Child, Preschool ; Cleft Lip ; complications ; surgery ; Cleft Palate ; complications ; surgery ; Hernia, Diaphragmatic ; complications ; surgery ; Hernias, Diaphragmatic, Congenital ; Humans ; Male

The technique conditions of decolonization of fermentation broth were successively optimized using single factor assay and orthogonal layout method in Cordyceps jiangxiensis. The optimal condition of decolorization was investigated to be 3g/100mL active carbon, 5 min absorption time, pH5 of fermented broth and 25℃absorption temperature. Under the optimal condition, the maximum decolorization rate of fermented broth reached 89. 6% , simultaneously 10. 7% consuming rate of exopolysaccahride was minimum. Subsequently, the extract condition of exopolysaccharide of C. jiangxiensis was further optimized by orthogonal layout design. The maximum exopolysaccharide production was 0. 38 g/L under the optimal condition, i. e. firstly fermented filtrate decolorized and deproteined was concentrated to 1/7 of its total volume, secondly concentration broth was mixed with four times its volume of absolute ethanol and stirred vigorously, lastly precipitation of exopolysaccharide proceeded at 4℃for 16 hrs and the exopolysaccharide collected by centrifugal ion and dryness.

To explore the plasticity of human amniotic mesenchymal cells(hAMCs) into cardiomyocyte-like cells,hAMCs were isolated from human amnion with collagenase digestion.Phenotype of the isolated cells was analyzed by flow cytometry(FCM).hAMCs were treated with 5-azacytidine and basic fibroblast growth factor(bFGF) to investigate their ability of differentiation into cardiomyocytes.The induced differentiated cells were evaluated by immunofluorescence for desmin and ?-actin expression and by RT-PCR for Nkx2.5,GATA-4 and alpha-myosin heavy chain(?-MHC) mRNA expression.The results showed that,after primary culture,hAMCs could reach a confluence of 80% with swirl like growth at 6 days.The cells proliferated rapidly after passages with a 100% confluence at 3~4 d.hAMCs were positive expression of CD44 and vimentin,but negative for CK19.After induced differentiation at 8~10d,the differentiated cells have close-up arranged with long spindle-shape.At 2 weeks and 4 weeks,induced cells expressed ?-actinin and cardiac-specific transcription factor Nkx2.5.Expressions of GATA-4 and desmin can be detected but ?-MHC can not in the hAMCs both before and after the induction.In conclusion,hAMCs have the ability of differentiation into cardiomyocyte-like cells,which means that hAMCs can be regarded as candidate cells for cellular cardiomyoplasty(CCM).

OBJECTIVETo observe the therapeutic effect of electroacupuncture (EA) at Quchi (LI 11) on blood pressure and blood plasma catecholamines in the patient of essential hypertension and to investigate the mechanism.

METHODSSixty cases of essential hypertension were randomly divided into an EA group (n=30) and a control group (n=30). In the EA group, bilateral Quchi (LI 11) were selected; and in the control group, western medicine Nicardipine was taken. The variation of blood pressure and blood plasma catecholamines were examined before and after the treatment.

RESULTS(1) After treatment, there were significant reduction in the levels of systolic blood pressure and diastole blood pressure in both groups (P < 0.01); (2) After treatment, significant reduction in levels of adrenaline and noradrenaline were also found in both groups (P < 0.01), however, no significant differences in the level of dopamine were observed in both groups (P > 0.05); (3) The effective rate of 66.7% in the EA group was similar to that of 70.0% in the control group (P > 0.05).

CONCLUSIONBoth EA at Quchi (LI 11) and western medicine are able to beneficially regulate blood pressure of patients with essential hypertension through adjusting blood plasma catecholamines.

Acupuncture Points ; Aged ; Blood Pressure ; Catecholamines ; blood ; Electroacupuncture ; Female ; Humans ; Hypertension ; blood ; physiopathology ; therapy ; Male ; Middle Aged

OBJECTIVETo investigate the expression of programmed cell death 5 (PDCD5) in tissues of normal human prostate (NP), benign prostatic hyperplasia (BPH), and prostate cancer (PCa) in order to assess the clinical role of PDCD5 in PCa.

METHODSPDCD5 expression was determined by EnVision immunohistochemical staining in formalin-fixed and paraffin-embedded specimens obtained from 12 subjects with NP, 22 with BPH, and 22 with PCa. In addition, PCa cases were classified as low/middle-risk (Gleason sum < or = 7) and high-risk (Gleason sum >7) on the basis of Gleason grade. Positive expression rates and intensity of PDCD5 protein were observed under light microscope and analyzed with computer imaging technique. Expression of PDCD5 was compared among different prostatic tissues.

RESULTSThe expression of PDCD5 was significantly lower in tissue of PCa than in tissues of NP and BPH (P<0.01). However, there was no significant difference in PDCD5 expression between tissues of NP and BPH. In addition, the expression of PDCD5 was further downregulated with the increase of Gleason sum in PCa.

CONCLUSIONSBy downregulating apoptosis, low PDCD5 expression may play an important role in the occurrence and development of PCa. PDCD5 is supposed to have a potential clinical value to be a new predictor of progression and target of gene therapy in PCa.

Aged ; Apoptosis ; Apoptosis Regulatory Proteins ; analysis ; physiology ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Proteins ; analysis ; physiology ; Prostate ; chemistry ; Prostatic Neoplasms ; chemistry ; etiology ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; analysis

Objective To investigate the effects of the enhancer of zeste homolog 2 (EZH2) gene on cell growth and invasion of the nasopharyngeal carcinoma (NPC).Methods Recombinant lentivirus vector for shRNA delivery of EZH2 was constructed and transfected into 293FT cells. After collecting the viral particles, the NPC cell line 5-8F cells were transfected. The effects of EZH2 silence on cell proliferation and cell cycle were detected using MTT assay,plate colony formation assay and flow cytometry.The migration and invasion of 5-8F cells were determined by wound healing assay and matrigel invasion assay,respectively.The expressions of EZH2 and epithelial-mesenchymal transition (EMT) -related markers at mRNA and protein levels were examined by real-time PCR and Western blot respectively.Results The expressions of EZH2 mRNA and protein in the transfected 5-8F cells were obviously reduced. MTT assay showed that EZH2 downregulation significantly inhibited the growth of 5-8F/shEZH2 cells( P ＜ 0.001 ).Colony formation rate (84.44％ ) of 5-8F/shEZH2 cells was lower than control (31.56％,P =0.001 ).Cell cycle analysis showed that most 5-8F/shEZH2 cells were arrested in G0/G1 phase,with a very low ratio of cells in S phase.Wound healing assay indicated that the migration ability of cells silencing EZH2 decreased significantly,and the 48-hour relative migration distance of 5-SF/ShEZH2 cells and control cells was 0.58 ± 0.05,and 0.81 ± 0.02,respecptively(P ＜ 0.000).Matrigel invasion assay,showed the invasive capacity of cells silencing EZH2 was significantly inhibited,with less penetrating cells (72.23 ± 4.08 ) compared to control( 150.95 ± 16.27),P ＜ 0.000.The mRNA expressions of epithelial markers E-cadherin and Keratin 18 in the cells silencing EZH2 increased by 177％ and 158％ respectively,and the mRNA expressions of mesenchymal markers β-catenin and N-cadherin decreased by 18.04％ and 41.18％ respectively. Similar results also were obtained with Western blot analysis. Conclusion EZH2 significantly enhanced the proliferation and invasion of nasopharyngeal carcinoma cells in vitro,which might be mediated by inducing EMT.

The aim of this study was to evaluate whether human placenta CD133(+) cells have an ability to reconstitute long-term hematopoiesis. Magnetic-activated cell sorting (MACS) was applied to enrich human placental CD133(+) cells. The isolated human placental CD133(+)cells of four different densities were established by limiting-dilution assay and primary fetal bone marrow stromal cells separated from bone marrow as feeder layer cells were co-cultured in long-term culture system so as to observe the incidence of long-term culture initiating-cells (LTC-IC) and their ability of proliferation and differentiation.The results showed that human placenta derived CD133(+) cells contained LTC-IC with frequency of 1/645 which have an ability to proliferate and differentiate into granulocyte/macrophage colony-forming units (CFU-GM) and mixed colony-forming units (CFU-Mix). In all LTC-IC positive wells, 71.43% form only CFU-GM and 28.57% display both CFU-GM and CFU-Mix formation. It is concluded that human placental CD133(+) cells possess LTC-IC with colony-forming capacity of hematopoietic early progenitor cells.
AC133 Antigen ; Antigens, CD ; immunology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Separation ; Colony-Forming Units Assay ; Female ; Glycoproteins ; immunology ; Hematopoietic Stem Cells ; cytology ; Humans ; Peptides ; immunology ; Placenta ; cytology ; immunology ; Pregnancy

OBJECTIVETo study the role of programmed cell death 5 (PDCD5) protein in the oncogenesis and development of renal clear cell carcinoma and its association with the prognosis of the malignancy.

METHODSPDCD5 expression was assayed immunohistochemically in 46 cases of human renal clear cell carcinoma, and the patients' survival was followed up.

RESULTSPDCD5 staining in the adjacent normal tissue of the tumor was significantly stronger than that in the tumor tissue, and PDCD5 expression was significantly correlated with the tumor grade, stage and prognosis. The tumors of high grade with strong invasive ability had much less PDCD5 expression and lighter staining. The three- or five-year survival rates of patients positive for PDCD5 expression was much higher than that of patients negative for PDCD5 expression.

CONCLUSIONPDCD5 is a potent inhibitor of malignant transformation of renal clear cell carcinoma and may serve as a major predictor for evaluating the malignant potential and prognosis of the tumor.

Adult ; Aged ; Apoptosis Regulatory Proteins ; biosynthesis ; Carcinoma, Renal Cell ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Kidney Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; biosynthesis ; Prognosis ; Retrospective Studies ; Survival Analysis

OBJECTIVETo study the effect of SREBP-1c silencing on lipid metabolism and expression of inflammatory chemokines in a NAFLD model with endoplasmic reticulum stress.

METHODNAFLD model was established in L02 cells treated with oleic acid. SREBP-1c expression was inhibited using RNA interference with a p Silencer-1.0-U6-4476 vector. After transfection with p Silencer-1.0-U6-4476 or control vector for 0 h, 24 h, 48 h and 72 h, the extent of fatty degeneration was shown by Oil Red O staining. The mRNA and protein expression of inflammatory chemokine CCL2 and basic fibroblast growth factor-21 (FGF21) were determined by real time PCR and Western blot respectively.

RESULTSSREBP-1c silenced L02 cells showed fat droplets with smaller diameter and attenuated fatty deposition, as compared with control cells. The relative CCL2 mRNA levels in SREBP-1c silencing vector transfected L02 cells were 1.03+/-0.11 for 0 h, 1.11+/-0.21 for 24 h, 0.88+/-0.16 for 48 h, and 1.05+/-0.15 for 72 h, which showed no significant difference as compared with control cells (P>0.05, respectively). In addition, no difference was found between the different time points within the same group (P>0.05). However, CCL2 protein levels in SREBP-1c silenced cells were 1.19+/-0.15, 1.07+/-0.18, 0.48+/-0.14, and 0.05+/-0.24 after transfection for 0 h, 24 h, 48 h, and 72 h respectively, which were significantly downregulated as compared to the control group (P<0.01). And CCL2 protein levels between different time points in SREBP-1c silenced cells were also distinct (P<0.01). The relative FGF21 mRNA levels in SREBP-1c silenced L-02 cells were 1.01+/-0.08, 0.91+/-0.22, 0.98+/-0.20, and 1.02+/-0.12 for 0 h, 24 h, 48 h, and 72 h respectively, which were not statistically different as compared with the corresponding control cells. Statistic difference of FGF21 mRNA levels in SREBP-1c knockdown cells of different time points was not found (P>0.05). In striking contrast, robust down regulation of FGF21 protein in SREBP-1c silenced cells was observed, with 0.81+/-0.05, 0.66+/-0.12, 0.58+/-0.08 and 0.19+/-0.13 after transfection for 0 h, 24 h, 48 h and 72 h respectively, as compared to control group (P<0.01). And differences in FGF21 protein level between different time points in SREBP-1c silenced cells were also demonstrated (P<0.01).

CONCLUSIONSREBP-1c knockdown attenuated fatty deposition in oleic acid treated L02 cells. In addition, silencing of SREBP-1c expression reduced expressions of CCL2 and FGF21 proteins posttranscriptionally, which may play a role in endoplasmic reticulum stress induced inflammatory response in NAFLD.

Cell Line ; Chemokine CCL2 ; metabolism ; Endoplasmic Reticulum Stress ; Fibroblast Growth Factors ; metabolism ; Gene Knockdown Techniques ; Hepatocytes ; metabolism ; Humans ; Lipid Metabolism ; RNA Interference ; Sterol Regulatory Element Binding Protein 1 ; genetics

OBJECTIVETo compare the change of lung tissue and vasopermeability between the vascular endothelial cells special cdc42-deficient heterozygous mice and the non-knockout mice in acute lung injury.

METHODSThe mice with vascular endothelial cell-specific expression of cre recombinase were crossed with cdc42(flox/flox) mice. The cdc42(flox/+)Cre(+/-) F1 offspring mice were crossed back with cdc42(flox/flox) mice, resulting in the F2 generation mice with three genotypes, namely cdc42(flox/+)Cre(+/-), cdc42(flox/flox)Cre(-/-) and cdc42(flox/+)Cre(+/-). The heterozygous mice with cdc42(flox/+)Cre(+/-) genotype were selected as the model mice, with the other two genotype groups as the control. After intratracheal instillation of 2 mg/kg LPS to induce acute lung injury, the mice were sacrificed to examine the lung pathologies, lung wet/dry ratio and lung microvascular permeability.

RESULTSThe heterozygous mice with cdc42 gene knockout (cdc42(flox/+)Cre(+/-)) showed no significant differences from the two control groups in the lung pathological score, lung wet/dry ratio or the lung microvascular permeability coefficient.

CONCLUSIONThere were no significant difference on lung tissue and vasopermeability between the vascular endothelial cells special cdc42-deficient heterozygous mice and the non-knockout mice.

Acute Lung Injury ; pathology ; Animals ; Capillary Permeability ; Endothelial Cells ; pathology ; Integrases ; genetics ; Lung ; blood supply ; pathology ; Mice ; Mice, Knockout ; cdc42 GTP-Binding Protein ; genetics