Preliminary Exploration of Potential Active Ingredients and Molecular Mechanisms of Yanggan Yishui Granules for Treating Hypertensive Nephropathy Using UPLC-Q-TOF/MS Coupled with Network Pharmacology and Molecular Docking Strategy.

Hypertensive nephropathy (HN) is a prevalent complication of hypertension and stands as the second primary reason for end-stage renal disease. Research in clinical settings has revealed that Yanggan Yishui Granule (YGYSG) has significant therapeutic effects on HN. However, the material basis and action mechanisms of YGYSG against HN remain unclear. Consequently, this study utilized a comprehensive method integrating ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS), network pharmacology, and molecular docking to delineate the active ingredients and potential therapeutic mechanisms of YGYSG for treating HN. Firstly, sixty distinct components were recognized in total as potential active ingredients in YGYSG by UPLC-Q-TOF/MS. Subsequently, the mechanisms of YGYSG against HN were revealed for the first time using network pharmacology. 23 ingredients played key roles in the complete network and were the key active ingredients, which could affect the renin-angiotensin system, fluid shear stress and atherosclerosis, HIF-1 signaling pathway, and AGE-RAGE signaling pathway in diabetic complications by regulating 29 key targets such as TNF, IL6, ALB, EGFR, ACE, and MMP2. YGYSG could treat HN through the suppression of inflammatory response and oxidative stress, attenuating the proliferation of renal vascular smooth muscle cells, lessening glomerular capillary systolic pressure, and ameliorating renal dysfunction and vascular damage through the aforementioned targets and pathways. Molecular docking results revealed that most key active ingredients exhibited a high affinity for binding to the key targets. This study pioneers in clarifying the bioactive compounds and molecular mechanisms of YGYSG against HN and offers scientific reference into the clinical application.

Host relationships and biological notes of Cassidinae beetles (Coleoptera, Chrysomelidae) in Qiannan Prefecture, Guizhou, China.

The faunal composition, host relationships and biological information of the subfamily Cassidinae (Coleoptera, Chrysomelidae) remain poorly known in many Chinese regions. Based on the seven-year field survey, faunal composition and host associations of Cassidinae beetles were systematically compiled for Qiannan Buyi and Miao Autonomous Prefecture, Guizhou Province. In particular, through direct field observations, detailed biological information, such as life history and behavioural features and host plants were first recorded for 56 species of Cassidinae beetles. We have tripled the number of Cassidinae species in Qiannan. Sixty-nine species of Cassidinae beetles belonging to 17 genera and eight tribes were identified, of which 38 species are newly recorded in Guizhou and 56 are newly recorded in Qiannan. The tribes Leptispini and Notosacanthini were newly recorded in Guizhou. The genera Thlaspidosoma Spaeth, Downesia Baly, Klitispa Uhmann, Platypria Guérin-Méneville, Leptispa Baly and Notosacantha Chevrolat were recorded in Guizhou for the first time. A total of 61 species, 37 genera and 17 families of host plants were collected. Lardizabalaceae and Araliaceae were new host plant families for Cassidinae worldwide. Quantitative food web analysis indicated that Cassidinae species in Qiannan mainly feed on Poaceae, Rosaceae, Convolvulaceae and Lamiaceae. Callispini and Leptispini only feed on monocots, Aspidimorphini, Basiprionotini, Cassidini and Notosacanthini only feed on dicots, while Hispini feed on both monocots and dicots. The feeding patterns and corresponding damage marks of Cassidinae were quite diverse. In addition, the pupal mine-making behaviour of Dactylispaexcisa (Kraatz, 1879), D.similis Chen et T'an, 1985 and D.uhmanni Gressitt, 1950 are worth further study. Although preliminary, our field survey is an essential step in understanding Cassidinae behaviour and Cassidinae-plant interactions.

Culex quinquefasciatus membrane-bound alkaline phosphatase is a putative receptor for Lysinibacillus sphaericus Tpp49Aa1 toxin.

The binary toxin Cry48Aa1/Tpp49Aa1 produced by Lysinibacillus sphaericus exhibits potent toxicity against Culicidae larvae. Both Cry48Aa1 and Tpp49Aa1 toxins are crucial for binding to the toxin receptor in Culex quinquefasciatus larvae, albeit with different binding sites. Previous studies have identified Glu71, a membrane-bound α-glucosidase, as a putative binding protein for the Cry48Aa1 toxin, involved in the Cry48Aa1/Tpp49Aa1 toxicity. In this study, we employed pulldown assays to identify a group of Tpp49Aa1-binding proteins from C. quinquefasciatus solubilized midgut brush-border membrane proteins (BBMFs). RNA interference assays revealed that the silencing of an alkaline phosphatase gene (referred to as ALP1263) in C. quinquefasciatus resulted in a significant reduction in larval mortality upon exposure to Cry48Aa1/Tpp49Aa1 toxin in vivo. Furthermore, the ALP1263 protein exhibited specific and high-affinity binding to the Tpp49Aa1 toxin, with a dissociation constant (Kd) of approximately 57.3 nM. The dot blot analysis demonstrated that Tpp49Aa1 C-terminal region was essential for its interaction with the ALP1263 protein. In summary, our findings establish ALP1263 as a functional receptor for Tpp49Aa1 and emphasize its role in the toxicity of Cry48Aa1/Tpp49Aa1.

Field survey of Cassidinae beetles (Coleoptera, Chrysomelidae) and their host plants in southern Guangxi, China.

Few systematic studies have been conducted on the faunal composition and food web structure of Cassidinae of China. During 2013-2019, we systematically investigated Cassidinae beetles and their host plants in the southern Guangxi. A total of 2,255 Cassidinae individuals from 66 species, 23 genera and ten tribes were collected in southern Guangxi. Most species belonged to the tribe Hispini (23 species, 34.8%), followed by the tribe Gonophorini (13 species, 19.7%), Cassidini (eight species, 12.1%) and Aspidimorphini (six species, 9.1%). The others (16 species) belonged to the tribes Anisoderini, Botryonopini, Callispini, Oncocephalini, Notosacanthini and Leptispini. The tribe Notosacanthini was recorded from Guangxi for the first time. The genera Neownesia (Botryonopini), Gonophora (Gonophorini), Micrispa (Gonophorini), Notosacantha (Notosacanthini) and Prionispa (Oncocephalini) were firstly recorded in Guangxi. In total, we obtained 47 newly-recorded species in southern Guangxi and 33 newly-recorded species in the whole Guangxi, of which, Callispafrontalis Medvedev, 1992 was newly recorded in China. Dactylispafeae Gestro (625 individuals) and D.chinensis Weise (565 individuals) were the most common species. A total of 69 species, 53 genera and 19 families of host plants were identified for Cassidinae in southern Guangxi. Many host plant associations are new records for Cassidinae. Quantitative food web analysis indicated that Cassidinae species in southern Guangxi primarily fed on Poaceae, Convolvulaceae, Cyperaceae and Rosaceae. Generally, the plant-Cassidinae food webs were moderately complex and stable in southern Guangxi. This is the first large contribution to the knowledge of the species composition and host plant diversity of Cassidinae in southern Guangxi.

Association between salt sensitivity and blood pressure variability in patients with essential hypertension and predictive value for cardiovascular events.

We aimed to explore the association between salt sensitivity and blood pressure variability in patients with essential hypertension. A total of 730 patients with essential hypertension treated from 2016 to 2019 were subjected to salt-sensitivity risk stratification according to 24-hour ambulatory blood pressure monitoring. Their clinical data were compared among groups with different grades of salt-sensitivity risk, and the association between salt sensitivity and blood pressure variability was analysed. The influencing factors for cardiovascular events in patients with essential hypertension were analysed through multivariate regression analysis, and their predictive value was detected using receiver operating characteristic (ROC) curves. Salt sensitivity was positively correlated with night-time and 24-hour systolic standard deviation and 24-hour systolic blood pressure coefficient of variation. Age ≥ 55 years, family history of cardiovascular diseases, high risk of salt sensitivity, night-time systolic standard deviation ≥ 14 mmHg, 24-hour systolic standard deviation ≥ 20 mmHg and 24-hour systolic blood pressure coefficient of variation ≥ 13.5% were all independent risk factors for cardiovascular diseases in patients with essential hypertension (p < 0.05). The area under the ROC curve of the prediction model was 0.837. There was a positive correlation between salt sensitivity and blood pressure variability, which has predictive value for cardiovascular events in patients with essential hypertension.

Genera of Tischeriidae (Lepidoptera): a review of the global fauna, with descriptions of new taxa.

For the first time, genera of leaf mining Tischeriidae of the global fauna are reviewed and four new genera are described: Coptotrichoides Dikus & Stonis, gen. nov., trophically associated with Sapindaceae from Central and South America; Rytietia Dikus, Xu & Dai, gen. nov., trophically associated with Annonaceae from East and South East Asia; Pafazaria Dikus & Stonis, gen. nov., trophically associated with Fabaceae and Malvaceae from East and South East Asia; and Gnathitischeria Dikus, gen. nov., trophically associated with Malvaceae and Asteraceae from Central America. In addition to the descriptions of new genera, all previously described genera (Coptotriche Walsingham, 1890; Dishkeya Stonis, 2020; Tischeria Zeller, 1839; Manitischeria Dikus & Stonis, 2021; Neotischeria Dikus & Stonis, 2021; Paratischeria Dikus & Stonis, 2017; and Astrotischeria Puplesis & Dikus, 2003) are characterized and a pictorial identification key for all 11 genera is provided. Morphological and biological characteristics of all genera are supplemented with some molecular data. We described 12 new species: South East Asian Coptotriche sapaensis Dikus & Stonis, sp. nov.; Central American Coptotrichoides sapindoidum Dikus & Stonis, sp. nov. and Tischeria neokristenseni Dikus & Stonis, sp. nov.; South East Asian Rytietia uncinata Dikus, Xu & Dai, sp. nov., East Asian R. chongyiensis Xu & Dai, sp. nov.; South East Asian Pafazaria capitata Dikus & Stonis, sp. nov. and P. faboidica Dikus & Stonis, sp. nov.; Central American Neotischeria antigua Dikus & Remeikis, sp. nov. and N. subantigua Dikus & Remeikis, sp. nov.; South American Astrotischeria incae Dikus & Stonis, sp. nov.; and Central American Gnathitischeria atitlani Dikus & Stonis, sp. nov. and G. arcana Dikus & Stonis, sp. nov. The taxonomic rank of the East Asian subspecies Tischeria decidua siorkionla Kozlov, 1986 is elevated to species rank: T. siorkionla Kozlov (stat. nov.). The male genitalia of holotypes, paratypes, lectotypes or paralectotypes, or non-type specimens of some little-known species are examined for the first time. We provide the first morphological documentation for the following: Tischeria ceanothi lectotype and paralectotype; T. immaculata (stat. restored) paratype; Coptotrichoides deliquescens (Meyrick) lectotype and paralectotype; lectotypes of Coptotriche aenea (Frey & Boll), Astrotischeria plagifera (Meyrick), A. helianthi (Frey & Boll), and A. ephaptis (Meyrick); Coptotriche berberella (De Prins) paratype; and the first photographic documentation of Tischeria lvoskyi Kozlov holotype. Some non-type specimens deposited at NHMUK, including a few previously neglected species, are also examined and documented, including the American Astrotischeria solidagonifoliella (Clemens) and A. omissa (Braun). Nine new combinations are provided: Coptotrichoides deliquescens (Meyrick, 1915), comb. nov.; C. singularis (Stonis & Dikus) comb. nov.; C. suprafasciata (Dikus & Stonis) comb. nov., C. serjaniphaga (Remeikis & Stonis) comb. nov.; C. braziliensis (Dikus & Stonis) comb. nov.; Pafazaria jingdongensis (Xu & Dai), comb. nov.; Neotischeria explosa (Braun) comb. nov.; N. pallidipennella (Braun) comb. nov.; and Astrotischeria heteroterae (Frey & Boll) comb. nov. The synonymization of Tischeria longeciliata Frey & Boll, 1878 with Astrotischeria helianthi (Frey & Boll, 1878) is confirmed. One new species group, the Tischeria ceanothi group, is established. We also report a new distribution record of T. dodonaea Stainton from the Caucasus, Armenia. Three tables and 569 figures of adult external morphology, male and female genitalia, leaf mines, and cladograms based on molecular characters are provided.

First biological report on the genus Cantonius (Buprestidae, Agrilinae, Aphanisticini), with descriptions of two new species from China.

Background: The genus Cantonius Théry, 1929 is a small group with two subgenera and 12 species. However, the biology of this genus is still unknown. New information: In this paper, three species of the genus Cantonius Théry, 1929 were found on bamboo leaves, revealing for the first time that Cantonius species are also leaf-miners. Two new species were recorded from Jiangxi Province and are described here: Cantonius (Cantonius) anjiensis sp. n. (host plant: Pleioblastusamarus) and Cantonius (Procantonius) qiyunensis sp. n. (host plant: Bambusablumeana) followed by C. (P.) austrisinicus Kalashian, 2021 (host plant: Oligostachyumpaniculatum) recorded from Guangxi Province. Including habitats, photos of three species together with C. (P.) qiyunensis sp. n. pupa, host plants, and leaf mines of the three species are presented. Moreover, the bionomics and habits of the genus are discussed for the first time, and a hypothesis for the distribution of Cantonius is provided.

Hypoglossal nerve stimulation in adolescents with down syndrome and obstructive sleep apnea: A systematic review and meta-analysis.

Objective: To evaluate the efficacy and adverse effects of hypoglossal nerve stimulation in adolescents with down syndrome and obstructive sleep apnea. Methods: A systematic search was conducted using PubMed, Web of Science, Embase, and Scopus databases. The systematic review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. A comprehensive search strategy used a combination of Medical Subject Headings and free words with "OR" and "AND." Articles were screened to extract data reporting apnea-hypopnea index, quality of life, voltage, follow-up duration, and complications. All included participants were adolescents with down syndrome and obstructive sleep apnea. Results: A total of 92 articles were identified, of which 9 articles met the inclusion criteria. A total of 106 patients were included. All the studies showed that patients receiving hypoglossal nerve stimulation experienced a significant decrease in apnea-hypopnea index (at least 50%). The pooled AHI was significantly lower in patients following treatment (mean AHI reduction 17.43 events/h, 95% confidence interval 13.98-20.88 events/h, P < 0.001) after 2 case reports were excluded. The pooled OSA-18 were significantly decreased in 88 patients after treatment (mean OSA-18 reduction 1.67, 95% confidence interval 1.27-2.08, P < 0.001) after excluding 5 studies. Four investigations examined the necessity to optimize stimulation voltage for arousal during treatment. The most common complication was pain or discomfort in the tongue or mouth. Most studies had relatively short patient follow-up periods, with the most extended follow-up being 44-58 months. Conclusion: Hypoglossal nerve stimulation significantly reduces apnea-hypopnea index and improves the quality of life; and thus, could be a potential alternative therapy for obstructive sleep apnea in adolescents with down syndrome. The adolescent's age, potential complications, adverse events, long-term efficacy, and comfort, needs to be considered while performing hypoglossal nerve stimulation.

Application of the ratio of the radiopaque calcified area to the dental follicle (RCA/DF) for dental age assessment on orthopantomograms.

OBJECTIVES: This study aimed at exploring a new tooth development evaluation method for age assessment and investigating the dynamic alteration and potential trend of tooth development by orthopantomograms (OPGs), in order to provide references for tooth development prediction and forensic purpose. MATERIALS AND METHODS: A total of 132 OPGs of children aged 3-8 years were collected. The developmental stages of the permanent mandibular second molar (M2)were evaluated by experienced examiners according to the Nolla method and Mimics software, respectively. Quantitative analysis of the ratio of the radiopaque calcified area to the dental follicle (RCA/DF) in different stages, ages, sexes and quadrants were evaluated and compared via descriptive statistics and Spearman's correlation coefficient analysis. RESULTS: There was a strong, positive correlation between the examiners' evaluation and mimics analysis results. With the age increased, the Nolla stage of M2 observed by OPGs increased, and the RCA/DF showed increased trends both in males and females. There were significant differences of the RCA/DF of the M2 at various ages. The tooth calcification development of female was 9.08% earlier than that of male between 3 and 8 years old. However, teeth of male seemed to develop faster than that of female during this period. There was no significant difference between left and right quadrant either according to the Nolla stage or RCA/DF. CONCLUSIONS: The RCA/DF value obtained from OPGs of the developmental mandibular second permanent molars could be used as a reliable indicator for tooth maturity and age estimation in children aged 3-8 years. CLINICAL RELEVANCE: Age assessment based on radiographs is considered as a reliable and efficient indicator for judging different types of malocclusion, making suitable orthodontic treatment plan, deciding the extraction time of retained deciduous teeth in clinic practice.

An amelogenin-based peptide hydrogel promoted the odontogenic differentiation of human dental pulp cells.

Amelogenin can induce odontogenic differentiation of human dental pulp cells (HDPCs), which has great potential and advantages in dentine-pulp complex regeneration. However, the unstability of amelogenin limits its further application. This study constructed amelogenin self-assembling peptide hydrogels (L-gel or D-gel) by heating-cooling technique, investigated the effects of these hydrogels on the odontogenic differentiation of HDPCs and explored the underneath mechanism. The critical aggregation concentration, conformation, morphology, mechanical property and biological stability of the hydrogels were characterized, respectively. The effects of the hydrogels on the odontogenic differentiation of HDPCs were evaluated via alkaline phosphatase activity measurement, quantitative reverse transcription polymerase chain reaction, western blot, Alizarin red staining and scanning electron microscope. The mechanism was explored via signaling pathway experiments. Results showed that both the L-gel and D-gel stimulated the odontogenic differentiation of HDPCs on both Day 7 and Day 14, while the D-gel showed the highest enhancement effects. Meanwhile, the D-gel promoted calcium accumulation and mineralized matrix deposition on Day 21. The D-gel activated MAPK-ERK1/2 pathways in HDPCs and induced the odontogenic differentiation via ERK1/2 and transforming growth factor/smad pathways. Overall, our study demonstrated that the amelogenin peptide hydrogel stimulated the odontogenic differentiation and enhanced mineralization, which held big potential in the dentine-pulp complex regeneration.

Culex quinquefasciatus alpha-glucosidase serves as a putative receptor of the Cry48Aa toxin from Lysinibacillus sphaericus.

The Cry48Aa/Cry49Aa toxin of Lysinibacillus sphaericus shows specific toxicity towards larvae of Culex spp. Individual Cry48Aa and Cry49Aa subunits interact with distinct target sites in the larval midgut and overcome the resistance of Culex to the Bin toxin. However, the toxin-binding proteins have not yet been identified. The present study aimed to identify Cry48Aa-binding proteins in Culex quinquefasciatus. Pulldown assays using C. quinquefasciatus midgut brush-border membrane fractions (BBMFs) identified a class of proteins, including aminopeptidases (APNs), protease m1 zinc metalloproteases, alkaline phosphatases (ALPs), and maltases, that could be potentially involved in the mode of action of this toxin. RNA interference analysis showed that silenced larvae treated with dsRNA of the alpha-glucosidase (named Glu71) gene were more tolerant of the Cry48Aa/Cry49Aa toxin, which induced less than 20% mortality. The amino acid sequence of Glu71 exhibited 42% identity with Cqm1/Cpm1, which acted as a Bin toxin receptor. Toxin binding assays showed that Cry48Aa had a high specific binding capacity for the Glu71 protein, whereas Cry49Aa exhibited no specific binding. Overall, our results showed that Glu71 is a Cry48-binding protein involved in Cry48Aa/Cry49Aa toxicity.

Moxibustion alleviates chronic heart failure by regulating mitochondrial dynamics and inhibiting autophagy.

Moxibustion (MOX) is a traditional Chinese medicine preparation, which has been clinically used to treat cardiac diseases in recent years. The present study aimed to examine the protective effects and possible mechanisms of MOX on doxorubicin (DOX)-induced chronic heart failure (CHF) in rats. The animals were divided into five groups, including the Control (normal saline), DOX (doxorubicin 15 mg/kg), MOX (doxorubicin 15 mg/kg + moxibustion), BEN (doxorubicin 15 mg/kg + benazepril 0.86 mg/kg) and MOX + BEN (doxorubicin 15 mg/kg + moxibustion + benazepril 0.86 mg/kg) groups. After three weeks, echocardiography was performed to assess cardiac function and structure, including left ventricular internal diameter in systole, ejection fraction and fractional shortening (FS). Serum brain natriuretic peptide levels and adenosine triphosphate (ATP) levels were measured by enzyme-linked immunosorbent assay and ATP assay. Cardiac pathology was assessed by hematoxylin and eosin and Masson's trichrome staining. Cardiac ultrastructure and the number of autophagosomes formed were visualized by transmission electron microscopy. Western blotting was performed to assess mitochondrial dynamics, autophagy proteins and mitochondrial autophagy-related pathway proteins. The expression levels of these genes were assessed by reverse transcription-quantitative PCR. The results indicated MOX could improve cardiac function, increased cardiac ATP levels and reduced myocardial fibrosis. Western blotting indicated that MOX treatment elevated the expression of optic atrophy 1 protein (OPA1), while decreasing the expression of dynamin-related protein 1 and mitochondrial fission 1 protein. In addition, MOX inhibited autophagy, as evidenced by decreased number of autophagosomes, reduced LC3II/LC3I ratio and increased p62 expression. Furthermore, MOX downregulated DOX-induced FUNDC1 signaling pathway. In summary, MOX has protective effects on DOX-induced CHF in rats, promoting mitochondrial fusion while inhibiting mitochondrial fission and mitophagy. The underlying mechanisms may be related to the inhibition of the FUNDC1 signaling pathway.

Unveiling the Differences in Biological Properties of Dental Pulp Stem Cells from Normal and Inflamed Pulp: A Comprehensive Comparative Study.

BACKGROUND The aims of the study were to comprehensively compare the morphology, immunophenotype, proliferation, migration, and regeneration potential of normal dental pulp stem cells (DPSCs) versus inflammatory dental pulp stem cells (iDPSCs). MATERIAL AND METHODS Healthy pulp or inflamed pulp tissue was used to isolate and culture DPSCs and iDPSCs, respectively. These cell populations were characterized by flow cytometry, colony formation assay, transwell assay, and multi-directional differentiation in vitro. RESULTS No difference was observed in the morphology, cell-surface markers, or cell migration between DPSCs and iDPSCs. DPSCs showed a higher colony-forming capacity, proliferative viability, and osteo/dentinogenesis ability compared with iDPSCs. However, iDPSCs demonstrated enhanced neurogenesis, angiogenesis, adipogenesis, and chondrogenesis capacities in comparison to DPSCs. CONCLUSIONS Our data revealed the differences of biological properties between DPSCs and iDPSCs. The highly angiogenic and neurogenic potential of iDPSCs indicate their possible use in the regeneration of the dentin-pulp complex and support the critical role of angiogenesis and neurogenesis in pulp regeneration.

A checklist of gymnosperm-feeding leafminers (Arthopoda, Insecta) in North America and Europe.

The leafminers on gymnosperms receive much less attention than those on either angiosperms or ferns. Given the distinctly different leaf shape and leaf venation found in gymnosperms, they would be expected to host significantly different leafminer groups. Very few comprehensive reports on gymnosperm-feeding leafminers have been presented. Based on the well-studied fauna in North America and Europe, we compiled a list of 133 species, 30 genera and 13 families of gymnosperm-feeding leafminers. The gymnosperm-mining families (in descending order of leafminer number) included Tortricidae, Gelechiidae, Argyresthiidae, Yponomeutidae, Batrachedridae, Pyralidae, Adelidae, Agromyzidae, Blastobasidae, Bucculatricidae, Coleophoridae, Curculionidae and Noctuidae. There were 109 species, 22 genera and ten families in North America and 34 species, 19 genera and nine families in Europe. We compiled a list of 102 species and 16 genera of host plants, belonging to four families: Pinaceae, Cupressaceae, Taxaceae and Zamiaceae. There were 84 host species, 15 genera and three host families in North America and 46 host species, ten genera and three host families in Europe. Dominant gymnosperm-mining families and dominant host families were generally the same in the two continents.

Elevational Pattern of Leaf Mine Diversity on Quercus variabilis Blume at Baotianman, Henan, China.

The species composition and diversity pattern of leaf miners on dominant trees in China are poorly understood. Using Hill-based diversity metrics, the elevational patterns of taxonomic, phylogenetic, and functional diversity for leaf miners on Quercus variabilis Blume at Baotianman were systematically analyzed. Leaf mine types belonged to ten genera and seven families. Different leaf miners had different elevational preferences. Most taxonomic and phylogenetic Hill diversity indices had typical hump-shaped elevational patterns, with a peak at the middle elevation of approximately 875 m. No functional Hill diversity indices presented significant linear or nonlinear trends with altitude. The driving factors behind the elevational distribution patterns of leaf miners require further work.

Discovery of the new Coptotriche species in China revealed two novel host-plant families and host-plant orders for Tischeriidae, a family of stenophagous, leaf-mining lepidopterans.

The article describes three new species of Coptotriche Walsingham, 1890 from China and adjacent northern Vietnam: C. camptotheca Xu Dai, sp. nov. feeding on Camptotheca acuminata Decne. (Nyssaceae); C. turpinia Xu Dai, sp. nov. feeding on Turpinia arguta (Lindl.) Seem. (Staphyleaceae); and C. asiana Dikus Stonis, sp. nov. The latter appeared to be a narrow oligophagous species feeding on various Symplocos Jacq.: S. sumuntia Buch.-Ham. ex D. Don, S. poilanei Guill., and Symplocos glauca (Thunb.) Koidz. (Symplocaceae). Nyssaceae and Staphyleaceae are novel host-plant families of Tischeriidae. The new species are illustrated with photographs of adults, male and female genitalia, and leaf mines. The article also briefly discusses about the proportion of monophagous and oligophagous species of Tischeriidae.

Most trumpet moths dont feed on plants of the nettle family but Paratischeria does: the first discovery of Tischeriidae (Lepidoptera) on Urticaceae in Asia.

This is the first record of Urticaceae-feeding trumpet moths (Tischeriidae) from Asia. We describe Paratischeria boehmerica Dikus Stonis, sp. nov. and P. grossa Dikus Stonis, sp. nov., two distinctive new species of Tischeriidae recently discovered from Laos, South East Asia, feeding on plants of the nettle family. Urticaceae is a rare host-plant family for leaf-mining Tischeriidae worldwide. The new species are illustrated with photographs of the adults, male and female genitalia, and the leaf mines.

The First Eight Mitogenomes of Leaf-Mining Dactylispa Beetles (Coleoptera: Chrysomelidae: Cassidinae) Shed New Light on Subgenus Relationships.

The taxonomic classification of Dactylispa, a large genus of leaf-mining beetles, is problematic because it is currently based on morphology alone. Here, the first eight mitochondrial genomes of Dactylispa species, which were used to construct the first molecular phylogenies of this genus, are reported. The lengths of the eight mitogenomes range from 17,189 bp to 20,363 bp. All of the mitochondrial genomes include 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs), and 1 A + T-rich region. According to the nonsynonymous/synonymous mutation ratio (Ka/Ks) of all PCGs, the highest and the lowest evolutionary rates were found for atp8 and cox1, respectively, which is a common phenomenon among animals. According to relative synonymous codon usage, UUA(L) has the highest frequency. With two Gonophorini species as the outgroup, mitogenome-based phylogenetic trees of the eight Dactylispa species were constructed using maximum likelihood (ML) and Bayesian inference (BI) methods based on the PCGs, tRNAs, and rRNAs. Two DNA-based phylogenomic inferences and one protein-based phylogenomic inference support the delimitation of the subgenera Dactylispa s. str. and Platypriella as proposed in the system of Chen et al. (1986). However, the subgenus Triplispa is not recovered as monophyletic. The placement of Triplispa species requires further verification and testing with more species. We also found that both adult body shape and host plant relationship might explain the subgeneric relationships among Dactylispa beetles to a certain degree.

Effect of acidic culture conditions on the proliferation, apoptosis, and migration ability of human tongue squamous cell carcinoma cells and its related mechanism.

OBJECTIVES: This study aims to explore the effect of acidic culture conditions on the proliferation, apoptosis, and migration ability of human tongue squamous cell carcinoma SCC15 and CAL27 cells and its potential molecular mechanism. METHODS: After acidic culture for different periods, methyl thiazolyl tetrazolium (MTT) method was adop-ted to detect the cell proliferation of SCC15 and CAL27. Flow cytometry was employed to detect the apoptosis level of SCC15 and CAL27 cells. The migration ability of SCC15 and CAL27 after acidic culture was detected by scratch hea-ling test. Real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) was used to detect the mRNA expression of cyclooxygenase 2 (COX-2) and survivin in SCC15 and CAL27 cells after acidic culture. RESULTS: After culture for 24 h under acidic microenvironment, SCC15 and CAL27 cells grew rapidly and reached the stationary phase after adjustment for 3 days. The apoptosis levels of SCC15 and CAL27 cells decreased after acidic culture, but the most significant reduction occurred after 6 h of acidic culture. The scratch healing rates of SCC15 and CAL27 cells increased after acidic culture. The results of FQ-PCR showed that the mRNA expression levels of COX-2 and survivin in SCC15 and CAL27 cells increased after acidic culture. CONCLUSIONS: Extracellular acidic microenvironment can inhibit the apoptosis of tongue squamous carcinoma cells, promote their migration, and induce more adaptable and malignant tongue squamous carcinoma cells. The mechanism may be related to COX-2 and survivin and their signal pathways.

Cys183 and Cys258 in Cry49Aa toxin from Lysinibacillus sphaericus are essential for toxicity to Culex quinquefasciatus larvae.

The two-component Cry48Aa/Cry49Aa toxin produced by Lysinibacillus sphaericus shows specifically toxic to Culex quinquefasciatus mosquito larvae. Cry49Aa C-terminal domain is responsible for specific binding to the larval gut cell membrane, while its N-terminal domain is required for interaction with Cry48Aa. To investigate functional role of cysteine in Cry49Aa, four cysteine residues at positions 70, 91, 183, and 258 were substituted by alanine. All mutants showed similar crystalline morphology and comparable yield to that of the wild type except that the yield of the C91A mutant was low. Four cysteine residues did not involve in disulfide bond formation within or between Cry49Aa molecules. Cys91, Cys183, and Cys258 are essential for larvicidal activity against C. quinquefasciatus larvae, while Cys70 is not. Substitution at C91, C183, and C258 caused weaker Cry48Aa- Cry49Aa interaction, while mutations at C183 and C258 reduced the binding capacities to the larval gut cell membrane. Thus, Cysteine residues at position 91, 183, and 258 in Cry49Aa are required for full toxicity of Cry48Aa/Cry49Aa toxin.