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Objective:To analyze the effect on the expressions of nuclear factor-erythroid 2-related factor 2(Nrf2) and NAD(P)H quinone oxidoreductase 1 enzyme (NQO1) in A549 cells exposed to hyperoxia and interfered by small interfering RNA, and to investigate the role of Nrf2 and NQO1 in hyperoxia-induced lung injury as well as their relationship with apoptosis.Methods:A549 cells were gained by serial sub cultivation in vitro and then randomly divided into 4 groups: the air group without interference ( group Ⅰ), the hyperoxia group without interference (group Ⅱ), the air group transfected with Nrf2 siRNA (group Ⅲ), and the hyperoxia group transfected with Nrf2 siRNA (group Ⅳ). The hyperoxia groups (Ⅱ, Ⅳ group) were continuously exposed to an atmosphere containing a high concentration of oxygen (950 mL/L O 2, 50 mL/L CO 2), while the air groups (group Ⅰ, Ⅲ) were still placed in the incubator with 50 mL/L CO 2. In the pre-experiment, cells were transduced with a mixture of siRNA-1, siRNA-2, and siRNA-3. Then the siRNA with the highest efficiency for repressing Nrf2 expression was used for subsequent experiments. The mRNA and protein expression levels of Nrf2 and NQO1 in the 4 groups were detected by quantitative real-time polymerase chain reaction (qPCR) and Western blot.The distribution of Nrf2, Kelch-like ECH-associated protein 1(Keap1) and antioxidant response element(ARE) proteins in A549 cells after interference with Nrf2 was analyzed by immunofluorescence and confocal laser scanning microscope, and the cell apoptosis of the 4 groups were observed. Results:(1) Nrf2 siRNA significantly down-regulated the mRNA expression of Nrf2 in the groups siRNA-1, siRNA-2 and siRNA-3, and the inhibition efficiency of group siRNA-1 was the highest (80.57%). (2) The re-lative mRNA expression levels of Nrf2 and NQO1 in the group Ⅱ were 4.553±0.498 and 5.866±0.582, respectively.The mRNA and protein expression of Nrf2 and NQO1 and the cell apoptosis rate [(21.67±0.75)%]in the hype-roxia group were significantly higher than those in the group Ⅰ (all P<0.01). (3) The relative mRNA expression levels of Nrf2 and NQO1 in the group Ⅳ were 0.937±0.057 and 0.789±0.058, respectively.Compared with the group Ⅱ, the mRNA and protein expression of Nrf2 and NQO1 in the group Ⅳ were significantly decreased, while the cell apoptosis rate [(35.83±0.42)%]was significantly increased (all P<0.01). Conclusions:The abnormal expression of Nrf2 and NQO1 in A549 cells induced by hyperoxia and siRNA interference suggests that Nrf2 and NQO1 are involved in the pathogenesis of hyperoxia induced lung injury.Nrf2 and NQO1 are possibly protective factors in the hyperoxia induced lung injury and apoptosis.
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Internal control of public hospital can prevent and control risk exposure of public hospitals in their economic activities,in order to reasonably guarantee their legal compliance,safety and effective use of assets,as well as their authenticity and integrity of financial information.It can also effectively fend off fraud,corruption,and improve the quality of public health services and service efficiency.The authors analyzed the present situation and existing problems of internal control efforts at public hospitals in Ya'an city,and put forward countermeasures and suggestions as well.
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Three dimensional (3D) imaging and 3D printing have been applied in pancreatic malignancy management.The 3D imaging can not only be applied in pre-procedural evaluation and planning,but also it can be used for residents and fellows for training and education,and families of patients advsing.A model of pancreatic and its surrounding structures can be gotten by the 3D printing technique based on 3D imaging,which can not only recognize the anatomical structure,but also can make the guidance for the procedure.3D imaging and 3D printing technique have an important role in the management of pancreatic malignancy.In this review,we summarized the application of 3D) imaging and 3D printing in malignant neoplasm of pancreas.
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Objective@#Using CRISPR-Cas9 gene editing technology to achieve a number of genes co-deletion on the same chromosome.@*Methods@#CRISPR-Cas9 lentiviral plasmid that could induce deletion of Aloxe3-Alox12b-Alox8 cluster genes located on mouse 11B3 chromosome was constructed via molecular clone. HEK293T cells were transfected to package lentivirus of CRISPR or Cas9 cDNA, then mouse NIH3T3 cells were infected by lentivirus and genomic DNA of these cells was extracted. The deleted fragment was amplified by PCR, TA clone, Sanger sequencing and other techniques were used to confirm the deletion of Aloxe3-Alox12b-Alox8 cluster genes.@*Results@#The CRISPR-Cas9 lentiviral plasmid, which could induce deletion of Aloxe3-Alox12b-Alox8 cluster genes, was successfully constructed. Deletion of target chromosome fragment (Aloxe3-Alox12b-Alox8 cluster genes) was verified by PCR. The deletion of Aloxe3-Alox12b-Alox8 cluster genes was affirmed by TA clone, Sanger sequencing, and the breakpoint junctions of the CRISPR-Cas9 system mediate cutting events were accurately recombined, insertion mutation did not occur between two cleavage sites at all.@*Conclusion@#Large fragment deletion of Aloxe3-Alox12b-Alox8 cluster genes located on mouse chromosome 11B3 was successfully induced by CRISPR-Cas9 gene editing system.
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This study chiefly aimed at investigating the effects of Dan Zhi Jiang Tang Capsule (DJC) on myocardial Toll-like receptor 4 (TLR4),tumor necrosis factor α (TNFα) and interleukin-8 (IL-8) and the cardiopathologic changes in diabetic rats.Fifty male rats,excluding 8 rats in the control group,had been administered with high fat forage for 4 weeks,which established the diabetic rat model combined with the intraperitoneal injection of streptozotocin (STZ) at 35 mg· kg-1.The established model rats were randomly divided into the model group,the DJC high-dose group,the DJC low-dose group and the pioglitazone group.The rats in the model group and the control group were intragastrically administered with DJC at 1.08 and 0.54 g· kg-1 each day,while those of the pioglitazone group were treated with pioglitazone at 10 mg· kg-1 each day,and those of the model group and control group with saline of the same volume.The administrations lasted 8 weeks with the continues high-fat diet.The levels of TLR4,TNFα and IL-8 protein in myocardium of the rats were determined,and so were the blood glucose,HbA1c and blood lipids.Compared with the control group,the expressions of myocardial TLR4,TNFα and IL-8 in the model group were significantly increased (P＜ 0.01).The expressions of TLR4,TNFα and IL-8 in the DJC high-dose and low-dose groups were significantly decreased (P＜0.05 or P＜0.01).The cardiopathological report showed severe myocardial lesion in the model group.However,myocardial lesion in the DJC high-dose group was significantly mitigated,which presented better therapeutic effects than the DJC low-dose group and the pioglitazone group.The levels of blood glucose,HbA1c,TG,TC,LDL-C in the model group were significantly higher than those in the control group (P＜0.05 or P＜0.01).The levels of blood glucose,HbA1c,TG,TC and LDL-C were significantly decreased in the DJC high-dose and low-dose groups in comparison with the model group (P＜ 0.05 or P＜ 0.01).In conclusion,DJC down-regulated the expressions of TLR4,TNFα and IL-8 and improved the degree of myocardial lesion in the diabetic rats,showing favorable hypoglycemic and lipid-lowering effects.
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Intestine is responsible for the biotransformation of many orally-exposed chemicals. The constitutive androstane receptor (CAR/Nr1i3) is known to up-regulate many genes encoding drug-metabolizing enzymes and transporters (drug-processing genes/DPGs) in liver, but less is known regarding its effect in intestine. Sixty-day-old wild-type andmice were administered the CAR-ligand TCPOBOP or vehicle once daily for 4 days. In wild-type mice,mRNA was down-regulated by TCPOBOP in liver and duodenum.mice had altered basal intestinal expression of many DPGs in a section-specific manner. Consistent with the liver data (Aleksunes and Klaassen, 2012), TCPOBOP up-regulated many DPGs (, and) in specific sections of small intestine in a CAR-dependent manner. However, the mRNAs ofandwere previously known to be up-regulated by TCPOBOP in liver but were not altered in intestine. Interestingly, many known CAR-target genes were highest expressed in colon where CAR is minimally expressed, suggesting that additional regulators are involved in regulating their expression. In conclusion, CAR regulates the basal expression of many DPGs in intestine, and although many hepatic CAR-targeted DPGs wereCAR-targets in intestine, pharmacological activation of CAR in liver and intestine are not identical.
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Halohydrin dehalogenase is of great significance for biodegradation of the chlorinated pollutants, and also serves as an important biocatalyst in the synthesis of chiral pharmaceutical intermediates. A putative halohydrin dehalogenase (HheTM) gene from Tistrella mobilis KA081020-065 was cloned and over-expressed in Escherichia coli BL21 (DE3). The recombinant enzyme was purified by Ni-NTA column and characterized. Gel filtration and SDS-PAGE analysis showed that the native form of HheTM was a tetramer. It exhibited the highest activity at 50 degrees C. The nature and pH of the buffer had a great effect on its activity. The enzyme maintained high stability under the alkaline conditions and below 30 degrees C. HheTM catalyzed the transformation of ethyl(S)-4-chloro-3-hydroxybutyrate in the presence of cyanide, to give ethyl (R)-4-cyano-3-hydroxybutyrate, a key intermediate for the synthesis of atorvastatin.
Subject(s)
3-Hydroxybutyric Acid , Chemistry , Bacterial Proteins , Genetics , Metabolism , Cloning, Molecular , Escherichia coli , Hydrolases , Genetics , Metabolism , Hydroxybutyrates , Chemistry , Recombinant Proteins , Genetics , Metabolism , Rhodospirillaceae , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To evaluate the potential role of insulin-like growth factor-1 receptor mRNA (IGF-IR mRNA) in the onset and development of retinopathy in diabetic rats.</p><p><b>METHODS</b>A diabetic model was duplicated in Wistar rats. The early changes in the retina were examined using light and transmission electron microscopy. Expression of IGF-IR mRNA was analyzed using in situ hybridization.</p><p><b>RESULTS</b>Weak expression of IGF-IR mRNA (5%) was found in retinas of normal rats, but was significantly increased (15% and 18%) in the retinas of diabetic rats after 3 and 6 months of diabetes (P < 0.01). In situ hybridization and morphological study demonstrated that there was a positive correlation between IGF-IR mRNA expression and retinal changes at various stages.</p><p><b>CONCLUSION</b>Increased IGF-IR mRNA might play an important role in the onset and development of diabetic retinopathy.</p>
Subject(s)
Animals , Male , Rats , Blood Glucose , Diabetic Retinopathy , Glycated Hemoglobin , Insulin-Like Growth Factor I , Physiology , RNA, Messenger , Rats, Wistar , Receptor, IGF Type 1 , Genetics , Retina , Metabolism , PathologyABSTRACT
Objective To investigate the relation between the formaldehyde content of newly decorated houses and the time after decoration finished, which will offer an important reference to housing corporations and household. Methods The formaldehyde contents of newly decorated houses in Maoming were analyzed by colorimetry, Mar-Aug, 2003. Results The formaldehyde content in house A, in which the floor was compound floor boards, was higher than that in house B, in which the floor was wearable bricks. The formaldehyde content was not only related to the time after decoration finished, but also to the air temperature. After decoration finished, it must take half a year that the content of formaldehyde would meet the national standard limit. In consecutive testing, it was found that the content of formaldehyde would increase a little as the air temperature increased. Conclusion It must be attached importance to the formaldehyde content in the house floored with compound boards. It will need 6 months at least that the content of formaldehyde meet the requirement of the national standard after the newly house decoration.
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In TCM treatment of hepatitis B,one of refractory diseases,must follow strictly the principles of learning the past, but not rigidly adhere to it,and opening up and innovation. The principles of anti-virus, playing attention to objective detection. clinically re-researching, unifying disease differentiation and syndrome differentiation, as well as attaching importance to medicine toxicity and carefully selecting medicines and so on mentioned in this paper are a preliminary opinion based on the above view point in combination with long-term clinical practice and thinking.