Child, Preschool ; Cystathionine beta-Synthase ; genetics ; Folic Acid ; blood ; metabolism ; Genetic Predisposition to Disease ; Heart Defects, Congenital ; enzymology ; genetics ; Humans ; Infant ; Infant, Newborn ; Methylenetetrahydrofolate Dehydrogenase (NADP) ; genetics ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Methyltransferases ; genetics ; Minor Histocompatibility Antigens ; Polymorphism, Genetic ; Risk Factors

Objective To establish a method to isolate and cultivate rat bone marrow mesenchymal stem cells (BMSCs),and explore the feasibility of labeling in vitro,and tracing in vivo,BMSCs with chloromethyl-benzamidodialkylcarbocyanine (CM-Dil). Methods BMSCs were obtained and subsequently cultured with whole bone marrow cell culture system,and the third generation of BMSCs was harvested. The positive rates of CD34,CD44 and CD29 were detected by flow cytometry. CM-Dil was used to label BMSCs in vitro and the efficiency of labeling after 24 hours and at day 21 and 30 were examined under fluorescent microscope. Moreover,the growth curves were sketched to determine the negative effects of CM-Dil on the vitality and proliferation of BMSCs cultured in vitro. Rat model of focal cerebral ischemia was reproduced,CM-Dil labeled BMSCs were implanted into corpus striatum of rats' brain with computer-guided stereotaxis thereafter. Brain tissues were obtained to prepare frozen sections at 7th,14th and 21st day post-implantation,and the survival and distribution of labeled cells were observed with fluorescent microscopy. Results The third generation passage of cultured BMSCs grew orderly,with shape of desmocytes,and homogeneous in morphology. The positive rates of CD34,CD44 and CD29 expressions in BMSCs were 1.71%,80.32% and 84.89%,respectively. Red fluorescence was observed in CM-Dil labeled BMSCs in vivo 24 hours after labeling,with a positive rate of 100%. The fluorescence intensity of passage cultured BMSCs observed on day 21 was similar to that observed at 24 hours after labeling,but diminished on day 30. The growth,proliferation and morphology of BMSCs,were not influenced by CM-Dil labeling. On day 7,14 and 21 post-implantation,BMSCs decreased in quantity,appearing in oval or irregular shapes,and most of the cells were found around the needle tract,with a tendency of diffusion to peripheral area. Conclusion High purity of BMSCs may be obtained with whole bone marrow cell culture system from bone marrow of rats. CM-Dil labeling is easy to handle and effective,with no cytotoxicity to cells. The latest labeling period of CM-Dil is 21 days in vitro,therefore it seems to be an effective method for in vivo tracing of BMSCs in brain after implantation.

Adolescent ; Catheter Ablation ; methods ; Humans ; Male ; Pre-Excitation, Mahaim-Type ; Tachycardia, Atrioventricular Nodal Reentry ; surgery

Objective Study the effect of NMDA receptor antagonist GAPA on the [Ca 2+ ]i of the bilirubin precipitated brain tissue. Methods 10 ug/g of GAPA was administrated peritoneally to Gunn rat with bilirubin induced encephalopathy, brain tissue suspensions was prepared, Fura 2/AM was loaded. the neural cytosolic Ca 2+ was measured by flurescence imaging analysis. Results The concentration of neural cytosolic Ca 2+ in bilirubin precipitated brain tissue was significantly more than that in the control group; NMDA receptor antagonist GAPA could significantly decrease the cytosolic Ca 2+ overload. Conclusion Cytosolic Ca 2+ overload was found in the bilirubin precipitated brain tissue. NMDA receptor antagonist GAPA could prevent the cytosolic Ca 2+ overload in bilirubin induced brain tissue.

Objective:To investigate if a plant-derived polysaccharide sulfate(M33A) binding to HIV1 gp120 may induce the exposure of neutralization epitopes of gp120,and if M33A-bound HIV-1ⅢB antigens may be used as a AIDS vaccine for inducing neutralizing antibodies against HIV-1.Methods:Whole-inactivated M33A-bound HIV-1ⅢB antigens were prepared and used to immunize mice after mixing with FCA or FIA.The titers of anti-HIV-1 IgG antibodies in immunized mouse plasma were detected by ELISA,and the HIV-1 neutralization by those plasma was detected by the improved microtiter neutralization assay.Results:M33A-bound HIV-1 antigens induced higher titers of anti-HIV-1 IgG antibodies(group C:1.5?10~6;group D:1.5?10~6) than HIV-1 antigens alone(4.9?10~5),and female mice produced 3 times higher titers of anti-HIV-1 IgG antibodies than male mice after immunized with various HIV-1 antigens.All three immunization schemes did not induce the production of anti-HIV-1 neutralizing antibodies.Conclusion:M33A binding does not induce gp120 to expose neutralization epitopes.However,M33A may improve the level of mice immune responses to HIV-1 antigens,suggesting M33A may enhance immune response to HIV-1 antigens.

Objective To assess the safety of emergent invasive treatment in severe postpartum hemorrhage. Methods Invasive treatment was performed in 18 patients with severe postpartum hemorrhage. The operation time, therapeutic effect, collateral circulation of uterus feeding arteries, pathological changes in uterine muscle after embolization and radiation dose received by the ovary during operation were investigated for safety assessment. Results (1) All eighteen patients who had been failed to conservative treatment were successfully cured with invasive treatment. The duration of hemostasis was 3 to 10 minutes, mean time (6?4) minutes. The operation time was 30 to 50 minutes, mean time (39?5) minutes. (2) Although both sides of internal iliac arteries or uterine arteries were embolized, a small amount of blood supply for uterus remains from ovary arteries, from branches external iliac arteries and from other arteries. (3) Uterine biopsies were performed in 3 patients 5 to 10 days after the invasive treatments for pathological examinations. Pathological findings showed that the necroses of uterine muscles were disconnected and the whole areas were blow 1/4. (4) The mean radiation dose received by the ovary in 5 cases was (17?7) cGy, which was within the safe limit. Conclusions The use of the emergency invasive treatment is safe in severe postpartum hemorrhage.

Objecttve To determine whether non-adherent bone marrow-derived mesenchymal stem cells (NA-BM-MSC) can differentiate into neuron-like cells in vitro and in vivo,especially in ischemic brain of mice. Methods NA-BM-MSC of mouse were indueed with conditioned medium containing epidermal growth factor and basic-fibroblast growth factor and demonstrated by immunocytochemistry for the expression of neuron specific nuclear protein and neurofilament-200. NA-BM-MSC from β-galactosidase transgenic mice constitutively expressing β-galactosidase were transplanted into the mice model of middle cerebral artery occlusion.LacZ staining and immunobistochemistry staining were performed to detect the survival,distribution and the differentiation of the donor cells in the ischemic brain. Results NA-BM-MSC can be induced into neuron specific nuclear protein and neurofilament-200 positive cells in vitro.LacZ staining showed that NA-BM-MSC can survive in ischemic brain and express β-galactosidase.Also,numbers of the neuron specific nuclear protein positive cells in β-galactosidase-positive cells were detected in the ischemic brain with double immunohistochemistry staining. Conclusions NA-BM-BMC can be induced to differentiate into neuron-like cells in vitro,and also can differentiate into neuron-like cells in ischemic brain tissue of the mice.

BACKGROUND: Triiodothyronine (T3) is an important regulation factor at the critical period of brain development. It maybe control the successive differentiation during the development of central nervous system (CNS).OBJECTIVE: To monitor the differentiation of neural stem cells (NSCs) induced by T3 and the thyroid hormone receptor (TR) mRNA expression changes.DESIGN: Open experiment.SETTING: Department of Pathology, Tianjin Medical College of Chinese People's Armed Police Force; Institute of Endocrinology of Tianjin Medical University.MATERIALS: This study was carried out in the Tianjin Medical University between January 2003 and March 2005.Ten-to-twelve-week-old aborted fetuses were obtained from the General Hospital of Tianjin Medical University with the approval of the local ethical committee. Informed consents were obtained from the mothers and their relatives.METHODS: ①Under the aseptic condition, the bilateral cortex of human fetal brain was removed and dissociated by brief mechanical trituration in D-Hanks. Then, 20 μg/L bFGF and 30 nmol/L T3 were used to induce the proliferation of NSCs and inoculated to poly-L-lysine-coated 24-well plate and 25 mL culture flask for routine culture at 1 ×109 L-1. The culture medium was DMEM/F12 serum-free complemented with N2. Half of the culture medium was changed every 48 hours.Seven days later, bFGF was discarded, only T3 was used for induction and differentiation. ② At 1, 2 and 3 weeks of culture, cells were collected, and RT-PCR was semiquantitatively used to detect TR mRNA expression changes at different stages of differentiation of NSCs. Isoforms were identified by immuocytochemistry.MAIN OUTCOME MEASURES: ①Cellular morphology observation and isoforms identification before and after differentiation of NSCs induced by T3. ② TR mRNA expression changes during the differentiation of NSCs.RESULTS: ①The hNSCs were round and had a smooth surface and gradually gathered to neurospheres. The proliferative hNSCs were nestin-positive and incorporated BrdU. When NSCs were induced by T3 for one week, most of the cells took on monopole or double poles, and had long and thin processes. The differentiated cells were neurofilament protein (NFP)-positive, galactocerebroside (GC)-positive or glial fibrillary acidic protein (GFAP)-positive. When NSCs were induced by T3 for three weeks, most of the cells were big, with unclear cell membrane, round nucleus, many thick processes which had many branches. The spider-like cells were scattered, and 80% of the cells were myelin basic protein-positive. ② TRα1 mRNA expression level was the highest before inducing NSCs. With the induction of T3, the expression level was decreased gradually, and was the lowest at 2 weeks, and then was rebounded gradually, but the final level was still lower than that of NSC (F =32.49, P =0.008). The tendency of TRα2 mRNA expression alteration was identical with that of TRα1 mRNA. TRβ1 mRNA expression level was the lowest in NSC, was increased gradually with the induction of T3 and attained the highest level at 2 weeks of induction of T3. Furthermore, the expression level of TRβ1 mRNA was also higher than that of TRα1 at the same time (t =15.64,P =0.001), and it reached the lowest level at 3 weeks of the induction. TRα3 expression level was firstly decreased after the differentiation induced by T3, and was close to the expression level of NSC at 2 weeks of induction (F =51.94, P =0.378), then was decreased to lower lever.CONCLUSION: T3 can induce NSC to differentiate into neurons, oligodendrocyte and astrocytes. TR mRNAs are expressed in different time intervals during the differentiation of NSCs.

Humans ; Hypertrophy ; diagnostic imaging ; Infant, Newborn ; Liver ; diagnostic imaging ; Male ; Tomography, X-Ray Computed

Animals ; Brain ; metabolism ; Codonopsis ; Excitatory Amino Acids ; metabolism ; Female ; Hypoxia ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Selenium ; pharmacology