OBJECTIVETo verify whether abnormal expression of calcium/calmodulin dependent serine protein kinase (CASK) and inhibitors of differentiation 1 (ID1) exist in Fb of keloid, and to observe the effect of artesunate on two genes.

METHODSFifteen samples of keloid and 12 samples of normal skin tissue (discarded) excised from patients admitted to our hospital were collected. Tissue particle adherent method was used in the primary culture of Fb, and cells from the third to the eighth passage were used for test. Expressions of CASK and ID1 in Fb harvested from both sources were observed with immunofluorescence staining. Fb of keloid were stimulated with artesunate in various concentration for different time, and the median inhibitory concentration (IC50) was determined with the MTT colorimetric assay, which served as the intervention concentration of artesunate. Fb of normal skin were set as normal control group (NC, treated with medium solution). Fb of keloid were divided into scar control group (SC, treated with medium solution) and scar administration group (SA, treated with artesunate in IC50). The cycle and apoptosis of Fb were detected with flow cytometric assay, and the nucleic acid and protein expressions of CASK and ID1 of Fb in each group were determined with RT-PCR and Western blotting. Data were processed with one-way analysis of variance and LSD-t test.

RESULTSExpressions of CASK and ID1 were detected in two kinds of Fb. The concentration of 75 mg/L was selected as the intervention concentration of artesunate. (1) There were statistically significant differences among the three groups in the percentages of cells in G0/G1 phase and G2/M phase (with F values respectively 118.064 and 163.840, P values all below 0.01). The percentage of cells in G0/G1 phase of group SA was (91.4 ± 1.4)%, which was significantly higher than that of group SC and group NC [respectively (80.7 ± 0.3)% and (82.4 ± 0.6)%, with t values respectively 12.740 and 9.872, P values all below 0.05]. The percentage of cells in G2/M phase of group SA was (6.9 ± 0.3)%, which was significantly lower than that of group SC and group NC [respectively (13.7 ± 0.3)% and (12.7 ± 0.8)%, with t values respectively 43.702 and 12.276, P values all below 0.05]. (2) There were statistically significant differences among the three groups in the early and late apoptotic rates (with F values respectively 61.879 and 4710.862, P values all below 0.01). The early and late apoptotic rates of group SA were respectively (7.1 ± 1.0)% and (14.9 ± 0.3)%, which were significantly higher than those of group SC and group NC [with early apoptotic rate respectively (2.6 ± 0.4)% and (2.7 ± 0.3)%, t values respectively 7.974 and 7.767, P values all below 0.05; with late apoptotic rate respectively (2.3 ± 0.3)% and (2.5 ± 0.4)%, t values respectively 72.882 and 69.792, P values all below 0.05]. (3) The mRNA expression of CASK in group SC was 0.658 ± 0.024, and it was lower than that of group NC (1.076 ± 0.008, t = 28.997, P < 0.01) and group SA (0.855 ± 0.008, t = 13.549, P < 0.01). The protein expression of CASK in group SC was 0.067 ± 0.007, and it was lower than that of group NC (0.179 ± 0.015, t = 12.042, P < 0.01) and group SA (0.132 ± 0.010, t = 9.498, P < 0.01). (4) The mRNA expression of ID1 in group SC was 0.416 ± 0.006, which was higher than that of group NC (0.317 ± 0.020, t = 8.299, P < 0.01) and group SA (0.217 ± 0.009, t = 32.417, P < 0.01). The protein expression of ID1 in group SC was 0.789 ± 0.034, and it was higher than that of group NC (0.366 ± 0.029, t = 16.341, P < 0.01) and group SA (0.114 ± 0.006, t = 33.978, P < 0.01).

CONCLUSIONSIt is speculated that CASK and ID1 participate in the proliferation of Fb in keloid. The mechanism of artesunate in inhibiting the proliferation of Fb in keloid may be related to the up-regulation of CASK and down-regulation of ID1.

Adolescent ; Adult ; Apoptosis ; drug effects ; Artemisinins ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Female ; Fibroblasts ; metabolism ; Gene Expression Regulation ; Guanylate Kinases ; genetics ; metabolism ; Humans ; Inhibitor of Differentiation Protein 1 ; genetics ; metabolism ; Keloid ; metabolism ; pathology ; Male ; Middle Aged ; Young Adult

BACKGROUNDIn response to the injury of the central nervous system (CNS), the astrocytes upregulate the expression of glial fibrillary acidic protein (GFAP), which largely contributes to the reactive gliosis after brain injury. The regulatory mechanism of this process is still not clear. In this study, we aimed to compare the ephrin-B2 deficient mice with the wild type ones with regard to gliosis after traumatic brain injury.

METHODSWe generated ephrin-B2 knockout mice specifically in CNS astrocytes. Twelve mice from this gene-knockout strain were randomly selected along with twelve mice from the wild type littermates. In both groups, a modified controlled cortical impact injury model was applied to create a closed traumatic brain injury. Twenty-eight days after the injury, Nissl staining and GFAP immunofluorescence staining were used to compare the brain atrophy and GFAP immunoreactivity between the two groups. All the data were analyzed by t-test for between-group comparison.

RESULTSWe successfully set up the conditional ephrin-B2 knockout mice strain, which was confirmed by genotyping and ephrin-B2/GFAP double staining. These mice developed normally without apparent abnormality in general appearance. Twenty-eight days following brain injury, histopathology revealed by immunohistochemistry showed different degrees of cerebral injuries in both groups. Compared with wild-type group, the ephrin-B2 knockout group exhibited less brain atrophy ratio for the injured hemispheres (P = 0.005) and hippocampus (P = 0.027). Also the wild-type group demonstrated greater GFAP immunoreactivity increment within hippocampal regions (P = 0.008).

CONCLUSIONSThe establishment of conditional ephrin-B2 knockout mice provides us with a new way to explore the role of ephrin-B2 in astrocytes. Our findings revealed less atrophy and GFAP immunoreactivity in the knockout mice strain after traumatic brain injury, which implied ephrin-B2 could be one of the promoters to upregulate gliosis following brain injury.

Animals ; Atrophy ; Brain ; pathology ; Brain Injuries ; complications ; pathology ; Ephrin-B2 ; deficiency ; physiology ; Glial Fibrillary Acidic Protein ; Gliosis ; etiology ; Immunohistochemistry ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Nerve Tissue Proteins ; analysis

The study was aimed to investigate the potential immunotherapeutical values of umbilical cord tissue-derived mesenchymal stem cells (UC-MSC) on patients with chronic idiopathic thrombocytopenic purpura (ITP). UC-MSC was cocultured in vitro with splenocytes isolated from ITP patients who experienced splenectomy. The level of IgG antiplatelet antibody (PAIgG) was determined by a competitive micro-enzyme-linked immunosorbent assay (ELISA) method. The proliferation of platelet-reactive CD4+ T lymphocytes was also measured in the presence of UC-MSCs. The results showed that UC-MSCs could stimulate the spontaneous secretion of PAIgG in supernatants; In the platelet-inducing condition, UC-MSC inhibited the production of PAIgG at a low ratio of 1 UC-MSC to 100 splenocytes, but promoted at a high proportion of 1 UC-MSC to 10 splenocytes. Moreover, UC-MSC exerted a suppressive effect on proliferation of platelet-reactive T helper cells in a dose-dependent manner. It is concluded that the UC-MSCs can regulate secretion of antiplatelet antibodies in vitro. Its concrete regulation mechanism and potential immunotherapeutical value are need to further study.
Antibodies ; metabolism ; Blood Platelets ; immunology ; CD4-Positive T-Lymphocytes ; cytology ; Cell Proliferation ; Humans ; Infant, Newborn ; Lymphocyte Activation ; Mesenchymal Stromal Cells ; physiology ; Purpura, Thrombocytopenic, Idiopathic ; metabolism ; Spleen ; cytology ; Umbilical Cord ; physiology

OBJECTIVESTo investigate if hyperinsulinemia or insulin resistance could predict the elevation of blood pressure in non-diabetic adults.

METHODSOne hundred and seventy non-diabetic adults (NGT 107, IGT 63) were included based on the screen by OGTT in 1986. Height, weight, blood pressure were measured. Plasma glucose and insulin concentration at 0.60 and 120 min during OGTT were determined at baseline. All the subjects were followed for six years with blood pressure and plasma glucose examined at the end of the study. Subjects worsening to diabetes were excluded. Insulin area under-curve (INSAUC) and insulin sensitivity index [IAI = (1/FINS x FPG)] were calculated. Stepwise regression analysis was performed to evaluate the effects of INSAUC and insulin sensitivity to the elevation of blood pressure.

RESULTSBoth SBP and DBP levels at the end of the study were increased with increased INSAUC baseline. The SBP were (119.5 +/- 2.3), (122.1 +/- 2.5), (129.4 +/- 2.4) and (128.3 +/- 2.6) mmHg, and the DBP were (78.6 +/- 1.6), (79.7 +/- 1.7), (85.2 +/- 1.4) and (84.0 +/- 1.0) mmHg from the lowest to the highest quartiles of INSAUC respectively. Pearson correlation analysis showed Age, SBP, DBP, BMI, FINS, INS1h, INSAUC at baseline were positively correlated to blood pressure levels at the end of the study. After the adjustment of Age, sex, BMI, smoking, PG2 h and blood pressure at baseline, the INSAUC was significantly correlated to blood pressure six years later, while the insulin sensitivity index was not.

CONCLUSIONThe compensated hyperinsulinemia based on selective insulin resistance rather than insulin resistance to glucose per se could predict the elevation of blood pressure in nondiabetic adults.

Adult ; Aged ; Blood Pressure ; physiology ; Diabetes Complications ; Diabetes Mellitus ; prevention & control ; Female ; Follow-Up Studies ; Glucose Intolerance ; blood ; Glucose Tolerance Test ; Humans ; Hyperinsulinism ; complications ; Hypertension ; etiology ; prevention & control ; Insulin Resistance ; Male ; Mass Screening ; Middle Aged

OBJECTIVETo investigate the human immunodeficiency virus (HIV) and syphilis incidence rates as well as the retention rate in a cohort with 6-month follow-up study among female sex workers (FSWs).

METHODSFrom December, 2004, to January, 2005, a community-based baseline survey was conducted to recruit 343 FSWs for a prospective cohort study in Xichang county of Sichuan province, China. Follow-up visits were conducted at 6 months to analyze risk factors associated with cohort retention for subjects' baseline sociodemographic and sexual behavioral characteristics. Blood specimens were also collected to test antibodies against HIV and syphilis.

RESULTSDuring the 6-month follow-up period, HIV and syphilis incidence appeared to be 1.00 per 100 person-years and 6.23 per 100 person-years, respectively. The rate of retention in the cohort was 53.6% (184/343). Results from multivariate logistic regression model showed that factors were significantly associated with cohort retention including people with minority ethnic background (OR = 0.36; 95% CI: 0.18-0.74), people having participated in AIDS prevention program (OR = 1.83; 95% CI: 1.17-2.86) or being clients in the last 6 months > or = 50 (OR = 1.75; 95% CI: 1.11-2.77) and having changed living/working place (OR = 0.56; 95% CI: 0.33-0.94).

CONCLUSIONThe results of this study showed that the syphilis incidence and unprotected sex behavior were high among local FSWs. People belonged to Han nationality, having participated in AIDS prevention program and having a steady living/working place were associated with cohort retention at 6-month follow-up study among FSWs, respectively.

China ; epidemiology ; Female ; Follow-Up Studies ; HIV Infections ; epidemiology ; Humans ; Incidence ; Risk Factors ; Sex Work ; Sexual Behavior ; Substance-Related Disorders ; Syphilis ; epidemiology

Gynandromorphic ticks are extremely rare, and often attract parasitologists' attention. During our examination of tick specimens, an engorged gynandromorph of Hyalomma asiaticum was noticed. This is the first record of gynandromorphic ticks from China. In this study, several important morphological structures of normal and gynandromorphic H. asiaticum were analyzed. Comparing to the normal H. asiaticum, the gynandromorphic specimen was a typical bipartite protogynander. Its right side showed normal female characteristics, whereas the left side had normal male traits. Different from other gynandromorphic ticks containing 1 anus, this tick reported here had 2 complete anuses, and the anus of the male part had a single adanal plate.
Animals ; Chimera/*anatomy & histology/genetics ; China ; Female ; Ixodidae/*anatomy & histology/genetics ; Male ; Sheep ; Sheep Diseases/*parasitology ; Tick Infestations/parasitology/*veterinary

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cell Culture Techniques ; methods ; Child ; Female ; Humans ; Induced Pluripotent Stem Cells ; cytology ; metabolism ; Male ; Middle Aged

Animals ; Cellular Reprogramming ; Dentate Gyrus ; cytology ; Fibroblasts ; cytology ; Mice ; Neural Stem Cells ; cytology ; transplantation ; Swine

Objective:To systematically investigate the chemical constituents of 90% ethanol extract of Corydalis impatiens，and evaluate their inhibitory effect on cell proliferation in vitro. Method:The chemical constituents were isolated and purified by silica gel，Sephadex LH-20，ODS column chromatography and semi-preparative high performance liquid chromatography （HPLC）.the structures were identified by spectroscopy methods such as NMR and MS，as well as analysis of physicochemical properties and/or comparison with literature data，and the inhibitory activities of 13 compounds on HepG2 and SMMC-7721 cells were measured by methyl thiazolyl tetrazolium （MTT） method. Result:Thirteen known compounds were isolated and identified from 90% ethanol extract of Corydalis impatiens as ethyl-5-hydroxy-2-pyridinecarboxylate（1），coryhumolide（2），3，4-cis-3，4-dihydroxy-β-ionone（3），megastigmane（4），9-hydroxy-4，7-megastigmadien-3-one（5），blumenol A（6），indole-3-carboxy acid（7），1-methyl-[1，2，4]triazolo[4，3-b][1，2，4]triazin-7-one （8），adenine（9），nicotinamide（10），2-hydroxymethyl-5-pyridinol（11），adenosine（12）， and β-daucosterol（13）. Cytotoxicities assay showed that the IC₅₀ value of compound 3 for the hepatic cell line HepG2 was 24.7 μmol·L^-1 （positive control drug cisplatin：4.8 μmol·L^-1），and IC₅₀ value of compound 4 for the hepatic cell line SMMC-7721 was 13.8 μmol·L^-1（positive control drug cisplatin：5.4 μmol·L^-1）. Conclusion:Compound 1 was a new natural compound，compounds 3-8 were obtained from genus Corydalis for the first time，and compounds 2，9-12 were isolated from this plant for the first time. Compound 3 exhibited weak inhibitory effect on hepatic cell line HepG2，and compound 4 exhibited moderate inhibitory effect on hepatic cell line SMMC-7721.The other compounds rest of ones exhibited no obvious inhibitory effect on hepatic cell line HepG2 or SMMC-7721.