60,≤60 year age groups and control group was 2.49?1.82,1.82?1.58,0.90?1.38, significantly decreased(P

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.

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.

BACKGROUND:The establishment of a safe, reliable and easily repeatable mouse model of nonalcoholic fatty liver disease is the prerequisite for the study of the diagnosis and treatment of the disease.
OBJECTIVE:To establish a C57BL/6 mouse model of nonalcoholic fatty liver disease and observe changes of biochemical indicators, which can provide a theoretical basis for its pathogenesis and drug treatment.
METHODS:Sixty healthy male C57BL/6 mice were randomly divided into a control group of 30 cases (normal diet), and a model group of 30 cases (high fat diet). Models of nonalcoholic fatty liver were established. At 8 weeks, body mass, liver index, and homogenate superoxide dismutase activity in the liver were detected. Changes in serum alanine aminotransferase, aspartate aminotransferase, triglyceride glycerol, cholesterol, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol were observed. Pathological examination was performed.
RESULTS AND CONCLUSION:(1) Pathological sections showed that large droplets and smal lipid droplets in the mouse liver and spread the whole liver. Swel ing of the liver cel s, visible cytoplasmic vacuoles and obviously inflammatory changes in liver cel s were observed in the model group. (2) Body weight and liver index were significantly higher in the model group than in the control group (P<0.05). Superoxide dismutase activity was significantly reduced in the liver (P<0.05). (3) Triglycerides, cholesterol, and low-density lipoprotein cholesterol levels were significantly higher, but high-density lipoprotein cholesterol levels were significantly lower in the model group than in the control group (P<0.05). (4) Nonalcoholic fatty liver mouse model is ideal for high-fat diet-induced animal model. The method is simple, repetitive, and can provide a stable animal model for the study on the mechanism of nonalcoholic fatty liver disease and drug treatment.

An ultra-high performance liquid chromatography-mass spectrometry ( UPLC-MS/MS) method was developed for the determination of 8 kinds of cephalosporins, cefoperazone, cefquinome, cefalonium, cefazolin, cefapirin, Ceftiofur, cefpirome and cefalexin, in edible parts of aquatic products. The samples were extracted with acetonitrile-water and cleaned up by multi-walled carbon nanotubes ( MwCNTs) SPE cartridge. All the target compounds were separated on an Acquity Xselect CSH C18 column with gradient elution by using acetonitrile and 0. 1% formic acid aqueous as eluent, and detected by UPLC-MS/MS under ESI+ ionization and MRM mode. Under optimized conditions, this method had a good linearity (R2≥0. 995) and the limits of quantification were in the range of 2-10 μg/kg ( S/N=10 ) . The recoveries of the method for the target compounds spiked at three different levels were 67. 3%-94. 2% with the relative standard deviations (RSDs) of 3. 3%-14%. The method had the characteristics of low cost, high accuracy and good precision, and could meet the requirements of cephalosporins determination.

A method was developed for the determination of tetrodotoxin in marine organisms by high perfor-mance liquid chromatography-mass spectrometry with immunoaffinity column. The samples were extracted with 1% acetic acid methanol solution and diluted with phosphate buffer at pH 7-8. After cleaned up by immuno-affinity column, the samples were analyzed by LC-MS/MS and quantitatively determined by external standard method. The chromatographic separation was performed on an ACQUITY UPLC BEH Amide column with gradient elution by using acetonitrile and 5 mol/L ammonium acetate solution containing 0. 1% formic acid as mobile phase. Detection was carried out by electrospray positive ionization mass spectrometry in the multiple reaction monitoring mode. Linear ranges of TTX was in the range of 0. 3 -20. 0 μg/L with correlation coeffi-cient more than 0. 997. The quantification limit of the method was 0. 3 μg/kg. The recoveries of standard addition for tetrodotoxin were 88. 7%-102. 3%, and the relative standard deviation was 2. 0%-6. 4%. The method could be used to identify and quantify tetrodotoxin in marine organisms with satisfactory reproducibility and sensitivity.

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.

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.

Objective To explore the accurate positioning of Kawase triangle area via intradural anterior subtemporal transpetrosal approach. Methods On 14 dry skulls, the highest point of arcuate eminence (A), the junction of lateral margin of petrous ridge and anterior margin of transverse sinus (J), the petrous apex (P), the outermost point of sulcus nervi petrosi superficialis majoris (B), the outermost point of foramen spinosum (C), the outermost point of foramen ovale (D), the outermost point of trigeminal notch (E), and the outermost point of foramen lacerum (F) were marked. The distances of JA, JB, JC, JD, JE and JF were measured using point J as reference point. Using the line (JP) between point J and point P as the baseline, the angles of baseline with JA, JB, JC, JD, JE, and JF were measured. Results There were no significant differences in the distances of JA, JB, JC, JD, JE and JF between the left and right sides (P>0.05). There were no significant differences in the angles of the baseline with JA, JB, JC, JD, JE, and JF between left and right sides (P>0.05). Conclusion Using point J and the baseline JP as referent indexes, bony structures can be precisely located via intradural anterior subtemporal transpetrosal approach to orientate the Kawase triangle area; this method can insure rapid, accurate and safe drilling of anterior petrosal bone and exposing of petroclival region.

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.