ABSTRACT
BACKGROUND: This study aimed to analysis the clinical characteristics and prognosis of acute STEMI in patients aged ≤ 45 years. METHODS: Seven hundred and one patients with STEMI from Liaocheng People's Hospital from January 2018 to March 2021 were included in this study. Clinical characteristics, management, and outcomes (average follow-up: 11.5 months) were compared between patients aged ≤ 45 years and those aged > 45 years. RESULTS: Of the patients with STEMI who underwent primary percutaneous coronary intervention, 108 (15.4%) were aged ≤ 45 years. Compared to the older group, the younger patient group included more males, current smokers, and those with alcohol use disorder (AUD) or a family history of ischaemic heart disease (IHD). The culprit vessel in young patients was the left anterior descending (LAD) artery (60% vs. 45.9%, P = 0.031), which may have been due to smoking (odds ratio, 3.5; 95% confidence interval: 1.12-10.98, P = 0.042). Additionally, young patients presented with higher low-density lipoprotein and lower high-density lipoprotein levels than older patients; uric acid levels were also significantly higher in younger patients than that in the older group. Diabetes showed a trend toward major adverse cardiovascular events (MACE) in both groups; age and sex were both independent predictors of MACE in older patients. CONCLUSION: More patients who were smokers, had AUD, or a family history of IHD were present in the young patient group. Hyperuricaemia (but not dyslipidaemia) was a prevalent risk factor in patients aged ≤ 45 years. Diabetes should be controlled to reduce cardiovascular events in young patients.
Subject(s)
Anterior Wall Myocardial Infarction , Coronary Artery Disease , Myocardial Ischemia , Percutaneous Coronary Intervention , ST Elevation Myocardial Infarction , Male , Humans , Aged , ST Elevation Myocardial Infarction/diagnosis , ST Elevation Myocardial Infarction/therapy , ST Elevation Myocardial Infarction/etiology , Retrospective Studies , Treatment Outcome , Risk Factors , Anterior Wall Myocardial Infarction/etiology , Coronary Artery Disease/etiology , Myocardial Ischemia/etiology , Percutaneous Coronary Intervention/adverse effects , Arrhythmias, Cardiac/etiologyABSTRACT
OBJECTIVE: Sepsis-associated encephalopathy (SAE) is a severe and common complication of sepsis and can induce cognitive dysfunction and apoptosis of neurons and neuroinflammation. Emodin has been confirmed to have anti-inflammatory effects. Thus, we sought to investigate the role of Emodin in SAE. METHODS: The cecal ligation and puncture (CLP) method was used for the establishment of SAE in mice model. For treatment of Emodin, intraperitoneal injection of 20 mg/kg Emodin was performed before the surgery. The Morris water maze and open field tests were carried for measurement of cognitive dysfunction. Hematoxylin and eosin staining was for histological analysis of hippocampus. Cell apoptosis of hippocampus neurons was measured by TUNEL staining. Pro-inflammatory and anti-inflammatory cytokines in hippocampus tissue homogenate were evaluated by ELISA. BDNF/TrkB signaling-related proteins (TrkB, p-TrkB, and BDNF), autophagy-related proteins (LC3 II/I and Beclin-1), and apoptosis-related proteins (Bax, Bcl-2, and cleaved caspase-3) were detected by Western blotting. RESULTS: Emodin significantly inhibited apoptosis and induced autophagy in hippocampal neurons of CLP-treated mice. In addition, Emodin significantly ameliorated CLP-induced cognitive dysfunction and pathological injury in mice. Meanwhile, Emodin notably inhibited CLP-induced inflammatory responses in mice via upregulation of BDNF/TrkB signaling, while the effect of Emodin was partially reversed in the presence of K252a (BDNF/TrkB signaling inhibitor). CONCLUSION: Emodin significantly inhibited the progression of SAE via mediation of BDNF/TrkB signaling. Thus, Emodin might serve as a new agent for SAE treatment.
Subject(s)
Emodin , Sepsis-Associated Encephalopathy , Animals , Apoptosis , Autophagy/physiology , Brain-Derived Neurotrophic Factor/metabolism , Brain-Derived Neurotrophic Factor/pharmacology , Emodin/metabolism , Emodin/pharmacology , Hippocampus/metabolism , Humans , Mice , Mice, Inbred C57BL , Receptor, trkB/metabolism , Sepsis-Associated Encephalopathy/drug therapy , Sepsis-Associated Encephalopathy/metabolismABSTRACT
Optically active Ć-amino alcohols are very useful chiral intermediates frequently used in the preparation of pharmaceutically active substances. Here, a novel cyclohexylamine oxidase (ArCHAO) was identified from the genome sequence of Arthrobacter sp. TYUT010-15 with the R-stereoselective deamination activity of Ć-amino alcohol. ArCHAO was cloned and successfully expressed in E. coli BL21, purified and characterized. Substrate-specific analysis revealed that ArCHAO has high activity (4.15 to 6.34Ć¢ĀĀ U mg-1 protein) and excellent enantioselectivity toward the tested Ć-amino alcohols. By using purified ArCHAO, a wide range of racemic Ć-amino alcohols were resolved, (S)-Ć-amino alcohols were obtained in >99 % ee. Deracemization of racemic Ć-amino alcohols was conducted by ArCHAO-catalyzed enantioselective deamination and transaminase-catalyzed enantioselective amination to afford (S)-Ć-amino alcohols in excellent conversion (78-94 %) and enantiomeric excess (>99 %). Preparative-scale deracemization was carried out with 50Ć¢ĀĀ mM (6.859Ć¢ĀĀ g L-1 ) racemic 2-amino-2-phenylethanol, (S)-2-amino-2-phenylethanol was obtained in 75 % isolated yield and >99 % ee.
Subject(s)
Amino Alcohols/metabolism , Oxidoreductases Acting on CH-NH Group Donors/metabolism , Transaminases/metabolism , Amino Alcohols/chemistry , Arthrobacter/enzymology , Biocatalysis , Molecular Structure , Oxidoreductases Acting on CH-NH Group Donors/genetics , Stereoisomerism , Transaminases/geneticsABSTRACT
OBJECTIVES: To screening of bacteria with cyclic amino alcohol deamination activity for enantioselective synthesis of chiral cyclic Ć-amino alcohols. RESULTS: A new strain named Arthrobacter sp. TYUT010-15 with the (R)-selective deamination activity of cyclic Ć-amino alcohol has been isolated from nature via a high throughput solid-phase screening method. The reaction conditions of TYUT010-15 were optimized. Using the resting cell of TYUT010-15 as the catalyst, kinetic resolution of trans-2-aminocyclopentanol, trans-2-aminocyclohexanol and cis-1-amino-2-indanol was carried out to afford (1S, 2S)-trans-2-aminocyclopentanol, (1S, 2S)-trans-2-aminocyclohexanol and (1R, 2S)-cis-1-amino-2-indanol in > 99% ee and 49.6-50% conversion. Four aromatic Ć-amino alcohols and two amines were also resolved, (S)-Ć-amino alcohols and (R)-amines were obtained in > 99% ee. Preparation experiment was conducted with 200Ā mM (23.2Ā g L-1) racemic trans-2-aminocyclohexanol, yielding the desired (1S, 2S)-trans-2-aminocyclohexanol in 40% isolated yield, > 99% ee and 5.8Ā g L-1 d-1 space time yields. CONCLUSIONS: This study provides a high throughput solid-phase method for screening of bacteria with cyclic amino alcohol deamination activity and a first example for practical preparation of chiral cyclic Ć-amino alcohol by Arthrobacter sp. TYUT010-15.
Subject(s)
Amino Alcohols , Bacteria/metabolism , High-Throughput Screening Assays/methods , Amines/analysis , Amines/chemistry , Amines/metabolism , Amino Alcohols/analysis , Amino Alcohols/chemistry , Amino Alcohols/metabolism , Arthrobacter/genetics , Arthrobacter/metabolism , Bacteria/genetics , Colorimetry , Deamination , Kinetics , Stereoisomerism , Substrate SpecificityABSTRACT
Low temperature solder (In-based, Sn-Bi, Sn-Zn) has great advantages in aerospace and through-hole technology assemblies in IBM mainframe due to its unique low temperature characteristics. The review evaluates the effects of alloying elements, rare earth elements and nanoparticles on the wettability, microstructure, mechanical properties and oxidation resistance of the low-temperature solders.
ABSTRACT
Low-cost, accurate high-risk human papillomavirus (HR-HPV) tests are needed for cervical cancer screening in limited-resource settings. More than 200 cervical cytological specimens from hospital patients were collected and analyzed for a real-world study. We evaluated the analytical and clinical performance of four widely used HR-HPV test (Tellgen, Hybribio, Liferiver, and Sansure) based on real-time polymerase chain reaction technology platforms, compared with the cobas test. Cervical intraepithelial neoplasia grade 2 or worse lesions (CIN2+) were set as the disease endpoint, and all the five HPV tests were performed with equal sensitivity (McNemar's test; P = 0.971) and specificity (McNemar's test; P = 0.953). All genotyping using the INNO-LiPA HPV test showed that HPV-16, -52, and -54 were the most common types among CIN2+ cases. Overall, the four HR-HPV tests analyzed appear to be as effective as the cobas HPV test in both agreement and clinical performance. Therefore, each of these low-cost HPV test kits could be implemented in limited-resource settings to accelerate the control of cervical cancer. However, we suggest that there is a need to further standardize and optimize testing around clinical sensitivity and specificity.
Subject(s)
Clinical Laboratory Techniques/standards , Early Detection of Cancer/methods , Health Resources , Papillomavirus Infections/diagnosis , Uterine Cervical Dysplasia/diagnosis , Uterine Cervical Neoplasms/diagnosis , Adult , Aged , Cervix Uteri/pathology , Cervix Uteri/virology , Clinical Laboratory Techniques/economics , Clinical Laboratory Techniques/methods , Early Detection of Cancer/instrumentation , Female , Humans , Middle Aged , Papillomaviridae/genetics , Papillomavirus Infections/virology , Reagent Kits, Diagnostic/economics , Reagent Kits, Diagnostic/standards , Sensitivity and Specificity , Uterine Cervical Neoplasms/virology , Young Adult , Uterine Cervical Dysplasia/virologyABSTRACT
High concentration restaurant oily wastewater from restaurants and food processing industries discharged into water environment usually results in environment pollution and inhibits the activity of microorganisms in biological wastewater treatment systems. In this study, 75 strains from oily sludge were isolated with oil degradation activity for edible oil-contained wastewater. Eight isolates were able to grow well in liquid cultures with edible oil as the sole carbon source and discovered with high efficient oil-degrading ability. Seven out of eight isolates were identified as Acinetobacter and one isolate as Kluyvera cryocrescens, based on their 16S rRNA gene sequences. Three highly efficient oil degrading bacteria (Acinetobacter dijkshoorniae LYC46-2, Kluyvera cryocrescens LYC50-1a and Acinetobacter pittii LYC73-4b) were selected and their degradation characteristic were examined, the results showed that the three isolates were effective under pH range from 7.0 to 10.0, and temperature from 25 to 35Ā Ā°C. For degradation of 2-4% (v/v) of vegetable oil, > 85% degradation percentage were obtained within 30Ā h. Degradation of the higher concentration oil (6-8%, v/v) result in 50-70% degradation percentage within 72Ā h, and the degradation percentage for the isolated strains were decreased about 50% for the degradation of 10% oil (< 45%) compared to 2% oil. Different type of oils were also tested, > 90% of degradation percentage were obtained by the three isolates, implied that these strains are capable of removing various oils efficiently. These results suggested that Acinetobacter dijkshoorniae LYC46-2, Kluyvera cryocrescens LYC50-1a and Acinetobacter pittii LYC73-4b are potential species could be efficiently used for high concentration restaurant oily wastewater treatment and might be applicable to a wastewater treatment system for the removal of oil.
Subject(s)
Bacteria/isolation & purification , Restaurants , Sewage/microbiology , Wastewater/microbiology , Acinetobacter/genetics , Acinetobacter/isolation & purification , Bacteria/genetics , Bacteria/growth & development , Biodegradation, Environmental , Hydrogen-Ion Concentration , Kluyvera/genetics , Kluyvera/isolation & purification , Phylogeny , Plant Oils , RNA, Ribosomal, 16S/genetics , TemperatureABSTRACT
OBJECTIVE: To study the effect of early intervention with lipoxin A4 (LXA4) on septic mice. METHODS: Healthy male Balb/c mice aged 6-8 weeks were randomly divided into sham-operation group, sepsis group, 1-hour intervention group (intervention at 1 hour after sepsis), and 6-hour intervention group (intervention at 6 hours after sepsis) (n=8 each). A sepsis model was prepared by cecal ligation and puncture. The intervention groups received LXA4 at 0.01 Āµg/g body weight 1 or 6 hours after the model was established. Blood was taken from eyeballs at 24 hours after operation. Peritoneal lavage fluid and liver and lung tissue samples were collected. The bacterial colonies of whole blood and peritoneal lavage fluid were counted by dilution plating. The serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) were determined by cytometric bead array. The serum level of high mobility group box-1 (HGMB1) was determined using ELISA. The percentages of macrophages and neutrophils in peritoneal lavage fluid were determined by flow cytometry. Paraffin sectioning and hematoxylin-eosin staining were performed for the liver and lung tissue samples to observe pathological damage. RESULTS: Compared with the sham-operation group, the sepsis group had a significantly decreased percentage of macrophages and a significantly increased percentage of neutrophils in peritoneal lavage fluid (P<0.05), as well as significantly increased serum levels of IL-6, TNF-α, MCP-1, and HMGB1 (P<0.05); in addition, the sepsis group showed more vacuolar degeneration, hepatocyte swelling, and inflammatory cell infiltration in liver tissue, and more capillary congestion, pulmonary septal thickening, inflammatory cell infiltration, and partial tissue destruction in lung tissue. Compared with the sepsis group, the 1-hour and 6-hour intervention groups had a significantly increased percentage of macrophages in peritoneal lavage fluid (P<0.05) and significantly reduced bacterial load in whole blood (P<0.05), serum levels of IL-6, TNF-α, MCP-1, and HMGB1 (P<0.05), and degree of liver and lung tissue damage and inflammatory cell infiltration, but there was no significant difference in the percentage of neutrophils and bacterial load in peritoneal lavage fluid (P>0.05). Compared with the 6-hour intervention group, the 1-hour intervention group had a significantly decreased serum level of HMGB1 (P<0.05), but there was no significant difference in other indicators between the two groups (P>0.05). CONCLUSIONS: Early intervention with LXA4 may attenuate liver and lung injuries in septic mice, which may be explained by the decrease in serum levels of IL-6, TNF-α, MCP-1, and HMGB1, and it also may reduce the bacterial dissemination in the whole blood of septic mice, which may be explained by the increase in the percentage of peritoneal macrophages.
Subject(s)
Sepsis , Animals , Early Intervention, Educational , Interleukin-6 , Lipoxins , Male , Mice , Tumor Necrosis Factor-alphaABSTRACT
OBJECTIVE: To study the protective effect of lipoxin A4 (LXA4) against sepsis induced by lipopolysaccharide (LPS) in rats with obesity and its effect on the expression of Toll-like receptor 4 (TLR4) and TNF receptor-associated factor 6 (TRAF6) in the liver. METHODS: A total of 60 male Sprague-Dawley rats aged three weeks were randomly divided into a normal group and an obesity group, with 30 rats in each group. A rat model of obesity was established by high-fat diet. Each of the two groups was further randomly divided into control group, sepsis group, and LXA4 group, and 8 rats were selected from each group. The rats in the control, sepsis, and LXA4 groups were treated with intraperitoneal injection of normal saline, LPS, and LXA4+LPS respectively. Twelve hours later, blood samples were collected from the heart and liver tissue samples were also collected. ELISA was used to measure the serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Western blot was used to measure the protein expression of TLR4 and TRAF6 in liver tissue. Quantitative real-time PCR was used to measure the mRNA expression of TLR4 and TRAF6. RESULTS: After being fed with high-fat diet for 6 weeks, the obesity group had significantly higher average weight and Lee's index than the normal group (P<0.05). Compared with the normal group, the obesity group had significant increases in the serum levels of IL-6 and TNF-α (P<0.05). In the normal group or the obesity group, the sepsis subgroup had significant increases in the serum levels of IL-6 and TNF-α compared with the control subgroup (P<0.05), while the LXA4 subgroup had significant reductions in the two indices compared with the sepsis subgroup (P<0.05). Compared with the normal group, the obesity group had significant increases in the protein and mRNA expression of TLR4 and TRAF6 (P<0.05). In the normal group or the obesity group, the sepsis subgroup had significant increases in the protein and mRNA expression of TLR4 and TRAF6 compared with the control subgroup (P<0.05). Compared with the sepsis subgroup, the LXA4 subgroup had significant reductions in the protein and mRNA expression of TLR4 and TRAF6 (P<0.05). CONCLUSIONS: LXA4 can reduce the serum levels of IL-6 and TNF-α and alleviate inflammatory response. LXA4 can inhibit the expression of TLR4 and TRAF6 in the liver of septic rats, possibly by inhibiting the TLR4 signaling pathway.
Subject(s)
Lipoxins/administration & dosage , Obesity/drug therapy , Sepsis/drug therapy , TNF Receptor-Associated Factor 6/genetics , Toll-Like Receptor 4/genetics , Animals , Humans , Interleukin-6/genetics , Interleukin-6/metabolism , Liver/drug effects , Liver/metabolism , Male , Obesity/complications , Obesity/genetics , Obesity/metabolism , Rats , Rats, Sprague-Dawley , Sepsis/complications , Sepsis/genetics , Sepsis/metabolism , Signal Transduction/drug effects , TNF Receptor-Associated Factor 6/metabolism , Toll-Like Receptor 4/metabolism , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolismABSTRACT
To study the expression and subcellular localization of recombinant dammarenediol-Ć¢Ā Ā” synthase (DS) in Saccharomyces cerevisiae, the dammarenediol-Ć¢Ā Ā” synthase gene ds was cloned from Panax ginseng, and the gene ds was fused with the gene of green fluorescent protein to obtain the fusion gene ds-gfp. The recombinant expression plasmids pESC-HIS-DS and pESC-HIS-DS-GFP were constructed and transformed into S. cerevisiae INVSc1 to obtain recombinant strains INVSc1-DS and INVSc1-DS-GFP. Microsomes of recombinant strains were prepared by differential centrifugation and observed by fluorescence microscope. The green fluorescence was only detected in INVSc1-DS-GFP microsomes, which indicated that DS was a membrane protein. It was also proved that dammarenediol-Ć¢Ā Ā” was produced from the cyclization of 2,3-oxidosqualene catalyzed by DS through in vitro enzymatic reaction. In addition, our results revealed that the fusion expression of ds with gfp significantly improved the production of dammarenediol-Ć¢Ā Ā” from 7.53 mgĀ·g(-1) to 12.24 mgĀ·g(-1). This study provides a new strategy in the optimization of the pathway of ginsenosides biosynthesis in S.cerevisiae.
Subject(s)
Alkyl and Aryl Transferases/genetics , Panax/enzymology , DNA, Complementary , Ginsenosides/biosynthesis , Recombinant Fusion Proteins/genetics , Saccharomyces cerevisiae , Saponins/biosynthesis , Squalene/analogs & derivatives , TriterpenesABSTRACT
Hypertriglyceridemia leads to liver steatosis, cardiovascular disease, and type 2 diabetes. Although HCBP6 (hepatitis C virus core-binding protein 6) was previously shown to be an HCV (hepatitis C virus) core-binding protein, its biological function remains unclear. Here, we demonstrate that HCBP6 negatively regulates intracellular triglyceride (TG) levels in hepatocytes. We found that bidirectional manipulation of hepatocyte HCBP6 expression by knockdown or overexpression results in increased or decreased TG accumulation, respectively. In addition, HCBP6 mRNA and protein levels exhibited significant time- and dose-dependent increases in a cellular model of lipid-overload hepatic steatosis. Furthermore, TG levels are regulated by HCBP6-sterol regulatory element binding protein 1c (SREBP1c)-mediated fatty acid synthase (FASN) expression. We also demonstrate that HCBP6 mRNA and protein expression is inhibited by microRNA-122 (miR-122), and miR-122 overexpression elicited more robust translational repression of luciferase activity driven by the full 3'-UTR of HCBP6. Taken together, our results provide new evidence that miR-122-regulated HCBP6 functions as a sensor protein to maintain intrahepatocyte TG levels.
Subject(s)
Fatty Acid Synthase, Type I/metabolism , Fatty Liver/genetics , MicroRNAs/biosynthesis , Sterol Regulatory Element Binding Protein 1/metabolism , Triglycerides/metabolism , Viral Core Proteins/metabolism , Fatty Acid Synthase, Type I/genetics , Fatty Liver/pathology , Fatty Liver/virology , Gene Expression Regulation , Hepacivirus/metabolism , Hepacivirus/pathogenicity , Hepatocytes/metabolism , Homeostasis , Humans , Liver/metabolism , Liver/pathology , Liver/virology , MicroRNAs/metabolism , RNA, Messenger/biosynthesis , Signal Transduction/genetics , Sterol Regulatory Element Binding Protein 1/genetics , Viral Core Proteins/geneticsABSTRACT
A series of phenylpiperazine derivatives (3a-3q) were designed and synthesized. In vitro assays indicated that several phenylpiperazine derivatives had excellent antiproliferative properties against four cancer cell lines including multidrug-resistant cancer cell lines, with IC50 values in the low micromolar range. The average IC50 of the most active compound 3b is 0.024ĀµM to the MCF-7 cell line. In addition, the mechanism of action of these new analogues was investigated by molecular docking studies, insulin-like growth factor 1-receptor (IGF-1R) kinase assay and apoptosis induced assay. These studies confirmed that these new phenylpiperazine derivatives maintain their mechanisms of action by disrupting IGF-1R kinase.
Subject(s)
Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Cell Proliferation/drug effects , Piperazines/chemistry , Piperazines/pharmacology , Receptor, IGF Type 1/antagonists & inhibitors , Apoptosis/drug effects , Breast Neoplasms/drug therapy , Breast Neoplasms/enzymology , Female , Humans , MCF-7 Cells , Molecular Docking Simulation , Protein Kinase Inhibitors/chemistry , Protein Kinase Inhibitors/pharmacology , Receptor, IGF Type 1/metabolismABSTRACT
Our knowledge about pathophysiology of intracerebral hemorrhage (ICH) mainly originates from preclinical models of ICH. In this study, cerebral ultrastructure surrounding hematoma and its correlation with clinical severity were investigated in ICH patients. Thirty patients with basal ganglia hemorrhage and 6 control subjects were enrolled. Surgical evacuation was performed for patients with a blood loss >30 ml. Stroke severity was assessed using the Glasgow Coma Scale (GCS) and the National Institute of Health Stroke Scale (NIHSS). Transmission electron microscopy (TEM) was used to evaluate the ultrastructural characteristics of tissue specimens. Neural cells surrounding the hematomas showed evidence of cell swelling and necrosis. Decreased numbers of organelles and mitochondrial cristae were accompanied by cytoplasmic vacuolization, nuclear membrane invagination and breakdown, and intranuclear chromatic agglutination. These changes resulted in disintegration together with malacia, disappearance of the nucleus and nucleolus, and karyopyknosis. More serious ultrastructural damage was seen in patients with greater NIHSS scores, lower GCS scores, and greater bleeding volumes (p < 0.001). These findings suggest that neural cells undergo unfavorable ultrastructural changes that are responsible for dysfunction after ICH.
Subject(s)
Basal Ganglia Hemorrhage/pathology , Brain/ultrastructure , Adult , Aged , Female , Glasgow Coma Scale , Hematoma/pathology , Humans , Male , Microscopy, Electron, Transmission , Middle Aged , Stroke/pathologyABSTRACT
The cyclization of 2,3-oxidosqualene is the key branch point of ergosterol and triterpenoid biosynthesis. Downregulation of 2,3-oxidosqualene metabolic flux to ergosterol in Saccharomyces cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway. In our study, primers were designed according to erg7 gene sequence of S. cerevisiae. Three fragments including 5' long fragment, 5' short fragment and erg7 coding region fragment were amplified by PCR. 5' long fragment consists of the promoter and a part of erg7 coding region sequence. 5' short fragment consists of a part of promoter and a part of erg7 coding region sequence. These fragments were inserted reversely into pESC-URA to construct antisense expression plasmids. The recombinant plasmids were transformed into S. cerevisiae INVSc1 and recombinant strains were screened on the nutritional deficient medium SD-URA. The erg7 expression level of recombinant strains, which harbored antisense expression plasmid of erg7 coding region, was similar to that of INVScl by semi-quantitative PCR detection. But erg7 expression level of recombinant strains, which harbored 5' long antisense fragment and 5' short antisense fragment, was significantly lower than that of the control. The results of TLC and HPLC showed that the ergosterol content of recombinant strains, which harbored 5' long antisense fragment, decreased obviously. The ergosterol contents of the others were almost equal to that of INVSc1. Lanosterol synthase gene expression was downregulated by antisense RNA technology in S. cerevisiae, which lays a foundation for reconstructing triterpenoid metabolic pathway in S. cerevisiae by synthetic biology technology.
Subject(s)
Intramolecular Transferases/metabolism , RNA, Antisense , Saccharomyces cerevisiae/genetics , Squalene/analogs & derivatives , DNA Primers , Down-Regulation , Gene Expression , Intramolecular Transferases/genetics , Plasmids , Polymerase Chain Reaction , Saccharomyces cerevisiae/enzymology , Squalene/metabolism , Transformation, GeneticABSTRACT
Pichia pastoris is one of the most important systems used in the field of molecular biology for the expression of recombinant proteins. The system has advantages of high expression, high stability, high secretion, easy high-density fermentation and low cost. Many factors affect the expression of recombinant protein, such as gene copy number, codon usage preference, type of promoter, molecular chaperones, glycosylation, signal peptide and fermentation process. In this review, research advances of the above aspects are summarized, which lay a foundation for improving the expression of recombinant proteins in P. pastoris.
Subject(s)
Fermentation , Pichia/metabolism , Recombinant Proteins/biosynthesis , Gene Dosage , Glycosylation , Molecular Chaperones , Promoter Regions, Genetic , Protein Sorting SignalsABSTRACT
Lanosterol synthase is encoded by the erg7 gene and catalyzes the cyclization of 2, 3-oxidosqualene, which is a rate-limiting step of the inherent mevalonate (MVA)/ergosterol metabolic pathway in Saccharomyces cerevisiae. The intermediate 2, 3-oxidosqualene is also the precursor of triterpenoids. Therefore, the cyclization of 2, 3-oxidosqualene is the key branch point of ergosterol and triterpenoids biosynthesis. Down-regulation of 2, 3-oxidosqualene metabolic flux to ergosterol in S. cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway reconstructed by the synthetic biology approach. To construct erg7 knockout cassette harboring the loxP-Marker-loxP element, long primers were designed, which were homologous to the sequences of both erg7 ORF and plasmid pUG66. The cassette was transformed into diploid wild strain INVSc1 by LiAc/SS Carrier DNA/PEG method and then erg7 gene haploid deficient mutant was obtained by homologous recombination. The results of semiquantitative PCR and real-time quantitative PCR revealed that erg7 expression level in erg7 gene haploid deficient mutant is one time lower than that in wild strain. The results of TLC and HPLC showed that the ergosterol content in deficient mutant decreased to 42% of that in wild strain.
Subject(s)
Ergosterol/metabolism , Intramolecular Transferases/genetics , Saccharomyces cerevisiae/genetics , Squalene/analogs & derivatives , Chromatography, High Pressure Liquid , DNA Primers , Down-Regulation , Haploidy , Polymerase Chain Reaction , Squalene/metabolismABSTRACT
The objective of the present study was to reveal different tolerance of peanut plants to Ca deficiency by determining Ca uptake and Fourier transform infrared spectral (FTIR) differences of two peanut cultivars grown in nutrition solution. Peanut cultivars LH11 and YZ9102 were selected. Seedlings at the first leaf stage were cultivated for 28 days in nutrient solution with 0, 0.01 and 2.0 mmol x L(-1) Ca treatments, respectively. The results showed that under 0 and 0.01 mmol x L(-1) Ca supply, YZ9102 did not show Ca deficiency symptoms and the plant biomass did not change, whereas LH11 exhibited shoot-tip necrosis, smaller plant size, more lateral branches, and plant dry matter weights decreased significantly. YZ9102 had higher plant Ca concentration and Ca accumulation than LH11. Besides, for LH11, Ca was mainly accumulated in roots, while for YZ9102 mainly in leaves. As compared with plants cultivated in 2.0 mol x L(-1) Ca nutrition, root, stem and leaf of LH11 plants under Ca deficiency stress showed higher transmittance at peaks 1 060, 1 380, 1 655, 2 922, and 3 420 cm(-1) in FTIR spectra, indicating that the contents of protein, sugar and lipid decreased obviously in LH11 plants in condition that Ca supply was limited. However, the FTIR spectra of YZ9102 were less affected by Ca deficiency. It is suggested that YZ9102 might be more tolerant to Ca deficiency.
Subject(s)
Arachis/physiology , Calcium/metabolism , Biomass , Plant Leaves , Plant Roots , Seedlings , Spectroscopy, Fourier Transform Infrared , Stress, PhysiologicalABSTRACT
BACKGROUND: Gelastic seizure (GS) is a rare type of epilepsy that most commonly appears in patients with hypothalamic hamartoma. It is rarely associated with other types of brain damage. This particular type of epilepsy is relatively rare and has few links to other brain lesions. Temporal lobe malacia is mostly caused by cerebral infarction or cerebral hemorrhage, which can lead to seizures. We report a case of GS in a woman with temporal lobe malacia which was reported for the first time in the literature. CASE SUMMARY: A 73-year-old female, diagnosed case of GS, presented with repetitive stereotyped laughter a month prior to presentation, happening multiple times daily and with each time lasting for 5-15s. Electroencephalogram displayed a focal seizure seen in the right temporal region. Magnetic resonance imaging head with contrast showed a right temporal lobe malacia. The patient was started on levetiracetam daily. The patient indicated that they had fully recovered and were not experiencing any recurrent or stereotyped laughter during their daily routines. These results remained consistent even after a one-year follow-up period. CONCLUSION: GS can be caused by temporal lobe malacia, which is an uncommon but potentially grave condition. The outcome of this present case exhibited the importance of the temporal lobe in the genesis of GS.
ABSTRACT
INTRODUCTION: Patients with underlying diseases do not respond adequately to vaccines. Thus, continued research on the effects of vaccination in patients with comorbidities is crucial to evaluate the necessity of vaccination in this population. This study assessed the protective effects of inactivated vaccines on the severity and prognosis of COVID-19 in patients with comorbidities. METHODOLOGY: A real-world retrospective cohort study was conducted from April 7, 2022, to June 6, 2022, at the Fudan University Pudong Medical Center. The collected data included demographic characteristics, symptoms, clinical severity, and outcomes of the COVID-19 patients. RESULTS: A total of 3,996 indigenous confirmed cases and asymptomatic infections with the Omicron variant were enrolled. Of these, 1322 (33.1%) patients had chronic comorbidities. Compared to others, COVID-19 patients with comorbidities were older, had lower vaccination rates, longer days of nucleic acid conversion and hospitalization, and a higher incidence of severe-critical illness and composite endpoint. Multivariable analyses suggested that in the comorbidity group, two-dose- (odds ratio [OR] 0.38, 95% CI 0.24-0.60; OR 0.20, 95% CI 0.08-0.51) and three-dose vaccinated patients (OR 0.26, 95% CI 0.14-0.47; OR 0.21, 95% CI 0.08-0.58) had a lower risk of aggravation and the composite endpoint; similar results were observed in the non-comorbidity group. CONCLUSION: Two or more doses of inactivated vaccines could prevent deterioration and poor prognosis in Omicron-infected patients, regardless of the presence of an underlying disease. Our findings support maximizing coverage with inactivated vaccines in highly vaccinated populations, such as those in China.
Subject(s)
COVID-19 Vaccines , COVID-19 , Comorbidity , SARS-CoV-2 , Vaccines, Inactivated , Humans , COVID-19/prevention & control , COVID-19/epidemiology , Male , Female , COVID-19 Vaccines/administration & dosage , COVID-19 Vaccines/immunology , Middle Aged , Retrospective Studies , Vaccines, Inactivated/administration & dosage , Vaccines, Inactivated/immunology , Adult , Aged , SARS-CoV-2/immunology , China/epidemiology , Vaccine Efficacy , Severity of Illness IndexABSTRACT
The controlled synthesis of Eu2(WO4)3 nanostructures with different morphologies, namely ellipsoidlike, rodlike, cubelike, rod-bundlelike, and mesoporous spindlike, has been successfully achieved by the cationic surfactant-CTAB (cetyltrimethylammoniumbromide)-microemulsion-mediated and anionic surfactant-SDS (sodium dodecyl sulfate)-microemulsion-mediated solvothermal method separately. Various comparison experiments showed that fundamental experimental parameters, such as the water content, reaction temperature and the type of surfactants played important roles in the morphological control of Eu2(WO4)3 nanostructures. The possible growth mechanisms of these nanocrystals were elucidated in detail.