The gene encoding fibronectin-binding protein A(FnBA) was amplified from Staphylococcus aureus chromosomal DNA by PCR.Using the T-A cloning technique,the PCR product about 1 735 bp in length was cloned into a pMD18-T vector and was designated plasmid pMD18-FnBA.pMD18-FnBA and pET28a(+) were digested by BamHⅠ and HindⅢ,then the purified FnBA gene was sub-cloned into expression vector pET28a(+),and the prokaryotic expression vector pET28a-FnBA was constructed.The constructed pET28a-FnBA was transformed into E.coli BL21 competent cells and then induced by IPTG(1 mmol/L).SDS-PAGE analysis revealed a band of approxi-mately 85 000 in molecular weight from the induced E.coli BL21 competent cells.Western-blotting analysis indicated that the protein had antigenic activity of FnBA.
Objective:To compare the influences of retrorsine on hepatocytes proliferation in mice and rats after liver injury,so as to investigate the feasibility of using retrorsine for establishment of liver cell transplantation model in mice.Methods: Male mice and rats were pretreated with 2 injections of retrorsine(70 mg/kg for mice and 30 mg/kg for rats)(as retrorsine-treated group,n=30) at 2 weeks interval or saline(as nontreated group,n=30).A single injection of carbon tetrachloride(CCl_(4), 0.5 mg/kg) was given to all animals 4 weeks after the final injection of retrorsine or saline.At 0(before administration),1,2,3,4,6,15 d after CCl_(4) administration,the animals were sacrificed and their livers were subjected to H-E staining and Ki-67 antibody immunohistochemistry analysis to evaluate the pathological changes and hepatocyte proliferation.Results: The liver in rats treated with retrorsine displayed obvious megalocytosis,small bile duct hyperplasia,and small hepatocyte hyperplasia(forming nodule);no such changes were found in non-treated group.However,the liver in mice treated with retrorsine displayed hepatocyte degeneration and necrosis in the perivenous areas and the same was true to the liver in non-treated mice.Ki-67 immunohistochemistry analysis showed that in rats treated with retrorsine,the positive hepatocytes,mainly found in small hepatocyte nodules,were obviously less than those in control group 3 d after CCl_(4)administration(P0.05).Ki-67 positive hepatocytes in mice treated with retrorsine were abundant and almost more than those in control group at all time points,especially 4 d after CCl_(4) administration(P0.01),with the same changing tendency in both groups.Conclusion: Retrorsine can obviously inhibit hepatocyte proliferation after liver injury and is suitable for liver cell transplantation in rats,while it is the contrary in mice.
Lipid oxidation can produce lipid oxidation products (LOPs), which further react with proteins and affect their structure and digestibility, although the underlying mechanism remains unclear. Herein, we investigated the conformation and digestibility of proteins induced by LOPs after thermal treatment. Digestibility of myoglobin (Mb) affected by trans,trans,-2,4-decadienal (2,4-Dec) was significantly reduced under high temperature (100-180 °C). The peptides digested from Mb modified with 2,4-Dec during thermal processing revealed that the quantity of peptides decreased with increasing 2,4-Dec concentrations. Proteomic analysis showed that 2,4-Dec covalently binds to Mb, and the extent of modification was in the following order: lysine > histidine > arginine. Moreover, the secondary structure, intrinsic fluorescence, and surface hydrophobicity results suggested that 2,4-Dec induced changes in Mb, leading to a tighter spatial structure and aggregation, and exposure of fewer recognition sites of the enzyme and thermal treatment assisted these changes in the structure. Meanwhile, molecular dynamics simulations elucidated the molecular mechanisms underlying the effect of 2,4-Dec and temperature on the digestion and structure of Mb.
Hyperuricemia nephropathy (HN) is a metabolic disease characterized by tubular damage, tubulointerstitial fibrosis, and uric acid kidney stones and has been demonstrated to be associated with hyperuricemia. Coffee leaf tea is drunk as a functional beverage. However, its prevention effects on HN remain to be explored. This study showed that coffee leaf tea extracts (TE) contain 19 polyphenols, with a total content of 550.15 ± 27.58 mg GAE/g. TE decreased serum uric acid levels via inhibiting XOD activities and modulating the expression of urate transporters (GLUT9, OAT3, and ABCG2) in HN rats. TE prevented HN-induced liver and kidney damage and attenuated renal fibrosis. Moreover, it upregulated the abundance of SCFA-producing bacteria (Phascolarctobacterium, Alloprevotella, and Butyricicoccus) in the gut and reversed the amino acid-related metabolism disorder caused by HN. TE also decreased the circulating LPS and d-lactate levels and increased the fecal SCFA levels. This study supported the preliminary and indicative effect of coffee leaf tea in the prevention of hyperuricemia and HN.
Background Urbanization has an ecological and evolutionary effect on urban microorganisms. Microorganisms are fundamental to ecosystem functions, such as global biogeochemical cycles, biodegradation and biotransformation of pollutants, and restoration and maintenance of ecosystems. Changes in microbial communities can disrupt these essential processes, leading to imbalances within ecosystems. Studying the impact of human activities on urban microbes is critical to protecting the environment, human health, and overall urban sustainability. Methods In this study, bacterial communities in the sediments of an urban artificial river were profiled by sequencing the 16S rRNA V3-V4 region. The samples collected from the eastern side of the Jiusha River were designated as the JHE group and were marked by persistent urban sewage discharges. The samples collected on the western side of the Jiusha River were categorized as the JHW group for comparative analysis. Results The calculated alpha diversity indices indicated that the bacterial community in the JHW group exhibited greater species diversity and evenness than that of the JHE group. Proteobacteria was the most dominant phylum between the two groups, followed by Bacteroidota. The relative abundance of Proteobacteria and Bacteroidota accumulated in the JHE group was higher than in the JHW group. Therefore, the estimated biomarkers in the JHE group were divided evenly between Proteobacteria and Bacteroidota, whereas the biomarkers in the JHW group mainly belonged to Proteobacteria. The Sulfuricurvum , MND1 , and Thiobacillus genus were the major contributors to differences between the two groups. In contrast to JHW, JHE exhibited higher enzyme abundances related to hydrolases, oxidoreductases, and transferases, along with a prevalence of pathways associated with carbohydrate, energy, and amino acid metabolisms. Our study highlights the impact of human-induced water pollution on microorganisms in urban environments.
Objective
To investigate the chest imaging features in H7N9 subtype human avian influenza.
Methods
The clinical and imaging data of 7 patients with H7N9 subtype avian influenza were collected and the chest imaging manifestations were analyzed.
Results
The main clinical symptoms of H7N9 patients were fever (n=7) and cough (n=4).The ground-glass opacities or consolidations in pulmonary segmental or lobar distribution were showed (n=5) at the initial stage (within 3 days).Air bronchogram could be found in 4 cases.The bilateral lungs were rapidly involved with multiple or diffuse distribution (n=7) at the progressive stage (3—6 days),and 4 patients were died at this stage.The linear,reticular and honeycomb shadows were seen with partial absorption of lesions (n=2).
Conclusion
The main imaging features of H7N9 subtype human avian influenza are the segmental or lobar consolidations on the initial stage and rapid involvement of bilateral lungs with disease progression.
Key words:
Influenza in birds; Pneumonia; Radiography; Tomography,X-ray computed
[Objective] To know human immunodeficiency virus(HIV)infection situation in tuberculosis patients and the clinical characters of HIV infected patients complicated with tuberculosis.Methods The anti-HIV of every new tuberculosis patients in urban districts of Chengdu City was tested by ELISA method first.There were 3 447 new patients tested from 2006-04-01 to 2007-03-31.If the result was positive,the patient must be determined by confirm test.The clinical characters of patients diagnosed as HIV infection and tuberculosis were analyzed.Results 23 cases’anti-HIV were positive in all 3 447 cases,and positive rate was 6.7‰.There were 20 male cases and 3 female cases.The average age was 37.4 years.There were 17 cases of migrant unemployed men and migrant workers(73.9%),2 cases of migrant businessmen(8.7%),and 4 cases with registered permanent residence in Chengdu City(17.4%).In 23 patients with HIV infection,there were 12 patients(52.2%)were diagnosed as pulmonary tuberculosis,5 patients(21.7%)pulmonary tuberculosis and extra-pulmonary tuberculosis,2 patients(8.7%)tuberculous pleurisy,and 4 patients(17.4%) extra-pulmonary tuberculosis.In 23 patients,there were 13 patients(56.5%)were diagnosed as other opportunity infection except tuberculosis and the infection rate was significantly higher than that of in-patients in the same period(P﹤0.01).The positive rate of PPD test was 9.5%,and the rate was significantly lower than that of in-patients in the same period(P﹤0.01).In 17 patients who were diagnosed as HIV infection and pulmonary tuberculosis,2 patients were smear-positive,and the rate of smear-positive was significantly lower than that of in-patients in the same period(P﹤0.05).In the 17 patients,14 patients(82.4%)had main non-typical X-ray and chest CT features,and the rate of non-typical features was significantly higher than that of in-patients in the same period(P﹤0.01).Conclusion Symptoms of patients diagnosed as HIV infection and tuberculosis are not characteristic.The positive rate of PPD test is low.The patients complicated with opportunity infection are common.Patients diagnosed as HIV infection and pulmonary tuberculosis have non-typical X-ray features and low smear-positive rate.Consequently making a wrong diagnosis is common.Physician should be alert the occurrence of HIV infection and tuberculosis.
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