Objective:To establish a method for the detection of bacterial endotoxins in tertiary butyl alcohol. Methods:The in-terference experiment and bacterial endotoxins test for 3 batches of the products from 2 companies were carried out with tachypleus ame-bocyte lysate. Results:There was no interference in the bacterial endotoxins test. The limit of bacterial endotoxins was 0. 25 EU/ml. Conclusion:The established method of bacterial endotoxins test is feasible for tertiary butyl alcohol.

Objective To explore the role of MAPK and NF-?B in the increase of Toll-like receptor 4(TLR4) expression induced by lipopolysaccharide(LPS) in pulmonary micro-vascular endothelial cells(PMVECs) of mouse.Methods PMVECs were isolated,cultured and exposed to the different concentration LPS at(2 h),and(100 ?g/L) LPS in different time.The concentration of TNF-? in culture supernatant was detected using ELISA.(Using) PD98059 inhibitor for ERK,SB203580 inhibitor for P38 MAPK and PDTC inhibitor for NF-?B pretreated(PMVECs).RT-PCR was used to detect the levels of TLR4 and TNF-? mRNA.Western-blot was used to detect the protein expression of nuclear factor-kappa B(NF-?B) P65 subunit and ERK / P38 MAPK in nuclear extract after LPS exposure for 6 h respectively.Results With the treatment of 50～(500 ng /mL) LPS for(2 h) or(100 ?g /L)(LPS exposure) in different time,the concentration of TNF-?in culture supernatant of PMVECs was increased by a time and dosage dependent style(P

To screen potential human soluble epoxide hydrolase (hsEH) inhibitors, a high-throughput screening model in 384-well microplate with total volume of 50 microL was established. Recombinant hsEH was cloned and expressed in E. coli. and its specific substrate PHOME was synthesized. The HTS model was based on fluorescence analysis with enhanced sensitivity and specificity (Z' = 0.65). A total of 47 360 samples (including 25 040 compounds and 22 320 natural products) were screened, of which 950 samples with inhibition greater than 80% were selected for further rescreening. Finally, two compounds with high inhibitory activity were identified, whose IC50 value were 8.56 and 4.31 micromol x L(-1), separately. The results indicated that the method was stable, sensitive, reproducible and also suitable for high-throughput screening.

BACKGROUND:There are two types of epithelial stem cells in the ocular surface tissue:corneal epithelial stem cells and conjunctival epithelial stem cells. The corneal epithelial stem cells play an important role in renewal of corneal epithelial cells and maintenance of corneal transparency.
OBJECTIVE:To study the location of corneal epithelial stem cells using laser in vivo confocal microscopy and immunofluorescent staining.
METHODS:Patients with unilateral limbal stem celldeficiency who went to Henan Eye Institute from September 2009 to September 2012 were enrol ed in this study. Bilateral eyes were scanned by laser in vivo confocal microscopy, and the healthy eye was imaged as a control. The central cornea and limbus were scanned and images were recorded for statistical analysis. The eye bal s were obtained from Henan Eye Bank, China. Central cornea and limbus were dissected and embedded in the OCT compound for frozen section and the proper thickness of the section was 5-7μm. Immunofluorescent staining was used to detect the expression of p63, ABCG2, K3 and Connexin 43 in the epithelial layers of central cornea and limbus.
RESULTS AND CONCLUSION:Twenty-four patients diagnosed with unilateral limbal stem celldeficiency were recruited. Under confocal microscopy, in the affected eyes, the typical morphology of conjunctival epithelial cells and goblet cells was detected instead of corneal epithelial cells;in the limbus, a great amount of fiber scarring tissue was detected instead of Vogt palisade, rete pegs and pigment cells. Immunofluorescent staining showed the expression of p63, ABCG2 was mainly in the basal layer of limbal epithelium, especial y in the outer and middle parts, but the expression of p63 and ABCG2 was not detected in the epithelial celllayers of central cornea. K3 and Connexin43 were not expressed in suprabasal layers of limbal epithelium, but in central cornea, they were expressed highly in the whole epithelial celllayers. Laser in vivo confocal microscopy and immunofluorescent staining showed the corneal epithelial stem cells were located in the basal layer of outer and middle limbal epithelium, mainly in Vogt palisade and rete pegs.

Objective To investigate the expression level of CD226 mRNA in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE) and explore the relation between the gene expression and disease activity, and the relation between the gene expression and Gly307Ser polymorphism of CD226 was also examined. Methods CD226 gene was measured with real-time polymerase chain reaction (qRT- PCR) in PBMCs. The expression levels of CD226 gene in PBMCs were compared between 90 SLE patients and 30 healthy individuals. One-way ANOVA and pearson correlation were used for statistical analysis. Results The expression level of CD226 in the PBMCs of SLE patients (6.8±1.1) was significantly decreased compared to healthy individuals (26.5±6.7) (P＜0.01), while there was no association between mRNA level and genotype (P＞0.05). No correlation between ESR, CRP, ANA, SLEDAI scores, C3 and the expression level of CD226 gene was discovered. Conclusion In Hubei Chinese Han population, CD226-Gly307Ser locus is associated with the development of SLE, while T allele does not impact the expression of CD226 gene, thus the role of CD226 gene in autoimmune diseases should be explored in the future.

Objective To explore the clinical characteristics of treatment?resistant depression ( TRD) and of its relevance with thyroid hormones. Methods 43 patients with TRD and 48 patients with non?TRD were as?sessed with 17?item Hamilton depression scale ( HAMD?17) and Hamilton anxiety scale ( HAMA) . The serum lev?el of thyroid?stimulating hormone ( TSH) ,total triodothgronine ( TT3) ,total thyroxine ( TT4) ,free triodothgronine ( FT3) and free thyroxine ( FT4) were determined by radioimmunoassay. χ2 test and t test were used for statistic a?nalysis. The quantitative relation of FT3 level with TRD was analyzed and the value of FT3 level in TRD diagnosis was evaluated by ROC curve.Results Compared with non?TRD patients,the TRD patients showed a younger dis?ease onset age ((16.98±2.25)years vs (23.50±3.38)years) and a longer disease course ((10.14±6.47)years vs (5.48±4.15)years) for total disease course;(60.35±23.64)months vs (5.00±3.40)months for current disease course),and had shorter education years ((8.53±1.72)years vs (11.04±2.07)years) and higher HAMD total scores (19.09±1.59 vs 15.69±2.38) and behavior retarding factor scores (8.72±0.98 vs 4.98±1.63). In addition, the FT3 level of TRD patients was lower than that of non?TRD patients ((3.92±0.15)pmol/L vs (4.16±0.20) pmol/L).All the above differences between two groups were statistically significant (P<0.05). The logistic regres?sion analysis showed that the risk of suffering TRD increased by 1. 006?fold when FT3 level reduced 0. 1 pmol/L (OR=2.006,95%CI=(1.501,2.681). The area under ROC curve was 0.821 (P<0.001) with its 95% confidence interval of (0.734,0.907). Conclusion Compared with non?TRD patients,TRD patients have a longer disease course,a younger disease onset age, a lower education level, higher HAMD total scores, more severe retardation symptoms,and a lower FT3 level. The serum FT3 level has a high reference value in diagnosis of TRD.

Post-marketing evaluation is a process which evaluate the risks and benefits of drug clinical application comprehensively and systematically, scientific and systematic results of post-marketing evaluation not only can provide data support for clinical application of traditional Chinese medicine, but also can be a reliable basis for the supervision department to develop risk control measures. With the increasing demands for treatment and prevention of disease, traditional Chinese medicine has been widely used, and security issues are also exposed. How to find risk signal of traditional Chinese medicine in the early stages, carry out targeted evaluation work and control risk timely have become challenges in the development of traditional Chinese medicine industry.
Drug Evaluation ; methods ; Drugs, Chinese Herbal ; Humans ; Medicine, Chinese Traditional

Objective To investigate whether paclitaxel can efficiently induce the apoptosis of ovarian cell HO-8910,and to study the relationship between the apoptosis of cells and the cell cycle.Methods With the treatment of paclitaxel with different concentrations and different time,the morphologic change of HO-8910 ovarian cells was observed using fluorescence microscopy and transmission electron microscopy(TEM),and the apoptosis of cells and the changes of cell cycle were determined by flow cytometry(FCM). Results The typical changes of HO-8910 cell apoptosis were observed by TEM and Fluorescence microscopy.With the treatment of paclitaxel,the HO-8910 ovarian cells were firstly arrested in G_2/M phase,and the typical ultrastructural changes of apoptosis were appeared only after the cells were apparently arrested in G_2/M phase.Conclusion Paclitaxel can induce the apoptosis of HO-8910 cells and the apoptosis is associated with the blockage of G_2/M phase in cell cycle.

Objective To explore the features of sustained attention in the patients with social anxiety disorder and their first degree relatives.Methods 60 cases of normal controls,58 cases of patients with social anxiety disorder and 68 cases of their first degree relatives were performed with continuous performance test.Results In reaction time task(a simple performance),all of CPT parameters in patient group (number of cancel (5.2±6.9),number of omission(6.7± 13.6),reaction time(421.0± 112.0) ms,variance coefficient(47.3±38.1) ％)were higher than those in the first degree relatives group(number of cancel (2.9± 3.3),number of omission (1.5±2.1),reaction time (342.0±47.0) ms,variance coefficient(28.9± 10.1) ％) and controlled group (number of cancel (2.8 ± 2.9),number of omission (1.3 ± 0.7),reaction time (329.0 ± 53.0) ms,variance coefficient (27.8± 9.7) ％),the differences were significant(P＜0.01),while all of CPT parameters in the first degree relatives were similar to those in controlled group(P＞0.05).In X task (a complex performance),the results of CPT in the first degree relatives were between the patients and the normal controls,and the differences were statistically significant(P＜0.05).Conclusion The anxiety disorder patients showed no sustained attention impairment in simple performance while in a complex performance there were some defects of sustained attention functions in the patients with social anxiety disorder and their first degree relatives.

ObjectiveTo study the effect of adaptive group psychological training on mental quality and emotion of soldiers.MethodsA total of 48 soldiers ( experiment group) received group psychological training and read a textbook,Mental Quality Training for soldiers,for 3 months,and the other 45 soldiers served as control group.Armyman mental quality inventory (AMQI),self-rating depression scale (SDS) and state-trait anxiety inventory(STAI) were employed to evaluate the mental quality and emotion of soldiers before and after training.All data were analyzed by paired-sample t test.Results ①As compared with pre-training,the AMQI scores of training group increased significantly in bravery ( 71.50 ± 9.59 ),self-confidence ( 68.24 ± 8.36 ),aptitude ( 74.54 ± 10.81 ),loyalty(75.29 ± 8.08 ) and general score ( 359.07 ± 20.30 ) (P ＜ 0.05,P ＜ 0.01 ) ; and no significant difference were found in those of control group(P ＞ 0.05 ).②When compared with pre-training,the scores of SDS (39.00 ±8.38),SAI(40.14 ±9.92),TAI(41.16 ±7.47) and STAI(81.30 ± 12.52) of training group decreased significantly (P ＜ 0.05,P＜ 0.01 ) ; and no significant difference was found in those of control group(P ＞0.05 ).ConclusionGroup psychological training can improve mental quality of soldiers,which is helpful to relax anxiety and depression of soldiers.