AIMA method of coculture of brain capillary endothelial cells (BCECs) and astrocytes of rats was used to evaluate nanoparticle's blood-brain barrier (BBB) transcytosis and toxicity at the endothelial tight junction.

METHODSA lipophilic fluorescent probe, 6-coumarin, was incorporated in poly (ethyleneglycol)-poly (lactide) nanoparticle using double emulsion/solvent evaporation method. BCECs and astrocytes were firstly isolated from brain of newborn rats and characterized by their morphology and immunocytochemistry staining, separately. Subsequently, a coculture model with BCECs on the top of micro-porous membrane of cell culture insert and astrocytes on the bottom side was established. The permeability of 14C-labeled sucrose and nanoparticle were determined, separately.

RESULTSThe mean weight-based diameter of 6-coumarin loaded nanoparticles was (102.4 +/- 6.8) nm, with zeta potential of (-16.81 +/- 1.05) mV. BCECs were positive for factor VIII staining and glial fibrillary acidic protein was expressed in astrocytes. The transendothelial electrical resistance reached up to (313 +/- 23) omega x cm2. The tight junction between BCECs in the coculture model could be visualized by both scanning electron microscopy and transmission electron microscopy. The unchanged paracellular transport of sucrose proved that nanoparticle with concentration lower than 200 microg x mL(-1) did not impact the integrity of BBB endothelial tight junctions. The permeability of 10 microg x mL(-1) 6-coumarin labeled nanoparticle was 0.29 x 10(-3) cm x min(-1).

CONCLUSIONThis in vitro experimental model of rat BBB was close to resemble the in vivo situation for examination of the permeability of nanoparticle and toxicity evaluation.

Animals ; Animals, Newborn ; Astrocytes ; metabolism ; ultrastructure ; Biological Transport ; Blood-Brain Barrier ; Brain ; blood supply ; cytology ; Capillaries ; cytology ; Cell Membrane Permeability ; Coculture Techniques ; Coumarins ; administration & dosage ; pharmacokinetics ; toxicity ; Endothelial Cells ; metabolism ; ultrastructure ; Factor VIII ; metabolism ; Glial Fibrillary Acidic Protein ; metabolism ; Nanoparticles ; Polyesters ; Polyethylene Glycols ; Rats ; Rats, Sprague-Dawley ; Sucrose ; pharmacokinetics

OBJECTIVETo explore the influences of orthodontic tooth movement on estrous cycle and estrogen in female rats.

METHODSTwo hundred female rats were divided into control group, one-time activation group, four-time activation group and sham-operated group. Each group was divided again into 4 sub-groups according to the different stage of the estrous cycle. The force-loading groups received repeated intermittent force. Serum estrogen was measured and the change of estrous cycle was recorded.

RESULTSThere were significant variations in the estrogen level among the groups which received mechanical force during the same stage of the estrous cycle (P < 0.01) and among the subgroups within each group (P < 0.05). These experimental treatments affected the estrous cycle of the rats in the groups received force in metestrus and diestrus.

CONCLUSIONSEstrus of rats was the appropriate time for the orthodontic force.

Animals ; Estrogens ; blood ; Estrous Cycle ; physiology ; Female ; Rats ; Rats, Wistar ; Tooth Movement Techniques

OBJECTIVETo explore the effects of applying force during the different stages of estrous cycle on orthodontic tooth movement of rats, so as to offer an experimental princinple for women's orthodontic treatment.

METHODS80 female 3-month-old Wistar rats, which had a stable and five-day estrous cycle, were used. They were randomly divided into control groups and loading-force groups. Each group was divided again into 4 sub-groups according to the different stages of the estrous cycle. The loading-force groups received repeated intermittent orthodontic force for four times seperately during the same stage of the estrous cycle. The distance between upper incisor and the first molar on the left was measured. The data were analyzed by One-way ANOVA, S-N-K.

RESULTSThere were significant variations in the amount of tooth movement among the groups which received the mechanical force during the different stages of the estrous cycle (P <0.01). There were significant variations in the amount of tooth movement between the groups received force during pre-estrus and estrus (P <0.05). There were not significant variations between the groups received force during metestrus and diestrus (P > 0.05). The largest amount of tooth movement was in estrus group and smallest in pre-estrus one.

CONCLUSIONThe effect of tooth movement depended on which stage of estrous cycle was chosen to be the time of applying orthodontic force.

Analysis of Variance ; Animals ; Estrous Cycle ; Female ; Humans ; Incisor ; Molar ; Rats ; Rats, Wistar ; Tooth Movement Techniques

OBJECTIVETo investigate the value of computer-assisted slide-screening system (ThinPrep imaging system, TIS) in the diagnosis of cervical Thinprep smears.

METHODSA total of 19 600 ThinPrep smears were collected, including 9800 slides by TIS-assisted screening from September 2011 to March 2012 and 9800 slides by manual screening from September 2010 to April 2011 as control. The detection rates of abnormal cells and common microbial infection by the different screening methods were compared. With histopathological diagnosis of colposcopic biopsy as the gold standard, the screening efficiency and correlation of cytologic diagnosis among different screening methods were analyzed.

RESULTSCompared with manual screening, the detection rate of abnormal cells in 9800 cases by TIS-assisted screen was increased from 5.4% (525/9800) to 6.8% (665/9800), mainly in the categories of ASCUS and LSIL (P < 0.05). TIS had a higher accordance rate between cytologic diagnosis and histopathological diagnosis in the NILM and ASCUS than that by manual screening. False-negative rate of finding abnormal cells by TIS decreased from 8.5% (17/200) to 0.7% (2/289, P < 0.01) with an increased sensitivity compared to manual screening, although the specificity was similar. Both TIS and manual screening had advantages and disadvantages respectively in the detection of microbial organisms. TIS improved screening efficiency by 50%.

CONCLUSIONTIS improves not only the screening efficiency but also the detection of abnormal cells with a reduced false negativity, and it therefore has a broad application prospect.

Adenocarcinoma ; diagnosis ; pathology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Candida ; isolation & purification ; Carcinoma, Squamous Cell ; diagnosis ; pathology ; Cervical Intraepithelial Neoplasia ; diagnosis ; pathology ; Cytodiagnosis ; False Negative Reactions ; Female ; Humans ; Image Processing, Computer-Assisted ; instrumentation ; Mass Screening ; Middle Aged ; Sensitivity and Specificity ; Trichomonas vaginalis ; isolation & purification ; Uterine Cervical Dysplasia ; diagnosis ; pathology ; Uterine Cervical Neoplasms ; diagnosis ; pathology ; Vaginal Smears ; Young Adult

A typical chronic myeloid leukaemia (aCML), which shows both myeloproliferative and myelodysplastic features, is a type of myeloproliferative/myelodysplastic disease as defined by the World Health Organisation (WHO) classification of the myeloid neoplasms. Because of the presence of neutrophilic leukocytosis, aCML may resemble chronic myelogenous leukemia (CML). However, in contrast with CML, aCML does not have the Philadelphia chromosome or the bcr/abl fusion gene. With the continuous karotype analysis of aCML, several changes in the karyotype of aCML have been detected. However, few are recurring and no specific cytogenetic changes have been associated with aCML. Nonspecific cytogenetic abnormalities can be observed in 56%~82% of aCML cases. Although the most frequent abnormalities include trisomy 8 and del (20q), abnormalities involving other chromosomes such as 12, 13, 14, 17, and 19 have also been described. In this report we describe a case of aCML with trisomy 13.
Adult ; Chromosomes, Human, Pair 13 ; Female ; Humans ; Leukemia, Myeloid, Chronic, Atypical, BCR-ABL Negative ; genetics ; Trisomy

BACKGROUNDSkin lesions are common manifestations in systemic lupus erythematosus (SLE). It is still unknown what the definite pathogenesis of skin involvement was and whether DNA participated in it. Our study was designed to explore the pathogenetic role and nature of nuclear antigen (DNA) deposited in the skin lesions of patients with SLE.

METHODSThirty skin samples from patients with SLE and 2 normal skin samples were studied. Extracellular DNA was evaluated by indirect immunofluorescence methods. The deposited immune complexes were extracted by cryoprecipitation, and DNA was then isolated with phenol and chloroform. DNA fragment sizes were detected by agarose gel electrophoresis. Finally, 8 different probes were used to analyze the origin of these DNA molecules using Dot hybridization.

RESULTSExtracellular DNA staining was found only in skin lesions, mainly those located in the basement membrane zone, vascular wall, and hair follicle wall. Normal skin and non-lesion SLE skin showed no fluorescence at locations outside the nuclei. There were no differences in the rate and intensity of extracellular DNA staining when comparing active phase to remission phase patients. No relationship was found between extracellular DNA and circulating anti-dsDNA antibodies. Deposited DNA fragments clustered into four bands of somewhat discrete sizes: 20 000 bp, 1300 bp, 800-900 bp, 100-200 bp. Small sized fragments (100-200 bp) were positively correlated with disease activity (P < 0.05, r = 0.407). Dot hybridization showed significant homology of the various extracellular DNA fragments examined with human genomic DNA, but not with DNA from the microorganisms and viruses we examined. There were also homologies between DNA samples from different individuals.

CONCLUSIONSDNA and its immune complexes may contribute to the pathogenesis of skin lesions in SLE. These DNA molecules range in size from 100 bp to 20 kb and may be endogenous in origin.

Antibodies, Antinuclear ; blood ; Antigen-Antibody Complex ; analysis ; DNA ; analysis ; immunology ; Humans ; Lupus Erythematosus, Systemic ; immunology ; Skin ; immunology ; Staining and Labeling

Objective To establish simple,stable and reliable rat models of oxygen glucose deprivation/reoxgenation(OGD/R)in adult neural stem cells(NSCs)in vitro.Methods The NSCs from adult Fisher344 rats were cultured in serum-free medium and identified using nestin and DAPI immunofluorescent double staining.These cells were washed with a Earle′s balanced salt solution without glucose for 2 times,then,incubated for different periods(2,4,6,8 and 10 h)in a trigas incubator with an atmosphere of 1％ O2,5％CO2 and 94％ N2,98％ humidity at 37 ° C.And then,these cells were removed from the anaerobic incubator,washed,and added DEME/F12 containing bFGF supplement.A normoxic-normoglycemic control group was employed.Morphological assessment of NSCs was performed by light microscopy after re-oxgenation for 24 h; CCK-8 colorimetric method was used to determine the survival and proliferation of NSCs,and flow cytometry was employed to detect the apoptosis of NSCs.Results After the setting of oxygen glucose deprivation for 2 h,the OD value and the survival rate in the OGD cells were increased as compared with those in control group without significant difference(P＞0.05).While the morphological damage of NSCs aggravated gradually and the OD value decreased in OGD cells following the prolongation of times; under the setting of oxygen glucose deprivation for 6 h,the OD value in OGD cells obviously decreased as compared with that in the control group(P＜0.05); under the setting of oxygen glucose deprivation for 6 h,the survival rate obviously decreased and the apoptosis rate significantly increased in OGD cells as compared with that in the control group(P＜0.05); under the setting of oxygen glucose deprivation for 6 h,the apoptosis rate of NSCs excessed to 50％.Conclusion By means oftrigas incubator,simple,stable and reliable models of OGD/R in NSCs in vitro can be successfully established.

This paper discusses the application of virtual reality technology in the 3-D visible human body and acupuncture research. Based on the 3-D visible human fused with the localization information and hierarchy of acupoints, the paper analyzes the force against the needle and haptic rendering during the needle manipulation according to the physical properties of different tissues. A haptic model is constructed to demonstrate the force behaviors during acupuncture, and the force will be produced and passed to the manipulator by a force feedback device. It enriches the contents of 3-D visible human project, provides a dynamic simulation instrument for acupuncture teaching, and supplies a platform for acupuncture research.
Acupuncture Therapy ; methods ; China ; Computer Simulation ; Feedback ; Humans ; Imaging, Three-Dimensional ; User-Computer Interface ; Visible Human Projects

OBJECTIVESTo investigate whether hepatitis B virus X gene alone is sufficient to transform the non-transformed immortalized human liver cell line QSG7701 and induce hepatocellular carcinoma in vivo.

METHODSpCMVX/QSG7701 cells were transplanted into subcutaneous tissue of nude mice. pRcCMV2/QSG7701 and QSG7701 cells were used as control. Tumor formation was checked within 5 weeks after transplantation. The activity and food intake of the nude mice were recorded. The texture, volume and metastasis of transplantation tumor were observed grossly, and the HE stained transplantation tumor tissues were observed under optical microscope.

RESULTSThe transplantation tumor occurred in all of the six nude mice inoculated with pCMVX/QSG7701 cells at the second week after inoculation. No metastatic tumor was found in other organs. Transplant tumor was not formed in all of the negative control groups. The activity, eating and drinking of the nude mice transplanted with pCMVX/QSG7701 cells were normal, while their weights were increased gradually in the first 3 weeks. Since the 4th week after transplantation with pCMVX/QSG7701 cells, the activity of the mice was decreased and their body weight was no longer increased. It is interesting that the mental state and eating of those nude mice inoculated with pRcCMV2/QSG7701and QSG7701 cells were normal, and the weight was increasing all the time after inoculation. HE staining analysis confirmed that the transplanted tumor was hepatocellular carcinoma.

CONCLUSIONHBx alone is sufficient to transform the non-transformed immortalized human liver cell line QSG7701 and induce hepatocellular carcinoma in vivo.

Animals ; Carcinoma, Hepatocellular ; Hepatitis B virus ; genetics ; Hepatocytes ; Humans ; Liver Neoplasms ; Mice ; Mice, Inbred BALB C ; Mice, Nude

OBJECTIVETo investigate the effects of astragalus on tubulointerstitial lesions in rats with IgA nephropathy (IgAN) and to explore the possible mechanism.

METHODSTwenty-eight Sprague-Dawley rats were randomly assigned to three groups. The rat model of IgA nephropathy was induced by intragastric administration of bovine serum albumin and injections of LPS and CC14. Six weeks later, the rats with IgAN were randomly treated with oral astragalus (3 g/kg/d, for 6 weeks) or normal saline. Normal control rats which were not subjected to IgAN were treated with normal saline. The number of urinary erythrocytes and urinary protein and B-D-N-Acetyl glucosaminidase (NAG) contents were determined by Pan-automatic biochemistry analyzing meter. Expression of monocyte chemotactic protein-1 (MCP-1) and nuclear factor-kappa B (NF-kappaB) in tubulointerstitial tissues were analyzed by immunohistochemistry. A semiquantitative score was used to evaluate the degree of renal pathologic lesions.

RESULTSThe number of urinary erythrocytes (74.02+/-16.58 / microL vs 383.23+/-4.94 /microL) and urinary protein (13.88+/-4.94 vs 59.82+/-14.73 mg/L) and NAG contents (2.84+/-0.31 vs 5.24+/-0.80 U/L) in the astragalus-treated IgAN rats decreased remarkably compared with those in the IgAN rats without astragalus treatment (P<0.01). Expression of the NF-kappaB and MCP-1 in the renal tissues in the IgAN rats without astragalus treatment was significantly higher than that in the astragalus-treated IgAN rats and normal control rats (P<0.01). There were significant differences in the scores of renal pathologic lesions between the IgAN rats with or without astragalus treatment (6.03+/-0.46 vs 10.57+/-1.23; P<0.01).

CONCLUSIONSAstragalus can decrease the number of urinary erythrocytes and urinary protein and NAG contents, and relieves tubulointerstitial lesions, possibly through the down-regulation of NF-kappaB and MCP-1 expression in rats with IgAN.

Animals ; Astragalus Plant ; Chemokine CCL2 ; analysis ; Glomerulonephritis, IGA ; drug therapy ; metabolism ; pathology ; Immunohistochemistry ; Kidney Tubules ; pathology ; Rats ; Rats, Sprague-Dawley ; Transcription Factor RelA ; analysis