Objective To observe the effects of substance P (SP), NK1 receptor antagonist and nitric oxide synthase (NOS) inhibitors on the secretion of nitric oxide (NO) and expression of inducible NOS (iNOS) by immortalized human keratinocyte line HaCaT. Methods The NO level in supernatant of cultured HaCaT cells was measured by nitrate reductase assay after treatment with different concentrations (10-9 to 10-6 mol/L) of SP, or the combination of SP (10-8 mol/L) and spantide (3×10-7 mol/L), aminoguanidine (10-7 mol/L), 7-nitroindazole (10-6 mol/L) or L-NAME (10-5 mol/L) for various durations. Reverse transcription-PCR was performed to measure the expression of iNOS mRNA in HaCaT cells incubated with SP of 10-8 mol/L for 1, 24 and 48 hours. Results The SP of 10-9 to 10-6 mol/L significantly induced the production of NO by HaCaT cells, and the highest level of NO was observed in HaCaT cells treated with SP of 10-8 mol/L. The synthesis of NO by HaCaT cells induced by SP was inhibited by Spantide of 3 × 10~7 mol/L at all time points (30 minutes, 1, 3, 6, 12, 24 hours, all P< 0.01), by L-NAME of 10-5 mol/L at 3 time points (30 minutes, 1, 24 hours) and by 7-nitroin-dazole of 10-6 mol/L at 2 time points (30 minutes, 1 hour, both P< 0.05), but not by aminoguanidine of 10-7 mol/L at any time point (all P > 0.05). After treatment with SP of 10-8 mol/L, the relative mRNA expression of iNOS was 0.199 ± 0.018 and 0.516 ± 0.030 at 24 and 48 hours, respectively, and there was a statistical difference between the two time points (P < 0.01). Conclusions SP can reinforce the secretion of NO by HaCaT cells via NK1 receptor activation, but iNOS is unlikely to be the primary origin of NO secreted by HaCaT cells induced by SP.

Objective To evaluate multi- slice spiral CT angiography (MSCTA) in diagnosing bronchial-pulmonary shunts (BPS). Methods The materials of MSCTA images in 31 patients with DSA-confirmed BPS were retrospectively analyzed. Taking DSA imaging as the reference standard, the consistency in diagnosing BPS at different levels between MSCTA and DSA was compared. Results On MSCTA using aorta-density trigger scanning technique BPS was manifested as wrong-time enhancement of pulmonary artery branches. Wilcoxon test indicated that MSCTA had a high concordance with DSA in detecting BPS occurring at sub-segmental (or above) pulmonary artery branches (Z = -1.854, P > 0.05), but MSCTA detection of BPS occurring at peripheral pulmonary arteries was obviously poorer than DSA (Z = -3.923, P < 0.05). MSCTA confirmed the diagnosis of BPS in 11 cases with a sensitive rate of 35.5% (11/31), and no differences in revealing the number and the shunt levels of BPS existed between DSA and MSCTA. The number of abnormal systemic arteries demonstrated by MSCTA was quite the same as the number detected by DSA. No false positive case was seen. Conclusion MSCTA is very effective in diagnosing high-flow BPS locating at sub-segmental (or above) pulmonary artery branches as well as in revealing abnormal bronchus-related systemic arteries, which is very helpful in preoperatively evaluating the severity of the BP shunting and the supplying arteries to be obstructed.

Objective To evaluate the ethical management status quo of Capital Medical Research Fund in 2007.Methods Cross-sectional study was applied to analyze the ethics management and ethics self-evaluation in applications.Results There were 652 applications,covering with 14 districts and 2 counties in Beijing,from 128 Hospitals.Applicants have some knowledge of the ethical issues in clinical research accounted for 88.7％ ; informed consent of subjects considered in 72.5％ ;the potential risk and protection involved in application accounted for 62.0％; personal privacy protection of subject accounted for 49.4％.Unfortunately,the benefit vs risk assessment was only involved in 28.7％ applications.Applicants from general hospitals had more ethical issue knowledge (90.2％) than those from community hospitals (84.6％).Applicants from university hospitals had more ethical issue knowledge (93.9％) than those from military hospitals (80.2％).Applicants aged 55 0r over had less ethical issue knowledge (70.0％) than other applicants (89.2％).The applicants to study on descriptive research and etiological research had less ethical issue knowledge than others.578 applications filled out the approval from ethics committee or research management department.62.6％ of the written were acceptable.Conclusion The applicants for clinical research had preliminary understanding for ethical issues in Beijing in 2007.Hospital ethics committees or research management departments had conducted clinical research ethical review applications for funds management.It has taken place the external conditions to carry out the ethical management in clinical research fund management.

AIM: To study the determination of 5 kinds of cholic acid content. METHODS: Using single standard substitue (SSS), we assayed 5 kinds of cholic acid, such as cholic acid, deoxycholic acid, hyodexycholic acid, chenodeoxychoilc acid and ursodeoxycholic acid by HPLC/ELSD. and studied the influence of the additive substitutes (such as rabbit Serum, urine and Qingkailing extract) on the measure values. RESULTS: By using any one of the 5 kinds of cholic acid standard, the content in all the 5 kinds of cholic acid could be determined. CONCLUSION: Simply as well as credibly, the SSS method could be used in quality analysis, especially in complex matrix. Also, it proved a feasible way in quality control and research of fingerprint chromatography of TCM.

Background Selective laser trabeculoplasty (SLT) can increase the outflow of aqueous humor and reduce the intraocular pressure of patients with open angle glaucoma,but its mechanism is unknown.To investigate the mechanism of SLT would improve the therapeutic effect of SLT.The aqueous outflow resistance in trabecular meshwork was affected by the extracellular matrix (ECM).Matrix metalloproteinase-3 (MMP-3) and MMP-9 were closely related to ECM degradation in trabecular meshwork.Objective This study was to observe the effects of SLT on the expressions MMP-3 and-9 in human trabecular ceils in vitro.Methods Immortallized human trabecular cells were cultured with pigment particles mixed suspension for 16 hours and incubated overnight.Then the cells were irradiated with Q switch frequency doubling 532 nm Nd:YAG laser to establish SLT-effective cells with the energy of 0.2 mJ,spot diameter of 400 μm and pulse duration of 3 ns.The expressions of MMP-3 mRNA and MMP-9 mRNA in the cells were detected by fluorescence quantitative real time PCR before and 1 hour and 4,8,12,28 hours after exposure of laser.The concentrations of MMP-3 and MMP-9 in the medium were assayed using ELISA 1 hour and 4,8,12,28 hours after exposure of laser and compared between the non-irradiation group and the irradiation group.Results The relative expressing levels of MMP-3 mRNA were 1.00,1.32±0.12,2.08±0.05,2.34±0.04,2.77± 0.05 and 2.49±0.27 in the non-irradiation group and irradiation for 1 hour and 4,8,12,28 hours after exposure of laser SLT,showing a significant difference among the groups (F =15.966,P=0.007),and relative expressing levels of MMP-3 mRNA were significantly higher in various time points after laser irradiation than those of the non-irradiation group (all at P＜0.05).The relative expressing levels of MMP-9 mRNA were 1.00,0.91 ±0.10,1.27 ± 0.07,1.46±0.07,1.69±0.09 and 0.87±0.09 in the non-irradiation group and irradiation for 1 hour and 4,8,12,28 hours after exposure of SLT,which was considerably different among the groups (F =30.526,P =0.005),and significant increased values were seen in the 4,8 and 12 hours after irradiation compared with the non-irradiation group (all at P＜0.05),with highest expression in the irradiation for 12-hour group.The concentrations of MMP-3 and MMP-9 proteins in medium were significantly increased in various time points after laser exposure in comparison with the control group (all at P＜0.05).Conclusions The expressions of MMP-3 and MMP-9 in human trabercuolar cells upregulate and the secretion ability of human trabecular cells to MMP-3 and MMP-9 proteins improves in early stage of SLT in vitro.However,these procedures gradually diminish with the lapse of time.

Background Selective laser trabeculoplasty (SLT) plays lowing-intraocular pressure (IOP)effect by irradiating pigmented trabecular cells selectively using 532 nm Nd:YAG laser.However,its affect pattern is not fully known.Objective This study was to establish a human trabecular cells (HTCs) model of SLT effect,and to observe the morphological changes of HTCs after they were exposed to different energies of SLT.Methods Immortalized HTCs were incubated in DMEM/F12 with pigmental granules for 16 hours (overnight) to prepare pigmented trabecular cells.The cells were irradiated with modified laser lens with the spot of 400 μm and emitting duration of 3 ns.The energy scale of 0.2 mJ was set for standardized energy and 0.1 mJ was used as the low energy.The morphology and ultrastructure of the cells were examined under the light microscope and transmission electron microscope 1 hour,4,8,12,24 hours after irradiation.Trypan blue staining and TUNEL fluorescence staining were used to evaluate the death and apoptosis of the cells after laser irradiation.Results The brown pigment particles were seen in cytoplasm around nuclei of trabecular cells after cultured for 16 hours.After laser irradiation,there was no obvious change in the shape of nonpigmented trabcular cells,but a 400 μm spot was found in the pigmented trabecular cells under the light microscope,and the pigmented particles released as the cell disruption.After 0.2 mJ laser irradiation,cell loss zone was seen at the center of the laser spot,and the positive cells for trypan blue and TUNEL staining were found;while the abnormal manifestations were slight after the 0.1 mJ irradiation.With the lapse of the time,the number of positive staining cells was gradually decreased.Twenty-four hours after laser irradiation,proliferative trabecular cells emerged and migrated to the center area of laser spot.Statistical analysis showed that the numbers of the positive cells for trypan blue and TUNEL staining were significantly reduced in the lower energy group in comparison with the standardized energy group at various time points (all at P＜0.001),and those of both energy groups were gradually reduced with lapse of time with the significant differences between any two time points (all at P＜0.01).Conclusions An in vitro laser effect model of HTCs can be established by exposing pigmented HTCs to SLT laser.After exposed to SLT,the gasification,necrosis-like death,apoptosis-like change appear at the laser center.The laser destroyed cells can be replaced by proliferative cells with the lapse of time.Low-energy laser irradiation cause a similar morphological change of the cells to high-energy irradiation,but the number of abnormal cells is much less.

Objective To improve the recognition of pituitary hyperplasia secondary to primary hypothyroidism by analyzing clinical and imaging data. Methods The clinical features, hormone data,imaging findings, and treatment were reviewed in 32 patients with pituitary hyperplasia secondary to primary hypothyroidism in Huashan Hospital from 1999 to 2008. Results Thirty-two patients, most juvenile,presented clinical and imaging features suggestive of functional primary pituitary adenoma. The dose of levothyroxine was increased to maintain the thyrotropin concentration at normal values. Following adequate thyroxine replacement, pituitary hyperplasia regressed on average within 6 months. Conclusions Pituitary hyperplasia secondary to primary hypothyroidism seems to be quite prevalent in children and adolescents.Complete regression will be achieved with thyroxine replacement therapy.

Objective To evaluate the predictive value of N-terminal-pro-brain natriuretic peptide (NT-proBNP) in weaning patients from mechanical ventilation (MV).Methods Data of 42 patients supported with MV in intensive care unit (ICU) admitted to the Rui Jin Hospital from January through December in 2014 were retrospectively analyzed,and the causes for MV were recorded.According to the outcomes of weaning from MV after 48 hours,the patients were divided into two groups namely success group and failure group.Comparisons of fluid balance in 72 hours before spontaneous breathing trial (SBT),and comparisons of NT-proBNP1 levels at admission,NT-proBNP2 levels before SBT,NT-proBNP3 levels after 48 hours after SBT between two groups were carried out.And the receiver operating characteristic (ROC) curve for predicting weaning rate was plotted to find the optimal cut-off point of NT-proBNP2.Results In the total of 42 patients,there were 27 cases in success group and 15 cases in failure group.There were not statistically differences of NT-proBNP1 levels between success group and failure group (P =0.121).However,the NT-proBNP2 levels and NT-proBNP3 levels in failure group were significantly higher than those in success group (P =0.01,0.003).The area under curve (AUC) of the ROC curve of NT-proBNP2 levels to predict the failure of weaning was 0.862 (95％ CI:0.753-0.971).When the optimal cut-off point of NT-proBNP2 was 715.5 pg/mL,the sensitivity and specificity were 93.3％ and 74.1％,respectively.Conclusion The NT-proBNP2 levels before SBT have predictive value in weaning rate,and it can be used as one of the screening indicators for weaning.

AIM: To investigate the protein patterns of bone marrow mesenchymal stem cells(MSCs) induced by 5-azacytidine and Shuanglong Formula(Radix et Rhizoma Ginseng,Radix et Rhizoma Salviae Miltiorrhizae) and molecular mechanism of induced differentiation was discussed at the level of proteome. METHODS: MSCs extracted from Chinese miniswine were induced by 5-azacytidine and Shuanglong Formula.Proteins were separated by two-dimensional gel electrophoresis,and were detected by silver staining method.The image was analyzed by PDQuest software,and the intensities of spots in the gels were used to quantify the expressions of proteins.Proteins of interest were chosen for in-gel digestion and MALDI-TOF-MS analysis,and then further identified by peptide mass fingerprinting(PMF). RESULTS: 16 altered proteins were identified by PMF and their regulation might be caused by the addition of medicate sera containing the active components of Shuanglong Formula.The effects of medicated sera containing Shuanglong Formula on the induced-differentiation could be discussed from the regulated expression of functional proteins; CONCLUSION: Shuanglong Formula may promote the 5-azacytidine-induced differentiation of bone marrow MSCs in vitro,and our study indicate that proteomics can be used for the analysis of molecular mechanism for different cell process.

Objective To raise the level of clinical diagnosis for the patients with pituitary stalk interruption syndrome (PSIS). Methods Five patients (4 males) with PSIS were retrospectively analyzed with respects to the clinical features, endocrine status and image characteristics. Magnetic resonance imaging (MRI) scan was performed at hypothalamic-pituitary region. Results The clinical manifestations of all patients consisted of growth retardation and delayed puberty without polyuria. Deficiency of multiple anterior pituitary hormones was revealed in all patients by the evaluation of endocrine status. The features of MRI included a lack of visible pituitary stalk, absence of posterior lobe hypersignal in the sella turcica and a hyperintense spot in the region of the thalamus opticus. Conclusion The clinical characteristics of patients with PSIS are growth retardation and delayed puberty. The evaluation of anterior pituitary function is necessary. The detection of an anatomical abnormality around hypothalamic-pituitary region by MRI is important diagnostic evidence.