First report of jujube anthracnose caused by Colletotrichum siamense in China.

Jujube (Zizyphus jujuba Mill.), a native small deciduous tree of China, is widely cultivated in China, Korea, India, Japan, Europe, and the United States (Chen et al. 2020). The fruit have been commonly consumed as healthy food supplements and traditional Chinese medicine for over 2000 years (Li et al. 2007). In August 2019, anthracnose-like leaf spot symptoms were observed on jujube plants in Xiaomenya Village, Jinan City, Shandong Province, China (36°27'39″N, 117°3'13″E), with over 30% leaf disease incidence. The spots were circular, sunken, brown in the center and with dark brown edges. As the spots enlarged and coalesced, it resulted in leaf perforation and early defoliation. Sometimes acervuli were observed on the lesions (Fig. S1a, b). To identify the causal agent, 20 diseased leaves were sampled, the margins of the lesions were cut into pieces (5 × 5 mm), sterilized and cultured following the protocol described previously (Wan et al. 2020) at 25 ℃ for 5 days. Twelve monospore isolates showing identical colony morphology were obtained. Three representative isolates, JNZG11, JNZG311, JNZG313, were used for further study. When grown on PDA the colony color was initially white and then turned pale-gray to gray in 5-day-old cultures. On the reverse, colonies were brown-black with an orange pigmentation near the center. Aerial mycelium was cottony, dense, white to pale-gray. Conidia were hyaline, 1-celled, smooth-walled, subcylindrical, oblong, attenuated with slightly rounded ends, (11.1-) 12.7-13.3 (-17.8) ×(-4.4) 5.2-5.5 (-6.3) µm (n=50). Appressoria were dark-brown, oval or irregular, (7.3-) 8.6-9.2 (-9.8) ×(-5.1) 5.8-6.9 (-7.0) µm (n=50) (Fig. S1c-g). The morphology resembled those of Colletotrichum gloeosporioides species complex (Cannon et al. 2012). For accurate identification, the sequences of the ribosomal internal transcribed spacer (ITS), actin (ACT), ß-tub2 (TUB2), calmodulin (CAL), chitin synthase (CHS-1), and glyceraldehyde-3phosphate dehydrogenase (GAPDH) of the 3 isolates were sequenced (Weir et al. 2012), and deposited into GenBank (Accession Nos. see Table 1). The six loci (ITS, GAPDH, ACT, CHS-1, CAL, and TUB2) were concatenated and the aligned sequences (1904 bp) were 99.7% homologous to ex-type C. siamense ICMP18578. The sequences of 38 Colletotrichum species (44 isolates) were downloaded from GenBank for phylogenetic analyses. In the maximum likelihood phylogenetic tree generated, the highest log likelihood was -8798.90 and the three isolates were all in the C. siamense clade (bootstrap support 94 %) (Fig. S2). To complete Koch's postulates, 60 healthy, mature jujube leaves on 12 branches (5 leaves per branch) (variety 'Zhongqiuhong') were inoculated with 20 µL of spore suspension (106 conidia/mL) or sterile water as a control. The branches were placed in sterile beakers containing a small amount of sterile water sealed with plastic wrap and maintained at 28 °C, 12 h light/dark. Five days after inoculation, all treated leaves showed the typical anthracnose symptom, similar to that observed in the field (Fig. S1h). The same fungus was re-isolated from the margins of the lesions using the aforementioned methods. Whereas no fungus were isolated from the controls. Previously, C. siamense has been reported to infect Z. mauritiana in China (Shu et al. 2020). To our knowledge, this is the first report of C. siamense causing anthracnose on Z. jujuba in China. This finding provides crucial information for the effective management of this disease.

[Distribution of Bacterial Concentration and Viability in Atmospheric Bioaerosols Under Different Weather Conditions in the Coastal Region of Qingdao].

Bacteria are the most abundant microorganisms in atmospheric bioaerosols, widely distributed in the environment. Bioaerosol samples were collected using the FA-1 impact sampler from October 2013 to January 2021 in the coastal city of Qingdao, and samples stained with a BacLightTM bacterial viability kit were used to measure the concentrations of viable and non-viable bacteria with an epifluorescence microscope. The viable and non-viable bacteria in bioaerosols were characterized during different seasons, with particular attention paid to the distribution characteristics of bacteria on foggy, hazy, and dust days. The results showed that the mean concentrations of total bacteria were (1.06±0.68)×105 cells·m-3 in Qingdao during the sampling period, and those of viable and non-viable bacteria were (8.20±4.88)×103 cells·m-3and (9.74±6.72)×104 cells·m-3, respectively. The seasonal concentrations of non-viable bacteria were the highest in spring and winter and the lowest in summer, whereas that of viable bacteria was highest in spring, followed by those in summer and autumn, and the lowest in winter. Atmospheric bacterial concentrations fluctuated with by month, and total bacteria presented a similar variation pattern with that of non-viable bacteria. The monthly average concentration of non-viable bacteria showed the highest value in March during the spring and the lowest in June during the summer in 2021, whereas the highest value for viable bacteria occurred in May during the spring in 2021 and the lowest in December during the winter in 2020. Viable bacteria concentrations were significantly positively correlated with temperature and significantly negatively correlated with NO2, SO2, and CO. Non-viable bacteria were significantly positively correlated with PM. The bacteria in bioaerosol particles showed bimodal, unimodal, and skewed size distributions, varying with season and month. Under different weather conditions, the concentration of non-viable bacteria on dusty days was significantly higher than that on sunny, foggy, or hazy days, but the bacteria viability was as low as 6.85% due to long-distance transport. Anthropogenic pollution resulted in the lowest viability of bacteria at 4.10% on hazy days, whereas the highest viability in bacteria was 16.26% on foggy days due to high humidity. The size distribution of bacteria in bioaerosol particles under different pollution days showed a bimodal distribution, and the peak size depended on the weather type.

Deglycosylation Increases the Aggregation and Angiogenic Properties of Mutant Tissue Inhibitor of Metalloproteinase 3 Protein: Implications for Sorsby Fundus Dystrophy.

Sorsby fundus dystrophy (SFD) is an autosomal dominant macular disorder caused by mutations in tissue Inhibitor of the metalloproteinase-3 (TIMP3) gene with the onset of symptoms including choroidal neovascularization as early as the second decade of life. We have previously reported that wild-type TIMP3 is an endogenous angiogenesis inhibitor that inhibits Vascular Endothelial Growth Factor (VEGF)-mediated signaling in endothelial cells. In contrast, SFD-related S179C-TIMP3 when expressed in endothelial cells, does not have angiogenesis-inhibitory properties. To evaluate if this is a common feature of TIMP3 mutants associated with SFD, we examined and compared endothelial cells expressing S179C, Y191C and S204C TIMP3 mutants for their angiogenesis-inhibitory function. Western blot analysis, zymography and reverse zymography and migration assays were utilized to evaluate TIMP3 protein, Matrix Metalloproteinase (MMP) and MMP inhibitory activity, VEGF signaling and in vitro migration in endothelial cells expressing (VEGF receptor-2 (VEGFR-2) and wild-type TIMP3 or mutant-TIMP3. We demonstrate that mutant S179C, Y191C- and S204C-TIMP3 all show increased glycosylation and multimerization/aggregation of the TIMP3 protein. In addition, endothelial cells expressing TIMP3 mutations show increased angiogenic activities and elevated VEGFR-2. Removal of N-glycosylation by mutation of Asn184, the only potential N-glycosylation site in mutant TIMP3, resulted in increased aggregation of TIMP3, further upregulation of VEGFR-2, VEGF-induced phosphorylation of VEGFR2 and VEGF-mediated migration concomitant with reduced MMP inhibitory activity. These results suggest that even though mutant TIMP3 proteins are more glycosylated, post-translational deglycosylation may play a critical role in the aggregation of mutant TIMP3 and contribute to the pathogenesis of SFD. The identification of factors that might contribute to changes in the glycome of patients with SFD will be useful. Future studies will evaluate whether variations in the glycosylation of mutant TIMP3 proteins are contributing to the severity of the disease.

Extracellular matrix dysfunction in Sorsby patient-derived retinal pigment epithelium.

Sorsby Fundus Dystrophy (SFD) is a rare form of macular degeneration that is clinically similar to age-related macular degeneration (AMD), and a histologic hallmark of SFD is a thick layer of extracellular deposits beneath the retinal pigment epithelium (RPE). Previous studies of SFD patient-induced pluripotent stem cell (iPSC) derived RPE differ as to whether these cultures recapitulate this key clinical feature by forming increased drusenoid deposits. The primary purpose of this study is to examine whether SFD patient-derived iPSC-RPE form basal deposits similar to what is found in affected family member SFD globes and to determine whether SFD iPSC RPE may be more oxidatively stressed. We performed a careful comparison of iPSC RPE from three control individuals, multiple iPSC clones from two SFD patients' iPSC RPE, and post-mortem eyes of affected SFD family members. We also examined the effect of CRISPR-Cas9 gene correction of the S204C TIMP3 mutation on RPE phenotype. Finally, targeted metabolomics with liquid chromatography and mass spectrometry analysis and stable isotope-labeled metabolite analysis were performed to determine whether SFD RPE are more oxidatively stressed. We found that SFD iPSC-RPE formed significantly more sub-RPE deposits (∼6-90 µm in height) compared to control RPE at 8 weeks. These deposits were similar in composition to the thick layer of sub-RPE deposits found in SFD family member globes by immunofluorescence staining and TEM imaging. S204C TIMP3 correction by CRISPR-Cas9 gene editing in SFD iPSC RPE cells resulted in significantly reduced basal laminar and sub-RPE calcium deposits. We detected a ∼18-fold increase in TIMP3 accumulation in the extracellular matrix (ECM) of SFD RPE, and targeted metabolomics showed that intracellular 4-hydroxyproline, a major breakdown product of collagen, is significantly elevated in SFD RPE, suggesting increased ECM turnover. Finally, SFD RPE cells have decreased intracellular reduced glutathione and were found to be more vulnerable to oxidative stress. Our findings suggest that elements of SFD pathology can be demonstrated in culture which may lead to insights into disease mechanisms.

[Concentrations and Patterns of Atmospheric Particulate Nitrogen and Phosphorus During Different Weather Conditions in Qingdao Coastal Region].

Total suspended particulate (TSP) samples were continuously collected in the Qingdao coastal region from March 2018 to October 2019. The concentrations of dissolved inorganic nitrogen (DIN), dissolved inorganic phosphorus (DIP), dissolved total nitrogen (DTN), dissolved total phosphorus (DTP), total nitrogen (TN), and total phosphorus (TP) in the samples were (7.13±6.59) µg·m-3, (17.42±9.88) ng·m-3, (8.34±7.03) µg·m-3, (25.59±13.67) ng·m-3, (10.68±10.59) µg·m-3, and (76.34±51.79) ng·m-3, respectively. The results showed that the concentrations of different nitrogen and phosphorus species in aerosols varied significantly during the sampling period due to the influence of emission intensity, air mass source, and meteorological conditions. The concentrations of DIN, DTN, and TN were the highest in autumn followed by spring, winter, and summer, showing similar seasonal variations. However, TP showed the highest concentration in spring and the lowest in summer. The concentrations of DIN, DTN, and TN on haze days were 4.3, 3.8, and 4.5 times higher than on non-haze reference days, respectively, and DIP, DTP, and TP concentrations were 1.9, 1.9, and 1.2 times higher, respectively. During the heating period, the DIN/DTN ratio on haze days was (92.65±4.09)%, which was 8.87% higher than on non-haze days. During the non-heating period, the DIN/DTN ratio was (80.52±8.42)%, which was 4.83% higher than the reference days. However, the average DTN/TN ratio on haze days was significantly lower than on non-haze days; the average DTN/TN ratio on haze days was (73.41±12.18)% and (80.36±4.72)% during the heating and non-heating period, respectively, and were 13.35% and 5.92% lower, respectively, on non-haze days. The proportion of DIP to DTP on hazy days increased by only 1.47% relative to non-haze days, while the DIP/DTP ratio varied between hazy days. Due to the influence air mass sources, relative humidity, and atmospheric acidification process, the DTP/TP ratio on haze days increased by 10.58% relative to non-haze days. The concentrations of DIN, DTN, and TN in aerosols affected by dust events were 2.5, 2.6, and 2.6 times higher than on non-dust days, while DIP, DTP, and TP were 4.0, 2.8, and 7.2 times higher, respectively.

Cardiac apoptosis caused by elevated cholesterol level in experimental autoimmune myocarditis.

It has been reported that cholesterol-lowing agents can ameliorate severity of myocarditis. However, the beneficial effect of the agents has been claimed to be independent of cholesterol reduction as there is no significant change in the plasma cholesterol level in myocarditis. In the present study, using experimental autoimmune myocarditis (EAM) rats as an animal model, we demonstrated that EAM induced elevation of cholesterol level and impaired cholesterol efflux capacity in the cardiac tissue. Moreover, serum high-density lipoprotein (HDL) content was reduced and HDL function associated protein Paraoxonase 1 (PON1) activity was decreased. Besides, the major structural protein within HDL, Apolipoprotein A1 (ApoA1) expression in the cardiac tissues was significantly reduced while the level of serum ApoA1 was not significantly altered. Importantly, cholesterol depleting agent methyl-ß-cyclodextrin (MßCD) alleviated the development of EAM, as monitored by decreased ratio of heart weight to body weight (HW/BW), decreased infiltration of inflammatory cells and collagen deposition, improved cardiac function, reduced expression of apoptosis-related protein Bax, Fas, FasL and caspase-3 and increased level of anti-apoptotic protein Bcl-2. These results suggest that reduction of cholesterol level in cardiac tissue could suppress EAM-induced cardiac apoptosis through both intrinsic and extrinsic apoptotic pathways.

[Distribution of Total Microbes in Atmospheric Bioaerosols in the Coastal Region of Qingdao].

In order to determine the concentrations and size distributions of total airborne microbes in atmospheric bioaerosols, bioaerosol samples were collected in the coastal region of Qingdao, China, from Sep. 2016 to Jul. 2017 using size-fractionated bioaerosol samplers. The concentrations of total airborne microbes were then measured using an epifluorescence microscope after staining with DAPI (4', 6-diamidino-2-phenylindole). The results showed that the concentrations of total airborne microbes were in the range of 1.86×105-2.54×106 cells·m-3 in Qingdao during the sampling period, with an average of (6.84±4.83)×105 cells·m-3. The concentrations of total airborne microbes showed seasonal variation, with the highest values occurring in spring and winter, and the lowest values in summer and fall. Statistical analysis showed that the concentrations of total airborne microbes in autumn had a significant seasonal difference with spring and summer (P<0.05). The monthly averaged concentrations of total airborne microbes ranged from 2.65×105 cells·m-3 to 1.12×106 cells·m-3, with the highest values in Feb. 2017 and the lowest in Sep. 2016. Total microbe concentrations also varied greatly over a diurnal timescale in fall and winter during the period of 2015 to 2017, but this variation was not statistically significant (P>0.05). Overall, the size distribution of total airborne microbes was skewed, with the highest ratio occurring at >7.0 µm with a ratio of 20.5%-27.3%. The monthly averaged size distribution of total airborne microbes varied between months, showing two main distribution types, specifically bimodal and skewed patterns, respectively. Spearman's rank correlation analysis showed that the AQI and the concentrations of CO, PM2.5, and PM10 had a significant positive correlation with the concentrations of total airborne microbes (P<0.05), while air temperature, wind speed and direction, NO2, SO2, and O3 were not significantly correlated with the airborne microbe concentration during the sampling period (P>0.05). Multiple linear regression showed that 20.6% of the variation in the total microbe concentrations in bioaerosols correlated with relative humidity and PM2.5.

Sorsby Fundus Dystrophy Mutation in Tissue Inhibitor of Metalloproteinase 3 (TIMP3) promotes Choroidal Neovascularization via a Fibroblast Growth Factor-dependent Mechanism.

Choroidal neovascularization (CNV) leads to loss of vision in patients with Sorsby Fundus Dystrophy (SFD), an inherited, macular degenerative disorder, caused by mutations in the Tissue Inhibitor of Metalloproteinase-3 (TIMP3) gene. SFD closely resembles age-related macular degeneration (AMD), which is the leading cause of blindness in the elderly population of the Western hemisphere. Variants in TIMP3 gene have recently been identified in patients with AMD. A majority of patients with AMD also lose vision as a consequence of choroidal neovascularization (CNV). Thus, understanding the molecular mechanisms that contribute to CNV as a consequence of TIMP-3 mutations will provide insight into the pathophysiology in SFD and likely the neovascular component of the more commonly seen AMD. While the role of VEGF in CNV has been studied extensively, it is becoming increasingly clear that other factors likely play a significant role. The objective of this study was to test the hypothesis that basic Fibroblast Growth Factor (bFGF) regulates SFD-related CNV. In this study we demonstrate that mice expressing mutant TIMP3 (Timp3S179C/S179C) showed reduced MMP inhibitory activity with an increase in MMP2 activity and bFGF levels, as well as accentuated CNV leakage when subjected to laser injury. S179C mutant-TIMP3 in retinal pigment epithelial (RPE) cells showed increased secretion of bFGF and conditioned medium from these cells induced increased angiogenesis in endothelial cells. These studies suggest that S179C-TIMP3 may promote angiogenesis and CNV via a FGFR-1-dependent pathway by increasing bFGF release and activity.

[Impact of a Dust Event on the Size Distribution of Metal Elements in Atmospheric Aerosols at a Coastal Region and over the Ocean].

Size-segregated aerosol samples were collected in the Qingdao coastal region and over the East Sea and the Pacific Northwest from March to April in 2016, and the concentration of metal elements (Al, Na, Fe, Mg, K, Cu, Pb, and Cd) were analyzed. The characteristics and the variation in the size distribution of the metal elements in the samples during dusty weather were discussed, as well as the sources and factors that influence the metals. The results indicated that the concentration of Al, Fe, K, Na, Cu, and Pb in the aerosols in the Qingdao coastal samples increased significantly by 85%-1400%, while the concentration of Cd decreased by 8%; the concentrations of Al, Fe, K, Cu, Pb, and Cd in the samples over the ocean increased by 163%-4580%, while the concentration of Na decreased by 62% during dusty weather. In addition, the size distribution of K changed from a double-peak to a coarse peak mode; that of Pb and Cd changed from a fine peak mode to a double-peak distribution. Cu still presented a double-peak distribution in which the proportion of coarse particles increased; Al, Fe, Mg, and Na showed little variation in their size distribution with a single peak in coarse mode. The dust event had different effects on the size distributions of the metal elements in the coastal and marine aerosol samples. The size distribution of the metal elements (Al, Fe, Mg, Na, and K) in the aerosols collected at the Qingdao coastal site had one peak of 2.1-3.3 µm during dusty weather; the peak moved to 3.3-4.7 µm for marine samples. In addition, the proportion of Al, Fe, Mg, Na, K, Pb, and Cd in the coarse mode decreased by 1%-35% for dust aerosol samples collected in Qingdao coastal region, whereas the proportion of these metals in coarse mode increased by 4%-33% for samples collected over the East Sea. This difference may be related to the hygroscopic growth, transport path, and height of the dust.

Equilibrium, kinetics and thermodynamics of Cu(II) biosorption on Chinese chestnut shell pretreated with steam explosion.

The shells of Chinese chestnuts (Castanea mollissima) are an agricultural residue. This work aimed to evaluate this feasibility of using steam explosion to modify this residue for Cu(II) biosorption from aqueous solutions. Equilibrium, kinetic and thermodynamic parameters were evaluated. The steam-explosion pretreatment increased the surface area of the chestnut shell and exposed more hydroxyl and carboxyl groups, which are binding sites for Cu(II). It changed the sorption from a spontaneous process driven by enthalpy to a nonspontaneous one driven by entropy. It increased the Cu(II) sorption capacity at higher temperatures while it decreased the capacity at lower ones. Compared with untreated chestnut shell, the steam-exploded shell is preferable for Cu(II) sorption at higher temperatures.

[Concentrations and Compositions of Different Forms of Nitrogen and Phosphorus in Atmospheric Aerosols in the Qingdao Coastal Region and over the Yellow and Bohai Sea].

The total suspended particulate (TSP) samples were collected in the Qingdao coastal region and over the Yellow and Bohai Sea from June to July in 2016. The diurnal and nightly TSP samples were also continuously collected in the Qingdao coastal region from August 6 to 15. The concentrations of dissolved inorganic nitrogen (DIN), dissolved inorganic phosphorus (DIP), dissolved total nitrogen (DTN), dissolved total phosphorus (DTP), total nitrogen (TN), and total phosphorus (TP) in the TSP samples were analyzed. Results showed that the concentrations of different forms of nitrogen and phosphorus in Qingdao were higher than those over the Yellow and Bohai Sea during the same sampling period. The contribution of dissolved N was similar to that of insoluble N to TN in Qingdao, with the ratio of DTN to TN of 56%. However the DTN was the dominant contributor of TN over the Yellow and Bohai Sea, accounting for 72% of TN on average. The inorganic nitrogen was the dominant species of DTN in Qingdao and over the Yellow and Bohai Sea (YBS), accounting for 67% and 75% of DTN, respectively. The contribution of dissolved P to TP was similar to that of insoluble P to TP in Qingdao and over the Yellow and Bohai Sea, and the DTP accounted for 49% and 58% of TP in Qingdao and over YBS, respectively. The ratio of IP to DTP was slightly higher than that of OP, with values of 56% and 59% in Qingdao and over the YBS, respectively. The origin of the air mass affected the concentrations and compositions of nitrogen and phosphorus in the aerosols. The concentrations of DIN, dissolved organic nitrogen (DON), TN, DIP, and dissolved organic phosphorus (DOP) in the aerosols from southern air mass were higher than those from northern and marine air masses. The concentration of DON in the diurnal aerosol samples was similar to that in the nightly samples; however, the concentrations of DIN and TN were higher in the diurnal aerosols than those in the nightly aerosols. The DTN was the dominant species of TN in the diurnal and nightly aerosol samples, accounting for 79% of TN on average. Inorganic nitrogen was the dominant species of DTN in the diurnal and nightly aerosols. The ratio of DIN to DTN decreased from 70% in diurnal samples to 61% in nightly samples. The concentrations of DIP were close to that of DOP in the diurnal and nightly aerosols; however, the concentrations of TP were higher in the diurnal aerosols than in the nightly ones. The insoluble P was the dominant form of TP in the aerosols, accounting for 83% and 62% of TP during the day and night, respectively. The contribution of DTP to TP in the nightly aerosols samples was much higher than that in the diurnal aerosols. For both day and night samples, inorganic phosphorus was the dominant species of DTP in aerosols, with a ratio of 71%-77%.

[Formation and Size Distribution of the Secondary Aerosol Inorganic Ions in Different Intensity of Haze in Qingdao, China].

Size-segregated atmospheric aerosol samples were collected from September 2015 to February 2016 at a coastal site in Qingdao, and the concentrations of major water-soluble inorganic ions were analyzed by ion chromatography. Characteristics and variation of size distribution of secondary inorganic components in aerosol were discussed, as well as the formation process and influencing factors of SNA(sulfate, nitrate and ammonium). The results indicated that the concentrations of NO3-, SO42-, NH4+, NO2-, Cl- in the aerosols were in the range of 10.32-193.46, 4.42-74.05, 2.21-57.75, 0.05-2.22 and 1.35-17.39 µg·m-3 respectively. And the mass concentration of SNA increased with the intensity of haze pollution. The concentrations of NO3- on the slight, mild, moderate and severe haze days were 55%, 77%, 240% and 537% higher than that on non-haze days respectively, while concentrations of SO42- increased by 4.7%, 35%, 77% and 262% respectively, and concentrations of NH4+ increased by 72%, 83%, 201% and 526% respectively. The contribution of these water-soluble ions to PM2.5 showed that the proportion of sum of NO3-, SO42-, NH4+, NO2- and Cl- to PM2.5 was in range of 62.03%-80.93%. The proportion of ion to PM2.5 decreased in the order of NO3- > SO42- > NH4+ > Cl- > NO2-. With the enhancement of haze pollution, the proportion of NO3- in PM2.5 increased from 29.53% to 45.54%. The correlation analysis showed that NO3- and SO42- in the fine particle were significantly correlated with gaseous precursors NO2 and SO2, and also showed good correlations with relative humidity, visibility, wind speed and other weather conditions. These results indicated that the formation of SNA in fine particles was one of the main reasons for visibility decrease and the formation of air pollution in haze days. Meanwhile, high concentration of gaseous precursors, high relative humidity and low wind speed were the important influencing factors of haze formation. Except for slight haze days, SOR and NOR in the haze days were higher than those on the non-haze days, and increased significantly with the intensifying of haze, especially for 0.43-0.65 and 0.65-1.1 µm particle size. Conversion rates of nitrogen and sulfur in severe haze days were 1.5 times that in non-haze days, which showed nitrate and sulfate in these fine mode were mainly from gas-to-particle conversion. NO3-, SO42-, NH4+ and NO2- increased in haze significantly, which mainly existed in the fine particles. The cloud process played a more important role on haze days. While on non-haze day, cloud process and the heterogeneous reaction were both the main factors. And the highest proportion of fine mode concentration to total one was observed for NO3-(79.4%) and SO42-(74.4%) on severe haze days respectively. NO3- showed a bimodal distribution with peaks in the size-bin of 0.43~0.65 µm and 3.3-4.7 µm on non-haze, slight, mild haze days, and the fine peak moved to 0.65-1.1 µm on moderate haze days, however the bimodal distribution changed to unimodal distribution with peak at 0.65-1.1 µm on severe hazy days. SO42- showed a bimodal distribution with peaks at 0.43-0.65 µm and 2.1-3.3 µm in the non-haze weather, while the size distribution changed to unimodal distribution on hazy days. But the peak sizes were different in different intensity of haze, with peak at 0.43-0.65 µm on mild and slight haze and 0.65-1.1 µm in moderate and severe haze days. NH4+ showed a single peak distribution in the fine mode, with the peak in the particle size of 0.43-0.65 µm on slight and non-hazy days, and 0.65-1.1 µm on mild, moderate and severe hazy days. Therefore, haze has a great influence on the size distribution of SNA.

[Influence of Dust Events on the Concentration and Size Distribution of Microorganisms in Bioaerosols].

In order to study the influence of dust events on the airborne microbes in atmospheric bioaerosols, bioaerosol samples were continuously collected from March to April 2015 during two dust events using size-fractionated bioaerosol samplers in Lanzhou and Qingdao. The concentration of airborne microbes were measured using an epifluorescence microscope after staining with DAPI (4',6-diamidino-2-phenylindole). The results showed that the concentration of airborne microbes increased significantly during the period of dust (P<0.05). The average concentrations of airborne microbes on sunny days were 5.61×105 cells·m-3 and 2.08×105 cells·m-3 in Lanzhou and Qingdao, respectively, whereas the mean concentrations on dusty days were 14.8 times and 6.42 times those on sunny days, respectively. The concentration of airborne microbes presented a bimodal size distribution on sunny days in Lanzhou and Qingdao, with a peak at>7.0 µm and the lowest values ranging from 4.7 µm to 7.0 µm. However, the second peak appeared at 3.3-4.7 µm and 1.1-2.1 µm in Lanzhou and Qingdao, respectively. The size distribution of airborne microbes changed during the dust period. The size distribution of microbes still presented a bimodal distribution in Lanzhou; however, one peak shifted from>7.0 µm to 1.1-2.1 µm. The size distribution in Qingdao changed from a bimodal distribution to a skewed distribution with the predominant fraction in coarse mode. The background values of microbial load in PM10 before the dust were 2224 cells·µg-1 and 1550 cells·µg-1 in Lanzhou and Qingdao, respectively, and the highest value of such increased significantly to 26442 cells·µg-1 and 10250 cells·µg-1 during the dust events, respectively. This demonstrated that the high concentration of microbes in the dust events resulted not only from increased atmospheric particles but also from dust source and transportation path along with long-range transported dust particles.

Mixture of Peanut Skin Extract and Fish Oil Improves Memory in Mice via Modulation of Anti-Oxidative Stress and Regulation of BDNF/ERK/CREB Signaling Pathways.

Long-term use of fish oil (FO) is known to induce oxidative stress and increase the risk of Alzheimer's disease in humans. In the present study, peanut skin extract (PSE), which has strong antioxidant capacity, was mixed with FO to reduce its side effects while maintaining its beneficial properties. Twelve-week Institute of Cancer Research (ICR) mice were used to conduct animal behavior tests in order to evaluate the memory-enhancing ability of the mixture of peanut skin extract and fish oil (MPF). MPF significantly increased alternations in the Y-maze and cognitive index in the novel object recognition test. MPF also improved performance in the water maze test. We further sought to understand the mechanisms underlying these effects. A significant decrease in superoxide dismutase (SOD) activity and an increase in malonyldialdehyde (MDA) in plasma were observed in the FO group. The MPF group showed reduced MDA level and increased SOD activity in the plasma, cortex and hippocampus. Furthermore, the gene expression levels of brain-derived neurotrophic factor (BDNF) and cAMP responsive element-binding protein (CREB) in the hippocampus were increased in the MPF group, while phosphorylation of protein kinase B (AKT), extracellular signal-regulated kinase (ERK) and CREB in the hippocampus were enhanced. MPF improves memory in mice via modulation of anti-oxidative stress and activation of BDNF/ERK/CREB signaling pathways.

Insolubilization of Chestnut Shell Pigment for Cu(II) Adsorption from Water.

Chestnut shell pigment (CSP) is melanin from an agricultural waste. It has potential as an adsorbent for wastewater treatment but cannot be used in its original state because of its solubility in water. We developed a new method to convert CSP to insolubilized chestnut shell pigment (ICSP) by heating, and the Cu(II) adsorption performance of ICSP was evaluated. The conversion was characterized, and the thermal treatment caused dehydration and loss of carboxyl groups and aliphatic structures in CSP. The kinetic adsorption behavior obeyed the pseudo-second-order rate law, and the equilibrium adsorption data were well described with both the Langmuir and the Freundlich isotherms. ICSP can be used as a renewable, readily-available, easily-producible, environmentally-friendly, inexpensive and effective adsorbent to remove heavy-metal from aquatic environments.

Comparison of Antioxidant Activities of Melanin Fractions from Chestnut Shell.

Chestnut shell melanin can be used as a colorant and antioxidant, and fractionated into three fractions (Fr. 1, Fr. 2, and Fr. 3) with different physicochemical properties. Antioxidant activities of the fractions were comparatively evaluated for the first time. The fractions exhibited different antioxidative potential in different evaluation systems. Fr. 1, which is only soluble in alkaline water, had the strongest peroxidation inhibition and superoxide anion scavenging activity; Fr. 2, which is soluble in alkaline water and hydrophilic organic solvents but insoluble in neutral and acidic water, had the greatest power to chelate ferrous ions; and Fr. 3, which is soluble both in hydrophilic organic solvents and in water at any pH conditions, had the greatest hydroxyl (·OH) and 1,1-diphenyl-2-picryl-hydrazyl (DPPH·) radicals scavenging abilities, reducing power, and phenolic content. The pigment fractions were superior to butylated hydroxytolune (BHT) in ·OH and DPPH· scavenging and to ethylene diamine tetraacetic acid (EDTA) in the Fe(2+)-chelation. They were inferior to BHT in peroxidation inhibition and O2·(-) scavenging and reducing power. However, BHT is a synthetic antioxidant and cannot play the colorant role. The melanin fractions might be used as effective biological antioxidant colorants.

[Microbial Activity in Bioaerosols in Winter at the Coastal Region of Qingdao].

In order to study the level and size distribution of microbial activity of atmospheric bioaerosols, bioaerosol samples were collected from November 2015 to January 2016 using size-fractionated bioaerosol samplers at the coastal region of Qingdao, and the microbial activity was measured using the fluorescein diacetate (FDA) hydrolysis method. The results showed that the level of microbial activity was in the range of 21.89-108.59 ng·m-3 sodium fluorescein during the sampling period, with an average of 59.43 ng·m-3 sodium fluorescein in Qingdao. Size distribution of microbial activity exhibited a tendency, the activity increased with increasing particle size. The microbial activity on particles with coarse size (>2.1 µm) was higher than that on fine size, with the highest average proportion of 24.06% for coarse size larger than 7.0 µm. The daily variation of microbial activity was different for different samples, which showed no significant diurnal variation in winter. The correlation analysis showed that microbial activity was significantly correlated with wind velocity(r=0.445, n=33, **P<0.01) during the sampling period. However, microbial activity showed no significant correlation with the meteorological factors, such as temperature,relative humidity and UV intensity during the sampling period. Moreover, there was no significant correlation of microbial activity with air quality factors, such as AQI, PM2.5, PM10, CO, NO2, O3 and SO2. Source of air mass had significant impact on microbial activity. The average level of microbial activity was 100.33 ng·m-3 sodium fluorescein on sunny days, and the level decreased to 56.53 ng·m-3 sodium fluorescein on hazy days. When the haze was mixed with fog and this special circumstance lasted for several days, the microbial activity reduced to 37.7% of the level of sunny days. Therefore, consecutive hazy weather had great influence on microbial activity.

Tissue inhibitor of metalloproteinase-3 (TIMP3) promotes endothelial apoptosis via a caspase-independent mechanism.

Tissue inhibitor of metalloproteinases-3 (TIMP3) is a tumor suppressor and a potent inhibitor of angiogenesis. TIMP3 exerts its anti-angiogenic effect via a direct interaction with vascular endothelial growth factor (VEGF) receptor-2 (KDR) and inhibition of proliferation, migration and tube formation of endothelial cells (ECs). TIMP3 has also been shown to induce apoptosis in some cancer cells and vascular smooth muscle cells via MMP inhibition and caspase-dependent mechanisms. In this study, we examined the molecular mechanisms of TIMP3-mediated apoptosis in endothelial cells. We have previously demonstrated that mice developed smaller tumors with decreased vascularity when injected with breast carcinoma cells overexpressing TIMP3, than with control breast carcinoma cells. TIMP3 overexpression resulted in increased apoptosis in human breast carcinoma (MDA-MB435) in vivo but not in vitro. However, TIMP3 could induce apoptosis in ECs in vitro. The apoptotic activity of TIMP3 in ECs appears to be independent of MMP inhibitory activity. Furthermore, the equivalent expression of functional TIMP3 promoted apoptosis and caspase activation in ECs expressing KDR (PAE/KDR), but not in ECs expressing PDGF beta-receptor (PAE/ß-R). Surprisingly, the apoptotic activity of TIMP3 appears to be independent of caspases. TIMP3 inhibited matrix-induced focal adhesion kinase (FAK) tyrosine phosphorylation and association with paxillin and disrupted the incorporation of ß3 integrin, FAK and paxillin into focal adhesion contacts on the matrix, which were not affected by caspase inhibitors. Thus, TIMP3 may induce apoptosis in ECs by triggering a caspase-independent cell death pathway and targeting a FAK-dependent survival pathway.

[Concentration and community diversity of microbes in bioaerosols in the Qingdao coastal region].

Bioaerosol samples were collected in Qingdao coastal region during July 2009 - June 2010 to investigate the concentration and community diversity of microbes in bioaerosols. Microbe concentrations (bacteria and fungi) in marine and terrestrial bioaerosols were determined and diversity indices including Shannon-Weiner index, Simpson's index and Pielou index were calculated in this study. Monthly average concentrations of terrestrial bacteria, marine bacteria, terrestrial fungi and marine fungi were in the ranges of 12-436 CFU x m(-3), 25-561 CFU x m(-3), 0-817 CFU x m(-3) and 11-1346 CFU x m(-3), respectively. There were consistent seasonal variations of these four types of microbe, with higher concentrations in spring and summer and lowest during winter, especially in February. Compared to terrestrial microbes, marine microbes account for higher proportion to the total culturable microbes, with a percentage of 63%. The number of microbial species varied from 17 to 102, and was partially correlated with microbial concentrations, however, it did not show obvious seasonal variation. Based on the analysis of calculated diversity indices, we found that the community diversities of four types of microbe were much higher in January, November and May than in February. The community diversity varied with the season, space and different microbial species, and showed a different seasonal variation from the microbial concentration.

Structure-activity relationship of Baifuzi-cerebrosides on BKCa channel activation.

Our previous study reported that a mixture of cerebrosides from traditional Chinese medicine Baifuzi could activate BKCa channel. It is curious to know the effect of each single cerebroside on the channel. Here we isolated 5 pure cerebrosides from the mixture and determined their chemical structures. The most potent one increased the single channel open probability 6 folds with EC50 value of 1.0 µM. The structure-activity relationship revealed that acyl chain length of cerebrosides has potent effect, while configuration of double bond at C8-C9 on their long chain base has weak effect on the channel activity. Thus, this research provides a guide for design and synthesis of a lead cerebroside with more potent effect on the BKCa channel.