Effects of repeated afidopyropen treatment on the structure and function of the soil microbial community.

Chemical insecticides are the most effective pest control agents. Afidopyropen is a novel insecticide used against sap-sucking insects, such as aphids. However, the effects of repeated afidopyropen application on the structure and function of soil microorganisms remain unknown. In this study, the changes in the enzyme activities, community structure and function, and relative abundance of antibiotic resistance ontology (ARO) of soil microorganisms were investigated during three repeated afidopyropen applications under laboratory conditions at the maximum recommended dosage (M1) and 10 times the M1 (M10). The neutral phosphatase (NPA) and catalase (CAT) activities in the soil were significantly suppressed after afidopyropen treatment. The Simpson diversity index (1/D) and Shannon-Wiener diversity index (H) also decreased in both the M1 and M10 afidopyropen-treated soils, indicating a remarkable decrease in soil microorganism diversity. The average well color development (AWCD) first increased and subsequently recovered to normal levels after the third application of the insecticide, suggesting that afidopyropen application could increase the metabolic activity of soil microorganisms. Metagenomic analysis showed that repeated afidopyropen application in both the M1 and M10 treatment groups altered the community structure of soil microorganisms, albeit in different ways. Furthermore, repeated afidopyropen application significantly increased the relative ARO abundance, especially in the M10 treatment, with the most dominant AROs being adeF, baeS, and IND-6. These findings reveal the effects of excessive afidopyropen application on soil microorganisms and lay an important foundation for the comprehensive evaluation of the impact of this insecticide on the environment.

Improvement of Bemisia tabaci (Hemiptera: Aleyrodidae) Fitness on Chinese Kale upon Simultaneous Herbivory by Plutella xylostella (Lepidoptera: Plutellidae).

Bemisia tabaci and the diamondback moth (DBM), Plutella xylostella, are two major cosmopolitan pests that often occur together and cause severe economic losses to cruciferous crops. However, little is known about how they interact with each other. To determine the effects of defense responses induced by the two pests on the biology and population dynamics of the herbivores, we studied the performance and fitness of B. tabaci and DBM when they damaged Chinese kale simultaneously and in different orders. The results showed that DBM pre-infestation shortened the developmental duration, increased longevity, oviposition days, and fecundity of B. tabaci. Meanwhile, the intrinsic rate of increase (r), net reproductive rate (R0) and finite rate of increase (λ) of B. tabaci increased significantly with dual infection as compared with only B. tabaci infestation. In contrast, B. tabaci pre-infestation reduced the longevity and oviposition days of DBM, but the population parameters r, R0, and λ did not vary significantly compared with only DBM infestation. Thus, co-infestation of B. tabaci and DBM was beneficial to the performance of the B. tabaci population. The present findings highlight that B. tabaci has become a dominant competitor when mixing with DBM on the same host plant.

Identification of SARS-CoV-2 entry inhibitors among already approved drugs.

To discover effective drugs for COVID-19 treatment amongst already clinically approved drugs, we developed a high throughput screening assay for SARS-CoV-2 virus entry inhibitors using SARS2-S pseudotyped virus. An approved drug library of 1800 small molecular drugs was screened for SARS2 entry inhibitors and 15 active drugs were identified as specific SARS2-S pseudovirus entry inhibitors. Antiviral tests using native SARS-CoV-2 virus in Vero E6 cells confirmed that 7 of these drugs (clemastine, amiodarone, trimeprazine, bosutinib, toremifene, flupenthixol, and azelastine) significantly inhibited SARS2 replication, reducing supernatant viral RNA load with a promising level of activity. Three of the drugs were classified as histamine receptor antagonists with clemastine showing the strongest anti-SARS2 activity (EC50 = 0.95 ± 0.83 µM). Our work suggests that these 7 drugs could enter into further in vivo studies and clinical investigations for COVID-19 treatment.

Overview of therapeutic drug research for COVID-19 in China.

Since the outbreak of novel coronavirus pneumonia (COVID-19) in December 2019, more than 2,500,000 people worldwide have been diagnosed with SARS-CoV-2 as of April 22. In response to this epidemic, China has issued seven trial versions of diagnosis and treatment protocol for COVID-19. According to the information that we have collected so far, this article provides an overview of potential therapeutic drugs and compounds with much attention, including favipiravir and hydroxychloroquine, as well as traditional Chinese medicine, which have been reported with good clinical treatment effects. Moreover, with further understanding of SARS-CoV-2 virus, new drugs targeting specific SARS-CoV-2 viral components arise and investigations on these novel anti-SARS-CoV-2 agents are also reviewed.

Microbial quantitation of colostrum from healthy breastfeeding women and milk from mastitis patients.

BACKGROUND: Colostrum is rich in microbiota. However, the quantity of microorganisms including both opportunistic pathogens and commensal mammary microbiota remains fluctuant during lactation. And once dysbiosis occurred in these microorganisms, a process in which the population of opportunistic pathogens increases while other bacteria, commensal mammary microbiota decrease. Lactation mastitis might occur. There were few literatures of microbiota in Chinese breastfeeding women. So this study aimed to investigate the quantity of microbiota in the colostrum from healthy breastfeeding women and the milk from mastitis patients in China. METHODS: From January to December 2017, a total of 400 milk samples were collected from the bilateral breasts of 200 women (104 healthy women and 86 mastitis patients). Microbial quantitation was done based on the conserved marker gene 16s rRNA for Bifidobacterium, Lactobacillus, Staphylococcus and Streptococcus by Real-time PCR in all milk samples. The bacterial culture of milk from mastitis patients was also performed. RESULTS: In the colostrum, the amounts of Lactobacillus and Bifidobacterium were significantly higher than those of Staphylococcus and Streptococcus (P<0.0001). The amounts of all the detected bacteria in the colostrum were significantly higher than in the milk from mastitis patients (P<0.01). The same results were obtained in patients with bacteria unrelated mastitis (P<0.01). With respect to colostrum samples, the Staphylococcus copies increased and Bifidobacterium copies declined in cases caused by Staphylococcus aureus or Methicillinresistant Staphylococcus aureus, while both the Staphylococcus and Bifidobacterium copies declined in the milk from patients with Staphylococcus epidermidis or Staphylococcus Lentus induced mastitis (P<0.05). CONCLUSIONS: Results of this study reveal a large amount of microbiota in the colostrum, and mammary microbial dysbiosis may be involved in the pathogenesis of lactational mastitis.

Nutrition versus defense: Why Myzus persicae (green peach aphid) prefers and performs better on young leaves of cabbage.

Plant leaves of different ages differ in nutrients and toxic metabolites and thus exhibit various resistance levels against insect herbivores. However, little is known about the influence of leaf ontogeny on plant resistance to phloem-feeding insects. In this study, we found that the green peach aphid, Myzus persicae, preferred to settle on young cabbage leaves compared with mature or old leaves, although young leaves contained the highest concentration of glucosinolates. Furthermore, aphids feeding on young leaves had higher levels of glucosinolates in their body, but aphids performed better on young leaves in terms of body weight and population growth. Phloem sap of young leaves had higher amino acid:sugar molar ratio than mature leaves, and aphids feeding on young leaves showed two times longer phloem feeding time and five times more honeydew excretion than on other leaves. These results indicate that aphids acquired the highest amount of nutrients and defensive metabolites when feeding on young cabbage leaves that are strong natural plant sinks. Accordingly, we propose that aphids generally prefer to obtain more nutrition rather than avoiding host plant defense, and total amount of nutrition that aphids could obtain is significantly influenced by leaf ontogeny or source-sink status of feeding sites.

Protective effects of traditional Tibetan medicine Zuo-Mu-A Decoction () on the blood parameters and myocardium of high altitude polycythemia model rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To explore the protective effects of Tibetan medicine Zuo-Mu-A Decoction (, ZMAD) on the blood parameters and myocardium of high altitude polycythemia (HAPC) model rats.</p><p><b>METHODS</b>Forty male Wistar rats were randomly divided into 4 groups by a random number table, including the normal, model, Rhodiola rosea L. (RRL) and ZMAD groups (10 in each group). Every group was raised in Lhasa to create a HAPC model except the normal group. After modeling, rats in the RRL and the ZMAD groups were administered intragastrically with RRL (20 mL/kg) and ZMAD (7.5 mL/kg) once a day for 2 months, respectively; for the normal and the model groups, 5 mL of distilled water was administered intragastrically instead of decoction. Then routine blood and hematologic rheology parameters were taken, levels of erythropoietin and 8-hydroxy-2'-deoxyguanosine (8-OHdG) were tested, and ultrastructural change in the left ventricular myocardium was observed using transmission electron microscopy.</p><p><b>RESULTS</b>Compared with the model group, ZMAD significantly reduced the red blood cell count, hemoglobin levels, whole blood viscosity at low/middle shear rates, plasma viscosity, erythrocyte electrophoretic time, erythropoietin and 8-OHdG levels, and also increased the erythrocyte deformation index (P<0.05). There was no difference in all results between the RRL and the ZMAD groups. The cardiac muscle fibers were well-protected, mitochondrial matrix swelled mildly and ultrastructure changes were less prominent in the ZMAD group compared with the model group.</p><p><b>CONCLUSION</b>ZMAD has significant protective effects on the blood parameters against HAPC, and also has the beneficial effect in protecting against myocardial injury.</p>

Chinese tuina downregulates the elevated levels of tissue plasminogen activator in sciatic nerve injured Sprague-Dawley rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To elucidate the mechanism of Chinese tuina in treating sciatic nerve crush injury, and to detect the levels of tissue plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1), which is thought to play an important role in nerve regeneration.</p><p><b>METHODS</b>Thirty-two adult male Sprague-Dawley rats were subjected to sciatic nerve crush injury and 16 rats (sham-operated group) went through a sham operation. Control group was given no treatment while tuina group received tuina therapy since day 7 post-surgery. Tuina treatment was performed once a day and lasted for 20 days. The sciatic functional index was examined every 5 days during the treatment session. The rats' gastrocnemius muscles were evaluated for changes in mass and immunohistochemistry techniques were performed to detect the levels of tPA and PAI-1.</p><p><b>RESULTS</b>Tuina therapy improved the motor function of sciatic nerve injured rats (P<0.05), however, it did not increase muscle volume (P<0.05). Tuina downregulated the levels of tPA and PAI-1 (P<0.05).</p><p><b>CONCLUSIONS</b>The present study implies that tuina treatment could accelerate rehabilitation of peripheral nerve injury.</p>

Exogenous kallikrein protects against diabetic nephropathy.

The kallikrein-kinin system has been shown to be involved in the development of diabetic nephropathy, but specific mechanisms are not fully understood. Here, we determined the renal-protective role of exogenous pancreatic kallikrein in diabetic mice and studied potential mechanisms in db/db type 2 diabetic and streptozotocin-induced type 1 diabetic mice. After the onset of diabetes, mice were treated with either pancreatic kallikrein (db/db+kallikrein, streptozotocin+kallikrein) or saline (db/db+saline, streptozotocin+saline) for 16 weeks, while another group of streptozotocin-induced diabetic mice received the same treatment after onset of albuminuria (streptozotocin'+kallikrein, streptozotocin'+saline). Db/m littermates or wild type mice were used as non-diabetic controls. Pancreatic kallikrein had no effects on body weight, blood glucose and blood pressure, but significantly reduced albuminuria among all three groups. Pathological analysis showed that exogenous kallikrein decreased the thickness of the glomerular basement membrane, protected against the effacement of foot process, the loss of endothelial fenestrae, and prevented the loss of podocytes in diabetic mice. Renal fibrosis, inflammation and oxidative stress were reduced in kallikrein-treated mice compared to diabetic controls. The expression of kininogen1, tissue kallikrein, kinin B1 and B2 receptors were all increased in the kallikrein-treated compared to saline-treated mice. Thus, exogenous pancreatic kallikrein both prevented and ameliorated diabetic nephropathy, which may be mediated by activating the kallikrein-kinin system.

Discovering novel anti-HCV compounds with inhibitory activities toward HCV NS3/4A protease.

AIM: To discover novel hepatitis C virus (HCV) inhibitors and elucidate the mechanism of action of the active compounds. METHODS: HCV subgenomic replicon-based luciferase reporter cell line was used to screen 1200 synthetic compounds with novel structures. Huh7.5.1 cell line stably transfected with HCV NS3/4A protease reporter was established to investigate the anti-HCV mechanism of the active compounds. The active compounds were further examined in an in vitro HCV infection assay to confirm their anti-HCV activity. RESULTS: After two-round screening in the anti-HCV replicon assay, some 2,4-diaminoquinazoline derivatives and carboxamide analogues were found to possess anti-HCV replicon activities (the IC50 values were less than 5 µmol/L). Among them, two representative compounds HZ-1157 and LZ-110618-6 inhibited HCV NS3/4A protease with IC50 values of 1.0 and 0.68 µmol/L, respectively. Furthermore, HZ-1157 and LZ-110618-6 inhibited HCV infection in vitro with IC50 values of 0.82 and 0.11 µmol/L, respectively. CONCLUSION: Some 2,4-diaminoquinazoline derivatives and carboxamide analogues have been identified as novel anti-HCV compounds.

Risk assessment of selected insecticides on Tamarixia triozae (Hymenoptera: Eulophidae), a parasitoid of Bactericera cockerelli (Hemiptera: Trizoidae).

Tamarixia triozae (Burks) (Hymenoptera: Eulophidae) is an important parasitoid of the potato or tomato psyllid, Bactericera cockerelli (Sulc) (Hemiptera: Trizoidae), a serious pest of potato (Solanum tuberosum L.), tomato (Solanum lycopersicum L.), and other solanaceous vegetables in many countries. To produce a marketable crop, insecticides are required when B. cockerelli populations reach economically damaging levels. We evaluated 11 commonly used insecticides for their effects on T. triozae. Glass-surface residues of spinetoram, imidacloprid-cyfluthrin, abamectin, and tolfenpyrad caused 100% mortality of T. triozae in 72 h, and the leaf residue of spinetoram was extremely toxic to T. triozae adults; even 15-d-old residues caused 100% mortality. Cyantraniliprole, fenpyroximate, pymetrozine, spirotetramat, spiromesifen, and chenopodium oil did not cause significant mortality in either glass surface or leaf-residue bioassays. Ingestion of spinetoram, abamectin, and imidacloprid+cyfluthrin (Leverage) by the adults resulted in 100% mortality in 12 h, and tolfenpyrad, 75.0% mortality in 12 h; whereas chenopodium oil and pymetrozine showed moderate effects on adult survival. Ingestion of abamectin, imidacloprid-cyfluthrin, and spinetoram killed all adults in the first day of treatment, whereas female adults in the treatment of pymetrozine lived 80.8 d, which was similar to those in the control. Ingestion of abamectin, imidacloprid-cyfluthrin, chenopodium oil, and spinetoram killed all male adults in the first day, whereas ingestion of other insecticides did not cause significant mortality, but reduced percent parasitism. Abamectin, imidacloprid-cyfluthrin, and spinetoram had the most deleterious effects on T. triozae, and have the least potential for use in integrated control programs using this parasitoid.

WSS45, a sulfated alpha-D-glucan, strongly interferes with Dengue 2 virus infection in vitro.

AIM: To investigate the mode of action of WSS45, one sulfated derivative of an alpha-D-glucan from the Gastrodia elata Bl, on the multiplication cycle of dengue virus serotype 2 (DV2), including initial infection and intracellular replication. METHODS: Virus multiplication in BHK cells were monitored by qRT-PCR, plaque reduction assay, and flow cytometry. Initial virus infection was dissected into adsorption and penetration steps by converting temperature and treating by acid glycine. Surface bound virions were detected by immunofluorescence staining for Evelope protein. RESULTS: WSS45 effectively inhibited DV2 infection in BHK cells with an EC(50) value of 0.68+/-0.17 microg/mL, mainly interfered with virus adsorption, in a very early stage of the virus cycle. However, WSS45 showed no viricidal effect. Moreover, WSS45 could increase the detaching of virus from cell surface in BHK cell line. CONCLUSION: WSS45 exerted potent inhibitory effect on DV2 through interfering with the interaction between viruses and targeted cells. This activity was related to its molecular size.

[Color Doppler ultrasound evaluation of a Chinese herbal formula in treating lower limb arteriosclerosis: a randomized controlled trial].

OBJECTIVE: To observe the effects of a Chinese compound herbal medicine for reinforcing qi, activating blood circulation, expelling phlegm and dredging collaterals in treatment of lower limb arteriosclerosis. METHODS: The outpatients and inpatients with lower limb arteriosclerosis from Shanghai Yueyang Hospital of Integrated Traditional Chinese and Western Medicine were randomly divided into control group and treatment group after color Doppler ultrasound inspection, and there were 30 cases in each group. The patients in the treatment group were treated with simvastatin and a Chinese compound herbal medicine for reinforcing qi, activating blood circulation, expelling phlegm and dredging collaterals, and the patients in the control group were treated only with simvastatin. The color Doppler ultrasound imaging of lower limb arteries including inside diameter, intima-media thickness, and plaque area in all the patients were examined after 6-month treatment. The levels of serum total cholesterol (TC) and low-density lipoprotein (LDL) were detected too. RESULTS: After the treatment, the lower limb artery intima-media thickness and artery plaque area in the treatment group decreased obviously as compared with those in the control group (P < 0.05). Color Doppler flow imaging showed that the percentage of patients with uniphase wave decreased from 26.67% (8/30) to 6.67% (2/30) in the treatment group (P < 0.05), and from 26.67% (8/30) to 16.67% (5/30) in the control group (P < 0.05) as compared with those before the treatment, and there was also a significant difference between the two groups (P < 0.05). The concentrations of serum TC and LDL in the treatment group were lower than those in the control group (P < 0.05). CONCLUSION: Color Doppler ultrasound showed that the Chinese compound medicine for reinforcing qi, activating blood circulation, expelling phlegm and dredging collaterals has positive effects on lower limb arteriosclerosis.

Experience-altered oviposition responses to a neem-based product, Neemix, by the diamondback moth, Plutella xylostella.

The oviposition responses of the diamondback moth, Plutella xylostella L., to Neemix 4.5, a neem-based oviposition repellent/deterrent, with or without previous experience were studied in the laboratory on plain aluminum foil sheets, on aluminum foil sheets coated with cabbage juice, and on cabbage plants. In the plain aluminum foil sheet experiment, the females without prior experience of Neemix (inexperienced females) deposited more eggs (66.2%) on sections with untreated sheets than on sections with Neemix-treated sheets (33.8%), indicating that the inexperienced females were significantly repelled and/or deterred by Neemix. In contrast, the Neemix-experienced females deposited more eggs (69.2-69.7%) on sections with Neemix-treated sheets than on sections with untreated sheets (30.3-30.8%), implying that the females were not repelled and/or deterred by Neemix following an experience. In the tests using egg-laying sheets coated with cabbage juice or with cabbage juice plus Neemix, the inexperienced females deposited more eggs (63.8%) on the sections with sheets treated with cabbage juice only than on the sections with sheets coated with cabbage juice and Neemix (36.2%), indicating that inexperienced females were repelled and/or deterred by Neemix. Similarly, Neemix-experienced females deposited more eggs (62.2-65.6%) on the sections with sheets treated with cabbage juice plus Neemix than on the sections with sheets treated with juice only (34.4-37.8%), suggesting that the females were not repelled and/or deterred by Neemix following an experience. On cabbage plants, inexperienced females oviposited similar numbers of eggs on both Neemix-treated (50.8%) and untreated plants (49.2%). However, the Neemix-experienced females oviposited more eggs on the plants treated with Neemix (56.3%) than on untreated plants (43.7%), indicating that the females were attracted by Neemix-treated plants following an experience. The significance of this study for applications of insect repellents/deterrents in pest management is discussed.