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Objective:To investigate the role of long non-coding RNA nuclear paraspeckle assembly transcript 1 (Neat1) in pyroptosis of ultraviolet B (UVB)-induced human lens epithelial cells (LECs) and to explore the possible mechanism.Methods:The human lens epithelial cell line HLE-B3 was cultured in vitro, and cells at log phase were exposed to ultraviolet B for 0, 2, 4 and 8 hours, respectively.The expression of cysteine aspartic acid-specific protease-1 (caspase-1), a protein related to pyroptosis, was detected by Western blot.The relative expression level of Neat1 in cells after different irradiation durations was determined by real-time quantitative PCR.Cell viability was determined by the cell counting kit-8 (CCK-8) method to screen the optimal irradiation duration for UVB-induced LECs pyroptosis, which was finally determined to be 4 hours.HLE-B3 cells were divided into negative siRNA transfection group, siRNA Neat1 transfection group, negative siRNA transfection+ irradiation group and siRNA Neat1 transfection+ irradiation group, and were transfected with corresponding reagents for 24 hours.The negative siRNA transfection+ irradiation group and siRNA Neat1 transfection+ irradiation group were irradiated with UVB for 4 hours after transfection.The cell viability was detected by the CCK-8 method.The pyroptosis rate was detected by flow cytometry.The expression levels of caspase-1, gasdermin D (GSDMD) and nod-like receptor protein 3 (NLRP3) proteins were detected by Western blot.The concentration of interleukin (IL)-1β was detected by enzyme-linked immunosorbent assay (ELISA). Ultrastructural changes in HLE-B3 cells were observed under a transmission electron microscope. Results:The grayscale of caspase-1 protein bands increased with the extension of irradiation duration.The relative expression levels of caspase-1 protein at 0, 2, 4 and 8 hours of irradiation were 0.05±0.01, 0.25±0.07, 0.51±0.04 and 0.74±0.02, respectively, with a statistically significant overall difference ( F=168.223, P<0.001), and significant differences were found in paired comparisons (all at P<0.05). With prolonged irradiation, the relative expression level of Neat1 mRNA increased and the cell viability decreased, with statistically significant differences in paired comparisons (all at P<0.05). Compared with negative siRNA transfection group, the cell viability was increased in siRNA Neat1 transfection group and decreased in negative siRNA transfection+ irradiation group, with statistically significant differences (both at P<0.01). Compared with negative siRNA transfection+ irradiation group, the cell viability was increased in siRNA Neat1 transfection+ irradiation group, showing a statistically significant difference ( P<0.05). The pyroptosis rate was significantly lower in negative siRNA transfection group and siRNA Neat1 transfection+ irradiation group than in negative siRNA transfection+ irradiation group, and the differences were statistically significant (both at P<0.01). The relative expression levels of caspase-1, NLRP3 and GSDMD proteins in negative siRNA transfection+ irradiation group were higher than those in negative siRNA transfection group and siRNA Neat1 transfection+ irradiation group and the differences were statistically significant (all at P<0.01). The concentration of IL-1β was significantly higher in negative siRNA transfection+ irradiation group than in negative siRNA transfection group and siRNA Neat1 transfection+ irradiation group, and the differences were statistically significant (all at P<0.05). Cell swelling, formed cell membrane pores, vacuolated cells and fuzzy mitochondrial cristae were seen in negative siRNA transfection+ irradiation group and siRNA Neat1 transfection+ irradiation group by transmission electron microscopy.Compared with negative siRNA transfection+ irradiation group, slighter cell swelling, fewer cell membrane pores and lighter mitochondrial swelling were seen in siRNA Neat1 transfection+ irradiation group. Conclusions:Neat1 is involved in human LECs pyroptosis induced by UVB through the classic pyroptosis pathway mediated by caspase-1.Knockdown of Neat1 can inhibit the pyroptosis of human LECs.
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Objective:To analyse the pathogenic bacteria distribution and clinical characteristics of late-onset sepsis (LOS) among premature infants with gestational age less than 34 weeks in Henan Province.Methods:The clinical data of 6 590 premature infants admitted to 17 medical institutions in Henan Province from January 2019 to December 2020 were retrospectively analyzed. The gestational age of infants was less than 34 weeks and was admitted to the neonatal ward within 7 days after birth. SPSS 19.0 statistical software was used for data analysis.Results:Among 6 590 premature infants LOS developed in 751 cases (11.40%), of whom the diagnosis was confirmed in 276 cases (36.75%) and 475 cases (63.25%) were diagnosed clinically. The fatality rate related to LOS was 13.58%. There were significant differences in the incidence of LOS and infection-related mortality among infants with different gestational ages and body weights ( χ2=388.894 and 13.572, χ2=472.282 and 9.257, P<0.05 or <0.01). Among 276 children with confirmed LOS, 286 strains of pathogenic bacteria were isolated. Gram-negative bacteria were most prevalent (178 strains), accounting for 62.24% of all infections, followed by fungi (58 strains, 20.28%). Klebsiella pneumoniae was most frequently detected Gram-negative bacteria (117 strains, 40.91%), among which 32.48% (38/117) was carbapenem-resistant Klebsiella pneumoniae. The proportion of diagnosed sepsis, the proportion of catheterization, and the infection-related mortality of infants with LOS in tertiary hospitals were all higher than those in secondary hospitals ( χ2=6.212, 5.313 and 4.435, all P<0.05). The proportion of exclusive breastfeeding in secondary hospitals was lower than that in tertiary hospitals ( χ2=19.216, P<0.05). The time of antibacterial drug use before infection in specialized hospitals was longer than that in general hospitals ( χ2=3.276, P<0.05). Conclusion:The incidence of LOS among preterm infants in Henan Province is high, which was mainly caused by Gram-negative bacteria. The clinical characteristics of LOS caused by different pathogens and in different health institutions are different, the prevention and control strategy should be developed accordingly to reduce the incidence LOS of preterm premature infants.
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Objective:To analyze the characteristics of adults (non-pregnant women) infected with Listeria monocytogenes and to provide evidence for diagnosis and treatment of this disease. Methods:Patients admitted in West China Hospital of Sichuan University between January 2008 and April 2020 with Listeriadisease were enrolled in this study. The clinical data of patients including age, symptoms, underlying diseases and prognosis, as well as the laboratory results of WBC, neutrophil ratio (N%), procalcitonin (PCT), interleukin-6 (IL-6) and C-reactive protein (CRP), were obtained for analyzing the clinical infection and prognosis characteristics of the patients. Results:A total of 57 patients were included, patients were 18-83 (52.8±15.9) years old, percent of over 60 years old patients accounted for 35.09% (20/57). The incidence rate in summer season was the highest (45.61%, 26/57). There were 48 cases with comorbidities, sepsis occurred in 31 cases and meningitis in 18 cases. The time of diagnosis of Listeriosis was (6.1±2.9) days. After diagnosis, the target therapy was applied: such as carbapenem (26.32%, 15/57) and penicillins (22.81%, 13/57). The levels of WBC (6.77 [2.99, 9.54]×10 9/L vs 10.23 [6.71, 16.55]×10 9/L), NLR (3.07 [1.66, 8.16] vs 11.26 [5.66, 20.08]) and CRP (40.7 [16.9, 91.9] g/L vs 92.8 [59.9, 142.7] g/L) were significantly lower after treatment than those before treatment in 47 hospitalized patients (all P<0.05). Thirty-eight patients were treated according to the guidelines, 33 cases improved, 2 cases had poor prognosis and 2 cases died. Conclusion:The main risk factors of Listeria monocytogenes infection in adults (non-pregnant women) are underlying diseases, especially autoimmune diseases and pulmonary infections. Penicillins can be used as the first choice for empirical therapy. Carbapenems and erythromycin serve as the combination medications during the full course of treatment.
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Objective@#To evaluate the clinical characteristics of T-cell acute leukemia/lymphoma (T-ALL) and explore the prognosis significance of early T-cell precursor leukemia/lymphoma.@*Methods@#A cohort of 126 patients diagnosed with T-ALL from 2008 to 2014 in West China Hospital, Sichuan University were enrolled in this study. They were further categorized by immunophenotype according to the expression of T-cell lineage markers CD1a, CD8, CD5 and one or more stem cell or myeloid markers. The laboratory indicators and prognosis factors were also statistically analyzed.@*Results@#Of all patients, the ratio of male to female was 2.5∶1, with the median age of 25 years old (range 14 to 77) . The percentage of ETP-ALL was up to 47.6%. T-ALL patients showed higher ratio in first clinical remission rate (CR1) than T-LBL ones (64.4% vs 30.8%, P=0.032) . Group with WBC count higher than 50×109/L at presentation showed higher ration of achieving CR1 than those lower than 50×109/L (78.4% vs 50.9%, P=0.010) . In comparison with the non-ETP-ALL, ETP-ALL patients had older age of onset (P<0.001) , lower WBC count (P<0.001) , lower risk of CNS involvement (10.0% vs 30.2%, P=0.009) and slightly inferior overall survival (P=0.073) . T-cell lineage markers CD1a-, CD8- and CD4- positive patients had higher CR1 than their corresponding negative ones (P=0.002, P=0.000, P=0.001) , while CD33- and CD56- positive patients had lower ratio of achieving CR1 than their negative ones, respectively (P=0.035, P=0.035) .@*Conclusion@#Flow cytometry and associated markers for immunophenotyping was of significance in the diagnosis and prognosis monitoring of T-ALL/LBL. The percentage of ETP-ALL/LBL subtype was high in Chinese adolescent and adult T-ALL patients. ETP-ALL/LBL was a high risk subtype, which needs more precise standard for diagnosis and advanced therapies for better outcome.
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@#Objective To observe the effect of panaxadiol saponins (PDS) on intellectual decline and cerebral lesion induced by destabilization of cervical vertebra in mice.Methods44 mice were randomly divided into the normal control group, model group, high PDS doses group and low PDS doses group with 11 animals in each group. The model of destabilization of cervical vertebra was established by operating and intraperitoneal injection of PDS performed in the PDS high doses group (14 mg/kg) and low doses group (7 mg/kg) once everyday for 50 days. The memory ability of mice was evaluated by the water-maze test and tittup platform experiment. 50 days later, all mice were executed and the activities of superoxide dismutase (SOD), lactic dehydrogenase (LDH) and the content of maleic dialdehyde (MDA), nitric oxide (NO) in encephalon were tested.ResultsIn the PDS groups, the incubation period and error times in water-maze test shortened significantly ( P<0.05), aod the reaction period in tittup platform experiment shortened and wrong times decreased obviously ( P<0.01) compared with the model group. The activities of SOD, LDH increased ( P<0.01), and the content of MDA and NO decreased ( P<0.01, P<0.05) in the PDS group compared with the model group.ConclusionPDS has certain protective and improving effect on the decline of memory ability and cerebral lesion induced by destabilization of cervical vertebra in mice.