rats (SHRSP). Hyperglycaemia was induced by streptozotocin in 2-month-old SHRSP and age-matched normotensive Wistar-Kyoto (WKY) rats. The aorta was isolated 8 weeks after induction of hyperglycaemia to record its function and to examine its morphology with transmission electron microscopy. Endothelial/inducible nitric oxide synthase (eNOS/iNOS) and inducible/constitutive haem oxygenase (HO-1/HO-2) levels were determined with Western blotting. Aortic endothelial function and production of reactive oxygen species and nitric oxide were assayed after incubation in vitro in hyperglycaemic, hyperosmolar solution. Streptozotocin-induced diabetes of 8 weeks duration did not result in endothelial dysfunction in normotensive WKY rats. In contrast, hyperglycaemic WKY rats showed significantly enhanced endothelium-dependent vasodilatation, which was abrogated by simultaneous blocking of NOS and HO. The enhanced vasodilatation was associated with elevation of vascular eNOS and HO-1. Significant endothelial dysfunction and massive macrophage-monocyte infiltration were found in SHRSP aorta (the ratio of the number of macrophages to endothelial cells in the intima, expressed as a percentage, was 20.9 ± 2.8% in SHRSP versus 1.9 ± 0.5% in WKY rats, P < 0.01), which was attenuated significantly in hyperglycaemic SHRSP (11.3 ± 1.6%, P < 0.01 versus SHRSP). Acute hyperglycaemia (10 min) aggravated endothelial dysfunction in SHRSP, with a marked increase in intracellular reactive oxygen species and NO production. Sustained in vitro incubation in hyperglycaemic/hyperosmolar conditions (addition of an extra 50 mmol L(-1) of glucose or mannitol to the usual buffer, to produce a final osmolarity of 350 mosmol L(-1)) for 5 h enhanced endothelium-dependent vasodilatation, with elevated vessel NO production and upregulation of eNOS/HO-1 proteins. Sustained hyperglycaemia does not aggravate endothelial dysfunction and macrophage infiltration in SHRSP. Hyperglycaemia/hyperosmolarity-induced upregulation of eNOS and HO-1 may play a role in this paradoxical adaptation of endothelial function.

Biphasic effects of sodium danshensu on vessel function in isolated rat aorta.

AIM: To investigate the effects of sodium danshensu on vessel function in isolated rat aortic ring. METHODS: Thoracic aortae from normal rats were isolated and equilibrated in organ bath with Krebs-Henseleit buffer and ring tension was recorded. Effects of sodium danshensu on basal tonus of the vessel and its effects on vessel contraction and relaxation with or without endothelium were observed. RESULTS: In thoracic arteries under basal tonus, sodium danshensu (0.3-3 g/L) produced a dose-dependent transient contraction. In phenylephrine-precontracted thoracic arteries with or without endothelium, low concentration (0.1-0.3 g/L) of sodium danshensu produced a weak contraction, while high concentrations (1-3 g/L) produced a pronounced vasodilator after a transient vasocontraction. Pre-incubation with sodium danshensu could inhibit vessel contraction induced by phenylephrine and potassium chloride in a concentration-dependent way. Sodium danshensu inhibited phenylephrine- and CaCl(2)-induced vasoconstriction in Ca(2+)-free medium. Pre-incubation with tetraethylammonium, a non-selective K(+) channel blocker, and apamin, a small-conductance calcium-activated K(+) channel blocker partially antagonized the relaxation response induced by sodium danshensu. However, iberiotoxin (big-conductance calcium-sensitive K(+) channel blocker), barium chloride (inward rectifier K(+) channel blocker), and glibencalmide (ATP-sensitive K(+) channel blocker) had no influence on the vasodilation effect of sodium danshensu. CONCLUSION: Sodium danshensu showed a biphasic effects on vessel tension. While low dosage of sodium danshensu produced small contraction possibly through transient enhancement of Ca(2+) influx, high dosage produced significant vasodilation mainly through promoting the opening of non-selective K(+) channels and small-conductance calcium-sensitive K(+) channels in the vascular smooth muscle cells.

Effects of different levels of exercise volume on endothelium-dependent vasodilation: roles of nitric oxide synthase and heme oxygenase.

The objective of this study was to examine the effects of moderate and high levels of exercise volume on endothelium-dependent vasodilation and associated changes in vascular endothelial/inducible nitric oxide synthase (eNOS and iNOS) and heme oxygenase (HO). Male Sprague-Dawley rats were assigned to sedentary control, acute (2 weeks), or chronic (6 weeks) treadmill running at moderate intensity (50% maximal aerobic velocity) with different durations of exercise episodes: 2 h/d (endurance training, moderate volume) and 3 h/d (intense training, high volume). Endothelium-dependent vascular function was examined in isolated thoracic aorta. Co-localization and contents of aortic eNOS/iNOS and HO-1/HO-2 were determined with immunofluorescence and Western blotting. Compared with sedentary controls, rats subjected to acute and chronic endurance training showed enhanced endothelium-dependent relaxation (p<0.01). Whereas acetylcholine-induced dilation was inhibited completely by NOS inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) in sedentary controls, the dilation in the training groups was only partly blocked by L-NAME (inhibition was 98+/-3%, 79+/-6%, and 77+/-5% in sedentary control, acute, and chronic training groups, respectively, p<0.01). The remnant dilation in the training groups was further inhibited by HO inhibitor protoporphyrin IX zinc, with concomitant elevation in aortic eNOS as well as HO-1 and HO-2. In contrast to endurance exercise, high-volume intense training resulted in mild hypertension with significant impairment in endothelium-dependent vasodilation and profuse increases in aortic iNOS and eNOS (p<0.01). In conclusion, endothelium-dependent vasodilation is improved by endurance exercise but impaired by chronic intense training. Elevations of vascular eNOS and HO-1/HO-2 may contribute to enhanced vasodilation, which can be offset by intense training and elevation in vascular iNOS.

Elevated catalase and heme oxygenase-1 may contribute to improved postischaemic cardiac function in long-term type 1 diabetes.

1. Although increased oxidative stress has been shown repeatedly to be implicated in diabetes, the cardiovascular anti-oxidant state and heart response to ischaemia in long-term Type 1 diabetes remain largely unknown. The present study was designed to observe heart tolerance to ischaemia-reperfusion and endogenous anti-oxidants in the cardiovascular system in long-term hyperglycaemic rats. 2. Hearts from Sprague-Dawley rats surviving up to 6 months with streptozocin-induced severe hyperglycaemia (blood glucose > 20 mmol/L) were isolated and subjected to global ischaemia and reperfusion. Cardiac function, electrocardiogram and anti-oxidants in the myocardium and aorta were examined. In addition, the morphology of the myocardial mitochondria and the in vitro function of aortic vessels were assessed. 3. Hearts from diabetic rats demonstrated lower baseline heart function but had higher postischaemic coronary flow and left ventricular developed pressure compared with their respective controls (P < 0.05). In addition, hearts from diabetic animals had fewer arrhythmias (P < 0.01) and lower left ventricular end-diastolic pressure during reperfusion (P < 0.05). Higher catalase and heme oxygenase-1 content was found in the aorta and myocardium from diabetic rats (P < 0.01). In aortas from diabetic animals, acetylcholine-induced vasodilatation was enhanced and was approximately 15% after inhibition of nitric oxide synthase, compared with 0% in controls. The 15% relaxation was abrogated by heme oxygenase blockade. Mitochondria from the myocardium of diabetic rats showed significant increases in both size and number (P < 0.05). 4. Hearts of long-term Type 1 diabetic rats demonstrated improved recovery of postischaemic cardiac function and reduced reperfusion arrhythmia. Hyperglycaemia may enhance cardiovascular anti-oxidant capacity and mitochondrial neogenesis, which renders the heart resistant to ischaemia and oxidative injury.

Mechanisms underlying vasorelaxant action of astragaloside IV in isolated rat aortic rings.

1. Astragaloside IV is a component from the widely used traditional Chinese herb Astragalus membranaceus and its effect on rat aortic ring contraction and relaxation were investigated. 2. The aorta from male Sprague-Dawley rats was isolated in an organ bath and ring tension was recorded with or without endothelium. Cumulative effects of astragaloside IV on vessel contraction and relaxation were observed in the presence of various antagonists related to vessel activity. 3. Astragaloside IV showed concentration-dependent inhibition of vessel contraction induced by phenylephrine and potassium chloride. The amount of calcium released from intracellular stores sensitive to phenylephrine was also markedly reduced by astragaloside IV. There was dose-dependent vasorelaxation in endothelium-intact rings, which was partly inhibited by pre-incubation with nitric oxide (NO) synthase (NOS) Nomega-nitro-L-arginine methyl ester and guanylate cyclase inhibitor, 1H-[1,2,4] oxadiazolo [4,3-alpha] quinoxalin-1-one. Astragaloside IV also induced a significant increase in aortic tissue content of guanosine 3",5"-cyclic monophosphate (cGMP) both in vivo and in vitro. Endothelial NOS inhibitor Nomega-nitro-L-arginine prevented vasodilatation, whereas neuronal NOS inhibitor 7-nitroindazole did not show significant influence on the vessel relaxation of astragaloside IV. 4. In conclusion, astragaloside IV inhibited vessel contraction through blocking calcium influx and intracellular calcium release. The endothelium-dependent vessel dilation of astragaloside IV was attributed mainly to the endothelium-dependent NO-cGMP pathway.

[Correlation between brain-derived neurotrophic factor expression in the nerve system and erectile dysfunction in diabetic rats].

OBJECTIVE: To observe the correlation between brain-derived neurotrophic factor (BDNF) expression in the nerve system of diabetes mellitus (DM) rats and diabetic erectile dysfunction (ED). METHODS: DM rats were induced by injecting streptozotocin and erectile function test was done by injecting apomorphine (APO) at 1 month, 2 months, 3 months and 4 months. Then the brain, lumbosacral spinal cord, thoracic and lumbar sympathetic trunks, penis and prostate were taken from the diabetic and normal rats of the same age. The BDNF positive neurons and nerve fibers were shown by immunohistochemistry or fluorescence immunohistochemistry. The number and the grey density of BDNF positive cells and fibers were detected by image analysis. RESULTS: Compared with the control group, the erection frequency of the DM rats decreased at 2 months (P <0. 05) , and significantly at 3 and 4 months (P > 0.01) , and the BDNF positive neurons and nerve fibers in the cerebral cortex, lumbosacral spinal cord, thoracic and lumbar sympathetic trunks, penis and prostate of 1-month DM rats were reduced (P <0. 05). As time went on, BDNF declined progressively. CONCLUSION: BDNF decreases in the central and peripheral nerve system in the early stage of diabetes mellitus, wich is closely correlated with diabetic ED.

Expression of fibroblast growth factors in rat dorsal root ganglion neurons and regulation after peripheral nerve injury.

Using cDNA array, we observed the expression of eight members of the fibroblast growth factor (FGF) family, FGF 2, 5, 7, 9, 10, 13 and 14, in rat lumbar 4 and 5 dorsal root ganglia (DRGs). Over a period of 28 days after sciatic nerve transection, the array signals for FGF 2 and 7 were significantly increased in the DRGs, while FGF 13 decreased. Using the reverse transcription polymerase chain reaction (RT-PCR), we confirmed the axotomy-induced changes in the expression of FGF 7 and 13. hybridization showed that FGF 13 was expressed in 60% of DRG neurons under normal circumstance. Seven days after axotomy the number of FGF 13-positive neurons was decreased to 18%, but partially recovered to 40% after 28 days. FGF 13 immunoreactivity was also decreased. These data indicate that FGFs are important for DRG neurons under normal circumstance and after nerve injury.