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Infective endocarditis (IE) is an infectious disease that affects the inner surface of the heart. Its first symptom often manifests as a localized neurological deficit, which can conceal the diagnosis of IE and delay the treatment. Here is a report of a severe case of IE with complicated central nervous system complications admitted to the First Hospital of Jilin University, so as to improve clinicians′ attention to the diagnosis and treatment of such conditions.
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β-N-acetylglucosaminidases (NAGases) can convert natural substrates such as chitin or chitosan to N-acetyl-β-D glucosamine (GlcNAc) monomer that is wildly used in medicine and agriculture. In this study, the BcNagZ gene from Bacillus coagulans DMS1 was cloned and expressed in Escherichia coli. The recombinant protein was secreted into the fermentation supernatant and the expression amount reached 0.76 mg/mL. The molecular mass of purified enzyme was 61.3 kDa, and the specific activity was 5.918 U/mg. The optimal temperature and pH of the BcNagZ were 75 °C and 5.5, respectively, and remained more than 85% residual activity after 30 min at 65 °C. The Mie constant Km was 0.23 mmol/L and the Vmax was 0.043 1 mmol/(L·min). The recombinant BcNagZ could hydrolyze colloidal chitin to obtain trace amounts of GlcNAc, and hydrolyze disaccharides to monosaccharide. Combining with the reported exochitinase AMcase, BcNagZ could produce GlcNAc from hydrolysis of colloidal chitin with a yield over 86.93%.
Subject(s)
Acetylglucosamine , Acetylglucosaminidase , Bacillus coagulans , Chitin , Chitinases , Hydrogen-Ion Concentration , Recombinant Proteins/geneticsABSTRACT
Objective To investigate the rebleeding rate after endoscopic selective variceal devascularization (ESVD) and the predictive factors for rebleeding in patients with hepatitis B cirrhosis and esophageal variceal bleeding (EVB). Methods The patients with hepatitis B cirrhosis and EVB who attended Beijing Ditan Hospital, Capital Medical University, from October 2010 to December 2019 and underwent ESVD for the first time were enrolled, and a total of 442 patients were screened out based on inclusion and exclusion criteria. Routine clinical indices, laboratory markers, imaging findings, and endoscopic findings were compared between patients, and the patients were followed up to observe rebleeding. The t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. The Kaplan-Meier method was used to describe rebleeding and survival status, and a Cox regression analysis was used to determine the independent risk factors for variceal rebleeding. Results The 1-, 2-, 3-, 4-, and 5-year cumulative rebleeding rates after first ESVD treatment were 25.11%, 33.94%, 39.82%, 42.08%, and 45.02%, respectively. The univariate analysis showed that age, systolic pressure, duration of antiviral therapy ≥1 year, ascites, white blood cell count, neutrophil, and direct bilirubin were associated with rebleeding (all P < 0.05), and the multivariate analysis showed that duration of antiviral therapy ≥1 year (hazard ratio [ HR ]=0.504, 95% confidence interval [ CI ]: 0.357-0.711, P < 0.001) and ascites ( HR =1.424, 95% CI : 1.184-1.714, P < 0.001) were independent influencing factors for variceal rebleeding. Conclusion ESVD has a low rebleeding rate in the treatment of hepatitis B cirrhosis with EVB, and presence of ascites and a short duration of antiviral therapy are independent risk factors for rebleeding after treatment.
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Objective:To study the change in serum fibroblast growth factor 21(FGF21)level and its correlation with liver functions in elderly patients with hepatitis B virus(HBV)-related cirrhosis.Methods:A total of 150 elderly patients with HBV infection admitted to Beijing Ditan Hospital and China-Japan Friendship Hospital from January 2019 to December 2020 were selected and divided into chronic hepatitis B(CHB)group(n=70)and HBV-related cirrhosis group(n=80). Healthy subjects were selected as the control group(n=50). Serum FGF21 was determined by ELISA method.Clinical data, clinical laboratory indicators and FGF21 as a core parameter of this study were collected and compared among control group, CHB group, HBV-related cirrhosis group.The correlations of FGF21 level with several liver function indexes were analyzed by Pearson correlation analysis.Results:The levels of total bilirubin(TBil)and total biliary acid(TBA)were significantly higher in HBV-related cirrhosis group than in CHB group.The levels of aspartate transaminase(AST), alanine transaminase(ALT), glutamyl transpeptadase(GGT), serum albumin, cholinesterase, FGF21 and HBV-DNA were significantly lower in HBV-related cirrhosis group than in CHB group(all P<0.05). The levels of serum FGF21 were(108.6 ± 7.3)ng/L, (92.5 ± 7.6)ng/L and(75.8 ± 6.3)ng/L in Child-Pugh A, B and C patients, respectively.The level of FGF21 was statistically significantly decreased with the increase of Child-Pugh grading( F=18.290, P=0.000). Serum FGF21 level was positively correlated with AST, ALT, TBil, cholin-esterase and albumin levels( r=0.652, 0.579, 0.609, 0.558, 0.613, all P<0.05). Conclusions:The level of serum FGF21 is significantly decreased in elderly patients with HBV-related cirrhosis, and is positively correlated with liver function indexes.
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Background@#Prolonged electrocardiogram (ECG) QRS duration has been associated with increased cardiovascular risk. It is unclear whether the main predictor of cardiovascular risk, the Framingham risk score also predicts short-term changes in ECG QRS duration. Our aim is to determine whether baseline Framingham risk score is associated with baseline or changes in QRS duration. @*Methods@#A retrospective cross-sectional analysis was performed using observational data obtained from two hundred two participants. Framingham risk score was calculated using an online risk calculator. QRS duration was obtained using a 10 s trace from a Welch Allyn PC-based 12-lead ECG system. @*Results@#Average follow-up duration was 3.3 ± 1.1 years. Mean QRS change was 1.8 ± 11.4 ms. Specifically, among two hundred two participants, there are 104 subjects with a greater QRS duration at follow-up, while 98 subjects had the same or a shorter follow-up QRS duration. Baseline Framingham risk score did not significantly predict an increase in QRSd with an odds ratio of 1.04 (P = 0.230). Regression analysis of QRS duration at baseline and Framingham risk at baseline had a weak association (R2= 0.020;P = 0.043). The Framingham risk score at follow-up was likewise has a weak association with follow-up QRS duration (R2= 0.045; P = 0.002). @*Conclusions@#Our results do not demonstrate a statistically significant association between Framingham risk parameters and future QRS duration changes over longitudinal time. QRS duration had variable changes between baseline and follow-up. This might suggest that a longer period of follow-up is required to document more stable increases in QRS duration associated with ventricular pathology. A larger population study is needed to confirm our observations.
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Background@#Prolonged electrocardiogram (ECG) QRS duration has been associated with increased cardiovascular risk. It is unclear whether the main predictor of cardiovascular risk, the Framingham risk score also predicts short-term changes in ECG QRS duration. Our aim is to determine whether baseline Framingham risk score is associated with baseline or changes in QRS duration. @*Methods@#A retrospective cross-sectional analysis was performed using observational data obtained from two hundred two participants. Framingham risk score was calculated using an online risk calculator. QRS duration was obtained using a 10 s trace from a Welch Allyn PC-based 12-lead ECG system. @*Results@#Average follow-up duration was 3.3 ± 1.1 years. Mean QRS change was 1.8 ± 11.4 ms. Specifically, among two hundred two participants, there are 104 subjects with a greater QRS duration at follow-up, while 98 subjects had the same or a shorter follow-up QRS duration. Baseline Framingham risk score did not significantly predict an increase in QRSd with an odds ratio of 1.04 (P = 0.230). Regression analysis of QRS duration at baseline and Framingham risk at baseline had a weak association (R2= 0.020;P = 0.043). The Framingham risk score at follow-up was likewise has a weak association with follow-up QRS duration (R2= 0.045; P = 0.002). @*Conclusions@#Our results do not demonstrate a statistically significant association between Framingham risk parameters and future QRS duration changes over longitudinal time. QRS duration had variable changes between baseline and follow-up. This might suggest that a longer period of follow-up is required to document more stable increases in QRS duration associated with ventricular pathology. A larger population study is needed to confirm our observations.
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Zearalenone (ZEN) and its derivatives are non-steroidal estrogenic mycotoxins mainly produced by Fusarium species. They are widely distributed in grain feeds originated from maize, barley, wheat and sorghum, causing serious harm to animal and human health. Currently, there is a pressing need of an efficient technology for ZEN degradation and detoxification. Because traditional physical and chemical methods could not effectively detoxify ZEN in grains, and might also affect the grain nutrients and food taste, and even result in secondary pollution, the biological technologies are developed to detoxify ZEN and its derivatives. In this paper, we reviewed the structure of ZEN and its derivatives, the fungi and bacteria species with ability of degradation of ZEN. In addition, the characterization, protein sequences and conformation of currently identified ZEN degrading enzymes, the only solved ZHD structure from Clonostachys rose were analyzed and compared, and the enzymes heterologous expression and application were also reviewed. This review will provide reference for reducing the cost of ZEN degrading enzymes by biological technologies such as enzyme engineering and fermentation engineering.
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Bacillus subtilis is Gram-positive aerobic bacterium and widely used as a heterologous protein expression host because of its safety and high protein secretion property. However, comparing to Escherichia coli, the low transformation efficiency limits the application of B. subtilis as a host cell for directed evolution of heterologous enzymes. Therefore, we optimized the competent cell preparation conditions for conventional plasmid, including the alteration of the medium, the concentration of inducer, the plasmid type, and other parameters. Compared with the original LB medium, YN medium improved the transformation efficiency by about 4 folds. The transformation efficiency enhanced by about 2 folds under induction with 1.5% xylose for 2 h. In addition, with plasmids prepared from E. coli GM272 strain the transformation efficiency increased by about 3 folds. Combining all these findings, the transformation efficiency of pDG1730 plasmid under the optimized conditions could reach 10⁶ CFU/μg, which was 2 orders of magnitude higher than that the original. Our findings provide references for directed evolution of enzymes and metabolic engineering in Bacillus subtilis.
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Bacillus subtilis is a generally recognized as safe (GRAS) strain that has been widely used in industries including fodder, food, and biological control. In addition, B. subtilis expression system also plays a significant role in the production of industrial enzymes. However, its application is limited by its low sporulation frequency and transformation efficiency. Immense studies have been done on interpreting the molecular mechanisms of sporulation and competence development, whereas only few of them were focused on improving sporulation frequency and transformation efficiency of B. subtilis by genetic modification. The main challenge is that sporulation and competence development, as the two major developmental events in the stationary phase of B. subtilis, are regulated by the complicated intracellular genetic regulatory systems. In addition, mutual regulatory mechanisms also exist in these two developmental events. With the development of genetic and metabolic engineering, constructing genetic regulatory networks is currently one of the most attractive research fields, together with the genetic information of cell growth, metabolism, and development, to guide the industrial application. In this review, the mechanisms of sporulation and competence development of B. subtilis, their interactions, and the genetic regulation of cell growth were interpreted. In addition, the roles of these regulatory networks in guiding basic and applied research of B. subtilis and its related species were discussed.
Subject(s)
Bacillus subtilis , Genetics , Physiology , Gene Regulatory Networks , Metabolic Engineering , Spores, Bacterial , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To analyze the association of micoRNA-related genes DROSHA single nucleotide polymorphisms (SNP) rs10719 and rs6877842, DICER1 rs3742330and GEMIN4 rs3744741 with prognosis of T-cell lymphoma.</p><p><b>METHODS</b>Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to determine the genotypes of the above 4SNPs and their associations with complete remission (CR) rate and overall survival (OS) in 163 patients with TCL.</p><p><b>RESULTS</b>Patients carrying the rs6877842 CG genotype had a significantly higher CR rate compared with those carrying the CC genotype (OR=0.07, 95% CI 0.01-0.72, P=0.026); the same for patients carrying the DICER1 rs3742330 GG genotype compared with those carrying the GA genotype (OR=0.15, 95% CI 0.02-0.97, P=0.047) or the AA genotype (OR=0.11, 95% CI 0.02-0.71, P=0.020). In addition, patients with the DICER1 rs3742330 GG genotype had a significantly improved OS compared with those carrying the GA (HR=9.02, 95% CI 1.22-66.92, P=0.031) or AA genotype (HR=8.77, 95% CI 1.19-64.67, P=0.033). The other two SNPs of rs10719 and rs3744741 had no significant association with CR or OS.</p><p><b>CONCLUSION</b>DROSHA rs6877842 and DICER1 rs3742330 were independent factors for TCL CR, and DICER1 rs3742330 was also an independent prognostic factor for TCL OS.</p>
Subject(s)
Humans , DEAD-box RNA Helicases , Genetics , Genetic Predisposition to Disease , Genotype , Lymphoma, T-Cell , Diagnosis , Genetics , MicroRNAs , Genetics , Minor Histocompatibility Antigens , Polymerase Chain Reaction , Polymorphism, Single Nucleotide , Prognosis , Ribonuclease III , Genetics , Ribonucleoproteins, Small Nuclear , GeneticsABSTRACT
Objective To investigate the expression of tissue factor and explore its clinical significances in pulmonary artery after acute pulmonary thromboembolism.Methods Thirty-four Japanese white rabbits (Level Ⅱ animals) were randomly (random number) assigned into four groups:group A (specimen of pulmonary artery was taken 3 hours after pulmonary embolism,n =8),group B (specimen of pulmonary artery was taken 8 hours after pulmonary embolism,n =8),group C (specimen of pulmonary artery was taken 24 hours after pulmonary embolism,n =8) and control group (pseudo-operations were carried out without injecting autologous blood clots,n =10).The animal model of pulmonary thromboembolism was established by injecting autologous blood clots into jugular vein through a 5F catheter and confirmed by digital subtraction angiography.The mRNA expression of TF in different parts of pulmonary artery was assayed by RT-PCR.The q test was utilized if there was a significant difference in a given continuous variable among three groups analyzed by ANOVA.Results The TF expression in the specimen adjacentto emboli was stable at 3 h,8 h or 24 hours after embolism.The mRNA expression of TF at 3 h and 8 h after embolism was lower in specimen taken from distal-end of morbid pulmonary artery than those adjacent to emboli.While at 24 hours after embolism,there were similar mRNA expressions in specimen either adjacent or distal to emboli.Conclusions The high expression of tissue factor in pulmonary artery tissue adjacent to emboli could lead to locally increased coagulation activity,indicating the necessity of initiating anti-coagulation treatment as soon as possible after acute pulmonary embolism.
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In this study, we efficiently expressed the active antimicrobial peptide (CAD), which fused with the site-mutated coat protein (EDDIE) of the classical swine fever virus, in Escherichia coli. First, we obtained the e-cad fusion gene from the CAD gene and the EDDIE gene using overlapping PCR. Then to get the recombinant expression vector (pETED), the e-cad fusion gene was cloned into the pET30a vector by a site-directed homologous recombination technique. The EDDIE-CAD fusion protein expressed in E. coli as inclusion bodies, and its yield was more than 40% of total bacterial proteins. After renaturated in vitro and self-cleavage of the fusion protein, we obtained the antimicrobial peptide Cecropin AD. Antimicrobial experiments showed that the Cecropin AD efficiently inhibited the growth of G+ and G- bacteria, but it weakly inhibited the growth of Saccharomyces. This method provides an excellent way for high expression of antimicrobial peptides when fused with EDDIE.
Subject(s)
Anti-Infective Agents , Metabolism , Capsid Proteins , Genetics , Metabolism , Cecropins , Genetics , Classical Swine Fever Virus , Genetics , Metabolism , Escherichia coli , Genetics , Metabolism , Genetic Vectors , Genetics , Mutation , Recombinant Fusion Proteins , Genetics , PharmacologyABSTRACT
OBJECTIVE To study Pseudomonas aeruginosa infection in respiratory tract of severe neurosurgery patients after tracheotomy,and discuss the prevention and management of nosocomial infection.METHODS The data of 19 severe patients admitted in neurosurgery from Dec 2005 to Oct 2007 who underwent tracheotomy were analyzed retrospectively.RESULTS Respiratory tract infection occurred in all 19 cases,the incidence rate was 100%.Eleven species and 73 strains of pathogenic bacteria were found.Thirty(41.10%)strains of P.aeruginosa were found in 12(63.16%)cases,the drug resistance rate was higher.CONCLUSIONS P.aeruginosa is the most important pathogenic bacteriaum of respiratory tract nosocomial infection in severe neurosurgery patients after tracheotomy.Strengthening the sterilization,isolation and airway management and selecting antibiotics based on drug sensitive test are the important methods to prevent and control infection.
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<p><b>OBJECTIVE</b>To investigate the association between the plasminogen activator inhibitor-1 (PAI-1) 4G/5G gene polymorphism and the occurrence of myocardial and cerebrovascular infarctions in individuals from Tianjin, China.</p><p><b>METHODS</b>The PAI-1 genotype was determined using allele-specific polymerase chain reaction (AS-PCR) in 56 myocardial infarction (MI) patients, 54 cerebrovascular infarction (CI) patients and 83 unrelated healthy controls. All subjects' clinical features and plasma PAI-1 activity levels were determined.</p><p><b>RESULTS</b>The PAI-1 genotype distribution frequency of the single guanine deletion/insertion 4G/5G polymorphism (located -675 bp upstream from the start of transcription) significantly differed between the patients and healthy controls. In the MI group, the 4G/4G-genotype frequency was increased, but the 4G/5G-genotype is decreased when compared to the control group. In the CI group, both the 4G/4G- and 4G/5G -genotypes occurred at a lower frequency than those in the control group (P < 0.001). The plasma PAI-1 activity level in the MI group was lowered as the presence of the 4G allele decreases. In the CI group, the frequency of 5G/5G was much higher than that of the control group (P < 0.001). The plasma PAI-1 activity level in the CI group was elevated as the presence of the 5G allele increased. Furthermore, positive correlation between triglyceride, glucose levels and PAI-1 activity were found in all three groups (P < 0.001).</p><p><b>CONCLUSIONS</b>The PAI-1 4G/5G gene polymorphism is associated with a higher risk of MI and CI in individuals in Tianjin, China. The deletion/insertion polymorphism is probably an important hereditary risk factor for heart diseases. Moreover, triglyceride and glucose levels of plasma have functional importance in regulating PAI-1 activity.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Asian People , Genetics , Cerebral Infarction , Genetics , China , Epidemiology , Gene Deletion , Myocardial Infarction , Genetics , Plasminogen Activator Inhibitor 1 , Genetics , Polymorphism, GeneticABSTRACT
Objective: To analyze the expression of CMTM1-v17 in normal prostate tissue and prostate carcinoma originated cell lines, and study its impact on the transactivation of androgen receptor and the possible mechanism. Methods: The expression of CMTM1-v17 in normal prostate tissue was analyzed with immunohistochemistry method. In immounocytochemistry was used to analyze the expression of CMTM1-v17 in prostate carcinoma originated cell lines. Luciferase assay was used to study the impact of CMTM1-v17 on the transactivation of AR and its mechanism. Results: The results of immunohistochemistry showed that CMTM1-v17 was highly expressed in prostate. In prostate cancer originated cell lines, CMTM1-v17 could also be detected in prostate cancer originated cell lines PC3, Du145 and LNCaP. And the results of luciferase implied that the relative luciferase activity of the PC3 cells transfected with 1 ?g and 2 ?g pCDI-CMTM1-v17 plasmids separately were 70.8 and 34.7, compared with the control set as 100. When trichostatin A, the inhibitor for histone deacetylase, was used, the repression of androgen receptor could be recovered with trichostatin A treatment,for the relative luciferase activity of the PC3 cells transfected with 1 ?g and 2 ?g pCDI-CMTM1-v17 plasmids and treated with 100 nmol/L trichostatin A rebound to 90.9 and 86.4. Conclusion: CMTM1-v17 is highly expressed in both normal prostate and prostate carcinoma originated cell lines. It may recruit histone deacetylas to inhibit the function of androgen receptor.