HK-2 cells were cultured with various concentrations of glucose and insulin for 12,24,48,72 h.Transforming growth factor-?_1(TGF-?_1) protein in supematant was measured by ELISA,while TGF-?_1 mRNA expression was assessed by RT-PCR.Data showed that high concentration of glucose and insulin up-regulated the expression of TGF-?_1 in HK-2 cells through different pathways.

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.

Objective To investigate the gene expression profiles of peri-implantation endometrium of high responders during controlled ovarian hyperstimulation. Methods High responders with cancelled embryo-transfer during controlled ovarian hyperstimulation (high responder group, n=4) and healthy fertile volunteers (control group, n=3) were performed endometrial biopsies during peri-implantation. Histologic changes of endometrium were observed by HE staining, genes of differential expression were screened with microarrays Affymetrix U133A 2.0 and identified by Real-time PCR. The biological process analysis was performed by online biological information analysis tool PANTHER. Results The ehdometrium was in mid-secretory phase in control group, while development delay was found in some glandular organs in endometrium of high responder group. Three hundred and sixty-four genes of differential expression were screened, among which 233 were up-regulated genes and 131 were down-regulated genes. OPN, PLA2G2, DPPIV, IGFBP5 and SSAT were identified as endometrial function-related genes, whose Real-time PCR findings were positively correlated to gene signal values detected by microarray(r=0.44, P<0.01). PANTHER analysis indicated that genes of differential expression participated in the biological processes of cytokine signal transduction and immunological regulation. Conclusion Ovarian high response affects the gene expression profiles of peri-implantation endometrium, which may be one of the causes of sub-optimal endometrial receptivity.

This study examined associations between MTHFR C677T polymorphism and serum folate concentrations with the risk of esophageal precancerous lesions (EPL) and esophageal squamous cell carcinoma (ESCC). The highest quartile of serum folate concentration significantly decreased the risk of ESCC compared with the lowest quartile (OR=0.11; 95% Cl, 0.04-0.33; P<0.05). MTHFR 677 C>T polymorphism was associated with the risk of ESCC by using chi-square tests (P<0.05). For the CT genotype, the risk of ESCC significantly increased in study participants with low serm folate concentrations (≤26.92 μg/L) compared with participants with high serum folate concentrations (>26.92 μg/L) by using multinomial logistic regression models. The MTHFR genotype may further modify associations between serum folate concentrations and the risk of ESCC, but it was not significantly associated with the risk of EPL.
Carcinoma, Squamous Cell ; blood ; genetics ; Chi-Square Distribution ; Esophageal Neoplasms ; blood ; genetics ; Folic Acid ; blood ; Genetic Predisposition to Disease ; Humans ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Polymorphism, Genetic

OBJECTIVETo investigate the effect of X-rays on expression of caspase-3 and p53 protein in EL-4 cells and its implications in induction of apoptosis and polyploid cells.

METHODSMouse lymphoma cell line (EL-4 cells) was used. Fluorescent staining and flow cytometry analysis were employed for measurement of protein expression, apoptosis, cell cycle, and polyploid cells.

RESULTSThe expression of caspase-3 protein increased significantly at 8 h and 12 h, compared with that of sham-irradiated control (P<0.05, respectively) and the expression of p53 protein increased significantly at 2, 4, 8, 12, and 24 h, compared with that of sham-irradiated control (P<0.05-P<0.01) in EL-4 cells after 4.0 Gy X-irradiation. Apoptosis of EL-4 cells was increased significantly at 2, 4, 8, 12, 24, 48, and 72 h after 4.0 Gy exposure, compared with that of sham-irradiated control (P<0.05-P<0.001). G2 phase cells were increased significantly at 4, 8, 12, 24, 48, and 72 h (P<0.05-P<0.001). However, no marked change in the number of 8 C polyploid cells was found from 2 to 48 h after 4.0 Gy exposure.

CONCLUSIONThe expressions of caspase-3 and p53 protein in EL-4 cells are induced by X-rays, which might play an important role in the induction of apoptosis, and the molecular pathway for polyploid formation might be p53-independent.

Animals ; Caspase 3 ; metabolism ; radiation effects ; Caspases ; metabolism ; radiation effects ; Cell Line, Tumor ; Mice ; Tumor Suppressor Protein p53 ; metabolism ; radiation effects ; X-Rays

Apoptosis of abnormal oocytes is essential for defective oocyte elimination during prepubertal ovary development, and the ubiquitin system regulates the cell apoptosis via the degradation of specific proteins. Ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) is a component of the ubiquitin system, and the UCH-L1-dependent apoptosis is important for spermatogenesis. In the present study, the change in the number of follicles and the expression of UCH-L1 in oocytes were determined in prepubertal mouse ovaries by immunohistochemical techniques. A significant decrease in the follicular pool was found in prepubertal mouse ovaries during the period of day 21 to day 28 after birth, and accordingly, the UCH-L1 protein expression was increased, to some degree in association with Jun activation domain-binding protein 1 (Jab1) and cyclin-dependent kinase inhibitor p27(Kipl). The increased UCH-L1 protein, together with the corresponding changes of Jab1 was detected in morphologically abnormal oocytes of prepubertal ovaries. Through the immunofluorescent colocalization, UCH-L1 was shown concentrating in abnormal oocytes, and a parallel change in Jab1 was also seen. The affinity analysis confirmed the interaction between UCH-L1 and Jab1 in ovaries. These results suggest that UCH-L1 plays an important role, possibly in association with Jab1 and p27(Kipl), in selective elimination of abnormal oocytes during mouse prepubertal development.
Animals ; Apoptosis ; Female ; Mice ; Oocytes ; cytology ; Ovary ; enzymology ; Ubiquitin Thiolesterase ; metabolism

OBJECTIVETo study the combined toxic effects of fumonisin B(1) (FB(1)) and aflatoxin B(1) (AFB(1)) on Sprague-Dawley (SD) rats.

METHODAll 60 SD male rats were divided into five groups randomly according to the body weight (12 every group). They were given FB(1) (100 microg/kg bw), AFB(1) (100 microg/kg bw), FB(1) plus AFB(1) (100 microg/kg bw respectively), FB(1) plus AFB(1) (50 microg/kg bw respectively) and distilled water respectively by gavage. The experiment persisted 30 days to observe the changes of growth and development, the food used rate, the haematological indexes, the blood biochemical indexes and the viscera histopathology.

RESULTSAt the end of the experiment, the mean body weight increased in the FB(1) plus AFB(1) (100 microg/kg bw respectively) group was (164.9 +/- 19.8) g and the mean body weight increased in the control group was (203.7 +/- 17.1) g. And the food used rate in the FB(1) plus AFB(1) (100 microg/kg bw respectively) group was (25.3 +/- 1.6)% and the food used rate in the control group was (28.1 +/- 1.2)%. There were significant differences in the mean body weight increased and the food used rate between the FB(1) plus AFB(1) (100 microg/kg bw respectively) group and the control group (P < 0.05). While there were no significant differences of body weights and food used rates between controls and AFB(1), FB(1), and low dose AFB(1) + FB(1) groups (P > 0.05). The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutaminetransferase (gamma-GT) in serum of all of the treatment groups were increased, but the increasing extent was severe in the AFB(1) + FB(1) high dose group. At the same time the liver weight and kidney weight were decreased and the liver occurred with the remarkable histopathological lesions in the AFB(1) + FB(1) high dose group. The activity of superoxide dismutase (SOD) in serum was decreased and the level of malondialdehyde (MDA) in serum was elevated in treatment groups.

CONCLUSIONSThe combined toxic effects of AFB(1) and FB(1) existed in male SD rats. Our results provided the basic data for studying the combined effects on human exposed to these two mycotoxin at the same time.

Aflatoxin B1 ; toxicity ; Animals ; Fumonisins ; toxicity ; Male ; Mycotoxins ; toxicity ; Rats ; Rats, Sprague-Dawley ; Toxicity Tests

Nine compounds were isolated from an ethanol extract of the roots of K. roxburghii by using a combination of various chromatographic techniques including column chromatography over silica gel, MCI gel, Sephadex LH-20, and reversed-phase HPLC. On the basis of physical-chemical properties and spectroscopic data analysis, their structures were identified as munjistin (1), 1-methoxy-3,6-dihydroxy-2-hydroxymethyl-9,10-anthraquinone (2), 1,2,3-trihydroxy-9,10-anthraquinone (3), arjunolic acid (4), hyptatic acid-A (5), hyptatic acid-B (6), 2α,3β,24-trihydroxyurs-12-en-28-oic acid (7), 2α,3β,23-trihydroxyurs-12-en-28-oic acid (8), and daucosterol (9). Compounds 1-9 were obtained from this genus for the first time.
Anthraquinones ; chemistry ; isolation & purification ; Rubiaceae ; chemistry ; Triterpenes ; chemistry ; isolation & purification

OBJECTIVETo evaluate cord blood stem cell transplantation (CBT) in the treatment of X-linked agammaglobulinemia, and observe the courses of the hematopoietic and immune reconstitution.

METHODSA 14-year-old male patient with agammaglobulinemia received CBT from a 1/6 HLA-mismatched unrelated cord blood. The conditioning regimen was Bu/Cy/anti-CD3 antibody. CsA was given together with MMF and MTX for prophylaxis of GVHD. The patient received 0.42 x 10(8) nucleated cells/kg, containing 0.35 x 10(6) CD34(+) cells/kg.

RESULTSThe recipient showed hematopoietic reconstitution on day 30 post-transplantation when ANC was 0.5 x 10(9)/L and BPC 20 x 10(9)/L. Sex chromosome analysis showed engraftment (donor 46, XX/recipient 46, XY = 4:1) on day 45. The recipient's blood group changed from AB to O, IgG from 1.1 g/L to 3.5 g/L, sex chromosome from 46, XY to full 46, XX, and mature B cells in peripheral blood from 0 to 5% on day 100, indicating immune reconstitution. At the last follow-up of 360 days, the patient without acute or chronic GVHD showed normal hemogram and myelogram, IgG 13.5 g/L and 10% mature B cells in peripheral blood, indicating the hematopoiesis and immune persistent reconstitution. No acute or chronic GVHD was developed.

CONCLUSIONThis is the first case report of successful treatment of X-linked agammaglobulinemia by HLA-mismatched unrelated CBT.

Adolescent ; Agammaglobulinemia ; surgery ; Cord Blood Stem Cell Transplantation ; methods ; Graft vs Host Disease ; prevention & control ; HLA Antigens ; immunology ; Humans ; Male ; Transplantation Conditioning ; Treatment Outcome

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