Objective To study the influence of low-iodine diet on the expression of homeobox gene Nkx-6.1 and Nkx-6.2 in rat cerebrum tissue, and to explore the possible molecular mechanism of cerebrum development retardation caused by low-iodine. Methods Twenty female Wistar rats were randomly equally divided into two groups: low-iodine group and control group, both fed with low-iodine diet as low as 13.66 μg/kg determinated by spectrophotometry in Tianjin Institute of Endocrinology and the former with deionized water, the later 200 μg/L potassium iodate. Thyroid hormone level was detected using chemiluminescence immunoassay 3 months later and they were mated with male rats normally fed. Rats of 16-day pregnancy, new-born and 20th days old were detected the content of Nkx-6.1 and Nkx-6.2 mRNA in the cerebrum tissue by real-time fluorescence quantitative PCR 0.61), (3.28±0.80)pmol/L] were lower than the control group[(1.04±0.06), (39.42±14.68)nmol/L, (4.83±0.33), day pregnancy, new-born and 20th days old of control group was (1.90±0.23)×10-3,(1.86±0.40)×10-4, (1.11± 0.27)×10-4(F=827.58, P<0.01), Nkx-6.1 mRNA expression level gradually decreased along with aging(all P<0.05). The intra-group difference was significant (F=297.25, P<0.01) and the Nkxr.1 mRNA expression level during 16 days of pregnancy was the highest(P<0.01). It was higher in the control group than in the low-iodine group during 16 days of pregnancy (t=10.14, P<0.01) as well as in the low-iodine group than in the in 16-day pregnancy, new-born and 20th days old of control group was respectively(1.03±0.19)×10-2, (1.33± 0.10)×10-3, (8.79±0,87)×10-3, and that of low-iodine group was (0.31±0.03)×10-2, (1.53±0.13)×10-3, (7.51±0.86)×10-2. The intra-group difference was significant(F=1293.02,1065.83, all P<0.01). Nkx-6.2 expression level during 20th days old was the highest(P<0.01) and that of newborn was the lowest(P<0.01). The Nkx6.2 mRNA expression level in control group were higher than the low-iodine group during 16-day pregnancy and 20th days old(t=14.35, 4.05, all P<0.01). It was higher in the low-iodine group than in the control group during newboru(t=4.78, P<0.01). Conclusions The difference in the expression of Nkx-6.1 and Nkx-62 is highly related to the brain development retardation caused by low-iodine.

Objective To study the influence of low-iodine diet on the expression of homeobox gene nkx2.1 in rat cerebral tissue. Methods Forty female Wistar rats were randomly divided into two groups according to body. quality: low-iodine group and control group,both fed with low-iodine feed at an iodine content of 13.66 μg/kg,respectively given the deionized water and 200 μg/L KIO3 solution. The hormone levels of two group rats were determined with chemiluminescence immunoassay after three months, and then mated with healthy male rats. Cerebral tissues were taken from the fetus of 16-day pregnancy,newborn and 20 days old offspring in low-iodine and control group to detect the content of nkx2.1 mRNA using real-time fluorescence quantitative PCR (FQ-PCR) techniques. Results Serum TT3, TT4, FT3, FT4 level of rats in low-iodine group(0.89±0.20, 0.32±0.16, 3.33± 0.61, 3.28±0.80) was respectively lower than that in the control group(1.04±0.06, 39.42±14.68,4.83±0.33, 26.99±4.48;t = 2.71,6.52,5.70, 12.89, P < 0.05 or < 0.01). The relative nkx2.1 mRNA expression was(5.60± 0.30)×10-3, (1.20 ± 0.29)×10-3, (0.18± 0.06)×10-3 respectively in the fetus of 16-day pregnancy, newborn and 20 days old offspring of control group, while it was (3.00 ± 0.55)×10-3, (1.90 ± 0.21)×10-3,(0.69 ± 0.15)×10-3 in the low-iodine group. The difference of nkx2.1 mRNA expression was significant among fetal and neonatal rats in the control group and low-iodine group(F = 210.07,162.40, both P < 0.01). The nkx2.1 mRNA expression of newborn rats was lower than that of 16-day pregnancy in both groups(P < 0.01), and that of 20 days old rats was lower than that of 16-day pregnant and neonatal rats(P < 0.01). The 16-day pregnant rats of control group had obviously higher level of nkx2.1 expression than those in the low-iodine group(t = 16.073, P< 0.01), while the nkx2.1 of newborn and 20 days old low-iodine rats expressed much higher than healthy rats(t = 7.573,12.221, P < 0.01). Conclusions Brain development retardation caused by low-iodine is closely related to nkx2.1 differential expression in the brain tissue.

Objective:To explore the influence of low-iodine diet on the expression of homeobox gene NKX-2.2 in rat cerebrum tissue,and to explain the possible molecular mechanism of cerebrum development retardation caused by low-iodine.Methods:Female Wistar rats were randomly divided into two groups:Low-iodine group and control group,both fed with low-iodine feed,given the deionized water and KIO3 solution respectively,they were drawn from the 16-day pregnancy,new-born and 20th days old low-iodine and normal age offspring after three months,and detect the content of NKX-2.2 mRNA in the cerebrum tissue by Real-time fluorescence quantitative PCR techniques.Results:The thyroid hormone levels of low-iodine group in serum were significantly lower than the control group(P

OBJECTIVE: To observe the inductive efficiency of deriving hematopoietic cells from human embryonic stem (hES) cells co-cultured with human yolk sac stromal cells, fetal liver stromal cells or fetal bone marrow stromal cells,in order to discuss the effect of the different hemopoietic microenvironment on hemopoietic cytogenesis. METHODS: We used two-step method to induce the hES cells into the hematopoietic cells. In the first step the hES cells were co-cultured with cytokines by formation of the day 5 embryoid bodies (5d EBs). In the second step the 5d EB cells were induced into the hematopoietic cells by co-culturing with human yolk sac stromal cells, fetal liver stromal cells or fetal bone marrow stromal cells for 10 days. The inductive efficiencies of deriving hematopoietic cells from hES cells co-cultured with the different hemopoietic microenvironment were reflected by the expression levels of flk, CD34 and CD45 antigen. RESULTS: Flow cytometry analysis demonstrated that the population of the cells co-cultured with human yolk sac stromal cells contained flk (1.80%+/-0.56%), CD34 (1.30%+/-0.14%) or CD45 (1.05%+/-0.63%) positive cells; the population of the cells co-cultured with human fetal liver stromal cells contained flk (34.00%+/-25.45%), CD34 (38.40%+/-24.80%) or CD45 (72.60%+/-25.70%) positive cells; the population of the cells co-cultured with human fetal bone marrow stromal cells contained flk (2.50%+/-1.48%), CD34 (3.20%+/-0.56%) or CD45 (1.65%+/-0.21%) positive cells. Compared with spontaneous differentiation of EBs, all of the three stromal cells could induce EBs into the hematopoietic cells (P<0.05). CONCLUSION The inductive efficiency of deriving hematopoietic cells from EBs co-cultured with human fetal liver stromal cells was higher than EBs co-cultured with human yolk sac stromal cells and fetal bone marrow stromal cells.
Antigens, CD34 ; Cell Differentiation ; Cells, Cultured ; Cellular Microenvironment ; Coculture Techniques ; Embryonic Stem Cells ; cytology ; Fetus ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Leukocyte Common Antigens ; Mesenchymal Stem Cells ; cytology ; Stromal Cells ; cytology ; Yolk Sac ; cytology

OBJECTIVETo characterize the time course of spontaneous differentiation of in vitro cultured human embryonic stem cells (hESCs) into hematopoietic cells to provide experimental evidence for induction of hematopoietic commitment of hESCs.

METHODSIn human embryoid bodies (hEBs) derived from spontaneous differentiation of chESC3, a hESC cell line we established previously, the expressions of such genes as KDR, Bmi1, Scl and gata2 were detected by RT-PCR every other day during the 12-day differentiation to monitor the process of the hematopoiesis. The hematopoietic stem cell marker CD34 was examined using flow cytometry to evaluate the efficiency of hematopoietic differentiation of the cells on days 6, 8, 10 and 12. The spontaneously differentiated hESCs were seeded in the hematopoietic colony culture system to study the hematopoietic colony forming ability. Immunocytochemical staining for CD45 was performed on the hEBs to examine the emergence of mature hematopoietic cells.

RESULTSThe expressions of the hematopoietic stem cell-related genes KDR and Bmi-1 were detected in the hESCs, and on days 4 to 6, the two genes were upregulated with prolonged cuture of the hEBs. Scl and gata2 gene expressions were detected since 6-8 days of culture and maintained high expressions till day 12. Flow cytometry revealed a gradual increase in CD34-positive cells in the culture, with positivity rates on days 6, 8, 10, and 12 of (1.4-/+0.4)%, (3.4-/+1.3)%, (5.5-/+2.2)%, and (5.1-/+1.7)%, respectively. The numbers of CD43-positive cell colonies on days 6, 8, 10, and 12 were 0, 7-/+2, 37-/+11, and 89-/+29 in each 10(5) cells, respectively. Immunocytochemical staining identified CD45-positive cells on days 10, 12, 15, and 18 in the cell colonies, with the positive cell numbers of 0, 40.5-/+15.09, 178.6-/+55.89, and 253.0-/+52.04, respectively.

CONCLUSIONThe hESCs undergo spontaneous hematopoietic differentiation in 3 stages, including the differentiation into germ layer-specific cells (days 6-8), expansion period of the hematopoitic progenitors (days 8-12), and maturation of the hematopoietic cells (after day 15).

Animals ; Antigens, CD34 ; metabolism ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Cell Culture Techniques ; Cell Differentiation ; Embryonic Stem Cells ; cytology ; metabolism ; GATA2 Transcription Factor ; genetics ; Gene Expression Regulation ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; Mice ; Nuclear Proteins ; genetics ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; genetics ; Repressor Proteins ; genetics ; T-Cell Acute Lymphocytic Leukemia Protein 1 ; Time Factors

OBJECTIVETo investigate the effect of vascular bundle implantation in autogenous bone graft on angiogenesis.

METHODSThirty-six New Zealand white rabbits were evaluated in this study. A portion of bilateral radial bones of a rabbit were removed as free bone grafts, whose periostea were peeled off. In test group, the external maxillary artery bundle was passed through the marrow cavity of the bone. In control group, there was no vascular bundle implantation. Each bone was placed in masseter muscle separately. The rabbits were sacrificed and the specimens were procured at 3 days, 1, 2, 3, 4 and 6 weeks after surgery for histological observation, Chinese ink perfusion and CD34 immunohistochemistry. Microvessel density (MVD) was assessed in order to evaluate angiogenesis of autogenous bone grafts.

RESULTSThe bone grafts were found revascularization in 3 days after surgery in the test group, whereas at 2 weeks in the control group. In 3 days, 1 week, 2 weeks, 3 weeks and 4 weeks after surgery, the MVD of test group was significantly higher than that of control group. In 4 weeks after surgery, angiogenesis of test group reached to peak.

CONCLUSIONVascular bundle implantation improved angiogenesis in non-vascularized autogenous bone graft in this study.

Animals ; Bone Transplantation ; Bone and Bones ; Prostheses and Implants ; Rabbits

OBJECTIVETo study the magnetic resonance imaging (MRI) in the wrist joint of coal miners who work in excavation and vibration department.

METHODSForty-three coal miners with the hand-arm vibration disease served as the observation group while 20 workers who were not working in the vibration department acted as the control group. The patients in the observation group were divided into five subgroups according to the time when they received vibration. The regularity of the development of signs and symptoms of MRI was observed and analyzed.

RESULTSThe hydroarthrosis was most found in MRI. There were significant difference in hydroarthrosis (chi(2) = 8.80, P < 0.01), osteoporosis and osteomyelitis (chi(2) = 3.91, chi(2) = 5.01, P < 0.05 respectively) between the observation group and the control group. The edema of bone marrow and the avascular necrosis of ossa carpi were found only in the observation group and not found in the control group. The hydroarthrosis and the edema of bone marrow occurred most in the early stage of vibration. The signal in the edema of the bone marrow of the distal end of the radius was decreased in the GE sequence T(2)WI with the specificity.

CONCLUSION(1) Changes in the wrist joint occur in the early stage of the vibration work, and can be found in the MRI. (2) The edema of the bone marrow of the distal end of the radius is of great value in the diagnosis of the hand-arm vibration disease.

Adult ; Case-Control Studies ; Coal Mining ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Occupational Diseases ; diagnosis ; etiology ; Sensitivity and Specificity ; Vibration ; adverse effects ; Wrist Joint ; pathology

OBJECTIVETo clone a novel gene expressed specifically in human embryonic stem cell and to analyze its characteristics.

METHODSBased on an expression sequence tags(EST) CF948547 which expressed specifically in human embryonic stem cell, the full-length cDNA sequence of a novel gene was cloned by using bioinformatic and molecular biological technique. Its expression profile was analyzed by reverse transcription-polymerase chain reaction(RT-PCR), and subcellular location was determined by enhanced green fluorescent protein (EGFP) eukaryotic expression system.

RESULTSA novel gene HPESCRG1(homo sapiens pluripotent embryonic stem cell-related gene) was cloned successfully. Its GenBank accession number was AY283672. Its cDNA length was 1395 bp. It comprised 9 exons and 8 introns, and its opening reading frame was 250-1146 bp. Its chromosomal mapping was located in 3q13.13, and the putative protein contained 297 amino acids. The theoretical molecular weight of the putative protein was 33 784 and the isoelectric point was 9.35. The protein primary structure of this gene contained a SAP motif and it was subcellularly located in nuclei. Expression analysis showed that this gene was expressed specifically in human ES cells, but not expressed in the adult human tissues, the multiple tissues of embryo aborted in over 5 months' pregnancy, the differentiated cells of HESC-1, and the human mesenchymal stem cells (hMSCs) and human embryonic fibrocytes (hEFCs).

CONCLUSIONHPESCRG1 was found to be a novel gene expressed specifically in human ES cell, which might be related to self-renewal of human ES cell and maintaining its undifferentiated state.

Amino Acid Sequence ; Base Sequence ; Cell Line ; Cloning, Molecular ; DNA, Complementary ; analysis ; Embryo, Mammalian ; Exons ; Expressed Sequence Tags ; Gene Expression ; Humans ; Introns ; Molecular Sequence Data ; Molecular Weight ; Nuclear Proteins ; Open Reading Frames ; genetics ; Proteins ; genetics ; metabolism ; Stem Cells ; cytology ; metabolism

Objective To explore the correlation between induced abortion and reproductive tract infections (RTIs). Methods On the basis of keeping the representation of cities under study,53 652 fertile women aged 15-49 were surveyed by using a stratified-cluster-random sampling.Investigation and gynecological examination were conducted by two steps - firstly converging at the clinics, and then visiting those households for someone who did not show up at the clinics. Results Among all the 32.0% (n=16 800) women ever having experienced the history of induced abortion,21.1%(n= 11 090) of them had one, 7.6%(n=3976) women had two, and 4.1%(n=1734) women had at least three events. 59.0%(n=30 959) women among our studied samples had ever had RTI,with 30.9% ( n = 16 215 ) of them had only one 20.0% (n = 10 494 ) women had two and 8.1% (n =4250) had three or more RTIs. Data from x2 text and ordinal regression analysis revealed that the rural married women who underwent more induced abortions were more likely to suffer from RTIs,especially cervical infection and PID. Conclusion Our study showed that the rates of induced abortion and reproductive tract infections among married women in Anhui province were both high.Women who underwent induced abortions had a higher prevalence rate of reproductive tract infections.

AIM:To explore the regulatory effect of chemokine CCL 3 on exosome secretion from human bone marrow mesenchymal stem cells(hBMSCs).METHODS: hBMSCs were stimulated with chemokine CCL 3 at different concentrations in vitro.The proliferation of hBMSCs was measured by CCK-8 assay and viable cell counting.Exosome se-cretion from hBMSCs was qualitatively analyzed by transmission electron microscope(TEM)and flow cytometry, and the quantitative analysis was carried out by flow cytometry and nanoparticle tracking analysis(NTA).RESULTS:Compared with control group,the viability of the hBMSCs detected by CCK-8 assay was increased when hBMSCs were treated with CCL3(P<0.05).The results of viable cell counting demonstrated that the number of hBMSCs was raised in CCL 3 group in a dose-dependent manner(P<0.05).The results of flow cytometry showed that hBMSCs expressed 3 CCL3-related spe-cific receptors,CCR1,CCR5 and CCR9.Compared with control group,the fluorescence intensity of CCR9 in CCL3 group was obviously enhanced.However,no significant difference of fluorescence intensity for CCR 5 and CCR1 was observed be-tween the 2 groups.The results of NTA demonstrated that the secretion capacity of CCL 3-induced hBMSCs was far less than that in control group(P<0.05).However, the microvesicles larger than 100 nm in CCL3 groups were increased(P<0.05).The above results indicated that the higher concentration of CCL 3 induced the lower secretion of exosomes.In addi-tion,the results of flow cytometry demonstrated that CCL 3-induced hBMSCs showed lower quantity of CD 9 +exosomes than those in control group(P<0.01).CONCLUSION:CCL3 promotes the proliferation of hBMSCs but depresses the secre-tion of exosomes in a dose-dependent manner.CCL3 affects the size distribution of exosomes and increases the number of nonfunctional microvesicles of larger than 100 nm in size.CCL3 induces the expression of CCR9 in hBMSCs.