The impact of transgenic crops on non-target organisms is a key aspect of environmental safety assessment to transgenic crops. In the present study, we fed two snail species, Bradybaena (Acusta) ...ravida (B. ravida) and Bradybaena similaris (Ferussac)(B. similaris), with the leaves of transgenic Bt cotton Zhong 30 (Z30) and control cotton, its parent line zhong 16 (Z16), to assess the environmental safety of Bt cotton to common non-target organisms in the field. Survival, body weight, shell diameter, helix number, reproduction rate, superoxide dismutase (SOD) activity and Bt protein concentration in snails were monitored in 15 days and 180 days experiments. We also monitored the population dynamics of B. ravida and B. similaris in Z30 and Z16 cotton fields for two successive years. Compared to the snails fed on the control cotton Z16, there was no significant difference in survival, growth, reproduction, and SOD activity on Bt cotton Z30. Bt protein concentrations were significantly between different treatments, and Bt protein residues were only detected in the feces of the Z30 treatment. According to the field data, the number of B. ravida and B. similaris fluctuated considerably across seasons over the entire cotton-growing season; however, there were no significant differences between the Bt and control cotton fields at similar time. As the results showed, in our experiments, Bt cotton Z30 had no adverse effects on the two snail species, both in the laboratory and in the fields.
•The effects of Bt cotton Z30 on two land snail species (B. ravida and B. similaris) were studied for the first time.•Our research includinged short-term and long-term toxicological experiments in lab and field investigation.•We set up a good platform was established to evaluate the effect of transgenic plants on land snails in the future.•We found that the helix number is more stable and reliable than body weight and diameter, which was rarely used before.
AIM： To investigate the changes of methylation state and expression of RASSF1A gene in human gastric cancer cell lines SGC7901 and BGC823 which were treated in vitro with demethlylating agent ...5-Aza-CdR in combination with histone deacetylase inhibitor NaB. METHODS： After SGC7901 and BGC823 cells were treated with 5-Aza-CdR and/or NaB, the methylation state of RASSFIA gene was detected by methylationspecific PCR, and the changes in expression of mRNA and protein level of RASSFIA gene were observed by RT-PCR and Western-blotting before and after drug treatment. RESULTS： Hypermethylation was detected in the promoter region of RASSF1A gene in both SGC7901 and BGC823 cells, and there was no expression of this gene at both mRNA and protein level. After treatment with 5-Aza-CdR, demethylation occurred in the promoter region of RASSFIA gene, which subsequently induced re-expression of this gene. The treatment with NaB alone showed no effect on the methylation state and expression of RASSFIA gene. The combined treatment of 5-Aza-CdR and NaB induced complete demethylation of RASSFIA gene, leading to a significantly higher reexpression of the mRNA and protein of RASSFIA than those treated with 5-Aza-CdR alone （P 〈 0.05）. CONCLUSION： Hypermethylation in the promoter region is related to inactivation of RASSFIA gene in human gastric cancer cell lines SGC7901 and BGC823, while demethlylating agent 5-Aza-CdR can reverse the methylation state of RASSF1A gene and induce itsre-expression. Histone deacetylase inhibitor NaB had a synergistic effect with 5-Aza-CdR in both demethylation and gene transcriptional regulation.
To investigate the effects of sodium butyrate (NaB), a histone deacetylase inhibitor (HDACI), on the proliferation of human gastric cancer cells and the expression of p16, an important ...negative-regulatory factor of the cell cycle G1.
Human gastric cancer cells of the lines SGC7901 and BGC823 were cultured in RPMI1640 medium and treated with NaB of different concentrations: 5 x10(-4), 1 x 10(-3), 3 x 10(-3), and 5 x 10(-3) mol/L for 24 h or 72 h. MTT assay, flow cytometry, and annexin V-fluorescein isothiocyanate staining were performed to analyze the cell proliferation activity, cell cycle, and apoptotic rate. RT- PCR, Western blotting, and methylation-specific PCR (MSP) were used to detect the e mRNA and protein expression and methylation state of p16 gene respectively.
Exposure to different concentrations of NaB inhibited the growth of the SGC7901 and BGC823 cells. Treated with NaB of the concentrations of 1 x 10(-3), 3 x 10(-3), and 5 x 10(-3) mol/L respectively for 72 h, the numbers of the SGC7901 and BGC
A metagenomic library consisting of 3,024 bacterial artificial chromosome clones was prepared in Escherichia coli DH10B with high-molecular-weight DNA extracted from red soil in South China. A novel ...cellulase gene with an open reading frame of 1,332 bp, cel5G, encoding an endo-β-1,4-glucanase was cloned using an activity-based screen. The deduced enzyme, Cel5G, belongs to the glycosyl hydrolase family 5 and shares <39% identity with endoglucanases in the GenBank database. cel5G was expressed in E. coli BL21, and the recombinant enzyme Cel5G was purified to homogeneity for enzymatic analysis. Cel5G hydrolyzed a wide range of β-1,4-, β-1,3/β-1,4-, or β-1,3/β-1,6-linked polysaccharides, amorphous cellulose, filter paper, and microcrystalline cellulose. Its highest activity was in 50 mM citrate buffer, pH 4.8, at 50°C. Cel5G is stable over a wide range of pH values (from 2.0 to 10.6) and is thermally stable under 60°C. It is highly tolerant and active in high salt concentrations and is stable in the presence of pepsin and pancreatin. The K m and V max values of Cel5G for carboxymethyl cellulose are 19.92 mg/ml and 1,941 μmol min⁻¹ mg⁻¹, respectively. These characteristics indicate that Cel5G has potential for industrial use.
Paratypes: Three adult males (KIZ 035741, KIZ 035742, and KIZ 035776) and one adult female (KIZ 035740) collected by S.B. Hou and P.F. Wei on 19 June 2019 from same locality as holotype; three adult ...males (KIZ 028321, KIZ 028323, and KIZ 028324) and three adult females (KIZ 028318, KIZ 028325, and KIZ 028326) collected by K. Wang and J.L. Ren on 6 July 2016 from Gongnong Village, Pantiange Town, Weixi County, Yunnan Province, China; one adult male (KIZ 044335) collected by K. Wang from Shigu Township, Lijiang City; single adult male (CIB II0447) from Weixi, Yunnan (collector and collecting information unknown; CIB: Diagnosis: Hebius weixiensis sp. nov. can be distinguished from its recognized congeners by a combination of the following characters: (1) nasal divided, each half in contact with each other; (2) loreal pentagonal, entering orbit or not; (3) supralabials eight; (4) preocular one or two; (5) postoculars three; (6) infralabials eight to ten; (7) dorsal scales 19-19-17 rows; (8) dorsal scale reduction from 19 to 17 rows at position of 96th to 101st ventrals; (9) ventrals 171-182; (10) subcaudals 74-88, paired; (11) anal divided; (12) no dorsal stripes or some dorsal stripes on posterior body; (13) no ventrolateral stripe; and (14) ventral body pale yellow. Body moderately stout, total length 567 mm (SVL 425 mm, TaL 142 mm); tail slender, 25.0% total length; head length moderate (HL 16.7 mm, 3.9% of SVL), oval in shape, distinct from neck; eyes large, pupil suborbicular; rostral semicircular, invisible from above; nasal divided, lateral nostril piercing medially; internasals paired, trapezoidal in shape, narrower end facing; prefrontals paired, larger than internasals; frontal hexagonal, spear like, longer than width, tip facing posteriorly and inlayed by both parietals; parietal paired, large, contacting five small nuchal scales; loreal pentagonal, slender, entering orbit; preocular single; supraocular single; postoculars three; temporals 2+1+3; supralabials eight, fourth and fifth entering orbit, seventh largest; mental triangle, inlayed completely by first pair of infralabials; infralabials nine; two pairs of chin shields; dorsal lanceolate, imbricated, 19-19-17 rows, distinctively keeled except outer most two rows; dorsal scale rows reduced from 19 to 17 between 99th and 101st ventrals; ventrals 175; subcaudals paired, 87 excluding tip; cloacal plate divided. Hebius septemlineatus comb. nov. can be distinguished from its recognized congeners by a combination of the following characters: (1) TaL/ToL 23.5%-25.6%; (2) internasals narrowed anteriorly; (3) nasal divided, each half in contact with each other; (4) nostrils lateral; (5) loreal almost quadrilateral, higher than width, separated from orbit; (6) supralabials usually eight; (7) preocular one or two; (8) postoculars usually three; (9) mental triangles; (10) infralabials nine or ten, first pair in contact with each other, first-fifth infralabials in contact with anterior chin shields; (11) dorsal scales 19-19-17 rows; (12) dorsal scale rows reduced from 19 to 17 at position of third and fourth dorsal scale rows and from 97th to 101st ventrals; (13) ventrals 164-175; (14) subcaudals 80-96, paired; (15) anal divided; (16) four dorsal stripes on each side, not including ventrolateral stripe; and (17) ventral pale yellow.
To investigate the effect of demethylating agent 5-Aza-CdR (5-aza-2'- deoxycytidine) on demethylation and transcription-regulating of RASSF1A gene in gastric cancer cell SGC7901 in vitro, as well as ...on the growth inhibition of cells.
After SGC7901 cells were treated with 5-Aza-CdR, MTT assay, flow cytometry, and Annexin V-FITC staining were performed to analyze the cell proliferation, cell cycle and apoptotic rate respectively. Methylation- specific PCP (MSP), RT-PCR and Western blotting were used to detect methylation state, expression of mRNA and protein of RASSF1A gene.
After SGC7901 cells were treated with different concentrations of 5-Aza-CdR, the cell growth was inhibited(P<0.05), the cell cycle was blocked at G(1) phase, and the apoptotic rate increased significantly(P<0.05). Hypermethylation was detected in the promoter region of RASSF1A gene in SGC7901 cells, and no expression of RASSF1A mRNA and protein was found. After treated with 5-Aza-CdR, demethylation occurred in RASSF1A gene,which subsequentl
Divergence of gene expression and alternative splicing is a crucial driving force in the evolution of species; to date, however the molecular mechanism remains unclear. Hybrids of closely related ...species provide a suitable model to analyze allele-specific expression (ASE) and allele-specific alternative splicing (ASS). Analysis of ASE and ASS can uncover the differences in
-regulatory elements between closely related species, while eliminating interference of
regulatory elements. Here, we provide a detailed characterization of ASE and ASS from 19 and 10 transcriptome datasets across five tissues from reciprocal-cross hybrids of horse×donkey (mule/hinny) and cattle×yak (dzo), respectively. Results showed that 4.8%-8.7% and 10.8%-16.7% of genes exhibited ASE and ASS, respectively. Notably, lncRNAs and pseudogenes were more likely to show ASE than protein-coding genes. In addition, genes showing ASE and ASS in mule/hinny were found to be involved in the regulation of muscle strength, whereas those of dzo were involved in high-altitude adaptation. In conclusion, our study demonstrated that exploration of genes showing ASE and ASS in hybrids of closely related species is feasible for species evolution research.
This meta-analysis was performed to evaluate the relationships between genetic polymorphisms in the TCF7L2 gene and polycystic ovary syndrome (PCOS) risk.
The PubMed, Centralised Information Service ...for Complementary Medicine (CISCOM), Cumulative Index to Nursing and Allied Health Literature (CINAHL), Web of Science, Google Scholar, EBSCO, Cochrane Library, and Common Biorepository Model (CBM) databases were searched for relevant articles published before November 1st, 2013, without language restrictions. Meta-analysis was conducted using the STATA 12.0 software. The relationships were evaluated by calculating the pooled odds ratios (ORs) and their 95% confidence intervals (CIs). Seven case-control studies with a total 2458 PCOS patients and 5109 healthy subjects' met our inclusion criteria for qualitative data analysis. Two common polymorphisms (rs7903146 C→T and rs12255372 G→T) in the TCF7L2 gene were assessed.
The results of our meta-analysis suggested that TCF7L2 genetic polymorphisms might be strongly correlated with an increased risk of PCOS (allele model, OR=1.33, 95% CI=1.15-1.54, P<0.001; dominant model, OR=1.40, 95% CI=1.12-1.75, P=0.003), especially for the rs7903146 C→T polymorphism. A subgroup analysis was done to investigate the effect of ethnicity on an individual's risk of PCOS. Our results revealed positive significant correlations between TCF7L2 genetic polymorphisms and an increased risk of PCOS among Caucasians (allele model, OR=1.26, 95% CI=1.08-1.47, P=0.004; dominant model, OR=1.33, 95% CI=1.00-1.76, P=0.046) and Asians (allele model, OR=2.02, 95% CI=1.42-2.89, P<0.001; dominant model, OR=2.02, 95% CI=1.40-2.92, P<0.001), but not among Africans (all P<0.05).
Our findings provide convincing evidence that TCF7L2 genetic polymorphisms may contribute to susceptibility to PCOS, especially for the rs7903146 C→T polymorphism among Caucasians and Asians.
AIM:To assess whether differential expression of caspase-3 in paired metastatic lymph nodes(LNs)is prognostic of survival in patients with resectable esophageal squamous cell ...carcinoma(ESCC).METHODS:Capases-3 expression was evaluated immunohistochemically in 122 pairs of primary ESCCs and regional metastatic LNs assembled on tissue microarrays.The impact of caspase-3 expression on survival outcomes was analyzed by the Kaplan-Meier method and Cox proportional hazards regression model.RESULTS:The level of caspase-3 expression was significantly higher in LN metastases than in primary tumors(P<0.001).Caspase-3 expression in the primary tumors was associated with longer median survival(23 mo vs21 mo,P=0.033),whereas higher expression in paired metastatic LNs was associated with shorter median survival(20 mo vs 22 mo,P=0.043).Multivariate analysis showed that both were independent prognostic factors.CONCLUSION:Caspase-3 expression in metastatic LNs may be a potential independent predictor of poorer overall survival in patients with resected ESCC and LN metastasis.Protein expression in metastatic tumors may be a biomarker prognostic of survival.
Phospholipase Dα（PLDα） is involved in plant response to salt stress,but the mechanisms remain unclear. We investigated rice PLDα（OsPLDa） localization and its effect on tonoplast（TP） and plasma ...membrane （PM） H^＋ -ATPase activity and transcription in response to NaCI.When rice suspension-cultured cells were treated with 100 mM NaCI,PLDa activity in cell extracts showed a transient activation with a threefold increase at 1 h.The amount of OsPLDαprotein decreased slightly in the cytosolic fractions,whereas it increased significantly in the TP after NaCI treatment.OsPLDα1 knockdown cells were developed using RNA interference（RNAi） methods.The increase in TP and PM H -ATPase activity induced by NaCI was significantly inhibited in OsPLDα1-RNAi cells.Knockdown of OsPLDα1 prevented the NaCI-induced increase in the transcript level of OsVHA-A（encodes TP H^＋-ATPase） and OSA2（encodes PM H^＋-ATPase）, as well as OsNHX1（encodes TP Na^＋ /H^＋ antiporter）.The cells died more in OsPLDα1-RNAi mutant than in wild type when they were treated with NaCI.These results suggest that OsPLDαis involved in salt tolerance in rice through the mediation of H^＋ -ATPase activity and transcription.
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