Objective　To investigate the molecular regulation of G1 arrest of mouse thymocytes induced by ionizing radiation. Methods　Cell cycle was analyzed by flow cytometry (FCM) following staining of cells with proidium iodide. Fluorescent staining and flow cytometry analysis were employed for measurement of protein expression. Results　It was demonstrated that G1 phase of mouse thymocytes increased significantly at 12h after whole body irradiation (WBI) with the doses of 0.5, 1.0 and 2.0 Gy, and at 24h following 2.0Gy exposure, measured by FCM. In the time course experiment, it was found that G1 phase of thymocytes increased significantly at 4h, reached a peak level at 24h and came down toward 48h after WBI with 2.0Gy X-rays. The results also showed that after 2.0Gy exposure, the expression of proteins in mouse thymocytes increased significantlty from 1h to 8h for p53, for p21 from 4h to 48h, and for MDM2 at 4h and 8h, measured by FCM. But no change was found for GADD45 protein expression. Conclusion　These results suggest that G1 arrest could be induced by a single dose of 0.5 Gy, 1.0Gy or 2.0Gy, and its molecular control might be established through the p53-p21 pathway.

Aged ; Drug-Related Side Effects and Adverse Reactions ; blood ; therapy ; Emergency Treatment ; Female ; Hemoperfusion ; Humans ; Hypnotics and Sedatives ; poisoning ; Male ; Middle Aged ; Serum ; chemistry

OBJECTIVETo evaluate the growth-inhibiting and pro-apoptotic effect of kaempferol in human esophageal squamous carcinoma Eca-109 cells and explore the mechanism.

METHODSThe effect of kaempferol on Eca-109 cell proliferation in vitro was measured by MTT assay. TUNEL staining was used to detect the cell apoptosis following kaempferol treatment. The changes in Bax and Bcl-2 mRNA expressions in response to kaempferol treatment were determined by RT-PCR, and the caspase-3 and caspase-9 activities were evaluated using colorimetric assay.

RESULTSKaempferol significantly inhibited Eca-109 cell proliferation (P<0.05) in a concentration-dependent manner and induced obvious cell apoptosis. RT-PCR showed that after kaempferol treatment caused up-regulated Bax and down-regulated Bcl-2 mRNA expression. The colorimetric assay revealed significantly increased caspase-3 and caspase-9 activities in Eca-109 cells following kaempferol treatment (P<0.01).

CONCLUSIONKaempferol can induce apoptosis of Eca-109 cells via a mitochondria-dependent pathway.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Squamous Cell ; pathology ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Esophageal Neoplasms ; pathology ; Humans ; Kaempferols ; pharmacology ; Mitochondria ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

The clinical and hematologic features in 22 patients with metastatic carcinoma of bone marrow were observed and analyzed. Morphology of bone marrow cells, bone marrow biopsy and other accessory examinations were performed. The primary or cardinal symptoms of metastatic carcinoma of bone marrow included anemia (17 cases, 77.3%), ostealgia (10 cases, 45.5%), fever (8 cases, 36.4%), hemorrhage (4 cases, 18.2%) and complicated hemolytic anemia (4 cases, 18.2%). The primary carcinomas, diagnosed by pathologic and accessory examinations, include gastric carcinoma (6 cases, 27%), lung cancer (3 cases, 13.6%), ovarian cancer (2 cases, 9%), mammary cancer, prostatic carcinoma, osteocarcinoma and metastatic malignant melanoma (1 case, respectively), and unknown primary lesion (7 cases, 31.8%). The hematologic features were decrease of hemoglobin (17 cases, 77.3%) and blood plate count (16 cases, 72.7%), leukocytosis (11 cases, 50%), immature leukocytes (14 cases, 63.6%) and erythrocytes (9 cases, 40.9%) seen on the peripheral blood smear, and reticulocytosis (4 cases, 18.2%). Masses of metastatic carcinoma cells can be frequently seen at two sides and tail of bone marrow smear. Bone marrow biopsy of 8 cases demonstrated the infiltration of carcinoma cells with nest-like distribution in the bone marrow cavity. Examination of MRI in 6 case showed destruction of bone and corpus vertebra and abnormal signal focus. Bone marrow biopsy could contribute to improve the accuracy of diagnosis and determine the origin of primary carcinoma. MRI plays an important role in diagnosis of metastatic carcinoma in bone marrow.

OBJECTIVETo investigate the significance of Th17/Treg imbalance in the development and treatment of primary immune thrombocytopenia (ITP) in children.

METHODSThirty-two children diagnosed with ITP between May and August, 2015 and 22 healthy children were enrolled. Flow cytometry was used to determine the Th17/Treg ratio in peripheral blood of healthy children and children with ITP before and after treatment with immunoglobulin.

RESULTSCompared with the patients with ITP before treatment, the healthy children and the patients treated with immunoglobulin had a significantly lower percentage of Th17 cells in CD4+ T cells, a significantly lower Th17/Treg ratio, and a significantly higher percentage of Treg cells in CD4+ T cells in peripheral blood (P<0.05). In the 32 ITP children treated with immunoglobulin, 20 had complete response, 4 had response, and 8 had no response. The patients with complete response had a significantly lower percentage of Th17 cells in CD4+ T cells and a significantly lower Th17/Treg ratio in peripheral blood than the patients without response (P<0.05).

CONCLUSIONSThe Th17/Treg imbalance can be found in children with ITP. Immunoglobulin can improve the cellular immune function by regulation of the Th17/Treg ratio. The Th17/Treg ratio may serve as an indicator for assessing the therapeutic effects of ITP.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology

OBJECTIVETo investigate the effect of ionizing radiation on the expression of p16, CyclinD1, and CDK4 in mouse thymocytes and splenocytes.

METHODSFluorescent staining and flow cytometry analysis were employed for the measurement of protein expression.

RESULTSIn time course experiments, it was found that the expression of p16 protein was significantly increased at 8, 24, and 48 h for thymocytes (P < 0.05, P < 0.01, and P < 0.05, respectively) and at 24 h for splenocytes (P < 0.05) after whole body irradiation (WBI) with 2.0 Gy X-rays. However, the expression of CDK4 protein was significantly decreased from 8 h to 24 h for thymocytes (P < 0.05-P < 0.01) and from 8 h to 72 h for splenocytes (P < 0.05-P < 0.01). In dose effect experiments, it was found that the expression of p16 protein in thymocytes and splenocytes was significantly increased at 24 h after WBI with 1.0, 2.0, and 4.0 Gy (P < 0.05-P < 0.01), whereas the expression of CDK4 protein was significantly decreased with 2.0 Gy for thymocytes (P < 0.05) and 0.5-6.0 Gy for splenocytes (P < 0.05-P < 0.01). Results also showed that the expression of CyclinD1 protein decreased markedly in both thymocytes and splenocytes after exposure.

CONCLUSIONThe results indicate that the expression of p16 protein in thymocytes and splenocytes can be induced by ionizing radiation, and the p16-CyclinD1/CDK4 pathway may play an important role for G1 arrest of thymocytes induced by X-rays.

Animals ; Cyclin D1 ; biosynthesis ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; Cyclin-Dependent Kinases ; biosynthesis ; Dose-Response Relationship, Radiation ; Flow Cytometry ; Male ; Mice ; Mice, Inbred Strains ; Proto-Oncogene Proteins ; Radiation Dosage ; Spleen ; cytology ; metabolism ; radiation effects ; Thymus Gland ; cytology ; metabolism ; radiation effects ; X-Rays

OBJECTIVETo investigate the effect of small hairpin interfering RNA (shRNA) in suppressing cytochrome P450 3A4 (CYP3A4) gene expression in CHL-3A4 cells.

METHODSThree shRNA expression vectors targeting CYP3A4 gene (CYP3A4 I, C YP3A4 II, and CYP3A4 III, respectively) were designed, synthesized and transfected into CHL-3A4 cells via liposomes. The inhibitory effect of shRNA on CYP 3A4 gene expression was detected by Western blotting and RT-PCR, and the effect of shRNA transfection in suppressing cyclophosphamide-induced cytotoxicity was measured using MTT assay.

RESULTSThe vector carrying CYP3A4 III shRNA significantly reduced the expression of CYP3A4 gene at both the mRNA (75%) and protein levels (80%) in CHL3A4 cells. The cytotoxicity of cyclophosphamide was markedly inhibited by CYP3A4 III-mediated suppression of CYP3A4 gene expression by 75% in CHL-3A4 cells.

CONCLUSIONThe vector-mediated RNA interference can suppress CYP3A4 gene expression in CHL-3A4 cells, and RNA interference technique provides a new means for studying cytochrome P450 gene function in mammalian cells.

Animals ; Cells, Cultured ; Cricetinae ; Cytochrome P-450 CYP3A ; genetics ; Fibroblasts ; cytology ; metabolism ; Humans ; Lung ; cytology ; RNA Interference ; RNA, Small Interfering ; genetics

OBJECTIVETo observe the effect of ketoconazole on the activity of cytochrome P450 (CYP) 1A2 and 3A4 in hepatic microsomes of healthy adults.

METHODSHuman hepatic microsomes obtained from healthy adults were randomly divided into control group and ketoconazole-treatment groups at different concentrations. After 15 min of culture, the substrates (testosterone for CYP3A4 and phenacetin for CYP1A2) were added and incubated for another 20 min. The metabolites (6-testosterone and acetaminophen) were then measured with high-performance liquid chromatography (HPLC) to assess the activities of CYP3A4 and 1A2.

RESULTSSignificant difference was found between the groups in the quantity of 6-testosterone and the relative activity of CYP3A4 (P<0.05). The IC(50) of ketoconazole for CYP3A4 was 0. 16 mg/L. Both the quantity of 6-testosterone and the relative activity of CYP3A4 were reduced gradually with the increment of ketoconazole concentration. Significant differences were found between the ketoconazole groups and the control group in both the quantity of acetaminophen and the relative activity of CYP1A2 (P<0.05). Ketoconazole at low doses reduced CYP1A2 activity and but increased the activities at high doses (P<0.05).

CONCLUSIONIn the range of maximum clinical blood concentration, ketoconazole can inhibit the activity of CYP3A4, but not that of CYP1A2, in the hepatic microsomes in healthy adults.

Adult ; Antifungal Agents ; pharmacology ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 CYP3A ; metabolism ; Dose-Response Relationship, Drug ; Female ; Humans ; Ketoconazole ; pharmacology ; Male ; Microsomes, Liver ; drug effects ; enzymology

BACKGROUNDIn 2008, infants in some areas of China suffered from stones of the urinary system which were caused by melamine-contaminated milk formula. Most of the infants were asymptomatic, and a few suffered from acute renal failure induced by urinary obstruction by stones. This study aimed to assess the significance of blood purification therapy in treatment of infants with acute obstructive oligo-anuric renal failure. Corrective perception, timely diagnosis, and active treatment of this complex disease are critical factors that guarantee a quick recovery of renal function of infants and help them to prevent multiple organ system failure.

METHODSThirteen infants with acute renal failure induced by urinary multiple obstruction caused by melamine-containing stones who had been admitted to Beijing Children's Hospital Affiliated to Capital Medical University in 2008 were investigated for the epidemiological characteristics, image features and indications of dialysis. All these infants were treated with dialysis. The efficacy of dialysis was compared with that of two control groups treated with cystoscopic retrograde catheterization into the ureter and medical treatment for the recovery of renal function.

RESULTSThe 13 infants with life-threatening complications treated with dialysis showed a blood urea nitrogen (BUN) level of (30.9+/-7.9) mmol/L and a creatinine (Cr) level of (572+/-173) micromol/L. Of these infants, 8 were treated with peritoneal dialysis (PD), and 5 with hemodialysis (HD). Ten infants recovered to urinate 24-72 hours after dialysis and 3 infants with persistent ureteral obstruction were further treated with cystoscopic retrograde catheterization into the ureter for drainage, and urination resumed soon after the operation. The average time of PD and HD were (2.1+/-0.8) days and (1.2+/-0.4) days, respectively. The total average time of PD and HD dialysis was (1.77+/-0.83) days. The recovery time of renal function of infants after dialysis was (3.08+/-1.20) days, comparable to that of the two control groups treated with catheterization with a cystoscope or by medication. There was no significant difference in the recovery time of renal function among the three groups (P>0.05).

CONCLUSIONSMelamine-contaminated milk formula may cause urinary stones and obstructive acute renal failure in infants. Dialysis is suggested to treat life-threatening complications such as hyperkalemia, oliguria or anuria if surgical intervention fails. If possible, hemodialysis or peritoneal dialysis can be performed early. Blood purification is feasible to help the infants overcome the critical stage of acute renal failure. Surgical measures can be taken to remove the obstruction if necessary.

Acute Kidney Injury ; chemically induced ; surgery ; therapy ; Child, Preschool ; China ; Cystoscopy ; methods ; Female ; Humans ; Infant ; Kidney Function Tests ; Male ; Peritoneal Dialysis ; methods ; Renal Dialysis ; methods ; Treatment Outcome ; Triazines ; poisoning

OBJECTIVETo explore the expression of MICA/B in human esophageal cancer, and to analyze its correlation with clinicopathological features.

METHODSThe expression of MICA/B in 40 cases of esophagus carcinoma and corresponding normal esophageal mucosa tissues were examined by immunohistochemistry.

RESULTSThe positive rate of expression of MICA/B protein in the esophageal carcinoma was 75.0% (30/40), and that in the corresponding normal esophageal mucosa was 0 (0/40). Up-regulation of MICA/B expression was found in the esophageal carcinomas. The expression of MICA/B was related with histological grade of the esophageal carcinoma (P = 0.012).

CONCLUSIONMICA/B protein plays an important role in the esophageal carcinogenesis, and my become a useful molecular marker for the diagnosis of esophageal carcinoma.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; metabolism ; Esophageal Neoplasms ; diagnosis ; metabolism ; pathology ; Female ; Histocompatibility Antigens Class I ; metabolism ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Grading ; Up-Regulation