Mechanism of static loading injury in human skeletal muscle cells.

OBJECTIVE: To establish a cellular-level mechanical injury model for human skeletal muscle cells and investigate changes in the mechanical effect mechanism after such injuries. METHODS: The FX-5000™ Compression System was used to apply constant static mechanical pressure to human skeletal muscle cells. A factorial design analysis was conducted to discover the optimal injury model by evaluating the correlation between the amount of pressure, the duration of mechanical stimulation, and the number of days of observation. Skeletal muscle cell injury was evaluated by measuring cell metabolism, morphology, and calcium homeostasis. RESULTS: Mechanical injury was modeled as continuous pressure of 1 MPa for 2 hours with observation for 3 days. The results show that mechanical injury increased creatine kinase, intracellular Ca2+ concentration, and malondialdehyde content, decreased superoxide dismutase, and caused cell swelling and severe cytoplasmic vacuolization (all P < 0.05). CONCLUSION: This model of mechanically-injured human skeletal muscle cells provides an experimental model for the clinically common skeletal muscle injury caused by static loading pressure. It may be used to study the mechanism of action of treatment methods for mechanically injured skeletal muscle.

OsCOPT7 is a copper exporter at the tonoplast and endoplasmic reticulum and controls Cu translocation to the shoots and grain of rice.

Copper (Cu) is an essential micronutrient for all living organisms but is also highly toxic in excess. Cellular homoeostasis of Cu is maintained by various transporters and metallochaperones. Here, we investigated the biological function of OsCOPT7, a member of the copper transporters (COPT) family, in Cu homoeostasis in rice. OsCOPT7 was mainly expressed in the roots and the expression was upregulated by Cu deficiency. OsCOPT7 was localized at the tonoplast and the endoplasmic reticulum. Knockout of OsCOPT7 increased Cu accumulation in the roots but decreased Cu concentrations in the shoots and grain. The knockout mutants contained higher concentrations of Cu in the roots cell sap but markedly lower concentrations of Cu in the xylem sap than wild-type plants. Seed setting and grain yield were reduced significantly in the knockout mutants grown in a low Cu soil. Knockout mutants were more tolerant to Cu toxicity. Yeast two-hybrid and bimolecular fluorescence complementation assays showed that OsCOPT7 interacts physically with the rice Cu chaperone antioxidant protein 1 (OsATX1). Taken together, our results indicate that OsCOPT7 is a specific Cu transporter functioning to export Cu from the vacuoles and the ER and plays an important role in controlling the root-to-shoot Cu translocation in rice.

Study of polycyclic aromatic hydrocarbons accumulation in bioretention facilities and its influence on microbial community structure.

Bioretention facilities are one of the most widely used measures for urban stormwater control and utilization. In this study, the accumulation characteristics of polycyclic aromatic hydrocarbons (PAHs) in bioretention facilities and the effects of PAHs on the structure of microbial communities were explored by combining on-site monitoring and water distribution simulation experiments. The correlation between pollutant accumulation and dominant microorganisms in the bioretention systems was also clarified. The results showed that all 16 priority PAHs were detected in the bioretention facilities in the sponge city pilot area. The PAH concentrations in the soil during the non-rainy season were higher than those in the rainy season and medium- and high-ring PAHs dominated. PAHs in the study area were mainly derived from coal and biomass combustion. The potential carcinogenic risk of PAHs accumulated in the bioretention facilities in the study area was low. The microbial diversity during the non-rainy season was greater than that during the rainy season. Firmicutes, Bacteroidetes, Bacteroides, and Massilia were strongly correlated with naphthalene (NAP), pyrene (PYR), fluoranthene (FLT), and benzo[a]pyrene (BaP). According to the results of the small-scale water distribution test, the addition of PAHs had little effect on the decline in water quantity, and there was no significant regularity in the reduction of water quality including TP, NH4+-N, NO3-N, and TN. The addition of PAHs had a significant effect on the microbial community structure and an inhibitory effect on enzyme activity.

Study on non-point source pollution characteristics under different spatial scales: a case study of Hanjiang River Basin, China.

In recent years, the research on non-point source (NPS) pollution has been deepening, but it is focused on the large-scale watershed or region. There are a few studies on the scales of small watershed and runoff plot, and it is even less to analyze the characteristics and mechanism of non-point source pollution in certain watershed by combining three different scales. Based on the combination of natural rainfall monitoring and MIKE model simulation, the Shaanxi section of Hanjiang River Basin in China was taken as an example to study the characteristics of NPS pollution at different spatial scales. The results showed that there was an obvious correlation between rainfall and runoff/sediment yield. The order of runoff yield/sediment yield per unit area was as follows: woodland > forested and grassy land > arable land. There was a significant relationship between the loss of total phosphorus and the sediment yield in the runoff plots. The total nitrogen pollution was serious, with an average concentration of 3.8 mg/L. The nutrient loss was in the form of nitrate nitrogen, with an average proportion of 63.06%. For small watershed scale, the characteristics of rainfall runoff pollution generation were like runoff plot scale, both have obvious initial scour phenomenon. However, compared with runoff plot scale, the pollutant loss concentration increases with a strong lag. The MIKE model based on the coupling of hydrology, hydrodynamics, and pollution load had a strong applicability in the basin. The critical source areas of NPS pollution were identified, and five scenarios were laid out in the areas for the control of NPS pollution. Centralized livestock and poultry farming had the best reduction effect.

Rapid identification of high and low cadmium (Cd) accumulating rice cultivars using machine learning models with molecular markers and soil Cd levels as input data.

Excessive accumulation of cadmium (Cd) in rice grains threatens food safety and human health. Growing low Cd accumulating rice cultivars is an effective approach to produce low-Cd rice. However, field screening of low-Cd rice cultivars is laborious, time-consuming, and subjected to the influence of environment × genotype interactions. In the present study, we investigated whether machine learning-based methods incorporating genotype and soil Cd concentration can identify high and low-Cd accumulating rice cultivars. One hundred and sixty-seven locally adapted high-yielding rice cultivars were grown in three fields with different soil Cd levels and genotyped using four molecular markers related to grain Cd accumulation. We identified sixteen cultivars as stable low-Cd accumulators with grain Cd concentrations below the 0.2 mg kg-1 food safety limit in all three paddy fields. In addition, we developed eight machine learning-based models to predict low- and high-Cd accumulating rice cultivars with genotypes and soil Cd levels as input data. The optimized model classifies low- or high-Cd cultivars (i.e., the grain Cd concentration below or above 0.2 mg kg-1) with an overall accuracy of 76%. These results indicate that machine learning-based classification models constructed with molecular markers and soil Cd levels can quickly and accurately identify the high- and low-Cd accumulating rice cultivars.

[The Effect and Mechanism of Novel Telomerase Inhibitor Nilo 22 on Leukemia Cells].

OBJECTIVE: To investigate the cytotoxic effect and its mechanism of the micromolecule compound on the leukemia cells. METHODS: The cytotoxic effects of 28 Nilotinib derivatives on K562, KA, KG, HA and 32D cell lines were detected by MTT assays, and the compound Nilo 22 was screen out. Cell apoptosis and cell cycle on leukemia cells were detected by flow cytometry. The effect of compound screened out on leukemogenesis potential of MLL-AF9 leukemia mice GFP+ cells was tested by colony-forming units assays (CFU). The cytotoxic effect was further detected by transplant assays ex vivo. Telomerase activity assay, C-circle assay were used to measure the effects of compound on the length mechanism of telomere, RT-PCR was used to detected the changes of telomere. RESULTS: Nilo 22 serves as the most outstanding candidate out of 28 Nilotinib derivatives, which impairs leukemia cell lines, but spares normal hematopoietic cell line. Comparing with Nilotinib, Nilo 22 could induce the apoptosis of GFP+ cells significantly, slightly arrests the cell cycle at G0/G1 phase, and significantly inhibits colony formation and prolong the progression in MLL-AF9 leukemia mice model. The expression showed that the compound could slow the disease progression in MLL-AF9 leukemia mice significantly. Mechanistically, Nilo 22 could reduce the length of telomere by inhibiting telomerase activity and alternative lengthening of telomere (ALT). CONCLUSION: Nilo 22 shows a significant cytotoxic effect on mice and human leukemia cells, especially for drug resistance cells. Nilo 22 is a promising anti-leukemia agent to solve the common clinical problems of drug resistance and relapse of leukemia.

Intermittent pressure imitating rolling manipulation ameliorates injury in skeletal muscle cells through oxidative stress and lipid metabolism signalling pathways.

BACKGROUND: Traditional Chinese medicine manipulation (TCMM) is often used to treat human skeletal muscle injury, but its mechanism remains unclear due to difficulty standardizing and quantifying manipulation parameters. METHODS: Here, dexamethasone sodium phosphate (DSP) was utilized to induce human skeletal muscle cell (HSkMC) impairments. Cells in a three-dimensional environment were divided into the control normal group (CNG), control injured group (CIG) and rolling manipulation group (RMG). The RMG was exposed to intermittent pressure imitating rolling manipulation (IPIRM) of TCMM via the FX­5000™ compression system. Skeletal muscle damage was assessed via the cell proliferation rate, superoxide dismutase (SOD) activity, malondialdehyde (MDA) content and creatine kinase (CK) activity. Isobaric tagging for relative and absolute protein quantification (iTRAQ) and bioinformatic analysis were used to evaluate differentially expressed proteins (DEPs). RESULTS: Higher-pressure IPIRM ameliorated the skeletal muscle cell injury induced by 1.2 mM DSP. Thirteen common DEPs after IPIRM were selected. Key biological processes, molecular functions, cellular components, and pathways were identified as mechanisms underlying the protective effect of TCMM against skeletal muscle damage. Some processes (response to oxidative stress, response to wounding, response to stress and lipid metabolism signalling pathways) were related to skeletal muscle cell injury. Western blotting for 4 DEPs confirmed the reliability of iTRAQ. CONCLUSIONS: Higher-pressure IPIRM downregulated the CD36, Hsp27 and FABP4 proteins in oxidative stress and lipid metabolism pathways, alleviating excessive oxidative stress and lipid metabolism disorder in injured HSkMCs. The techniques used in this study might provide novel insights into the mechanism of TCMM.

Nonmetric multidimensional scaling and adverse effects on aquatic biota of polycyclic aromatic hydrocarbons in sediments: A case study of a typical aquaculture wetland, China.

Aquaculture wetlands, particularly those located within urban areas, are fragile ecosystems which are generally heavily impacted by urban and aquaculture activities. However, there are, to date, no reports on adverse effects related to polycyclic aromatic hydrocarbons (PAHs) in sediments from aquaculture wetlands in metropolitan areas. The characterization, sources, and incidence of adverse effects on aquatic biota of PAHs were studied in the riverine and estuarine sediments of the Rongjiang River in an aquaculture wetland in Chaoshan metropolis. The total PAH concentrations varied from 46.0 to 1665.2 ng/g (dry weight), with a mean of 246.4 ng/g, indicating lower concentrations than those of other comparable rivers and estuaries worldwide. High-molecular-weight PAHs were the major PAH species, with fluorene, benzo(b)fluoranthene, and benzo(a)pyrene as the dominant components. These PAHs are likely to be mainly derived from oil and coal/biomass combustion. Probability risk assessment indicated a moderate PAH ecological risk in the Rongjiang River and its estuary (RJE). Accordingly, a comprehensive management plan should be established to ensure sustainable aquaculture wetlands.

Differential Strategies to Tolerate Flooding in Polygonum hydropiper Plants Originating From Low- and High-Elevation Habitats.

In species that occur over a wide range of flooding conditions, plant populations may have evolved divergent strategies as a consequence of long-term adaptation to local flooding conditions. In the present study, we investigated the effects of a flooding gradient on the growth and carbohydrate reserves of Polygonum hydropiper plants originating from low- and high-elevation habitats in the Dongting Lake wetlands. The results indicated that shoot length did not differ, whereas the total biomass and carbohydrate reserves were reduced under flooded compared to well-drained conditions for plants originating from both habitat types. However, shoot length, shoot mass, rhizome mass, and total biomass were lower in plants from low-elevation habitats than in those from high-elevation habitats in the flooded condition. Soluble sugar and starch contents in belowground biomass were higher in plants from low-elevation habitats than in those from high-elevation habitats independently of the water level. Therefore, P. hydropiper plants from low-elevation habitats exhibit a lower growth rate and more conservative energy strategy to cope with flooding in comparison with plants from high-elevation habitats. Differential strategies to cope with flooding among P. hydropiper populations are most likely a response to the flooding pressures of the habitat of origin and may potentially drive ecotype differentiation within species along flooding gradients.

Changes of the Macrobenthic Faunal Community with Stand Age of a Non-native Mangrove Species in Futian Mangrove National Nature Reserve, Guangdong, China.

Ya-Fang Li, Fei-Yan Du, Yang-Guang Gu, Jia-Jia Ning, and Liang-Gen Wang (2017) Sonneratia apetala, a non-native superior rapidly growing mangrove species with wide environmental tolerance, has been introduced to Futian National Nature Reserve in Shenzhen, Guangdong, China, for mangrove restoration since 1993. However, the community structure of the associated macrobenthic fauna, a vital component of energy ow and nutrient recycling, remains obscure. The present study analyzed the macrobenthic faunal community, associated habitat characteristics and physico-chemical properties of sediment in rehabilitated S. apetala forests at stand ages of 8, 9, 14, 16 and 20 years from November 2014 to May 2015. Habitat complexity and stand structural heterogeneity varied with stand age. Sediment physico-chemical properties were similar for all stands analyzed, although soil organic matter (SOM) content was significantly higher in the 20-year-old stand than in others. Shannon-Weaver (H') and Pielou's evenness (J) indices of macrobenthic fauna were highest in 14- and 16-year- old stands, respectively, and lowest in 8-year-old stands. In contrast, abundance and biomass peaked in 8-year- old stands and were lowest in 16-year-old stands. Multivariate analysis (cluster, ANOSIM and SIMPER) showed that the macrobenthic faunal community in the 20-year-old stand was different from other stand ages because of a greater abundance of small-sized mollusks and opportunistic species. Spearman correlation analysis showed that H' was positively correlated with salinity. The distance-based linear model suggested that SOM was a significant predictor variable correlated with the macrobenthic faunal community. However, SOM was the only significant predictor variable explaining 12.7% of the total variation; this implies that the spatial variation of the macrobenthic faunal community here was mostly independent of the sediment properties measured. Therefore, we conclude that habitat characteristics such as vegetation characteristics can potentially explain the majority of the variation.

Fuzzy comprehensive assessment of heavy metals and Pb isotopic signature in surface sediments from a bay under serious anthropogenic influences: Daya Bay, China.

The fuzzy comprehensive assessment was undertaken to evaluate the marine sediment quality in Daya Bay, China based on the monitoring data of 2015 and China National Standard for Marine Sediment Quality (GB 18668-2002). The results demonstrated that the average metal concentrations (mg/kg) were 0.08 (Cd), 51.30 (Pb), 91.30 (Cr), 29.63 (Cu), 143.42 (Zn), 0.03 (Hg), and 7.31 (As), which were clearly higher with respect to their corresponding background values. Cr was the major pollutant based weight matrices calculated. Taken as whole, the membership degree of class I was a range from 0.65 to 1, suggesting that the marine sediment quality in Daya Bay belongs to class I. Pb stable isotopic analysis coupled with geographic information system (GIS) approach showed the sedimentary Pb mainly originated from petrochemical industry at northwest coast of Daya Bay.

Trade-off between allocation to reproductive ramets and rhizome buds in Carex brevicuspis populations along a small-scale elevational gradient.

The trade-off between allocation to sexual and clonal reproduction in clonal plants is influenced by a variety of environmental factors; however, it has rarely been examined under field conditions. In this study, we investigated the trade-off between two modes of reproduction in Carex brevicuspis C. B. Clarke across a small-scale elevational gradient (21-27 m a.s.l.) at the Dongting Lake wetlands, China. The proportion of biomass allocated to and the density of reproductive ramets were higher at low than at intermediate and high elevations. In contrast, the proportion of biomass allocated to and the density of rhizome buds were lower at low than at intermediate and high elevations. Redundancy analysis showed that sexual reproduction was positively correlated with soil moisture content, soil organic matter, total phosphorus, and pH, and negatively correlated with elevation and ramet density. Our findings suggested that allocation to sexual reproduction is favored in disturbed habitats with fertile soils, whereas allocation to vegetative propagation is favored in stable and competitive habitats. Trade-off between allocation to sexual reproduction and vegetative propagation along an elevational gradient might be a reproductive strategy of C. brevicuspis to adapt to the water level fluctuations in wetland habitats.

Optimization of ultrasound extraction of Alisma orientalis polysaccharides by response surface methodology and their antioxidant activities.

Rhizoma alismatis (the rhizome of Alisma orientalis) polysaccharides (RAP) have been reported to have a variety of important biological activities. However, effective extraction of RAP has been an unsolved issue. In this study, we used an ultrasound method for high yield extraction of RAP and optimized the conditions using the response surface methodology (RSM). Following multiple regression analyses of the experimental results, we applied the 3-D response surface and the contour plots to determine the optimal conditions, which were found to be ultrasound treatment at 76.1°C for 75.2 min, and water to material ratio at 30.1 ml/g. Under such conditions, the yield was 6.90% which was much higher than traditional hot water extraction yield (3.41%). The fractionated RAPs following stepwise ethanol precipitation showed strong antioxidant activities. The results indicated that ultrasound extraction was a very effective method for the extraction of RAP and the polysaccharides could be explored as a potential antioxidant agent for use in medicine or functional food.

The role of peritoneal alternatively activated macrophages in the process of peritoneal fibrosis related to peritoneal dialysis.

It has been confirmed that alternatively activated macrophages (M2) participate in tissue remodeling and fibrosis occurrence, but the effect of M2 on peritoneal fibrosis related to peritoneal dialysis (PD) hasn't been elucidated. This study was therefore conducted to assess the association between M2 and peritoneal fibrosis related to PD. In this study, peritoneal fibrosis was induced by intraperitoneal (i.p.) injection of Lactate-4.25% dialysate (100 mL/kg) to C57BL/6J mice for 28 days, and liposome-encapsulated clodronate (LC, the specific scavenger of macrophages) was used to treat the peritoneal fibrosis mice model by i.p. injection at day 18 and day 21. All animals were sacrificed at day 29. Parietal peritonea were stained with Masson's trichrome, and the expression of type I collagen (Col-I), fibronectin, mannose receptor (CD206), transforming growth factor beta (TGF-ß), chemokine receptor 7 (CCR7), chitinase 3-like 3 (Ym-1) and arginase-1 (Arg-1) was determined by Western blotting, immunofluorescence and quantitative real-time PCR. Our results revealed that peritoneal thickness, Col-I, fibronectin, CD206, TGF-ß, Ym-1 and Arg-1 were upregulated in the peritoneal fibrosis mice model, and all of these indexes were downregulated in those treated with LC. Additionally, there was no difference in the level of CCR7 between the model and treatment group. Our study indicated that peritoneal M2 played an important role in the process of peritoneal fibrosis related to PD and might be a potential target for intervention therapy of peritoneal fibrosis.

[Preliminary study on the value of ratio of serum luteinizing hormone/follicle-stimulating hormone in diagnosis of polycystic ovarian syndrome among women with polycystic ovary].

OBJECTIVE: To investigate the value of ratio of luteinizing hormone (LH) to follicle-stimulating hormone (FSH) in diagnosis of polycystic ovarian syndrome (PCOS) among women with polycystic ovary (PCO) and to compare the difference of the diagnostic criteria between the Rotterdam Consensus and the Committee for Reproductive and Endocrine in Japan Society of Obstetrics and Gynecology. METHODS: By means of transvaginal Doppler ultrasound, 195 women with PCO were diagnosed in Nanfang Hospital of Reproductive Medicine Center and compare difference of multiple clinical indexes according to Rotterdam consensus and Japan consensus respectively. In the mean time, the ratio of LH/FSH, the level of LH, testosterone (T) and recevier operating characteristic (ROC) curve were explored to on the value of diagnosis of PCOS. RESULTS: By Rotterdam consensus, 144 women were diagnosed with PCOS and 51 women were non-PCOS, while 111 were identified as PCOS and 84 were non-PCOS according to Japan consensus. LH/FSH in PCOS and non-PCOS were 1.59 ± 0.84 and 0.85 ± 0.47 respectively when based on Rotterdam consensus, and this ratio were 1.87 ± 0.76 in PCOS and 0.78 ± 0.39 in non-PCOS based on Japan consensus. When using LH/FSH to diagnosis PCOS by Rotterdam consensus and Japan consensus, areas under ROC curve are 0.786 and 0.942, respectively. CONCLUSIONS: The ratio of LH/FSH ≥ 1 provide the significant value in the diagnosis of PCOS. The criteria of the Committee for Reproductive and Endocrine in Japan Society of Obstetrics and Gynecology is more suitable for Chinese patients.

Disruption of Cdk5-associated phosphorylation of residue threonine-161 of the delta-opioid receptor: impaired receptor function and attenuated morphine antinociceptive tolerance.

Morphine is the most commonly used and most effective analgesic in the clinic. However, its use is limited by the tolerance. Evidence indicates that the delta-opioid receptor (DOR) is essential for morphine antinociceptive tolerance; however, their underlying mechanisms are poorly understood. Here, we show that cyclin-dependent kinase 5 (Cdk5), activated in morphine antinociceptive tolerance, directly phosphorylates DOR at Thr-161 in DRG neurons. Cdk5 was found to phosphorylate Thr-161 in the second loop of DOR, but not the corresponding residue in the mu-opioid receptor (MOR). Phosphorylation at Thr-161 is required for normal cell surface expression of DOR, and the formation of DOR-MOR heterodimers. Our studies indicated that inhibition of Cdk5 activity or overexpression of a DOR mutant lacking the Cdk5 phosphorylation site displayed relatively low cell surface expression and relatively low abilities to form heterodimers of DOR and MOR; intrathecal delivery of a construct expressing the T161A mutant of DOR attenuated morphine antinociceptive tolerance in rats, suggesting that Thr-161 phosphorylation of DOR contributed to Cdk5-mediated morphine antinociceptive tolerance. Furthermore, an engineered Tat fusion-interfering peptide corresponding to the second intracellular loop of DOR (Tat-DOR-2L), reduced the cell surface expression of DOR, disrupted the formation of DOR-MOR heterodimers, and significantly attenuated the development of morphine antinociceptive tolerance after intrathecal injection. The present study indicates that Cdk5-mediated phosphorylation of DOR at Thr-161 plays a crucial role in the development of morphine tolerance and suggests the possibility of targeting DOR phosphorylation at Thr-161 to attenuate morphine antinociceptive tolerance during pain management.

Serial public cancer screening in China.

Screening of hollow-organ cancers has been carried out for 16 years using the occult blood test. We are currently assessing the result of serial public cancer screening in China. Human cancers can be divided into 3 types: 1) surface cancer (15%), 2) hollow-organ cancer (65%), and 3) deep-seated solid-organ cancer (20%). Through an education program, the general population was taught to self-examine the body surface and to use a self-examination kit to detect any occult blood discharged from the hollow organs. On discovering positive or doubtful results, these subjects were referred to the hospital for a complete clinical checkup. A total of 10,000 people ages 40 to 70 years chosen from the university areas in Beijing were divided in a test group (5,000) and a control group (5,000). This study, which was carried out for 4 years, showed that 79 cancers were discovered in the test group, with an annual cancer detection rate of 482.5/10,000 population and a cancer mortality rate of 36.6/10,000. The control group showed an annual cancer mortality rate of 206/10,000 population. The difference has very marked statistical significance ( p< 0.001). During the same interval, an extensive cancer screening was carried out in the high-incidence areas. The Cancer Institute in Sihui city of Guangdong province discovered, by screening 758 persons in the outpatient department during a 2-year period, 16 cancers with 15 as nasopharyngeal cancer. From 1999 to 2000, another extensive public cancer screening in the high-incidence areas in Jiangsu province discovered 480 cancers from an 80,000 population. Together with the screening test carried out by Qin's self-examination kit before the National Supported Cancer Research Project on a population of 431,000, 1,272 cancers and more than 10,000 precancerous lesions have been detected. The self-examination kit and public cancer screening are highly important in the practice of public cancer screening.