OBJECTIVETo investigate the protective effect of phosphodiesterase type 5 inhibitors (tadalafil) on the testis following testicular ischemia-reperfusion injury in rats.

METHODSEighty-four healthy adult male SD rats were randomly and equally divided into groups A (sham operation), B (testicular torsion + low-dose tadalafil), C (testicular torsion + high-dose tadalafil), and D (testicular torsion + placebo). Models were established in the latter three groups by 7200 torsion of the right testis for 2 hours. The animals in groups A and B were treated by gavage with tadalafil at the dose of 0. 5 mg per kg per day, those in group C at 2 mg per kg per day, and those in group D with saline at the same dose. After 3, 7, and 14 days of treatment, the torsioned testes were harvested for evaluation of the superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the testis tissue. The pathological changes in the testis were observed under the light microscope.

RESULTSAt 3, 7, and 14 days, the SOD activity was (254.46 +/- 7.43), (278.49 +/- 8.33), and (317.99 +/- 3.31) nU/mg prot in group B, and (277.12 +/- 8.80), (309.40 +/- 2.14), and (320.39 +/- 4.72) nU/mg prot in group C, all obviously higher than in D ([223.21 +/- 4.65], [231.45 +/- 4.16] and [248.28 +/- 5.74] nU/mg prot), while the MDA content was lower in the former two groups than in the latter. At 3 and 7 days, the SOD activity was significantly higher and the MDA level significantly lower in group C than in B (both P < 0.01) , while at 14 days, neither showed any remarkable differences between the two groups (P > 0.05). No obvious histopathological change was observed in the testis tissue of group A. At 3 and 7 days, pathological examination of the testis tissue revealed significant differences in the number of seminiferous epithelial layers, testicular histological score, and seminiferous tubule diameter in group B (P < 0.01), but the three indexes at 14 days in group B and at 7 days in group C exhibited no remarkable differences from those at 14 days in group A.

CONCLUSIONTadalafil can alleviate testicular ischemia-reperfusion injury following testis torsion/detorsion in a time- and dose-dependent manner.

Animals ; Biomarkers ; metabolism ; Carbolines ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Male ; Malondialdehyde ; metabolism ; Phosphodiesterase 5 Inhibitors ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Seminiferous Tubules ; pathology ; Spermatic Cord Torsion ; complications ; Superoxide Dismutase ; metabolism ; Tadalafil ; Testis ; blood supply ; metabolism ; pathology ; Time Factors

Objective To investigate the proportion and function of CD_4~+ CD_(25)~+ regulatory T cells (CD_4~+ CD_(25)~+ Tr)in unexplained recurrent spontaneous abortion(URSA).Methods(1)Proportion measurement:the proportion of CD_4~+ CD_(25)~+ Tr cells in peripheral blood was measured by double-label flow cytometric analysis.The samples were taken from 15 URSA women,15 normal non-pregnancy women and 13 normal pregnancy women.(2)Function measurement:CD_4~+ CD_(25)~+ Tr ceils and CD_4~+ CD_(25)~+ T ce]ls were extracted from peripheral blood lymphocytes by the microbeads separation.The purity of CD_4~+ CD_(25)~+ Tr cells and CD_4~+ CD_(25)~+ T cells was measured by flow cytometry.The growth inhibitory effect of CD_4~+ CD_(25)~+ Tr cells on CD_4~+ CD_(25)~+ T cells was assessed in vitro.Results The proportion of CD_4~+ CD_(25)~+ Tr cells was decreased significantly in URSA women(6.9?1.8)% than that in normal non-pregnancy women[(10.8?1.1)%] (P0.05).Conclusion The results suggest that decrease in proportion and function of CD_4~+ CD_(25)~+ Tr cells may be associated with URSA.

BACKGROUNDPatients with severe full-thickness burn injury suffer from their inability to maintain body temperature through perspiration because the complete destructed sweat glands can not be regenerated. Bone marrow-derived mesenchymal stem cells (BM-MSCs) represent an ideal stem-cell source for cell therapy because of their easy purification and multipotency. In this study, we attempted to induce human BM-MSCs to differentiate into sweat gland cells for sweat gland regeneration through ectodysplasin (EDA) gene transfection.

METHODSThe dynamic expression of EDA and EDA receptor (EDAR) were firstly observed in the sweat gland formation during embryological development. After transfection with EDA expression vector, human BM-MSCs were transplanted into the injured areas of burn animal models. The regeneration of sweat glands was identified by perspiration test and immunohistochemical analysis.

RESULTSEndogenous expression of EDA and EDAR correlated with sweat gland development in human fetal skin. After EDA transfection, BM-MSC acquired a sweat-gland-cell phenotype, evidenced by their expression of sweat gland markers by flow cytometry analysis. Immunohistochemical staining revealed a markedly contribution of EDA-transfected BM-MSCs to the regeneration of sweat glands in the scalded paws. Positive rate for perspiration test for the paws treated with EDA-transfected BM-MSCs was significantly higher than those treated with BM-MSCs or EDA expression vector (P < 0.05).

CONCLUSIONSOur results confirmed the important role of EDA in the development of sweat gland. BM-MSCs transfected with EDA significantly improved the sweat-gland regeneration. This study suggests the potential application of EDA-modified MSCs for the repair and regeneration of injured skin and its appendages.

Adult ; Animals ; Blotting, Western ; Bone Marrow Cells ; cytology ; Cell Proliferation ; Cells, Cultured ; Ectodysplasins ; genetics ; metabolism ; Female ; Flow Cytometry ; Humans ; Immunohistochemistry ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Pregnancy ; Receptors, Ectodysplasin ; Reverse Transcriptase Polymerase Chain Reaction ; Sweat Glands ; cytology ; metabolism ; Transfection ; Young Adult

OBJECTIVETo investigate the influence of biological behavior of epithelial cells on the hair follicles and sebaceous glands (HFSG) structure in keloids (K).

METHODSThe expression of intercellular adhesion molecule (ICAM)-1, D-related human leucocyte antigen (HLA-DR), and cytokeratin (CK) 14 on epithelial cells and the amount of activity T-lymphocytes were detected in specimens of keloid edge and normal skin with immunohistochemical and histological methods.

RESULTSIn comparison with normal skin specimens, epithelial cells were proliferated in K-HFSG presented structural aberration and disintegrate or abnormally to form solid-epithelial island-like structure, and the density of HSFG with hyperplasia and the ageing scar in keloids was apparently decreased. They strongly expressed ICAM-1, HLA-DR, and CK14 in the epithelial cells, there were many immunologic cells which expressed CD4, CD45RO, and interferon (IFN)-gamma around the K-HFSG. The expressed level of epithelial cells was positively correlated with the density of immunologic cells nearby K-HFSG.

CONCLUSIONIt could be concluded that the reactivity with hyperplasia and immunoinduction of epithelial cells might be associated with the destruction of the some HFSG structure in the keloids.

Adolescent ; Adult ; Cell Proliferation ; Child ; Child, Preschool ; Epithelial Cells ; pathology ; Female ; HLA-DR Antigens ; metabolism ; Hair Follicle ; pathology ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Keloid ; immunology ; pathology ; Keratin-14 ; metabolism ; Male ; Middle Aged ; Sebaceous Glands ; pathology ; Young Adult

This study was aimed to construct a two-dimensional culture system by using osteoblasts induced from human marrow mesenchymal stem cells (MSC) and to investigate its support effect on survival of hematopoietic stem/progenitor cells for umbilical cord blood (UCB) ex vivo. MSCs were isolated from adult human bone marrow and were cultured, the second generation of MSCs were induced into osteoblasts which were irradiated with 20 Gy gamma rays in a Cobalt 60 source and confluenced into a feeder layer. CD34(+) cells were selected from fresh umbilical cord blood samples by using Microbead Kit of MiniMACS and seeded into the two-dimensional culture system to culture ex vivo without exogenous cytokines. By using colony-forming assay, high proliferative potential colony-forming cell assay, and long-term culture initiating cell assay, the ability of the two-dimensional system to culture HSCs/HPCs was observed. The results showed that the osteoblasts induced from bone marrow MSC in constructed two-dimensional culture system displayed more significant support effect on survival of hematopoietic stem/progenitor cells from umbilical cord blood (UCB) ex vivo, compared with other culture systems, especially on long term HSCs survival ex vivo. It is concluded that the two-dimensional culture system constituted by osteoblasts induced from human MSCs has certain ability of supporting maintenance and multipotency of HSCs/HPCs from umbilical cord blood in vitro, especially sustaining survival of HSC in long-term culture. It has also been proved that osteoblasts play a crucial role in regulation of HSC growth.
Bone Marrow Cells ; cytology ; Cell Differentiation ; Coculture Techniques ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology ; physiology

OBJECTIVETo investigate the migrating effect of adipose derived stem cells (ADSCs) on the wound model of human epidermal keratinocyte (HEKa).

METHODSRat ADSCs (rADSCs) were isolated and cultured (n = 10), rADSCs were direct co-cultured with HEKa cells in experiment group (experimental group, n = 10). In the control groups, rADSCs were indirect co-cultured with HEKa cells in transwell chamber (indirect group, n = 8), or HEKa was cultured alone (single group, n = 8). Then confluent HEKa cells were scraped to establish a wound model under invert microscope. After scraped 24, 48, and 72 h, cell numbers of which migrated across the edge of the wound was measured, the rate of wound healing was calculated by using SigmaScan Pro 5 software, and the proliferating effect of rADSCs on HEKa were examined by incorporation of [(3)H] thymidine.

RESULTSThe cells migrated across the edge of wound after 24 hours in experimental group, indirect group, and single group were (9.2 + or - 0.2), (5.0 + or - 0.3), (4.2 + or - 0.3), and were (58.5 + or - 0.4), (26.5 + or - 0.3), (20.7 + or - 0.5) 48 hours after, and were (125.8 + or - 0.4), (43.0 + or - 0.5), (35.6 + or - 0.5) cells/HP 72 hours after, respectively; the numbers were all significantly higher in experimental group than those in control groups (P < 0.05). The rates of wound healing after scraped 72 hours were 61.0% + or - 3.0%, 35.0% + or - 2.5% and 32.0 + or - 2.1%, the outcome in experimental group was significantly better than in the control groups (P < 0.05). And the thymidine feeding displayed the proliferation of HEKa in the three groups were (1440 + or - 210), (1050 + or - 280) and (1130 + or - 390) cpm/10(5) cell, and there was significant difference between the experimental and the control groups (P < 0.05).

CONCLUSIONSThe rADSCs can promote the migration of HEKa by direct contact with it.

Adipose Tissue ; cytology ; Animals ; Cell Count ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Epidermis ; cytology ; Humans ; Keratinocytes ; cytology ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; Wound Healing

OBJECTIVETo observe the expression characteristics of EGFR and FGFR-2 in normal skin from fetal and adult, and attempted to probe the molecule mechanism of fetal scarless healing.

METHODSThe skin samples of fetal and adult were taken from abortive fetus of obstetrics unit and donor site of plastic operation patients in our burn unit, respectively. EGFR and FGFR-2 were used as the biochemical markers for reparative cells. Immunohistochemistry staining technique was employed to determine the expressive levels of different epithelial cells markers.

RESULTSThere were EGFR and FGFR-2 antibody positive cells in normal skin from fetal and adult, but the expressive levels of EGFR and FGFR-2 protein had apparent difference, with the time of fetation increasing, the EGFR and FGFR-2 positive expression rate became stronger gradually. The number of FGFR-2 antibody positive cells found in adult skin was much more than that in fetal skin.

CONCLUSIONThere were the inherent differences of EGFR and FGFR-2 antibody immunohistochemistry staining in cells of adult and fetal skin. which may be an essential facet of fetal scarless healing.

Adult ; Epithelial Cells ; metabolism ; ErbB Receptors ; metabolism ; Female ; Fetus ; metabolism ; Humans ; Immunohistochemistry ; Receptor, ErbB-2 ; metabolism ; Skin ; metabolism ; Wound Healing

OBJECTIVETo evaluate the effect of absolute alcohol treatment for renal cyst with percutaneous puncture and catheterization.

METHODSThis report presented 64 cases of renal cysts, 34 cases were treated with percutaneous puncture (A group) and 30 cases with percutaneous catheterization (B group). According to the size, the cysts were divided into 2 groups, more than 6 cm in diameter and less than 6 cm in diameter.

RESULTSAll the patients were followed up for 3 - 12 months by CT or B ultrasonography. Striking difference of the therapeutic results were existed when cystS were more than 6 cm in diameter.

CONCLUSIONPercutaneous catheterization is applicable to the sclerosing treatment of renal cyst whose diameter is more than 6 cm.

Adult ; Aged ; Catheters, Indwelling ; Combined Modality Therapy ; Ethanol ; administration & dosage ; Female ; Humans ; Injections, Intralesional ; Kidney Diseases, Cystic ; diagnostic imaging ; therapy ; Male ; Middle Aged ; Paracentesis ; methods ; Sclerosing Solutions ; administration & dosage ; Treatment Outcome ; Ultrasonography, Interventional

The promoter of NAR gene in Magnaporthe grisea was isolated and sequenced. The promoter sequences contained the "TATA" box, the "CAAT" box, and binding sites for fungal regulatory proteins. Programs that predict promoter sequences indicated that promoter sequence lies between locations 430 and 857 of the NAR promoter fragment. GFP expression under the NAR promoter and NAR transcript analysis revealed that this promoter is activated primarily at the mycelial stage in the rice blast fungus and could be used to express native or extrinsic genes in the mycelia of the rice blast fungus.
Base Sequence ; Cloning, Molecular ; Fungal Proteins ; genetics ; Gene Expression ; genetics ; Hyphae ; genetics ; Magnaporthe ; genetics ; Molecular Sequence Data ; Promoter Regions, Genetic ; genetics ; Transcriptional Activation ; genetics

GATA-3 plays a central role in the Th2-mediated immunoreaction. This study was aimed to construct and select plasmid vectors of siRNA which can effectively and specifically suppress the gene expression of GATA-3. Plasmid including PSi338, PSi717 and PSi1232 were designed and constructed for GATA-3 regarded as target gene and transfected into murine P815 cells. Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot were used to detect the inhibition of GATA-3 mRNA as well as its protein in P815 cells. The results demonstrated that the expressions of mRNA and protein of GATA-3 in P815 cells were inhibited significantly by both of PSi338 and PSi717. It is concluded that PSi338 and PSi717 siRNA plasmid vectors have been successfully constructed, and both vectors are effective and specific siRNA plasmids for suppressing GATA-3 gene expression.
Animals ; Down-Regulation ; GATA3 Transcription Factor ; metabolism ; Genetic Vectors ; Mice ; Plasmids ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Th2 Cells ; immunology ; Transfection