Distribution and expression of T3SS virulence genes in Pseudomonas aeruginosa and its correlation with drug resistance / 临床检验杂志

Objective@#To investigate the distribution and expression of T3SS virulence genes in clinically isolated Pseudomonas aeruginosa (P. aeruginosa) strains and its correlation with drug resistance. @*Methods@#A total of 68 P. aeruginosa isolates were collected from the First Affiliated Hospital of Wenzhou Medical University in 2015. The antimicrobial susceptibility was detected by the agar dilution method. The distribution of virulence genes such as exoU, exoS, exoT and exoY from different isolates was detected by PCR. The expression levels of transcriptional regulator genes (ptrA and exsA) and effector-related genes (exoT and exoS) in some isolates were determined by real-time fluorescence quantitative PCR, and the Pearson correlation analysis was used to analyze the results. @*Results@#The detection rates of exoT and exoY in 68 P. aeruginosa isolates were higher, accounting for 79.4% and 75.0%, respectively. The detection rate of exoT in wound-sourced isolates was significantly higher than that in sputum (97.0% vs 61.8%, P＜0.01). In addition, the genotype of exoU - /exoS + was the most common, accounting for 51.5% (35/68). The resistance rates of sputum-sourced isolates to imipenem and meropenem were significantly higher than that of wound-sourced isolates (47.1% vs 8.8%, 47.1% vs 14.7%, P＜0.01). The resistance rates of isolates carrying exoU gene to carbapenems, aminoglycosides and fluoroquinolones were higher than those of isolates carrying exoS, exoT or exoY genes. Pearson correlation analysis showed that the expression level of ptrA gene was negatively correlated with those of exoT, exoS and exsA genes (P＜0.05). @*Conclusion@#The P. aeruginosa isolates from our hospital carrying T3SS virulence genes exoT and exoY are common, and the virulence genes are related to the drug resistance of P. aeruginosa. In addition, ptrA may be a potential negative regulatory gene for the expression of T3SS virulence genes.

Clinical efficacy of surgery for chronic subdural hematoma assisted by rigid neuroendoscope / 中国内镜杂志

Objective To discuss the clinical efficacy of surgery for chronic subdural hematoma assisted by rigid neuroendoscope and its surgical techniques. Methods Clinical data of 161 patients with chronic subdural hematoma from August 2009 to December 2015 was analyzed retrospectively. 74 of them experienced surgeries assisted by rigid neuroendoscope (endoscope group) and other 87 cases were operated without neuroendoscope (routine group) during the same period. Results Although there were significant difference in operative duration between the two groups, complications, ratio of total removal of hematoma after surgery, postoperative inpatient duration and recurrent rate of hematoma were more advantageous in endoscope group. The operative duration of endoscope group with (112.68 ± 34.86) min was longer than that of routine group with (74.11 ± 28.23) min (t = 7.75, P = 0.000), while the postoperative inpatient duration of endoscope group with (8.23 ± 2.01) d was shorter than that of another group with (10.79 ± 5.02) d (t = -4.12, P = 0.000). There were no surgical associated complications in endoscope group, but 1 patient in routine group experienced intracerebral hematoma of frontal lobe and associated aphemia. Total removal of hematoma was confirmed in endoscope group with 98.65% (73/74), which was higher than that in routine group with 86.21% (75/78) (χ2 = 8.34, P = 0.004). Hematoma recurrence was found in 16 cases of routine group (18.39%), but more superiority in endoscope group with 1.35% (χ2 = 12.29, P = 0.000). Outpatient follow-up was carried out in all patients from 6 to 38 months with an average duration of 30.06 months. In 17 cases with recurrent hematoma during follow-up, 15 of them were cured by a second surgery, and another 2 patients were cured by atorvastatin. Conclusion As a simple, safe and effective technique, the application of rigid neuroendoscope during surgery for chronic subdural hematoma is more advantage than routine surgery. A self-made suction with adjustable soft curved tip is suitable for such procedure.

Notch1 signaling pathway regulates Th17 cell differentiation and function in murine psoriasis model / 中国免疫学杂志

Objective:To determine the effect of Notch1 signaling pathway on the differentiation and function of Th17 cells in murine psoriasis model.Methods: BALB/c mice were randomly divided into psoriasis model group and control group.Murine psoriasis model was established by topical 5% imiquimod application in combination with intraperitoneal injection of α-2b interferon.The CD4+ T lymphocytes were isolated by magnetic activated cell sorter (MACS).Flow cytometric analysis (FCM) was performed to detect the percentage of Th17 cells.Real-time RT-PCR was employed to measure the mRNA levels of RORγt,IL-17A,Notch1 and Hes-1.The CD4+ T lymphocytes were then divided into γ-secretase inhibitor DAPT groups and control group,and the expression differences of Notch1 signaling molecule and its target gene Hes-1 mRNA levels,Th17 cell percentage,RORγt and IL-17A mRNA levels,and IL-17A concentrations in cell-free supernatant were detected.Results: The expression levels of Th17 cell percentage and RORγt,IL-17A,Notch1 and Hes-1 mRNA in CD4+ T lymphocytes of murine psoriasis model were significantly higher than control mouse[(2.97±0.86)% vs.(0.65±0.11)%,t=15.083;(5.75±0.61) vs.(1.57±0.43),t=21.630;(7.83±0.97) vs.(1.63±0.31),t=25.348;(7.10±1.37) vs.(1.47±0.34),t=17.386;(7.30±1.15) vs.(1.67±0.48),t=18.840,respectively,all P<0.01].Compared with control group,Th17 cell percentage,mRNA expression levels of Notch1,Hes-1,RORγt and IL-17A,and IL-17A concentrations in cell-free supernatant from cultured CD4+ T lymphocytes of murine psoriasis model were dramatically decreased in DAPT treated groups in a dose-dependent way (F=74.368,89.719,126.572,94.558,124.323 and 123.231 respectively,all P<0.01).Conclusion: Notch1 signaling pathway can regulate the differentiation and function of Th17 cells in murine psoriasis model,and may have potential value for the target immunotherapy of psoriasis.

Association of polymorphism in the promoter region of PCA3 gene with risk of prosate cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the polymorphism in the promoter region of PCA3 gene and its relationship with risk of prostate cancer (PCa).</p><p><b>METHODS</b>The promoter region of PCA3 gene of the DNA of peripheral blood mononuclear cells was detected by sequence analysis in the 186 PCa and 141 BPH patients and 135 healthy control individuals. If the samples were detected with polymorphism of insection/deletion, clone sequence analysis was used with pBS-T carrier to verify it.</p><p><b>RESULTS</b>There were 5 polymorphisms. TAAA repeat times: 4, 5, 6, 7, 8, and 8 genotypes (TAAA 4/5, TAAA 4/6, TAAA 5/5, TAAA 5/6, TAAA 5/7, TAAA 5/8, TAAA 6/6, and TAAA 6/7) were detected in the promoter region of PCA3 gene. The eight genotypes were divided into three groups: ≤10TAAA, 11TAAA, ≥12TAAA. Unconditional logistic regression analysis models were used to analyze the relationship between different genotypes and cancer risks adjusted by sex and age. The type 11TAAA and ≥12TAAA was associated with higher relative risk for prostate cancer than the group ≤10TAAA [OR=1.74, 95% CI=1.06-2.87 (for type 11TAAA); OR=5.63, 95% CI=1.85-17.19 (for type ≥12TAAA)]. In the 186 PCa patients, there was 62.4% allele of PCA3 gene with AG/CA mutation found in the promoter 18-19 bp region of PCA3 gene and it had a close relation with the development of prostate cancer.</p><p><b>CONCLUSIONS</b>Short tandem repeats are found in the promoter region of the PCA3 gene in PCa patients, and the increase of TAAA repeat sequences highly enhance the relative risk of prostate cancer development. The occurrence of such STR might be related to the mutations in their upstream loci.</p>

Tumor angiogenesis promoted by fusion of glioma stem/progenitor cells with bone marrow mesenchymal stem cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to clarify whether the fusion of bone marrow mesenchymal stem cells (MSCs) with tumor cells can promote tumor angiogensis.</p><p><b>METHODS</b>Human glioma stem/progenitor cells (GSPCs) (SU3 cells) were transfected with red fluorescent protein (RFP) gene. Bone marrow mesenchymal stem cells (MSCs) were harvested from nude mice with whole-body green fluorescent protein (GFP) gene expression. Then the two kinds of cells were co-cultured in vitro. At the same time SU3-RFP was transplanted into the brain of GFP-expressing nude mice to establish xenograft tumors. The co-cultured cells, GFP/RFP double positive (yellow) cells and blood vessels obtained from the xenograft tumors were observed under fluorescent microscope and laser scanning confocal microscope.</p><p><b>RESULTS</b>After five passages in vitro, MSCs maintained the proliferative activity and highly expressed CD105. CD105 was also expressed in the femurs of GFP-expressing nude mice, tumor cells, blood vessels of SU3 xenograft tumors, and clinical malignant gliomas. When MSCs were co-cultured with SU3-RFP, the ratio of yellow cells co-expressing RFP and GFP was significantly increased after extended time and continuous passages. According to the flow cytometry, yellow cells co-expressing RFP and GFP were 83.7% of the cultured cells. In tissue slices of the xenograft tumors, bundles of yellow vessel-like structure and cross-sectioned yellow vascular wall structures including vascular wall stroma cells were observed with RFP and GFP expression, and were identified as de novo formed vessels derived from fusion of MSCs with SU3-RFP cells.</p><p><b>CONCLUSION</b>Cell fusion occurs between tumor cells and host MSCs and it promotes tumor angiogenesis.</p>

Effects of narrow band ultraviolet B irradiation on the expression of Th17, Treg and their related cytokines in patients with psoriasis vulgaris / 中华物理医学与康复杂志

Objective To study the impact of narrow band ultraviolet B (NB-UVB) irradiation on the expression of T helper 17 (Thl7) and CD4,CD25 and regulatory T (Treg) cells and their related cytokines transfor-ming growth factor beta 1 (TGF-β1) and interleukin-6 (IL-6) to further clarify how NB-UVB treatment helps patients with psoriasis vulgaris.Methods Ninety patients with active stage and stationary stage psoriasis vulgaris (45 cases each) were treated with NB-UVB for 8 weeks.Fifty healthy persons were used as normal controls.Peripheral blood levels of Th17 and Treg were measured by flow cytometry.An enzyme-linked immunosorbent assay (ELISA) was used to measure the serum levels of TGF-β1 and IL-6 before and after treatment.Clinical efficacy was evaluated in terms of the area of psoriasis and severity index (PASI) scores.Restlts Before treatment,the patients showed significantly higher levels of Thl7 cells in their peripheral blood than the controls.Their ratios of Th17 to Treg cells and their serum levels of IL-6 were also significantly higher.The percentage of Treg cells and the serum level of TGF-β1 were significantly lower in the patients.After the NB-UVB treatment,the Th17 cells,the ratio of Th17 to Treg cells and IL-6 had all decreased significantly.The percentage of Treg cells and TGF-β1 levels were significantly high-er compared with before phototherapy.The total effectiveness rate was 86.7％,and the average PASI scores had de-creased significantly.The PASI scores were positively correlated with the percentage of Th17,the Th17 to Treg ratio,and the serum level of IL-6,and negatively correlated with the percentage of Treg cells and TGF-β1.Conclusion The imbalance between Th17 and Treg cells and their cytokines may play an important role in the pathogenesis of pso-riasis.NB-UVB is able to significantly down-regulate the levels of Th17,IL-6 and the progression of Treg levels and TGF-β1 expression.It can regulate the balance between Thl7and Treg cells,which may be one of the mechanisms of NB-UVB treatment for psoriasis.The clinical data demonstrate that NB-UVB treatment is a safe and effective therapy for psoriasis vulgaris.

Value of serum procalcitonin in the predicting prognosis of bacterial infection in critically ill patients / 中华急诊医学杂志

Objective To explore the value of procalcitonin (PCT) in the prediction of the prognosis and severity of bacterial infection in critically ill patients.Methods A total of 116 eligible patients with bacterial infection admitted in the intensive care unit were enrolled in this prospective study from February,2012 through November,2012.Within 24 hours after admission,the serum PCT was determined with immune-chromatography and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score of patients was calculated.Based on the 28-day clinical outcome of patients,the patients were divided into fatal group (n =36) and survival group (n =80).The differences in PCT and APACHE Ⅱ score between the two groups were compared with t test or rank-sum test.The correlation between PCT and APACHE Ⅱ score was determined with Spearman's correlation analysis.Both PCT and APACHE Ⅱ score were analyzed separately and jointly with area under receiver operator characteristic curve (ROC curve,AUC) to predict 28-day survival.Comparison of prediction performance for predicting 28-day survival of patients with bacterial infection between PCT and APACHE Ⅱ was made with U test.Results PCT concentration was significantly higher in fatal group than that in survival group [5.36 (2.07,25) vs.0.24 (1.00,2.14)] (Z =5.596,P ＜0.01).APACHE Ⅱ score within 24 hour after admission was significantly higher in fatal group than that in survival group (24.30 ± 6.71) vs.(16.03 ± 7.23),t =6.147,P ＜ 0.01.Positive correlation between PCT and APACHE Ⅱscore was found to be statistical significance (r =0.388,P＜ 0.01).When rates of 28-day survival in patients were predicted by using PCT and APACHE Ⅱ score,the areas of under curve were 0.804 and 0.792,respectively.AUC of PCT was tenuously larger than that of APACHE Ⅱ score (U =0.2073,P =0.802).Using PCT and APACHE Ⅱ score together to predict 28-day survival,AUC (0.817) was increased.The joint prediction performance was higher than that of either alone,increasing the sensitivity to 90.7％ and the specificity to 75.2％.Conclusions Serum PCT can reflect the severity of the illness and prognosis of infectious disease in the intensive care unit.It can serve as a sensitive marker of predicting 28-day survival.Combining PCT and APACHE Ⅱ score together can increase the prognostic value.

Effects of narrow-band ultraviolet B on expression of IL-23/IL-17 axis and efficacy in patients with psoriasis vulgaris / 中华物理医学与康复杂志

Objective To observe clinical efficacy of NB-UVB in treating psoriasis vulgaris and its effects on expression of serum interleukin-17 (IL-17),interleukin-23 (IL-23) and interleukin-22 (IL-22) in patients with psoriasis vulgaris,and to study the underlying mechanisms of NB-UVB.Methods Ninety patients were recruited and treated with NB-UVB therapy for 8 weeks.Before and after treatment,the serum level of IL-17,IL-23,IL-22,interleukin-l0(IL-10) and transforming growth factor(TGF-β) were tested by use of ELISA method,meanwhile Psoriasis Area and Severity Index (PASI) were used to evaluate clinical efficacy.Fifty healthy volunteers were selected as control group.Results Compared to healthy controls,the level of serum IL-17,IL-23 and IL-22 was significantly higher in patients with psoriasis vulgris (P ＜ 0.01),and IL-10,TGF-β shown lower expression in psoriasis patients (P ＜ 0.01).After 8 weeks of treatment with NB-UVB,serum levels of IL-17,IL-23 and IL-22 in psoriasis patients decreased significantly (P ＜ 0.01),while IL-10,TGF-βelevated significantly (P ＜ 0.01) in contrast.The Psoriasis Area and Severity Index (PASI) results indicated significantly clinical improvement after therapy,and the total effective rate was 87.78％.Conclusion NB-UVB could down-regulate serum IL-17,IL-23,IL-22 and up-regulate IL-10,TGF-β,which may help regulate imbalance of T lymphocytes cells of psoriasis patients.The clinical data demonstrate that the treatment of NB-UVB is a safe,effective method for psoriasis vulgaris.

Analysis of suspicious results of serum HBV DNA detected by fluorescence quantitative PCR / 中华检验医学杂志

Objective To analyze the suspicious results of serum HBV DNA by fluorescence quantitative PCR and develop appropriate countermeasures in order to improve the quality of detection of HBV DNA.Methods Blood samples of patients from the First Affiliated Hospital of Wenzhou Medical College from 2008 to 2011 were analyzed for HBV DNA by fluorescence quantitative PCR.1969 cases of suspicious results,judged by the rule of review the results of serum HBV DNA combined with the historical results,PCR amplification curve,HBV serum markers and clinical diagnosis,were analyzed and redetected by using of two different reagents,careHBV PCR Kit and careHBV PCR Kit V2,at the same time.The consistency and inconsistency ratio of the results were evaluated.Both the reasons of inconsistent and the undetected rates of careHBV PCR Kit were analyzed.The two reasons for the inconsistent results included the reagent related factors,e.g,showing no amplification curve caused by the false negative and abnormal low efficiency of amplification curve,and the non reagent related factors such as operating pollution and other sample factors.Results There were 115 154 blood samples were detected for HBV from 2008 to 2011 and 1969 samples (1.71％) with suspicious results were redetected.The consistency and inconsistency results were 1588 (80.65％) and 381 (19.35％),respectively.Every year from 2008 to 2011,the percentage of the inconsistent results caused by the reagent related factors were 18.87％,20.23％,51.33％ and 59.57％ respectively,which showed an increasing trend,and the percentage of inconsistent results caused by the nonreagent related factors were 81.13％,79.77％,48.67％ and 40.43％ respectively,which showed a declining trend year by year.The undetected rates of careHBV PCR Kit were 2.49％,4.08％,10.09％ and 14.47％ respectively,showing an increasing trend.Conclusions The redetection for the specimens with the suspicious results by using of different reagents can avoid the blind detection of HBV DNA and reduce the experimental error.All the clinical samples for quantitative HBV DNA including the mutations of HBV gene can be measured accurately and effectively,which is helpful to hepatitis B patients for antiviral therapy.

The effects of narrow-band ultraviolet B on the expression of the CC subfamily of chemokines and its efficacy in treating patients with atopic dermatitis / 中华物理医学与康复杂志

Objective To observe the clinical efficacy of narrow-band ultraviolet B (NB-UVB) in the treatment of the patients with atopic dermatitis (AD) and the effects of NB-UVB on the expression of the CC subfamily of chemokines in AD patients. Methods Fifty-five AD patients were treated with NB-UVB with a starting dose of 50％ of the minimal erythma dose.ELISA was used to measure the serum levels of thymus and activation-regulated chemokine ( TARC),cutaneous T cell-attracting chemokine ( CTACK),macrophage-derived chemokine (MDC) and eotaxin in all of the patients before and after treatment,and the clinical efficacy was evaluated.The scores on an atopic dermatitis index (SCORAD) and a visual analogue scale were used for the clinical evaluation.Thirty healthy persons were recruited and served as normal controls. Results The serum levels of TARC,CTACK and MDC were significantly higher in patients with AD than in the normal controls.There was no significant difference between the patients and controls with regard to the average serum level of eotaxin.After treatment with NB-UVB,the serum levels of TARC,CTACK and MDC,but not eotaxin,significantly decreased in the patients.The total clinical effectiveness rate was 76.36％,and the accumulated SCORAD points and VAS scores decreased significantly. Conclusions NB-UVB is able to down-regulate significantly the serum levels of TARC,CTACK and MDC.It can affect immune function and regulate any imbalance of Th1/Th2 cells.This might be one of the mechanisms of NB-UVB treatment for AD.The clinical data demonstrate that NB-UVB is a safe and effective treatment for AD.

Expressions of RORγt and Foxp3 in peripheral blood of children with atopic dermatitis / 中华皮肤科杂志

ObjectiveTo detect the mRNA expression levels of RORγt and Foxp3 genes in peripheral blood mononuclear cells(PBMCs) of children with atopic dermatitis(AD),and to explore their possible roles in the development of AD.MethodsSixty-three children with AD and 54 age-matched normal children were eligible for this study.Real-time fluorescence-based quantitative PCR was carried out to measure the mRNA expressions of RORγt and Foxp3 genes in PBMCs of these subjects.ResultsThe relative expression levels of Foxp3 and RORγt mRNA were significantly higher in children with AD compared with the normal controls (2.4060 ± 0.3355 vs.1.1852 ± 0.4189,t =17.50,P＜ 0.01; 6.9130 ± 0.2046 vs.2.7501 ± 0.2518,t =98.63,P ＜ 0.01 ).A statistical increase was observed in the level of RORγt mRNA in children with severe AD compared with children with mild and moderate AD(7.1203 ± 0.1056 vs.6.8046 ± 0.1731 and 6.8655 ± 0.3671,t =6.61,5.23,both P ＜ 0.01).No significant difference was noted in the expression of RORγt mRNA between children with mild and moderate AD,or in the Foxp3 mRNA expression among children with mild,moderate and severe AD.The severity of AD was positively correlated with the level of RORγt mRNA (r =0.62,P ＜ 0.01 ),but uncorrelated with the level of Foxp3 mRNA(r =0.04,P ＞ 0.05).ConclusionsThe high expressions of RORγt and Foxp3 genes may be involved in the pathogenesis of AD,and the severity of AD is positively correlated with the expression intensity of RORγt gene.

Expression of TLR2 in peripheral blood mononuclear cells and its correlation with interleukin-8 and tumor necrosis factor-α in patients with acne vulgaris / 中华皮肤科杂志

Objective To explore the role of Toll like receptor 2 (TLR2) in the pathogenesis of acne vulgaris. Methods Venous blood was collected from 30 normal human controls and 90 patients with acne vulgaris. The patients were equally divided into three groups, i.e., mild, moderate and severe groups, according to disease severity. Flow cytometry was performed to detect the expression of TLR2 in PBMCs, and double antibody sandwich ELISA to measure the levels of serum interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α). Results A significant increase was observed in the expression of TLR2 in PBMCs, serum level of IL-8and TNF-α in the three groups of patients with acne vulgaris compared with the normal human controls (all P ＜0.01). The expression of TLR2 was positively correlated with the expression of IL-8 (r=0.382, 0.517,0.436,respectively, all P＜0.05) and TNF-α(r=0.641, 0.725, 0.593, respectively, all P＜0.05) in the patients with mild, moderate and severe acne vulgaris, respectively, and with the severity of acne vulgaris (r = 0.406,P＜0.01 ). Conclusion The expression level of TLR2 and related cytokines seems closely correlated with the severity of acne vulgaris.

Effect of whole-process nursing intervention on comfort of patients undergoing radial artery puncture / 中国实用护理杂志

Objective To study the effect of whole-process nursing intervention on comfort of patients undergoing radial artery puncture. Methods 100 patients with radial artery puncture from July, 2007 to June, 2008 were divided into the control group and the experimental group with 50 cases in each group accord-ing to time sequence. Routine whole nursing mode was used in the control group, the whole-process nursing intervention mode based upon routine nursing mode under the instruction of evidence-based method was used in the experimental group. Pain, psychological tensity, body numbness, success rate of radial artery puncture, patients' satisfaction degree were compared between the two groups. Results Every indexes of the experi-mental group were better than those of the control group. Conclusions The whole-process nursing interven-tion mode can promote the comfort degree of patients undergoing radial artery puncture.

Genotype and drug resistance of OXA-51-like carbapenemases in Acinetobaeter baumannii / 中华检验医学杂志

Objective To study the genotypes of OXA-51-like carbepenemases in Aeinetobacter beumannii and its association with drug resistance. Methods The susceptibility of 174 Acinetobacter baumannii against ceftazidime, cefotriaxon, amikacin and ciprofloxacin were detected with disc diffusion method. The minimum inhibitory concentration (MIC) values for meropenem and imipenem were determined with an agar dilution method. VIM, IMP, OXA-23, OXA-24, OXA-51 and OXA-58 β-lactamase genes were determined by PCR. DNA sequencing and genotyping were performed against OXA-51 positivestrains. Results All 174 isolates were negative by PCR for genes OXA-24, OXA-58, IMP and VIM. OXA-23 and OXA-51were amplified in 15.5% (27/174) and 72.4% (126/174) isolates, respectively. Therewere 15.5% (27/174) isolates producing OXA-51-like and OXA-23 carbapenemase simultaneously. Among126 OXA-51-like carbapenemase producing strains, 82.5% (104/126)were OXA-66 genotype, whereas theremaining 17.5% (22/126) strains belong to other genotype. Eight novel OXA-51-like Genotype were foundin this study. Conclusions OXA-66 were the primary genotype of OXA-51-like carbapenemases in A.baumannii. OXA-66 were related to low-level carbapenems resistance and may be associated with resistanceof other drugs. We found new OXA-51-like genotype in clinic isolates of A. baumannii in this study.