Advances in the understanding of Blattodea evolution: Insights from phylotranscriptomics and spermathecae.

Cockroaches, an ancient and diverse group of insects on earth that originated in the Carboniferous, displays a wide array of morphology or biology diversity. The spermatheca is an organ of the insect reproductive system; the diversity of spermathecae might be the adaption to different mating and sperm storage strategies. Yet a consensus about the phylogenetic relationships among the main lineages of Blattodea and the evolution of spermatheca has not been reached until now. Here we added the transcriptome data of Anaplectidae for the first time and supplemented other family level groups (such as Blaberidae, Corydiidae) to address the pending issues. Our results showed that Blattoidea was recovered as sister to Corydioidea, which was strongly supported by molecular evidence. In Blattoidea, (Lamproblattidae + Anaplectidae) + (Cryptocercidae + Termitoidae) was strongly supported by our molecular data. In Blaberoidea, Pseudophyllodromiidae and Blaberidae were recovered to be monophyletic, while Blattellidae was found to be paraphyletic with respect to Malaccina. Ectobius sylvestris + Malaccina discoidalis formed the sister group to other Blaberoidea; Blattellidae (except Malaccina discoidalis) + Nyctiboridae was found as the sister of Blaberidae. Corydiidae was recovered to be non-monophyletic due to the embedding of Nocticola sp. Our ASR analysis of spermatheca suggested that primary spermathecae were present in the common ancestor, and it transformed at least six times during the evolutionary history of Blattodea. The evolution of spermatheca could be described as a unidirectional trend: the increased size to accommodate more sperm. Furthermore, major splits within the existing genera of cockroaches occurred in the Upper Paleogene to Neogene. Our study provides strong support for the relationship among three superfamilies and offers some new insights into the phylogeny of cockroaches. Meanwhile, this study also provides basic knowledge on the evolution of spermathecae and reproductive patterns.

Application of DNA Barcoding in the Classification of Grasshoppers (Orthoptera: Acridoidea)-A Case Study of grasshoppers from Hebei Province, China.

Grasshoppers (Orthoptera: Acridoidea) are the main pests in agriculture, animal husbandry and forestry, and some species of grasshoppers can cause serious disaster. Taxonomy is the basis of pest control. Traditional morphological identification is time-consuming and laborious. It may be due to the existence of cryptic species or the limited number of morphologists, making the identification extremely unstable. In recent years, with the development of molecular systematics, DNA barcoding technology has been applied to environment, ecology, quarantine and so on. This study focuses on testing the feasibility of DNA barcoding in the species identification for superfamily Acridoidea. Sequences of the cox1 gene were obtained from 245 individuals of 43 species of Acridoidea and one species of Tetrigoidea as outgroup from Hebei Province. Phylogenetic, genetic distance and sequence difference threshold analyses using the Maximum Likelihood (ML), Automatic Barcode Gap Discovery (ABGD) and Molecular Defined Operational Taxonomic Units (MOTU) methods, respectively, were performed for obtained sequences and the 139 additional sequences of 21 species downloaded from GenBank. The results have shown that 40, 33, and 35 species among the 48 species are consistent with the traditional morphological classification based on the phylogenetic tree, ABGD and MOTU results, respectively and the DNA barcoding technology is very efficient and helpful for identifying the species of the superfamily Acridoidea; however, the morphological approach is still playing a key role in the species identifications. It also indicates that the cox1 gene is suitable for the phylogeny of genera and species level, but it is not suitable for the phylogenetic relationship of the advanced taxa such as families.

Silencing Chitinase Genes Increases Susceptibility of Tetranychus cinnabarinus (Boisduval) to Scopoletin.

The carmine spider mite Tetranychus cinnabarinus is a major pest of crop and vegetable plants worldwide. Previous studies have shown that scopoletin is a promising acaricidal compound against Tetranychus cinnabarinus. However, the acaricidal mechanism of scopoletin remains unclear. In the present study, 12 full-length cDNAs of chitinase (CHIT) genes from Tetranychus cinnabarinus (designated TcCHITs) were cloned and characterized. Although TcCHITs were expressed throughout all life stages, their expression levels were significantly upregulated during the larval and nymphal stages. TcCHITs were downregulated 24 h after treatment with scopoletin and upregulated 24 h after treatment with diflubenzuron (DFB, a chitin synthesis inhibitor). Feeding double-stranded RNA effectively silenced TcCHIT transcription in Tetranychus cinnabarinus, thus increasing its susceptibility to scopoletin but reducing that to DFB. Meanwhile, TcCHIT silencing in larvae and adult resulted in an extremely low molting rate (7.3%) and high mortality rate (53.3%), respectively, compared with those in the control group. CHIT genes are closely related to arthropod survival, molting, and development in Tetranychus cinnabarinus, suggesting that acaricidal mechanisms of scopoletin and DFB may occur by inhibition and activation of CHIT gene expression, respectively. TcCHIT constitutes a possible target of scopoletin and DFB in Tetranychus cinnabarinus.

N,N'-di-(m-methylphenyi)-3,6-dimethyl-1,4-dihydro-1,2,4,5-tetrazine-1,4-dicarboamide (ZGDHu-1) suppresses the proliferation of PANC-1 pancreatic cancer cells via apoptosis and G2/M cell cycle arrest.

Pancreatic cancer is one of the human gastrointestinal malignancies with a high mortality and poor prognosis. Approximately eighty percent of patients are diagnosed with unresectable or metastatic disease. Thus, development of novel chemicals in the treatment of pancreatic cancer is imperative. This study aimed to investigate the anticancer effects of N,N'-di-(m-methylphenyi)-3,6-dimethyl-1,4-dihydro-1,2,4,5-tetrazine-1,4-dicarboamide (ZGDHu-1), a new tetrazine derivative, on the PANC-1 pancreatic cancer cell line and clarify the underlying molecular mechanism. Using an MTT assay, we found that ZGDHu-1 significantly suppressed the proliferation of PANC-1 cells in a time- and dose-dependent manner. Moreover, according to the morphological and flow cytometric analysis, the results indicated that ZGDHu-1 induced PANC-1 cell apoptosis and G2/M cell cycle arrest in a dose-dependent manner. In the western blot analysis, expression of the pro-apoptotic Bax gene was upregulated while the anti-apoptotic Bcl-2 gene was downregulated following treatment with ZGDHu-1. ZGDHu-1 also activated pro-caspase-3 and PARP and increased the expression of NF-κB inhibitor IκB. Furthermore, the expression levels of G2/M regulatory molecules such as cyclin B1 and cdc2 were decreased while that of Chk1 was increased. These results suggested that ZGDHu-1 suppressed the proliferation of pancreatic cancer cells, rendering it a potential therapeutic drug for the treatment of pancreatic cancer.

Removal of soluble microbial products as the precursors of disinfection by-products in drinking water supplies.

Water pollution worsens the problem of disinfection by-products (DBPs) in drinking water supply. Biodegradation of wastewater organics produces soluble microbial products (SMPs), which can be important DBP precursors. In this laboratory study, a number of enhanced water treatment methods for DBP control, including enhanced coagulation, ozonation, and activated carbon adsorption, were evaluated for their effectiveness in treating SMP-containing water for the DBP reduction purpose. The results show that enhanced coagulation with alum could remove SMPs only marginally and decrease the DBP formation potential (DBPFP) of the water by less than 20%. Although ozone could cause destruction of SMPs in water, the overall DBPFP of the water did not decrease but increased after ozonation. In contrast, adsorption by granular activated carbon could remove the SMP organics from water by more than 60% and reduce the DBPFP by more than 70%. It is apparent that enhanced coagulation and ozonation are not suitable for the removal of SMPs as DBP precursors from polluted water, although enhanced coagulation has been commonly used to reduce the DBP formation caused by natural organic matter. In comparison, activated carbon adsorption is shown as a more effective means to remove the SMP content from water and hence to control the wastewater-derived DBP problem in water supply.

Green synthesis of biocompatible carboxylic curdlan-capped gold nanoparticles and its interaction with protein.

This study demonstrates a facile, green strategy for the preparation of gold nanoparticles (AuNPs) from chloroauric acid (HAuCl4) using carboxylic curdlan (Cc) as both reducing and stabilizing agent. The as-prepared AuNPs are characterized by UV-vis spectroscopy, high resolution transmission electron microscopy, X-ray diffraction, energy dispersive X-ray spectrometry and Fourier transform infrared spectroscopy. The results indicated that the particle size of the AuNPs changes with variations in the reaction time and concentrations of Cc and HAuCl4. The spherical AuNPs are well dispersed, exhibiting high stability even after six months storage. The carboxylic groups (COO(-)) in the Cc molecules tend to adsorb and stabilize the surface of the AuNPs. The interaction between BSA and the Cc-capped AuNPs was investigated using fluorescence and circular dichroism spectroscopies. The results indicated that the BSA molecules adsorb on the surface of the AuNPs, without significant change in its helical structure even after conjugation with the AuNPs.

Protective effects of Sapindus saponins in spontaneously hypertensive rats.

OBJECTIVES: To investigate the protective effects of Sapindus saponins in spontaneously hypertensive rats, and the possible cellular and molecular mechanisms. METHODS: Thirty-two 16-week-old spontaneously hypertensive rats were randomly divided into four groups (8 in each group): model group (placebo), positive control group (27 mg/kg of Captopril Tablets), Sapindus saponins groups (27 mg/kg and 108 mg/kg, respectively). Another 8 healthy Wistar-Kyoto strain (WKY) rats were used as the normal group. The animals were treated for 8 weeks. Blood pressure of rats was determined by non-invasive blood pressure meter (BP-6). Furthermore, the contents of angiotensin II (Ang II) in plasma and myocardial tissue were determined by enzyme-linked immunosorbent assay (ELISA), the gene expression of receptor angiotensin type 1 (AT1R) in aorta was determined by quantitative realtime polymerase chain reaction (qRT-PCR). The protein expression of transforming growth factor-ß1 (TGF-ß1) and AT1R in heart was determined by immunohistochemical staining. The protein expression of p-phosphorylation of p38 mitogen-activated protein kinase (p-p38MAPK) was determined by Western blotting. The contents of interleukin (IL)-1, IL-6 and tumor necrosis factor (TNF) in serum were determined by radioimmunoassay. And the histopathological and morphological changes of aorta and heart tissue samples were assessed semi-quantitatively by hematoxylin-eosin (HE) or Masson staining. RESULTS: Thirty minutes after single or continuous treatment, systolic blood pressure (SBP) was reduced significantly in Sapindus saponins groups. And the contents of AngII, IL-1, IL-6 and TNF-α in serum, the expression of AT1R mRNA, p-p38MAPK and TGF-ß1 were significantly suppressed dose-dependently (P<0.05 or P<0.01). With the Sapindus saponins treatment, compared with those of the model group, the cardiac and aortic pathological changes were ameliorated significantly. CONCLUSIONS: Our findings suggest that Sapindus saponins might have protective effects in spontaneously hypertensive rats, the cellular and molecular mechanisms of which might be relevant to the regulation of inflammatory responses mediated by p-p38MAPK signal pathway based on activated Ang II and AT1R.

Association of HLA-DQA1 (rs9272219) with susceptibility to rheumatoid arthritis in a Han Chinese population.

HLA-DQA1 (rs9272219) has been previously reported that it is a susceptibility locus in rheumatoid arthritis (RA) of UK Caucasian population and North American; however, it has not reported in RA of Chinese population. Our study was to identify whether or not this relationship is reside between rs9272219 and RA in a Han Chinese population. 207 patients with RA and 199 control subjects were recruited. The single nucleotide polymorphism (SNP) of rs9272219 was tested in alleles and genotype frequencies and the data was analyzed by doing the statistic analysis of odds ratio (OR) and 95% confidence interval (CI) from multivariate unconditional logistic regression analyses after pairwise linkage disequilibrium (LD) was estimated. Finally, the Alleles and genotype frequencies distribution of rs9272219 locus among RA patients and control subjects were in accordance with Hardy-Weinberg equilibrium. We found significant association between rs9272219 and RA of Chinese population (OR 0.494, 95% confidence interval [95% CI] 0.354-0.688, P = 0 and OR 2.541, 95% CI 1.695-3.808, P = 0, respectively). In this study, we found that the SNP of rs9272219 in HLA-DQA1 is a potential susceptibility locus in RA of Han Chinese population; the results suggest that HLA-DQA1 may be related to the development of RA.

Characterization of soluble microbial products as precursors of disinfection byproducts in drinking water supply.

Water pollution by wastewater discharge can cause the problem of disinfection byproducts (DBPs) in drinking water supply. In this study, DBP formation characteristics of soluble microbial products (SMPs) as the main products of wastewater organic biodegradation were investigated. The results show that SMPs can act as DBP precursors in simulated wastewater biodegradation process. Under the experimental conditions, stabilized SMPs had DBPFP (DBP formation potential) yield of around 5.6 µmol mmol(-1)-DOC (dissolved organic carbon) and DBP speciation profile different from that of the conventional precursor, natural organic matter (NOM). SMPs contained polysaccharides, proteins, and humic-like substances, and the latter two groups can act as reactive DBP precursors. SMP fraction with molecular weight of <1 kDa accounted for 85% of the organic carbon and 65% of the DBP formation. As small SMP molecules are more difficult to remove by conventional water treatment processes, more efforts are needed to control wastewater-derived DBP problem in water resource management.

[Effect of Sapindus saponins on myocardial inflammation and left ventricular remodeling in spontaneously hypertensive rats].

OBJECTIVE: To investigate the effect of Sapindus saponins on myocardial inflammation and left ventricular remodeling in spontaneously hypertensive rats. METHODS: Forty 16-week-old spontaneously hypertensive rats were randomly divided into five groups, placebo as model group, captopril tablets (27 mg/kg) as positive control, low-dose Sapindus saponins (27 mg/kg), medium-dose (54 mg/ kg) and high-dose (108 mg/kg) groups. And another eight healthy Wistar-Kyoto strain (WKY) rats were used as the normal group. The animals were treated for eight weeks, and the detection indexes were as follows: (1) Calculated left ventricular mass index (LVMI); (2) Observed the morphological changes on left ventricular myocardial tissue by HE staining; (3) Observed the collagen distribution in left ventricular myocardial by Masson staining; (4) Detected the protein expression of TGF-beta1 by immunohistochemical assay. RESULTS: Sapindus saponins could effectively reverse the left ventricular hypertrophy phenomenon in SHR, lowered LVMI, inhibited the myocardial cell hypertrophy and hyperplasia of collagen fibers, and blocked the expression level of TGF-beta1 in myocardial when compared with the SHR model group, there were significant differences (P < 0.05 or P < 0.01). CONCLUSION: Sapindus saponins can reserve the left ventricular remodeling in pathological conditions, its possible mechanism may be related to the inhibition of myocardial tissue inflammation factor of TGF-beta1.

Pharmaceuticals and personal care products (PPCPs): a review on environmental contamination in China.

Pharmaceuticals and personal care products (PPCPs) which contain diverse organic groups, such as antibiotics, hormones, antimicrobial agents, synthetic musks, etc., have raised significant concerns in recently years for their persistent input and potential threat to ecological environment and human health. China is a large country with high production and consumption of PPCPs for its economic development and population growth in recent years. This may result in PPCP contamination in different environmental media of China. This review summarizes the current contamination status of different environment media, including sewage, surface water, sludge, sediments, soil, and wild animals, in China by PPCPs. The human body burden and adverse effects derived from PPCPs are also evaluated. Based on this review, it has been concluded that more contamination information of aquatic environment and wildlife as well as human body burden of PPCPs in different areas of China is urgent. Studies about their environmental behavior and control technologies need to be conducted, and acute and chronic toxicities of different PPCP groups should be investigated for assessing their potential ecological and health risks.

[Effects of sapindus saponins on inflammatory response mediated by Ang II/p38MAPK pathway and cardiac hypertrophy in spontaneously hypertensive rats].

OBJECTIVE: To investigate the effects of sapindus saponins on myocardial inflammation mediated by Ang II/ p38MAPK signal pathway and cardiac hypertrophy in spontaneously hypertensive rats. And also to explore the correlation of cardiac hypertrophy and inflammation. METHOD: Thirty-two 16-week-old spontaneously hypertensive rats (SHR) were randomly divided into four groups, one with placebo as model group, one with captopril tablets (27 mg x kg(-1)) as positive control, one with low-dose sapindus saponins (27 mg x kg(-1)), one with high-dose (108 mg x kg(-1)). And another eight healthy Wistar-Kyoto strain (WKY) rats were used as the normal group. The animals were treated for eight weeks, and the indicators detected were as follows: (1) left ventricular mass index (LVMI); (2) the content of Ang II and hs-CRP in plasma were determined by ELISA; (3) the protein expression of AT1R and VEGF were determined by immunohistochemical method; (4) the protein expression of p-p38MAPK in myocardial cells was determined by Western blot. RESULT: Sapindus saponins reduced LVMI, and blocked the expression level of Ang II, AT1R, p-p38MAPK, VEGF and hs-CRP in myocardial tissue. Vs the SHR model group, there were significant differences (P < 0.05 or P < 0.01). CONCLUSION: Our findings suggested that sapindus saponins could inhibited cardiac hypertrophy, the possible mechanisms may be related to the inhibition on inflammatory response mediated by Ang II/p38MAPK pathway.

Equity of health service utilization of urban residents: data from a western Chinese city.

BACKGROUND: Getting medical treatment is still difficult and expensive in western China. Improving the equity of basic health services is one of the tasks of the new healthcare reform in China. This study aimed to analyze the parallel and vertical equity of health service utilization of urban residents and then find its influencing factors. METHODS: In August 2011, a household survey was conducted at 18 communities of Baoji City by multi-stage stratified random sampling. Based on the survey data, we calculated a concentration index of health service utilization for different income residents and a difference index of different ages. We then investigated the influencing factors of health service utilization by employing the Logistic regression model and log-linear regression model. RESULTS: The two-week morbidity rate of sampled residents was 19.43%, the morbidity rate of chronic diseases was 21.68%, and the required hospitalization rate after medical diagnosis was 11.36%. Among out-patient service utilization, the two-week out-patient rate, number of two-week out-patients, and out-patient expense had good parallel and vertical equity, while out-patient compensation expense had poor parallel and vertical equity. The inpatient service utilization, hospitalization rate, number of inpatients, days stayed in the hospital, and inpatient expense had good parallel equity, while inpatient compensation expense had poor parallel equity. While the hospitalization rate and number of inpatients had vertical equity, the days stayed in hospital, inpatient expense, and inpatient compensation expense had vertical inequity. CONCLUSIONS: Urban residents' health was at a low level and there was not good health service utilization. There existed rather poor equity of out-patient compensation expense. The equity of inpatient service utilization was quite poor. Income difference and the type of medical insurance had great effects on the equity of health service utilization.

Biodegradation and biotransformation of wastewater organics as precursors of disinfection byproducts in water.

Laboratory experiments were carried out to investigate wastewater organics as the precursors of disinfection byproducts (DBPs) in drinking water supply. The focus was on the change in wastewater DBP precursors during biological degradation under simulated natural conditions. The wastewater and its treated secondary effluent were characterized for DBP formation potential (DBPFP) and DBP speciation profile, including trihalomethanes, haloacetic acids, chloral hydrate, and nitrogen-containing DBPs. Several model organic compounds, including humic acid, tannic acid, glucose, starch, glycine, and bovine serum albumin (BSA), were used to represent the different types of organic pollutants in wastewater discharge. The results show that the DBPFP of wastewater decreased after biodegradation, but the remaining organic matter had a greater DBPFP yield with chlorine. Different model organics displayed different changes in DBPFP during biodegradation. The DBPFP remained largely unchanged for the glycine solution, decreased greatly for the tannic acid and BSA solutions, and increased nearly 3-fold for the glucose and starch solutions after 10d of biodegradation. Meanwhile, the DBPFP yield increased from 3 for glycine to 51µg DBP mg(-1) C for its degradation residue, and from 1 for glucose and starch to 87 and 38µg DBP mg(-1) C for their organic residues, respectively. Although biodegradation may effectively remove some DBP precursors, biotransformation during the process produces new DBP precursors in the form of soluble microbial products (SMPs). The experimental results reveal that SMPs may be an important source of wastewater-derived DBP precursors in natural waters.

[Construction and characteristics of a recombinant strain apx II C- /apx I A+ of Actinobacillus pleuropneumoniae serotype 7].

An Actinobacillus pleuropneumoniae apx II C mutant was constructed by transconjugation and counterselection method. Briefly, a transconjugation plasmid pEHA1 was constructed, and transformed into donor strain Escherichia coli 32155. After mixed the donor cells with A . pleuropneumoniae acceptor cells, the mixture was cultivated for about 5 hours and plated on solid medium containing chloromycetin. Then the Cm(R) positive clones were picked and inoculated into liquid medium in the absence of any antibiotic. Cultures were pelleted, plated on sucrose plates and incubated overnight. Finally, Sucrose-resistant colonies (Suc(R)) were selected and considered as mutant. The mutant was verified by PCR, heredity stability, exotoxin secretion and sequence analysis, suggested that the construction of the mutant was sucessful. The biological characteristics of this mutant strain was further investigated. Compared with parental strain, the results indicated that the mutant hold the same growth rate in vitro and reduced virulence on mice. Altogether, this mutation system will facilitate development of live attenuated vaccines and research on functions of novel genes of A. pleuropneumoniae.