Gut microbial changes in a specialist blister beetle larvae and their nutritional metabolic characteristics.

Insect gut microbiota and their metabolites play a significant role in the shaping of hosts' diets and feeding habits. We conducted 16S rDNA amplicon sequencing on the gut microbiota of specialist blister beetle larvae that feed on locust eggs and artificial food at different instars, to explore the relationship between gut microbiota and the specialized feeding habit of the blister beetle larvae. There is no significant difference in the gut microbial structure among the second to the fourth instar larvae under the same rearing conditions, but the gut microbial structure of the first instar larvae was significantly different from the second to the fourth instar larvae fed by different diets. Bacteria associated with polysaccharide utilization are relatively barren in first instar larvae. Compared to the carbohydrate content between the artificial diet and locust eggs, we speculate that an excessive amount of polysaccharides in the artificial diet may be detrimental to the growth and development of first instar larvae. Gut microbiota of the second to the fourth instar larvae fed with different diets significantly differ in microbial community structure. The different bacteria, especially the metabolism-related intestinal bacteria in locust eggs-fed larvae, may help the hosts adapt to the environment and contribute to the production of active ingredients. The relative abundance of polysaccharide utilization-related bacteria was significantly higher in the artificial diet-fed larvae compared to the locust eggs-fed larvae, which showed the same result when compared to the first instar larvae. Changes in gut microbes of blister beetle larvae and their metabolic inferences could enrich our understanding of the nutritional requirements of the specialist and help optimize the artificial diet of medicinal cantharides.

Exosomal miR-150 partially attenuated acute lung injury by mediating microvascular endothelial cells and MAPK pathway.

BACKGROUND: Acute lung injury (ALI) is a respiratory disease with high morbidity and mortality rates. Currently, there is no effective treatment to complement mechanical ventilation. Exosomes and microRNAs (miRNAs) are promising agents for the management of this disease. METHODS: Exosomes were isolated from mouse bone marrow stromal stem cells (BMSCs). The levels of two miRNAs, miR-542-3P and miR-150, in exosomes were determined using RT-PCR, and miR-150 was selected for further study. ALI model was established in mice using lipopolysaccharides, and then, they were treated with saline, exosomes, miRNA agomirs, or miRNA antagomirs. The concentrations of TNF-α, IL-6, and IL-1ß and the number of neutrophils and macrophages in the bronchoalveolar lavage fluid were measured. The wet/dry weight ratio of the lung tissue was calculated, and tissue pathology and apoptosis were observed using hematoxylin and eosin and terminal deoxynucleotidyl transferase dUTP nick-end labeling staining. CD34 and VE-cadherin expression was detected using immunofluorescence. Proteins associated with apoptosis and MAPK signaling were detected using Western blotting, and miR-150 expression in lung tissue was evaluated using RT-PCR. RESULTS: We successfully isolated BMSCs and exosomes and showed that the level of miR-150 was significantly higher than that of miR-542-3p. Exosomes and miR-150 reduced inflammation and lung edema while maintaining the integrity of the alveolar structure. They also mitigated microvascular endothelial cell injury by regulating the caspase-3, Bax/Bcl-2, and MAPK signaling. CONCLUSIONS: Exosomal miR-150 attenuates lipopolysaccharide-induced ALI through the MAPK pathway.

De novo transcriptome assemblies of Epicauta tibialis provide insights into the sexual dimorphism in the production of cantharidin.

Blister beetles have medicinal uses for their defensive secretion cantharidin, which has curative effects on many cancers and other diseases. It was demonstrated that sexual dimorphism exists in the production of cantharidin between male and female adults. This study performed a de novo assembly of Epicauta tibialis transcriptomes and analyzed the differentially expressed genes (DEGs) between male and female adults to help to find genes and pathways associated with cantharidin biosynthesis. A total of 99,295,624 paired reads were generated, and more than 7 Gb transcriptome data for each sample were obtained after trimming. The clean data were used to de novo assemble and then cluster into 27,355 unigenes, with a mean length of 1442 bp and an N50 of 2725 bp. Of these, 14,314 (52.33%) unigenes were annotated by protein databases. Differential expression analysis identified 284 differentially expressed genes (DEGs) between male and female adults. Nearly 239 DEGs were up-regulated in male adults than in female adults, while 45 DEGs were down-regulated. The Kyoto Encyclopedia of Gene and Genomes pathway enrichment manifested that seven up-regulated DEGs in male adults were assigned to the terpenoid biosynthesis pathway, to which 19 unigenes were annotated. The DEGs in the terpenoid biosynthesis pathway between male and female adults may be responsible for the sexual dimorphism in cantharidin production. The up-regulated genes assigned to the pathway in male adults may play a significant role in cantharidin biosynthesis, and its biosynthesis process is probably via the mevalonate pathway. The results would be helpful to better understand and reveal the complicated mechanism of the cantharidin biosynthesis.

The complete mitochondrial genome of Asian short-toed Lark Alaudala cheleensis (Aves: Passeriformes: Alaudidae).

We sequenced the mitochondrial genome of Asian short-toed Lark Alaudala cheleensis using the next-generation sequencing. The circular genome is 16,914 bp long, encoding 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), 2 ribosomal RNAs (rRNAs), and there are two control regions, which is similar to the common type suggested as ancestral for birds but has a 1126 bp control region and a 236-bp remnant control region. The phylogenetic analysis of published lark mitogenomes reveals the top phylogenetic position of A. cheleensis in Alaudidae.

The complete mitochondrial genome of Daurian redstart Phoenicurus auroreus (Aves: Passeriformes: Muscicapidae).

We sequenced the mitochondrial genome of Daurian redstart Phoenicurus auroreus using the next-generation sequencing. The circular genome is 16,832 bp long, encoding 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), 2 ribosomal RNAs (rRNAs), and a control region. The lengths of rrnL and rrnS were determined to be 1,597 and 984 bp, respectively. The phylogenetic analysis of 18 mitogenomes of Muscicapidae supports monophylies of all genera, including P. auroreus. The information would be useful for understanding the phylogeny and evolution of Muscicapidae.

The complete mitochondrial genome of the Eurasian wryneck Jynx torquilla (Aves: Piciformes: Picidae) and its phylogenetic inference.

The Eurasian Wryneck is a species of wryneck woodpecker breeding in temperate regions of Europe and Asia. We sequenced the mitochondrial genome of Jynx torquilla (Aves, Piciformes, Picidae) using the next generation sequencing. The circular genome is 16,832 bp long, encoding 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), two ribosomal RNAs (rRNAs), and two control regions. Gene order and orientation are similar to the most common type suggested as ancestral for birds but have a 1,221 bp control region and a 60 bp remnant control region. Phylogenetic analyses of 17 piciform taxa, based on both nucleotide and amino acid sequences of mitochondrial PCGs, strongly support the monophyly of Picidae. All phylogenetic trees indicate that the subfamily Jynginae is a monophyletic lineage sister to other woodpeckers, including monophyletic Picinae. Only the Bayes inferred tree based on the nucleotide dataset, recovered Picumninae as monophyletic. These findings will be helpful for the understanding of the phylogeny and evolution of Picidae.

Accumulated Clinical Experiences from Successful Treatment of 1377 Severe and Critically Ill COVID-19 Cases.

In late December 2019, COVID-19 was firstly recognized in Wuhan, China and spread rapidly to all of the provinces of China. The West Campus of Wuhan Union Hospital, the designated hospital to admit and treat the severe and critically ill COVID-19 cases, has treated a large number of such patients with great success and obtained lots of valuable experiences based on the Chinese guideline (V7.0). To standardize and share the treatment procedures of severe and critically ill cases, Wuhan Union Hospital has established a working group and formulated an operational recommendation, including the monitoring, early warning indicators, and several treatment principles for severe and critically ill cases. The treatment experiences may provide some constructive suggestions for treating the severe and critically ill COVID-19 cases all over the world.

Sampling and analysis of airborne ammonia in workplaces of China.

OBJECTIVES: With the increasing demand for the detection of occupational hazard factors in workplaces, the national standard determination method for ammonia (sampling with absorbing solution-analysis with Nessler reagent spectrophotometry) in the air of workplace presents many drawbacks during application in China. This review summarized the improvement and the alternate methods of the current sampling and analysis procedures for ammonia, aiming to provide reference to establish an appropriate method for the determination of ammonia in workplace air. METHODS: Scientific publications in English and Chinese and the standard methods of the Deutsche Forschungsgemeinschaft (DFG) in Germany, the National Institute for Occupational Safety and Health (NIOSH) and Occupational Safety and Health Administration (OSHA) in the United States, and Ministry of Health in China for airborne ammonia collection and analysis in the workplace were reviewed. RESULTS: The measures to improve the current sampling and analysis procedures for ammonia in China were firstly summarized. For sampling, the decrease of absorbing solution concentration and the methanesulfonic acid solution as the alternate sampling solution were suggested. For analysis, the anti-interference measures and the optimum reaction condition between ammonia and Nessler reagent were discussed. The alternate methods including sampling conducted using solid sorbent tubes and analysis performed by ion chromatography were then considered for the determination of ammonia. CONCLUSIONS: The methods-sampling with acid-treated solid sorbent tubes and analysis with ion chromatography-were more suitable for the determination of ammonia in workplace air. However, some details about ammonia sampling and analysis still need further investigation.

Activation of Alpha-7 Nicotinic Acetylcholine Receptors (α7nAchR) Promotes the Protective Autophagy in LPS-Induced Acute Lung Injury (ALI) In Vitro and In Vivo.

The release of inflammatory cytokines and chemokines and autophagy has been reported to be involved in the pathogenic mechanism of acute lung injury (ALI). Reportedly, alpha-7 nicotinic acetylcholine receptors (α7nAchR) might play a protective role in LPS-induced ALI. In the current research, we established LPS-induced ALI model in mice and α7nAchR agonist PNU-282987 improved LPS-induced injury. In MH-S cells, LPS stimulation inhibited, whereas α7nAchR agonist PNU-282987 enhanced the autophagy. α7nAchR agonist PNU-282987 protected MH-S cells from LPS-induced inflammation by reducing the concentrations of IL-6, TNF-α, and IL-1ß. Finally, LPS stimulation dramatically inhibited MH-S cell viability but enhanced cell apoptosis, whereas PNU-282987 treatment exerted opposite effects; α7nAchR might regulate the cellular homeostasis via affecting the crosstalk between the autophagy and apoptosis in MH-S cells; in other words, α7nAChR agonist enhances MH-S cell autophagy and inhibits MH-S cell apoptosis. In conclusion, α7nAchR promote the protective autophagy in LPS-induced ALI model in mice and MH-S cells. The application of α7nAchR agonist is considered a potent target for LPS-induced ALI, which needs further clinical investigation.

Electricity generation, salinity, COD removal and anodic biofilm microbial community vary with different anode CODs in a microbial desalination cell for high-salinity mustard tuber wastewater treatment.

A three-chamber microbial desalination cell (MDC) was constructed for high-salinity mustard tuber wastewater (MTWW) treatment. The effect of anode COD on electricity generation, salinity, COD removal and the anodic biofilm microbial community in MDC for the MTWW treatment was investigated. The results showed that electricity generation was better when the anode COD was 900 mg L-1 versus when it was 400 or 1400 mg L-1. The ionic strength and conductivity of the anolyte were higher than those at 400 mg L-1; thus, the ohmic internal resistance was lower. In addition, the mass transfer internal resistance was lower than that at 1400 mg L-1, which made the system internal resistance the lowest; consequently, the voltage and power density were the highest. The output voltage, power density and coulombic efficiency of the 1000 Ω external resistors were 555 mV, 3.03 W m-3 and 26.5% ± 0.4%, respectively. Desalination was the highest when the anode COD was 400 mg L-1. The lowest ionic strength and osmotic pressure of the anolyte resulted in the strongest osmosis, thereby producing the highest desalination rate; the desalination rate was 5.33 mg h-1. When MDC was coupled with the dual-chamber microbial fuel cell (MFC), the desalinated MTWW could be used as the anode substrate of the MFC; its high COD could be removed continuously, and the COD removal values were 86.2% ± 2.5%, 83.0% ± 2.0% and 84.3% ± 2.4%. High-throughput sequencing analysis indicated that hydrolytic and fermentative bacteria were the core anode bacteria of MDC. The abundances of electrochemically active bacteria in the anode biofilms of the three groups were 11.78% (400 mg L-1 COD), 14.06% (900 mg L-1 COD) and 13.68% (1400 mg L-1 COD). Therefore, the differences in anode CODs impacted the abundance of electrochemically active bacteria, which led to differences in electricity generation performances.

Rapid Detection of Atrazine at Workplace with Near-Infrared Spectroscopy.

As a wildly used herbicide, Atrazine is mainly produced in China. In order to strengthen the routine detection of Atrazine exposure concentration and protect the health of occupational contact workers, it's of great importance to develop on-site rapid detection method. A self-assembled near infrared spectrometer was used to record spectra of laboratory prepared atrazine solutions with concentration range from 10 to 1 000 mg·L-1. The influences of different pretreatment methods, such as multiplicative scatter correction, standard normal variate, first order derivative (D1), second order derivative and their combinations, different variable selection methods, such as competitive adaptive reweighted sampling (CARS) and genetic algorithm (GA), different regression methods, such as partial least square (PLS) and support vector regression(nu-SVR), on the model prediction accuracy were investigated. Results show that D1 is the best pretreatment method; GA obtain better results than CARS on selecting highly related spectral variables; nu-SVR model perform better than PLS model. The nu-SVR model constructed with 16 spectral variables selected by GA obtained the best results, whose coefficient of determination for calibration, the coefficient of determination for validation, root mean square error of calibration, root mean square error of validation (RMSEV) and residual validation deviation (defined as SD/RMSEV where SD denotes standard deviation) are 1, 0.99, 17.54 mg·L-1, 25.42 mg·L-1 and 11.43, respectively. These results indicate near infrared spectroscopy combined with chemometrics has great potential to quantify Atrazine concentration at workplace. This research explores the feasibility of quantification Atrazine at workplace with near infrared spectroscopy for the first time, which has great reference value for similar work in the future.

[An analysis of characteristics of exposure to nanoparticles in a workplace manufacturing iron oxide nanoparticles].

OBJECTIVE: To investigate the characteristics of exposure to iron oxide nanoparticles in workplace. METHODS: The real-time particle number (NC), surface area (SAC), and mass (MC) concentrations of nanoparticles were measured in various locations of a selected workplace manufacturing iron oxide nanoparticles. The collected particles were analyzed for morphology and elemental composition. RESULTS: The average NCs and SACs in milling site (16,566 pt/cm3, 106.082 µm2/cm3), packaging site (12,386 pt/cm3, 89.861 µm2/cm3), shipping site (13,808 pt/cm3, 102.071 µm2/cm3), and product storage room (17,192 pt/cm, 115.044 µm2/cm3) of the yellow powder (α-Fe2O3 . nH2O) were all significantly higher than the workplace background concentrations (11,420 pt/cm3, 85.026 µm2/cm3) (all P<0.05). The NC was highly correlated with the SAC (r= 0.784), while both NC and SAC were loosely correlated with the MC (r1=0.323, r2=0.331). Scanning electron microscopy revealed a spindle-like shape of the iron oxide nanoparticle; the chemical composition of the collected particles contained 19.33 weight percent iron (Fe). CONCLUSION: The milling site and product storage room of the yellow powder are exposed to a higher concentration of nanoparticles, which are mainly composed of iron oxide nanoparticles. The NC is highly correlated with the SAC.

[Determination of 2, 4-dichlorophenoxyacetic acid in air of workplace by high-performance liquid chromatography].

OBJECTIVE: To develop a method for determination of 2, 4-dichlorophenoxyacetic acid (2, 4-D) in the air of workplace by high-performance liquid chromatography. METHODS: 2, 4-D was collected by ultrafine glass filters, desorbed by methanol, separated by a C18 column, and detected by a UV detector. Identification and quantification of 2, 4-D were performed by retention time and peak areas, respectively. RESULTS: The linear range of the test was 2∼200 µg/ml; the elution efficiency was 94.6%- 95.9%; the limit of detection (S/N = 3) was 0.034 µg/ml (injection volume of 20 µl eluant); the lower limit of quantification (S/N = 10) was 0.11 µg/ml; the minimum detectable concentration was 0.011 mg/m(3); the minimum quantifiable concentration was 0.037 mg/m(3) (with sampled air volume of 45 L). CONCLUSION: This method is convenient and simple in sample collection and preparation, and satisfies all methodological requirements. Therefore, this method is useful for the determination of 2, 4-D in the air of workplace.

[Method for determining brodifacoum in workplace air by high-performance liquid chromatography].

OBJECTIVE: To establish a method for determining brodifacoum in workplace air by high-performance liquid chromatography (HPLC). METHODS: Brodifacoum in workplace air was collected with a polytetrafluoroethylene filter and desorbed by mixed solution of methanol and dichloromethane (20:80, V:V), and was then separated using an ODS column and determined by an ultraviolet detector; retention time was used for identification, and peak area was used for quantification. RESULTS: The concentration of brodifacoum showed a linear relationship with peak area within 0.2∼10.0 µg/ml; the elution efficiency was 91.6%∼95.1%; the detection limit was 0.08 µg/ml (injection volume: 20 µl eluate); the minimum detectable concentration was 0.000 67 mg/m(3) (calculated by 240 L air sample). CONCLUSION: This HPLC method is convenient and simple for air collection and sample preparation and meets the methodological requirements. Therefore, this method can be used for the determination of brodifacoum in workplace air.

Terpene and lignan glycosides from the twigs and leaves of an endangered conifer, Cathaya argyrophylla.

Labdane diterpene glycosides cathargyroside A and cathargyroside B, monoterpene glycosides vervenone-10-O-ß-D-glucopyranoside and vervenone-10-O-ß-D-apiofuranosyl-(1″→6')-ß-D-glucopyranoside, as well as lignan glycosides cedrusinin-4-O-α-L-rhamnopyranoside and (+)-cyclo-olivil-9'-O-ß-D-xylopyranoside, along with 39 known compounds, were obtained from the methanol extract of the twigs and leaves of Cathaya argyrophylla. These compounds were identified mainly by analyzing their NMR and MS data. Almost all of these compounds were hitherto unknown in this genus. The isolated compounds were screened against Candida albicans and Staphylococcus aureus for antimicrobial assay, and against K562, HT-29, BEL-7402, SGC-7901, B16, BGC-823, U251 and A549 cancer cell lines for cytotoxic activities. One compound showed antimicrobial activity against C. albicans, and four of them displayed cytotoxicity. Similarity analysis on the chemical constituents of the genera Cathaya, Picea and Pinus supported their close phylogenetic relationships.

[Determination of brodifacoum in rat plasma by HPLC].

OBJECTIVE: A determination method of brodifacoum in rat plasma with bromadiolone as an internal standard was developed. METHODS: A volume of 10 microl internal standard (bromadiolone) was added into rat plasma, and then extracted by 0.5 ml of acetonitrile by shaking for 2 min. The residue was dissolved with 200 microl of mobile phase after centrifugation for 10 min, and evaporation to dryness by Nitrogen blowing. A C18 column and PDA detector were used for separating and detecting. The wavelength was 254 nm, the flow rate was 1.0 ml/min, and the injection volume was 20 microl. RESULTS: The liner range was 1.0-20 microg/ml, and the correlation coefficient was 0.9992. The detection limit was 0.3 microg/ml in plasma (S/N=3). The intra-assay and inter-assay coefficients of variation were 1.89%-2.45% and 2.51%-3.61% respectively. The recoveries in plasma at levels of low, middle and high concentrations were (80.8 +/- 3.1)%, (81.8 +/- 2.7)% and (87.9 +/- 3.6)% (n=6), respectively. The accuracies were 84.1%-91.5% and 86.7%-93.2%, respectively. CONCLUSION: This method is simple, fast and accurate for the determination of brodifacoum in rat plasma.

Antimicrobial, cytotoxic lignans and terpenoids from the twigs of Pseudolarix kaempferi.

Seven new compounds, including four lignans, (+)-(8S,8'S)-9,9'-dibenzoylsecoisolariciresinol (1), (+)-(8S*,8'R*)-4,4'-dimethyloxomatairesinol (2), (+)-(7S*,8R*,8'R*,9'S*)-9'-n-butoxytsugacetal (3), and pseudolarkaemin A (4), a pyronane glycoside, pseudolarkaemin B (5), an ent-beyerene glycoside, pseudolarkaemin C (6), and a triterpene, 25-epi-pseudolarolide Q (7), along with 25 known compounds (8­32) were isolated from the twigs of Pseudolarix kaempferi. Their structures were elucidated mainly by the analysis of their NMR and MS data. Pseudolarolide C acid (24) was isolated for the first time as a natural product. All compounds were evaluated for antimicrobial activity against Candida albicans and Staphylococcus aureus, and cytotoxic activity against K562, HT-29, B16, BGC-823, BEL-7402, SGC-7901, U251, and A549 cancer cell lines were assayed. Results indicated that the new compounds 3, 7, and some known compounds showed antimicrobial and cytotoxic activities.

Rapid determination of aflatoxins in corn and peanuts.

A rapid and simple method using ultra-high-pressure liquid chromatography with UV detection for the determination of aflatoxins B1, B2, G1 and G2 in corn and peanuts has been developed. In this method, aflatoxins were extracted with a mixture of acetonitrile and water (86:14) and then purified by solid-phase clean-up with a MycoSep#226 AflaZon(+) column. The toxins were determined by UPLC-UV without derivatizing aflatoxins in real samples, which has not been used in other studies. The mean recoveries of aflatoxins from non-infected peanut and corn samples spiked with aflatoxins B1, B2, G1 and G2 at concentrations from 0.22 to 5 microg/kg were between 83.4% and 94.7%. The detection limits (S/N=3) for B1, B2, G1 and G2 were 0.32, 0.19, 0.32 and 0.19 microg/kg, and the corresponding quantification limits (S/N=10) were 1.07, 0.63, 1.07 and 0.63 microg/kg, respectively. We also applied this method on real samples. Among 16 peanut samples, 2 (12.5% incidence) were contaminated with aflatoxin; among 18 corn samples, 4 (22% incidence) were contaminated. The proposed method is rapid, simple and accurate for monitoring aflatoxins in corn and peanuts.

[Determination of the concentration of ropivacaine in dog plasma by reversed-phase high performance liquid chromatography].

OBJECTIVE: To establish a Reversed-Phase high performance liquid chromatography method for determination of plasma ropivacaine in dog. METHODS: The concentration of ropivacaine was assayed on Intertsil C18 column (4.6 mm x 250 mm, 5 microm)with a mobile phase consisting of methanol: H2O (50:50), the detection wavelength was 215nm and the flow rate was 1.4 ml/min. the column temperature was 20 degrees C and the sensitivity was 0.02 AUFS respectively. the volume of sample for detection is 10 microl. RESULTS: the linear range was 0.1-25 microg/ml (r = 0.9982). The recovery of ropivacaine was 91.2%-93.6%, RSD were 2.10%-3.40% (n=6). The detection limit was 0.05 microg/ml. the intra-day and interday precisions of assay for ropivacaine was 1.35%-2.88% and 1.80%-3.76%. CONCLUSION: This method is simple, rapid, accurate and convenient for determination of the concentration of ropivacaine in dog plasma.