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Objective:To investigate changes in skin microecological structures and functions between acute and remission phases in adult patients with severe atopic dermatitis (AD) .Methods:From October 2019 to November 2020, skin scale specimens were collected from 5 body sites (cheeks, cubital fossa, back of the hand, abdomen, lower limbs) of 4 adult patients with severe AD in the acute and remission phases, who visited the outpatient clinic of Guangzhou Institute of Dermatology. The next-generation high-throughput sequencing was performed for metagenomic sequencing to construct the microbial gene catalogue of these specimens, followed by gene annotation and bioinformatics analysis for each sample.Results:A total of 18 phyla, 37 classes, 73 orders, 142 families, 237 genera, and 331 species were identified in the skin specimens from the 4 patients with severe AD. The patients with AD in the remission phase showed significantly increased diversity of skin microbiota and markedly different relative abundance of skin microorganisms compared with those in the acute phase (both P < 0.05). At the microbial species level, Staphylococcus aureus showed the highest impact on the acute phase of AD, while Staphylococcus epidermidis, Moraxella osloensis, Francisella sp., Staphylococcus cohnii, Staphylococcus warneri, Malassezia globosa and Malassezia restricta were enriched in the remission phase of AD with the absolute value of the common logarithm of the linear discriminant analysis score > 2 (Kruksal-Wallis test, all P < 0.05). As KEGG pathway enrichment analysis showed, the differentially abundant genes were annotated into a total of 355 functional pathways, of which 38 pathways were significantly enriched (all P < 0.05), mainly involving Staphylococcus aureus infection, tryptophan metabolism, histidine metabolism, nitrogen metabolism, metabolism of arginine and proline, biosynthesis and degradation of valine, leucine and isoleucine, fatty acid degradation, peroxisome proliferator-activated receptor signaling pathway, etc. Conclusion:The skin microecological structure significantly differed between the acute and remission phases among the patients with severe AD, which may be related to multiple functional pathways, such as Staphylococcus aureus infection, tryptophan metabolism, histidine metabolism and nitrogen metabolism.
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Objective:To compare the detection rate of genital Chlamydia trachomatis (CT) DNA between urine and urethral/cervical swab samples. Methods:From December 2018 to December 2019, a total of 1 475 outpatients were collected from sexually transmitted disease clinics in 7 medical institutions, such as Department of Venereology, Guangzhou Institute of Dermatology, including 1 118 males and 357 females. One urethral/cervical swab sample and one urine sample were collected successively from each patient. Real-time fluorescence-based PCR was performed to detect CT DNA in urine and urethral/cervical swab samples, and paired chi-square test was used to compare the positive rate of CT DNA between the 2 kinds of samples. Random- or fixed-effect meta-analysis was conducted for the test of heterogeneity and merging of positive rates of CT DNA in the urine and urethral/cervical swabs among 7 medical institutions.Results:The positive rate of CT DNA in the urine samples was significantly higher than that in the swab samples from 4 medical institutions (all P < 0.05) , while there was no significant difference in the positive rate of CT DNA between the 2 kinds of samples from 3 medical institutions (all P > 0.05) . The heterogeneity ( I2) estimates of the CT-DNA positive rate in urine and swab samples among different medical institutions were 78.6% (95% CI: 55.9% - 89.6%) and 73.7% (95% CI: 43.7% - 87.7%) , respectively; meta-analysis showed that the total merged positive rate of CT DNA in the urine samples was 10.8% (95% CI: 7.2% - 15.9%) , which was significantly higher than that in the swab samples (7.8%, 95% CI: 4.9% - 12.1%; χ2 = 39.2, P < 0.05) . Compared with the swab sample-based CT-DNA detection method, the sensitivity, specificity, positive predictive value, negative predictive value and consistency rate of the urine sample-based CT-DNA detection method were 97.0% (128/132) , 96.3% (1 293/1 343) , 71.9% (128/178) , 99.7% (1 293/1 297) , and 96.3% (1 421/1 475) , respectively. The positive rate of CT DNA in the urine samples from 1 118 male patients was 11.0% (95% CI: 7.2% - 16.5%) , which was significantly higher than that in the swab samples (7.6%, 95% CI: 4.9% - 11.8%; χ2 = 34.3, P < 0.05) . There was no significant difference in the positive rate of CT DNA between the urine (11.9%, 95% CI: 7.7% - 17.9%) and cervical swab samples from 357 female patients (10.4%, 95% CI: 7.6% - 14.0%; χ2 = 3.2, P > 0.05) . Conclusions:The positive rate of CT DNA in urine samples is higher than or similar to that in urethral/cervical swab samples. The urine sample-based CT-DNA detection method has characteristics of convenience, non-invasiveness, painlessness and low cost, and is worthy of clinical promotion.
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Objective:To investigate the expression of microRNA-155 (miR-155) in serum and kidney of C57BLKS/db (db/db) mice and its role in the pathogenesis of diabetic kidney disease (DKD).Methods:The db/db mice ( n=24) were divided into 6, 8, and 10 weeks old groups ( n=8) with age increasing according to the random number table, and C57BL/6 mice of the same age were used as control group. The expression of miR-155 in mouse serum and kidney tissue was determined using real-time quantitative PCR. The mRNA and protein expression of Ets-1, eNOS, AGTR1 in renal tissues was verified by real-time quantitative PCR, Western blotting and immunohistochemistry. Results:Compared with the control group, the expression of miR-155 in serum of db/db mice at 6, 8 and 10 weeks of age were significantly increased (all P<0.01), and the increase of miR-155 was most obvious at 10 weeks of age ( P<0.01). Meanwhile the expression of miR-155 in kidney tissues of 6, 8 and 10 weeks old db/db mice was significantly up-regulated (all P<0.01), and the highest expression of miR-155 was at 10 weeks of age ( P<0.01). Immunohistochemistry showed that Ets-1, eNOS and AGTR1 were localized in glomerular endothelial cells. The results of real-time quantitative PCR showed that the mRNA expression of Ets-1, eNOS and AGTR1 were down-regulated in the kidney tissues of db/db mice at 6, 8 and 10 weeks of age compared to the control(all P<0.05), and the level of down-regulation was the most obvious at 10 week. Western blotting results showed that there was no significant change in Ets-1, eNOS and AGTR1 in 6-week-old db/db mice compared to the control group; the eNOS protein expression was down-regulated at 8 weeks of age ( P<0.05); the expression of AGTR1 protein was down-regulated ( P<0.05), and the protein expression of Ets-1 and eNOS were significantly down-regulated at 10-week age (both P<0.01). Conclusions:The expression of miR-155 in serum and kidney tissues of db/db mice increases during the progression of DKD, while the expression of miR-155 target genes Ets-1, eNOS and AGTR1 decreases with the progression of DKD. MiR-155 may participate in the development and progression of DKD by inhibiting its target genes Ets-1, eNOS and AGTR1, affecting endothelial cell function.
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Metagenomic analyses of humans and animals have showed that atopic dermatitis (AD) is associated with microbiome dysbiosis in the gut and skin. Decrease of microbial diversity can cause damage to skin barrier and aggravation of AD, and gut microbiome may be involved in the occurrence and development of AD through immune, metabolic and neuroendocrine pathways. This review summarizes the latest advances in the application of metagenomics in tmicrobiological research in and treatment of AD, possible mechanisms underlying microbiome-mediated pathogenesis of AD, and provides a theoretical reference for the microecological therapy of AD.
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Objective To investigate the incidence of renal insufficiency in solitary kidney patients and analyze the risk factors. Methods Patients with solitary kidney who were admitted to the Second Hospital of Lanzhou University from January 2012 to January 2019 were retrospectively selected as subjects. According to estimated glomerular filtration rate (eGFR) level, the patients were divided into two groups: eGFR<60 ml·min-1·(1.73 m2)-1 group and eGFR≥60 ml·min-1·(1.73 m2)-1 group. The data of the general information, laboratory examinations and kidney size were collected, and the differences of the above indicators between the two groups were compared. Logistic regression model was used to analyze the related factors of renal function decline. Results (1) A total of 323 solitary kidney patients with age of (53.8 ± 15.8) years and median duration of 10.0 years were enrolled in the study, including 203 males (62.8％). There were 150 cases (46.4％) with hypertension, 136 cases(42.1％) with proteinuria, and 134 cases (41.5％) with renal insufficiency, even 29 cases(9.0％) had developed into end-stage renal disease. (2) Compared with those in eGFR≥60 ml·min-1·(1.73 m2)-1group, patients in eGFR<60 ml·min-1·(1.73 m2)-1 group had higher age, mean arterial pressure, serum creatinine, serum uric acid, fasting blood glucose, and higher proportion of hypertension and proteinuria, but had lower proportion of congenital solitary kidney, hemoglobin, plasma albumin and residual kidney diameter. The differences of above indicators were statistically significant ( all P<0.05). (3) Logistic regression analysis showed that increasing age (every ten years, OR=1.752, 95％CI 1.455-2.109, P<0.001), anemia (OR=2.327, 95％CI 1.356-3.994, P=0.002), hyperuricemia (OR=5.097, 95％CI 2.873-9.042, P<0.001) and high urine protein level (every 1+, OR=1.515, 95％CI 1.197-1.919, P=0.001) were independent risk factors for renal dysfunction in solitary kidney patients. Conclusions The incidence of renal insufficiency in solitary kidney patients is 41.5％. Patients with solitary kidney may perform varying degrees of kidney damage, such as hypertension, proteinuria and eGFR decline. Increasing age, anemia, hyperuricemia and high urine protein level are independent risk factors for renal insufficiency in solitary kidney patients.
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Objective@#To investigate the incidence of renal insufficiency in solitary kidney patients and analyze the risk factors.@*Methods@#Patients with solitary kidney who were admitted to the Second Hospital of Lanzhou University from January 2012 to January 2019 were retrospectively selected as subjects. According to estimated glomerular filtration rate (eGFR) level, the patients were divided into two groups: eGFR<60 ml·min-1·(1.73 m2)-1 group and eGFR≥60 ml·min-1·(1.73 m2)-1 group. The data of the general information, laboratory examinations and kidney size were collected, and the differences of the above indicators between the two groups were compared. Logistic regression model was used to analyze the related factors of renal function decline.@*Results@#(1) A total of 323 solitary kidney patients with age of (53.8±15.8) years and median duration of 10.0 years were enrolled in the study, including 203 males (62.8%). There were 150 cases (46.4%) with hypertension, 136 cases(42.1%) with proteinuria, and 134 cases (41.5%) with renal insufficiency, even 29 cases(9.0%) had developed into end-stage renal disease. (2) Compared with those in eGFR≥60 ml·min-1·(1.73 m2)-1group, patients in eGFR<60 ml·min-1·(1.73 m2)-1 group had higher age, mean arterial pressure, serum creatinine, serum uric acid, fasting blood glucose, and higher proportion of hypertension and proteinuria, but had lower proportion of congenital solitary kidney, hemoglobin, plasma albumin and residual kidney diameter. The differences of above indicators were statistically significant (all P<0.05). (3) Logistic regression analysis showed that increasing age (every ten years, OR=1.752, 95% CI 1.455-2.109, P<0.001), anemia (OR=2.327, 95% CI 1.356-3.994, P=0.002), hyperuricemia (OR=5.097, 95% CI 2.873-9.042, P<0.001) and high urine protein level (every 1+, OR=1.515, 95% CI 1.197-1.919, P=0.001) were independent risk factors for renal dysfunction in solitary kidney patients.@*Conclusions@#The incidence of renal insufficiency in solitary kidney patients is 41.5%. Patients with solitary kidney may perform varying degrees of kidney damage, such as hypertension, proteinuria and eGFR decline. Increasing age, anemia, hyperuricemia and high urine protein level are independent risk factors for renal insufficiency in solitary kidney patients.
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Objective@#To identify differentially expressed genes in the transcriptome of the lesional versus nonlesional skin tissues of patients with moderate and severe atopic dermatitis (AD) , and to elucidate their roles in the pathogenesis of AD.@*Methods@#From July to October in 2016, lesional and nonlesional skin tissues were obtained from 5 outpatients of Han nationality with AD in Guangzhou Institute of Dermatology, Institute of Dermatology, Guangzhou Medical University. The next-generation high-throughput transcriptome-wide RNA sequencing (RNA-seq) was performed to identify differentially expressed genes, which were subjected to GO function annotation and KEGG pathway analysis. Real-time fluorescence-based quantitative PCR (qRT-PCR) was conducted to verify differences in candidate gene expression between lesional and nonlesional skin tissues.@*Results@#An average of 10.96 GBs sequence reads were acquired among 10 samples. A total of 21 729 genes were detected, including 19 268 known genes and 2 545 predicted novel genes. A total of 23 153 new transcripts were detected, of which 18 889 were new alternative splicing subtypes of known protein-coding genes, 2 545 were transcripts belonging to new protein-coding genes, and the remaining 1 719 belonged to long-stranded non-coding RNA. Totally, 78 differentially expressed genes were identified between the lesional and nonlesional skin tissues, including 69 upregulated and 11 downregulated genes in the lesional skin tissues. Among them, there were several genes known to be associated with AD inflammation (CXCL1/2/8, IL6/IL1β, MMP1, SERPINB4, S100A2, GZMB, OASL, OSM) and barrier (KRT16, FABP5, CYP1A1) and keratinocyte differentiation (IL-20) . GO analysis revealed that functions of 72 differentially expressed genes could be annotated. KEGG pathway analysis showed that the differentially expressed genes were grouped into 132 signaling pathways, of which 13 were significantly enriched, including the interleukin (IL) -17 pathway, NOD-like receptor signaling pathway, Toll-like receptor signaling pathway, etc. qRT-PCR showed that the mRNA expression levels of candidate genes CXCL1, KRT6A, IL36A, SERPINB4 and PSAPL1 was consistent with the transcriptome sequencing results.@*Conclusions@#Differentially expressed genes and related important regulatory signaling pathways were identified between the lesional and nonlesional skin tissues of patients with AD at the transcriptional level, and the IL-17 pathway was found to be mostly enriched in AD lesions in patients of Han nationality. These findings provide an important basis for further study on the pathogenesis of AD..
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Objective To identify differentially expressed genes in the transcriptome of the lesional versus nonlesional skin tissues of patients with moderate and severe atopic dermatitis(AD), and to elucidate their roles in the pathogenesis of AD. Methods From July to October in 2016, lesional and nonlesional skin tissues were obtained from 5 outpatients of Han nationality with AD in Guangzhou Institute of Dermatology, Institute of Dermatology, Guangzhou Medical University. The next-generation high-throughput transcriptome-wide RNA sequencing (RNA-seq) was performed to identify differentially expressed genes, which were subjected to GO function annotation and KEGG pathway analysis. Real-time fluorescence-based quantitative PCR(qRT-PCR)was conducted to verify differences in candidate gene expression between lesional and nonlesional skin tissues. Results An average of 10.96 GBs sequence reads were acquired among 10 samples. A total of 21729 genes were detected, including 19268 known genes and 2545 predicted novel genes. A total of 23153 new transcripts were detected, of which 18889 were new alternative splicing subtypes of known protein-coding genes, 2545 were transcripts belonging to new protein-coding genes, and the remaining 1719 belonged to long-stranded non-coding RNA. Totally, 78 differentially expressed genes were identified between the lesional and nonlesional skin tissues, including 69 upregulated and 11 downregulated genes in the lesional skin tissues. Among them, there were several genes known to be associated with AD inflammation (CXCL1/2/8, IL6/IL1β, MMP1, SERPINB4, S100A2, GZMB, OASL, OSM) and barrier (KRT16, FABP5, CYP1A1) and keratinocyte differentiation (IL-20). GO analysis revealed that functions of 72 differentially expressed genes could be annotated. KEGG pathway analysis showed that the differentially expressed genes were grouped into 132 signaling pathways, of which 13 were significantly enriched, including the interleukin(IL)-17 pathway, NOD-like receptor signaling pathway, Toll-like receptor signaling pathway, etc. qRT-PCR showed that the mRNA expression levels of candidate genes CXCL1, KRT6A, IL36A, SERPINB4 and PSAPL1 was consistent with the transcriptome sequencing results. Conclusions Differentially expressed genes and related important regulatory signaling pathways were identified between the lesional and nonlesional skin tissues of patients with AD at the transcriptional level, and the IL-17 pathway was found to be mostly enriched in AD lesions in patients of Han nationality. These findings provide an important basis for further study on the pathogenesis of AD. .
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OBJECTIVE@#To investigate the expression profile of miR-122-5p in melanoma tissues and the effect of miR-122-5p on the proliferation, cell cycle and apoptosis of human melanoma cell lines SK-MEL-110 and A375.@*METHODS@#The expression profiles of miR-122-5p in melanoma and pigmented nevus tissues were detected using real-time fluorescence quantitative PCR (qRT-PCR). SK-MEL-110 and A375 cells transfected with miR-122-5p inhibitor or negative control inhibitor (NC) I were examined for miR-122- 5p expression using qRT-PCR and changes in cell proliferation, cell cycle and apoptosis using MTT assay or flow cytometry. NOP14 mRNA and protein expressions in the cells were detected using qRT- PCR and Western blotting, respectively. Luciferase reporter assay was used to confirm the identity of NOP14 as the direct target of miR-122-5p.@*RESULTS@#The relative expression of miR-122-5p in human pigmented nevus tissues and melanoma tissues was 1.23±0.270 and 7.65 ± 1.37, respectively. The relative expression of miR-122-5p in SK-MEL-110 and A375 cells transfected with miR-122-5p inhibitor was 0.21 ± 0.08 and 0.17 ± 0.05, respectively. miR-122-5p inhibitor obviously inhibited the cell proliferation and increased the percentage of cells in G1 stage in both SK-MEL-110 and A-375 cells, but did not cause obvious changes in the apoptosis of the two cells. miR-122-5p inhibitor did not significantly affect the expression level of NOP14 mRNA, but obviously increased the expression level of NOP14 protein. Luciferase reporter assay revealed a significantly lower luciferase activity in cells co-transfected with miR-122-5p mimics and wild-type psi-CHECK2-3'UTR plasmid than in the cells cotransfected with NC and wild-type psi-CHECK2-3'UTR plasmid (0.21 ± 0.14 0.56 ± 0.1, < 0.01).@*CONCLUSIONS@#miR-122-5p expression is upregulated in melanoma tissues, indicating its involvement in the development of melanoma. miR-122-5p inhibits the proliferation of SK-MEL-110 and A-375 cells possibly by affecting the cycle through NOP14.
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Humans , Apoptosis , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Luciferases , Metabolism , Melanoma , Metabolism , Pathology , MicroRNAs , Metabolism , Neoplasm Proteins , Metabolism , Nevus, Pigmented , Metabolism , Pathology , Nuclear Proteins , Metabolism , Skin Neoplasms , Metabolism , Pathology , Up-RegulationABSTRACT
Objective To investigate the effects of Ligustilide on the withdrawal syndromes syn-dromes and monoamine neurotransmitters of hypothalamus and nucleus accumbens in morphine-dependent rats. Methods Totally 60 SD rats were divided into control group,model group,clonidine group and Ligust-ilide high(80 mg/kg),medium(40 mg/kg) and low(20 mg/kg) dose group according to the random number table with 10 in each group. Rats were given in gradual increasing doses of morphine to produce physical de-pendence. Morphine withdrawal syndrome was precipitated by naloxone and withdrawal symptoms were evalu-ated by Ryuta Tomoji score. The level of norepinephrine ( NE), dopamine ( DA) and 5-hydroxytryptamine (5-HT) in rats were tested with enzyme-linked immunosorbent assay(ELISA). Results The total score of somatic withdrawal syndromes in the control group,model group,clonidine group and Ligustilide low,medium and high dose group were 0,(31. 83±7. 33),(17. 92±6. 88),(25. 58±5. 99),(19. 88±4. 82) and (16. 75 ±4. 01) . Compared with the model group,the morphine withdrawal syndromes scores of Ligustilide low,me- dium and high dose groups and clonidine group were reduced(all P<0. 05). The level of NE,DA and 5-HT in hypothalamus and nucleus accumbens were increased compared with that of control group. Compared with the model group,the level of NE,DA and 5-HT in hypothalamus and nucleus accumbens of Ligustilide low, medium and high dose groups and clonidine group were significantly reduced (P<0. 05). Conclusion Ligu-stilide can effectively alleviate the symptoms in morphine-withdrawal rats,which may be related to the inhibi-tion of excessive release of monoamine neurotransmitters in hypothalamus and nucleus accumbens.
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Objective To investigate the protective effect and mechanism of garlicin on oxidative stress injury of human melanocytes.Methods There were blank group,control group,hydrogen peroxide group,garlicin group,experiment 1,2 and 3 groups.No cells in the blank group were only added with complete culture medium.The control group was added with complete medium;0.4 mmol/L hydrogen peroxide complete medium was added to the hydrogen peroxide group.Garlicin group was added with freshly prepared garlicin complete culture medium with concentration of 40 μmol/L;Experiment groups 1,2,and 3 were treated with different concentrations of garlicin (80,40,and 20 μmol/L garlicin complete medium,respectively) to interfere with melanocytes treated with 0.4 mmol/ L hydrogen peroxide during logarithmic growth period.After 24 h of drug intervention,the cell morphology was observed under an inverted microscope.The protective effect of garlicin on melanocytes damaged by oxidative stress was measured by MTT colorimetric method.Results Different concentrations of garlicin had different protective effects on melanocytes induced by hydrogen peroxide.It could be concluded that the activity of melanocytes in hydrogen peroxide group decreased significantly (43.610 ± 3.872)％ (P＜0.05),but there was no statistical difference between the two groups (P=0.345).The activity of melanocytes in experimental group 1 was significantly decreased (58.223 ± 2.806) ％ but higher than that in experimental groups 2 and 3 (P＜0.05).Conclusions Allicin inhibits the production of intracellular ROS in human melanocytes induced by H2 O2,regulates oxidative stress in human melanocytes,and counteracts H2O2-induced apoptosis.Therefore,allicin may be a protective factor in mediating oxidative stress in the body.
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Stroke remains the major complication among patients on dialysis.In chronic hemodialysis patients,prevalence and incidence of stroke are higher than those in general population.This article provides an overview of stroke in patients on dialysis,including clinical features,early warning and recognition,prevention and treatment.
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Objective To determine the expression of Caspase 8 and phospho-Akt(p-Akt)in condyloma acuminatum(CA)lesions, and to evaluate their significance. Methods Skin lesion samples were collected from 30 patients with CA, cancer tissue samples from 20 with cervical cancer, and normal skin samples from 20 healthy controls. All the samples were subjected to paraffin embedding. An immunohistochemical study was conducted to determine the expression and distribution of Caspase 8 and p-Akt in the above samples. Results The expression rate of Caspase 8 was significantly lower in CA lesions (23.33%)than in normal skin samples(90%, P < 0.01)and cervical cancer lesions(80%, P < 0.001). Moreover, the expression rate of p-Akt in CA lesions(93.33%)was significantly higher than that in the normal skin samples(90%, P<0.001), but lower than that in the cervical cancer lesions(95%, P<0.001). No significant correlations were observed between the expression of Caspase 8 and p-Akt in either CA lesions or normal skin samples. However, the expression of Caspase 8 was positively correlated with the expression of p-Akt in cervical cancer lesions(r=0.369, P<0.05). Conclusion Both suppressed apoptosis initiation of Caspase 8 and anti-apoptotic effect of p-Akt may be involved in the occurrence and development of CA.
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Objective To investigate the relationship between carotid artery intima-media thickness and renal function in patients with diabetes mellitus.Methods 424 patients of type 2 diabetes without dialysis were enrolled in a cross-sectional study.According to their artery intima-media thickness (IMT),the patients were divided into normal group and higher IMT group.All patients according to UAER or 24h urinary protein were divided into normal proteinuria group,micro-proteinuria group and clinical proteinuria group.The biochemical examination,eGFR,and atherosclerotic plaque of different groups were compared.Pearson or spearman correlation was used to analyze the relationship between eGFR,IMT and other parameters.Risk factors for eGFR decline were analyzed by binary logistic regression.Results Compared with normal group,patients in the higher IMT group were older [(63.3±10.2) year vs (52.5 ± 10.6) year,P ＜ 0.05],and underwent longer duration of diabetes [(8.9±6.7) year vs (6.2±5.7) year,P ＜ 0.05].Their level of eGFR was decreased [(75.92±28.00) ml/min vs (91.64±24.05) ml/ min,P ＜ 0.05],while plaque incidence (71.3％ vs 18.3％,x2=112.42,P ＜ 0.01) and prevalence of hypertension (56.4％ vs 29.6％,x2=27.22,P ＜ 0.01) increased.Correlation analysis showed that IMT was positively correlated with age (r=0.503,P ＜ 0.01),duration of diabetes (r=0.204,P ＜ 0.01),24 h urine protein (rs=0.175,P ＜ 0.05),plaque (rs=0.562,P ＜ 0.01),and hypertension (rs=0.193,P ＜ 0.01),but negatively correlated with eGFR (r=-0.307,P ＜ 0.01).Logistic regression analysis showed that age,serum uric acid,24 h urine protein and carotid artery intima-media thickness were independent risk factors for eGFR decline [OR=1.115,95％CI(1.053,1.165),P ＜ 0.001;OR=1.008,95％CI (1.002,1.014),P=0.006;OR=1.492,95％ CI(1.170,1.903),P=0.001;OR=1.619,95％ CI(1.121,2.339),P=0.010].Conclusion Carotid artery intima-media thickness is an independent risk factor for kidney function decline in patients of diabetes.
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Objective To investigate the regulatory effects of miR-145 on the proliferation,cell cycle and apoptosis of a human keratinocyte cell line HaCaT.Methods miR-145 mimics and negative control (NC) mimics were chemically synthesized and then transiently transfected into HaCaT cells respectively.After additional culture for different durations,real-time PCR was performed to determine the expression level of miR-145,MTS assay to estimate cell proliferation,and flow cytometry to detect cell apoptosis and cycle.Luciferase assay,real-time PCR and Western blot were conducted to determine whether NRAS was the target gene of miR-145.Results The miR-145 expression level in miR-145 mimic-transfected cells increased by 85.00 ± 1.21 folds compared with NC mimic-transfected cells (t =115.90,P ＜ 0.0001).The transfection with miR-145 mimics significantly inhibited the proliferation of HaCaT cells (F =8.76,P =0.008),and the inhibitory effect significantly varied with the duration (24-96 hours) of culture after transfection,with no interaction effect between the transfection with miR-145 mimics and culture duation (F =1.21,P =0.18).Compared with NC mimic-transfected cells,those transfected with miR-145 mimics showed a significant increase in the proportion of early apoptotic cells (18.9％ ± 4.1％ vs.4.3％ ± 1.2％,t =7.126,P ＜ 0.01),late apoptotic cells (9.3％ ± 2.3％ vs.3.6％ ± 1.6％,t =12.38,P ＜ 0.01),G1-phase cells (85.83％ ± 5.2％ vs.62.08％ ± 6.23％,t =11.78,P =0.007),but a significant decrease in the percentage of G2-phase cells (6.26％ ± 1.2％ vs.19.36％ ± 3.45％,t =7.610,P =0.017) and S-phase cells (7.91％ ± 1.3％ vs.18.56％ ± 5.23％,t =7.230,P=0.019).As luciferase assay showed,luciferase activity was significantly lower in HaCaT cells cotransfected with miR-145 mimics and a recombinant luciferase reporter vector psi-CHECK2-NRAS-wild carrying the wild-type 3'UTR of NRAS than in those cotransfected with NC mimics and the vector psi-CHECK2-NRAS-wild (t =11.09,P =0.008),but similar between cells cotransfected with miR-145 mimics and a recombinant luciferase reporter vector psi-CHECK2-NRAS-mut carrying the mutant-type 3'UTR of NRAS and those cotransfected with NC mimics and the vector psi-CHECK2-NRAS-mut (P ＞ 0.05).Real-time PCR and Western blot revealed that the overexpression of miR-145 mimics had no significant effect on NRAS mRNA expression (P ＞ 0.05),but significantly inhibited NRAS protein expression (1.52 ± 0.07 vs.0.20 ± 0.02,t =28.43,P＜ 0.01).Conclusion miR-145 might inhibit proliferation and promote apoptosis of HaCaT cells by influencing cell cycle via NRAS.
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Objective To investigate the expression and clinical significance of serum microRNA (miRNA) expression profiling in the occurrence and progression of diabetic nephropathy.Methods The miRNA expression profiling was detected by miRNA TaqMan Low Density Array chip from 10 patient with diabetic nephropathy,10 diabetes patients with normoalbuminuria and 10 health control.Real-time quantitative PCR was applied to verify the result of miRNA array in serum samples of 66 patients with diabetic nephropathy (36 patients with microalbuminuria,30 patients with macroalbuminuria),40 diabetes patients with normoalbuminuria and 40 health control.And the relationship of differetial expression with clinical features was analyzed.Results miR-150-5p,miR-155-5p,miR-30e-5p and miR-3196 being validated by real-time quantitative PCR differentially expressed in 3 groups of serum samples from the diabetes patients with microalbuminuria (n=36),with normoalbuminuria (n=40) and health control (n=40) (P ＜ 0.05).Serum miR-150-5p (P=0.005) and miR-155-5p (P=0.006) changed significantly between diabetes patients with microalbuminuria (n=36) and with macroalbuminuria (n=30).Compared with the diabetes patients with microalbuminuria,serum miR-150-5p and miR-155-5p increased by 2.3 and 1.5 times in macroalbuminuria group,respectively.Estimated glomerular filtration rate and urinary albumin excretion rate significantly correlated with serum miR-150-5p and miR-155-5p level.Conclusions miR-150-5p and miR-155-5p may be involved in the process of pathological mechanisms of diabetic nephropathy.Serum miR-150-5p and miR-155-5p may be regarded as potential biomarkers to diagnosis the occurrence and development of diabetic nephropathy.
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BACKGROUND:Hemodialysis therapy is an important means for the treatment of acute renal failure, which aims to remove excess water and toxins and maintain acid-base balance of a patient, creating conditions for medication and nutrition therapy while avoiding multiple organ failure. OBJECTIVE:To compare bicarbonate-and lactate-buffered solutions for acute continuous hemodiafiltration in acute renal failure. METHODS:A computer-based search was performed in PubMed, EMBASE, SCI, Cochrane Library, Chinese Biomedical Literature Database, China Journal Ful Text Database, Chinese Medical Association Journals for randomized control trials related to bicarbonate-versus lactate-buffered solutions for hemodiafiltration in acute renal failure published before January 2014. The quality of the included studies was evaluated by Cochrane Handbook, and data were analyzed by RevMan 5.1 from the Cochrane Col aboration. RESULTS AND CONCLUSION:Four studies (171 patients) met inclusion criteria. Overal , patients treated with bicarbonate-buffered solutions had fewer cardiovascular complications and symptomatic hypotension events as wel as lower serum lactate levels than patients who received lactate-buffered solutions (P<0.05). There were no differences in mortality, serum bicarbonate levels, serum creatinine, serum pH, carbon dioxide partial pressure. The current evidence shows that patients undergoing bicarbonate-buffered solutions may experience fewer cardiovascular complications and symptomatic hypotension. Given the limited research, it is insufficient to recommend for clinical use.
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Objective To investigate the characteristics and risk factors of anemia in renal transplant recipients over 60 years old.Methods Clinical data of one hundred and sixty-eight renal transplant recipients over 60 years old were retrospectively analyzed.Logistic regression analysis was used to determine the risk factors of anemia.Results In 168 cases of renal transplant recipients,the incidence of anemia was 45.2％(76/168).Forty cases were normocyte and normochromic,26 cases were microcytic hypochromic,10 cases were hemolytic anemia.In these anemic recipients,51 cases were short of erythropoietin (EPO),25 were EPO resistance.The incidence of malnutrition and cardia-cerebrovascular complication was higher in recipients with anemia than those without anemia (P＜0.01).The incidence of anemia in CsA+Aza+Pred treatment was 57.1％,which was significantly higher as compared to other three treatments (P＜0.01).Unconditional multivariate Logistic regression analysis revealed that male,creatinine level,acute reject reaction,delayed graft function (DGF) were the independent predictors of anemia,the corresponding OR values were 1.089,5.156,6.345,1.876.Conclusions Anemia is a common and serious complication in renal transplant recipients over 60 years old.Male,creatinine level,acute reject reaction,DGF are the independent risk factors of anemia in renal transplant recipients over 60 years old.
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Objective To investigate the incidence and risk factors of acute kidney injury (AKI) after craniocerebral injury.Methods A single cohort of 791 patients who suffered from craniocerebral injury from January 2008 to January 2010 in the Second Hospital of Lanzhou University were prospectively analyzed.Craniocerebral injury was defined according to definite medical history of craniocerebral injury,the verification of CT and Glasgow coma scale (GCS) score.AKI was defined as a relative 50％ increase or an absolute increment of 26.4 μmol/L in Scr within 48 hours and/or urine volume ＜0.5 ml·kg-1·h-1 up to 6 h.Multivariate Logistic regression analysis was used to evaluate possible risk factors associated with post-craniocerebral injury AKI.Results Of the 791 patients,the incidence of AKI was 39.4％.In hospital mortality of AKI patients was 27.9％,which was 5.065 times of non-AKI patients (P＜0.01).The incidence of AKI in patients with lower GCS score (≤8 score,heavy group)was 69.7％,which was significantly higher as compared to moderate and mild groups (P＜0.01).Unconditional multivariate Logistic regression analysis revealed that lower GCS score (≤ 8 score),hypotension (systolic pressure＜90 mm Hg),elderly and male were the independent predictors of AKI episodes,the corresponding OR values were 2.932,2.176,1.789,1.544 respectively.Conclusions AKI is a common complication after craniocerebral injury.Lower GCS score,hypotension,elderly and male are the independent risk factors of AKI in patients after craniocerebral injury.
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Objective To assess the human β-defensin-2 ( hBD2 ) gene therapeutic efficacy in a rat urinary tract infection (UTI) model via intravesical liposome-mediated gene transfer.Methods Fifty-six female Wistar rats (class SPF) weighting 1 80 -220 were randomly divided into an experiment group and a control group ( each n =28 ).The animals were administered either 250 μl recombinant pCAGG-hBD2 or control vector pCAGG intravesically.After 48 h,rats in both groups were infected via intravesical inoculation with 200 μl of the bacterial suspension of UTI89 ( 1 × 108 CFU/ml).The rats were sacrificed at 4,24,48,and 72 h post-inoculation.The bladders were aseptically removed and bisected.One half was fixed in neutral buffered formalin for histological analysis.The remaining half-bladders were homogenized and titered for surviving bacteria.The clean-catch urine sample from each rat was collected in sterile before they were killed for bacterial titers determined and WBC counted.Results Numbers of bacterial colony-forming unit in urine and bladders from hBD2 gene treated UTI rats were significantly lower than those from the control vector administered UTI rats at 24,36,and 72 h after infection ( P ＜ 0.05 ).The amount of WBC in urine was significantly less in the defensin group than in the control group.In addition,in vivo expression of hBD2 could reduce mucosa damage,interstitium edema and inflammatory cells infiltration in UTI animals.Conclusions The successful inhibition of UTI progression could be obtained with hBD2 gene therapy.Recombinant beta-defensin-2 could kill UTI89 in vivo and suppress the subsequent infection-induced inflammatory responses.