Objective To observe the effects of butylphthalide on the expression of autophagy-related protein and mRNA in l-meth-yl-4-phenyl-pyridiniumion (MPP+)-induced SH-SY5Y cells, and to explore the protective effect and possible mechanism of butylphthalide to the cell model of Parkinson's disease. Methods The SH-SY5Y cells were divided into control group (A), MPP+group (B), rapamycin pre-treated+MPP+group (C) and Butylphthalide pretreated+MPP+group (D). The relative viability of SH-SY5Y cells induced by MPP+was measured with MTT assay, the morphology of SH-SY5Y cells was observed. The expression of microtubule associated protein 1 light chain 3 (LC3)-II/I and Beclin 1 protein was detected by Western blotting. And the expression of LC3-II/I and Beclin 1 mRNA were assayed by re-al-time quantitative reverse transcription-PCR (RT-PCR). Results The viability rates of cells were significantly lower in group B than in group A (t=20.270, P<0.001), and were significantly higher in groups C and D than in group B (t>8.770, P<0.001), however, there was no significantly difference between groups C and D (t=2.270, P=0.064). The expression of LC3-II/I and Beclin 1 was higher in group B than in group A (t>6.647, P<0.01), and was higher in groups C and D than in group B (t>3.630, P<0.01), however, there was no significantly differ-ence between groups C and D (t<2.238, P≥0.05). Conclusion Butylphthalide could prevent the injury of SH-SY5Y cells induced by MPP+, which may affect Parkinson's disease by inducing autophagy.

Objective To investigate the distribution of human thyroid stimulation hormone(hTSH) in dried venous blood spots of neonatal screening for congenital hypothyroidism in Guilin ,and establish the hTSH cut‐off value for neonatal screening .Methods We selected 101 817 cases of neonatals with the local household registration from January 2011 to December 2013 ,and collected their venous blood to be made into dried venous blood spots on filter paper ,then the hTSH levels in dried blood spots were deter‐mined for congenital hypothyroidism ,According to screening results ,using the percentile method to calculate the hTSH P50 ,P99 of all the normal neonatals and neonatals that born in different quarters .Results Among 101 817 neonatals who were screened ,101 746 cases of normal neonatals ,71 cases of patients(including 47 cases of patients with CH ,24 cases of patients with high hTSH hy‐perlipidemia) ,the hTSH levels of normal neonatals showing a skewed distribution ,P50 ,P99 were 2 .45 μU/mL ,9 .86 μU/mL .the hTSH levels of the neonatal who were born in different quarters was changed with the change in the quarter ,P50 ,P99 were 2 .15μU/mL ,9 .20 μU/mL in The Spring ;P50 ,P99 were 1 .97 μU/mL ,8 .38 μU/mL in the summer ;P50 ,P99 were 2 .67 μU/mL ,10 .43μU/mL in the autumn ;P50 ,P99 were 2 .90μU/mL ,11 .15μU/mL in the winter .Through the analysis ,although the hTSH cut‐off of capillary blood spots that was used in the local center could be applied to dried venous blood spots ,but not ideal cut‐off value .Even‐tually ,the hTSH P99 9 .20μU /mL in the spring was identified as cut‐off value of dried venous blood spots for routine screening la‐boratory ,which had a sensitivity of 100 .00% ,a specificity of 36 .80% ,the diagnosis rate of 38 .96% ,and no missed phenomena .For the better service ,the hTSH value in the range of 9 .20-20 .00 μU/mL was defined as the boundary region ,and it was the danger region when the hTSH value was higher than 20 .00 μU/mL .Conclusion It is necessary to establish a reasonable hTSH cut‐off value to provide objective evidence for the use of dried venous blood spots of neonatal screening for congenital hypothyroidism ,ac‐cording to the boundary region and the danger region ,the recall chosen a different way ,the development of neonatal screening can be effectively promoted .

To explore the therapeutic effect of micro-traumatic surgery of loose seton and cutting seton by rubber bands in the treatment of high anorectal fistulae.Application of cutting seton (truss rubber bands) implemented the high part of fistulae and loose seton (ligation rubber band but non-fastened) for the low part of fistulae.133/136 patients undergoing micro-traumatic surgery were cured by one operation,2 cases had pseudo-healing and there was 1 recurrent case.And the curative rate was 97.8％.The microtraumatic surgery of loose seton and cutting seton by rubber bands in the treatment of high anorectal fistula has such multiple advantages as small incision,minor trauma,lesser pain,faster healing and a shorter course of treatment.And it may preserve the proper anal function and the integrity of anal skin.And its clinical efficacy is satisfactory.

BACKGROUND:Numerous studies have shown that local lumbar stenosis can cause immunological abnormalities and local chronic inflammation, which is the main cause of pain. At present, studies on inflammatory factors and lumbar spinal stenosis mainly focused on intervertebral discs, facet joint and ligamenta flava. No reports addressed the relationship between inflammatory factor in vein of lumbar spinal canal and lumbar spinal stenosis. OBJECTIVE:To analyze the correlation of serum interleukin-1αand tumor necrosis factor-αlevels with lumbar spinal stenosis. METHODS:A total of 51 patients with lumbar spinal stenosis or lumbar vertebral burst fracture, who underwent posterior lumbar decompression in the Department of Spine Surgery, Shanghai East Hospital, Tongji University in China from September 2011 to December 2013, were enrol ed in this study. Visual analogue scale score of low back pain and Oswestry disability index were evaluated before treatment. Peripheral vein blood and venous blood in the vertebral canal were col ected from patients with lumbar spinal stenosis or lumbar vertebral burst fracture. The concentrations of serum interleukin-1αand tumor necrosis factor-αwere determined using enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION:The concentration of interleukin-1αin degenerative lumbar stenosis group was significantly higher than that in the lumbar burst fracture group and peripheral veins (P<0.05). The more segments of lumbar spinal stenosis, the higher the venous serum interleukin-1αlevels were in the degenerative lumbar stenosis group, but the statistical difference was not significant. Linear correlation analysis results displayed that interleukin-1αlevels were positively associated with low back pain and disability scores in the degenerative lumbar stenosis group (r2=0.359 3, P<0.05;r2=0.526 4, P<0.05). These results indicated that the lumbar spinal venous inflammatory factors may be one of the reasons of low back pain and dysfunction in patients with degenerative lumbar spinal stenosis.

ObjectiveTo observe the lung injury of rats caused by PM2.5 induced imbalance of TH17/Treg immune system and the intervention effect of two different TCM treatments.Methods Wistar male rats were randomly divided into normal group, model group, TCM-treated group1 and TCM-treated group 2. PM2.5-induced lung injury model was established by airway instillation. Model group was given normal saline for gavage. TCM- treated group 1 and 2 were given Yupingfeng Powder and Guomin Decoction combined with Zhisou Powder for gavage. The pathological changes of bronchial and lung tissues, the contents of IL-8, IL-10, IL-17, NE, and MUC5AC in serum and BALF were compared, and the expressions of Foxp3 and IL-17 in lung tissue of each group were analyzed.Results Compared with normal group, the contents of IL-8, IL-17, NE and MUC5AC in serum and BALF of model group increased significantly, while IL-10 decreased significantly (P<0.05,P<0.01); the expression of IL-17 increased significantly and the expression of Foxp3 decreased significantly in lung tissue (P<0.01). Compared with model group, the contents of IL-8, IL-17, and NE decreased in TCM-treated group 1 and 2, while the content of BALF IL-10 increased significantly (P<0.05). The content of IL-10 in serum increased significantly in TCM-treated group 2 (P<0.05); the protein expression of IL-17 of lung issue decreased significantly, and the protein expression of Foxp3 increased significantly (P<0.01). The pathological changes were improved significantly.Conclusion PM2.5 can induce lung injury caused by the imbalance of TH17/Treg. Both two treatments can significantly improve the lung injury induced by PM2.5 and the imbalance of TH17/Tregs immune system.

Objective To prepare natural anti-keratin IgM monoclonal antibody. Methods Spleen cells of BALB/c mice raised in specific pathogen free conditions were directly fused with Sp2/0 myeloma cells. The hybridoma supernatants were tested by ELISA using pre-extracted keratin. The natural IgM obtained was further identified by immunochemistry and immunoblot methods. Results The cell fusion rate was about 60% without pre-immunization. About 14% supernatants reacted with the keratin antigen. Three hybridoma strains secreting natural IgM monoclonal antibody against keratin were obtained. The immunochemistry results showed that the natural anti-keratin IgM was able to bind to epidermis, sebaceous gland, hair follicule, and muscle tissues. Conclusion B lymphocytes in normal BALB/c mice spleen can produce natural antibody against kerain.

Objectives To investigate the binding of natural IgM to Staphylococcus aureus and its role in the phagocytosis of S. aureus by phagocytes, and to pave way for further study on the role and mechanism of natural IgM in defense of bacteria. Methods The binding of natural antikeratin IgM 3B4 to S. aureus was analyzed by ELISA and indirect immunoiluorescence. The role of 3B4 in the phagocytosis was analyzed by colony forming assay and flow cytometry (FCM). Results Both ELISA and indirect immunofluorescence proved the binding of natural IgM 3B4 to S. aureus. Colony forming assay found that the amount of colony forming units decreased significantly when 3B4 was added. The analysis of FCM showed that 3B4 augmented phagocytosis of 5. aureus by phagocytes. Conclusions Natural antikeratin IgM 3B4 can bind to S. aureus and regulate the phagocytosis of it, indicating that natural IgM may play some role in the defense against bacterial infection.

Objective To investigate UV- or heat-inactivated Epstein-Barr virus(EBV)stimulating the production of anti-keratin autoantibody(AK auto Ab)in cultured human umbilical cord blood B cells. Methods Mononuclear cells were isolated routinely from umbilical cord blood, in which monocytes, NK cells and cytotoxicity T cells were eliminated by L-leucine methyl ester method, and T cells were removed by sheep red blood cells(SRBCs)treated with 2-amino ethyl-isothiouronium bromide (AET). The purified B cells were treated with UV- or heat-inactivated EBV respectively and then cultured in complete IMDM. CD5, CD3, CD4 and CD8 cells were detected by flow cytometry. IgG and IgM of AK auto Ab were measured by ELISA in the supernatant which came from the B cells treated by UV-inactivated EBV or EBV-transformed B cells respectively. Results In UV-inactivated EBV group CD5+B cells accounted for 43% and 47% of all cells detected on the 14th and 28th day, respectively. No CD3, CD4 and CD8 cells were detected during this period. In UV-inactivated EBV group the AK auto Ab of IgG and IgM increased significantly on the 18th and 26th day, respectively (P 0.05). On the 40th day the AK auto Ab of IgG and IgM were significantly higher in EBV-transformed B cell group than those in UV-inactivated EBV group. Conclusions UV-inactivated EBV is able to induce AK auto Ab production but heat-inactivated EBV does not, which suggests that EBV protein might be the effective agent in inducing the production of AK auto Ab.

Objective To systematically evaluate the effects of prone position ventilation on newborn with respiratory distress syndrome.Methods We searched databases including PubMed,EMBASE,Cochrane Library,Web of Science,CBM,CNKI,Wanfang and VIP.We included all randomized controlled trials and randomized crossover trials of neonatal respiratory distress syndrome.Studies were selected according to inclusion and exclusion criteria,extracting data and assessing quality.Then RevMan 5.3 software was used to analyze the data.Results Ten studies included seven randomized controlled trials and three randomized crossover trials,and 500 patients were included.The results of meta-analysis showed that in prone position ventilation group SaO2[MD=2.41,95％CI(0.87,3.95),P=0.002],PaO2[MD=5.20,95％CI(3.04,7.36),P＜0.001],Pa2//FiO2[MD=24.40,95％CI(8.35,40.44),P=0.003],the risk of pneumothorax [RR =0.10,95％ CI (0.01,0.76),P=0.03] and intracranial hemorrhage [RR =0.41,95％ CI (0.20,0.83),P=0.01]and duration of mechanical ventilation [MD=-23.22,95％CI(-38.30,-8.14),P=0.003] compared with supine positiongroup had significantly statistical difference;however there was no significant difference in ventilator-associated pneumonia[RR=0.72,95％CI(0.48,1.09),P=0.12] between two groups.Conclusion Prone position ventilation is effective to improve SaO2,PaO2 and PaO2/FiO2,shorten duration of mechanical ventilation and reduce the risk of pneumothorax and intracranial hemorrhage in newborn with respiratory distress syndrome.While it could not effectively reduce incidence of ventilator-associated pneumonia.Due to the limitations of the quality of studies included,multi-centered randomized controlled trials with large sample size are needed in the future to evaluate the effects of prone position ventilation on neonatal respiratory distress syndrome.

OBJECTIVETo study the expression of EpCAM and E-cadherin in papillary thyroid carcinoma and to analyze its correlation with various clinicopathologic parameters.

METHODSImmunohistochemical study for EpCAM and E-cadherin was carried out in 91 cases of papillary thyroid carcinoma. Twenty-four cases of papillary hyperplasia of thyroid were used as controls.

RESULTSIn all of the 24 cases of papillary hyperplasia, EpCAM was located on the cell membrane, while in the 91 cases of papillary thyroid carcinoma studied, EpCAM was located within the cytoplasm, with 36.3% (33/91) showing nuclear localization as well. In all the papillary hyperplasia cases studied, E-cadherin showed membranous expression. E-cadherin expression was reduced in 84.6% (77/91) of papillary thyroid carcinoma, as compared with the surrounding native thyroid parenchyma. Amongst the 33 cases of papillary thyroid carcinoma which showed nuclear localization of EpCAM, 30 cases also showed reduced E-cadherin expression. There was a positive correlation between nuclear expression of EpCAM and loss of E-cadherin expression (P = 0.000; Spearman correlation coefficient = 0.857). Nuclear expression of EpCAM correlated with follicular variant of papillary thyroid carcinoma and presence of extrathyroidal extension ( P = 0.037 and 0.033, respectively). Loss of E-cadherin expression correlated with age of patients and presence of lymph node metastasis (P = 0.018 and 0.010, respectively).

CONCLUSIONSE-cadherin expression is reduced in papillary thyroid carcinoma, as compared with native thyroid parenchyma and papillary hyperplasia. Papillary thyroid carcinoma shows loss of EpCAM membranous expression and increased cytoplasmic/nuclear accumulation. Detection of these two markers may provide a valuable reference in defining the biologic behaviors of papillary thyroid carcinoma, including extrathyroidal extension and lymph node metastasis.

Antigens, Neoplasm ; metabolism ; Cadherins ; metabolism ; Carcinoma, Papillary ; metabolism ; secondary ; Cell Adhesion Molecules ; metabolism ; Cell Membrane ; metabolism ; Cytoplasm ; metabolism ; Epithelial Cell Adhesion Molecule ; Humans ; Lymphatic Metastasis ; Neoplasm Proteins ; metabolism ; Thyroid Neoplasms ; metabolism ; pathology