Enantioselective effect of the chiral fungicide tebuconazole on the microbiota community and antibiotic resistance genes in the soil and earthworm gut.

Tebuconazole, consisting of two enantiomers, has a high detectable rate in the soil. The residue of tebuconazole in the soil may cause risk to microbiota community. Antibiotic resistance genes (ARGs) are considered as emerging environmental contaminants, and they can be transferred vertically and horizontally between microbiota community in the soil. Until now, the enantioselective effect of tebuconazole on the microbiota community and ARGs in the soil and earthworm gut has remained largely unknown. Tebuconazole enantiomers showed different bioconcentration behaviors in earthworms. The relative abundances of bacteria belonging to Actinobacteriota, Crenarchaeota and Chloroflexi in R-(-)-tebuconazole-treated soil were higher than those in S-(+)-tebuconazole-treated soil at same concentrations. In the earthworm gut, bacteria belonging to Proteobacteria and Bacteroidota exhibited different relative abundances between the S-(+)-tebuconazole and R-(-)-tebuconazole treatments. The numbers and abundances of ARGs in the soil treated with fungicides were higher than those in the control. In earthworm gut, the diversities of ARGs in all treatments were higher than that in the control, and the relative abundances of Aminoglycoside, Chloramphenicol, Multidrug resistance genes and mobile genetic elements (MGEs) in R-(-)-tebuconazole-treated earthworm gut were higher than those in S-(+)-tebuconazole-treated earthworm gut. Most of ARGs showed a significantly positive correlation with MGEs. Based on network analysis, many ARGs may be carried by bacteria belonging to Bacteroidota and Proteobacteria. These results provide valuable information for understanding the enantioselective effect of tebuconazole on the microbiota community and ARGs.

Molecular characterization and antimicrobial activity of NK-lysin in black scraper (Thamnaconus modestus).

NK-lysin (NKL) is a positively charged antimicrobial peptide with broad-spectrum bactericidal activities. In this study, the cDNA sequence of NKL (TmNKL) from black scraper (Thamnaconus modestus) was cloned, which encodes a predicted polypeptide of 150 amino acids that contains a surfactant protein B domain with three disulfide bonds. Phylogenetically, TmNKL was most closely related to its teleost counterpart from tiger puffer (Takifugu rubripes). Expression analysis demonstrated that TmNKL transcripts were constitutively expressed in all tested tissues, with the highest expression levels in the gills. Its expression was significantly upregulated in the gills, head kidney, and spleen after infection with Vibrio parahaemolyticus. A linear peptide (TmNKLP40L) and a disulfide-type peptide (TmNKLP40O) were further synthesized and results showed that disulfide bonds are not essential for bactericidal activities of TmNKL, and that both forms of TmNKL exhibited potent bactericidal activities against 4 gram- negative bacteria, including V. parahaemolyticus, V. alginolyticus, Edwardsiella tarda, and V. harveyi. Observed antimicrobial activities are likely due to the effects of TmNKLP40L and TmNKLP40O treatment on disrupting the integrity of both inner and outer membrane of V. parahaemolyticus, resulting in hydrolysis of bacterial genomic DNA. Damaged cell membranes and leakage of intracellular contents were further confirmed using scanning and transmission microscopy. Moreover, administration of 1.0 µg/g TmNKLP40L or TmNKLP40O significantly decreased bacterial load in tissues and thus, pronouncedly enhanced the survival of V. parahaemolyticus-infected fish. Overall, our results demonstrated that TmNKL is a potent innate effector and provides protective effects against bacterial infection.

[Nucleosides-based identification model for Fritillariae Cirrhosae Bulbus].

A high performance liquid chromatography( HPLC) method was established for the fast,and precise determination of ten nucleosides in Fritillariae Cirrhosae Bulbus and its counterfeits. Then multivariate statistical analyses,such as clustering analysis,principal component analysis( PCA),and Fisher' s linear discriminant analysis( LDA),were conducted to establish a discriminant function model for an integrated analysis. The results indicated that data acquisition time of a single sample was shortened within 16 min by the HPLC method. In the range of 5-1 000 mg·kg~(-1),the mass concentrations of all nucleosides exhibited good linear relationships with the corresponding peak areas( R2> 0. 999). The spiked recoveries were in the range of 93. 83%-108. 9% with RSDs of0. 12%-1. 3%( n = 5). The limit of quantitation( LOQ) was 0. 98-4. 13 mg·kg~(-1). As revealed by the clustering analysis,Fritillariae Cirrhosae Bulbus and the counterfeits could be discriminated into two clusters based on the content of nucleosides. Fisher's LDA could achieve this discrimination,while PCA dimension reduction failed. The accuracy of the discriminant function model established on the screened characteristic indicators reached 97. 5%. The present study proposed a new identification method of Fritillariae Cirrhosae Bulbus with one-dimensional indicators,which is simple,accurate,and reliable. It can provide a scientific basis for further optimizing the identification techniques for Fritillariae Cirrhosae Bulbus and inspiration for quality control strategy development of Chinese medicinal materials.

[Ningmitai Capsules relieve lower urinary tract symptoms in patients with benign prostatic hyperplasia : A short-term clinical observation].

OBJECTIVE: To observe the clinical effect and safety of the Chinese patent medicine Ningmitai Capsules (NMT) in relieving lower urinary tract symptoms (LUTS) in the patient with benign prostatic hyperplasia (BPH). METHODS: We randomly assigned 40 BPH patients to an experimental and a control group of equal number to receive oral administration of NMT at 4 capsules tid and terazosin hydrochloride tablets at 2 mg qd, respectively, both for 14 days. At 7 and 14 days after medication, we recorded and compared the International Prostate Symptoms Score (IPSS), maximum urinary flow rate (Qmax), quality of life (QoL) scores, results of urinalysis and blood routine examination, and indexes of hepatic and renal function. RESULTS: Both NMT and terazosin significantly improved the total IPSS score, the IPSS scores in the storage and voiding phases, increased Qmax and urine output, reduced post-void residual urine (PVR), and improved the QoL of the patients. The patients of the NMT group showed a better relief of incomplete bladder emptying, more improved QoL and fewer adverse reactions, while those treated with terazosin achieved a better attenuation of weak urine stream and PVR. CONCLUSIONS: NMT is safe and effective in relieving LUTS in BPH patients. Each of NMT and terazosin has its own advantages in attenuating urinary tract irritation and obstruction, but whether their combination may produce a better effect on LUTS and the specific mechanisms of NMT improving acute symptoms of BPH are yet to be further studied.

[Effects of sildenafil on bladder compliance and endothelin-1 in rabbit model of partial bladder outlet obstruction].

OBJECTIVE: To explore the effects of sildenafil on bladder compliance and endothelin-1 in the rabbit model of partial bladder outlet obstruction. METHODS: A total of 24 adult male New Zealand white rabbits were randomly divided into group A, group B, group C and group D (n = 6 each). The rabbit model of partial bladder outlet obstruction was established in groups C and D while groups A and B underwent a sham operation. Daily sildenafil (10 mg/kg) was dosed to groups B and C by lavage. Daily normal saline was dosed similarly to groups A and D. Bladder urodynamic examinations were conducted in each group at Week 16. Then bladder was isolated and weighed from each group. And ET-1 in bladder tissue was measured by ELISA. RESULTS: Pressure thresholds for voiding (PT) in A-D groups were (10.6 ± 2.0), (11.6 ± 2.7), (14.0 ± 4.2) and (20.4 ± 6.1) cm H2O respectively. Compared with groups A, B and C, PT in group D was significantly higher (all P < 0.01). Bladder compliance in 4 groups were (2.75 ± 0.51), (2.78 ± 0.46), (4.98 ± 2.15) and (1.22 ± 0.25) ml/cm H2O respectively. Compared with groups A, B and C, bladder compliance was significantly lower in group D (all P < 0.01). Bladder compliance in group C was higher than that in groups A and B (both P < 0.01). The weights of bladder specimens in 4 groups were (5.0 ± 0.4), (4.6 ± 0.4), (8.2 ± 1.3) and (17.9 ± 2.3) g respectively. Compared with groups A and B, the weights of groups C and D were significantly heavier (all P < 0.01). And the weight of group D was much greater than that of group C (P < 0.01). The contents of ET-1 in bladder tissue of 4 groups were (72 ± 19), (69 ± 18), (76 ± 21) and (106 ± 29) pg/g respectively. Compared with groups A, B and C, ET-1 in bladder tissue was significantly higher in group D (all P < 0.05). CONCLUSIONS: Daily sildenafil can effectively alleviate the damage of rabbit bladder compliance from partial bladder outlet obstruction and protect bladder functions. Its mechanism may be related with the down-regulation of ET-1 in bladder tissue of partial bladder outlet obstruction.

[Measuring fatty acid concentration in maize grain by near-infrared reflectance spectroscopy].

The fatty acid concentrations in maize grain were analyzed with a set of 294 samples including normal inbred lines, high-oil inbred lines and high-oil recombinant inbred lines (RIL). The method of partial least squares (PLS) regression with internal cross validation was employed to develop the measuring models of near-infrared reflectance spectroscopy (NIRS) for concentrations of four major fatty acids, palmitic, stearic, oleic and linoleic acids, as well as oil concentration in maize grain. The NIRS models were accurate for oleic acid, linoleic acid and oil concentrations. The determination coefficients of these models in cross validation were 0.89, 0.88 and 0.91, respectively; the determination coefficients in external validation were 0.86, 0.84 and 0.92, respectively; and the ratio of standard deviation (SD) to root mean square error of validation (RMSEV) in both calibration and external validation sets (RSC(P)) was higher than 2.5. But the models for palmitic and stearic acid concentrations were not accurate enough with determination coefficients in cross validation and external validation lower than 0.80, and RSC(P) lower than 2.5. Further practical validation showed that the predicted results by using NIRS models for oleic acid, linoleic acid and oil concentrations were accurate and reliable, which will be a useful approach to the measurement of a large number of breeding samples during genetic improvement of oil quality and quantity in maize.

[Genetic diversity of D-genome revealed by SSR markers in synthesized hexaploid wheat introduced from CIMMYT].

Simple sequence repeat (SSR) molecular marker was used to measure the genetic diversity of D-genome in 26 synthesized hexaploid wheat (AABBDD) introduced from CIMMYT. Twenty-three D-genome specific SSR primers were selected for PCR amplification, among which 22 primers can detect polymorphism. A total of 92 alleles were identified at 23 loci using the above SSR primers, with an average of 4 alleles per locus. The 92 alleles were used to calculate Nei's similarity index (GS) and the genetic distance (GD). It was also found that the mean genetic distance between 26 synthesized hexaploid wheat was 0.4955, which was obviously high. From the above results, it can be indicated that the genetic variation of D-genome in synthesized hexaploid wheat was abundant and could be used to improve the genetic diversity in wheat breeding. Interestingly, synthesized hexaploid wheat 17 and 18 shared the same D-genome donor, but three of 23 detected SSR loci were polymorphic between the two materials. Therefore, during the period of allopolyploidization, there was genetic differentiation in repeat region of donor genome.