BACKGROUND: The effect of pituitary adenylate cyclase activating polypeptide (PACAP) during traumatic brain injury (TBI) and whether it can modulate secondary injury has not been reported previously. The present study evaluated the potential protective effects of ventricular infusion of PACAP in a rat model of TBI. METHODS: Male Sprague Dawley rats were randomly divided into 3 treatment groups (n=6, each): sham-operated, vehicle (normal saline)+TBI, and PACAP+TBI. Normal saline or PACAP (1g/5L) was administered intracerebroventricularly 20 minutes before TBI. Right parietal cortical contusion was produced via a weight-dropping method. Brains were extracted 24 hours after trauma. Histological changes in brains were examined by HE staining. The numbers of CD4+ and CD8+ T cells in blood and the spleen were detected via flow cytometry. RESULTS: In injured brain regions, edema, hemorrhage, inflammatory cell infiltration, and swollen and degenerated neurons were observed under a light microscope, and the neurons were disorderly arrayed in the hippocampi. Compared to the sham group, average CD4+ CD8– lymphocyte counts in blood and the spleen were significantly decreased in rats that received TBI+vehicle, and CD4– CD8+ were increased. In rats administered PACAP prior to TBI, damage was attenuated as evidenced by significantly increased CD4+, and decreased CD8+, T lymphocytes in blood and the spleen. CONCLUSION: Pretreatment with PACAP may protect against TBI by influencing periphery T cellular immune function.

By means of comparing biomasses of biodegradation fungi,Fusarium sp.HG-P-01 for ?-cypermethrin,a synthetic pyrethroid insecticide used widely in China,in five different media,the Czapek-Dox medium was selected as the best medium for mycelia growth.Furthermore,an experiment of central composite rotatable design(CCRD) was used to optimize the content of nutrient components.The optimal composition of C,N and P in media for HG-P-01 were 20.94 g/L,1.82 g/L and 1.66 g/L,respec-tively,in which an expectant or real rate of ?-cypermethrin-degradation got to 96.34% or 93.78% by HPLC for a concentration of 50 mg/L after 24 h treatment.The predicted value in degradation rate model was con-sistent with that from HPLC method.

Objective to investigate the value of diffusion-weighted echo-planar MR imaging in the diagnosis of head and neck lesions.Methods Fifty-seven patients with 85 head and neck lesions were enrolled in the study,including 22 patients with 22 malignant tumors,13 patients with 13 benign tumors, 13 patients with 17 cystic and liquefactive lesions(including 8 patients with 12 cystic lesions,4 patients with 4 tumor necrosis,1 patients with 1 abcess)and 33 lymph nodes.The lesions were all confirmed by operation and clinical follow up.Echo-planar difffusion-weighted imaging (DWI)was performed with different b values (0,500,and 1,000s?mm~(-2)),and the apparent diffusion coefficients (ADCs)were measured.Results Malignant and benign tumors had different characteristics in DWI with different b values.With the increase of b value,the signal intensity of tumor/spinal cord ratio decreased quickly in DWI in benign tumors,while the signal intensity of tumor/spinal cord ratio remained similar in DWI in malignant tumors.The mean ADC value of'malignant tumors[(0.78?0.24)?10~(-3)mm~2? s~(-1)] was significantly lower than that of benign tumors [(1.48?+0.20)?10~(-3)mm~2?s~(-1)] (t = 8.9,P

Background The construction of tissue-engineered corneal endothelium needs the functional seeding cells,so how to culture a large amount of functional corneal endothelial cells (CECs) is an urgent problem to be solved.Objective The aim of this study was to evaluate the role of aqueous humor on bovine CECs in vitro.Methods Aqueous humor of 1.2 ml was collected from the anterior chamber of bovine and sterilized,and the liquid supernatant was obtained.The bovine CECs were isolated from bovine cornea and then cultured in low glucose Dulbecco Modified Eagle Medium with 10％ fetal bovine serum (FBS) in vitro.Aqueous humor was added into the medium with the final concentration of 2.5％,5.0％,l0.0％,15.0％ and 20.0％,respectively,and no aqueous humor was added in the control group.Cell counting kit-8 (CCK-8) assay was used to detect the absorbency value of CECs for the evaluation of cell proliferation.Progression of the cell cycle was analyzed by flow cytometry (FCM).After confluence of the cells was reached,1 ml plastic spear tip was used to scratch the cell single layer,and the cells were incubated consequently in medium with 10％ FBS and with or without aqueous humor for 24 hours.Healing area of the cell single layer was measured.The cells were incubated at a density of 6 × 105 cells/ml and cultured using medium with or without 10.0％ aqueous human for 5 days,and the number of the cells was analyzed by DAPI fluorescence technique.Results Under the phase-contrast microscopy,the confluent CECs showed a slabstone-like and hexagonal appearance.CCK-8 assay revealed that the absorbance values of CECs was significantly different among the various culture groups (F=4.051,P =0.007),and the absorbance value in different concentrations of aqueous human culture groups was significantly higher than that in the control group (P ＜ 0.01).FCM showed that the percentage of the cells in S-G2 phases was (34.80-±3.13)％ in the 10.0％ aqueous humors group and (23.06±1.13)％ in the control group,showing a significant difference (t =-5.729,P=0.005).The scratch test showed that the healing area of the cell signal layer was (0.116±0.019) mm2 in the 10.0％ aqueous humors group and (0.358 ±0.049) mm2 in the control group,showing a significant difference (t =13.842,P =0.000).The density of cells in the 10.0％ aqueous humor group was (1439± 1 10)/field,which was more than (1162±45)/field in the control group (t =-11.020,P=0.000).Conclusions Aqueous humor at the concentration of 10.0％ promote the growth and proliferation of bovine CECs.The result suggests that 10.0％ aqueous humor can be used as a promoting agent during the culture of CECs.

BACKGROUND: Long noncoding RNAs (lncRNAs) play pivotal roles in various malignant tumors. Epidermal growth factor receptor (EGFR) signaling is associated with the pathogenesis of cutaneous squamous cell carcinoma (cSCC). This study aimed to explore the role of LINC00520 in the development of cSCC via EGFR and phosphoinositide 3-kinase-protein kinase B (PI3K/Akt) signaling pathways. METHODS: A microarray analysis was applied to screen differentially expressed lncRNAs in cSCC samples. The A431 cSCC cell line was transfected and assigned different groups. The expression patterns of LINC00520, EGFR, and intermediates in the PI3K/Akt pathway were characterized using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blotting analysis. Cell proliferation, migration, and invasion were detected using the MTT assay, scratch test, and Transwell assay, respectively. Cell-based experiments and a tumorigenicity assay were conducted to assess the effect of LINC00520 on cSCC progression. This study was ended in September 2017. Comparisons between two groups were analyzed with t-test and comparisons among multiple groups were analyzed using one-way analysis of variance. The nonparametric Wilcoxon rank sum test was used to analyze skewed data. The enumerated data were analyzed using the chi-square test or Fisher exact test. RESULTS: Data from chip GSE66359 revealed depletion of LINC00520 in cSCC. Cells transfected with LINC00520 vector and LINC00520 vector + si-EGFR showed elevated LINC00520 level but decreased levels of the EGFR, PI3K, AKT, VEGF, MMP-2 and MMP-9 mRNAs and proteins, and inhibition of the growth, migration and adhesion of cSCC cells, while the si-LINC00520 group showed opposite trends (all P < 0.05). Compared with the LINC00520 vector group, the LINC00520 vector + si-EGFR group showed decreased levels of the EGFR, PI3K, AKT, VEGF, MMP-2 and MMP-9 mRNAs and proteins, and inhibition of the growth, migration and adhesion of cSCC cells, while the LINC00520 vector + EGFR vector group showed opposite results (all P < 0.05). CONCLUSION Based on our results, LINC00520-targeted EGFR inhibition might result in the inactivation of the PI3K/Akt pathway, thus inhibiting cSCC development.
Animals ; Carcinoma, Squamous Cell ; pathology ; prevention & control ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Disease Progression ; ErbB Receptors ; antagonists & inhibitors ; Female ; Humans ; Lymphatic Metastasis ; Mice ; Neoplasm Invasiveness ; Phosphatidylinositol 3-Kinases ; physiology ; Proto-Oncogene Proteins c-akt ; physiology ; RNA, Long Noncoding ; physiology ; Signal Transduction ; physiology ; Skin Neoplasms ; pathology ; prevention & control

OBJECTIVE: To establish a multiplexing Y-STR system and study haplotype frequencies of 4 Y-specific loci in China Han population. METHODS: DYS439, DYS390, GATA-A7.2 and DYS393 loci were amplified simultaneously and were analyzed by polyacrylamide gel electrophoresis and silver staining. RESULTS: When 558 unrelated male individuals from the Han population in China were tested by the multiplex system, DYS439, DYS390, GATA-A7.2 and DYS393 show 7,7,7,6 alleles, respectively. 180 different haplotypes were detected. The power of discrimination of this system was 0.9853. CONCLUSION The multiplex amplified system of these 4 Y-specific loci and their database are useful for human origin exploration and forensic practice.
Alleles ; China ; Chromosomes, Human, Y ; DNA/isolation & purification* ; Electrophoresis, Polyacrylamide Gel ; Ethnicity/genetics* ; Female ; Forensic Medicine ; Gene Frequency ; Haplotypes ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Silver Staining ; Tandem Repeat Sequences/genetics*

AIMTo introduce the manufacture and use of a simple, new type of drop recorder of frog heart.

METHODSTo improve the perfusion device of (see text for symbol) and Straub method. Two electrodes of drop recorder were fixed in an injector of 20 ml. The input tube, output tube and resistance tube were all made of plastic material.

RESULTSThis device could be used to observe effects of preload, after-load, hormone and electrolyte on the cardiac output in isolated frog heart.

CONCLUSIONThe new type of drop recorder was economical and could be easily operated, it could be also connected to computer. Using the new type of drop recorder, effects of various physical and chemical factors on cardiac function could be observed directly, accurately.

BACKGROUND: Endothelial progenitor calls are the cells that can form new blood vessels in the way of angiogenesis in the body,which updates the conventional theory of angiogenesis, vascular damage and repair after birth and provides new ideas for research and treatment of ischemic diseases.OBJECTIVE: To investigate the effects of dog umbilical cord blood endothelial progenitor call (UCB-EPC) transplantation on angiogenesis after myocardial infarction.DESIGN, TIME AND SETTING: An in vivo cytological experiment was performed at the Laboratory Center of Xinhua Hospital between May 2006 and March 2007.MATERIALS: One full-term pregnant hybrid dog was included for preparation of UCB-EPCs. Thirty-six adult dogs were randomly divided into a cell transplantation group (n = 18) and a model control group (n = 18).METHODS: Acute myocardial infarction model was established in each group by ligation of antedor descending coronary artery.In the cell transplantation group, 2 mL physiological saline containing 5×10<'6> BrdU-labeled EPCs was injected into the coronary artery, while in the model control group, simple physiological saline of the same amount was given. At 1,4, and 8 weeks after transplantation, dogs were sacrificed for harvesting myocardial tissue.MAIN OUTCOME MEASURES: Myocardial infarction was confirmed by hematoxylin-eosin staining. Myocardial angiogenesis was observed by BrdU immunohistochemical staining. The number of infarcted myocardial vessels was calculated by yon Willebrand (vW) factor staining.RESULTS: There was plenty of scar tissue, flbroblasts, and small vessels in the myocardial infarction region. In the cell transplantation group, brown yellow particles (BrdU-positive expression) appeared in some nuclei in small vessels from infarcted myocardium. Newly formed vessels were not found in the model control group. In the cell transplantation group, brown yellow particles (vW factor-positive expression) appeared in the cytoplasm of the vascular endothelial cells in the myocardial ischemia and infarction regions, vW factors were not expressed in the model control group. At 1, 4, and 8 weeks after myocardial infarction,there was no significant difference in vessel counts no matter in myocardial ischemia region or in myocardial infarction region between the call transplantation and model control groups.CONCLUSION: EPCs derived from UCB of pregnant dog can participate in the formation of blood vessels but can not promote angiogenesis after acute myocardial infarction.

Background Induced pluripotent stem cells (iPSCs)can differentiate into various types of somatic cells without causing ethical controversy and immune rejection in clinical activity,which is similar to differentiation ability of embryonic stem cells.So,iPSCs may be used as seed cells for tissue engineering corneal endothelial reconstruction.Objective The present study was to survey the morphologic change of iPSCs after coculture with corneal endothelium cells(CECs) under the atomic force microscopy(AFM).Methods Rabbit CECs and human MMC-iPSCs were isolated and cultured respectively.The iPSCs were identified with the marker by immunochemistry.iPSCs passaged for 7 days were then cultured with 60％ confluent CECs to establish the co-culture model.The surface morphology and cellular membrane ultrastructure of differentiated iPSCs after induced by CECs were examined by AFM combination with inverted microscope,and compared with CECs and undifferentiated iPSCs.Results Thelengthand width were(66.93±10.48)μm and (44.85 ± 8.14) μm in CECs,(12.51±1.40)μm and (10.93 ±1.69) μm in uninduced iPSCs,and(36.12±10.29) μm and(31.53±9.65)μm in CECs-induced iPSCs.Both the length and width values of CECs-induced iPSCs were statistically bigger than those uninduced iPSCs,with significant differences between them (P＜0.05),but no significant difference was seen in the width valne of CECs-induced iPSCs in comparison with CECs(P＞0.05).The convex structure of CECs cytomembrane surface showed the digitation in shape with the size and height(2.11 ± 1.03) μm and (115.68±92.08) nm respectively,and the concave structure of cytomembrane surface of CECs was fenestrae-like depression and the size was (1.49 ± 0.65) μm.The numerical valuc of mean square root roughness (Rq)and average roughness (Ra)of cytomembrane surface of CECs were(39.20±7.82)nm and (30.37±5.32)nm respectively.The convex surface of cytomembrane of iPSCs was granular-like in shape with size and height(0.39±0.22)μm and(13.11±9.18)nm respectively.The concave surface of cytomembrane of iPSCs was worm-eaten-like concave with the size(0.34±0.18)μm.The numerical value of Rq and Ra of geometrical parameters of cytomembrane surface of iPSCs were (26.60 ± 4.93)nm and (9.97 ± 3.78) nm respectively.The convex surface of cytomembrane of induced iPSCs was digital-like in shape with the size and height (1.91±0.76) μm and(106.55±77.27) nm respectively.The concave surface of cytomembrane of induced iPSCs was fenestrae-like depression and the size of concave was(1.6l±1.25) μm.The numerical value of Rq and Ra on surface of cytomembrane of induced iPSCs was (57.33± 12.80) nm and (43.63± 11.17) nm respectively.The numerical values of the size and height of convex,the size of concave,Rq and Ra on surface of cytomembrane in induced iPSCs were statistically bigger than in iPSCs(P＜0.05)and were not significant differences in comparison with CECs (P＞0.05).Conclusions Morphology of iPSCs translate toward the CECs after induce for 7 days under the AFM.This outcome lays the foundation for further study on iPSCs.

Animals ; COS Cells ; Female ; Hepatitis B Core Antigens ; genetics ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B Vaccines ; immunology ; Interleukin-12 ; genetics ; Mice ; Mice, Inbred BALB C ; Vaccines, DNA ; immunology