Objective:Based on the principle that the aggregation-induced emission (AIE) fluorescent probe 6PD-DPAN could bind and aggregate with bacteria, and the fluorescence intensity could reflect the quantity of bacteria, a new method for rapid, convenient, and accurate bacterial drug sensitivity testing was established, which provided a basis for rapid and accurate clinical drug use.Methods:This was a methodological evaluation study. A total of 107 clinical isolates were collected from Houjie Hospital of Dongguan City from January to December 2022, among which 46 isolates were used for the establishment of the new method, and 61 isolates were used for methodological validation. The minimum inhibitory concentration (MIC) determined by broth microdilution method was used as the gold standard, and three antibacterial drugs, gentamicin, levofloxacin, and cefotaxime, were used as experimental drugs. The AIE plate was incubated for 4 hours, and the fluorescence intensity was measured every half an hour to draw a fluorescence change curve. The MIC results were compared with the CLSI breakpoints to determine the bacteria as sensitive, intermediate, or resistant. To simplify the detection process, the ratio of fluorescence intensity at 4 hours(R) was calculated, and the ROC curve was used to analyze the efficacy of R in determining bacterial growth and establish its cutoff value. The new method was used to determine the MIC of 61 clinical isolates, with broth microdilution method as the gold standard. The basic consistency, categorical consistency, very major errors, and major errors of the new method were analyzed, and the consistency between the two methods was determined by the Kappa test.Results:ROC curve analysis of the R after 4 hours of culture: The cut-off value was 3.0, with both sensitivity and specificity for determining bacterial growth being 100%. The median (interquartile) R for bacterial growth inhibition was 11.1 (8.6, 14.4); the median R-value for bacterial growth was 1.1 (1.0, 1.2). Compared to the gold standard, the newly established method showed 100% (61/61) essential agreement in detecting MICs of 61 clinical isolates, with a categorical agreement of 96.7% (59/61). There were no very major or major errors, and the Kappa value was 0.94, indicating good consistency between the newly established method and the microbroth dilution method.Conclusions:This study successfully established a new method for bacterial drug sensitivity testing based on AIE technology, which could obtain satisfactory results within 5 hours, providing a basis for early precision drug treatment in clinical practice.

Circular RNAs (circRNAs) are endogenous non-coding RNAs with covalently closed structures, which have important functions in plants. However, their biogenesis, degradation, and function upon treatment with gibberellins (GAs) and auxins (1-naphthaleneacetic acid, NAA) remain unknown. Here, we systematically identified and characterized the expression patterns, evolutionary conservation, genomic features, and internal structures of circRNAs using RNase R-treated libraries from moso bamboo (Phyllostachys edulis) seedlings. Moreover, we investigated the biogenesis of circRNAs dependent on both cis- and trans-regulation. We explored the function of circRNAs, including their roles in regulating microRNA (miRNA)-related genes and modulating the alternative splicing of their linear counterparts. Importantly, we developed a customized degradome sequencing approach to detect miRNA-mediated cleavage of circRNAs. Finally, we presented a comprehensive view of the participation of circRNAs in the regulation of hormone metabolism upon treatment of bamboo seedlings with GA and NAA. Collectively, our study provides insights into the biogenesis, function, and miRNA-mediated degradation of circRNAs in moso bamboo.
RNA, Circular/metabolism* ; Multiomics ; Poaceae/metabolism* ; Seedlings/genetics* ; Hormones/metabolism* ; MicroRNAs/metabolism* ; Gene Expression Regulation, Plant

Objective: This study aimed to investigate the influence of monobutyl phthalate (MBP) on the expressions of epithelial-mesenchymal transition (EMT)-related proteins, migration and invasion of mouse Leydig tumor cells (MLTC-1) cells. Methods: After exposed to different doses of MBP (0、10^-7、10^-6, 10^-5, 10^-4, 10^-3 mol/L) for 24 h or 48 h, cell viability was determined by 3-(4 5-dimethyl-2-thiazolyl)-2 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Expressions of vimentin, E-cadherin, N-cadherin and Snail proteins related to EMT were detected by Western blot. The ability of migration and invasion of MLTC-1 were assessed by wound healing assay and Transwell Boyden chamber assay, respectively. Results: Relative expressions of vimentin, Snail and N-cadherin proteins were promoted ((1.56±0.07) vs (1.78±0.08), (1.22±0.06) vs (1.44±0.07), (1.33±0.11) vs (2.19±0.06), all P values were<0.001) and E-cadherin (0.66±0.09) vs (0.47±0.06), P<0.001,protein was inhibited after the cells stimulated with MBP (0, 10^-7 and 10^-6 mol/L). The capability of wound closure of MLTC-1 cells were (6.64±2.07)%, (15.61±2.83)%, (39.91±0.33)%, respectively and the invading/migrating cells were (32.67±3.51), (57.67±2.52), (82.67±6.51), respectively, which were obviously increased under MBP treatments (0, 10^-7 and 10^-6 mol/L) (all P values were <0.001). Conclusion Monobutyl phthalate affected the expressions of EMT-related proteins and enhanced the migration and invasion of MLTC-1 cells.

Objective To study the effects of experiment-related factors on hematological parameters in SD rats, analyze the data difference and causes, understand the effects of anesthetics and stress responses on the physiological aspects of animals, and to provide a reference for the standardization of animal welfare and compound toxicity testing methods.Methods According to gender (A), fasting time (B), anesthesia (C) and blood collection mode (D), SPF SD rats were divided into 24 groups.Blood samples were collected from each group.Then, red blood cell count, hemoglobin levels, white blood cell count and classification indicators were measured.Results The primary and secondary order of the factors affecting the white blood cell count was D > C > A > B, and the levels of white blood cell count of each factor were male rats > female rats, and venous blood > arterial blood, chloral hydrate > pentobarbital sodium > no anesthesia.The primary and secondary order of the factors affecting the white blood cell classification was C > D=A=B, and factors affecting the levels of white blood cell classification were chloral hydrate > pentobarbital sodium > no anesthesia.The primary and secondary order of the effects of the factors on the red blood cell count and hemoglobin level was C > D=A=B, and the levels of red blood cell count and hemoglobin level were pentobarbital sodium > chloral hydrate> no anesthesia.There was no significant difference in the blood indexes between the different fasting time groups.Conclusions There is no effect of fasting on hematological parameters, but there are differences in the blood parameters between arteries and veins.The effect of chloral hydrate anesthesia on the count and classification of white blood cells is greater than that of pentobarbital sodium.The effect of chloral hydrate anesthesia on the red blood cell count and hemoglobin level is greater than that of pentobarbital sodium.The two kinds of anesthesia methods have their own advantages and disadvantages.

OBJECTIVETo evaluate the effects of di-butyl phthalate(DBP) on the sperm motility and oxidative stress in rats.

METHODSHealthy 6-week-old male Sprague Dawlay rats were randomly divided into 4 groups with 8 in each group. DBP dissolved in peanut oil was administered by gavage at dosage of 0, 250, 500, 1,000 mg/(kg.d). After 4-week DBP exposure, the animals were killed and the organs were selected and weighed. The sperm VCL, VSL, VAP, BCF, ALH, LIN, MAD and STR in the cauda epididymis were assessed by computer assisted sperm analysis (CASA), and the activities of superoxide dismutase (SOD) and the contents of malondialdehyde (MDA) in serum and testis homogenate were measured simultaneously. The increase of body weight per day and the organ body weight ratio changes of the liver, testes and epididymides were also observed.

RESULTSThe liver organ body weight ratios of the treated groups were higher than those of the control (P < 0.01), while the testis organ body weight ratios were lower at dosage of 1,000 mg/(kg.d) DBP. Compared with the control group, the parameters of rat sperm VCL and ALH declined significantly at dosage of 1,000 mg/(kg.d) DBP. In addition, DBP showed inhibiting effect on SOD activities in the testis, and it was significant in the highest exposure group compared with the control (P < 0.05). However, there were no differences in serum SOD activities between the treated groups and the control.

CONCLUSIONDBP exposure may affect the sperm motility and the anti-oxidative systems. The testis is a vital target organ influenced by DBP.

Animals ; Dibutyl Phthalate ; toxicity ; Dose-Response Relationship, Drug ; Male ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; drug effects

OBJECTIVETo explore the effect of dibutyl phthalate (DBP) on the biochemical enzymes and lipid peroxidation in rats.

METHODSHealthy 6-week old male Sprague-Dawley rats were randomly divided into 4 groups with 16 in each. DBP dissolved in peanut oil was administered by gavage at dosages of 0, 250, 500 and 1000 mg/(kg x d). After 2- and 4-week DBP exposure, 8 rats in each group were killed, with certain organs selected and weighed. The activities of biochemical enzymes and glutathione peroxidase (GSH-Px) and the levels of glutathione (GSH) in the serum and testis homogenate were determined respectively.

RESULTSDBP induced a rise in the liver organ body weight ratio, but a fall in the testis organ body weight ratio, and it was significant in the highest exposure group compared with the control after either 2-week or 4-week treatment (P < 0.01). After 2-week DBP exposure, GSHPx activities in the serum and GSH levels in the testis homogenate showed a decreasing tendency, but GSHPx activities increased markedly in the testis homogenate (P < 0.05). After 4-week DBP exposure, while alkaline phosphatase (ALP) activities in the serum revealed an increasing tendency, sorbitol dehydrogenase (SDH) activities were inhibited significantly in both the serum and the testis homogenate at the dosage of 1000 mg/(kg x d) compared with the control group (P < 0.01). Furthermore, GSH contents in the serum were also affected at this dose (P < 0.05).

CONCLUSIONThe results indicate that DBP administration strongly affects the liver and the testis organ body weight ratios. Lipid peroxidation is one possible toxic mechanism caused by DBP. SDH may be one of the most sensitive toxic indices when exposed to DBP.

Alkaline Phosphatase ; metabolism ; Animals ; Dibutyl Phthalate ; toxicity ; Dose-Response Relationship, Drug ; Glutathione ; metabolism ; L-Iditol 2-Dehydrogenase ; metabolism ; Lipid Peroxidation ; drug effects ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Testis ; drug effects ; metabolism