[Optimization ofPig-aGene Mutation in Rats and Assessment of Time-dependent and Dose-response Relationship of N-ethyl-N-nitrosourea].

OBJECTIVES: To optimize the method of Pig-a mutation assay, and to explore the time-dependent and dose-response relationship of N-ethyl-N-nitrosourea (ENU). METHODS: Thirty rats were randomly assigned to 5 groups: treated with PBS (control group)or different doses of ENU (10, 20, 40 and 80 mg/kg) for 3 d by oral gavage. Blood samples were collected at 0 d, 15 d, 30 d, 45 d, 60 d, 75 d and 90 d. After enrichment, erythrocytes were incubated with Anti-CD59-APC and SYTO 13 nucleic acid dye solution. Mutant phenotype erythrocytes (RBCCD59-) and mutant phenotype reticulocytes (RETCD59-) were measured by flow cytometry to analyze mutant frequencies, and the RET percentage was determined as well. RESULTS: The RBCCD59- mutation frequency in 4 ENU groups were significantly increased in a dose- and time-dependent manner. The RETCD59- mutation frequency increased to a stable high level with a slight fluctuation, and decreased at 45 d , with the peak values observed at 30 d. The RETCD59- mutation frequency showed a dose-dependent trend in 4 ENU groups. The RET percentage in all 5 groups declined at 30 d, to a stable low level thereafter, but the trends showed no significant differences by time or group. CONCLUSIONS: The optimized in vivo Pig-a mutation assay could detecte the mutagen, such as ENU, induces mutation in RBC in a time- and dose-dependent manner.

Improvement on the extraction method of RNA in mites and its quality test.

To solve the long-existing difficult problems in extracting RNA and constructing a complementary DNA (cDNA) library for trace mites, we conducted a further comparative experiment among three RNA extraction methods (TRIzol method, Omega method, and Azanno method) based on our previous attempts at the construction of cDNA library of mites, with Psoroptes cuniculi still used as the experimental subject. By subsequently decreasing the number of mites, the least number of mites needed for RNA extraction of each method were found by criteria of completeness, concentration, and purity of the extracted RNA. Specific primers were designed according to the allergen Pso c1, Pso c2, and Actin gene sequences of Psoroptes to test the reliability of cDNA library. The results showed that Azanno method needed only 10 mites with sensitivity 204 times higher than previously used TRIzol method and 20 times higher than Omega method; clear RNA band was detected by agarose gel electrophoresis; and ultraviolet spectrophotometer determination showed that RNA concentration, 260/280, and 260/230 were in the range of 102 to 166 ng/µl, 1.83 to 1.99, and 1.49 to 1.72, respectively. Finally, specific primers detection showed that the amplified sequences had 98.33, 98.19, and 99.52% identities with those of P. cuniculi or Psoroptes ovis in GenBank, respectively, indicating that the cDNA library constructed using 10 mites was successful and it could meet the requirements for molecular biology research. Therefore, we concluded that Azanno method was more effective than TRIzol method and Omega method in RNA extraction and cDNA library construction of trace mites.

The cardioprotective effect of Shengmai Zhenwu decoction on patients with chronic heart failure based on the levels of soluble interleukin 1 receptor-like 1.

Objective: The purpose of this study was to evaluate the protective effect of Shengmai Zhenwu decoction on patients with chronic heart failure (CHF) based on the levels of soluble interleukin 1 receptor-like 1 (ST2). Methods: We included a total of 80 outpatients and inpatients with CHF who were undergoing treatment at the Shanghai Municipal Hospital of Traditional Chinese Medicine between March 2020 and March 2022. We randomly divided them into the observation group (n = 40) and the control group (n = 40). Patients in the control group received treatments as per conventional Western medicine, while those in the observation group were treated with the Shengmai Zhenwu decoction in conjunction with Western medicine for eight consecutive weeks. We then compared the pre- and post-treatment levels of ST2 and N-terminal pro-brain natriuretic peptide (NT-proBNP) of the patients in the two groups. Results: There were no significant differences in the pre-treatment levels of ST2 and NT-proBNP indexes between the two groups (P > 0.05), while the post-treatment comparison between the two groups in terms of ST2 and NT-proBNP levels suggested that the effect in the observation group was better, with statistical significance (P < 0.05). Conclusion: Shengmai Zhenwu decoction was beneficial in patients with CHF, suggesting that it could be a promising and effective method for the treatment of CHF.

Carbonyl-bis-(triphenyl-phosphane-κP)(η(2)-1-vinyl-pyrrolidin-2-one-κO)ruthenium(0).

The 1-vinyl-pyrrolidin-2-one ligand in the title compound, [Ru(C(6)H(9)NO)(C(18)H(15)P)(2)(CO)], coordinates to the Ru(0) atom with the olefin double bond and the ketone O atom. The Ru(0) atom adopts a distorted trigonal-bipyramidal coordination geometry, with the C O ligand and the ketone O atom occupying the axial positions. The two triphenyl-phosphane ligands are cis to each other. The olefinic C=C bond is almost coplanar with the Ru(0) atom and the two P atoms (maximum deviation of 0.0516 Šfrom the mean plane defined by the five constituent atoms). The coordinated C=C bond has a length of 1.449 (3) Å, which is significantly longer than that of a free C=C bond (1.34 Å). There are two C-H⋯π inter-actions involving neighbouring phenyl rings in the mol-ecule. In the crystal, mol-ecules are linked via two further C-H⋯π inter-actions.

Selective estrogen receptor modulator with estrogen does not affect the proliferation and apoptosis of uterine leiomyoma cells.

BACKGROUND: The administration of menopausal hormone therapy (MHT) in women with uterine leiomyomas is still debated. The purpose of this article is to study the proliferation and apoptosis of uterine leiomyoma cells under the impact of selective estrogen receptor modulator (SERM) combined with estrogen. METHODS: Primary cultured uterine leiomyoma cells in the perimenopausal period were treated with estrogen (17-beta estradiol) + SERM (raloxifene) as the tissue selective estrogen complex (TSEC) group, while both estrogen + medroxyprogesterone acetate (E+P) and estrogen (E) alone as were used as control groups. The expression of proliferating cell nuclear antigen (PCNA) and B-cell lymphoma-2 (Bcl-2) proteins was assessed by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and western-blot analysis, respectively. RESULTS: The proliferation in the TSEC group was weaker than the control groups (P<0.001). There was no statistical difference between the TSEC and blank control group on cell proliferation at 72 h (P=0.13). However, there was a significant difference between the other groups (P<0.001). PCNA expression of TSEC was lower than that of the E + P and E groups (P<0.05). There was no statistical difference in the expression of PCNA between the TSEC and blank control groups (P=0.63). Bcl-2 expression of TSEC was lower than that of the E + P and E groups (P<0.05). There was no statistical difference in the expression of Bcl-2 between the TSEC group and the blank control group (P=0.60). CONCLUSIONS: SERM combined with estrogen may have a better safety for perimenopausal women with uterine leiomyoma in MHT.

Effect of mixing intensity on hydrolysis and acidification of sewage sludge in two-stage anaerobic digestion: Characteristics of dissolved organic matter and the key microorganisms.

Mixing should be optimized in anaerobic digestion (AD) systems to achieve excellent biomaterials production in the sewage sludge (SS) management in wastewater treatment plant. AD depends on the coordinated activity of hydrolysis, acidification and methanogenesis. However, the effect of mixing intensity on characteristics of hydrolysis and acidification in AD of SS is still poorly understood. This study focused on the mixing intensity (30, 60, 90 and 120 rpm) effect on the characteristics of dissolved organic matter (DOM) and the key microorganisms in the hydrolysis and acidification of SS. Results showed that enhanced hydrolysis and acidification efficiency was obtained at mixing of 90 and 120 rpm (p < 0.05), while the maximum acetic acid (388 ±â€¯21 mg/L) was produced at 90 rpm. Mixing at 90 rpm enhanced the release of protein and polysaccharide as well as humic acid. Further analyses of DOM molecular features revealed that 90 rpm led to the highest molecular diversity and easily biodegradable molecules (lipid and proteins/amino sugars), which contributed to the maximum hydrolysis and acidification efficiency. Firmicutes and Actinobacteria significantly increased with mixing intensity (p<0.05), and Chloroflexi and Fusobacteria were enriched at mixing of 90 rpm, which favored the hydrolysis of SS. The enrichment of Clostridium XI and Clostridium sensu stricto contributed to the acidification of DOM at 90 and 120 rpm. The results of this study can advance our knowledge about mixing intensity effects on the AD systems of SS. This research also showed how increasing mixing intensity to a relatively high speed can enhance the hydrolysis and acidification efficiency of SS.

Cell membrane characteristics and microbial population distribution of MBBR and IFAS with different dissolved oxygen concentration.

This paper investigated the influences of different dissolved oxygen (DO) concentration (0.71-1.32, 2.13-3.02 and 4.31-5.16 mg/L) on cell membrane characteristics and microbial population distribution of moving biofilm reactors. Two representative reactors, i.e., moving bed biofilm reactors and integrated fixed-film activated sludge were operated. Results indicated that both DO concentration of 0.71-1.32 mg/L and 4.31-5.16 mg/L could increase membrane lipid mobile fraction (49.4%-67.4%) of the microbes, however, through prompting the synthesis of branched fatty acids and unsaturated fatty acids, respectively. For the biofilms, the abundance of Bacteroidetes decreased and Actinobacteria increased with the increase of DO levels. The lowest EfOM content and the highest microbial diversities (1.14-1.52) was observed at DO of 2.13-3.02 mg/L. Redundancy analysis showed that changes of DO levels could alter cell membrane properties and bacterial community structures, and subsequently significantly influenced effluent organic matter composition of moving biofilm reactors.

Effects of DO levels on surface force, cell membrane properties and microbial community dynamics of activated sludge.

In this paper, we employ atomic force microscopy (AFM), fluorescence recovery after photobleaching (FRAP) technique, phospholipid fatty acids (PLFA) and MiSeq analysis to study the effects of traditional dissolved oxygen (DO) levels (0.71-1.32mg/L, 2.13-3.02mg/L and 4.31-5.16mg/L) on surface force, cell membrane properties and microbial community dynamics of activated sludge. Results showed that low DO level enhanced the surface force and roughness of activated sludge; the medium DO level decreased cell membrane fluidity by reducing the synthesis of branched fatty acids in the cell membrane; high DO level resulted in the highest protein content in the effluent by EEM scanning. Abundance of Micropruina, Zoogloea and Nakamurella increased and Paracoccus and Rudaea decreased with the increase of DO levels. RDA analysis suggested that saturated fatty acids (SFA), anteiso-fatty acids (AFA) and iso-fatty acids (IFA) were closely related to effluent quality as well as some genera.

Dynamics and liver disease specific aspects of quality of life among patients with chronic liver disease in Yunnan, China.

BACKGROUND: Patients with chronic liver diseases (CLD) may have compromised health related quality of life (HRQoL). Hepatitis B virus (HBV) infection has long been the leading cause of CLD including liver cancer and cirrhosis. Knowledge on different symptom profiles of CLD should help in development of comprehensive treatment and patient care plans. OBJECTIVE: To access the facets of HRQoL in chronic liver diseases throughout their spectrum of severity. MATERIALS AND METHODS: A cross-sectional study was conducted in the First Affiliated Hospital of Kunming Medical University in Yunnan Province of China. Both out- and inpatients undergoing treatment protocols for different HBV related liver disease states were consecutively collected from December 2012 to June 2013. ANOVA was used to compare the mean scores of EQ-5D and chronic liver disease questionnaire (CLDQ) among 5 disease groups. The relationship between demographic variables predicting global CLDQ scores and the domains of CLDQ was analysed. RESULTS: A total of 1040 patients including 520 without complications, 91 with compensated cirrhosis, 198 with decompensated cirrhosis, 131 with HCC and 100 with liver failure were recruited. All domains of CLDQ, the means of EQ-5D value and EQ VAS exhibited significant decline with worsening of disease severity from uncomplicated HBV to liver failure. The multivariate regression demonstrated the reduction of mean scores of CLDQ domain at advanced stage. Patients with liver failure and HCC had more HRQoL impairment than other disease states. No effect of patient gender was found. Patient age was associated with 'fatigue' and 'worry' domains (p=0.006; p=0.004) but not with other domains and global scores of CLDQ and ED-5D. CONCLUSIONS: The HRQoL in chronic hepatitis B patients is greatly affected by disease states. Care for HBV-related diseases should consider not only the outcomes of treatment strategies but also improvement in patient wellbeing.