AIMTo establish model of differentiation of fetal liver stem cells induced by beta-ME + BHA into neural cells in vitro;

METHODSCD34+ cells from naturally aborted human fetal liver were isolated with MACS Kit, and cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS). After confluent more than 80%, the 5 passage cells were induced by 10(-3) mol/L beta-mercaptoethanol (beta-ME) and 2 x 10(-4) mol/L BHA for 24 hours, and washed with PBS, and then incubated in serum-free medium for 5 hours to 5 days. The characteristics of treated cells were assayed by immunocytochemistry staining analysis.

RESULTSCells treated by beta-ME+ BHA exhibited neuronal phenotype, and expressed neuronal specific proteins such as nestin, NeuN, TrnJ-1, and NF-M, which were not found in control cells. Statistic analysis showed that 81% cells were NeuN-positive, 75% cells TuJ-1-positive, 47% cells NF-M-positive, 90% cells NSE-positive.

CONCLUSIONbeta-ME + BHA can induce human fetal liver CD34+ cells to produce neuronal specific antigens and proteins in vitro and become neuronal cells. CD34+ cells from human fetal liver possess potentials of differentiation into neural cells.

Antigens, CD34 ; Butylated Hydroxyanisole ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Liver ; cytology ; embryology ; Mercaptoethanol ; pharmacology ; Neurons ; cytology

Actins ; metabolism ; Carcinoembryonic Antigen ; metabolism ; Desmin ; metabolism ; Diagnosis, Differential ; Epithelial Cells ; metabolism ; pathology ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Neoplasms, Complex and Mixed ; metabolism ; pathology ; surgery ; Neoplasms, Glandular and Epithelial ; metabolism ; pathology ; surgery ; Stromal Cells ; metabolism ; pathology ; Vimentin ; metabolism

To determine the role of allogeneil graft of mesenchymal stem cells in mammalian longevity, mesenchymal stem cells were isolated from BALB/c mouse uterine-incision delivery fetus by two successive density gradient centrifugations, and then were purified and amplified by adherent culture. Identified P1 mesenchymal stem cells were injected (i.v.) through vena caudalis into the 15-month-old female BALB/c mice three times. The mice were evaluated with ultrasoundcardiogram, autopsy, score of cardiac, skin, lung, kidney, colon histopathology and serum total superoxide dismutase activity, maleic dialdehyde content, glutathione peroxidase activity. The results showed that after transplantation, the long-term surviving stem cells were found to be located in many organ tissues with in situ Y chromosomal hybridization dyeing. Median life span was increased in these animals after transplantation. Skin, cardiac, lung, kidney and colon pathology development were delayed. The retrogradation of heart function was attenuated, the increase of heart mass index (the mass of heart/the mass of the body), and serum maleic dialdehyde content, the decrease of spleen mass index (the mass of spleen/the mass of the body), serum total superoxide dismutase activity and glutathione peroxidase activity were reduced three months after transplantation (all P<0.05). These results support the idea that longevity can be enhanced by transplantation of mesenchymal stem cells and reinforce the hypothesis of mesenchymal stem cell as antiager.
Aging ; physiology ; Animals ; Female ; Fetal Stem Cells ; transplantation ; Longevity ; physiology ; Mesenchymal Stem Cell Transplantation ; Mice ; Mice, Inbred BALB C ; Random Allocation

AIMTo study the mechanism of butylated hydroxyanisole-induced neural differentiation of fetal liver cells in vitro.

METHODS14.5-day-old mouse fetal liver-derived cells were cultured, and were induced by 200 micromol/L butylated hydroxyanisole (BHA) combined with PI3K inhibitor LY294002 (20 micromol/L), and then were incubated in serum-free medium. Expression of genes in treated or untreated cells were assayed by Western blotting or RT-PCR.

RESULTSThere was low level of neurofilament-L (NF-L) and brain factor-1 (BF-1) but no neurofilament-H (NF-H) and tyrosine hydroxylase (TH) in fetal liver cells. BHA promoted significantly expression of neuron-specific NF-L, NF-H, BF-1, and TH in fetal liver cells. NF-L mRNA increased 5.8 fold, NF-H mRNA 8.0 fold, BF-1 mRNA 2.68 fold, and TH mRNA 30 fold, respectively (all P < 0.01 vs untreated cells). NF-L protein increased 11.29 fold, NF-H 5.5 fold, BF-1 2.53 fold, TH 4.76 fold. Moreover, expression of these BHA-induced genes were inhibited by PI3K inhibitor LY294002.

CONCLUSIONBHA induced neural differentiation of fetal liver cells through PI3K.

Animals ; Butylated Hydroxyanisole ; pharmacology ; Cells, Cultured ; Embryo, Mammalian ; cytology ; Hepatocytes ; drug effects ; metabolism ; Mice ; Mice, Inbred C57BL ; Nerve Tissue Proteins ; biosynthesis ; genetics ; Phosphatidylinositol 3-Kinases ; metabolism

In order to determine the involvement of CALM-AF10 fusion transcripted in primary leukaemias with t(10;11) and its chemotherapy sensitivity in vitro, the AF10-CALM fusion transcripts were detected by reverse transcription-polymerase chain reaction (RT-PCR), and the chemotherapy sensitivity testing in vitro was undergone by MTT assay in five t(10;11) leukemia samples from patients with ALL, AML and lymphoblastic lymphoma. The results showed that five different-sized AF10-CALM product and four different-sized CALM-AF10 products were detected. The chemotherapy sensitivity of leukemic cells with t(10;11) in vitro to drugs is lower than that of leukemic cells without t(10;11). 3 out of 5 cases of t(10;11) leukemia were sensitive to chemotherapeutic drugs, while 31 out of 36 cases of leukemia without t(10;11) were sensitive at same condition. There were significant differences (P < 0.01), consistent with clinical features of patients. Apoptosis rate of leukemic cells with t(10;11) induced by chemotherapeutic drugs was lower than that of leukemic cells without t(10;11), (16.37 +/- 2.56)%, and (33.75 +/- 5.59)%, respectively (P < 0.01). It is concluded that the CALM-AF10 fusion transcripts are a common features and are involved in the pathogenesis of haematological malignancies with t(10;11), and are associated with a poor prognosis.
Antineoplastic Agents ; pharmacology ; Cell Survival ; drug effects ; Chromosomes, Human, Pair 10 ; genetics ; Chromosomes, Human, Pair 11 ; genetics ; Humans ; Leukemia ; genetics ; pathology ; Oncogene Proteins, Fusion ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transcriptional Activation ; drug effects ; Translocation, Genetic ; Tumor Cells, Cultured

OBJECTIVETo investigate the clinical features of thyroid disease occurring in response to antiviral therapy in patients with chronic hepatitis C (CHC).

METHODSEighty-two patients diagnosed with CHC were recruited for study from our hospital between 2009 and 2010. All patients were given a 48-week course of antiviral combination therapy with pegylated-interferon (Peg-IFN; 180 mug qw ih) and ribavirin (RBV; 15 mg/kg bw). Patient sera was collected prior to treatment (baseline), at treatment weeks 24 and 48, and post-treatment week 24, and used to detect changes in levels of thyroid function markers, thyroid-specific and other autoantibodies, complement factors, and immunoglobulins (Igs). Differential expression of biomarkers was assessed between patients who developed thyroid disorder and those who did not.

RESULTSAt treatment week 48, 13.4% (11/82) of cases developed hypothyroidism, 3.7% (3/82) developed hyperthyroidism, 20.7% (17/82) tested positive for thyroglobulin antibody, and 22.0% (18/82) tested positive for thyroid peroxidase antibody. The patients who did not develop thyroid disease had significantly higher post-treatment levels (vs. baseline) of IgG (14.84 +/- 2.61 vs. 12.95 +/- 3.32 g/L, F = 10.458, P = 0.002) and C4 (0.26 +/- 0.09 vs. 0.22 +/- 0.08 g/L, F = 6.835, P = 0.011) and significantly lower IgM (0.86 +/- 0.48 vs. 1.00 +/- 0.42 g/L, F = 9.106, P = 0.003). The patients who developed thyroid disease showed no significant differences in the baseline and post-treatment levels of IgG, C4, or IgM. When the two groups of patients who did or did not develop thyroid disease were compared, there was no difference in the amount of patients who achieved sustained virological response.

CONCLUSIONAntiviral-induced thyroid disease in patients with refractory hepatitis C manifests as clinically-detectable abnormalities in serum levels of thyroid autoantibody and markers of hypothyroidism. Levels of other autoantibodies and Igs do not correlate with the development of thyroid disease in these patients, and thyroid disease does not appear to affect the efficacy of Peg-IFN + RBV antiviral therapy.

Antiviral Agents ; therapeutic use ; Drug Therapy, Combination ; Hepatitis C, Chronic ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Polyethylene Glycols ; therapeutic use ; Ribavirin ; therapeutic use ; Thyroid Diseases ; chemically induced

OBJECTIVETo establish a stable rat experimental model of concealed penis for studying the effect of buried penis on the structure and function of the corpus cavernosum.

METHODSNinety male SD rats, aged 2 weeks, were randomly divided into 3 groups (A, B and C) of equal number. Groups A and B underwent surgery with intra-purse suture of the penile root and folding suture of the prepuce, respectively, to bury the penis, while Group C were included as sham operation controls.

RESULTSIn Group A, death resulted in 4 cases from acute post-operative urine retention, failure in burying the penis occurred in 5 cases because of soft tissue ulceration around the urethral orifice and in another 3 due to loose concealment. In Group B, 1 died from deep anesthesia and 2 from acute post-operative urine retention. With the penile development and erection, 7 in Group A and 10 in Group B protruded the penis in different stages. In Group C, 1 died from deep anesthesia. The operations succeeded in all the other rats in Groups A and B, with the success rates of 36.7% and 56.7%, respectively. And the concealment could be relieved any time during the experiment.

CONCLUSIONThe experimental rat model of concealed penis can be successfully established by both intra-purse suture of the penile root and folding suture of the prepuce, which is stable and similar to the natural course of this disorder in human.

Animals ; Disease Models, Animal ; Humans ; Male ; Penis ; abnormalities ; surgery ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Urogenital Abnormalities

OBJECTIVETo investigate the correlation between visceral adipose tissue-derived serine protease inhibitor (vaspin) concentration and insulin sensitivity in the visceral adipose tissue of young obese Sprague-Dawley (SD) rats.

METHODSTwenty-four SD rats which had been weaned 3 weeks before were randomly divided into two groups (n=12 each) to receive a high-fat and normal diet. The weight and abdominal circumference (AC) of each rat were measured, the fasting plasma glucose (FPG) and fasting insulin (FINS) in blood from the angular vein were measured after 12 hours of fasting and blood glucose (BG) and insulin (INS) levels in blood from the angular vein were measured at 60 and 120 minutes after intraperitoneal injection of 50% glucose (2 g/kg). The rats were sacrificed, and their liver and visceral adipose tissue were weighed. The vaspin concentration of the visceral adipose tissue in each rat was measured using ELISA. Correlation analysis was performed on the vaspin concentration and other indices.

RESULTSCompared with the normal diet group, the high-fat diet group showed significantly higher weight, AC, weight of visceral adipose tissue, FPG, FINS, 120 minute INS level, vaspin concentration, homeostasis model assessment for insulin resistance (HOMA-IR) and homeostasis model assessment of β cell function (HOMA-β) (P<0.05) Insulin sensitivity index (ISI) was significantly lower (P<0.01). Vaspin concentration was positively correlated with visceral adipose tissue and liver weight, AC, 120 minute INS level, FPG, FINS, HOMA-IR and HOMA-β (P<0.05), and negatively correlated with ISI (P<0.05).

CONCLUSIONSHigh expression of vaspin is associated with insulin resistance in young obese SD rats. Vaspin is presumably an adipocytokine that can increase insulin sensitivity, promote insulin secretion by islet β-cells and improve glucose tolerance, and it may be involved in insulin resistance and the disturbance of carbohydrate metabolism.

Animals ; Female ; Glucose Tolerance Test ; Insulin ; blood ; Insulin Resistance ; Intra-Abdominal Fat ; chemistry ; Male ; Obesity ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serpins ; analysis ; physiology

OBJECTIVETo investigate the effect of hTERT antisense oligodeoxynucleotide (ASODN) on the oncogenicity and the inductive apoptosis of HL-60 cells.

METHODSApoptosis of HL-60 cells was detected by flow cytometry (FCM) and agarose gel electrophoresis. Both treated and untreated HL-60 cells were collected and transplanted into 5 BALB/c nude mice respectively, the formation of transplanted neoplasm and its morphologic change were observed. After the transplanted neoplasms were uniform with the ameliorated method in another 10 BALB/c nude mice, they were divided into 2 groups and injected ASODN and PBS into the neoplasm respectively. Seven days later, the tumor were measured, its morphology were observed, and the apoptotic cells were detected with a TUNEL kit.

RESULTSAfter 72 h treatment there were DNA ladders and early apoptosis peak in hTERT ASODN treated HL-60 cells but was none in SODN treated and blank control cells. In tumor formation experiment, neoplasms were formed in ASODN treated group at 16-17 d and untreated group at 12-13 d. Neoplasm was formed in 2 of 5 ASODN treated mice and 4 of 5 untreated mice respectively. In untreated mice tumor tissues were rich in blood vasa and stromal tissue compared with that in ASODN treated mice. In tumor therapy experiment, before treatment, there was no difference in the average neoplasm physical volume between ASODN treated group [(100.9 +/- 24.6) mm3] and PBS treated group [(98.4 +/- 23.1) mm3] (P > 0.05). After treatment, the neoplasm volume in ASODN treated group [(422.7 +/- 326.4) mm3] was smaller than that in PBS treated group [(786.4 +/- 357.6) mm3] (P < 0.05). Histologically, there were many apoptosis cells in ASODN treated group, but was seldom seen in PBS treated group. The TUNEL positive cells in ASODN treated group were much more than that in PBS treated group (P < 0.05).

CONCLUSIONThe hTERT ASODN induces apoptosis of HL-60 cells in vitro, reduces the tumor formation in BALB/c nude mice and inhibits the growth of the transplanted neoplasm.

Animals ; Apoptosis ; drug effects ; HL-60 Cells ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Telomerase ; genetics ; Transfection ; Xenograft Model Antitumor Assays