U-shaped association between serum calcium and in-hospital mortality in diabetes patients with congestive heart failure: a cohort study.

Previous studies have reported that the significant association between serum calcium and mortality substantially in patients, especially among those with intensive care unit (ICU). And In diabetes mellitus, congestive heart failure (CHF) is a significant comorbidity. We aim to evaluate the association between serum calcium levels and in-hospital mortality among patients with diabetes and congestive heart failure. The participants in this study were extracted from the Medical Information Mart for Intensive Care IV (MIMIC-IV) database. To scrutinize potential associations between serum calcium levels and in-hospital mortality, a comprehensive analysis encompassing multivariate logistic regression, cubic spline function model, threshold effect analysis, and subgroup analysis was performed. This retrospective cohort study encompassed 7063 patients, among whom the in-hospital mortality stood at 12.2%. In the multivariate logistic regression, adjusted odds ratios (ORs) were contrasted with the reference category Q6 (8.8-9.1 mg/dL) for serum calcium levels and in-hospital mortality. The adjusted ORs for Q1 (≤ 7.7 mg/dL), Q2 (7.7-8 mg/dL), and Q7 (≥ 9.1 mg/dL) were 1.69 (95% CI 1.17-2.44, p = 0.005), 1.62 (95% CI 1.11-2.36, p = 0.013), and 1.57 (95% CI 1.1-2.24, p = 0.012) respectively. The dose-response analysis uncovered a U-shaped relationship between serum calcium levels and in-hospital mortality in diabetic patients with heart failure. Subgroup analyses confirmed result stability notwithstanding the influence of diverse factors. Our investigation revealed a U-shaped correlation between serum calcium levels and in-hospital mortality in diabetes patients with congestive heart failure, pinpointing a significant inflection point at 9.05 mg/dL.

Single-Cell Transcriptome Analysis of Peripheral Neutrophils From Patients With Idiopathic Pulmonary Arterial Hypertension.

BACKGROUND: Idiopathic pulmonary hypertension (IPAH) is a rare and devastating disease often accompanied by persistent inflammation and immune responses. We aim to provide a reference atlas of neutrophils to facilitate a better understanding of cellular phenotypes and discovery of candidate genes. METHODS: Peripheral neutrophils from naive patients with IPAH and matched controls were profiled. Whole-exon sequencing was performed to exclude known genetic mutations before establishing single-cell RNA sequencing. Marker genes were validated by flow cytometry and histology in a separate validation cohort. RESULTS: Seurat clustering analysis revealed that the landscape of neutrophils encompassed 5 clusters, including 1 progenitor, 1 transition, and 3 functional clusters. The intercorrelated genes in patients with IPAH were mainly enriched in antigen processing presentation and natural killer cell mediated cytotoxicity. We identified and validated differentially upregulated genes, including MMP9 (matrix metallopeptidase 9), ISG15 (ISG15 ubiquitin-like modifier), and CXCL8 (C-X-C motif ligand 8). The positive proportions and fluorescence quantification of these genes were significantly increased in CD16+ neutrophils in patients with IPAH. The higher proportion of positive MMP9 neutrophils increased mortality risk after adjustment for age and sex. Patients with higher proportions of positive MMP9 neutrophils had worse survival, while the fraction of ISG15- or CXCL8-positive expression neutrophils failed to predict outcome. CONCLUSIONS: Our study yields a comprehensive dataset of the landscape of neutrophils in patients with IPAH. The predictive values of a neutrophil cluster characterized by higher MMP9 expression indicate a functional role for neutrophil-specific matrix metalloproteinases in the pathogenesis of pulmonary arterial hypertension.

[Simulation of Anthropogenic CO2 Emissions in the Yangtze River Delta Based on Different Emission Inventories].

Nowadays, great uncertainty still exists on the urban- and regional-scale anthropogenic CO2 emission estimation based on emission inventories. In order to achieve the carbon peaking and neutrality targets for China, it is urgent to accurately estimate anthropogenic CO2 emissions at regional scales, especially in large urban agglomerations. Using two inventories (EDGAR v6.0 inventory and a modified inventory combining EDGAR v6.0 with GCG v1.0) as prior anthropogenic CO2 emission datasets andtaking themas input data respectively, this study utilized the WRF-STILT atmospheric transport model to simulate atmospheric CO2 concentration in the Yangtze River Delta region from December 2017 to February 2018. The simulated atmospheric CO2 concentrations were further improved by referencing atmospheric CO2 concentration observation at a tall tower in Quanjiao County of Anhui Province and using the scaling factors obtained from the Bayesian inversion method. An estimation of anthropogenic CO2 emission flux in the Yangtze River Delta regionwas finally accomplished. The results indicated that:①in winter, in comparison to the atmospheric CO2 concentration simulated based on EDGAR v6.0, the atmospheric CO2 concentration simulated based on the modified inventory was more consistent with observed values. ②The simulated atmospheric CO2 concentration was higher than observation at night and lower than observation during the daytime. The CO2 emission data of emission inventories could not fully reflect the diurnal variation in anthropogenic emissions, andtheoverestimation, caused by the simulated low-atmospheric boundary layer height at night, of the contribution from point sources with higher emission height near the observation station were the main reasons. ③The simulation performance on atmospheric CO2 concentration was greatly affected by the emission bias of the EDGAR grid points that significantly contributed to concentrations of the observation station, and this indicated that the uncertainty in the spatial distribution in EDGAR emission was the main factor influencing the simulation accuracy. ④The posterior anthropogenic CO2 emission flux in the Yangtze River Delta from December 2017 to February 2018 was around (0.184±0.006) mg·(m2·s)-1and (0.183±0.007) mg·(m2·s)-1 based on EDGAR and the modified inventory, respectively. It is suggested that the inventories with higher temporal and spatial resolutions and more accurate spatial emission distribution should be selected as the prior emissions to obtain a more accurate estimation of the regional anthropogenic CO2 emissions.

[Soil Heavy Metal Content, Pollution, and Influencing Factors in Typical Farming Area of Sichuan Basin].

In order to identify heavy metal contents, the pollution characteristics and influencing factors of soil in typical farming areas in the Sichuan basin were analyzed, with Jiangjin District of Chongqing City chosen as the study area. Two hundred and forty-seven topsoil samples were collected and analyzed using the Nemerow index (NPI), geographical information system (GIS), and geodetector method. The results demonstrated that: 1 the arithmetic means of Cd, Cu, and Zn in the topsoil were 1.22, 1.10, and 1.98 times that of the soil background values in western Chongqing, respectively. 2 The high-value areas of the six heavy metals were mainly distributed in the northern, western, and central Jiangjin district, whereas the low-value areas were distributed in the eastern and southern Jiangjin district. 3 The NPI showed that the polluted sample points accounted for 22.1% of the total sample points, indicating that the overall soil pollution was mainly safety and vigilance in general. The high value of NPI was distributed in Dingshan street in the northern Jiangjin district. 4 The explanatory power of stratum on the distribution of heavy metal contents in the topsoil was the strongest, followed by that of the topographic factor. The interaction effect of the stratum and topographic factors on the distribution of heavy metal content in soil was the strongest. The results showed that the stratum and topographic factors were the key factors affecting the distribution of soil heavy metal contents in the study area.

[Effect of electroacupuncture on visceral sensitivity and colonic NGF, TrkA, TRPV1 expression in IBS-D rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) on mental state, visceral sensitivity and protein expression of nerve growth factor (NGF), tyrosine kinase receptor A (TrkA) and transient receptor potential vanilloid 1 (TRPV1) of colonic tissue in diarrhea-predominant irritable bowel syndrome (IBS-D) rats, and to explore its possible mechanism on treating IBS-D. METHODS: A total of 36 male SD rats of SPF grade were randomized into a blank group, a model group, an EA group and a western medication group, 9 rats in each group. In the model group, the EA group and the western medication group, IBS-D model was established by enema of dinitrobenzene sulfonic acid (DNBS) combined with chronic restraint stress method. In the EA group, EA was applied at "Tianshu" (ST 25) and "Shangjuxu" (ST 37), with disperse-dense wave, in frequency of 2 Hz/100 Hz, 20 min each time, once a day for 7 days. In the western medication group, pinaverium bromide suspension was given by gavage (15 mg•kg-1•d-1) for 7 days. Before and after model establishment, and after intervention, the body mass, 24 h food intake and fecal water content were observed, the visceral sensitivity was detected by abdominal withdrawal reflex (AWR); after intervention, the mental state was evaluated by elevated plus maze (EPM) test, the protein expression of NGF, TrkA and TRPV1 was detected by immunohistochemistry and Western blot in the 4 groups. RESULTS: After model establishment, compared with the blank group, the body mass and 24 h food intake were decreased (P<0.05), first systolic latency of AWR was shortened and number of contraction wave of AWR was increased (P<0.05), and fecal water content was increased (P<0.05) in the model group, the EA group and the western medication group. After intervention, compared with the blank group, open arm residence time ratio (OT%) of EPM was decreased (P<0.05) and protein expression of NGF, TrkA, TRPV1 in colonic tissue was increased in the model group (P<0.05); compared with the model group, the body mass and 24 h food intake were increased (P<0.05), first systolic latency of AWR was lengthened and number of contraction wave of AWR was decreased (P<0.05), the fecal water content was decreased (P<0.05), OT% of EPM was increased (P<0.05), and protein expression of NGF, TrkA, TRPV1 in colonic tissue was decreased (P<0.05) in the EA group and the western medication group. CONCLUSION: Electroacupuncture at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) can relieve the anxiety and depression-like behaviors in IBS-D rats, down-regulate the protein expression of NGF, TrkA, TRPV1 in colonic tissue, so as to reduce the visceral sensitivity and relieve symptoms.

[Evaluation of gap-filling methods for CH4 flux data based on eddy covariance method in the Lake Taihu, China]. / æ¶¡åº¦ç›¸å ³è§‚æµ‹çš„å¤ªæ¹–CH4通量数据插补方法评价.

Eddy covariance method has become a key technique to measure CH4 flux continuously in lakes. A large number of CH4 flux data was missing due to variable reasons. In order to reconstruct a complete time series of CH4 flux, it is necessary to find an appropriate gap-filling method to insert the CH4 flux data gap. Based on the routine meteorological data and CH4 flux data measured at Bifenggang site in the eastern part of the Taihu eddy flux network during 2014 to 2017, we analyzed the control factors of CH4 flux at the half-hour scale and daily scale. With those data, we tested that whether nonlinear regression method and two machine learning methods, random forest algorithm and error back propagation algorithm, could fill the CH4 flux gap at the half-hour scale and daily scale. The results showed that CH4 flux at the half-hour scale was mainly influenced by sediment temperature, friction velocity, air temperature, relative humidity, latent heat flux and water temperature at 20 cm in the growing season, and was mainly affected by relative humidity, latent heat flux, wind speed, sensible heat flux and sediment temperature in non-growing season. The CH4 flux at the daily scale was mainly affected by latent heat flux and relative humidity. Random forest model was the best in CH4 flux data gap filling at both time scales. The random forest model with the input variables of day of year, solar elevation angle, sediment temperature, friction velocity, air temperature, water temperature at 20 cm, relative humidity, air pressure, and wind speed was more suitable for filling the CH4 flux data gap at the half-hour scale. The random forest model with the input variables of day of year, sediment temperature, friction velocity, air temperature, water temperature at 20 cm, relative humidity, air pressure, wind speed, and downward shortwave radiation was more suitable for filling CH4 flux data gap at the day scale. The interpolation models could fill the data gap better at daily scale than that at the half-hour scale.

[Temporal and Spatial Variation Characteristics of Methane, Carbon Dioxide, and Nitrous Oxide Concentrations and the Influencing Factors in Small Aquaculture Pond].

As an important source of greenhouse gases, the changes in greenhouse gas concentrations of aquaculture ponds are not only the basis for accurate quantification of greenhouse gases emissions but are also important for identifying their influencing factors. The spatial and temporal variation characteristics of CH4, CO2, and N2O concentrations and the influencing factors in a typical small aquaculture pond in the Yangtze River Delta were analyzed based on the headspace equilibrium-gas chromatograph method. Except in spring, the concentrations of CH4, and N2O appeared high at noon or afternoon and were influenced by water temperature. Impacted by water temperature and aquatic plant photosynthesis, the concentrations of CO2 were high in the morning when photosynthesis was weak. The concentrations of CH4 and CO2 were the highest in autumn and the lowest in winter. The mean concentrations of CH4 in autumn and winter were 176.34 nmol·L-1 and 32.75 nmol·L-1, respectively, which were mainly affected by air temperature, water temperature, and dissolved oxygen. The average CO2 concentrations in autumn and winter were 134.37 µmol·L-1 and 23.10 µmol·L-1, respectively, and were mainly affected by aquatic vegetation photosynthesis and pH. N2O concentration was the highest in summer and the lowest in winter, with mean values of 97.05 nmol·L-1 and 19.41 nmol·L-1, respectively, which were mainly affected by air temperature and water temperature. In terms of the vertical spatial variations of the three greenhouse gases, the concentration of CH4decreased with water depth in summer, and the concentration differences between the surface layer and the bottom and middle layers were 71.28 nmol·L-1 and 42.80 nmol·L-1, respectively. The concentration of CH4 increased with water depth in autumn, and the concentration difference between the bottom layer and surface layer was 163.94 nmol·L-1. The CO2 concentration increased with water depth in summer and autumn. The concentration differences between the bottom and surface concentrations were 18.69 µmol·L-1 and 29.90 µmol·L-1, respectively. N2O concentration showed no obvious change in the vertical direction. For the horizontal variations, the concentrations of CH4, CO2, and N2O in the feeding area in summer and in chicken manure in spring were approximately 1.34-1.98 times and 1.95-2.42 times those in other areas, respectively, and the concentrations of N2O and CO2 in spring and summer were approximately 1.13-1.26 times and 1.39-1.74 times those in other areas.

[Effects of chamber characteristics on CO2 and CH4 flux at the water-air interface measured by the chamber method]. / 箱体特征对箱式法观测水-气界面CO2和CH4通量的影响.

The chamber method is widely used to measure CO2 and CH4 flux in inland water. However, the designs of chamber used in various studies are different and lack unified standards, which would affect the observation results. To clarify the impacts of chamber characteristics, including light transmittance, air pressure difference inside and outside the chamber, and gas mixing degree in the chamber, on CO2 and CH4 flux measurements at the water-air interface, we compared the effects of transparent/opaque chamber, the chamber with/without air pressure equalizing device and fan on CO2 and CH4 flux measurements in the aquaculture pond, based on the multi-channel closed dynamic chamber system. The results showed that, during the daytime in summer, compared with the transparent chamber which could measure the actual CO2 flux, when CO2 was emitted from the pond, the opaque chamber overestimated the CO2 flux by 90%; when CO2 was absorbed by the pond, the opaque chamber underestimated the CO2 flux by 50%. The CH4 diffusion flux measured by the opaque chamber was 40% lower than that measured by the transparent chamber. There was no significant difference between CO2 and CH4 flux measured by the chamber with and without air pressure equalizing device. CO2 flux observed by the chamber without fan had poor representativeness, being 20% higher than that observed by the chamber with fan. Moreover, CH4 flux emitted through different pathways could not be distinguished using the chamber without fan. Therefore, when the chamber method was used to observe the CO2 and CH4 flux at the water-air interface, the chamber shall be transparent and be installed with fan.

Fatigue Crack Growth Behavior of CP-Ti Cruciform Specimens with Mixed Mode I-II Crack under Biaxial Loading.

Investigations on the fatigue crack growth of commercial pure titanium are carried out with cruciform specimens under different biaxial load ratios (λ = 0, 0.5, and 1) and crack inclination angles (ß = 90°, 60°, and 45°) in this paper. Based on the finite element results, the modified solution of stress intensity factors KI and KII for cruciform specimens containing mixed mode I-II crack is obtained by considering crack size, biaxial load ratio, and crack inclination angles. The experimental results show that the maximum tangential stress criterion is fit for the prediction of crack initiation angles for mixed model I-II crack under uniaxial or biaxial loading condition. When the biaxial load ratio increases, the crack propagation angle becomes smaller, and so does the fatigue crack growth rate of mode I crack or mixed mode I-II crack. Based on an equivalent stress intensity factor, a new valid stress intensity factor is proposed to better describe the biaxial fatigue crack growth behavior, which can demonstrate the contribution of mode I and mode II of stress intensity factor.

Green Tides in the Yellow Sea Promoted the Proliferation of Pelagophyte Aureococcus anophagefferens.

Harmful algal blooms formed by fast-growing, ephemeral macroalgae have expanded worldwide, yet there is limited knowledge of their potential ecological consequences. Here, we select intense green tides formed by Ulva prolifera in the Yellow Sea, China, to examine the ecological consequences of these blooms. Using 28-isofucosterol in the surface sediment as a biomarker of green algae, we identified the settlement region of massive floating green algae in the area southeast of the Shandong Peninsula in the southern Yellow Sea. The responses of the phytoplankton assemblage from the deep chlorophyll-a maximum layer were then resolved using high-throughput sequencing. We found striking changes in the phytoplankton community in the settlement region after an intensive green tide in 2016, characterized by a remarkable increase in the abundance of the pelagophyte Aureococcus anophagefferens, the causative species of ecosystem disruptive brown tides. Our study strongly suggests that the occurrence of massive macroalgal blooms may promote blooms of specific groups of microalgae through alteration of the marine environment.

[Acupuncture and moxibustion treatment for dry eye disease: a network Meta-analysis of rando-mized controlled trial].

OBJECTIVE: A network Meta-analysis based on Bayesian theory was used to evaluate efficacy and safety of acupuncture and moxibustion in the treatment of dry eye disease(DED), so as to provide evidence-based research basis for clinical application. METHODS: Randomized controlled trials (RCT) for acupuncture and moxibustion treatment of DED published from the inception of database to November 25, 2020 were searched from PubMed, Embase, Cochrane Library, Web of Science, Sinomed, CNKI, Wanfang and VIP Database. Two reviewers independently screened the literatures, extracted the data. The quality of the included literature was evaluated, and network Meta-analysis was performed by using Stata14.0 and R4.0.3 software. RESULTS: A total of 71 literatures were identified, including 5 536 patients with DED, covering 11 different interventions. Network Meta-analysis showed that acupuncture+traditional Chinese medicine+artificial tears was the best treatment option in terms of the clinical effective rate, breakup time of tear film (BUT), Schirmer I test (SIT) with surface under cumulative ranking area value. Acupuncture+traditional Chinese medicine+artificial tears was better than artificial tears in the clinical effective rate (odds ratio［OR］=12.34, 95% confidence interval［CI］［4.72, 36.89］), BUT(mean differenc［MD］=2.76, 95%CI［0.16, 5.40］), SIT(MD=4.76, 95%CI［1.23, 8.29］). CONCLUSION: Acupuncture and moxibustion in the treatment of DED are generally better than artificial tears, and acupuncture-moxibustion combined with other traditional Chinese medicine therapy has the best effect.

Distribution of Aureococcus anophagefferens in relation to environmental factors and implications for brown tide seed sources in Qinhuangdao coastal waters, China.

Brown tides caused by Aureococcus anophagefferens have occurred along the Qinhuangdao coastline in the Bohai Sea (BS) in recent years. Little is known about the spatio-temporal distribution of A. anophagefferens, particularly its profile distribution and the effects of environmental controls. In this study, four surveys were conducted in Qinhuangdao coastal waters during the brown tide from June to July 2013; another survey was conducted to cover a larger region in the BS in May 2016. Temperature, salinity, nutrients, and chlorophyll a were analyzed; and the density of A. anophagefferens was detected by a sensitive qPCR method. The intensive brown tide only occurred in Qinhuangdao inshore waters at temperatures ranging from 21.5 to 23.2 °C and relatively high salinity (> 29). Redundancy analysis indicated that the low dissolved inorganic nitrogen limited the growth of other pico- and nano-algal species; high dissolved organic nitrogen and low inorganic nutrients were suitable for the development of brown tides in Qinhuangdao coastal waters, which also contained a thermocline during the brown tide. At the early stage of the brown tide, a high abundance of A. anophagefferens appeared at the bottom of offshore waters characterized by low temperature and high salinity. The A. anophagefferens cells were speculated to originate from water mass located in a depression between the central ridge and the Qinhuangdao coastal area. In brief, this study reported the spatio-temporal variation of brown tides based on the abundance of A. anophagefferens and environmental forces and implied that A. anophagefferens could be transported from the bottom of offshore waters to promote brown tides in inshore waters of Qinhuangdao.

[Effect of acupuncture on intestinal flora and TLR4 in brain and intestinal tissues in rats with stress gastric ulcer].

OBJECTIVE: To observe the effect of acupuncture at "Guanyuan" (CV 4) and "Xiajuxu" (ST 39) on intestinal flora and Toll-like receptors-4 (TLR4) in brain and intestinal tissue in rats with stress gastric ulcer (SGU), and to explore the possible mechanism of acupuncture for SGU. METHODS: Thirty-one male SD rats were randomly divided into a blank group (n=7), a model group (n=8), an acupuncture group (n=8) and a drug group (n=8). The rats in the model group, acupuncture group and drug group were treated with modified restraint plus water-immersion stress method to establish SGU model. After modeling, the rats in the acupuncture group were treated with acupuncture at "Guanyuan" (CV 4) and "Xiajuxu" (ST 39), 20 min each time, and the needles were twirled for 30 s every 5 min. The rats in the drug group were treated with intragastric administration of 2 mL omeprazole enteric-coated tablets (20 mg/mL). Both the treatments were given once a day for 5 days. After the intervention, the gastric mucosal damage index was calculated by Guth method; the morphological changes of gastric mucosa were observed by HE staining; the diversity of intestinal flora was detected by 16S rDNA high-throughput sequencing; TLR4 contents in brain and intestinal tissues were determined by ELISA. RESULTS: Compared with the blank group, the gastric mucosal damage index was significantly increased in the model group (P<0.01); the morphological changes of gastric mucosa were obvious; the Observed Species index and Shannon index of α diversity index of intestinal flora were decreased (P<0.05); the ß diversity showed that the spatial distance between the model group and the blank group was far; the TLR4 contents in the brain and intestinal tissue were increased (P<0.05, P<0.01). Compared with the model group, the gastric mucosal damage index was decreased in the acupuncture group and the drug group (P<0.05); the morphology of gastric mucosa was improved; the Observed Species index and Shannon index of α diversity index of intestinal flora in the acupuncture group was increased (P<0.05), and the Shannon index in the drug group was increased (P<0.05); the ß diversity showed that the spatial distance between the acupuncture group and the blank group was close; the TLR4 contents in the brain and intestinal tissues of the acupuncture group were decreased (P<0.01). Compared with the drug group, the contents of TLR4 in the intestinal tissue of the acupuncture group were decreased (P<0.05). CONCLUSION: Acupuncture at "Guanyuan" (CV 4) and "Xiajuxu" (ST 39) could alleviate SGU in rats, and its mechanism may be related to increasing the diversity of intestinal flora, promoting the disorder of intestinal flora to normal, and reducing the overexpression of TLR4 in brain and intestinal tissues.

Targeting the Epithelial-to-Mesenchymal Transition in Cancer Stem Cells for a Better Clinical Outcome of Glioma.

Glioma is one of the most common malignant tumors of the central nervous system with a poor prognosis at present due to lack of effective treatment options. Its initiation, migration, and multipotency are affected by cancer stem cell's transition. Previous studies imply that changes in the cancer stem cells can affect the malignant differentiation of the tumor. We found that the epithelial-to-mesenchymal transition (EMT)-related regulatory pathway is an important target for tumor therapy. In this review, we discuss the transition factor of EMT and 3 specific pathways that affect the EMT of cancer stem cells during tumor development. We conclude that targeting the EMT process of cancer stem cells can be a feasible approach in the treatment of glioma.

Distribution of Alexandrium pacificum cysts in the area adjacent to the Changjiang River estuary, China.

The coastal waters adjacent to the Changjiang River estuary (CRE) are characterized by nutrient pollution and recurrent harmful algal blooms. In this study, resting cysts of Alexandrium pacificum Litaker and A. catenella (Whedon & Kof.) Balech, two major species within the A. tamarense species complex in Chinese coastal waters, were studied using sediment samples collected from the area adjacent to the CRE in May 2014 and December 2015. Cysts were detected with two real-time quantitative PCR assays, as well as the primuline-staining method. Only cysts of A. pacificum were found in the study area, which mainly distributed in the mud depositional zone near the CRE. A low-abundance region of the cysts present in spring is in accordance with the intrusive pathway of the Nearshore Kuroshio Branch Current (NKBC), suggesting that A. pacificum blooms could be regulated by seasonal intrusion of NKBC.

[Effect of acupuncture on intestinal flora in rats with stress gastric ulcer].

OBJECTIVE: To observe the effect of acupuncture at "Baihui" (GV 20), "Zhongwan" (CV 12) and "Zusanli" (ST 36) on intestinal flora in rats with stress gastric ulcer (SGU) , and to explore the mechanism of acupuncture promoting SGU recovery. METHODS: Thirty-one SPF SD rats were randomly divided into a control group (7 rats), a model control group (8 rats), an acupuncture group (8 rats) and a medication group (8 rats). The rats in the model group, acupuncture group and medication group were selected to applied the improved restraint water-immersion stress method to establish the SGU model. After modeling, the rats in the control group and model group were fixed and restrained for 20 min every day for a total of 5 days; the rats in the acupuncture group were intervented with acupuncture at "Baihui" (GV 20), "Zhongwan" (CV 12) and "Zusanli" (ST 36), once a day, 20 min each time, and twisting needle for 30 s every 5 min for a total of 5 days; the rats in the medication group were gavaged by solution of omeprazole enteric-coated tablet (200 mg/mL), 2 mL for each rat, once a day. Guth method was used to calculate the gastric mucosal damage index (GMDI), HE staining was used to observe the pathological changes of gastric mucosa, and 16SrDNA identification was used to detect the structural abundance of intestinal flora. RESULTS: Compared with the control group, the GMDI of rats in the model group was increased (P<0.01), the gastric mucosal pathological changes were significant, and the intestinal flora richness index Chao1, Observed species and diversity index Shannon were all decreased (P<0.05), the diversity index Simpson was increased (P<0.05). Compared with the model group, the GMDI of rats in the acupuncture group and medication group was reduced (P<0.01, P<0.05), the gastric mucosal damage degree was reduced, and the intestinal flora richness index Chao1, Observed species and diversity index Shannon were all increased (P<0.05) and the diversity index Simpson decreased (P<0.05). Compared with the medication group, the GMDI of rats in the acupuncture group was reduced (P<0.01), the recovery of gastric mucosal injury was better than that of the medication group. CONCLUSION: Acupuncture can effectively improve gastric mucosal injury of SGU, and the mechanism may be related to increasing the diversity of intestinal flora and promoting the correction of the disordered intestinal flora.

[Effect of acupuncture on serum inflammatory cytokines and intestinal flora in rats with stress-induced gastric ulcer].

OBJECTIVE: To investigate the effect of acupuncture on serum inflammatory cytokines and intestinal flora in rats with stress-induced gastric ulcer (SGU), and to explore the mechanism of acupuncture in the treatment of SGU. METHODS: Sprague-Dawley rats were randomly divided into blank, model, acupuncture, and medication groups, with 7 rats in each group. Restraint water-immersion stress was used to establish the model of SGU. The rats in the acupuncture group were given acupuncture at "Zhongwan"(CV12) and bilateral "Zusanli"(ST36) for 20 min, once a day for 5 days, and those in the medication group were given 2 mL solution of Omeprazole enteric-coated tablets (0.2 mg/kg) by gavage, once a day for 5 days. The Guth method was used to calculate the gastric mucosa damage index, HE staining was used to observe the histopathological changes of the gastric mucosa, ELISA was used to measure the serum levels of interleukin-4 (IL-4) and interleukin-6 (IL-6), and 16S rDNA sequencing method was used to observe the change in intestinal flora. RESULTS: Compared with the blank group, the model group had a significant increase in gastric mucosa damage index (P<0.01), markedly pathological changes of the gastric mucosa shown by HE staining, a significant reduction in the content of serum IL-4 (P<0.01), and a significant increase in the content of serum IL-6 (P<0.01), as well as a significant reduction in Bacteroidetes and Firmicutes at the phylum level. Compared with the model group, the acupuncture group and the medication group had a significant reduction in gastric mucosa damage index (P<0.01, P<0.05). HE staining showed reduced pathological changes of the gastric mucosa, as well as a significant increase in the content of serum IL-4 (P<0.01, P<0.05) and a significant reduction in the content of serum IL-6 (P<0.01, P<0.05). As for the intestinal flora, there was a significant increase in Bacteroidetes. Compared with the medication group, the gastric mucosa damage index was decreased （P<0.05），the content of serum IL-4 was significantly increased （P<0.05）, while the content of serum IL-6 significantly decreased (P<0.05) in the acupuncture group. CONCLUSION: Acupuncture at CV12 and ST36 can down-regulate the content of serum IL-6, up-regulate the content of serum IL-4, maintain the relative homeostasis of inflammatory cytokines, and regulate the community structure of intestinal flora, and thus help to repair the damage of gastric mucosa.

Resting cysts of Alexandrium catenella and A. pacificum (Dinophyceae) in the Bohai and Yellow Seas, China: Abundance, distribution and implications for toxic algal blooms.

The Alexandrium tamarense species complex consists of 5 closely related species that are important bloom-forming dinoflagellates with a complex life cycle. The formation of resting cyst is a key strategy to resist harsh environmental conditions. In this study, the resting cysts of two major bloom-forming species of the A. tamarense species complex in China, A. catenella (Whedon & Kof.) Balech (previously A. fundyense, or A. tamarense species complex Group I) and A. pacificum Litaker (A. tamarense species complex Group IV), were studied in surface sediment collected from the Bohai Sea (BS) and Yellow Sea (YS) during two cruises conducted in 2012 and 2015. Cyst abundance of the A. tamarense species complex was first quantified by the primuline-staining method, and cysts of the two species were subsequently determined using two real-time quantitative PCR (qPCR) assays. Results showed that resting cysts of the A. tamarense species complex were more abundant in the YS than the BS (mean of 480 and 33 cysts g dry weight, DW-1 of sediment, respectively). Cysts were mainly found in the central portion of the northern YS, the area SE (southeast) of the Shandong peninsula, and the area near the Subei Shoal in the southern YS, where surface sediment had a high percentage of clay and silt (particle size < 63 µm) content. The maximum cyst abundance recorded was 3090 cysts g DW-1 of sediment in 2012 and 3448 cysts g DW-1 in 2015, respectively. Cysts were mainly composed of A. catenella in the YS and the BS, while those of A. pacificum were only detected occasionally at some sampling sites in the YS. Highly abundant resting cysts in surface sediment of the YS may serve as "seed banks" for recurrent toxic blooms of A. catenella and the associated shellfish contamination by paralytic shellfish toxins in the YS.

Emodin attenuates cell injury and inflammation in pancreatic acinar AR42J cells.

The study was intended to investigate the protective effects of emodin against cell injury and inflammation in AR42J cells. We determined trypsin and lipase activity, intracellular ROS and MMP using specific assay kits. The related protein expression and TNF-α and IL-6 in the medium were assayed by Western blot and ELISA kits. Results showed that emodin could protect AR42J cells against cell injury caused by cerulein and lipopolysaccharide which were possibly associated with inhibition of mitochondrial damage, ROS production, and then significantly inhibited ROS-mediated pathway, and ameliorated pancreatic cells injury by depleting the levels of inflammatory cytokines.

Kisspeptin recombinant oral vaccine: A master gene vaccine inhibiting the reproductive physiology and behavior of ram lambs.

The KISS1 gene product, kisspeptin, stimulates gonadotrophic steroid hormone (GNRH) neuronal signaling through the G-protein coupled receptor, kiss1r. Disturbance of this signaling pathway causes hypogonadotropic hypogonadism in mammals. As part of this cutting-edge research project, we analyzed the efficacy of an oral kisspeptin recombinant vaccine on the reproductive physiology and behavior of ram lambs. Ten 56-day old ram lambs were randomly divided into treatment and control groups to receive the experimental recombinant vaccines, C500/pKS-asd or C500/pVAX-asd (aspartate-ß semialdehyde dehydrogenase), respectively. The vaccines were orally administered at day 0, 28 and 56 and blood samples were taken and scrotal circumference data recorded at 14-day intervals (days 0, 14, 28, 42, 56, 70, and 84). At the end of the experimental period, day 98, sexual behaviors were assessed, scrotal circumferences were measured, and blood samples were collected. Testicular samples were also collected after the animals were sacrificed. Anti-kisspeptin antibody and testosterone serum levels were measured by indirect ELISA. Results demonstrated that the levels of anti-kisspeptin antibodies were significantly higher in the treatment group compared to controls (P<0.05, P<0.01 and P<0.001). However, serum testosterone levels were lower in the treatment group (P<0.01). Interestingly, vaccine administration contributed to a significant reduction (P<0.01) in sexual behavior propensity. These results suggest that the kisspeptin recombinant oral vaccine regulates and inhibits the reproductive physiology and behavior of ram lambs.