【Objective】 The study cloned prokaryotic expression of the CDs of dairy goat pituitary-specific transcription factor POU1F1 gene and analyzed the biological information.【Method】 The CDs of goat pituitary-specific transcription factor POU1F1 gene was amplified and cloned from total RNA of goat pituitary by RT-PCR according to the corresponding sequence in sheep.The biological information analysis was performed with it.The prokaryotic expression vector pET-32a-POU1F1 was constructed and expressed in E.coli BL21(DE3)pLysS.【Result】 The gene POU1F1 CDs is cloned and registered in GenBank(GenBank accession No.FJ547813).The ORF of POU1F1 gene consisted of 876 nucleotides encoding 291 amino acids with a POU-specific(POUs) domain from 124-198 amino acids and a Homeobox domain from 214-276 amino acids.The nucleotide sequence homology of POU1F1 CDs of Guanzhong dairy goat was found to be 98%,97%,91% and 86% compared with that of sheep(NM_001009350),bovine(NM_174579),human(NM_000306) and mouse(NM_008849),while the amino acid sequence homology was 98%,98%,96% and 92% respectively.The prokaryotic expression system of recombined vector pET-32a-POU1F1 was constructed successfully.The fusion protein His-POU1F1 was expressed by inducing in vitro.【Conclusion】 The functional amino acids coded by the gene POU1F1 are highly conserved in goat,sheep,cattle,human and mouse.It is implied that the gene POU1F1 in dairy goat shows the same function as it does in other species.
Hypoxia-induced cardiomyocyte apoptosis plays an important role in the development of ischemic heart disease. MicroRNAs (miRNAs or miRs) are emerging as critical regulators of hypoxia-induced cardiomyocyte apoptosis. miR-200c is an miRNA that has been reported to be related to apoptosis in various pathological processes; however, its role in hypoxia‑induced cardiomyocyte apoptosis remains unclear. In the present study, we aimed to investigate the potential role and underlying mechanism of miR-200c in regulating hypoxia‑induced cardiomyocyte apoptosis. We found that miR-200c was significantly upregulated by hypoxia in cardiomyocytes, as detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The lactate dehydrogenase, MTT, Annexin V/propidium iodide apoptosis and caspase-3 activity assays showed that downregulation of miR-200c markedly improved cell survival and suppressed the apoptosis of cardiomyocytes in response to hypoxia. Bioinformatics analysis and the dual-luciferase reporter assay demonstrated that miR-200c directly targeted the 3'-untranslated region of GATA-4, an important transcription factor for cardiomyocyte survival. RT-qPCR and western blot analysis showed that suppression of miR-200c significantly increased GATA-4 expression. Furthermore, downregulation of miR-200c upregulated the expression of the anti-apoptotic gene Bcl-2. However, the protective effects against hypoxia induced by the downregulation of miR‑200c were significantly abolished by GATA-4 knockdown. Taken together, our results suggest that downregulation of miR-200c protects cardiomyocytes from hypoxia-induced apoptosis by targeting GATA-4, providing a potential therapeutic molecular target for the treatment of ischemic heart disease.
Maximizing the pharmacological efficacy of albendazole (ABZ), an anti-echinococcosis drug, is essential in the long-term treatment of patients with echinococcosis. As a weakly alkaline drug, ABZ has a pH-dependent solubility that decreases dramatically from gastric fluid (pH 1.4) to intestinal fluid (pH 6.5), where it is absorbed. In this study, we endeavored to develop an optimized tablet formulation of ABZ to improve its dissolution and oral bioavailability from two aspects: a faster initial dissolution in the gastric pH condition (i.e., the "spring") and a more prolonged drug supersaturation in the intestinal pH condition (i.e., the "parachute"). To achieve this goal, ABZ-HCl salt was selected first, which demonstrated a higher intrinsic dissolution rate under pH 1.4 compared with the ABZ free base that is used in the commercial product Albenda. Second, by comparing the ABZ supersaturation kinetics under pH 6.5 in the presence of various polymers including poly(vinylpyrrolidone) (PVP), PVP/VA, hydroxypropyl methylcellulose (HPMC), and HPMC acetate succinate (HPMC-AS), HPMC-AS was found to be the most effective crystallization inhibitor for ABZ, likely due to the hydrophobic interaction between ABZ and HPMC-AS in an aqueous environment. The newly designed tablet formulation containing ABZ-HCl and HPMC-AS showed ∼3 times higher oral bioavailability compared with that of Albenda in Beagle dogs. More significantly, the anti-echinococcosis efficacy of the improved formulation was 2.4 times higher than that of Albenda in a secondary hepatic alveolar echinococcosis Sprague–Dawley rat model. The strategy of simultaneously improving the spring and parachute of an oral formulation of ABZ, by using a highly soluble salt and an effective polymeric crystallization inhibitor, was once again proven to be a viable and readily translatable approach to optimize the unsatisfactory oral medicines due to solubility and bioavailability limitations.
The infusion regulation of main quality component in tea was analyzed by using an oscillator to imitate the process of pouring boiling water into tea to study the effect of vibration after pouring boiling water into tea on quality of tea soup.The results showed that the process of pouring boiling water into tea could enhance the infusion rate of main quality components in tea soup,shorten the time to reach their highest concentration and have no obvious effect on the ratio of phenol/ammonia.
[Objective] To study the status of hMLH1 gene point mutations in gastric cancer families from northern China. [Methods] Blood samples from 94 non-cancerous members of four gastric cancer families and control individuals were collected. After DNA abstraction, the mutations of exon8 and exon12 of hMLH1 gene were investigated by PCR-SSCP-CE, followed by DNA sequencing. [Results] The mutant cases were 12 for exon8 and 10 for exon12 of the members in four gastric cancer families, while 2 for exon8 and 2 for exon12 in control individuals. The total mutant rate of the members being 23% in gastric cancer families was significantly higher than that being 4% in control individuals. Moreover, the mutant bases were the same as being found in hereditary nonpolyposis colorectal cancer (HNPCC). [Conclusion] The members of gastric cancer families from northern China may have similar genetic background as those of HNPCC.
Intervertebral disc degeneration (IDD), being the predominant root cause of lower back pain, has led to an enormous socioeconomic burden in the world. Ferroptosis is an iron-dependent nonapoptotic and nonpyroptotic programmed cell death associated with an increase in reactive oxygen species (ROS), which has been implicated in the pathogenesis of IDD. Activation transcription factor 3 (ATF3) is widely reported to promote ferroptosis and apoptosis in multiple diseases, but its roles and underlying regulatory mechanism in IDD have not been identified. FAoptosis is defined as a mixed cell death consisting of ferroptosis and apoptosis. The loss- and gain-of-function experiments demonstrated that ATF3 positively regulated tert-butyl hydroperoxide- (TBHP-) induced nucleus pulposus cell (NPC) FAoptosis, ROS production, inflammatory response, and extracellular matrix (ECM) degradation. Furthermore, silencing ATF3 ameliorated the progression of IDD in vivo, whereas its overexpression showed the opposite phenotype. Bioinformatics analysis and molecular experiments corroborated that ATF3 is a direct target of miR-874-3p, suggesting that the upregulation of ATF3 in IDD might be caused at least in part due to the downregulation of miR-874-3p in IDD, thereby relieving the inhibition of ATF3 by miR-874-3p. The findings revealed that ATF3 has the potential to be used as a promising therapeutic target against IDD.
The present study describes the case of a 77-year‑old female with a recently self-detected, painless, 7‑cm lump in the left breast, without evidence of metastasis clinically, who underwent mastectomy with dissection of the axillary lymph nodes. The tumor did not invade the chest wall and skin. The tumor was comprised of abundant chondroid matrix and fibrous tissue, with focal osteoid matrix, and was classified as a chondroblastic/fibroblastic variant. The tumor had a reverse zonal pattern. The tumor cells in the central portion were mainly spindle-like and sparse with minimal cytological atypia, while the remaining tumor cells in the periphery were mainly epithelioid, atypical and dense. Neoplastic osteoid woven bone or trabeculae were observed in the central portion of the tumor. No metastasis was identified in the axillary lymph nodes. The patient was alive without evidence of local recurrence or hematogenous spread at the 60-month follow-up.
Background. The competing endogenous RNA- (ceRNA-) mediated regulatory mechanisms are known to play a pivotal role in intervertebral disc degeneration (IDD). Our research intended to establish a ceRNA regulatory network related to IDD through bioinformatics analyses. Methods. The expression profiles of circRNA, miRNA, and mRNA were obtained from the public Gene Expression Omnibus (GEO) datasets. Then, we use sequence-based bioinformatics methods to select differentially expressed mRNAs (DEmRNAs), microRNAs (DEmiRNAs), or circRNAs (DEcircRNAs) related to IDD. We used ChEA3 to verify the targets of transcription factors (TFs). Then, we used DAVID to annotate the DEmRNAs. Finally, we constructed a potentially circRNA-miRNA-mRNA network related to IDD by predicting in the database (ENCORI, TargetScan, miRecords, miRmap, and circBank). Results. We identified 31 common DEmRNAs by Venn analysis, of which MMP2 was regarded as the key hub genes. Simultaneously, miR-423-5p and miR-185-5p were predicted as the upstream molecules of MMP2. Furthermore, a total of six DEcircRNAs were predicted as the upstream circRNAs of miR-423-5p and miR-185-5p. Then, a potential circRNA-miRNA-mRNA network related to IDD was constructed by bioinformatics analysis. Conclusion. A comprehensive ceRNA regulatory network was constructed, which was found to be significant in IDD progression.
In this study, a novel artificial intervertebral disc implant with modified "Bucklicrystal" structure was designed and 3D printed using thermoplastic polyurethane. The new implant has a unique auxetic structure with building blocks joined "face-to-face". The accompanied negative Poisson's ratio enables its excellent energy absorption and stability under compression. The deformation and load distribution behavior of the implant under various loading conditions (bending, torsion, extension and flexion) has been thoroughly evaluated through finite element method. Results show that, compared to natural intervertebral disc and conventional 3D implant, our new implant exhibits more effective stress transfer and attenuation under practical loading conditions. The implant's ability to contract laterally under compression can be potentially used to alleviate the symptoms of lumbar disc herniation. Finally, the biocompatibility of the implant was assessed in vitro and its ability to restore the physiological function of the disc segment was validated in vivo using an animal model.
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