Objective To explore the high risk factors, pathogenesis and methods of early diagnosis of periventricular leukomalacia(PVL) in premature infants.Methods The history of intrauterine hypoxia-ischemia was investigated in premature infants;TORCH-IgM antibodies in cord blood of premature infants were measured by ELISA; Nitric oxide (NO) levels in their cord blood were determined by nitric acid reducing enzyme means. Results Thirty-nine of 52 premature infants in PVL group had a history of intrauterine hypoxia-ischemia; TORCH-IgM antibody positive rate in cord blood of premature infants in PVL group was significantly higher than that in control group(P

OBJECTIVETo obtain the monoclonal antibody against hexon protein of human adenovirus.

METHODSBALB/c mice were immunized with purified recombinant hexon protein, and the spleen cells of the mice were isolated and fused with myloma cells. Four hybridoma cell strains were screened by indirect ELISA and cultured, and the sensitivity, specificity and virus neutralizing activity were analyzed with ELISA, Western blotting and neutralizing test.

RESULTSThe mouse ascites produced by these hybridoma cells contained specific monoclonal antibodies against hexon protein of human adenovirus as identified by ELISA and Western blot, and the antibody generated by 4C6 strain showed human adenovirus type 3-neutralizing activity.

CONCLUSIONThe monoclonal antibodies against hexon protein with high specificity have been successfully obtained, and these antibodies can be useful in developing assays for early diagnosis of HAdV3 infection and also in study of therapeutic drugs of the infection.

Adenoviruses, Human ; chemistry ; immunology ; Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibodies, Viral ; biosynthesis ; immunology ; Blotting, Western ; Capsid Proteins ; biosynthesis ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; immunology

OBJECTIVETo investigate whether calcineurin (CaN) contribute to tumor necrosis factor alpha (TNF-alpha)-induced cardiomyocyte hypertrophy.

METHODSThe protein content was assayed with lowry's method. The cardiomyocytes volumes were measured by computer photograph analysis system. The protein synthesis was assayed with [3H]-leucine incorporation method. [Ca2+]i transient was measured by Till image system by cell-loading Fura-2/AM. The expression of CaN was determined by Western blot.

RESULTS(1) (CsA (0.2 micromol/L), a selective CaN inhibitor, significantly suppressed the increase of protein content, [3H]-leucine incorporation and cell size induced by TNF-alpha. (2) CsA (0.2 micromol/L) significantly suppressed the elevation of the amplitude of the spontaneous Ca2+ transients induced by TNF-alpha in cultured ventricular myocytes from the neonatal rat. (3) TNF-alpha significantly increased the expression of CaN.

CONCLUSIONCa(2+) -CaN signaling pathway are involved in cardiomyocyte hypertrophy induced by TNF-alpha in rats.

Animals ; Calcineurin ; metabolism ; Calcium Signaling ; Cardiomyopathy, Dilated ; metabolism ; pathology ; Cells, Cultured ; Female ; Male ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; pharmacology

Objective　To explore the effects of hypoxia on the syntheses and secretion of adrenomedullin (AM), calcitonin gene related peptide (CGRP) and c-type natriuretic peptide (CNP) and the relationship between these peptides. Methods　Rat models were established with hypoxia for 10, 20 and 30 d respectively and rats under normal altitude were served as control. Pulmonary artery pressure and the maximum increasing speed of right ventricle (RVdp/dtmax) were measured in every group. The dynamic changes of AM, CGRP and CNP concentrations in plasma were studied with radioimmunoassay. Results　During hypoxia, pulmonary artery pressure and RVdp/dtmax were enhanced. Plasma AM and CNP concentrations were increased while CGRP was decreased significantly. The plasma level of AM had positive correlation with that of CNP, but negatively correlated with that of CGRP. Conclusion　Results indicate that hypoxia may cause pulmonary artery pressure change and right ventricle has compensatory reaction to hypoxic pulmonary hypertension. Dynamic changes of plasma AM, CGRP and CNP concentrations can be regarded as indexes for condition of illness.

OBJECTIVETo study the different features of hyperplasia in castrated and uncastrated mice after testosterone (T) treatment.

METHODSForty-eight BALB/c mice were randomly divided into 6 groups of 8 in each: castrated (A), uncastrated (B) , castrated + low T (C), uncastrated + low T (D), castrated + high T (E), uncastrated + high T (F). Groups C and D were treated with testosterone solution at the dose of 12.5 mg/(kg d) and Groups E and F at 125 mg/(kg d) for 20 consecutive days, while Groups A and B received saline only. All the mice were sacrificed on the 21st day, their ventral and dorsal prostate glands weighed and their pathological features studied.

RESULTSAtrophic prostates were observed in Group A, but normal in Group B; prostatic hyperplasia was found in both Group C and D, but more obvious in the latter (P <0.05); and a slightly higher degree of hyperplasia was noted in Groups E and F than in C and D. There was an increase in serum T and vascular endothelial growth factor (VEGF) concentration and a decrease in serum estrogen (E2) concentration in the testosterone treated groups.

CONCLUSIONBoth castrated and uncastrated mice develop prostate hyperplasia after short-term testosterone treatment, although in different degrees and with different features, which may help further the studies on the association of castration and androgen with prostate diseases.

Animals ; Hyperplasia ; Male ; Mice ; Mice, Inbred BALB C ; Orchiectomy ; Prostate ; pathology ; Prostatic Hyperplasia ; drug therapy ; pathology ; Testosterone ; therapeutic use

OBJECTIVETo construct vector pEGFP-C1-hTERT-ribozyme (pGTRz-U6) and its mutant (pGTmRz-U6) against hTERT containing U6 promoter, then transfect them into human liver cancer cell line SMMC7721 to observe the action of the human telomerase catalytic subunit (hTERT) hammerhead ribozyme on proliferation and apoptosis of human liver cancer cell SMMC7721.

METHODSEukaryotic expressing vector pGTRz-U6 and mutant pGTmRz-U6 were constructed and transfected into SMMC7721 using Lipofectamine2000 Reagent, with pEGFP-C1 as the control group. After strict screening by G418, positive clones were cultured; the amount of expression of ribozyme and hTERT was detected by RT-PCR; cell proliferation by MTT; telomerase activity by TRAP and silver staining assay; cell apoptosis by FCM.

RESULTSWe found that the two ribozymes were expressed persistently in SMMC7721; different expression levels (P < 0.01) of hTERT among SMMC7721-Rz, SMMC7721-mRz and SMMC7721-pEGFP-C1 was exhibited by the analysis of variance with SPSS software. The difference between SMMC7721-Rz and the others is significant in t-test (P < 0.01), while there was no difference between SMMC7721-mRz and SMMC7721-pEGFP-C1 (P > 0.05). With the advance of cell division, telomerase activities of the cells treated by SMMC7721-Rz and SMMC7721-mRz decreased gradually, and the percentage of apoptosis of the cells transfected with Rz and mRz increased gradually. The apoptosis percentage of 7PDS SMMC7721-Rz was 29.86%, while those of SMMC7721-mRz and SMMC7721-pEGFP-C1 were 9.87% and 3.36%, respectively.

CONCLUSIONThe apoptosis level of SMMC7721 induced by hTERT ribozyme increases as cells divide, and this ribozyme maybe a potential approach for liver cancer gene therapy.

Apoptosis ; physiology ; Cell Line, Tumor ; DNA-Binding Proteins ; genetics ; pharmacology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Mutation ; RNA, Catalytic ; genetics ; pharmacology ; Telomerase ; genetics ; pharmacology ; Transfection

This study was purposed to detect the expressions of CD271, CD133 and CD34, and to analyze the correlation of CD271 with CD133 and CD133 with CD34 expressions. The human bone marrow cells (BMCs) and mononucleated cells (MNCs) were detected by flow cytometry with CD45-PerCP, CD271-FITC, CD133-PE and CD34-FITC labelling according to different combinations of design, cells were located and selected repeatedly by FSC, SSC and CD45 after acquirement, then the expressions of CD271, CD133 and CD34 were detected by flow cytometry. The results showed that the expressions of CD271, CD133 and CD34 in BMCs were 0.16%, 0.20% and 0.43% respectively, while their expressions were 0.49%, 0.47% and 1.07% respectively after isolation of MNCs. The co-expressions of CD271(+)CD133(+) before and after isolation of MNCs were (0.02 +/- 0.01)% and (0.03 +/- 0.02)% respectively. The co-expression of CD133(+) and CD34(+) before and after isolation of MNCs were (0.18 +/- 0.11)% and (0.42 +/- 0.23)% respectively (p < 0.01); meanwhile about 90% of cells with CD133(+) expressed CD34 and 40% of cells with CD34(+) expressed CD133. It is concluded that the established method of detection using flow cytometry with three color fluorescence labelling can be used to detect expression of CD271, CD133 and CD34 in BMCs. The cells with CD271 are different from cells with CD133 and CD34, which suggests that the CD271 may be of important role in evaluating and guiding the clinical application of BM MSCs.
AC133 Antigen ; Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Bone Marrow Cells ; cytology ; metabolism ; Cell Line ; Flow Cytometry ; Glycoproteins ; metabolism ; Humans ; Nerve Tissue Proteins ; metabolism ; Peptides ; metabolism ; Receptors, Nerve Growth Factor ; metabolism

n increasing trend.

OBJECTIVETo immunopurify human endometrial endothelial cells (HEEC) from fresh surgical specimens of endometrial cancers and normal endometrial tissues, and investigate their biological characteristics.

METHODSEndothelial cells of endometrial cancers and normal endometrial tissues were isolated using anti-CD31 conjugated magnetic microbeads. The isolated endothelial cells were cultured in vitro and their origins were identified. Their angiogenic characteristics were observed by MTT, wound healing, Transwell cell invasion and tube formation assays.

RESULTSFlow cytometry revealed that the immunopurification technique yielded endothelial cell purity of > 95% in all samples. All purified HEEC were characterized as endothelial cells on the basis of expression of the classical endothelial markers vWF and CD31 as shown by immunofluorescence examination. Although the tumor-associated HEEC didn't show more rapid proliferation than normal HEEC, they exhibited enhanced migration ability (P = 0.006), potent invasiveness (P = 0.033), and elevated tube formation in vitro (P = 0.029).

CONCLUSIONSHuman endometrial endothelial cells can be efficiently isolated from endometrial cancer and normal endometrial tissues by immunomagnetic methods. Tumor-associated HEEC exhibit enhanced migratory ability, potent invasiveness, and elevated tube formation in vitro.

Adult ; Aged ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Endometrial Neoplasms ; metabolism ; pathology ; Endometrium ; cytology ; metabolism ; pathology ; Endothelial Cells ; metabolism ; pathology ; Female ; Humans ; Middle Aged ; Neoplasm Invasiveness ; Neovascularization, Pathologic ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; von Willebrand Factor ; metabolism

OBJECTIVETo investigate the clinical and pathological characteristics, treatment and prognosis of peripheral primitive neuroectodermal tumor (PNET) of the urinary tract and reproductive system.

METHODSThe clinical data and pathological characteristics of a PNET patient was analyzed and relevant literature reviewed.

RESULTSThe diagnosis was established by pathological and immunohistochemical method. The patient underwent radical surgery, followed by chemotherapy.

CONCLUSIONPathology and immunohistochemistry help the diagnosis of PNET. For the treatment of the tumors in the early stage, surgery is the best choice, and for that in the late stage, it can be followed by chemotherapy. The PNET of the penis is a rare disease and evidence still lacks for the evaluation of its prognosis.

12E7 Antigen ; Aged ; Antigens, CD ; analysis ; Cell Adhesion Molecules ; analysis ; Combined Modality Therapy ; Humans ; Immunohistochemistry ; Male ; Neuroectodermal Tumors, Primitive ; diagnosis ; metabolism ; therapy ; Penile Neoplasms ; diagnosis ; metabolism ; therapy ; Phosphopyruvate Hydratase ; analysis ; Prognosis