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1.
Article in Chinese | WPRIM | ID: wpr-879517

ABSTRACT

OBJECTIVE@#To analyze the results of concurrent hearing and deafness genetic screening and follow up of newborns.@*METHODS@#In total 33 911 babies born to 5 designated hospitals in Nanshan District of Shenzhen city from October 2017 to December 2019 were included. All subjects underwent concurrent hearing and deafness genetic screening covering 21 variants of 4 genes including GJB2, SLC26A4, GJB3 and Mt12SrRNA. For those with positive results, Sanger sequencing was carried out for confirmation.@*RESULTS@#93.32% subjects passed the first-round hearing screening, and 87.01% passed the recheck testing. The overall detection rate was 4.18%. The detection rates for GJB2, SLC26A4, GJB3 and Mt12srRNA variants were 1.98%, 1.58%, 0.37% and 0.25%, respectively. 126 and 84 subjects were found with high risk for delayed-onset and drug-induced hearing loss, respectively. In addition, 4 and 5 subjects were found to harbor homozygous/compound heterozygous variants of the GJB2 and SLC26A4 genes, respectively. Concurrent screening showed that subjects (with heterozygous variants) who did not passed the two round hearing test were as follows: GJB2 with 6.75% in the first round and 2.61% in the second round testing, SLC26A4 (3.3%/1.2%), GJB3 (0.72%/0.14%) and 12SrRNA (0.36%/Nil), respectively. Moreover, the No-pass rate in the subjects with homozygous or compound variants in single gene, heterozygous variant in single gene, heterozygous variant in multiple genes, and homozygous variant in GJB3 gene were significantly higher than the subjects with negative results of genetic screening.@*CONCLUSION@#Concurrent newborn genetic screening can enhance the effectiveness of hearing screening and enable earlier identification and intervention for children with hearing impairment. Follow-up can improve the diagnostic rate for children who are positive for the concurrent screening. Nevertheless, genetic and hearing screening cannot replace the diagnostic testing. It is necessary to conduct comprehensive analysis for the results of genetic and hearing screening and radiological examinations. Sanger sequencing and next-generation sequencing are critical for ascertain the diagnosis.


Subject(s)
China/epidemiology , DNA Mutational Analysis , Deafness/genetics , Follow-Up Studies , Genes/genetics , Genetic Testing/statistics & numerical data , Hearing/genetics , Hearing Tests/statistics & numerical data , Humans , Infant, Newborn , Mutation , Neonatal Screening
2.
Article in Chinese | WPRIM | ID: wpr-828105

ABSTRACT

OBJECTIVE@#To construct a HIV-1 gp120 transgenic mice (gp120 Tg) with vimentin (VIM) gene knockout.@*METHODS@#Female HIV-1 gp120 Tg mice were mated to VIM heterozygote mice (F0). All the offspring mice were derived from these original founders so that both genotypes had the same mixed genetic background. The F1 mice were bred to generate of VIM, VIM, VIM/gp120 Tg and VIM/gp120 Tg mice. PCR was performed for genotyping of the mice, and the expressions of VIM and gp120 in the brain tissues were examined using immunoblotting.@*RESULTS@#The results of PCR showed the presence of the target bands in VIM, VIM, VIM/gp120 Tg and VIM/gp120 Tg mice. In VIM/gp120 Tg mice, gp120 expression was detected throughout the brain regions while no VIM expression was detected.@*CONCLUSIONS@#We generated gp120 transgenic mouse models with VIM gene knockout, which facilitate the exploration of the role of VIM in gp120-induced neurotoxicity.


Subject(s)
Animals , Brain , Disease Models, Animal , Female , HIV Envelope Protein gp120 , HIV-1 , Mice , Mice, Knockout , Mice, Transgenic , Vimentin
3.
Article in Chinese | WPRIM | ID: wpr-801954

ABSTRACT

The research literature of Laportea bulbifera was summarized and analyzed, and its distribution of literature, chemical composition, pharmacological activity, quality control, clinical application and patent approval were summarized, this study can provide reference for the follow-up clinical application and development and utilization of the herb. Taking CNKI and PubMed database as the retrieval platform, keywords and full text as the search items, the Honghema, Honghuoma, Zhuya Aima, Laportea bulbifera (Siebold & Zuccarini) Weddell. and Laportea bulbifera as the search terms, the domestic and foreign paper reports and patent approvals of L. bulbifera from 1989 to 2018 were retrieved. A total of 41 papers and 63 patents were reviewed, the contents of these papers were pharmacological activity, chemical composition and quality control research, the majority of patents were compound. At present, 73 chemical constituents have been isolated and identified from L. bulbifera, and most of them were flavonoids. Flavonoids, catechins and coumarins were selected as the quality control indexes, and most of the pharmacological activities were anti-inflammatory, analgesic and anti-rheumatism. L. bulbifera is highly competitive in the market because of its remarkable pharmacological activities, however, its quality control level in local standard is low and testing items are incomplete. The determination reported in the literature lacks specific indexes to evaluate the quality of L. bulbifera, at the same time, it is necessary to further study its anti-inflammatory mechanism, explore its quality markers and establish the spectrum-effect relationship, so as to effectively control the quality of L. bulbifera and provide documentation for its comprehensive utilization and resource development.

4.
Article in Chinese | WPRIM | ID: wpr-513573

ABSTRACT

Oxidative stress played an important role in the development of diabetes and its complications. Nuclear factor erythroid 2-related factor 2 (NRF2) pathway is one of the most vital endogenous antioxidant pathways. Accumulated evidences indicated that the relationship between diabetes and NRF2 pathway attracted more and more attention in recent years. Our group has devoted ourselves to the researches concerning the chronic complications of diabetes and NRF2 pathways. This review highlighted our recent progresses in the underlying mechanism of the protective role of NRF2 in diabetic nephropathy, diabetic ulcers and diabetic amyotrophy. Finally, the possibility of NRF2 agonists applied to clinical therapy for diabetic chronic complications was explored.

5.
Article in Chinese | WPRIM | ID: wpr-232511

ABSTRACT

<p><b>OBJECTIVE</b>To test the effect of the c-Met inhibitor cabozantinib in inhibiting infections by Listeria monocytogenes (LM) in mice.</p><p><b>METHODS</b>C57BL/6 mice at 6 weeks of age were subjected to intraperitoneal injection of LM and randomized into 4 groups for treatment with intraperitoneal injection of PBS, intragastric administration of cabozantinib (20 µg/g), intraperitoneal injection of ampicillin (Amp, 20 µg/g), or cabozantinib plus Amp. The survival curves were drawn for each group, and the number of bacteria in the blood and brain tissues was determined; serum IL-10 level and NF-κB p65 level in the cerebrospinal fluid (CSF) were assayed, and Evans Blue (EB) content and pathological changes in brain were examined.</p><p><b>RESULTS</b>Compared with PBS-treated mice, the mice treated with cabozantinib showed a significantly higher survival rate, lower bacterial counts in the blood and brain (P<0.05 or 0.001), lower IL-10 (P<0.05) and NF-κB p65 levels (P<0.01), lower brain EB content (P<0.001), and milder pathological changes in the brain. The blood and brain bacterial counts (P<0.001), IL-10 (P<0.01) and NF-κB p65 levels (P<0.001), and brain EB content (P<0.001) were all significantly lower in mice treated with the combination of drugs than in mice treated with cabozantinib alone.</p><p><b>CONCLUSION</b>Cabozantinib can inhibit LM infection in mice and has important values in developing new anti-intracellular infection drug.</p>


Subject(s)
Anilides , Pharmacology , Animals , Brain , Microbiology , Pathology , Injections, Intraperitoneal , Interleukin-10 , Blood , Listeria monocytogenes , Listeriosis , Drug Therapy , Mice , Mice, Inbred C57BL , Pyridines , Pharmacology , Transcription Factor RelA , Cerebrospinal Fluid
6.
Article in Chinese | WPRIM | ID: wpr-286892

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of Lactobacillus rhamnosus GG (LGG) for inhibiting E.coli K1 (E44) adhesion and invasion of an intestinal epithelial cell model with Muc2 gene knockdown established using CRISPR-Cas9 system.</p><p><b>METHODS</b>Two 20-25 bp sgRNAs targeting Muc2 were chemically synthesized to construct CRISPR expression vectors for transfection in wild-type human colonic cancer cell line Ht29. The efficiency of Muc2 knockdown was determined using Western blotting. After assessment of the viability and proliferation of the transfected cells with MTT assay, we evaluated the effects of the probiotics against E44 adhesion and invasion of the cells through a competitive exclusion assay.</p><p><b>RESULTS</b>Transfection of the cells with Lenticrisprv2 plasmid vectors resulted in a cell line with stable Muc2 knockdown by 81%. The inhibitory effects of probiotics against E44 adhesion and invasion of the transfected cells were markedly attenuated, and the relative adhesion and invasion rates of E44 were 72.23% (P<0.05) and 81.49% (P<0.05), respectively.</p><p><b>CONCLUSION</b>Muc2 knockdown causes attenuation of the inhibitory effects of probiotics against E44 adhesion and invasion of the intestinal epithelial cells, suggesting that up-regulation of Muc2 may serve as an important mechanism for the probiotics to reinforce the intestinal barrier and antagonize the pathogenic bacteria, which sheds light on a new strategy for prevention and treatment of bacterial intestinal infections.</p>


Subject(s)
Bacterial Adhesion , CRISPR-Cas Systems , Epithelial Cells , Cell Biology , Microbiology , Escherichia coli , Virulence , Gene Knockdown Techniques , HT29 Cells , Humans , Intestines , Cell Biology , Lactobacillus rhamnosus , Mucin-2 , Genetics , Probiotics , Transfection , Up-Regulation
7.
Article in Chinese | WPRIM | ID: wpr-264066

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of c-Met inhibitor cabozantinib (XL-184) in inhibiting Listeria monocytogenes (LM) from invading Caco-2 cells to reduce the cell injury.</p><p><b>METHODS</b>The cell invasion capacity of LM was assayed in Caco-2 cells incubated with different doses of XL-184 for different durations. Caco-2 cells incubated with XL-184 were seeded on the upper room of the transwell chamber, and the cell monolayer was exposed to LM infection followed by addition of horseradish peroxidase (HRP). The trans-epithelial electric resistance (TEER), HRP concentration and LM colony-forming unit (CFU) were measured in the cell monolayer. Fluorescent staining was used to evaluate the cell viability, and LDH release from the cells was examined to assess the changes in cell membrane permeability.</p><p><b>RESULTS</b>XL-184 significantly decreased LM invasion rate in Caco-2 cells in a dose- and time-dependent manner (P=0.000), and this effect was enhanced by co-incubation of the cells with ampicillin (P<0.05). In the cell membrane permeability assay in the monolayer cells, XL-184 markedly inhibited LM-induced reduction of TEER (P<0.05) and significantly suppressed LM-induced enhancement of cell membrane permeability shown by reduced HRP concentration and LM count in the lower chamber (P=0.000). The cells infected with LM showed significantly lowered cell viability, which was rescued by XL-184 (P<0.01); XL-184 also dose-dependently reduced LDH release from the cells (P<0.05).</p><p><b>CONCLUSIONS</b>XL-184 can suppress LM invasion in Caco-2 cells to reduce the cell injury, suggesting its value as a promising candidate agent for prevention and treatment of LM infections.</p>


Subject(s)
Anilides , Pharmacology , Caco-2 Cells , Cell Membrane Permeability , Cell Survival , Humans , Listeria monocytogenes , Pyridines , Pharmacology
8.
Article in Chinese | WPRIM | ID: wpr-491767

ABSTRACT

Objective To use mouse astrocytes elevated gene‐1 (AEG‐1) monoclonal fluorescent antibody for detecting tumor cells in malignant serous cavity effusions .Methods The expression of AEG‐1 in serous cavity effusion exfoliated cells by PCR and Western‐blot ;the mouse monoclonal anti‐AEG‐1 fluorescent antibody and tumor cells in malignant serous cavity effusions were co ‐incubated ,meanwhile ,the serous cavity effusions in benign lesions were taken as the negative control .Results The AEG‐1 expres‐sion was positive in malignant serous cavity effusions exfoliated cells ,while which in benign lesion serous cavity effusion was nega‐tive or weakly positive ;meanwhile the results of direct labelling in mouse anti AEG‐1 monoclonal antibodies were consistent with the results by PCR and Western‐blot .Conclusion Mouse anti AEG‐1 monoclonal fluorescence antibody can provide certain theoret‐ical basis for the detection of tumor cells in serous cavity effusion .

9.
China Pharmacy ; (12): 4988-4989,4990, 2016.
Article in Chinese | WPRIM | ID: wpr-605874

ABSTRACT

OBJECTIVE:To observe the clinical efficacy and safety of piperazine ferulate combined with glutathione in the treatment of diabetic nephropathy. METHODS:80 patients with diabetic nephropathy in our hospital were divided into observation group and control group according to random number table,with 40 cases in each group. Both groups was given general treatment as blood glucose,blood lipid and blood pressure control. Control group was additionally given Reduced glutathione tablets 400 mg, tid,on the basis of general treatment. Observation group was additionally given Piperazine ferulate tablets 100 mg,tid,on the ba-sis of control group. Both groups were treated for 12 weeks. The fasting plasma glucose(FPG),2 h postprandial plasma glucose(2 hPG),blood pressure,blood lipid,serum creatinine (Scr),blood urea nitrogen (BUN),24 h urinary total protein and albumin, urine β2 microglobulin(β2-MG)and N-acetyl-β-glucosaminidase(NAG)of 2 groups were detected before and after treatment. The occurrence of ADR was observed. RESULTS:Before treatment,there was no statistical significance in FPG,2 hPG,blood pres-sure,blood lipid,Scr,BUN,24 h urinary total protein and albumin,urine β2-MG and NAG between 2 groups(P>0.05). After treatment,above indexes of 2 groups were improved significantly,and the observation group was better than the control group, with statistical significance(P<0.05). No obvious ADR was found in 2 groups. CONCLUSIONS:In the treatment of diabetic ne-phropathy,piperazine ferulate combined with glutathione can improve blood glucose,blood pressure,blood lipid levels and renal function with good safety.

10.
Article in Chinese | WPRIM | ID: wpr-239154

ABSTRACT

<p><b>OBJECTIVE</b>To explore the role of CD44 in monocyte adhesion to human brain microvascular endothelial cells (HBMECs) and monocyte migration across an in vitro model of blood-brain barrier (BBB) infected by Cryptococcus neoformans (Cn).</p><p><b>METHODS</b>An in vitro blood-brain barrier model was constructed using a transwell chamber covered with a HBMEC monolayer. The wild-type strain of Cn B4500FO2, TYCC645#32 strain with CPS1 gene deletion and PCIP strain with CPS1 complementation were chosen to infect the monolayer HBMECs. THP-1 cells were added to the upper chamber of transwell, and the relative migration rate was determined by counting the number of the cells entering the lower chambers. The inhibitory effects of anti-CD44 monoclonal antibody and the CD44 inhibitor bikunin were examined on THP-1 binding to and migration across HBMECs.</p><p><b>RESULTS</b>Cn infection of the HBMECs caused markedly enhanced THP-1 cell adhesion and migration across the monolyers (P<0.01) dependent on Cn concentration and exposure time. Addition of anti-CD44 monoclonal antibody and bikunin significantly lowered THP-1 adhesion and migration rates in the BBB model with Cn-infected HBMECs (P<0.01) with a dose dependence of the antibody (within 0-1 µg) and inhibitor (within 0-20 nmol/L). Both THP-1 adhesion rate and migration rate were lowered in the BBB model infected with CPS1 gene-deleted Cn but increased in the model infected with the complemented strain compared with those in the wild-type strain-infected model.</p><p><b>CONCLUSION</b>In the in vitro BBB model, CD44 expressed on HBMECs may play an essential role in monocyte adhesion to and migration across the BBB. The capsular hyaluronic acid may mediate Cn-induced monocyte adhesion and migration.</p>


Subject(s)
Blood-Brain Barrier , Allergy and Immunology , Microbiology , Brain , Cell Biology , Microbiology , Cell Line , Cryptococcosis , Allergy and Immunology , Cryptococcus neoformans , Endothelial Cells , Microbiology , Humans , Hyaluronan Receptors , Metabolism , Monocytes , Cell Biology
11.
Article in Chinese | WPRIM | ID: wpr-481746

ABSTRACT

BACKGROUND:There is a close connection between the occurrence and development of tumor stem cels and ovarian cancer. CD90+ is an important tumor stem cel marker. OBJECTIVE: To explore the biological characteristics of CD90+ tumor stem cels in ovarian cancer cels. METHODS:The CD133 and CD90 positive rate of SKOV3 and primary ovarian cancer cels were detected by flow cytometry. The CD90+ and CD90- relative expression in stem cels and epithelium was detected by RT-PCR. Transwel invasion assay was employed to observe the cel invasion ability, clone formation test was done to observe cel proliferation and differentiation capacity, suspension bal test was adopted to observe pluripotent stem cels. The tumor formation time and tumor formation rate were observed by limited tumor dilution in immunodeficient mice. RESULTS AND CONCLUSION:The positive rates of CD133 and CD90 in SKOV3 were significantly lower than those in primary ovarian cancer cels. The expression of CD133 and OCT4 in CD90+cels of SKOVS was significantly higher than that in CD90-cels of SKOVS. The expression of CD44, CD133, acetaldehyde dehydrogenase-1 and OCT4 in CD90+stem cels of primary ovarian cancer cels was significantly higher than that in CD90-stem cels of primary ovarian cancer cels. There were significant differences in the epithelial-mesenchymal related gene expressions between CD90-and CD90+stem cels of SKOV3 and primary ovarian cancer cels. With the increase of inoculated cels, the tumor formation rate of CD90-and CD90+ cels was increased continuously, but the tumor formation time was decreased. The tumor rate of CD90-cels was lower than that of CD90+cels. The number of transmembrane cels, cel clones and suspended cel bals was significantly higher in the CD90+ stem cels than the CD90-stem cels. These findings indicate that in ovarian cancer cels, CD90+stem cels can highly express stem cel-related genes and epithelial-mesenchymal related genes, which have a higher invasion, proliferation and differentiation ability, as wel as tumorigenic and pluripotent ability.

12.
Article in Chinese | WPRIM | ID: wpr-600134

ABSTRACT

Objective To construct anti-astrocyte elevated gene-1(AEG-1)single-chain variable antibody (V23)prokaryotic ex-pression vector,and to conduct the protein purification and immunological activity detection.Methods The Primer5 software was applied to design the primers aiming at the gene sequence of the antibody anti-AEG-1 single-chain variable region for constructing the prokaryotic expression plasmid of PRsetC/V23.After the enzyme digestion by the restriction enzyme Pst1 and correct DNA se-quencing,the prokaryotic expression plasmid was led to E.coli BL21 ,the prokaryotic expression engineering strain containing the V23 gene was constructed.After the induction with IPTG,the interest protein was purified by the magnetic beads with the HIS tag,and the content of interest protein was determined by the SDS-PAGE electrophoresis.Western blot and ELISA were adopted to detect the immune activity of the nti-AEG-1 single-chain variable region antibody.Results For the constructed prokaryotic expres-sion plasmid PRsetC/V23,the single enzyme digestion and sequencing analysis displayed that the constructed V23 gene was com-pletely consistent to the designing sequences.After IPTG induction,SDS-PAGE electrophoresis showed an apparent protein band at 31×103 ,the Western blot detection showed a specific AEG-1 response band at 80 ×103 ,the ELISA test showed the positive re-sults.Conclusion The PRsetC/V23 prokaryotic expression plasmid and the V23 prokaryotic expression engineering strain are suc-cessfully constructed,this engineering strain can express anti-AEG-1 single-chain variable region antibody protein,and the protein has good immune activity.

13.
Chongqing Medicine ; (36): 4460-4461, 2014.
Article in Chinese | WPRIM | ID: wpr-458327

ABSTRACT

Objective To detect serum galectin 3 levels in patients with chronic kidney disease (CKD) and explore its clinical significance .Methods The galectin 3 levels the serum of in 38 CKD patients and 34 healthy controls were determined by ELISA . All kidney function was measured by automatic biochemical analyzer .The relation between serum galectin 3 levels and the level of kidney function was analyzed by use of t test .Results The serum galectin 3 levels in CKD and healthy controls groups were (1 .22 ± 1 .01)ng/mL and (3 .03 ± 2 .06)ng/mL ,P<0 .05 .There was close negative correlation between serum levels of galectin 3 and Cr ,CysC(P<0 .05) .Conclusion Serum galectin 3 of CKD patients reduces significantly and correlates with kidney function . Detecting on s galectin 3 is helpful for chronic kidney disease diagnosis and therapeutic effect evaluation .

14.
Article in Chinese | WPRIM | ID: wpr-456520

ABSTRACT

To investigate the plasma thioredoxin-interacting protein ( TXNIP ) levels in different glucose tolerance groups and discuss the relationship between TXNIP and insulin resistance/β-cell dysfunction in diabetes and prediabetes, and to investigate the potential relationship between TXNIP and interleukin-1β( IL-1β) . According to oral glucose tolerance test, 93 participants were divided into 3 groups:diabetes mellitus group, prediabetes group, and normal glucose tolerance group. Plasma TXNIP, IL-1β, and other biochemical indices were measured. The relationship between TXNIP and glucose, IL-1β, homeostasis model assessment for insulin resistance ( HOMA-IR) , and homeostasis model assessment forβcell function ( HOMA-β) were analyzed by using multiple linear regression techniques and Pearson’s linear correlation analysis. Plasma TXNIP level was higher in prediabetes group compared with normal glucose tolerance group, but lower in prediabetes group compared with diabetes mellitus group[(355. 35±31. 88 vs 274. 36±33. 86, 426. 16±63. 15)pg/ml, P<0. 01 or P<0. 05]. TXNIP was positively correlated with IL-1βand HOMA-IR, but negatively correlated with HOMA-β. Multiple linear regression analysis indicated that IL-1βexerted significant influence on TXNIP ( P<0. 05 ). Plasma TXNIP level is affected by blood glucose concentration. There is a close relationship between TXNIP and IL-1β. In prediabetes patient, the TXNIP levels have already been raised.

15.
Article in Chinese | WPRIM | ID: wpr-356960

ABSTRACT

<p><b>OBJECTIVE</b>To study the role of outer membrane protein T (OmpT) in the pathogenesis of uropathogenic Escherichia.coli.</p><p><b>METHODS</b>In cultured human bladder epithelial cell line 5637, we examined the adhesion ability of wild-type (CFT073), ompT gene knockout (COTD), and revertant (pST) strains of E.coli to the cells and the extracellular matrix (ECM). The expressions of the adhesion gene iha and virulence gene iroN were detected by real-time PCR. Murine models of urinary tract infection with the 3 strains were established to evaluate the bacterial burden of the bladder and kidney tissue and bacterial counts in blood. We also detected the expressions of interleukin-6 (IL-6) and IL-8 in the bladder and kidney tissues of the mice.</p><p><b>RESULT</b>The COTD strain showed a significantly lower cell adhesion rate than CFT073 strain [(4.62∓0.39)% vs (8.81∓1.13)%, P<0.05] with also a lower ECM-adhesion rate [(4.95∓0.59)% vs (8.85∓0.79)%, P<0.05]. The mRNA expressions of iha and iroN in CFT073 strain were 2.1 and 3.8 times that of COTD strain. In the mouse model, the mean bacterial load of CFT073 strain in the bladder tissue was 6.36∓0.06, significantly greater than that of COTD (6.01∓0.07) and revertant (6.29∓0.06) strains (P<0.05); the bacterial load of CFT073 strain in the kidney tissue was also significantly higher than that of COTD strain (6.25∓0.05 vs 5.87∓0.06, P<0.05). In mice infected with the wild-type, knockout, and revertant strains, the detection rates of IL-6, which were identical to those of IL-8, in the inflammatory bladder and kidney tissues were 60%, 12.5%, and 50%, respectively.</p><p><b>CONCLUSIONS</b>OmpT may regulate the expression of the adhesion gene iha and the transferrin gene iroN to affect the adhesion of uropathogenic E.coli to host cells.</p>


Subject(s)
Animals , Bacterial Adhesion , Bacterial Load , Bacterial Outer Membrane Proteins , Metabolism , Cell Line, Tumor , Escherichia coli Infections , Pathology , Escherichia coli Proteins , Metabolism , Gene Knockout Techniques , Humans , Inflammation , Interleukin-6 , Metabolism , Interleukin-8 , Metabolism , Kidney , Microbiology , Mice , Peptide Hydrolases , Metabolism , Receptors, Cell Surface , Metabolism , Urinary Bladder , Microbiology , Urinary Tract Infections , Microbiology , Pathology , Uropathogenic Escherichia coli , Virulence
16.
Article in Chinese | WPRIM | ID: wpr-322082

ABSTRACT

<p><b>OBJECTIVE</b>To investigate MUC2 expression in rat colons induced by probiotics and its effects on the inhibition of E.coli K1 (E44) penetration of the intestinal barrier by probiotics.</p><p><b>METHODS</b>SD rats were subjected to intragastric administration of probiotics, E44, or probiotics +E44 on a daily basis for 7 days, and MUC2 expression in the colons was determined by RT-PCR. MUC2-targeted shRNA (shRNA MUC2) and scrambled shRNA plasmids (shRNA NC) were respectively transfected into Lovo cells, and the efficiency of MUC2 knockdown was determined using qRT-PCR. Competitive exclusion assay was used to evaluate the effects of the probiotics against E44 adhesion and invasion.</p><p><b>RESULTS</b>Intestinal MUC2 mRNA expression was up-regulated in the rats after intragastric administration of probiotics, while E44 administration caused significantly lowered MUC2 expression. MUC2 expression was down-regulated (by 66.7%) by transfection with shRNA MUC2 in Lovo cells as compared with the negative control and mock control cells. The inhibition of E44 adherence and invasion by probiotics was significantly attenuated in transfected Lovo cell culture (in which the relative adhesion and invasion rates of E44 were 56.64% and 66.64%, respectively) as compared with those in the control group.</p><p><b>CONCLUSION</b>The up-regulation of MUC2 in rat colons can be one of the mechanisms of the probiotics in antagonizing the translocation of the pathogenic bacteria. Silencing MUC2 expression causes attenuated inhibitory effect of the probiotics on E. coli K1 penetration across human intestinal epithelial cells.</p>


Subject(s)
Animals , Animals, Newborn , Cell Line, Tumor , Colon , Metabolism , Microbiology , Escherichia coli , Virulence , Escherichia coli Infections , Genetics , Female , Gene Expression Regulation, Neoplastic , Gene Silencing , Humans , Mucin-2 , Genetics , Probiotics , Pharmacology , RNA, Messenger , Genetics , RNA, Small Interfering , Rats , Rats, Sprague-Dawley , Transfection
17.
Article in Chinese | WPRIM | ID: wpr-265670

ABSTRACT

<p><b>OBJECTIVE</b>To explore the feasibility of inducing fungal infection by Cryptococcus neoformans aerosol inhalation in immunosuppressed Balb/c mice.</p><p><b>METHODS</b>Twenty-four Balb/c mice were randomized into cyclophosphamide (CTX) group and control group (n=12). The mice in CTX group were subject to inhalation of Cryptococcus neoformans aerosol prepared using a ultrasonic nebulizer 4 days after intraperitoneal CTX injection, and the control mice received the inhalation only. The leukocyte count and changes in appetite, body weight, and behaviors of the mice were observed after the treatments. On days 3 and 7 after the inoculation, the mice were sacrificed to analyze the fungal counts in brain and lung tissue homogenates and examine the pulmonary pathologies.</p><p><b>RESULTS</b>CTX injection caused lowered appetite and body weight loss in the mice, and significantly reduced the leukocyte counts (2.77∓0.45 vs 8.26∓0.56, P<0.05). At days 3 and 7 after inoculation, the Cryptococci load in the lungs increased obviously in CTX group with a colony-forming unit (CFU) of the lungs of 271.67∓122.22 and 41.67∓0.28, respectively, significantly higher than that in the control group (60.00∓43.36 and 3.00∓5.30, respectively, P<0.05). In CTX group, the CFU was 10.17∓5.42 and 9.17∓6.34 in the peripheral blood and 6.83∓4.92 and 11.00∓5.44 in the brain tissue at days 3 and 7, respectively, whereas no Cryptococci was detected in the peripheral blood or in the brain tissue in the control group. Pathological examination of the lungs revealed destruction of normal alveolar structure in CTX group, with numerous infiltrating inflammatory cells and visible yeast.</p><p><b>CONCLUSION</b>Inhalation of Cryptococcus neoformans aerosol can cause fungal infection in the lungs of immunosuppressed Balb/c mice.</p>


Subject(s)
Aerosols , Animals , Brain , Microbiology , Colony Count, Microbial , Cryptococcosis , Microbiology , Cryptococcus neoformans , Disease Models, Animal , Immunocompromised Host , Inhalation , Lung Diseases, Fungal , Microbiology , Male , Mice , Mice, Inbred BALB C , Ultrasonics
18.
Article in Chinese | WPRIM | ID: wpr-268956

ABSTRACT

<p><b>OBJECTIVE</b>To explore the role of ompT gene in uropathogenic E. coli (UPEC) CFT073 strain in urinary tract infection (UTI).</p><p><b>METHODS</b>An ompT deletion mutant (COTD) was generated by λ Red recombineering in the UPEC CFT073 strain, which was characterized by PCR and sequencing. C57B/L6 mouse models of acute UTI with the mutant and wild-type strains were established to compare the colonization abilities of the two strains in the bladder. The adhesion of CFT073 mutant to human unthelial 5637 cells was also investigated in vitro.</p><p><b>RESULTS</b>PCR and DNA sequencing confirmed the loss of ompT gene in the mutant COTD. The in vitro adhesion rate of the mutant strain COTD to 5637 cells was (6.7±2.2)%, significantly lower than that of (8.3±1.9)% of the wild-type strain (P<0.05). In the murine models of acute UTI, the mutant strain showed a mean colonization number of about (17±8)×10⁴ cfu, which was significantly lower than that of (7∓2)×10⁵ cfu of the wide-type CFT073 strain (P<0.05).</p><p><b>CONCLUSION</b>OmpT gene can be involved in the colonization of UPEC in the bladder tissue and plays an important role in the pathogenesis of UPEC-induced UTI.</p>


Subject(s)
Animals , Bacterial Proteins , Genetics , Cell Line , Escherichia coli Infections , Microbiology , Escherichia coli Proteins , Genetics , Gene Knockout Techniques , Humans , Mice , Mice, Inbred C57BL , Porins , Genetics , Urinary Tract Infections , Microbiology , Uropathogenic Escherichia coli , Genetics
19.
Article in Chinese | WPRIM | ID: wpr-307906

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of HIV-1 gp41 ectodomain (gp41-I90) on the cytoskeletal changes in human brain microvascular endothelial cells (HBMECs) induced by Cryptococcus neoformans.</p><p><b>METHODS</b>HBMECs were cultured on collagen-coated chamber slide or transwell to allow the formation of cell monolayers. After pre-treatment with gp41-I90 and infection with Cryptococcus neoformans, the HBMECs were examined for the expression of actin or filamin by immunofluorescence assay. HRP permeability of the HBMECs treated with gp41-I90 was detected by ELISA. Transcytosis of Cryptococcus neoformans through the gp41-I90-treated HBMECs was detected by direct counting from a hemocytometer.</p><p><b>RESULTS</b>gp41-I90 obviously enhanced the cytoskeletal changes of the HBMECs infected by Cryptococcus neoformans, causing curved and sparse filamentous arrangement of actin and filamin. gp41-I90 treatment also resulted in obviously increased HRP permeability of the cells and transcytosis of Cryptococcus neoformans.</p><p><b>CONCLUSION</b>gp41- I90 enhances Cryptococcus neoformans binding to HBMECs, which is related to its effect in enhancing Cryptococcus neoformans-induced cytoskeletal changes of the cells.</p>


Subject(s)
Brain , Cells, Cultured , Cryptococcosis , Pathology , Cryptococcus neoformans , Virulence , Cytoskeleton , Metabolism , Endothelial Cells , Cell Biology , Microbiology , Endothelium, Vascular , Cell Biology , Metabolism , HIV Envelope Protein gp41 , Pharmacology , Humans , Microcirculation
20.
Article in Chinese | WPRIM | ID: wpr-416913

ABSTRACT

By using the Medical Case Inquiry System and registries of infant's birth,the number of women with gestational diabetes mellitus(GDM)and the total number of women delivering in the First and Second Affiliated Hospital of Chongqing Medical University,Chongqing Xinqiao Hospital,and Chongqing Health Center For Women and Children from 2005 to 2009 were obtained.The data of 540 pregnant women with GDM were further analyzed.From 2005 to 2009,the incidence of GDM increased from 2.29% to 3.81 %(P0.05).From 2005 to 2009,the other related factors,including the average maternal age,the constituent ratio of women with advanced maternal age,pregnancy history,delivery history,and family history of diabetes showed insignificant changes(P>0.05).

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