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.

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 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.

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.

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.

The adverse reaction monitoring is important in warning the risks of traditional Chinese medicines at an early stage, finding potential quality problems and ensuring the safe clinical medication. In the study, efforts were made to investigate the risk signal mining techniques in line with the characteristics of traditional Chinese medicines, particularly the complexity in component, processing, compatibility, preparation and clinical medication, find early risk signals of traditional Chinese medicines and establish a traditional Chinese medicine safety evaluation system based on adverse reaction risk signals, in order to improve the target studies on traditional Chinese medicine safety, effective and timely control risks and solve the existing frequent safety issue in traditional Chinese medicines.
China ; epidemiology ; Drug Evaluation ; Drug-Related Side Effects and Adverse Reactions ; epidemiology ; etiology ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Humans ; Product Surveillance, Postmarketing

Traditional Chinese medicine Polygoni Multiflori Radix is dried roots of Polygonaceae Polygortum multiflorum Thunb. Its clinical application records were first discovered in literatures of the Tang dynasty. The origins, efficacy, toxicity, processing and taboos of Polygoni Multiflori Radix have been discussed in many ancient herb literatures. In recent years, with the increase in the public awareness in health, Polygoni Multiflori Radix admits preparations have been more widely applied in the treatment and prevention of diseases. However, there have been more and more reports about Polygoni Multiflori Radix induced liver injury, the safety of Polygoni Multiflori Radix has increasingly attracted attention of the society. In this paper, the authors summarized and analyzed the toxicity and medication risk factors of Polygoni Multiflori Radix recorded in ancient herb literatures, and proposed that more attention shall be given to the effect of the planting and processing methods on the components and toxicity of Polygoni Multiflori Radix in safety studies, which provides clues for the further studies.
China ; Drugs, Chinese Herbal ; adverse effects ; history ; History, Ancient ; Humans ; Medicine in Literature ; Medicine, Chinese Traditional ; history ; Polygonum ; adverse effects ; Risk Assessment

Objective To investigate the relationship between the helper T cell 17 (TH 17)/ Regulatory T cells (Treg cells) balance in peripheral blood with acute graft-versus-host reaction (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT),as well as the impact of anti-thymocyte immunoglobulin (ATG) on helper T cells in peripheral blood.Method Seventyeight hematologic patients underwent allo-HSCT,conditioning with or without ATG.Ten healthy volunteers severed as a control group.The helper T and regulatory T cells in peripheral blood were detected by flow cytometry.Enzyme-linked irnmunosorbent assay (ELISA) was used to detect serum concentrations of interleukin(IL)-17,IL-21,IL22,IL23,γ interferon (IFN-γ),and transforming growth factor β1 (TGF-β1).Result The percentage of Treg cells,TH17 cells and ratio of TH17/Treg cells in patients without aGVHD showed no significant difference from the healthy controls (P＞ 0.05).As compared with control group and non aGVHD group,the ratio of Treg cells was increased,the percentage of TH 17 cells,and TH 17/Treg cells were significantly increased in 1-2-degree aGVHD group (P＜0.01).With increased degree of aGVHD,the difference as above was more significant in 3-4-degree aGVHD recipients (P＜0.01).In aGVHD group,the IL-17,IL-23,IL-21 and IFN-γ concentrations were higher than the healthy group (P＜0.01) and non-aGVHD group (P＜0.05).Serum TGF-β1 level in aGVHD group was significantly decreased as compared with healthy group and non-GVHD group (P＜0.05),while IL-22 concentrations showed no statistically significant difference among three groups (P＞0.05).In anti-thymocyte immunoglobulin (ATG) pretreatment group,the absolute count of peripheral blood lymphocytes was less than in healthy control group (P＜0.01).In ATG group,the absolute counts of TH1 cells,TH17 cells,CD3+ CD4+ cells and non-TH1/17 cells were less than in non-ATG group (P =0.0000),while the absolute counts of lymphocytes,CD3+ CD4-cells,and TH 1/17 cells were less than in non-ATG group,but there was significant difference (P＞0.05).Conclusion The balance of TH 17/Treg cells and related cytokines were closely associated with aGVHD after allo-HSCT,and ATG influences the reconstruction of TH 17 and Th1 cells at early stage.