Objective To explore the effects of homocysteine (Hcy) on neural stem cell (NSC) proliferation and the mRNA expression level of Notch1 and Hes1 related Notch signaling pathway from neonatal rats in vitro. Methods NSCs from neonatal rats were cultured by serum-free culture method in vitro. Cells were divided into four groups: control group (Hcy-C), low dose Hcy (Hcy-L) group, middle dose Hcy (Hcy-M) group and high dose Hcy (Hcy-H) group. NSCs were iden- tified by immunofluorescent staining using the antibodies against Nestin, β-tubulin Ⅲ and GFAP. The proliferation ability of NSCs was detected by MTT. The mRNA expressions of Notch1 and Hes1 were detected by Real-time PCR. Results In the serum free suspension medium, neurospheres that consisted of a great number of nestin-positive cells were found. β-tu- bulin Ⅲ positive neurons and GFAP positive astrocytes were detected by immunofluorescence staining on the 6 th day of cell induction. MTT assay showed that the cell viability was significantly lower in three Hcy treatment groups than those of con- trol group (P ＜ 0.05). And the effect of concentration-dependent was observed. The results of RT-PCR showed that mRNA expression of Hes1 was significantly lower in three Hcy treatment groups than that in control group (P ＜ 0.05). The mRNA ex- pression of Notch1 was significantly lower in Hcy-H group than that of other three groups (P ＜ 0.05). The mRNA expression of Notch1 was significantly lower in Hcy-M group than that of Hcy-L group and control group (P ＜ 0.05). Conclusion Hcy could inhibit the proliferation of NSCs by down-regulating mRNA expression levels of Notch1 and Hes1 genes related to Notch signal pathway.

The core-shell nanopaticles of Au@polyvinyl-pyrrolidone ( PVP) with uniform size and controllabe shell-thickness were prepared by hydrothermal method. The core-shell nanoparticles could be assembled to be the monolayer array on Si substrate relying on the dispersion of core-shell nanoparticles arising from PVP shell. The malachite green ( MG ) absorbed by H-bond could be detected on the array under the electromagnetic enhancement of inner-core Au nanoparticles. Under the conditions of the optimum shell-thickness of Au@PVP and the appropriate absorbed time of MG, the detection of MG could be realized in the linear range from 1 × 10-10 mol/L to 1 × 10-5 mol/L with the correlation coefficient ( R2 ) of 0. 98. The detection limit was 10-12 mol/L. This method was applied to the determination of MG in tilapia fish fillets of Xiagang market. No MG was found in this real sample. The spiked recoveries of the sample ranged from 70. 8% to 126. 0%. This method is simple and accurate, and can be used for detection of MG in the fish.

Prostate cancer is one of the most common malignant tumors in men and related studies have achieved great breakthrough in recent years.But because of the lack of effective in vivo animal models, the process to translate basic research into clinical application has been severely hampered.Patient derived prostate tumor xenograft ( PDPTX) model is an ideal animal model in which freshly isolated tumor tissues from patients were inoculated into immunodeficient mice.This model can duplicate the heterogeneity of primary tumor in a better way and keep the tumor complexity at molecular, genetic and pathological levels.Particularly, the PDPTX model, in which the isolated tumor tissue is inoculated under the renal capsule, is even better, because it solves the clrawbacks of traditional subcutaneous inoculation model.In traditional mod-els, the success rate is low, it’s not easy for lower grade tumor to form xenograft, and it’s not easy to reconstruct metasta-sis, etc.PDPTX provides a more ideal in vivo model for prostate cancer studies.It has irreplaceable advantages, especially in target therapy, new drug screening and individualized tumor treatment.

ObjectiveTo assess the differences of short- and long-term postintervention status on ocular and systemic symptoms for patients with ocular myasthenia gravis (OMG) after pyridostigmine bromide, corticosteroid, thymectomy, or thymectomy-corticosteroid combination therapy ( combination ).MethodsThis retrospective plus prospective study included 180 OMG patients, whose age of onset ≥ 15 years, treated non-randomly with above therapies separately: thymectomy group (60 cases ), corticosteroid group (39 cases), combination group ( 31 cases ), symptomatic group ( 50 cases ). Postintervention status complying with Myasthenia Gravis Foundation of America (MGFA)complete stable remission ,pharmacologic remission, or minimal manifestations was considered as desirable response, which was used as statistical indicator. Results ①Corticosteroid group showed higher desirable response rates on ptosis, ophthalmoplegia and general weakness at 3-6 months after treatment than other groups, and 42. 1％( 16/38 ) of them at 3 months achieved the desired state of ptosis, higher than the symptomatic group (7/48,14. 6％, ×2 = 8. 200, P = 0. 004 ). ② Ascending ideal rates had been presented in both combination and thymectomy groups since 1 year after treatments, while a little bit higher rate was presented in the former. At the end of observation, 21.7％ ( 13/60 )of patients in thymectomy group achieved complete stable remission.By paired longitudinal comparisons,thymectomy group showed higher ideal rates on ptosis (22/40,55.0％ ), ophthalmoplegia ( 16/27,59. 3％ ) and general weakness (20/40,50. 0％ ) at 2 years than that at 3 months( 11/59,18.6％ ;11/44,25.0％ ;9/60,15.0％ ;P =0. 002, 0. 031,0.000). ③For those patients by symptomatic treatment, the average age of onset was (51.9 ± 18.0) years, higher than that by other 3 therapies (F = 10. 563 ,P =0. 000). ④OMG patients with ophthalmoplegia more likely select corticosteroid or combined therapy. Ophthalmoplegia in combination group was higher than that in symptomatic and surgery groups( ×2 = 12. 939,14. 380, P =0. 000 in both). Ophthalmoplegia in corticosteroid group was higher than that in surgery group ( ×2 = 8. 017, P = 0. 005 ).Conclusions Corticosteroid appears to early overcome ptosis, ocular motor dysfunction and general weakness for patient with OMG in early-to-middle adulthood.Thymectomy andsurgery-corticosteroid combinationtherapies bothshowlong-term effectonthem.

Objective To explore thc clinical manifestation of secondary generalized myasthenia gravis(GMG) and analyze the factors affecting the progression from ocular myasthenia gravis(OMG) to GMG.Methods This research constitutes a single-center,retrospectively-collected prospective cohort study.We comprehensively reviewed our self-managed myasthenia gravis (MG) database drawn from personal clinical experience from January 2000 to Junc 2013.Patients underwent series of examination including repetitive nerve stimulation (RNS) tests,measurement of serum acetylcholine receptors antibody and serum muscle-specific tyrosine kinase antibodies,thymus computer tomography scan etc.Patients were treated with pyridostigmine bromide,corticosteroid therapy and (or) thymectomy based on a nonrandomization pattern and they were documented for their respective symptoms of OMG and GMG and date of GMG conversion.Logistic regression analysis was adopted to determine the influencing factors correlated with the development of GMG during the follow-up.Results Totally 770 patients initially diagnosed with OMG were included,among whom 573 (74％) patients remained with OMG (R-OMG group) and 197(26％) patients developed into GMG (GMG group) during the follow-up.(1) In comparison with their R-OMG counterparts,patients with secondary GMG were older at onset; Displayed more frequent RNS abnormality of facial nerve,accessory nerve and ulnar nerve ; Showed higher incidence of thymoma and were less treated by early corticosteroids.(2) For GMG group,81％ (160/197) of them displayed bulbar MG; 67％ (132/197) of GMG conversion occurred within 2 years,and 84％ (166/197) within 5 years.In comparison with the patients with onset of≤ 14-year-old,both of patients with15-49-year-old and≥ 50-year-old displayed higher conversion rate and shorter conversion duration (median:10 years versus 1 year and 6.5 months).(3) RNS abnormality of accessory nerve(OR =6.650,95％ CI 3.547-12.471 ; P ＜ 0.05) and thymoma(OR =7.924,95％ CI 2.554-24.585 ; P ＜ 0.05) were prognostic factors for the development of GMG,while early corticosteroid(OR =0.232,95％ CI 0.119-0.452 ; P ＜ 0.05) predicted the reduction of the risk of generalization.Conclusions Multiple factors including abnormal RNS of proximal limb muscles,thymoma,early corticosteroids therapy and possibly even onset age of over 15-year-old may involve the generalization in patients with OMG at onset.

[ ABSTRACT] The splicing factors were characterized for their crucial roles in pre-mRNA splicing of eukaryons. SRSF2 is a member of the SR protein family which is one of the most common splicing factors, and it is believed to be a key element in pre-mRNA splicing, mRNA transcription, regulation of the DNA stability and cell proliferation.SRSF2 gene mutation is detected frequently in myeloid malignancies ( like MDS and CMML) and may be associated with the phenotype and prognosis of these malignancies.The paper makes a review for the latest research progression on SRSF2 gene mutation and its relationship with myeloid malignancies.

Objective To explore the changes in expression of Klotho, an aging-suppression protein, and neutrophil gelatinase associated lipocalin ( NGAL) and their relationship with rat mesangial cells ( RMCs) cultured with high glucose in vitro, and to explore the role played by Toll-like receptor-4 (TLR4) / nuclear factor-kB(NF-kB) p65 pathways in this process. Methods Three NGAL-siRNA sequences were designed and synthesized. The effective sequence in subsequent experiments was chosen. RMCs were preincubated with pyrrolidinedithiocarbamate (PDTC)or exogenously added Klotho prior to high glucose treatment. Realtime PCR was used to analyze Klotho, TLR4, NGAL mRNA expressions. Western blot was used to observe Klotho, TLR4,NF-kB p65, NGAL,fibronectin (FN), and connective tissue growth factor ( CTGF) protein expression. ELISA assay was used to detect monocyte chemoattractant protein-1 ( MCP-1) and CXCL5 secretions. Results High glucose suppressed Klotho expression significantly(P<0. 05) and activated TLR4 / NF-kB p65 pathway. Meanwhile,the levels of NGAL,FN,CTGF, MCP-1, and CXCL5 were highly expressed ( P < 0. 01). NGAL gene silencing obviously down-regulated the increased expressions of FN, CTGF, MCP-1, and CXCL5 ( P < 0. 01). After PDTC treatment the overexpression of NGAL protein was markedly lowered(P<0. 01). In addition, Klotho treatment significantly inhibited the activity of TLR4 /NF-kB p65 pathways and down-regulated the expressions of NGAL, FN, CTGF, MCP-1 and CXCL5 stimulated by high glucose(P<0. 01). Conclusion Klotho inhibits the activity of TLR4 / NF-kB p65 pathways and thus inhibits NGAL expression in RMCs cultured with high glucose in vitro. And then it suppresses the expressions of FN, CTGF, MCP-1, and CXCL5. This provides a new basis to illustrate the protection mechanism of the anti-aging protein Klotho in diabetic nephropathy, and may provide new ideas and therapeutic targets for prevention and treatment.

Objective? To explore the role of optimizing health education in regulating the use of antimicrobial agents for outpatient patients in infection department. Methods? Using convenience sampling method, 400 patients with moderate infection who needed intravenous antibiotics in the infectious Outpatient Department of Zhongnan Hospital of Wuhan University from January to December 2017 were selected and divided into experimental group and control group with 200 cases in each group according to random number table method. The patients in the control group were given routine nursing care, while the patients in the experimental group were given optimized health education intervention on the basis of routine nursing care. After the treatment, the knowledge, compliance and long-term efficacy of antimicrobial drugs were compared between the two groups. Results? Before intervention, there was no significant difference in the knowledge of antimicrobial drugs between the two groups (P＞0.05); after intervention, the knowledge level of antimicrobial drugs in the two groups was improved, and the scores of the dosage of antimicrobial drugs, the principle of drug use, adverse reactions, emergency measures and the total score of knowledge in the experimental group were higher than those in the control group with statistically significant differences (P＜0.01). Compliance rates of drug use on time, reasonable diet, timely re-visit, smoking cessation and alcohol cessation in the experimental group were 95.0%, 93.5%, 97.0% and 96.0%, respectively, which were higher than those in the control group (80.0%, 85.0%, 78.0% and 84.0%, respectively) with statistically significant differences (P＜ 0.01). After 6 months of intervention, the recurrence rate in the observation group was 4%, lower than the control group with 10%, and the difference was statistically significant (P＜ 0.05). Conclusions? Optimized health education can effectively improve the knowledge level of antibiotics in outpatients of infectious department, improve the compliance of patients and reduce the recurrence rate, which is worthy of clinical application.

Objective:To investigate the effect of long non-coding RNA (lncRNA) H19 regulating miRNA-675 (miR-675) and phosphatase and tensin homologue-deleted chromosome ten gene (PTEN) on the cell proliferation of glioma.Methods:Glioma cell lines U87-MG and U251 were chosen. The siRNA online design tool wad used to design small interfering RNA (siRNA) targeting H19. U87-MG and U251 cell lines with the stable knockdown of H19 were constructed (the stable knockdown of H19 group), and the cells randomly transfected with siRNA plasmid were taken as the control group, and normal cultured cells were treated as the blank group. Additionally, miR-675 and control microRNA were transfected into U87-MG and U251 with the stable knockdown of H19 (the overexpressing miR-675 group and the corresponding control group). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of miR-675 and H19 in each group; the methyl thiazolyl tetrazolium (MTT) assay was used to detect the cell proliferation ability; the dual luciferase reporter gene assay was used to verify the targeting relationship between miR-675 and PTEN; Western blot was used to detect the relative expression level of PTEN protein.Results:The MTT assay results showed that the proliferation ability of U87-MG and U251 cells in the stable knockdown of H19 group was lower than that of the corresponding control group; and the differences in cell proliferation ability of all the groups after 48 h of culture were statistically significant (all P < 0.05). qRT-PCR detection results showed that the relative expression level of miR-675 in U251 cells in the stable knockdown of H19 group and the corresponding control group was 0.329±0.009 and 1.043±0.087, respectively, and the difference was statistically significant ( t = 14.15, P < 0.001); the relative expression level of miR-675 in U87-MG cells in the stable knockdown of H19 group and the corresponding control group was 0.299±0.009 and 1.027±0.106, respectively, and the difference was statistically significant ( t = 11.85, P < 0.001); the relative expression level of miR-675 in U87-MG and U251 cells in the stable knockdown of H19 group was lower than that of the corresponding control group. The dual luciferase reporter gene assay verified that miR-675 could bind to the 3'-UTR of PTEN. Western blot detection results showed that the relative expression level of PTEN protein in U87-MG and U251 cells in the stable knockdown of H19 group was higher than that of the corresponding control group and the blank group; in the U87-MG and U251 cells in the stable knockdown of H19 group, the relative expression level of PTEN in the overexpressing miR-675 group was lower than that of the corresponding blank group and the control group. In the U87-MG and U251 cells in the stable knockdown of H19 group, the cell proliferation ability of the overexpressing miR-675 group was higher than that of the corresponding blank group and the control group; the differences in cell proliferation ability of all the groups after 48 h of culture were statistically significant (all P < 0.05). Conclusions:lncRNA H19 may regulate the cell proliferation of glioma cells through the miR-675-PTEN signaling pathway.

Objective:To elucidate the change of whole genome expression profile for the effect of melatonin on radiation-induced intestinal injury in mice.Methods:C57BL/6J male mice were administrated with melatonin at 10 mg/kg body weight by intraperitoneal injection once a day for five consecutive days before abdominal irradiation with 14 Gy of γ-rays. Small intestines were harvested 3 d after radiation. GO annotation and KEGG pathway of the differential genes involved in small intestine were explored by DNA microarray analysis.Results:Compared with the control group, 584 differential genes were up-regulated and 538 differential genes were down-regulated for administration group pre-irradiation. The overlapping differential genes were selected from the irradiated mice and the administrated mice pre-irradiation. There were 324 up-regulated genes and 246 down-regulated genes unique to the administrated mice pre-irradiation. GO annotation analysis of the differential genes indicated that the top 15 significantly enriched biological processes for the administrated mice pre-irradiation mainly included autophagosome assembly (GO: 0000045), autophagosome organization (GO: 1905037) and regulation of acute inflammatory response (GO: 0002673). The genes ATG12, ATG16L2 and AMBRA1 were involved in autophagosome assembly and autophagosome organization. The genes C3, CPN1, CD55, CFP, CNR1, C1QA, C2 and CREB3L3 were involved in the regulation of acute inflammation response. KEGG pathway analysis of the differential genes involved indicated that the top 15 significantly enriched pathways for the administrated mice pre-irradiation mainly included O-glycan biosynthesis (hsa00512), glycosphingolipid biosynthesis (hsa00603), ECM-receptor interaction (hsa04512) and biosynthesis of unsaturated fatty acids (hsa01040). qRT-PCR verification showed that the expressions of ATG12 and ATG16L2 genes involved in autophagy for the administrated mice pre-irradiation increased significantly compared with the irradiated mice ( t=2.40, 4.35, P<0.05). Conclusions:The differential genes related with the biological process of autophagy, acute inflammatory response and the pathway of unsaturated fatty acid biosynthesis might be involved in the effect of melatonin on radiation-induced intestinal injury.