Objective To determine enantiomeric impurity of etomidate using high performance liquid chromatography. Methods (R)-etomidate and (S)-etomidate were isopropanol-n-hexane. The flow rate of the mobile phase was 0.5mL/min. The detected wavelength was 242nm. Results (R)-etomidate and (S)-etomidate could be separated completely under these conditions. The precision of (R)-etomidate was 1.57% (n=3). The limit of detection of (R)-etomidate was 4.25ng/mL. The average percentage content of (S)-etomidate was 0.09% in the samples. Conclusion The method was repeatable and sufficiently sensitive to determine the enantiomeric impurity of etomidate. It allows the quantitation of the impurities at the 0.085% (w/w) level relative to etomidate at a concentration of the test solution of 5mg/mL.

Objective To quantify the expression levels of calpain 1 and caspase-3 in lichen planus (LP) lesions and their relationship with the apoptosis in keratinocytes.Methods Biopsy samples were obtained from the lesions of 20 patients with LP and normal skin of 10 healthy controls,and embedded in paraffin.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) was used to evaluate the apoptosis in keratinocytes,and immunohistochemical staining to detect the expressions of calpain 1 and caspase-3,in these tissue specimens.Data were processed by SPSS 13.0 software.Comparison analysis was carried out by t test for apoptosis index,and by rank sum test for the expressions of calpain 1 and caspase-3.Spearman correlation analysis was conducted to evaluate the relationship between these parameters.Results The apoptosis index of keratinocytes was higher in LP lesions than in the normal skin (67.59 ± 13.50 vs.28.26 ± 7.56,t =8.52,P ＜ 0.01).Significantly increased expressions of calpain 1 and caspase-3 were observed in the epidermis of LP lesions compared with the normal skin (T =78.00 and 77.00,respectively,both P ＜ 0.01).The expressions of both calpain 1 and caspase-3 were positively correlated with apoptosis index of keratinocytes (r =0.71 and 0.74,respectively,both P ＜ 0.01).Conclusions The expressions of calpain 1 and caspase-3 are upregulated in LP lesions,which may be closely associated with the accelerated apoptosis in keratinocytes.

Objective To investigate the effect of lncRNA CCAT1 and miR-130b-3p on the radiosensitivity of human pancreatic cancer cells PANC-1.Methods Real-time PCR was used to detect the relative expression levels of CCAT1 and miR-130b-3p in pancreatic cancer tissues and cell lines including PANC-1 cells irradiated with 2 Gy X-rays.After silencing CCAT1 and/or inhibiting miR-130b-3p expression,cell apoptosis rate,Caspase 3 activity and cell survival were detected by flow cytometry,Caspase 3 activity detection kit and colony formation assay,respectively.Cell survival curve was stimulated by the multi-target single-hit model.Based on the starBase v2.0 online analysis,the luciferase reporter gene assay,RNA-binding protein immunoprecipitation assay (RIP) and Real-time PCR assay were applied to verify the relationship between CCAT1 and miR-130b-3p.Results CCAT1 expression was up-regulated (t=6.322-8.555,P＜0.05),but miR-130b-3p expression was down-regulated (t =3.950-18.795,P＜ 0.05) in the radiation-resistant pancreatic cancer tissues,pancreatic cancer cell lines and 2 Gy-irradiated PANC-1 cells.When the CCAT1 silenced PANC-1 cells were irradiated with 2 Gy,cell survival fraction decreased (t=2.929,5.047,5.234,5.125,P＜0.05),apoptosis rate and Caspase 3 activity increased (t=6.953,6.836,P＜0.05).CCAT1 could selectively regulate miR-130b-3p expression.Inhibition of miR-130b-3p expression could enhance PANC-l cell survival (t =4.564,6.736,8.656,P＜0.05),but reduced apoptosis rate (t=5.234,P＜0.05) and Caspase 3 activity (t=10.440,P＜0.05).Conclusions Silencing CCAT1 promotes the expression of miR-130b-3p and enhances radiosensitivity of PANC-1 cells.

Objective To study the effects of simple portal hypertension on the endotoxin levels in serum and intestinal mucosa of rats. Methods A total of 16 rats were divided into the blank control group (4 rats) and the model groups (3-day group, 7-day group and 10-day group, 4 rats in each group). The rat model of partial portal vein ligation was established in the model groups, and samples of blood and jejunum, ileum and colon of the rats were taken on the 3rd, 7th and 10th days, respectively. Changes in the serum endotoxin levels were detected by ELISA. Histopathological changes of the intestinal tissues were examined by HE staining. Results The rat model of partial portal vein ligation was successfully established in all the model groups. The serum levels of endotoxin on the 3rd, 7th and 10th days in the model groups were not significantly different from that in the normal control group. Damages of different intestinal segments were not serious on the 3rd day after modeling. However, on the 7th day after modeling, there were some sowllen and damaged intestinal villi in the intestinal mucosa of each intestinal segment, and the connection between the epithelial cells and the lamina propria was broken, compared with that at 3 days after modeling. In addition, there were obvious damages in the intestinal mucosa and lamina propria on the 10th day, compared with that at 3 d and 7 d after modeling. Conclusions In the case of normal liver function, portal hypertension can cause intestinal mucosal damages within a short period of time, but the amount of endotoxin produced by intestine does not exceed the processing capacity of the liver and thus does not cause an increase in the serum endotoxin level.

Objective BALB/c mutant curly mice and normal BALB/c mice were genetically detected by microsatellite DNA marker analysis to detect the differential microsatellite loci between BALB/c mutant curly mice and normal mice.Methods 38 microsatellite DNA loci were selected and their variation in the BALB/c mutant curly mice, BALB/c mutant hairless mice and normal BALB/c mice were detected by multiplex fluorescence PCR and STR scanning genotyping.Results There were 27 the same microsatellite loci between the 38 microsatellite loci in BALB/c mutant curly mice and normal mice,and there were 11 differential loci, with a mutation rate of 28.9%(11/38). There were 30 the same sites between BABL/c mutant hairless mice and normal mice,and there were 8 different loci,with a mutation rate of 21.1%(8/38). There were also 12 differential loci between BABL/c mutant curly mice and hairless mice. Conclusions BALB/c mutant curly mice have a higher mutation rate and are significantly higher than those of hairless mice,demonstrating that the mutations in curly mice and hairless mice are two completely different mutations. These results provide reliable theoretical data for the future study and development of BALB/c mutant curly mice.

Objective:To investigate whether lncRNA LINC00909 affected the radiosensitivity of colorectal cancer cells by targeting miR-548-3p.Methods:The expression levels of LINC00909 and miR-584-3p in colorectal cancer and adjacent tissues were detected by qRT-PCR. The colorectal cancer cells SW480 and SW620 were cultured in vitro, and transfected with si-NC, si-LINC00909, miR-NC, miR-584-3p mimics, si-LINC00909, and anti-miR-NC and si-LINC00909, and anti-miR-584-3p, respectively. The cells were irradiated with a dose of 4 Gy. The cell survival fraction and sensitization enhancement ratio (SER) were detected by clone formation assay. Cell proliferation was detected by MTT assay. Cell migration and invasion were assessed by Trans well chamber assay. The targeting relationship between LINC00909 and miR-584-3p was confirmed by dual luciferase reporter assay. The effect of interfering with the expression of LINC00909 or inhibiting the expression of miR-584-3p on the weight of the xenograft tumor after irradiation was evaluated by subcutaneous xenograft experiment in nude mice. Results:The expression level of LINC00909 in colorectal cancer tissues was significantly up-regulated ( P<0.05), whereas the expression level of miR-584-3p was significantly down-regulated ( P<0.05). After interfering with the expression of LINC00909 or miR-584-3p overexpression, the cell survival fraction score was significantly reduced ( P<0.05), the SERs were 2.017 and 1.762, and cell proliferation, migration and invasion were suppressed (all P<0.05). Dual luciferase reporter assay confirmed that LINC00909 could target and bind to miR-584-3p. After interfering with the expression of LINC00909, the weight of the transplanted tumor was significantly reduced ( P<0.05), whereas the weight of the transplanted tumor was significantly increased after co-transfection with anti-miR-584-3p ( P<0.05). Conclusion:Interfering with the expression of LINC00909 can inhibit the proliferation, migration and invasion ability of colorectal cancer cells by up-regulating the expression of miR-548-3p, thereby enhancing the cell radiosensitivity.

Clostridia inhabiting in jiupei and pit mud plays key roles in the formation of flavour during the fermentation process of Luzhou-flavour baijiu. However, the differences of Clostridial communities between jiupei and pit mud remains unclear. Here, the species assembly, succession, and metabolic capacity of Clostridial communities between jiupei and pit mud were analysed by high-throughput sequencing and pure culture approaches. The ratio of Clostridial biomass to bacterial biomass in the pit mud was relatively stable (71.5%-91.2%) throughout the fermentation process. However, it varied widely in jiupei (0.9%-36.5%). The dominant Clostridial bacteria in jiupei were Clostridium (19.9%), Sedimentibacter (8.8%), and Hydrogenispora (7.2%), while Hydrogenispora (57.2%), Sedimentibacter (5.4%), and Caproiciproducens (4.9%) dominated in the Clostridial communities in pit mud. The structures of Clostridial community in pit mud and jiupei were significantly different (P=0.001) throughout fermentation. Isolated Clostridial strains showed different metabolic capacities of volatile fatty acids in pure culture. Spatial and temporal heterogeneity of Clostridial communities existed in the baijiu fermentation pit, which was closely related to the main flavour components of Luzhou-flavour baijiu.
Alcoholic Beverages ; microbiology ; Bacteria ; classification ; metabolism ; Clostridium ; physiology ; Fatty Acids, Volatile ; metabolism ; Fermentation ; Food Microbiology

Multi-species solid-state fermentation in a mud pit is one of the typical features of strong-flavor baijiu, in which archaea plays important roles, however, the archaeal community distribution and diversity during fermentation are still lack of research. The biomass, composition and succession of archaea communities in fermented grains and pit mud were analyzed by high throughput sequencing. The potential interaction between archaea and bacteria was analyzed by co-occurrence network. Results demonstrate that the average biomass of archaea in pit mud was about 200 times higher than that of fermented grains. There was no significant difference in archaeal community structure between fermented grains and pit mud (r=0.017, P=0.074), but succession patterns between them showed significant correlation (r=0.30, P=0.03). Methanobacterium was the most abundant archaea in fermented grains and pit mud, and other dominant groups included Methanosarcina, Methanocorpusculum, Methanoculleus, and Methanobrevibacter. The co-occurrence network analysis showed that Methanobacterium was positively correlated with most bacteria in fermented grains and pit mud, especially with Hydrogenispora and Caproiciproducens, the dominant bacteria in pit mud. Our results revealed the temporal and spatial distribution characteristics and potential functions of the archaeal community in the mud pit of strong-flavor baijiu.
Alcoholic Beverages/analysis* ; Archaea/genetics* ; Bacteria ; Fermentation ; Taste

Progressive functional deterioration in the cochlea is associated with age-related hearing loss (ARHL). However, the cellular and molecular basis underlying cochlear aging remains largely unknown. Here, we established a dynamic single-cell transcriptomic landscape of mouse cochlear aging, in which we characterized aging-associated transcriptomic changes in 27 different cochlear cell types across five different time points. Overall, our analysis pinpoints loss of proteostasis and elevated apoptosis as the hallmark features of cochlear aging, highlights unexpected age-related transcriptional fluctuations in intermediate cells localized in the stria vascularis (SV) and demonstrates that upregulation of endoplasmic reticulum (ER) chaperon protein HSP90AA1 mitigates ER stress-induced damages associated with aging. Our work suggests that targeting unfolded protein response pathways may help alleviate aging-related SV atrophy and hence delay the progression of ARHL.
Mice ; Animals ; Transcriptome ; Aging/metabolism* ; Cochlea ; Stria Vascularis ; Presbycusis