Long-term toxicity evaluation of aristolochic acid-IIIa in mice.

Aristolochic acid (AA)-IIIa is an AA analog present in Aristolochiaceae plants. To evaluate the chronic toxicity of AA-IIIa, mice were intragastrically administered with media control, 1 mg/kg AA-IIIa, and 10 mg/kg AA-IIIa, and designated as the control (CTL), AA-IIIa low dose (AA-IIIa-L), and AA-IIIa high dose (AA-IIIa-H) groups, respectively. AA-IIIa was administered three times a week, every other day, for 24 weeks (24-week time point). Thereafter, some mice were sacrificed immediately, while others were sacrificed 29 or 50 weeks after AA-IIIa withdrawal (53- or 74-week time point). Serum and organs were collected for biochemical and pathological analyses, respectively. Whole-genome sequencing was performed on the kidney, liver, and stomach tissues of AA-IIIa-treated mice for single-nucleotide polymorphism (SNP) detection. AA-IIIa-H mice died at 66 weeks, and the remaining mice showed moribund conditions at the 69 weeks. AA-IIIa induced minor kidney tubule injury, fibroblast hyperplasia, and forestomach carcinoma in mice. Bladder, intestine, liver, heart, spleen, lung, and testis tissues were not pathologically altered by AA-IIIa. In addition, AA-IIIa increased the C:G > A:T mutation in the kidney; however, no SNP mutation changes were observed in the liver and forestomach tissues of AA-IIIa-H mice at the 24-week time point compared with control mice. Therefore, we suspect that AA-IIIa is potentially mutagenic for mice after overdose and long-term administration. On the other hand, the forestomach is a unique organ in mice, but it does not exist in humans; thus, we hypothesize that the stomach toxicity induced by AA-IIIa is not a suitable reference for toxicological evaluation in humans. We recommend that Aristolochiaceae plants containing AA-IIIa should be properly supervised, and overdosing and long-term administration of drugs containing AA-IIIa should be avoided.

Recent advances and research progress on microsystems and bioeffects of terahertz neuromodulation.

Terahertz waves can interact with the nervous system of organisms under certain conditions. Compared to common optical modulation methods, terahertz waves have the advantages of low photon energy and low risk; therefore, the use of terahertz waves to regulate the nervous system is a promising new method of neuromodulation. However, most of the research has focused on the use of terahertz technology for biodetection, while relatively little research has been carried out on the biological effects of terahertz radiation on the nervous system, and there are almost no review papers on this topic. In the present article, we begin by reviewing principles and objects of research regarding the biological effects of terahertz radiation and summarizing the current state of related research from a variety of aspects, including the bioeffects of terahertz radiation on neurons in vivo and in vitro, novel regulation and detection methods with terahertz radiation devices and neural microelectrode arrays, and theoretical simulations of neural information encoding and decoding. In addition, we discuss the main problems and their possible causes and give some recommendations on possible future breakthroughs. This paper will provide insight and assistance to researchers in the fields of neuroscience, terahertz technology and biomedicine.

Zinc-Organometallic Framework Vaccine Controlled-Release Zn2+ Regulates Tumor Extracellular Matrix Degradation Potentiate Efficacy of Immunotherapy.

Tumor extracellular matrix (ECM) not only forms a physical barrier for T cells infiltration, but also regulates multiple immunosuppressive pathways, which is an important reason for immunotherapy failure. The cyclic guanosine monophosphate-adenosine monophosphate synthase-stimulator of interferon genes (cGAS-STING) pathway plays a key role in activating CD8+ T cells, maintaining CD8+ T cells stemness and enhancing the antitumor effect. Herein, a zinc-organometallic framework vaccine (ZPM@OVA-CpG) prepared by self-assembly, which achieves site-directed release of Zn2+ in dendritic cell (DC) lysosomes and tumor microenvironment under acidic conditions, is reported. The vaccine actively targets DC, significantly enhances cGAS-STING signal, promotes DC maturation and antigen cross-presentation, and induces strong activation of CD8+ T cells. Meanwhile, the vaccine reaches the tumor site, releasing Zn2+ , significantly up-regulates the activity of matrix metalloproteinase-2, degrades various collagen components of tumor ECM, effectively alleviates immune suppression, and significantly enhances the tumor infiltration and killing of CD8+ T cells. ZPM@OVA-CpG vaccine not only solves the problem of low antigen delivery efficiency and weak CD8+ T cells activation ability, but also achieves the degradation of tumor ECM via the vaccine for the first time, providing a promising therapeutic platform for the development of efficient novel tumor vaccines.

Study on the difference and correlation between the contents and toxicity of aristolochic acid analogues in Aristolochia plants.

ETHNOPHARMACOLOGICAL RELEVANCE: The nephrotoxicity and carcinogenicity induced by traditional Chinese medicines (TCMs) containing aristolochic acids (AAs) and related compound preparations have greatly limited their clinical application. While the toxicity of AA-I and AA-II is relatively clear, there are marked differences in the toxic effects of different types of aristolochic acid analogues (AAAs). Thus, the toxicity of TCMs containing AAAs cannot be evaluated based on the toxicity of a single compound. AIM OF THE STUDY: To systematically investigate the toxicity induced by Zhushalian (ZSL), Madouling (MDL) and Tianxianteng (TXT) as representative TCMs derived from Aristolochia. MATERIALS AND METHODS: AAA contents in ZSL, MDL and TXT were determined using HPLC. Subsequently, mice were treated for 2 weeks with high (H) and low (L) dosages of TCMs containing total AAA contents of 3 mg/kg and 1.5 mg/kg, respectively. Toxicity was evaluated using biochemical and pathological examination and was based on organ indices. Correlations between AAA contents and induced toxicity were analysed using multiple methods. RESULTS: Of the total AAA content, ZSL contained mainly AA-I and AA-II (>90%, of which AA-I accounted for 49.55%). AA-I accounted for 35.45% in MDL. TXT mainly contained AA-IVa (76.84%) and other AAAs accounted for <10%. Short-term toxicity tests indicated that ZSL and high-dose MDL induced obvious renal interstitial fibrosis and gastric injury, whereas TXT (high and low dosages) caused only slight toxicity. Correlation analysis suggested that AA-I might be the critical hazard factor for toxicity. CONCLUSIONS: The toxicity of TCMs containing AAAs cannot be generalised. The toxicity of TXT is relatively low compared with those of ZSL and MDL. The toxicity of Aristolochia depends mainly on the AA-I content; therefore, control of AA-I levels in TCMs and related compound preparations is required to reduce the risk of toxicity associated with the use of Aristolochia herbs in clinical settings.

Reduced Graphene Oxide and Gold Nanoparticles-Modified Electrochemical Aptasensor for Highly Sensitive Detection of Doxorubicin.

Doxorubicin (DOX) is the most clinically important antibiotic in cancer treatment, but its severe cardiotoxicity and other side effects limit its clinical use. Therefore, monitoring DOX concentrations during therapy is essential to improve efficacy and reduce adverse effects. Here, we fabricated a sensitive electrochemical aptasensor for DOX detection. The sensor used gold wire as the working electrode and was modified with reduced graphene oxide (rGO)/gold nanoparticles (AuNPs) to improve the sensitivity. An aptamer was used as the recognition element for the DOX. The 5' end of the aptamer was modified with a thiol group, and thus immobilized to the AuNPs, and the 3' end was modified with methylene blue, which acts as the electron mediator. The combination between the aptamer and DOX would produce a binding-induced conformation, which changes the electron transfer rate, yielding a current change that correlates with the concentration of DOX. The aptasensor exhibited good linearity in the DOX concentration range of 0.3 µM to 6 µM, with a detection limit of 0.1 µM. In addition, the aptasensor was used for DOX detection in real samples and results, and showed good recovery. The proposed electrochemical aptasensor will provide a sensitive, fast, simple, and reliable new platform for detecting DOX.

Long-term oral administration of Asarum heterotropoides f. mandshuricum (Maxim.) Kitag. decoction and its aristolochic acid analogs do not cause renal toxicity in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Asarum heterotropoides f. mandshuricum (Maxim.) Kitag. (AH) is widely used to treat influenza, COVID-19, allergic rhinitis, headache, toothache, rheumatoid arthritis, and peptic ulcer. However, its clinical use is controversial due to the concern of aristolochic acid nephropathy (AAN) caused by its component aristolochic acid analogs (AAs). AIM OF THE STUDY: The chronic toxicity of AH decoction and its main components AA IVa (AA-IVa) and aristolactam I (AL-I) was evaluated in mice. MATERIALS AND METHODS: AAs contents in AH were quantitated by liquid chromatography-mass spectrometry. A parallel design was employed to examine the potential chronic toxicity of AH decoction at doses equivalent to 0.5, 1.6, and 5.0 g/kg AH (approximately 10-100 times the clinical doses for humans) and its major AA components at doses equivalent to that in 5.0 g/kg AH to mice after consecutive daily oral administration for 12 and 24 weeks, and at 32 weeks after withdrawal for 8 weeks. RESULTS: AH crude herb contained 2.18 µg/g of AA-I, 48.49 µg/g of AA-IVa, and 14.0 µg/g of AL-I. AH decoction contained 5.45 µg/g of AA-IVa and 2.71 µg/g of AL-I. None of AA-II and AA-IIIa were detected in AH. After long-term administration of AH decoction and its major components AA-IVa and AL-I, mice showed no signs of illness or body weight changes. In addition, biochemical and pathohistological examinations showed that long-term administration of AH decoction and its major components AA-IVa and AL-I did not alter 1) serum levels of glutamic-pyruvic transaminase, glutamic oxalacetic transaminase, alkaline phosphatase, creatinine, and urea nitrogen, 2) renal tissue mRNA expression of kidney injury molecule 1 and neutrophil gelatinase-associated lipocalin, and 3) pathological morphology in the mouse liver, kidney, stomach, and bladder. CONCLUSIONS: AH has no obvious toxicity to mice and is relatively safe when it is used in the form of decoction. AA-IVa and AL-I, the two major AAs in AH, are not toxic to mice at the dose equivalent to that in the high dose of AH decoction. Considering the limited toxicological data on AH, we recommend that AH decoction medication should not overdose and the duration should not be too long.

Need for Distinctiveness Leads to Pathological Internet Use? The Perspective of Cognitive Behavioral Model.

Based on the cognitive behavioral model of pathological Internet use and the gender role theory, this present study investigated the association between the need for distinctiveness and pathological Internet use. Additionally, we explored a mediating role of maladaptive cognition in the association between the need for distinctiveness and pathological Internet use and tested whether the mediation model was moderated by gender. A sample of 745 Chinese university students (Mage = 19.92, SDage = 1.42) was studied and participants completed anonymous questionnaires regarding the need for distinctiveness, maladaptive cognition, and pathological Internet use. Results revealed that the need for distinctiveness was positively associated with pathological Internet use, and the association between the need for distinctiveness and pathological Internet use was mediated by maladaptive cognition. In addition, gender moderated the association between maladaptive cognition and pathological Internet use; the effect was stronger for female participants than male participants. The findings expanded our understanding of the dark side of seeking distinctiveness. Practically, the results suggest that policymakers and psychological practitioners consider gender in preventing and intervening in pathological Internet use.

Effects of filial piety belief on cyberbullying perpetration of Chinese university students.

Cyberbullying has become a worldwide phenomenon. Although the topic has drawn decent academic attention and many studies have been conducted on Chinese samples, variable interests in these studies have not captured the thinking and behavioral characteristics of Chinese people. Based on the dual filial piety model and self-determination theory, this study examined the effect of filial piety belief on cyberbullying perpetration and tested the mediation of relatedness need satisfaction. A total of 856 university students completed the questionnaires, including dual filial piety scale, relatedness need satisfaction scale and cyberbullying perpetration scale. The regression results found that reciprocal filial piety negatively predicted and authoritarian filial piety positively predicted cyberbullying perpetration. The SEM results showed that reciprocal filial piety and authoritarian filial piety impacted cyberbullying perpetration through the mediating effect of relatedness need satisfaction. Reciprocal filial piety was positively while authoritarian filial piety negatively correlated with relatedness need satisfaction and relatedness need satisfaction was negatively correlated with cyberbullying perpetration. The results provide a new position to understand the effect of family factors on cyberbullying perpetration by placing the topic within traditional Chinese family value.

We can't afford the candy! The influence of parenting by lying about money on mobile phone addiction and the roles of anxiety and socioeconomic status among Chinese adolescents.

Background: Extant research has indicated that parenting practice, such as harsh parenting, rejection, and neglect increases the risk of mobile phone addiction. However, no research to date has examined the association between parenting by lying about money and adolescent mobile phone addiction. Objective: The current study used a survey to test whether parenting by lying about money may be associated with adolescent mobile phone addiction. The mediation of anxiety and moderation of socioeconomic status were also examined. Materials and methods: We recruited 971 adolescents from five secondary schools in a city in central China. Of the participants, 448 (46.14%) were boys and 523 (53.86%) were girls (M age = 13.63, SD age = 1.01). Results: The results of mediation analysis indicated that parenting by lying about money positively predicted mobile phone addiction (B = 0.144, p < 0.01); parenting by lying about money positively predicted anxiety (B = 0.126, p < 0.01) and mobile phone addiction (B = 0.107, p < 0.01). Anxiety positively predicted mobile phone addiction (B = 0.293, p < 0.01). Moreover, the bias-corrected bootstrapping mediation test indicated that the process by which parenting by lying about money predicted mobile phone addiction through anxiety was significant (indirect effect = 0.037, SE = 0.011, 95% CI = [0.017, 0.059]). Conclusion: The current study suggests that parenting by lying about money may lead to mobile phone addiction through the mediation of anxiety. However, the effect was stronger for adolescents with higher level of socioeconomic status than their counterparts.

Ultra-performance chromatography-electrospray tandem mass spectrometry analysis of bile acid profiles in the enterohepatic circulation following geniposide and acetaminophen-induced liver injury.

Bile acids (BAs) play an important role in pre-diagnosing drug-induced liver injury (DILI). However, in clinical practice, different types of liver injury are characterized by different pathogeneses and pathological manifestations. Therefore, whether BAs can be used as biomarkers across different DILIs remains unclear. In this study, an ultra-performance chromatography-mass spectrometry (MS)/MS-based technique was developed for the simultaneous quantitative analysis of 31 BAs in the serum, liver, feces, urine, and intestinal contents of rats treated with acetaminophen (APAP) and geniposide to induce liver injury. The total extraction recovery for representative analytes ranged between 80.60% and 99.23% in the serum, urine, liver, feces, and intestinal contents. The correlation coefficients for all standard curves of the different matrices were at least 0.99. Validation of the BA analytical method including selectivity, residue, lower limit of quantification, accuracy, precision, matrix effect, and stability conformed with the biospecimen quality control standards of the Chinese Pharmacopoeia (version 2020). Serum biochemical and pathohistological analyses revealed APAP- and geniposide-induced hepatocellular and cholestatic DILI, respectively, with different effects on BA profiles in the enterohepatic circulation. Metabolomics further revealed that the trends in BA changes in the serum, feces, urine, and intestinal tissues were consistent between the geniposide- and APAP-treated groups. However, in the liver, the total BAs (TBA) concentration increased by 1.70 fold in the geniposide group but decreased by 43% in the APAP group compared with the control group. Multivariate analysis revealed differentially expressed BAs, including TCA, CA, and GCA, which are potential biomarkers for DILI, in the serum, liver, and urine following treatment with geniposide. Interestingly, the differentially expressed BAs in the APAP group were similar to those in the control group. Additionally, the magnitude of changes in the TBA in the urine (3.3 fold and 15.5 fold in the APAP and geniposide groups, respectively) was higher than that in the blood (290 fold and 640 fold in the APAP and geniposide groups, respectively). However, given the BA profiles after geniposide- and APAP-induced liver injury, BAs were found to be more suitable as biomarkers for diagnosing cholestatic liver injury. Overall, the BA assay developed in this study is rapid, simple, accurate, validated, sensitive, and suitable for analyzing the levels and distribution of BAs in various parts of the enterohepatic circulation.

Loving Your Parents and Treating Others Well: The Effect of Filial Piety on Cyberbullying Perpetration and Its Functional Mechanism Among Chinese Graduate Students.

Although a few studies have investigated the effect of family factors on cyberbullying perpetration, these studies have mainly focused on the roles of parents. Few studies have examined the roles of children. In traditional Chinese culture, the filial piety belief (FPB) refers to the viewpoint that considers the roles of children in a family context. We suggest that how children consider their roles in a family might also influence cyberbullying perpetration. Based on the dual filial piety model and social information processing model, this study examined the effect of FPB on cyberbullying perpetration and tested its mediation of hostile attribution bias. In total, 588 graduate students participated. The participants completed several questionnaires, including the FPB scale, the social information processing-attribution response questionnaire, and the cyberbullying scale. The results were as follows. First, reciprocal filial piety (RFP) was significantly negatively correlated with hostile attribution bias and cyberbullying perpetration, while authoritarian filial piety (AFP) was significantly positively correlated with hostile attribution bias and cyberbullying perpetration. Second, hostile attribution bias mediated the association between FPB (comprising AFP and RFP) and cyberbullying perpetration; RFP decreased hostile attribution bias, while AFP increased hostile attribution bias; and hostile attribution bias increased cyberbullying perpetration. The current study expands our understanding of the influential factors and formation mechanism of cyberbullying perpetration in Chinese culture.

Molecular analyses of the diversity and function of the family 1 ß-glucosidase-producing microbial community in compost.

The diversity and transcription efficiency of GH1 family ß-glucosidase genes were investigated in natural and inoculated composts using a DNA clone library and real-time qPCR. Compositional differences were observed in the functional communities between the two composting processes. Proteobacteria, Actinobacteria, Firmicutes, and Chloroflexi were the dominant phyla. Twenty representative ß-glucosidase genes were quantitatively analyzed from the DNA and RNA pools. Principal component analysis and Pearson's correlation analysis showed that cellulose degradation is correlated with the composition and succession of functional microbial communities, and this correlation was mainly observed in Proteobacteria and Actinobacteria. Compared with inoculated compost, the functional microbial communities in natural compost with a low diversity index exhibited a weak buffering capacity for function in response to environmental changes. This may explain the consistency and dysfunction of cellulose degradation and transcriptional regulation by dominant ß-glucosidase genes. Except for the ß-glucosidase genes encoding constitutive enzymes, individual ß-glucosidase genes responded to environmental changes more drastically than the group ß-glucosidase genes. The correlation results suggested that ß-glucosidase genes belonging to Micrococcales play an important role in the regulation of intracellular ß-glucosidase. These results indicated that the responses of functional microorganisms were different during both composting processes and were reflected at both the individual and group levels.

Dramaturgical perspective mediates the association between parenting by lying in childhood and adolescent depression and the protective role of parent-child attachment.

BACKGROUND: Recent findings reveal that individuals who experience parenting by lying in childhood may exhibit depression through adulthood. Questions remain regarding the cognitive mechanism underlying this association and whether depression manifests as early as adolescence. OBJECTIVE: Our current study used a survey to test the associations among parenting by lying in childhood, adolescent depression, dramaturgical perspective, and parent-child attachment. PARTICIPANTS AND SETTING: Adolescents (n = 964, Mage = 13.62, SDage = 1.04) were recruited from five secondary schools in a city in central China. A total of 446 (46.3 %) boys and 518 (53.7 %) girls participated in the survey. METHODS: We used questionnaires to measure parenting by lying, dramaturgical perspective, parent-child attachment and depression. RESULTS: A bias-corrected bootstrapping mediation test indicated that the process by which parenting by lying predicted depression through a dramaturgical perspective was significant,ab = .06, SE = .01, 95 % CI = [.027, .089]. Moderation analysis indicated that the interaction between parenting by lying and parent-child attachment was associated with depression (B = - .05, p < .05). CONCLUSIONS: Parenting by lying in childhood was found to be positively associated with adolescent depression. The association was mediated by the child's use of a dramaturgical perspective and moderated by parent-child attachment. This study highlights an intriguing but under-researched parenting practice and extends our understanding of its association with adolescent depression. In practice, strengthening parent-child attachment can weaken the association between parenting by lying and depression.

Application of smartphone in detection of thin-layer chromatography: Case of salvia miltiorrhiza.

In this work, a smartphone-based device was constructed for thin-layer chromatography (TLC) detection and semi-quantitative analysis of the components of Salvia miltiorrhiza. The key construction and shooting parameters were investigated by the relative peak area and signal-to-noise ratio. The best conditions were as follows: shooting height, 17 cm; angle between the UV lamp and TLC plate, 58°; exposure compensation, 0~0.2 EV; and shutter speed under daylight and UV 365 nm, 1/50 s and 1/5 s, respectively. These ideal conditions could be replicated by smartphones from different brands with different versions of software. With good precision, repeatability and stability, the developed device was used for the semi-quantitative analysis of salvianolic acid B, rosmarinic acid, cryptotanshinone, tanshinone I, tanshinone IIA, and miltirone in the TLC analysis of 10 batches of S. miltiorrhiza. The results were compared with those obtained by a TLC densitometric scanner and two common types of image processing software, i.e., Gelanalyzer and ImageJ. Except for salvianolic acid B in the TLC densitometric scanner, all results were not significantly different among these methods, which suggested that smartphones might be a useful tool for the quality control of traditional Chinese medicines.

Ganoderma spore powder contains little triterpenoids.

BACKGROUND: Ganoderma spore is a minuscule germ cell ejected from Ganoderma gills during its growth maturity period, it has been considered with high exploitable potential in health-care products manufacture. METHODS: After testing sporoderm-broken rate, the triterpenoids in 12 batches of broken and unbroken Ganoderma spore powder (GSP) samples were compared with Ganoderma lucidum fruiting body (GL) by high performance thin-layer chromatography (HPTLC) and further verified by liquid chromatography coupled with electrospray ionization quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS); meanwhile, the dissolution of triterpenoids after bionic extraction was also investigated by HPTLC. RESULTS: The sporoderm-broken rate of all the broken GSP samples was over 85%. The relative peak area of triterpenoids in GSP samples were lower than 50% of that in fruiting body, and the dissolution of triterpenoids in artificial gastrointestinal fluid was lower than in methanol. CONCLUSIONS: This study demonstrated that there were little triterpenoids in GSP. Triterpenoids in GSP also seldom be dissolved in artificial gastrointestinal fluid.

Decoding active components in a formulation of multiple herbs for treatment of psoriasis based on three cell lines fishing and liquid chromatography-mass spectrometry analysis.

Chinese medicine formula (CMF) usually contains multiple herbs and its components are very complicated, which therapeutic effects are derived from the integrative activity of multiple components. Therefore, the screening of bioactive compounds in the formula is necessary, though it is very difficult, not only for elucidating the therapeutic components but also for developing the quality control method. In this work, three cell lines fishing combined with LC-MS analysis was developed for screening the potential bioactive compounds in Fuzhengzhiyanghefuzhiyang decoction (FZHFZY), a CMF for the treatment of psoriasis in clinic. Nineteen potential active compounds were discovered and thirteen of them were identified as rehmannioside D, melittoside or rehmannioside A, tufulingoside, neoastilbin, astilbin, neoisoastilbin, isoastilbin, engeletin, isoengeletin, spinosin, cnidimoside A, rutaevin and limonin, respectively. The effects of identified components were also analyzed in vitro. The results showed that IL-1ß expression of RAW 264.7 cells treated with spinosin and melittoside as well as the TNF-α level of RAW 264.7 cells treated with rehmannioside D and melittoside were not significantly different compared with lipopolysaccharide (LPS)-treated cells. Though all identified potential active components had no apoptosis effect to HaCaT cells, rehmannioside D, rehmannioside A, astilbin and neoisoastilbin could significantly suppress HUVEC cells migration compared with control. These results indicated that the target cell fishing combined with LC-MS analysis is a useful tool for screening bioactive compounds from complicated CMF, and the identified active components may mainly contribute to the therapeutic effects of FZHFZY.

High-Performance Thin-Layer Chromatographic Fingerprints of Triterpenoids for Distinguishing Between Isodon lophanthoides and Isodon lophanthoides var. gerardianus.

Background: The aerial parts of Isodon lophanthoides (Buch. Ham. ex D. Don) Hara (IL) has been officially recorded as Isodonis lophanthoidis herba by many provincial quality control standards for Chinese herbal medicines in China. Recently, it has been found that one of its varieties, I. lophanthoides var. gerardianus (Benth.) Hara (ILVG) was pretended to be I. lophanthoidis herba in herbal material markets or cultivated bases. Because of the similarity on appearance, these two close-related species were difficult to be identified by morphological characters, especially when they are dried and sliced. Objective: To establish a rapid and specific method for identification of the two herbal medicines. Method: In this paper, a fingerprint of triterpenoids by HPTLC coupled with a digital profiling was established to identify IL and distinguish it from its substitute, ILVG. The specific HPTLC fingerprints constructed by determining twelve batches of IL samples and thirteen batches of ILVG samples, intuitionally reflected the difference between the two species on HPTLC image and the peak-peak rations of chemical distribution. Results: Authentication results of nine batches of commercial samples by the above established HPTLC fingerprints exhibited coincident conclusion with that by morphological means. Conclusions: The HPTLC fingerprint is proven to be simple, repeatable, specific, and suitable for rapid identification of I. lophanthoidis herba. Highlights: An efficient method for identification and distinguishing Isodon lophanthoides from its substitute, I. lophanthoides var. gerardianus, was established. HPTLC fingerprints of ursane-type triterpenoides were constructed and validated by determining IL and ILVG samples.

The structural and functional contributions of ß-glucosidase-producing microbial communities to cellulose degradation in composting.

BACKGROUND: Compost habitats sustain a vast ensemble of microbes that engender the degradation of cellulose, which is an important part of global carbon cycle. ß-Glucosidase is the rate-limiting enzyme of degradation of cellulose. Thus, analysis of regulation of ß-glucosidase gene expression in composting is beneficial to a better understanding of cellulose degradation mechanism. Genetic diversity and expression of ß-glucosidase-producing microbial communities, and relationships of cellulose degradation, metabolic products and the relative enzyme activity during natural composting and inoculated composting were evaluated. RESULTS: Compared with natural composting, adding inoculation agent effectively improved the degradation of cellulose, and maintained high level of the carboxymethyl cellulose (CMCase) and ß-glucosidase activities in thermophilic phase. Gene expression analysis showed that glycoside hydrolase family 1 (GH1) family of ß-glucosidase genes contributed more to ß-glucosidase activity in the later thermophilic phase in inoculated compost. In the cooling phase of natural compost, glycoside hydrolase family 3 (GH3) family of ß-glucosidase genes contributed more to ß-glucosidase activity. Intracellular ß-glucosidase activity played a crucial role in the regulation of ß-glucosidase gene expression, and upregulation or downregulation was also determined by extracellular concentration of glucose. At sufficiently high glucose concentrations, the functional microbial community in compost was altered, which may contribute to maintaining ß-glucosidase activity despite the high glucose content. CONCLUSION: This research provides an ecological functional map of microorganisms involved in carbon metabolism in cattle manure-rice straw composting. The performance of the functional microbial groups in the two composting treatments is different, which is related to the cellulase activity and cellulose degradation, respectively.

The distribution of active ß-glucosidase-producing microbial communities in composting.

The composting ecosystem is a suitable source for the discovery of novel microorganisms and secondary metabolites. Cellulose degradation is an important part of the global carbon cycle, and ß-glucosidases complete the final step of cellulose hydrolysis by converting cellobiose to glucose. This work analyzes the succession of ß-glucosidase-producing microbial communities that persist throughout cattle manure - rice straw composting, and evaluates their metabolic activities and community advantage during the various phases of composting. Fungal and bacterial ß-glucosidase genes belonging to glycoside hydrolase families 1 and 3 (GH1 and GH3) amplified from DNA were classified and gene abundance levels were analyzed. The major reservoirs of ß-glucosidase genes were the fungal phylum Ascomycota and the bacterial phyla Firmicutes, Actinobacteria, Proteobacteria, and Deinococcus-Thermus. This indicates that a diverse microbial community utilizes cellobiose. The succession of dominant bacteria was also detected during composting. Firmicutes was the dominant bacteria in the thermophilic phase of composting; there was a shift to Actinomycetes in the maturing stage. Proteobacteria accounted for the highest proportions during the heating and thermophilic phases of composting. By contrast, the fungal phylum Ascomycota was a minor microbial community constituent in thermophilic phase of composting. Combined with the analysis of the temperature, cellulose degradation rate and the carboxymethyl cellulase and ß-glucosidase activities showed that the bacterial GH1 family ß-glucosidase genes make greater contribution in cellulose degradation at the later thermophilic stage of composting. In summary, even GH1 bacteria families ß-glucosidase genes showing low abundance in DNA may be functionally important in the later thermophilic phase of composting. The results indicate that a complex community of bacteria and fungi expresses ß-glucosidases in compost. Several ß-glucosidase-producing bacteria and fungi identified in this study may represent potential indicators of composting in cellulose degradation.

Physiological and transcriptional responses of Nitrosomonas europaea to TiO2 and ZnO nanoparticles and their mixtures.

The short-term combined effects of two most extensively used nanoparticles (NPs) TiO2 NPs (n-TiO2) and ZnO NPs (n-ZnO) versus their individual cytotoxicities on a model ammonia-oxidizing bacterium, Nitrosomonas europaea, were investigated at both physiological and transcriptional levels. n-ZnO exerted more serious impairment effects on cell morphology, cell density, membrane integrity, and ammonia monooxygenase activity than n-TiO2. However, the co-existing n-TiO2 displayed a dose-dependent mitigation effect on n-ZnO cytotoxicity. Consistently, the n-TiO2 and n-ZnO mixture-impacted global transcriptional expression profile, obtained with the whole-genome microarray technique, was more comparable to the n-TiO2-impacted one than that impacted by n-ZnO. The expressions of numerous genes associated with heavy metal scavenging, DNA repair, and oxidative stress response were less up-regulated under the binary impacts of NP mixture than n-ZnO. Moreover, only n-ZnO alone stimulated the up-regulations of heavy metal resistance genes, which further implied the capacity of co-existing n-TiO2 to alleviate n-ZnO cytotoxicity. In addition, the damage of cell membrane structures and the suppression of cell membrane biogenesis-related gene expressions under the influence of either individual NPs or their combinations strongly suggested that the interruption of cell membranes and the associated metabolic activities would probably be one of NPs' critical cytotoxicity mechanisms.