Objective To explore the action mechanism and clinical significance of CD4+CD25+ regulatory T (Treg) cells in the development of atopic dermatitis (AD). Methods Peripheral blood mononuclear cells (PBMC) were obtained from 46 patients with AD and 20 normal human controls. Flow cytometry was performed to detect the number of CD4+CD25+ Treg cells, real-time fluorescence PCR assay to measure the Foxp3 mRNA level in PBMC, ELISA to determine the serum levels of IL-2, IL-4, IL-10 and IFN-γ. Results A statistical decrease was observed in the percentages of peripheral CD4+CD25+ Treg cells among CD3+ T cells and CD4+ T cells in AD patients compared with normal controls (t' = 3.775, 4.533, both P＜ 0.01 ), and in the percentage of peripheral CD4+CD25+ Treg cells among CD3+ T cells in patients with acute AD compared with those with chronic AD (t = 2.217, P ＜ 0.05), but no significant difference was noted between patients with acute AD and those with subacute AD or between those with subacute AD and those with chronic AD in the percentage of peripheral CD4+CD25+ Treg cells among CD3+ T cells (t = 1.558, 0.49, both P ＞ 0.05). The mRNA level of Foxp3 in PBMC from AD patients was statistically decreased compred with that from normal controls (z =-2.368, P ＜ 0.05 ). The count of CD4+CD25+ Treg cells was positively correlated with serum levels of IL-2 and IL-10 (r = 0.512, 0.494, both P ＜ 0.05), but had no significant correlation with serum levels of IL-4 and IFN-γ (r = -0.110, -0.237, both P ＞ 0.05). Conclusions In AD patients, there is a decrease in the count of CD4+CD25+ Treg cells and in the level of Foxp3 mRNA, which may suppress the proliferation of and secretion of Foxp3 mRNA by Th2 cells, lead to Th2 predominance, participate in the development of AD.

BACKGROUND: In vivo experiments have confirmed that fibroblast growth factor can effectively protect gentamicin-induced renal tubular epithelial cell injury, but the effect on the in vitro cultured cells is still rare. OBJECTIVE: To explore the mechanisms of basic fibroblast growth factor (bFGF) at different concentrations on preventing nephrotoxity mediated by genamicin on the primarily cultivated renal tubular epithelial cell models. METHODS: By use of enzyme and mesh screening, renal tubular epithelial cells were isolated from Kunming mice and purified, adjusting the cell concentration of 1×10~8/L, then cell suspension was moved to a 96-well culture plate and divided into different groups for culture: blank control group: normal culture; gentamicin group: 10, 30, 50 μL/hola (ie, 400, 1 200, 2 000 U/holes)arerecorded as G1, G2, G3; bFGF group: 20, 50, 80 μL/hole (ie, 90,225, 360 ng/hole) are recorded as B1, B2, B3; gentamicin plus bFGF intervention group: after adding bFGF 12 hours, then added gentamicin 12 hours, assigned into 9 dose subgroups, namely, G1B1, G1B2, G1B3, G2B1, G2B2, G2B3, G3B1, G3B2, G3B3, each subgroup contained four-hole complex. Cell morphology and quantity was observed. RESULTS AND CONCLUSION: Gentamicin showed a dose-dependent effect on the renal tubular epithelial cell injury, epithelial cells in the medium and high concentration groups exhibited shrinkage, rounded, swelling, poor adhesion, severely damaged cytoplasm and structural disorder. In the low concentration group, the number change of cells was not obvious, and fibroblasts began to appear; In the bFGF groups, cells were full, exhibited strong refraction, the cell number increased significantly, these manifestations were significant in 50 μL/hole concentration, and there was no significant difference compared with 80 μL/hore concentration; in case of gentamicin plus bFGF intervention, cells with low concentrations of gentamicin had no obvious damage to cells, which increased, the damaged cells collapse was reduced in the group of low concentration of gentamicin, cell shrinkage and poor adhesion were slightly relieved, high concentrations of bFGF intervention could yield to good cell morphology, but high concentrations of gentamicin caused cell swelling and necrosis of injury, which could not be improved by any concentrations of bFGF intervention. 50 μL/hole bFGF has antagonistic effect on the nephrotoxicity mediated by medium and low concentrations of gentamicin, but has no protection on high concentration of gentamicin-induced nephrotoxicity.

BACKGROUND: There is no effective method to identify liver cancer stem cells until now, which has become one of the major challenges in this field. OBJECTIVE: To explore a more effective and suitable way for the identification of liver cancer stem cells in hepatocellular carcinoma cell lines. METHODS: Firstly, the single cell separation technique was used to obtain single cells, which were seeded into 96-wel plates one by one. Secondly, cell sublines derived from single cell colonies (tumorigenic colonies) were selected and obtained. At last, the cells from these clones were transplanted into the forelimb armpits of nude mice to observe the tumorigenic ability. RESULTS AND CONCLUSION: The number of holes of single cells obtained was 371 by the single cell separation technique, and the success rate was 96.4%. According to the growth of cell clone, tumorigenic colonies were selected to be transplanted to the forelimb armpits of nude mice, and the tumor formation rate of the colonies was 100%. The identification model of single-cell-transplant-tumorigenic-test for liver cancer stem cells is confirmed to be preliminarily established, which lays the technology and methodological basis for the follow-up research.

Objective To explore the effect of acupuncture combined with Guishen Decoction (GD) on serum antisperm antibody (ASAb) level of rat models with immunological infertility. Methods Active immunization was used to establish rat models with positive serum ASAb. Eighty SD rats were alloc ated to four group s: Group A (treated with acupuncture and GD), Group B( treated with GD), Group C (treated with acupuncture), Group D (model control group). GD is composed of Ra dix Re hmanniae Praeparata, Semen Cuscutae, Fructus Ligustri Lucidi, Rhizoma Dioscorae, Fructus Lycii, Ramulus Loranthi, etc.. Results The negative rate of serum A SAb in Group A was significant higher than that in Group B and Group C (P

Fluorescence spectroscopy was used to investigate the efficiency of coupling peptides to gold nanoparticles via 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride-N-hydroxysuccinimide (EDC-NHS).The experiment conditions including buffer solution, pH value and concentrations of buffer solution, concentrations of NHS and EDC, concentration ratios of NHS to EDC, and coupling reaction time on the coupling efficiency were investigated and optimized.The experimental results indicated that the optimized experimental conditions were as follows: 25 mmol/L HEPES buffer solution, pH 7.0;2∶1 of concentration ratio of NHS to EDC, 0.4 mol/L NHS, 0.2 mol/L EDC, and coupling reaction time of 24 h.This study may provide references for the relative research involving coupling peptide or protein with gold nanoparticles

Objective An investigation of new cultivars of Amomum villosum Lour.(AVL) with high yield and good quality was carried out,thus to supply evidences for the identification of AVL cultivars in accordance with the morphological features of their flowers and fruits.Methods An investigation of AVL species from the genuine producing areas of Yangchun city of Guangdong province was performed.The morphological features of flowers and fruits of two cultivars(Changguo and Yuanguo) as well as one breeding type(Chunxuan type) were examined.Results Specific and significant features were screened out in different cultivars of AVL.Conclusion There exit specific features in flowers and fruits of different cultivars of AVL from Yangchun.

BACKGROUND:Until now, little has been reported on establishment of pancreatic stem cell strains and lines,and the purification of pancreatic stem cells is difficult since the cell line establish rate is low.OBJECTIVE:To explore a more rational and effective technique of in vitro separation and continuous passage of pancreatic stem cells, with the hope to establish cell strains and even cell lines and to lay the foundation for the follow-up study of pancreatic stem cells in the treatment of diabetes.METHODS:Firstly, Percoll discontinuous density gradient centrifugation method was applied to separate the mouse pancreatic endocrine portion from the exocrine portion, then to obtain cell strains with highly proliferative ability and low differentiation from pancreatic endocrine portion-the islet. We used mouse embryonic fibroblasts treated with mitomycin C as a feeder layer, for in vitro continuous culture of islet-derived pancreatic stem cells under feeder layer conditions until they were transferred to the 30th passage to establish cell lines. Then pancreatic stem cell line derived from pancreatic islet was detected and identified by a series of tests including growth characteristic test, morphological observation, related molecular marker identification and differentiation characteristic identification.RESULTS AND CONCLUSION:In the continuous process of passage, pancreatic stem cells showed active proliferative ability, and maintained the typical morphological characteristics of stem cells and expression of pancreatic stem cell marker-Nestin. After induction, pancreatic stem cells showed insulin gene expression,reflecting their differentiation potential. Therefore, under the condition of feeder layer, the pancreatic stem cell line derived from Kunming mice was successfully established and the related identification was completed,which lays the foundation for the following research.

This study sought to make a biomechanical analysis of the diffuse axonal injury(DAI) animal model caused by nonimpact with half bound head in cats. A three-dimensional finite element model of cat's head was established. The head of an anesthetized cat was scanned in 2 mm section. The nods and element meshes were signed out according to the geometry of every section. The geometric data were put into the computer and the element mesh body of cat's head was established in vizi CAD system. The maximum stress, minimum stress and von Mises stress were calculated by Super SAP (93ed) finite elemental software when the force was loaded on the right or left side of model in zero section. The analysis showed that the maximum stress appeared in the anterior and posterior loaded point and extended to cranial base in the cranial shell. There was high stress in the brain surface also. Because of cerebellar tentorium, cerebral falx, petrosal bone and sellar process, the stress did not decrease equivalently while approaching the deep brain, but it was distributed in cerebral-cerebellar peduncles, brain stem, corpus callosum and basal ganglia area at high values. The results suggest that the stress caused by rotational force is widespreadly and unequivalently distributed in brain tissue, which is mainly effected by the cerebellar tentorium, cerebral falx and the irregular geometric forms of cranial bone.
Animals ; Biomechanical Phenomena ; Brain ; pathology ; Brain Injuries ; pathology ; Cats ; Diffuse Axonal Injury ; pathology ; Finite Element Analysis ; Head ; Models, Animal ; Rotation ; Skull

BACKGROUND: There is a great debate but little research addressing the cell suspension obtained from the digested mantle tissues can effective amplify and form pearl sac in vitro, thus producing pearl. OBJECTIVE: To establish an effective technique and method of in vitro separation and culture of mantle of the pearl oyster (Pinctada martensii), and to determine the optimal method of forming pearl sac with the intact structure and secretion function, thus producing pearl. DESIGN, TIME AND SETTING: Single sample observation was performed at the School of Basic Medicine, Guangxi Traditional Chinese Medical University, between August and December in 2008. MATERIALS: Pearl oyster (Pinctada martensii) aged 1.0 2.0 years, were offered by Yingpan Pearl Industrial Co., Ltd. Of Beihai City, China; the self-modified marine shellfish balanced salt solution; the self-prepared concha pteriae serum and concha pteriae body fluid; keratinocyte growth factor was purchased from Sigma, USA. METHODS: The mantle of pearl oyster (Pinctada martensii) was digested with 2.5 g/L trypsin, the harvested cells were cultured using M199 medium containing 10% fetal bovine serum and supplemented with 10 μg/L keratinocyte growth factor, 10% self-prepared concha pteriae serum and concha pteriae body fluid. The cultivation was performed for 30 days. MAIN OUTCOME MEASURES: Cell growth characteristics and growth state. RESULTS: The pearl mantle epithelial cells cultured in vitro were shown to proliferate rapidly, secrete productively, and the muscle cells showed a great proliferation, finally encapsulated the mantle epithelial cells to form pear sac with the intact structure and strong secretion function. CONCLUSION: Using the modified culture technology and culture system, the addition of keratinocyte growth factor can obtain the well growing and secreting pearl sac during in vitro culture of mantle cells.

Chronic spontaneous urticaria is defined as recurrent wheals of unknown etiology for more than 6 weeks,with or without angioneurotic edema.The key step in the pathogenesis of chronic spontaneous urticaria is activation and degranulation of mast cells and basophils.Omalizumab,a recombinant humanized IgG monoclonal antibody,can selectively bond the Fc region of free IgE antibodies,and then block IgE-FCeR Ⅰ-mediated activation of mast cells and basophils.Three phase Ⅲ clinical trials have been reported on the efficacy of omalizumab in the treatment of chronic spontaneous urticaria,which confirm the efficacy and safety of omalizumab in the treatment of refractory chronic spontaneous urticaria.