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Based on the data o£ daily snowfall and weather phenomena of 11 national meteorological stations in Ulanqab City from 1991 to 2020, the spatial and temporal distribution characteristics of snowstorm were analyzed. The results show that the snowstorm in Ulanqab had obvious seasonal distribution characteristics, mainly happening in spring (March - May) and autumn (September - November). It also had obvious regional distribution in space, and the snowstorm center appeared in Chahar Right Wing Middle Banner and lining District, namely the east side of the Yinshan Mountain. In the past 30 years, the amount of snowstorm in the whole year in Ulanqab showed a certain fluctuation trend, and the number of snowstorm days had shown an obvious upward trend since 2011. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the expression and functional role of miR143HG in non-small cell lung cancer (NSCLC), and its effect on human lung adenocarcinoma cell behavior. Methods: Differential expression of miR143HG between NSCLC tissues and healthy counterparts was identified through bioinformatic analysis. Subsequently, this expression difference in A549 and BEAS- 2B cells was validated using quantitative polymerase chain reaction (qPCR). Overexpression of miR143HG in A549 cells was achieved through liposome-mediated transfection, while cell proliferation as well as apoptosis levels were assessed using cell counting kit-8 (CCK-8) assay, flow cytometry, and protein blotting. Expression of miR-504 (predicted as a target of miR143HG) was determined. Furthermore, A549 cells that overexpress both miR143HG and miR-504 were generated for comparison with cells overexpressing only miR143HG. Results: Compared to BEAS-2B cells, A549 cells showed significantly reduced miR143HG and elevated miR-504 levels (p < 0.05). In A549 cells with miR143HG overexpression, cell proliferation and miR-504 expression significantly reduced while pro-apoptotic proteins (BAX, p53) significantly increased (p < 0.05). Simultaneous overexpression of miR143HG and miR-504 in A549 cells counteracted the effect of miR143HG alone, thus enhancing proliferation and diminishing pro-apoptotic protein expression, similar to control. Conclusion: Downregulation of MiR143HG occurs in human lung adenocarcinoma cells. Upregulation of MiR143HG impedes A549 cell proliferation, promotes apoptosis through pro-apoptotic protein modulation and suppresses miR-504. Co-overexpression of miR143HG and miR-504 reverses these effects, resembling control conditions. Thus, miR143HG exerts its anti-cancer effect in A549 cells by modulating miR-504 and activating p53 pathway. [ABSTRACT FROM AUTHOR]

The combined treatment method integrated photothermal therapy (PTT) with chemotherapy is extremely promising owing to the synergistic therapeutic effect as compared to single PTT or chemotherapy. To facilitate more novel and facile photothermal-chemotherapy drugs as well as promote controllable combination therapy, we have developed a mild and facile method to fabricate polymer polypyrrole (PPy)-doxorubicin (DOX) nanoparticles (NPs) as pH-responsive drug nanocarriers for synergistic photothermal-chemotherapy. In the nanoplatform, poly-L-lysine (PLL)-modified PPy serves as the photothermal material, and (DOX) molecules are adopted as the chemotherapy agent. Based on the cross-linking reaction of glutaraldehyde, DOX molecules are flexibly and efficiently assembled on the surface of PLL-modified PPy NPs. The obtained PPy-DOX NPs possess high photothermal effect, superior loading capacity of DOX, and controlled drug release behavior. The combination photothermal-chemotherapy based on PPy-DOX NPs has significantly enhanced the antitumor therapy effect. In general, the designed PPy-DOX NPs may be a potential drug delivery nanoplatform for cancer combination therapy. [ABSTRACT FROM AUTHOR]

[Objectives] To identify the species of Hirudo from different sources. [Methods] In accordance with morphological, anatomical and molecular taxonomic characteristics, the identification conclusion was drawn. [Results] The living samples of Hirudo belong to 1 order, 2 families, 3 genera and 5 species, and the dry samples of medicinal leeches belong to 4 species. [Conclusions] The sources of medicinal leeches are complex, and there is a problem that Hirudo from different basic sources are mixed for medication. [ABSTRACT FROM AUTHOR]

Objectives] The paper was to establish the quality standard of Shenqi Qiangjing granules and to conduct routine inspection of granular dosage forms. [Methods] The particle size, moisture, loss on drying, dissolubility, content uniformity and microbial limit of Shenqi Qiangjing granules were detected. Radix Astragali, Herba Epimedii, Radix Dipsaci and Fructus Schisandrae Chinensis were qualitatively identified by thin layer chromatography (TLC). [Results] The particle size, moisture, loss on drying, dissolubility, content uniformity and microbial limit of Shenqi Qiangjing granules were all in line with the relevant regulations of Pharmacopoeia of the People's Republic of China (2020 edition). Radix Astragali, Herba Epimedii, Radix Dipsaci and Fructus Schisandrae Chinensis had clear fluorescent spots in TLC assay, and no such spots were found in negative control. [Conclusions] The method has the advantages of simple operation, accurate and reliable results, strong specificity and good repeatability, and can effectively control the quality of Shenqi Qiangjing granules. [ABSTRACT FROM AUTHOR]

In this paper, the resource distribution, pharmacological action and culture status of hirudos were reviewed. Hirudo resources are widely distributed and have a variety of pharmacological effects, but culture is difficult, so the scientific development of hirudos culture and solving the problem of hirudos breeding can alleviate the contradiction in the medicine market and contribute to the sustainable development of hirudo medicine. [ABSTRACT FROM AUTHOR]

[Objectives] To explore the effects of temperature on the primary and secondary metabolism of Scutellaria baicalensis Georgi during the seed germination. [Methods] The superoxide dismutase（SOD） activity was determined using riboflavin-NBT; peroxidase（POD） activity was determined using guaiacol colorimetric method, catalase（CAT）, ascorbate peroxidase（APX）, phenylalanine ammonia lyase（PAL） and cinnamic acid-4-hydroxylase（C4H） activity were detected by ultraviolet spectrophotometry, and chalcone synthase（CHS） activity and the content of secondary metabolites were measured by high performance liquid chromatography（HPLC）. [Results] The germination rate, germination potential and germination index of S. baicalensis seeds were significantly affected by temperature. The most suitable temperature for the germination of S. baicalensis seeds was 25 ℃. The activities of SOD, POD and CAT in S. baicalensis seeds treated at low and high temperature were higher than that treated at suitable temperature; the activities of PAL, C4H and CHS of S. baicalensis seeds treated at low and high temperature were lower than that treated at suitable temperature. There was a good positive correlation between flavonoids and soluble sugar, PAL activity and C4H activity, and the correlation coefficients were R=0.894*, R=0.956* and R=0.951*, respectively. [Conclusions] In adverse environment, S. baicalensis seeds have good defense capabilities. During the germination of seeds, the formation of secondary metabolites is significantly correlated to the activity of key enzymes. Therefore, high-quality medicinal materials can be obtained by taking measures to improve the activity of key enzymes. [ABSTRACT FROM AUTHOR]

Erythropoietin (EPO) drives erythropoiesis and is secreted mainly by the kidney upon hypoxic or anemic stress. The paucity of EPO production in renal EPO-producing cells (REPs) causes renal anemia, one of the most common complications of chronic nephropathies. Although mitochondrial dysfunction is commonly observed in several renal and hematopoietic disorders, the mechanism by which mitochondrial quality control impacts renal anemia remains elusive. In this study, we showed that FUNDC1, a mitophagy receptor, plays a critical role in EPO-driven erythropoiesis induced by stresses. Mechanistically, EPO production is impaired in REPs in Fundc1-/- mice upon stresses, and the impairment is caused by the accumulation of damaged mitochondria, which consequently leads to the elevation of the reactive oxygen species (ROS) level and triggers inflammatory responses by up-regulating proinflammatory cytokines. These inflammatory factors promote the myofibroblastic transformation of REPs, resulting in the reduction of EPO production. We therefore provide a link between aberrant mitophagy and deficient EPO generation in renal anemia. Our results also suggest that the mitochondrial quality control safeguards REPs under stresses, which may serve as a potential therapeutic strategy for the treatment of renal anemia. [ABSTRACT FROM AUTHOR]

[Objectives] The content of total anthraquinone and 5 free anthraquinones in different processed products of rhubarb were determined. [Methods] Using emodin as the reference, the concentration of 1% magnesium acetate methanol solution was used for color development, and the absorbance was measured at 516 nm wavelength; the mobile phase was methanol-0.1% phosphoric acid (76:24) aqueous solution, the detection wavelength was set at 256 nm, the flow rate was 1.0 mL/min, the column temperature was 30 °C, and the sample size was 10 µL. [Results] The content of total anthraquinones in rhubarb was determined by UV spectrophotometry. The linear range was 0.98-23.52 µg/mL, R²=0.999 3, the average recoveries were 100.12%, 99.66% and 99.15%, with RSDs of 1.58%, 0.94% and 1.41%, respectively. The linear ranges of emodin, emodin, emodin, chrysophanol and emodin methyl ether were 0.020 7-0.414(R²=0.999 9), 0.172 1-3.442(R²=0.999 6), 0.020 3-0.406(R²=0.999 9), 0.148-2.96(R²=0.999 8) and 0.164-3.28(R²=0.999 3), respectively. [Conclusions] The contents of total anthraquinones and free anthraquinones in different processed products of rhubarb were determined by UV spectrophotometry and HPLC. The method can be used for quality control of rhubarb and its different processed products. [ABSTRACT FROM AUTHOR]

Thic article reviews the species, identification, chemical composition and pharmacological effects oO hirudo. Hirudo ic widely sourced, difficult te identify, contains a variete of active ingredients, and has a variete of pharmacologicaO effects. In-depth research on hirudo ic of great sienificanco te the qualita control and clinical application of hirudo. [ABSTRACT FROM AUTHOR]

This study was conducted to clarify the roles of macronutrients in regulating plant primary and secondary metabolism of Scutellaria baicalensis Georgi. #Methods] The contents of chlorophyll (Chl) and soluble sugar were detected with ultraviolet spectrophotometry. The activities of phenylalanine ammonia-lyase (PAL) , cinnamate 4-hydroxylae (C4H) and chalcone synthase (CHS) were determined with ultraviolet spectrophotometry. Secondary metabolites were detected by high performance liquid chromatography (HPLC). # Results ] The content of chlorophyll in treatments N, P and K was increased, showing significant diference from that of the control (P < 0. 05) . Among them, Treatment N has the highest content of chlorophyll. Soluble sugar content in treatments N , P and K (K>N>P> CK) was increased considerably and had significant difference from that of the control (P < 0. 05). Compared with the control, PAL and C4 H ac-tivity in treatments N, P and K were increased dramatically by 186. 57% , 134. 09% , 306.91% and 73. 21% , 28. 91% , 247. 57% , respectively. CHS activity was also increased sharply in treatments N, P and K. Scutellarin, baicalin, wogonoside, baicalein, wogonin and oroxylin A contents in all treatments N, P and K (K > N >P) were increased, and had significant difference from that of the control (P<0.05). # Conclusions] The results showed that macronutrients can increase leaf chlorophyll content, soluble sugar content, PAL activity, C4H activity and secondary metabolites of S. baicalensis. [ABSTRACT FROM AUTHOR]

[Objectives] This study was conducted to study the Germination characteristics and secondary metabolism regulation of Scutellaria baicalensis Georgi under osmotic stress. [Methods] The activity of praline, soluble sugar and protein was detected with ultraviolet spectrophotometry. The activity of phenylalanine ammonia-lyase (PAL) and cinnamic acid 4-hydroxylase (C4H) was determined with ultraviolet spectrophotometry. The secondary metabolites were detected by high performance liquid chromatography (HPLC). [Results] The germination percentage, germination potentiality and germination index decreased at first and increased subsequently along with the enhancement of PEG concentration. The praline content, the soluble sugar content and the soluble protein content in S. baicalensis seedlings increased along with the enhancement of PEG concentration and the differences from the CK were significant (P < 0.05). The PAL and C4H in S. baicalensis seedlings decreased after osmotic stress and the differences from the CK were significant (P < 0.05). The results indicate that, the scutellarin, baicalin, wogonoside, baicalein, wogonin and oroxylinA contents decreased after osmotic stress and the differences from the CK were significant (P<0.05). [Conclusions] The quality of the skullcaps could be improved by improving the activity of PAL and C4H. [ABSTRACT FROM AUTHOR]

Bauhinia is a widely planted urban tree plant in tropical cities, which are frequently dwelling in a habitat exposed to inevitable streetlamp lighting. It is of practical meaning to figure out the lighting spectra that benefit nutrient utilization in corporation with a proper exponential fertilization regime. In this study, Bauhinia brachycarpa and B. variegata seedlings were cultured with exponential fertilization at the rate of 80 mg nitrogen (N) plant-1 (N-phosphorus-potassium, 10-7-9) with an unfertilized control, and both were exposed to lightemitting diode spectra of R1BG5 (13.9% red, 77.0% green, 9.2% blue), R2BG3 (26.2% red, 70.2% green, 3.5% blue), and R3BG1 (42.3% red, 57.3% green, 0.4% blue). The R3BG1 spectrum resulted in smaller growing size and dry mass but higher nutrient concentration and root water content compared to the other two spectra. Exponential fertilization increased fresh mass for two Bauhinia species but only increased dry mass in B. brachycarpa. Compared to B. variegata, B. brachycarpa was verified to have a higher capacity to accommodate exogeneous nutrient input through exponential fertilization. The R3BG1 spectrum was recommended for the illumination in streetlamps for Bauhinia because this spectrum can promote nutrient uptake without too fast rate of growth relative to other spectra with lower red-light proportions. [ABSTRACT FROM AUTHOR]

[Objective] This study aimed to investigate the primary and secondary metabolism of Scutellaria baicalensis Georgi during seed germination process under different illumination time. [Method] Chlorophyll (CHL) content, soluble sugar content, phenylalanine ammonia lyase (PAL) activity and cinnamate-4-hydroxylase (C4H) activity were determined with ultraviolet spectrophotometry. The secondary metabolites were detected by high performance liquid chromatography (HPLC). [Result] The results indicated that the germination of S. baicalensis seeds was not sensitive to light and the seedlings were very sensitive to light. CHL content, soluble sugar content, PAL activity and C4H activity increased continuously with the illumination time. The secondary metabolites showed a similar trend. [Conclusion] Illumination time promoted the formation of leaf photosynthetic pigments, thereby affecting the synthesis of primary and secondary metabolites and the activities of PAL and C4H. Therefore, the quality of S. baicalensis can be improved by regulating the illumination time appropriately. [ABSTRACT FROM AUTHOR]

Fluorescence anisotropy (FA) is a reliable, sensitive, and robust assay approach for determination of many biological targets. However, it is generally not applicable for the assay of small molecules because their molecular masses are relatively too small to produce observable FA value changes. To address this issue, we report herein the development of a FA signal amplification strategy by employing graphene oxide (GO) as the signal amplifier. Because of the extraordinarily larger volume of GO, the fluorophore exhibits very high polarization when bound to GO. Conversely, low polarization is observed when the fluorophore is dissociated from the GO. As proof-of-principle, the approach was applied to FA detection of adenosine triphosphate (ATP) with a fluorescent aptamer. The aptamer exhibits very high polarization when bound to GO, while the FA is greatly reduced when the aptamer complexes with ATP, which exhibits a maximum signal change of 0.316 and a low detection limit of 100 nM ATP in buffer solution. Successful application of this strategy has been demonstrated that it can be constructed either in a "signal-off" or in a "signal-on" detection scheme. Moreover, because FA is less affected by environmental interferences, FA measurements could be conveniently used to directly detect as low as 1.0 μM adenosine triphosphate (ATP) in human serum. The universality of the approach could be achieved to detect an array of biological analytes when complemented with the use of functional DNA structures. [ABSTRACT FROM AUTHOR]

Sacbrood virus (SBV) is a picorna-like virus that affects honey bees (Apis mellifera) and results in the death of the larvae. Several procedures are available to detect Chinese SBV (CSBV) in clinical samples, but not to estimate the level of CSBV infection. The aim of this study was develop an assay for rapid detection and quantification of this virus. Primers and probes were designed that were specific for CSBV structural protein genes. A TaqMan minor groove binder (MGB) probe-based, fluorescence real-time quantitative PCR was established. The specificity, sensitivity and stability of the assay were assessed; specificity was high and there were no cross-reactivity with healthy larvae or other bee viruses. The assay was applied to detect CSBV in 37 clinical samples and its efficiency was compared with clinical diagnosis, electron microscopy observation, and conventional RT-PCR. The TaqMan MGB-based probe fluorescence real-time quantitative PCR for CSBV was more sensitive than other methods tested. This assay was a reliable, fast, and sensitive method that was used successfully to detect CSBV in clinical samples. The technology can provide a useful tool for rapid detection of CSBV. This study has established a useful protocol for CSBV testing, epidemiological investigation, and development of animal models. [ABSTRACT FROM AUTHOR]

β-1,3-Dicarbonyl aldehydes were synthesized by iron-catalyzed oxidative reactions between 1,3-dicarbonyl compounds and two molecules of tertiary amines in the presence of tert-butyl hydroperoxide (TBHP). α,β-Unsaturated aldehydes generated by tertiary amine oxidation in situ act as key intermediates under mild reaction conditions. [ABSTRACT FROM AUTHOR]

The cavitation damage of the pollutant wall caused by shock wave and microjet in the process of bubble collapse has attracted widespread attention. However, many researches focus on bubble collapse from the macroscopic experimental point of view, the dynamics of which is still not clear. In this paper, it takes the pressure on the upper and lower wall of pollutants as the research target, and summarizes the influence of initial radius of bubbles, compressive strain rates and temperatures on the release pressure of bubble collapse on the pollutants. The results show that: as the initial radius of bubbles increase, the maximum pressure on the upper and lower wall of benzamide increases first and then decreases. The pressure release rate of bubble increases with the increase of the compressive strain rate. Based on the pressure on the upper and lower wall of the benzamide, the temperature and initial radius of the bubble have a large influence, and the influence of the compressive strain rate is small. The paper provides theoretical guidance for the application of cavitation [ABSTRACT FROM AUTHOR]

Influenza A/H9 viruses circulate worldwide in wild and domestic avian species, continuing to evolve and posing a zoonotic risk. A substantial increase in human infections with A/H9N2 subtype avian influenza viruses (AIVs) and the emergence of novel reassortants carrying A/H9N2-origin internal genes has occurred in recent years. Different names have been used to describe the circulating and emerging A/H9 lineages. To address this issue, an international group of experts from animal and public health laboratories, endorsed by the WOAH/FAO Network of Expertise on Animal Influenza, has created a practical lineage classification and nomenclature system based on the analysis of 10,638 hemagglutinin sequences from A/H9 AIVs sampled worldwide. This system incorporates phylogenetic relationships and epidemiologic characteristics designed to trace emerging and circulating lineages and clades. To aid in lineage and clade assignment, an online tool has been created. This proposed classification enables rapid comprehension of the global spread and evolution of A/H9 AIVs. [ABSTRACT FROM AUTHOR]

[Objectives] Using wild-type AB strain of zebrafish as experimental animal, this study investigated the damaging effect of Wufang Babu Ointment on skin cells, in order to evaluate the skin toxicity of Wufang Babu Ointment. [Methods] Wild-type AB strain of zebrafish with an age of 2 d were taken and fed in different concentrations of Wufang Babu Ointment solution for 24 h. The number of deaths in each group of zebrafish was recorded, and the mortality rate was calculated. Using Origin 8. 0 software, the maximum non lethal concentration (MN-LC) was simulated. Zebrafish raised in different concentrations of Wufang Babu Ointment solution for 24 h were placed under an anatomical microscope for taking photos, to analyze and calculate the incidence of skin damage in zebrafish. Based on the statistical analysis results of this indicator, the skin toxicity of Wufang Babu Ointment was evaluated. [Results] The MNLC of Wufang Babu Ointment on zebrafish was 671 µg/ mL; Wufang Babu Ointment can induce skin damage at the concentrations of 224 µg/mL (1/3 MNLC) and 671 µg/mL (MNLC). [Conclusions] Wufang Babu Ointment had certain skin toxicity to zebrafish. [ABSTRACT FROM AUTHOR]

The global circulation of clade 2.3.4.4b H5Ny highly pathogenic avian influenza viruses (HPAIVs) in poultry and wild birds, increasing mammal infections, continues to pose a public health threat and may even form a pandemic. An efficacious vaccine against H5Ny HPAIVs is crucial for emergency use and pandemic preparedness. In this study, we developed a parainfluenza virus 5 (PIV5)-based vaccine candidate expressing hemagglutinin (HA) protein of clade 2.3.4.4b H5 HPAIV, termed rPIV5-H5, and evaluated its safety and efficacy in mice and ferrets. Our results demonstrated that intranasal immunization with a single dose of rPIV5-H5 could stimulate H5-specific antibody responses, moreover, a prime-boost regimen using rPIV5-H5 stimulated robust humoral, cellular, and mucosal immune responses in mice. Challenge study showed that rPIV5-H5 prime-boost regimen provided sterile immunity against lethal clade 2.3.4.4b H5N1 virus infection in mice and ferrets. Notably, rPIV5-H5 prime-boost regimen provided protection in mice against challenge with lethal doses of heterologous clades 2.2, 2.3.2, and 2.3.4 H5N1, and clade 2.3.4.4h H5N6 viruses. These results revealed that rPIV5-H5 can elicit protective immunity against a diverse clade of highly pathogenic H5Ny virus infection in mammals, highlighting the potential of rPIV5-H5 as a pan-H5 influenza vaccine candidate for emergency use. [ABSTRACT FROM AUTHOR]

Influenza A viruses in animal reservoirs repeatedly cross species barriers to infect humans. Dogs are the closest companion animals to humans, but the role of dogs in the ecology of influenza viruses is unclear. H3N2 avian influenza viruses were transmitted to dogs around 2006 and have formed stable lineages. The long-term epidemic of avian-origin H3N2 virus in canines offers the best models to investigate the effect of dogs on the evolution of influenza viruses. Here, we carried out a systematic and comparative identification of the biological characteristics of H3N2 canine influenza viruses (CIVs) isolated worldwide over 10 years. We found that, during adaptation in dogs, H3N2 CIVs became able to recognize the human-like SAα2,6-Gal receptor, showed gradually increased hemagglutination (HA) acid stability and replication ability in human airway epithelial cells, and acquired a 100% transmission rate via respiratory droplets in a ferret model. We also found that human populations lack immunity to H3N2 CIVs, and even preexisting immunity derived from the present human seasonal influenza viruses cannot provide protection against H3N2 CIVs. Our results showed that canines may serve as intermediates for the adaptation of avian influenza viruses to humans. Continuous surveillance coordinated with risk assessment for CIVs is necessary. [ABSTRACT FROM AUTHOR]

Aging, subjected to scientific scrutiny, is extensively defined as a time-dependent decline in functions that involves the majority of organisms. The time-dependent accretion of cellular lesions is generally a universal trigger of aging, while mitochondrial dysfunction is a sign of aging. Dysfunctional mitochondria are identified and removed by mitophagy, a selective form of macroautophagy. Increased mitochondrial damage resulting from reduced biogenesis and clearance may promote the aging process. The primary purpose of this paper is to illustrate in detail the effects of mitophagy on aging and emphasize the associations between mitophagy and other signs of aging, including dietary restriction, telomere shortening, epigenetic alterations, and protein imbalance. The evidence regarding the effects of these elements on aging is still limited. And although the understanding of relationship between mitophagy and aging has been long-awaited, to analyze details of such a relationship remains the main challenge in aging studies. [ABSTRACT FROM AUTHOR]

To improve repeatability and efficiency and to simplify the operation procedure of capillary electrophoresis (CE), a pressurized CE system (p-CE) with injection valve sampling was developed. It consisted of one high-pressure pump, a six-port injection valve, a PEEK cross, a separation and back pressure capillary, an ultraviolet-visible detector and a high voltage power supply. The pressure-driven flow ranging from 4.5 nL min-1 to 0.81 µL min-1 in the separation capillary was produced by splitting to the flow from the high-pressure pumps (0.005-0.4 mL min-1). Nano-volume sample injection (<10 nL) was conducted by a micro-volume rotary injector (0.5-5 µL) with flow splitting. In the p-CE system, the new commercial capillary could be directly used without any wash, and the capillary-flush process between runs was also eliminated. In this case, the analytes were driven toward the outlet of the separation capillary by the pressurized flow, the electric field force and minute electroosmotic flow, and they were separated owing to the electrophoretic mobility. The p-CE system allows for the independent variation of the pressurized flow rate and electrical field and electrophoretic separation of good repeatability (below 3%) under high electrical fields (500-1000 V cm-1) and flow rate gradient modes. The feasibility of the p-CE system in real analysis was demonstrated by iodate quantification in iodized table salts. The separation of iodide and iodate was realized within 0.3 min, proving its high analytical speed. [ABSTRACT FROM AUTHOR]

[Objectives] To establish the quality standard of Fuyang Wenshen Granules and carry out the routine examination of granule type. [Methods] The particle size, water content, solubility, loading difference and microbial limit of Fuyang Wenshen Granules were examined, and TLC was used for qualitative identification of Astmgali Radix and Notoginseng Radix Et Rhizoma. [Results] The particle size, water content, solubility, loading difference, and microbial limit of Fuyang Wenshen Granules complied with the relevant provisions of the Pharmacopoeia of the People's Republic of China (2020). The TLC spots of Astmgali Radix and Notoginseng Radix Et Rhizoma were clearly colored, and negative had no interference. [Conclusions] The method was simple, accurate, reliable, specific and reproducible, and can effectively control the quality of Fuyang Wenshen Granules. [ABSTRACT FROM AUTHOR]

Salinity variation in ancient epicontinental seas has long been challenging to reconstruct. We determined salinity variation in the Late Pennsylvanian North American Midcontinent Sea based on normalized boron concentrations (B/Ga) and isotopic compositions (δ11B) of bulk shale. We analyzed the Stark Shale Member of the Missourian Stage Dennis Limestone cyclothem in the Charleston core (CC) from the Illinois Basin (eastward, more proximal) and the Iowa Riverton core (IRC) from the Midcontinent Shelf (westward, more distal). Both cores exhibited the same pattern of secular variation, with lower B/Ga and δ11B values at their base and top, recording less saline conditions during lower sea-level stands, and higher values in their middle part, recording more saline conditions at highstands--a pattern that conforms to conventional sequence stratigraphic and glacio-eustatic interpretations of Midcontinent cyclothems. At equivalent stratigraphic levels, B/Ga and δ11B values are systematically lower at CC relative to IRC, reflecting the generally lower salinity conditions of the Illinois Basin relative to the Midcontinent shelf, which is consistent with greater freshwater influence from the east due to heavy river runoff into the Appalachian foreland basin. Our findings serve to demonstrate the utility of paired B concentration and isotope data for paleosalinity analysis of deep-time shale formations. [ABSTRACT FROM AUTHOR]

The Cucurbitaceae includes important crops such as cucumber, melon, watermelon, squash and pumpkin. However, few genetic and genomic resources are available for plant improvement. Some cucurbit species such as cucumber have a narrow genetic base, which impedes construction of saturated molecular linkage maps. We report herein the development of highly polymorphic simple sequence repeat (SSR) markers originated from whole genome shotgun sequencing and the subsequent construction of a high-density genetic linkage map. This map includes 995 SSRs in seven linkage groups which spans in total 573 cM, and defines ∼680 recombination breakpoints with an average of 0.58 cM between two markers. These linkage groups were then assigned to seven corresponding chromosomes using fluorescent in situ hybridization (FISH). FISH assays also revealed a chromosomal inversion between Cucumis subspecies [C. sativus var. sativus L. and var. hardwickii (R.) Alef], which resulted in marker clustering on the genetic map. A quarter of the mapped markers showed relatively high polymorphism levels among 11 inbred lines of cucumber. Among the 995 markers, 49%, 26% and 22% were conserved in melon, watermelon and pumpkin, respectively. This map will facilitate whole genome sequencing, positional cloning, and molecular breeding in cucumber, and enable the integration of knowledge of gene and trait in cucurbits. [ABSTRACT FROM AUTHOR]

Avian influenza viruses (AIVs) are zoonotic viruses that exhibit a range of infectivity and severity in the human host. Severe human cases of AIV infection are often accompanied by neurological symptoms; however, the factors involved in the infection of the central nervous system (CNS) are not well known. In this study, we discovered that the avian-like sialic acid (SA)--a2,3-galactose ( a2,3-Gal) receptor is highly presented in mammalian (human and mouse) brains. In the generation of a mouse-adapted neurotropic H9N2 AIV (SD16-MA virus) in BALB/c mice, we identified key adaptive mutations in its hemagglutinin (HA) and polymerase basic protein 2 (PB2) genes that conferred viral replication ability in the mouse brain. The SD16-MA virus showed binding affinity for the avian-like SA--a2,3-Gal receptor, enhanced viral RNP polymerase activity, and increased viral protein production and transport that culminated in elevated progeny virus production and severe pathogenicity. We further established that host fragile X mental retardation protein (FMRP), a highly expressed protein in the brain that is physically associated with viral nucleocapsid protein (NP) to facilitate RNP assembly and export, was an essential host factor for the neuronal replication of neurotropic AIVs (H9N2, H5N1, and H10N7 viruses). Our study identified a mechanistic process for AIVs to acquire neurovirulence in mice. [ABSTRACT FROM AUTHOR]

Sepsis is a major cause of mortality in intensive care units, which results from a severely dysregulated inflammatory response that ultimately leads to organ failure. While antibiotics can help in the early stages, effective strategies to curtail inflammation remain limited. The high mobility group (HMG) proteins are chromosomal proteins with important roles in regulating gene transcription. While HMGB1 has been shown to play a role in sepsis, the role of other family members including HMGXB4 remains unknown. We found that expression of HMGXB4 is strongly induced in response to lipopolysaccharide (LPS)-elicited inflammation in murine peritoneal macrophages. Genetic deletion of Hmgxb4 protected against LPS-induced lung injury and lethality and cecal ligation and puncture (CLP)-induced lethality in mice, and attenuated LPSinduced proinflammatory gene expression in culturedmacrophages. By integrating genome-wide transcriptome profiling and a publicly available ChIP-seq dataset, we identified HMGXB4 as a transcriptional activator that regulates the expression of the proinflammatory gene, Nos2 (inducible nitric oxide synthase 2) by binding to its promoter region, leading to NOS2 induction and excessive NO production and tissue damage. Similar to Hmgxb4 ablation in mice, administration of a pharmacological inhibitor of NOS2 robustly decreased LPS-induced pulmonary vascular permeability and lethality in mice. Additionally, we identified the cell adhesion molecule, ICAM1, as a target of HMGXB4 in endothelial cells that facilitates inflammation by promoting monocyte attachment. In summary, our study reveals a critical role of HMGXB4 in exacerbating endotoxemia via transcriptional induction of Nos2 and Icam1 gene expression and thus targeting HMGXB4 may be an effective therapeutic strategy for the treatment of sepsis. [ABSTRACT FROM AUTHOR]

Avian influenza virus (AIV) can cross species barriers to infect humans and other mammals. However, these species-cross transmissions are most often dead-end infections due to host restriction. Current research about host restriction focuses mainly on the barriers of cell membrane, nuclear envelope, and host proteins; whether microRNAs (miRNAs) play a role in host restriction is largely unknown. In this study, we used porcine alveolar macrophage (PAM) cells as a model to elucidate the role of miRNAs in host range restriction. During AIV infection, 40 dysregulation expressed miRNAs were selected in PAM cells. Among them, two Sus scrofa (ssc; swine) miRNAs, ssc-miR-221-3p and ssc-miR-222, could inhibit the infection and replication of AIV in PAM cells by directly targeting viral genome and inducing cell apoptosis via inhibiting the expression of anti-apoptotic protein HMBOX1. Avian but not swine influenza virus caused upregulated expressions of ssc-miR-221-3p and ssc-miR- 222 in PAM cells. We further found that NF-B P65 was more effectively phosphorylated upon AIV infection and that P65 functioned as a transcription activator to regulate the AIV-induced expression of miR-221-3p/222. Importantly, we found that ssc-miR-221-3p and ssc-miR-222 could also be specifically upregulated upon AIV infection in newborn pig tracheal epithelial (NPTr) cells and also exerted anti-AIV function. In summary, our study indicated that miRNAs act as a host barrier during crossspecies infection of influenza A virus. [ABSTRACT FROM AUTHOR]

[Objectives] To study the long-term toxicity of Maxing Erchen Zhike granules to rats after intragastric administration, so as to provide reference for its preclinical safety evaluation. [Methods] Total 80 rats were randomly and evenly divided into high-dose group (1.2 mL/100 g, 120 g/kg), middle-dose group (96.0 g/kg), low-dose group (72.0 g/kg) and blank control group. The rats in the treatment groups were administered with corresponding doses of Maxing Erchen Zhike granules, and those in the blank control group were given with equal-amount normal saline. The administration lasted for 30 consecutive days. During the experiment, the rats' feed intake, activity, feces and other conditions and toxicity reactions were observed every day. After 24 h of the last administration, 12 rats (half male and half female) were randomly selected from each group. Each of the rats was anesthetized with 10% chloral hydrate solution (0.3 mL/100 g) through intraperitoneal injection and subjected to abdominal aorta blood collection (two tubes) for hematological examination and blood biochemical examination (serum). Then, the main organs of the rats were weighed, and pathological examinations were performed. After that, the man organs were weighed and pathological examination was performed. The remaining rats in each group were discontinued and observed for 14 d. On the 15th d, they were subjected to the same treatment, and the body weight, organ coefficients, hematological indices, blood biochemical indices and pathological indices were examined. [Results] After 30 d of administration, there was no abnormality in the appearance and behavior of the animals. There was no significant difference in the daily consumption of feed among the groups, and there was no special case of weight gain. Among the blood biochemical indices, the ALB and ALT levels of each administration group were significantly different from those of the blank control group (P < 0.05). The results of histopathological examination show that there was one case of interstitial pneumonia in each of the high-dose group, middle-dose group and blank control group. After 14 d that the administration was stopped, one case of focal myocarditis appeared in the high-dose group, and one case of interstitial pneumonia appeared in the middle-dose group. [Conclusions] Maxing Erchen Zhike granules are safe to be administered to rats at 100 times the clinical dose, and there should be no safety hazards clinically when used at conventional doses. [ABSTRACT FROM AUTHOR]

Equine-origin H3N8 and avian-origin H3N2 canine influenza viruses (CIVs) prevalent in dogs are thought to pose a public health threat arising from intimate contact between dogs and humans. However, our understanding of CIV virulence is still limited. Influenza A virus PA-X is a fusion protein encoded in part by a +1 frameshifted open reading frame (X-ORF) in segment 3. The X-ORF can be translated in full-length (61 amino acids) or truncated (41 amino acids) form. Genetic analysis indicated that the X-ORFs of equine H3N8 and avian H3N2 influenza viruses encoded 61 amino acids but were truncated after introduction into dogs. To determine the effect of PA-X truncation on the biological characteristics of CIVs, we constructed four recombinant viruses on H3N8 and H3N2 CIV backgrounds bearing truncated or full-length PA-Xs. We observed that truncation of PA-X increased growth of both H3N8 and H3N2 CIVs in MDCK cells and suppressed expression from co-transfected plasmids in MDCK cells. Furthermore, truncation of PA-X enhanced viral pathogenicity in dogs as shown by aggravated clinical symptoms and histopathological changes, increased viral replication in the respiratory system, and prolonged virus shedding. Additionally, CIVs with truncated PA-Xs were transmitted more efficiently in dogs. Global gene expression profiling of the lungs of infected dogs revealed that differentially expressed genes were mainly associated with inflammatory responses, which might contribute to the pathogenicity of PA-X-truncated CIVs. Our findings revealed that truncation of PA-X might be important for the adaptation of influenza viruses to dogs. [ABSTRACT FROM AUTHOR]

In the 21st century, the emergence of H7N9 and H1N1/2009 influenza viruses, originating from animals and causing severe human infections, has prompted investigations into the genetic alterations required for cross-species transmission. We previously found that replacement of the human-origin PA gene segment in avian influenza virus (AIV) could overcome barriers to cross-species transmission. Recently, it was reported that the PA gene segment encodes both the PA protein and a second protein, PA-X. Here, we investigated the role of PA-X. We found that an H9N2 avian influenza reassortant virus bearing a human-origin H1N1/2009 PA gene was attenuated in mice after the loss of PA-X. Reverse genetics analyses of PA-X substitutions conserved in human influenza viruses indicated that R195K, K206R, and P210L substitutions conferred significantly increased replication and pathogenicity on H9N2 virus in mice and ferrets. PA-X R195K was present in all human H7N9 and H1N1/2009 viruses and predominated in human H5N6 viruses. Compared with PA-X 195R, H7N9 influenza viruses bearing PA-X 195K showed increased replication and transmission in ferrets. We further showed that PA-X 195K enhanced lung inflammatory responses, potentially due to decreased host shutoff function. A competitive transmission study in ferrets indicated that 195K provides a replicative advantage over 195R in H1N1/2009 viruses. In contrast, PA-X 195K did not influence the virulence of H9N2 AIV in chickens, suggesting that the effects of the substitution were mammal specific. Therefore, future surveillance efforts should scrutinize this region of PA-X because of its potential impact on cross-species transmission of influenza viruses. [ABSTRACT FROM AUTHOR]

This work investigated the feasibility of manufacturing composite crosslaminated timber (CCLT) from bamboo and a western hemlock and amabilis fir mix (hem-fir). Bamboo parallel strand lumber (bamboo PSL) was first fabricated as the surface laminae of CCLT. No. 2 and Btr 2×6 western hem-fir dimension lumber was imported from British Columbia (BC), Canada. The lumber was then sorted and further dried to form the inner laminae of CCLT. Multiple 800 × 800-mm CLT billets were fabricated using a commercial single-component polyurethane (PUR) adhesive. Block shear and delamination tests were conducted to examine the bond quality and durability of bamboo-wood CCLT. The results demonstrated that it is technically feasible to manufacture bamboo-wood CCLT with acceptable bond quality (shear strength, wood failure percentage, etc.) and delamination. Further studies should focus on the optimization of process parameters for manufacturing and surface treatment to improve the bond quality and durability and quantify the mechanical performance of bamboo-wood CCLT with a full-size production trial. [ABSTRACT FROM AUTHOR]

During 2012-2017, we collected throat swabs from dogs in China to characterize canine influenza virus (CIV) A(H3N2) isolates. A new antigenically and genetically distinct CIV H3N2 clade possessing mutations associated with mammalian adaptation emerged in 2016 and replaced previously circulating strains. This clade probably poses a risk for zoonotic infection. [ABSTRACT FROM AUTHOR]

In response to vascular injury, vascular smooth muscle cells (VSMCs) may switch from a contractile to a proliferative phenotype thereby contributing to neointima formation. Previous studies showed that the long noncoding RNA (lncRNA) NEAT1 is critical for paraspeckle formation and tumorigenesis by promoting cell proliferation and migration. However, the role of NEAT1 in VSMC phenotypic modulation is unknown. Herein we showed that NEAT1 expression was induced in VSMCs during phenotypic switching in vivo and in vitro. Silencing NEAT1 in VSMCs resulted in enhanced expression of SM-specific genes while attenuating VSMC proliferation and migration. Conversely, overexpression of NEAT1 in VSMCs had opposite effects. These in vitro findings were further supported by in vivo studies in which NEAT1 knockout mice exhibited significantly decreased neointima formation following vascular injury, due to attenuated VSMC proliferation. Mechanistic studies demonstrated that NEAT1 sequesters the key chromatin modifier WDR5 (WD Repeat Domain 5) from SM-specific gene loci, thereby initiating an epigenetic "off" state, resulting in down-regulation of SM-specific gene expression. Taken together, we demonstrated an unexpected role of the lncRNA NEAT1 in regulating phenotypic switching by repressing SM-contractile gene expression through an epigenetic regulatory mechanism. Our data suggest that NEAT1 is a therapeutic target for treating occlusive vascular diseases. [ABSTRACT FROM AUTHOR]

Segment reassortment and base mutagenesis of influenza A viruses are the primary routes to the rapid evolution of high-fitness virus genotypes. We recently described a predominant G57 genotype of avian H9N2 viruses that caused countrywide outbreaks in chickens in China during 2010 to 2013, which led to the zoonotic emergence of H7N9 viruses. One of the key features of the G57 genotype is the replacement of the earlier A/chicken/Beijing/1/1994 (BJ/94)-like M gene with the A/quail/Hong Kong/G1/1997 (G1)-like M gene of quail origin. We report here the functional significance of the G1-like M gene in H9N2 viruses in conferring increased infection severity and infectivity in primary chicken embryonic fibroblasts and chickens. H9N2 virus housing the G1-like M gene, in place of the BJ/94-like M gene, showed an early surge in viral mRNA and viral RNA (vRNA) transcription that was associated with enhanced viral protein production and with an early elevated release of progeny virus comprising largely spherical rather than filamentous virions. Importantly, H9N2 virus with the G1-like M gene conferred extrapulmonary virus spread in chickens. Five highly represented signature amino acid residues (37A, 95K, 224N, and 242N in the M1 protein and 21G in the M2 protein) encoded by the prevalent G1-like M gene were demonstrated to be prime contributors to enhanced infectivity. Therefore, the genetic evolution of the M gene in H9N2 virus increases reproductive virus fitness, indicating its contribution to the rising virus prevalence in chickens in China. [ABSTRACT FROM AUTHOR]

Ginsenoside Rg3, a bioactive constituent isolated from Panax ginseng, exhibits antitumorigenic, antioxidative, antiangiogenic, neuroprotective and other biological activities are associated with the regulation of multiple genes. DNA methylation patterns, particularly those in the promoter region, affect gene expression, and DNA methylation is catalyzed by DNA methylases. However, whether ginsenoside Rg3 affects DNA methylation is unknown. High performance liquid chromatography assay, MspI/HpaII polymerase chain reaction (PCR) and reverse transcription-quantitative PCR were performed to assess DNA methylation. It was demonstrated that 20(S)-ginsenoside Rg3 treatment resulted in increased inhibition of cell growth, compared with treatment with 20(R)-ginsenoside Rg3 in the human HepG2 hepatocarcinoma cell line. It was additionally revealed that treatment with 20(S)-ginsenoside Rg3 reduced global genomic DNA methylation, altered cystosine methylation of the promoter regions of P53, B cell lymphoma 2 and vascular endothelial growth factor, and downregulated the expression of DNA methyltransferase (DNMT) 3a and DNMT3b more than treatment with 20(R)-ginsenoside Rg3 in HepG2 cells. These results revealed that the modulation of DNA methylation may be important in the pharmaceutical activities of ginsenoside Rg3. [ABSTRACT FROM AUTHOR]

Since May 2014, highly pathogenic avian influenza H5N6 virus has been reported to cause six severe human infections three of which were fatal. The biological properties of this subtype, in particular its relative pathogenicity and transmissibility in mammals, are not known. We characterized the virus receptor-binding affinity, pathogenicity, and transmissibility in mice and ferrets of four H5N6 isolates derived from waterfowl in China from 2013-2014. All four H5N6 viruses have acquired a binding affinity for human-like SAα2,6Gal-linked receptor to be able to attach to human tracheal epithelial and alveolar cells. The emergent H5N6 viruses, which share high sequence similarity with the human isolate A/Guangzhou/39715/2014 (H5N6), were fully infective and highly transmissible by direct contact in ferrets but showed less-severe pathogenicity than the parental H5N1 virus. The present results highlight the threat of emergent H5N6 viruses to poultry and human health and the need to closely track their continual adaptation in humans. [ABSTRACT FROM AUTHOR]

Background: In epidemic regions of the world, brucellosis is a reemerging zoonosis with minimal mortality but is a serious public hygiene problem. Currently, there are various methods for brucellosis diagnosis, however few of them are available to be used to diagnose, especially for serious cross-reaction with other bacteria.Method: To overcome this disadvantage, we explored a novel multi-epitope recombinant protein as human brucellosis diagnostic antigen. We established an indirect enzyme-linked immunosorbent assay (ELISA) based on this recombinant protein. 248 sera obtained from three different groups including patients with brucellosis (146 samples), non-brucellosis patients (82 samples), and healthy individuals (20 samples) were tested by indirect ELISA. To evaluate the assay, a receiver-operating characteristic (ROC) analysis and immunoblotting were carried out using these characterized serum samples.Results: For this test, the area under the ROC curve was 0.9409 (95 % confidence interval, 0.9108 to 0.9709), and a sensitivity of 88.89 % and a specificity of 85.54 % was given with a cutoff value of 0.3865 from this ROC analysis. The Western blot results indicate that it is feasible to differentiate human brucellosis and non-brucellosis with the newly established method based on this recombinant protein.Conclusion: Our results obtained high diagnostic accuracy of the ELISA assay which encourage the use of this novel recombinant protein as diagnostic antigen to implement serological diagnosis of brucellosis. [ABSTRACT FROM AUTHOR]

Monitoring Volatile organic compounds (VOCs) was a very important measure for preventing environmental pollution, therefore, a mini gas chromatography (GC) flame ionization detector (FID) system integrated with a mini H2 generator and a micro GC column was developed for environmental VOC monitoring. In addition, the mini H2 generator was able to make the system explode from far away due to the abandoned use of a high pressure H2 source. The experimental result indicates that the fabricated mini GC FID system demonstrated high repeatability and very good linear response, and was able to rapidly monitor complicated environmental VOC samples. [ABSTRACT FROM AUTHOR]

Background: The close contact between cats and humans poses a threat to public health because of the potential zoonotic transmission of influenza viruses to humans. Therefore, we examined the seroprevalence of pandemic H1N1/09, canine H3N2, and human H3N2 viruses in pet cats in northern China from 2010 to 2014. Finding: Of 1794 serum samples, the seropositivity rates for H1N1/09, canine H3N2, and human H3N2 were 5.7%, 0.7%, and 0.4%, respectively. The seropositivity rate for H1N1/09 in cats was highest in 2010 (8.3%), and then declined continuously thereafter. Cats older than 10 years were most commonly seropositive for the H1N1/09 virus. Conclusions: Our findings emphasize the need for continuous surveillance of influenza viruses in cats in China. [ABSTRACT FROM AUTHOR]

Genetic and phylogenetic analyses suggest that the pandemic H1N1/2009 virus was derived from well-established swine influenza lineages; however, there is no convincing evidence that the pandemic virus was generated from a direct precursor in pigs. Furthermore, the evolutionary dynamics of influenza virus in pigs have not been well documented. Here, we subjected a recombinant virus (rH1N1) with the same constellation makeup as the pandemic H1N1/2009 virus to nine serial passages in pigs. The severity of infection sequentially increased with each passage. Deep sequencing of viral quasispecies from the ninth passage found five consensus amino acid mutations: PB1 A469T, PA 1129T, NA N329D, NS1 N205K, and NEP T48N. Mutations in the hemagglutinin (HA) protein, however, differed greatly between the upper and lower respiratory tracts. Three representative viral clones with the five consensus mutations were selected for functional evaluation. Relative to the parental virus, the three viral clones showed enhanced replication and polymerase activity in vitro and enhanced replication, pathogenicity, and transmissibility in pigs, guinea pigs, and ferrets in vivo. Specifically, two mutants of rH1N1 (PB1 A469T and a combination of NS1 N205K and NEP T48N) were identified as determinants of transmissibility in guinea pigs. Crucially, one mutant viral clone with the five consensus mutations, which also carried D187E, K211E, and S289N mutations in its HA, additionally was able to infect ferrets by airborne transmission as effectively as the pandemic virus. Our findings demonstrate that influenza virus can acquire viral characteristics that are similar to those of the pandemic virus after limited serial passages in pigs. [ABSTRACT FROM AUTHOR]

The D222N hemagglutinin (HA) mutation within the receptor-binding site was detected with higher frequencies in severe cases of 2009 pandemic H1N1 (pdmH1N1) infections. The impact of this mutation was investigated in vitro and in vivo using recombinant viruses. The recombinant D222N virus grew to significantly lower viral titers than the WT in A549 but not in MDCK cells. A dose-dependent glycan array analysis with the D222N virus showed a modest increase in the binding avidity to human-like (α-2,6 sialylated glycan) receptors and avian-like (α-2,3 sialylated glycan) receptors than the WT virus. The D222N HA mutation resulted in slight weight loss, lower lung titers, inflammatory cytokines and alveolar inflammation in mice than the WT virus. This may or may not be associated with severe clinical outcomes reported in humans. [ABSTRACT FROM AUTHOR]

The article discusses the study of the impact of naturally occurring mutations in the PA gene in enhancing the virulence of Pandemic H1N1/09 Influenza Virus in mice. Topics discussed include examination of molecular basis of virulence of the viruses by reverse genetics on the basis of two H1N1/09 virus isolates including A/California/04/2009 and A/swine/Shandong/731/2009, effect of the introduction of virulence markers on pathogenicity and evaluation of virulence determinants of H1N1/09.

Highly pathogenic avian influenza H5N1 virus clades 2.3.4, 2.3.2, and 7 are the dominant cocirculating H5N1 viruses in poultry in China. However, humans appear to be clinically susceptible mostly to the 2.3.4 virus clade. Here, we demonstrated that A549 cells and human macrophages infected with clade 2.3.4 viruses produced significantly more viruses than those infected with the other two clades. Likewise, clade 2.3.4-infected macrophages caused the most severe cellular damage and strongest proinflammatory response. [ABSTRACT FROM AUTHOR]

An avian-origin human-infecting influenza (H7N9) virus was recently identified in China. We have evaluated the viral hemagglutinin (HA) receptor-binding properties of two human H7N9 isolates, A/Shanghai/1/2013 (SH-H7N9) (containing the avian-signature residue Gln226) and A/Anhui/1/2013 (AH-H7N9) (containing the mammalian-signature residue Leu226). We found that SH-H7N9 HA preferentially binds the avian receptor analog, whereas AH-H7N9 HA binds both avian and human receptor analogs. Furthermore, an AH-H7N9 mutant HA (Leu226 → Gln) was found to exhibit dual receptor-binding property, indicating that other amino acid substitutions contribute to the receptor-binding switch. The structures of SH-H7N9 HA, AH-H7N9 HA, and its mutant in complex with either avian or human receptor analogs show how AH-H7N9 can bind human receptors while still retaining the avian receptor-binding property. [ABSTRACT FROM AUTHOR]

H9N2 influenza viruses with an A316S substitution in hemagglutinin (HA) and a shorter neuraminidase (NA) stalk have become predominant in China. The A316S was shown to increase HA cleavage efficiency when combined with short stalk NA, and the short stalk NA improved NA enzyme activity and release of virus from erythrocytes. Single mutations or combinations of these mutations strengthened the virulence of H9N2 virus in chickens and mice. [ABSTRACT FROM AUTHOR]

To determine risk for avian influenza virus infection, we conducted serologic surveillance for H5 and H9 subtypes among poultry workers in Beijing, China, 2009-2010, and assessed workers' understanding of avian influenza. We found that poultry workers had considerable risk for infection with H9 subtypes. Increasing their knowledge could prevent future infections. [ABSTRACT FROM AUTHOR]

H9N2 influenza viruses have been circulating worldwide in multiple avian species and have repeatedly infected humans to cause typical disease. The continued avian-to-human interspecies transmission of H9N2 viruses raises concerns about the possibility of viral adaption with increased virulence for humans. To investigate the genetic basis of H9N2 influenza virus host range and pathogenicity in mammals, we generated a mouse-adapted H9N2 virus (SD16-MA) that possessed significantly higher virulence than wide-type virus (SD16). Increased virulence was detectable after 8 sequential lung passages in mice. Five amino acid substitutions were found in the genome of SD16-MA compared with SD16 virus: PB2 (M147L, V250G and E627K), HA (L226Q) and M1 (R210K). Assessments of replication in mice showed that all of the SD16-MA PB2, HA and M1 genome segments increased virus replication; however, only the mouse-adapted PB2 significantly increased virulence. Although the PB2 E627K amino acid substitution enhanced viral polymerase activity and replication, none of the single mutations of mouse adapted PB2 could confer increased virulence on the SD16 backbone. The combination of M147L and E627K significantly enhanced viral replication ability and virulence in mice. Thus, our results show that the combination of PB2 amino acids at position 147 and 627 is critical for the increased pathogenicity of H9N2 influenza virus in mammalian host. [ABSTRACT FROM AUTHOR]