Objective To investigate the proliferation of rat bone marrow endothelial progenitor cells (EPCs) in the culture under different concentrations of TGF-?1 in order to optimize the culture conditions. Methods EPCs harvested from bone marrow by flushing fresh rat femur and tibia were isolated by density gradient centrifugation. The isolated cells were further purified and enriched by fast adhere to fibronectin coated dish. Flow cytometry was applied to enrich the cells positive to CD34, CD133 and VEGFR-2. EPCs were expanded in M199 medium in presence of different concentrations of TGF-?1 (10-300 pg/ml). Total cell output was recorded; and the expressions of CD34, CD133, and VEGFR-2 at different cell passages were analyzed through flow cytometer. Results Fast adhere cell group showed significantly higher positive proportion of CD34, CD133, and VEGFR-2 than unsorted cells (P

Objective To identify and analyze the species of vaginal lactobacilli between patients with bacterial vaginosis (BV) and healthy women at childbearing age in Inner Mongolia. Methods From Jun. 2008 to Dec. 2008, 203 Mongolian healthy women, 74 Han healthy women and 102 Mongolian patients with BV from 3 pastoral areas were enrolled in this study. Isolation and culture of lactobacilli from vaginal wall were performed by modified culture medium. DNA of lactobacilli were extracted and sequenced. H2O2 were detected by TMB-HRP-MRS. Results(1)The rate of lactobacilli identification were 76.8%(156/203) in Mongolian healthy women and 21.6% (22/102) in Mongolian patients with BV, which reached statistical difference(P＜0.01).Lactobacilli identification in Han healthy women [82.4%(61/74)] did not show significant difference with that of Mongolian healthy women (P＞0.05). (2) The total of 193 strains and 11 species of Lactobacillus were detected in 203 Mongolian healthy women. Meanwhile,22 strains and 4 species of Lactobacillus were found in 102 Mongolian BV cases.(3)The rate of H2O2 generating Lactobacilli was 27.3% (6/22) in Mongolian BV patients and 75.7% (56/74)in Mongolian healthy women, which showed statistical difference(P＜0.05). Conclusions The rate of Lactobacillus was not related with the race of women in pastoral area in Inner Mongolian. The amount of lactobacilli and H2O2 generating Lactobacilli in the vagina of BV patients was remarkably lower than those of healthy women at childbearing age.

OBJECTIVETo investigate the correlation of the deleted azoospermia (DAZ) gene copy related to gr/gr and b2/b3 deletions in the AZFc region with male spermatogenic impairment.

METHODSThis study included 121 infertile men with different de- grees of spermatogenic impairment and 95 healthy donors from the sperm bank. Using PCR, PCR-RFLP, and Y chromosome specific sequence tagged sites (STS) , we analyzed the association of DAZ gene copy deletions related to gr/gr and b2/b3 deletions in the AZFc region with spermatogenic impairment.

RESULTSThere were 15 cases of gr/gr deletion (12. 40% ) and 6 cases of b2/b3 deletion (4.96%) in the infertility group as compared with 13 cases of gr/gr deletion (13.68%) and 1 case of b2/b3 deletion (1.05%) in the control. Analysis of the DAZ-specific single nucleotide variant (SNV) loci revealed 11 gr/gr-DAZI/DAZ2 deletions (9.09%), 4 gr/gr-DAZ3/DAZ4 deletions (3.31%), and 6 b2/b3-DAZ1/DAZ2 deletions (4.96%) in the infertile men in comparison with 3 gr/ gr-DAZ1/DAZ2 deletions (3.16%), 10 gr/gr-DAZ3/DAZ4 deletions (10.53%), and 1 b2/b3- DAZ3/DAZ4 deletion (1.05%) in the control.

CONCLUSIONPartial deletions of gr/gr and b2/b3 exist in both healthy men and male patients with different degrees of spermatogenic impairment and cannot be considered as a risk factor for spermatogenesis impairment. However, deletions of different DAZ duplicons in gr/gr and b2/b3 deletions have different effects on spermatogenesis. DAZ1/DAZ2 instead of DAZ3/DAZ4 deletions might be associated with spermatogenesis impairment.

Prostate cancer (PCa) is one of the most common malignant tumors as well as a frequent cause of cancer-related mortality in men worldwide. The test of serum markers has dramatically improved the early diagnosis of PCa, but its underlying molecular mechanisms are not yet completely identified. Long noncoding RNA (IncRNA) is emerging as a new player in the PCa paradigm demonstrating its potential roles in both oncogenic and tumor suppressive pathways. LncRNA is frequently aberrantly expressed in the majority of PCa cases. This review highlights recent findings of the aberrant expression of lncRNA in PCa and discusses its novel roles in the diagnosis, prediction, prognosis, metastasis, and potential clinical treatment of PCa.
Humans ; Male ; Prognosis ; Prostatic Neoplasms ; metabolism ; RNA, Long Noncoding ; metabolism

Wuzhuyu Tang is a classical formula for treating migraine, but its' pharmacological ingredients is unclear yet. Present study employed the everted intestinal sac model to collect the absorption samples of 10 kinds of Wuzhuyu decoction, and then analyzed the contents of 9 ingredients in Wuzhuyu Tang and absorption samples quantitatively or semi-quantitatively by HPLC-DAD method. Reserpine was used to establish the mice model of migraine, and then the contents and activities of 5-hydroxytryptamine, noradrenaline, dopamine, nitric oxide and nitricoxide synthase in brain tissues and serums were determined respectively after oral administration of Wuzhuyu Tang. Using the partial least squares regression method to correlate the total absorption quantity of 9 ingredients and pharmacodynamics. The result shows that limocitrin-3-O-beta-D-glucoside, ginsenoside Rg1 and Rb1, rutaevine, limonin, evodiamine and rutaecarpine are the main ingredients influenced the effects in absorption samples in everted intestinal sacs, especially ginsenoside Rg1, rutaevine, evodiamine and rutaecarpine among them have obvious improving effects to most pharmacodynamics index, might be the pharmacological ingredients influenced the therapeutical effects of Wuzhuyu Tang treating migraine.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; Male ; Mice ; Mice, Inbred ICR ; Migraine Disorders ; drug therapy ; Rats ; Rats, Wistar

Objective To study the difference of gene expression profile between the radioresistant human nasopharyngeal carcinoma cell line CNE-2R and CNE-2,and to screen the signaling pathway associated with radioresistance of nasopharyngeal carcinoma.Methods The radioresistant nasopharyngeal carcinoma cell line CNE-2R was constructed from the original cell line CNE-2.CNE-2R and CNE-2 cells were cultured and administered with 60Co γ-ray irradiation at the dose of 400 cGy for 15 times.Human-6v 3.0 whole genome expression profile was used to screen the differentially expressed genes.Bioinformatic analysis was used to identify the pathways related to radioresistance.Results The number of the differentially expressed genes that were found in these 2 experiments was 374.The Kegg pathway and Biocarta pathway analysis of the differentially expressed genes showed the biological importance of Toll-like receptor signaling pathway and IL-1 R-mediated signal transduction pathway to the radioresistance of the CNE-2R cells and the significant differences of 13 genes in these 2 pathways,including JUN,MYD88,CCL5,CXCL10,STAT1,LY96,FOS,CCL3,IL-6,IL-8,IL-1α,IL-1B,and IRAK2(t=13.47-66.57,P<0.05).Conclusions Toll-like receptor signaling pathway and IL-1R-mediated signal transduction pathway might be related to the occurrence of radioresistance.

BACKGROUND:The molecular mechanism of traumatic deep vein thrombosis is complex.Numerous studies focus on clinical observation and epidemiology,but its molecular mechanism has not been a new breakthrough.OBJECTIVE:By use of gene array technology,this study was aimed to study the expression changes of matrix metalloproteinases in rat models of traumatic deep vein thrombosis,and to explore the roles of matrix metalloproteinases in traumatic deep venous thrombosis.METHODS:A total of 150 SD rats,SPF grade,of 8-12 weeks old,body weight of 250-300 g,were divided at random into normal control group (n=10) and model group (n=140).Rat traumatic deep venous thrombosis models were set up by clamping the femoral vein and fixing the bilateral hind limbs,and the fixation of hip spica with plaster bandage was conducted in each group.Then rats were divided into 7 subgroups:post-traumatic 0.5 hours,post-traumatic 2.5 hours (initial period of thrombosis),post-traumatic 25 hours (thrombogenesis at thrombotic crest-time),post-traumatic 25 hours non-thrombogenesis at the thrombotic crest-time),post-traumatic 72 hours (thrombus resolution),post-traumatic 72 hours thrombus insolution) and post-traumatic 168 hours (nonthrombosis).At the corresponding phasess,the femoral vein tissues were incised,and total RNA of femoral vein was extracted using Trizol one-step method.Applying Genechip Rat Genome 430 2.0 genechips,the gene expressions in femoral vein were detected in different groups.The rate of traumatic deep venous thrombogenesis and non-thrombogenesis,the rate of thrombi solution and insolution were observed;the expressions of matrix metalloproteinases and tissue inhibitor of metalloproteinases at different time phases was detected by gene array data analysis.RESULTS AND CONCLUSION:Three model rats died and the remaining 147 rats were involved in the final analysis.At the post-traumatic 25 hours,the rate of thrombogenesis was 50.5% and nonthrombogenesis was 49.5%.To the post-traumatic 168 hours,the rate of thrombus solution was 56.7% and thrombus insolution was 43.3%.Both matrix metalloproteinases and tissue inhibitor of metalloproteinases exhibited differential expressions in the course of traumatic deep venous thrombosis.Under the thrombus insolution state,matrix metalloproteinases continued to show a high expression,tissue inhibitor of metalloproteinase expression was down-regulated in the thrombus formation,was significantly inhibited in the thrombus insoluUon process.In the process of traumatic deep vein thrombosis and insolution,matrix metalloproteinase was closely related to traumatic deep vein thrombosis,the matrix metalloproteinase/tissue inhibitor of metalloproteinases are likely to affect the biological state of thrombosis.

Background Application of confocal microscopy in the investigation of ocular surface system in living eye have been greatly extended in two decades.In vivo confocal microscopy allows the evaluation of the normal or pathological tissue at the cellular level.However,seldom study about the morphology of normal in vivo human bulbar conjunetiva under the confocal microscopy was induced. Objective Present study was to analyze the appearance of normal bulbar conjunetiva cells under the in vivo confocal microscopy. Methods Laser scanning confocal microscopy was used to examine the morphological characteristics of the bulbar conjunetiva in 21 eyes of 15 healthy volunteers.The parameters of confocal microscopy were as follows:resolution 1 μm,wave length 670 nm,field range 400 μm×400 μm.The epithelial cell numbers,dendritics cells density and goblet cells density from superior,inferonasal,nasal and temporal bulbar conjunctiva were calculated respectively and compared and imaged by the HRT3 Rostock Cornea Modual. Results The superficial epithelial cells of the bulbar conjunctiva was seen with the small cell nuclei and blur border.The borders of basal epithelial celt were clearly visible without cell nuclei.The presumed goblet cell presented with a large hyperreflective oval-shaped cell with relatively homogeneous brightness,crowed in groups or mainly dispersed.The orfices at the epithelial surface represented the goblet cells,showing some open and expel contents.The dendritic cell appeared to be hyperreflective corpuscular particles with visible processes among conjunetival epithelial cells.A few dense white fiber meshwork was exhibited in conjunctival stroma with the traverse blood vessels containing cellular elements.The superficial and basal epithelial cell densities were 2556±692and 2985±376 cells/mm2 respectively,and overall goblet and mature dendritic cells densities were 77±39 and 26±35 cells/mm2 respectively.Significant differences were proved in globet cell density and dendritic density among different conjunetival zones(P=0.001,P=0. 000),however,the alteration of conjunetival epithelium cells was insignificant in different area(P=0.204,P=0.130).Conclusion Confocal microscopy is a useful tool for the study of morphology of human bulbar conjunctiva cells in vivo.which offer a valuable aid in the diagnosis of ocular surface disease.

Objective To observe the effects of electro-acupuncture on the E-selectin and resistin protein expression in the hippocampus of rats of chronic stress depression;To explore the role and mechanism of electro-acupuncture in the micro-environment of brain with depression. Methods 40 Fourty rats were randomly divided into four groups:blank group, model group, model+EA group, and model+fluoxetine group by using chronic stress combined with solitary raising methods, 10 rats in each group. Electro-acupuncture intervention was used 1 hour before the modeling. Electro-acupuncture was given at points Baihui (Du 20) and Yintang (Extra) for twenty minutes (2 Hz, 0.6 mA) every day. Fluoxetine was given at the volume of 5 mL/kg, dosage of 10 mg/kg. E-selectin and resistin protein expression in hippocampus were determined by biotin label-based antibody array. Results Compared with the blank group, the protein expression of E-selectin and resistin in the model group increased (fold change=1.23, 1.22). Compared with the model group, E-selectin and resistin expression decreased (fold change=0.65, 0.62;fold change=0.76, 0.65). Conclusion Electro-acupuncture intervention could down-regulate E-selectin and resistin expression in the hippocampus of chronic stress depression model.

Objective To explore the variation trends of interleukin-1β( IL-1β) and intercellular adhesion molecule-1 (ICAM-1) in both normal and affected sides of brain tissues in rats with ischemia-reperfusion injury and the therapeutic action of electroacupuncture.Methods The cerebral ischemia-reperfusion model was established with suture embolization in the right middle cerebral artery.The rats were randomly divided into control group, model group and electroacupunture group.Each group was then divided into six subgroups by the time after operation (12 h,24 h,48 h,72 h,96 h,144 h), ten rats in each subgroup. Frozen sections of brain tissues were prepared and the expression of IL-1βand ICAM-1 in brain tissues of both sides were detec-ted by immunohistochemistry.Results The expressions of IL-1βand ICAM-1 showed typical bimodal pattern in both affected is-chemic region and contralateral normal region.In the model group, the peaks of IL-1βin the cerebral ischemic region were at 12 h and 48 h, while in the contralateral normal region the peaks were at 12 h and 144 h, the expression of IL-1βin the ischemic region was significantly higher than that in the contralateral normal region at 48 h (P<0.05), and lower at 96 h and 144 h (P <0.05).In the electroacupuncture group, the expressions of IL-1βin the ipsilateral region were significantly lower than that in the contralateral region at 24 h, 48 h and 144 h (P<0.05).In the model group, the peaks of ICAM-1 in the cerebral ischemic regions were at 24 h and 72 h, while in the contralateral normal regions the peaks were at 24 h and 144 h.In the electroacupunc-ture group, the expressions of ICAM-1 in the ischemic regions were significantly lower than that in the contralateral normal re-gions at all the 12 h, 24 h, 48 h, 72 h and 144 h (P<0.05).Conclusions Our findings suggest that electroacupuncture may inhibit the inflammation of ischemia/reperfusion brain tissue through reducing the expression of IL-1βand ICAM-1 to relieve the cerebral ischemia-reperfusion injury.