Drugs, Chinese Herbal ; therapeutic use ; Humans

Objective To investigate the role of stromal cell derived factor-1α (SDF-1α)-laminin (LN) crosstalk in migration and differentiation of neural stem cells (NSCs).Methods Original generation of NSCs collected from 14-day pregnant fetal rats were separated and cultivated, and the phenotype characteristics were identified with immunofluorescence. The third generation of NSCs were collected, and they were divided into four groups: poly-L-lysine (PLL) group, PLL+SDF-1α group, LN group and LN+SDF-1α group. The NSCs in PLL and LN groups were inoculated into plates coated with 0.1 mg/mL PLL or 1 mg/mL LN, and the NSCs in PLL+SDF-1α and LN+SDF-1αgroups were inoculated into plates coated with PLL or LN and 1 mg/mL SDF-1α containing medium. The effect of SDF-1α-LN crosstalk on NSCs migrated numbers was determined by using Transwell cell migration test, and the differentiation of NSCs was determined by immunofluorescence staining.Results Primary NSCs were successfully isolated and cultivated, neurospheres formed at 3-5 days with typical NSCs morphology positively expressing Nestin which was the specific antigen of NSCs. Compared with PLL group, the number of NSCs migration in PLL+SDF-1α group showed no significant change (cells: 3.00±0.99 vs. 2.3±0.67,P > 0.05). Compared with LN group, the number of NSCs migration in LN+SDF-1α group was significantly increased (cells: 85.33±9.61 vs. 31.67±5.86,P < 0.05). Immunofluorescence staining showed that the differentiation rates of PLL and PLL+SDF-1α groups were almost zero, and some early differentiation neurons were detected in LN group with the differentiation rates of (12.50±2.56)%. The differentiation of early neuronal cells was significantly increased after SDF-1α-LN crosstalk, the neuronal differentiation rate was (21.40±3.41)%, and it was significantly higher than that of LN group (P < 0.05).Conclusion SDF-1α crosstalk with LN in extracellular matrix can significantly and synergistically enhance the migration and differentiation of NSCs in vitro.

Reflux esophagitis is a common type of gastroesophageal reflux disease.Its pathogenesis is not completely clear,but the body immunity,oxidative stress and chemical damage have gradually become the research focus.According to the latest research, the expressions of related proteins in reflux esophagitis provide a foundation for its pathogenesis. And some kinds of proteins can be used as monitoring index in the development process from reflux esophagitis to esophageal adenocarcinoma. This article is to review the expression and significance of proteins in reflux esophagitis in recent years.

Objective To investigate the genetic polymorphism of mec Ⅰ in the clinical isolates of methicillin-resistant Staphylococcus anreus(MRSA).Methods 40 isolates(MRSA)carrying mecA gene were selected randomly from the clinical isolates of Staphylococcus anreus from Jan,2005 to Aug,2006 in our hospital.The mec Ⅰ gene was detected by PCR followed with sequencing.Staphylococcal cassette chromosome mec(SCCmec)in MRSA were detected by multiplex-PCR.Agar dilution method was used for determining the MICs of oxacillin against MRSA.Results 35 of 40(87.5%)MRSA carried mec Ⅰ gene.All isolates carrying mec Ⅰ gene have mecI 202C→T substitution,which resulted in Gln at 68 aminophenol position replaced by stop condon.32 isolates carried single point mutation.3 isolates carried double-point mutation,including additonal A at 3 positon,A→C at 41 position and C→T at 142 position beside C→T at 202 position,respectively.Among 35 isolates carrying mec Ⅰ gene,there were 27 isolates of SCCmec Ⅲ, 7 isolates of SCCmec Ⅲ A and 1 isolate of SCCmec Ⅱ.Among 5 isolates with deletion of mec Ⅰ gene,there were 3 isolates of SCCmecⅣ,1 isolate of SCCmec Ⅰ and 1 isolate of non-known SCCmec tpye.The MICs of oxacillin were 256-512 ?g/ml,≥512 ?g/ml and 8-256 ?g/ml in 31 isolates with single point mutation at 202 position in mec Ⅰ gene,3 isolates with double-point mutation in mecI gene and 5 isolates with deletion of mec Ⅰ gene,respectively.1 isolate with single point mutation in mec Ⅰ gene had contrary result(MIC

In order to establish K562 line with stable expressions of hermap and hermap-siRNA, amplified hermap and hermap-siRNA were cloned into pEGFP-c1 and pRNAT to acquire hermap-pEGFP-c1 and hermap-siRNA-pRNAT, respectively. These two plasmids were electrotransferred into K562 cells, then were followed by culturing with G418. The result showed that the transfer rate of hermap-pEGFP-c1-K562 and hermap-siRNA-pRNAT-K562 plasmids were 10.0% and 9.3%, respectively. After selective culture by G418, these two cell lines were still able to express GFP. It is concluded that the eukaryotic expression plasmids containing hermap and hermap-siRNA have been successfully constructed, and the cell lines of hermap-K562 and hermap-siRNA-K562 are established, definitely contributing to further functional investigation on HERMAP and its interaction with other proteins.
Erythrocytes ; chemistry ; Gene Silencing ; Humans ; K562 Cells ; Membrane Proteins ; genetics ; Plasmids ; RNA, Small Interfering ; Transfection

Objective To understand the clinical features and prognosis of children with severe viral encephalitis (SVE), evaluate the related factors affecting prognosis. Methods Clinical data of 102 children with SVE in pediatric neurological ward and pediatric intensive care unit in Hunan Children's Hospital between January 2014 and January 2016 were analyzed retrospectively. According to prognosis, children were divided into good prognosis group(n =24, children's Glasgow outcome scale[CGOS]: 4 — 5) and poor prognosis group(n = 78, CGOS: 1 - 3), clinical data of two groups of children were compared, risk factors affecting the prognosis of SVE children were analyzed. Results In good prognosis group, 15 cases were cured and 9 had mild sequelae; in poor prognosis group, 14 cases died, 25 had severe sequelae, and 39 had moderate sequelae. The duration of fever and length of hospital stay in good prognosis group were both shorter than poor prognosis group, difference was statistically significant (both P く0.05). Multivariate unconditioned logistic regression analysis showed that adverse factors for prognosis of SVE were as follows: convulsive status, respiratory failure, longer fever period(＞5 days), severely abnormal electroen-cephalogram(EEG), head magnetic resonance imaging (MRI) lesions involving more than two sites or lesions involving the infratentorial, and stress hyperglycemia, odds ratio(OR) were 13.468, 4.580, 2.378, 10.196, 3.012, and 6.316 respectively. Conclusion SVE is a serious threat to quality of children's life, convulsive status, respiratory failure, longer fever period, severely abnormal EEG, head MRI lesions involving more than two sites or lesions involving the infratentorial, and stress hyperglycemia are risk factors for prognosis of SVE in children.

Amorphous solid dispersion (ASD) is one of the most effective formulation approaches to enhance the water solubility and oral bioavailability of poorly water-soluble drugs. However, maintenance of physical stability of amorphous drug is one of the main challenges in the development of ASD. Crystallization is a process of nucleation and crystal growth. The nucleation is the key factor that influences the physical stability of the ASD. However, a theoretical framework to describe the way to inhibit the nucleation of amorphous drug is not yet available. We reviewed the methods and theories of nucleation for amorphous drug. Meanwhile, we also summarized the research progress on the mechanism of additives influence on nucleation and environmental factors on nucleation. This review aims to enhance the better understanding mechanism of nucleation of amorphous drug and controlling over the crystal nucleation during the ASD formulation development.

OBJECTIVETo study the effects of hermap gene on kinases in erythroid signal transduction pathway and investigate the mechanism of hermap on erythroid differentiation.

METHODSThe K562 cells expressing hermap and hermap-siRNA respectively were established for up- and down-regulating the expression of hermap gene. These K562 cells were then induced by Ara-C to erythroid differentiation and analyzed at 0, 24, 48, 72 and 96 h, respectively, for cell morphology and biphenylamine staining positive cells, determination of CD235a, CD36, kinases p-STAT5, p-Akt, p-MAPK and p-c-JUN by FCM; and quantification of hermap gene and γ (Aγ,Gγ) globin gene by FQ-PCR.

RESULTSWith up-regulating hermap gene and inducing by Ara-C, K562 cells were changing to low ratio of nucleus to cytoplasm, cytoplasm colour from basophilic to pinkish or amethyst tinge, increase of number of biphenylamine positive cells and expression of CD235a, CD36, γ (Aγ,Gγ) globin gene, hermap gene and p-STAT5 from 0 to 96 h. At 0, 24, 48, 72 and 96 h of culture, the positive rates of p-STAT5 cells were detected of 0.46%, 4.54%, 20.01%, 23.65% and 33.08%, respectively. This results demonstrated that there was a positive correlation between expression of p-STAT5 and hermap gene expression (P < 0.05).

CONCLUSIONhermap gene can stimulate erythroid differentiation of Ara-C induced K562 cells mainly through JAK/STAT5 signal transduction pathway.

Cell Differentiation ; Erythrocyte Membrane ; Erythrocytes ; cytology ; Erythropoiesis ; Gene Expression ; Humans ; K562 Cells ; Receptors, Erythropoietin ; genetics ; STAT5 Transcription Factor ; metabolism ; Signal Transduction

OBJECTIVETo investigate the necessity of detecting on the expressive intensity and pattern of HBsAg and HBcAg in the livers of chronic hepatitis B.

METHODSHBsAg and HBcAg were detected in paraffin-embedded liver tissue by EnVision immunohistochemistry. Serum hepatitis B virus DNA (HBV DNA) was tested by real-time quantitative polymerase chain reaction. The degrees of hepatic inflammatory activity (grade) and fibrosis (stage) of liver biopsies were determined according to the standard of the Chinese program of prevention and treatment of viral hepatitis.

RESULTSThe expression of HBsAg was not correlated with the grade, the stage and the levels of serum HBV DNA (P > 0.05). Liver HBcAg expressive intensity was not correlated with the grade (r=0.02, P > 0.05), while negatively correlated with the stage (r=0.28, P < 0.01) and positively correlated with the serum HBV DNA levels (r=0.53, P < 0.01). Liver HBcAg expressive pattern was negatively correlated with the grade (r=-0.27, P < 0.01). The grade in cytoplasmic pattern group was higher than in nuclear pattern group and in mixed pattern group (P < 0.01), and that in mixed pattern group was higher in nuclear pattern group (P < 0.01). Liver HBcAg expressive pattern was negatively correlated with the stage (r=-0.23, P < 0.01). The stage in cytoplasmic pattern group was higher than in nuclear pattern group and in mixed pattern group (P < 0.05). Liver HBcAg expressive pattern was positively correlated with the levels of serum HBV DNA (r=0.22, P < 0.01).

CONCLUSIONDistinguishing the expressive intensity and pattern of HBsAg and HBcAg in the liver of chronic hepatitis B may not help understand the degree of hepatic lesion. The detection of HBcAg in liver tissue of CHB may be beneficial for the antiviral therapy.

Adult ; DNA, Viral ; blood ; genetics ; Female ; Hepatitis B Antigens ; biosynthesis ; Hepatitis B Core Antigens ; biosynthesis ; Hepatitis B Surface Antigens ; biosynthesis ; Hepatitis B virus ; genetics ; immunology ; physiology ; Hepatitis B, Chronic ; pathology ; virology ; Host-Pathogen Interactions ; Humans ; Immunohistochemistry ; Liver ; pathology ; virology ; Male ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction ; Virus Replication ; Young Adult

Aim To determine the effect of exosomes from lipopolysaccharide-treated human bone marrow mesenchymal stem cells on proportion of Ly6Chigh and Ly6Clow monocytes/macrophages in inflammatory micro- environment. Methods BMSCs were obtained by gra-dient centrifugation, identified and then treated with li-popolysaccharide for 48 h. The exosomes were purified from conditional medium with or without LPS treatment and identified by CD63 protein using Western blot and transmission electron microscope. The diameters and concentration were detected by Nanoparticle Trafficking Analysis ( NTA ) . The monocytes/macrophages were sorted from bone marrow of the mice by magnetic beads. Cells were co-cultured with exosomes for 24 hours, and then treated with LPS for 48 hours. The proportion of Ly6C monocytes/macrophages was detec-ted by flow cytometry. Inflammatory cytokines in cell supernatant were investigated using ELISA. Results BMSCs surface markers CD44, CD90 were positively detected, but CD34, CD45 were not expressed. BM-SCs presented adipogenic differentiation ability. Exo-somes were positively expressing CD63 protein, and NTA showed that the diameters of exosomes were up to (82.4 ± 3.7 ) nm. BMSCs stimulated by LPS pro- duced more exosomes ( P < 0.01 ) . Exosomes from BMSCs with or without LPS treatment could increase the ratio of Ly6Chigh monocytes (P<0.01) and down-regulate the ratio of Ly6Chigh macrophages (P<0.05), and the effect of LPS treated-exosomes was more signif-icant than untreated-exosomes (P<0.05). Moreover, the concentration of IL-6 was also elevated under exo-somes treatment ( P <0.05 ) . Conclusions Human bone marrow mesenchymal stem cells-derived exosomes contribute to the regulation of Ly6Chigh monocytes/mac-rophages, indicating that they could be involved in the therapeutic treatment of inflammatory diseases.