Application of copy number variation sequencing in patients with intellectual disability/developmental delay and autistic spectrum disorder / 中华医学遗传学杂志

OBJECTIVE@#To assess the value of copy number variation sequencing (CNV-seq) for the diagnosis of children with intellectual disability (ID), developmental delay (DD), and autistic spectrum disorder (ASD).@*METHODS@#Forty patients with ID/DD/ASD referred to Nanshan Maternity and Child Health Care Hospital from September 2018 to January 2022 were enrolled. G-banded karyotyping analysis was carried out for the patients. Genomic DNA was extracted from peripheral blood samples and subjected to CNV-Seq analysis to detect chromosome copy number variations (CNVs) in such patients. ClinVar, DECIPHER, OMIM and other database were searched for data annotation.@*RESULTS@#Among the 40 patients (including 30 males and 10 females), 16, 15 and 6 were diagnosed with ID, DD and ASD, respectively. One patient had combined symptoms of ID and DD, whilst the remaining two had combined ID and ASD. Four patients were found with abnormal karyotypes, including 47,XY,+mar, 46,XY,inv(8)(p11.2q21.2), 46,XX,del(5)(p14) and 46,XX[76]/46,X,dup(X)(p21.1q12). Chromosome polymorphism was also found in two other patients. CNV-seq analysis has detected 32 CNVs in 20 patients (50.0%, 20/40). Pathogenic CNVs were found in 10 patients (25.0%), 15 CNVs of uncertain clinical significance were found in 12 patients (30.0%), and 7 likely benign CNVs were found in 4 patients (10.0%).@*CONCLUSION@#Chromosome CNVs play an important role in the pathogenesis of ID/DD/ASD. CNV-seq can detect chromosomal abnormalities including microdeletions and microduplications, which could provide a powerful tool for revealing the genetic etiology of ID/DD/ASD patients.

Plasma microfibrillar associated protein 5 level in patients with polycystic ovary syndrome / 中华内分泌代谢杂志

Objective:To assess plasma microfibrillar associated protein 5(MFAP5) level in patients with polycystic ovary syndrome(PCOS), and to explore its relationship with glucose and lipid metabolism as well as sex hormones.Methods:Fifty PCOS patients and 65 healthy female subjects were selected as PCOS group and control group, respectively. Clinical data and plasma MFAP5 levels between the two groups were compared.Results:The plasma MFAP5 level in PCOS group was significantly higher than that in control group( P<0.01), and the plasma MFAP5 level in PCOS overweight subgroup was higher than that in control subgroup( P<0.01). No difference was observed in plasma MFAP5 level between the two non-overweight subgroups( P>0.05). Correlation analysis showed that plasma MFAP5 level was positively correlated with waist circumference, low density lipoprotein-cholesterol, fasting insulin, homoeostasis model assessment of insulin resistance index(HOMA-IR), HbA 1C, testosterone, LH/FSH ratio, and leukocyte( P<0.05 or P<0.01). There was no significant correlation of MFAP5 with body weight, body mass index(BMI), hip circumference, waist hip ratio, high density lipoprotein-cholesterol(HDL-C), triglyceride, total cholesterol, and blood glucose( P>0.05). In PCOS group, plasma MFAP5 level was positively correlated with body weight, BMI, waist circumference, hip circumference, total cholesterol, and leukocyte( P<0.05 or P<0.01). There was no significant correlation of MFAP5 with waist hip ratio, HDL-C, triglyceride, blood glucose, fasting insulin, HOMA-IR, leukocyte, and sex hormones( P>0.05). Multivariate logistic regression analysis showed that MFAP5 was an independent risk factor for PCOS( P<0.05). Conclusion:Plasma MFAP5 level is increased in PCOS patients and is closely related to BMI, waist circumference, hip circumference, and total cholesterol. Plasma MFAP5 is an independent risk factor for PCOS, which may be involved in the pathogenesis of PCOS.

Identification of Key Genes and Pathways in Peripheral Blood Mononuclear Cells of Type 1 Diabetes Mellitus by Integrated Bioinformatics Analysis

Background@#The onset and progression of type 1 diabetes mellitus (T1DM) is closely related to autoimmunity. Effective monitoring of the immune system and developing targeted therapies are frontier fields in T1DM treatment. Currently, the most available tissue that reflects the immune system is peripheral blood mononuclear cells (PBMCs). Thus, the aim of this study was to identify key PBMC biomarkers of T1DM. @*Methods@#Common differentially expressed genes (DEGs) were screened from the Gene Expression Omnibus (GEO) datasets GSE9006, GSE72377, and GSE55098, and PBMC mRNA expression in T1DM patients was compared with that in healthy participants by GEO2R. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and protein-protein interaction (PPI) network analyses of DEGs were performed using the Cytoscape, DAVID, and STRING databases. The vital hub genes were validated by reverse transcription-polymerase chain reaction using clinical samples. The disease-gene-drug interaction network was built using the Comparative Toxicogenomics Database (CTD) and Drug Gene Interaction Database (DGIdb). @*Results@#We found that various biological functions or pathways related to the immune system and glucose metabolism changed in PBMCs from T1DM patients. In the PPI network, the DEGs of module 1 were significantly enriched in processes including inflammatory and immune responses and in pathways of proteoglycans in cancer. Moreover, we focused on four vital hub genes, namely, chitinase-3-like protein 1 (CHI3L1), C-X-C motif chemokine ligand 1 (CXCL1), matrix metallopeptidase 9 (MMP9), and granzyme B (GZMB), and confirmed them in clinical PBMC samples. Furthermore, the disease-gene-drug interaction network revealed the potential of key genes as reference markers in T1DM. @*Conclusion@#These results provide new insight into T1DM pathogenesis and novel biomarkers that could be widely representative reference indicators or potential therapeutic targets for clinical applications.

Analysis of results of concurrent hearing and deafness genetic screening and follow up of 33 911 newborns / 中华医学遗传学杂志

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.

Establishment of a vimentin knockout and HIV-1 gp120 transgenic mouse model / 南方医科大学学报

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.

Research Progress of Laportea bulbifera / 中国实验方剂学杂志

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.

The emerging role of nuclear factor erythroid 2-related factor 2 signaling pathway in diabetic chronic complications / 中华内分泌代谢杂志

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.

Application of mouse anti AEG-1 monoclonal fluorescent antibody in malignant serous cavity effusions / 国际检验医学杂志

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 .

Clinical Observation of Piperazine Ferulate Combined with Glutathione in the Treatment of Diabetic Ne-phropathy / 中国药房

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.

Effect on Muc2 gene knockdown in Ht29 cells by CRISPR/Cas9 on probiotics-mediated inhibition of E.coli K1 adhesion and invasion / 南方医科大学学报

<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>

Inhibitory effect of cabozantinib against Listeria monocytogenes invasion in Caco-2 cells in vitro / 南方医科大学学报

<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>

Cabozantinib inhibits Listeria monocytogenes infection in mice / 南方医科大学学报

<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>

Biological characteristics of CD90+ tumor stem cells in ovarian cancer cells / 中国组织工程研究

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.

Prokaryotic expression and purification of anti-AEG-1 single-chain variable antibody / 国际检验医学杂志

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.

Role of CD44 in monocyte transmigration across Cryptococcus neoformans-infected blood-brain barrier in vitro / 南方医科大学学报

<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>

Pathogenesis of uropathogenic Escherichia coli: role of outer membrane protein T and the mechanism / 南方医科大学学报

<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>

Serum level of galectin 3 and its clinical significance in patients with chronic kidney disease / 重庆医学

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 .

The possible relationship between thioredoxin-interacting protein and the pathogenesis of type 2 diabetes mellitus / 中华内分泌代谢杂志

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.

Role of MUC2 gene in the regulation of rat intestinal barrier function by probiotics / 南方医科大学学报

<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>

Construction and functional studies of uropathogenic E. coli strains with ompT gene knockout / 南方医科大学学报

<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>