Objective To study the effect and mechanism of indomethacin(IN) on protein maps of human colon cancer cell line (HCT116). Methods Proteome profiles of HCT116 in IN treated and control group were separated by immobilized pH gradient based two dimensional gel electrophoresis (2 DE). Differential expression of protein and proteome analysis were identified and analyzed by silver scan using Image Scanner and ImageMaster software. Peptide mass fingerprint (PMF) was based on matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) and database searching. Results Clear background, well resolution and reproducible 2 DE patterns of HCT116 cells were acquired in IN treated and control group. The protein spots and match rate in control group and IN treated group were 1299?55 (94.2%) and 1208?47 (91.0%) . There were totally 45 differential protein spots, of which the expression of protein in IN treated group was down regulated in 34 spots and up regulated in 9 spots, while 2 spots of protein was expressed only in control group. Twelve differential proteins were identified by PMF, and HCT116 cells apoptosis was through BfL 1 signaling pathway. Conclusion IN can induce HCT116 cells apoptosis though BfL 1 signaling pathway.

Objective To determine the value of MR in the diagnosis of the medial and lateral meniscal posterior root tears.Methods One hundred and twenty one patients underwent arthroscopy and MR of knee examinations were retrospectively reviewed,and all of them did not have the history of knee surgery before the MR examination.MR images were double-blinded,independently,retrospectively scored by radiology residency (doctor 1) and attending radiologist (doctor 2).The likelihood of the meniscus posterior root tear was scored on basis of arthroscopic findings constituted the gold standard.Sensitivity,specificity and accuracy of MR diagnosis of the lateral and medial meniscus posterior root tear by two physicians were calculated,and the consistency of diagnosis results by two physicians was evaluated using the Kappa statistics.Reasons leading to misdiagnosis or missed diagnosis were discussed.Results Thirty three of the 121 patients were diagnosed with meniscus posterior root tears by arthroscopy,including 14 cases of lateral tears and 19 cases of medial tears.The sensitivity,specificity and accuracy in the diagnosis of posterior lateral meniscus root tears (PLMRT) for doctor 1 were 71.4％ (10/14),82.2％ (88/107) and 81.0％ (98/121),respectively,and for doctor 2 were 71.4％ (10/14),86.0％ (92/107) and 84.3％ (102/121).The sensitivity,specificity and accuracy in the diagnosis of posterior medial meniscus root tears (PMMRT) for doctor 1 were 100.0％ (19/19),88.2％ (90/102) and 90.1％ (109/121),respectively,and for doctor 2 were 100.0％ (19/19),95.1％ (97/102) and 95.9％ (116/121),respectively.The Kappa statistics for PLMRT and PMMRT were 0.67 and 0.81,respectively,demonstrating a good interobserver agreement (P＜0.05).The misdiagnosed cases of the PMMRT were 12 for doctor 1 and 5 for doctor 2.There were 4 cases being commonly misdiagnosed by the two doctors,and there were no any missed cases in this study.For the PLMRT,the misdiagnosed cases were 19 for doctor 1 and 15 for doctor 2.Eleven cases were misdiagnosed and 4 were missed by the two physicians.Conclusion MR is a reliable diagnostic tool for detecting posterior root tears of the medial and lateral meniscus.

9α-hydroxy-4-androstene-3,17-dione (9-OH-AD) is an important intermediate in the steroidal drugs production. 3-ketosteroid-9α-hydroxylase (KSH), a two protein system of KshA and KshB, is a key-enzyme in the microbial steroid ring B-opening pathway. KSH catalyzes the transformation of 4-androstene-3,17-dione (AD) into 9-OH-AD specifically. In the present study, the putative KshA and KshB genes were cloned from Mycobacterium smegmatis mc(2)155 and Gordonia neofelifaecis NRRL B-59395 respectively, and were inserted into the expression vector pNIT, the co-expression plasmids of kshA-kshB were obtained and electroporated into Mycobacterium sp. NRRL B-3805 cells. The recombinants were used to transform steroids, the main product was characterized as 9α-hydroxy-4-androstene-3,17-dione (9-OH-AD), showing that kshA and kshB were expressed successfully. Different from the original strain Mycobacterium sp. NRRL B-3805 that accumulates 4-androstene-3,17-dione, the recombinants accumulates 9α-hydroxy-4-androstene-3,17-dione as the main product. This results indicates that the putative genes kshA, kshB encode active KshA and KshB, respectively. The process of biotransformation was investigated and the results show that phytosterol is the most suitable substrate for biotransformation, kshA and kshB from M. smegmatis mc(2)155 seemed to exhibit high activity, because the resultant recombinant of them catalyzed the biotransformation of phytosterol to 9-OH-AD in a percent conversion of 90%, which was much higher than that of G. neofelifaecis NRRL B-59395. This study on the manipulation of the ksh genes in Mycobacterium sp. NRRL B-3805 provides a new pathway for producing steroid medicines.
Androstenedione ; analogs & derivatives ; biosynthesis ; Bacterial Proteins ; genetics ; metabolism ; Biotransformation ; Ketosteroids ; Mixed Function Oxygenases ; genetics ; metabolism ; Mycobacterium ; metabolism ; Mycobacterium smegmatis ; enzymology ; Plasmids

To investigate the cellular immunity abnormalities of patients with rheumatoid arthritis(RA), peripheral blood lymphocytes subpopulations, interleukin 2 (IL-2) production and natural killer (NK) cell activity were determined in 9 patients with RA. The results showed that there were a remarkable decrease in NK cell activity and poor response to IL-2 stimlulation. IL-2 production and.the expression of membrane-bound IL-2 receptor were increased in RA patients. The percentage of T4 positive cells and the ratio of T4/T8 were also increased in patients with RA. The results indicate that there are severe cellular immunity abnormalities in patients with RA.

Abnormal activation of Wnt signaling pathway is closely related to the occurrence of tumor, and T cell factor 4 (Tcf4 ) and beta-catenin are important signal transmission factors of this pathway. The aim of the present study is to explore the significance and correlation between expression of Tcf4, beta-catenin and secreted frizzled related protein 1(SFRP1), suppressor gene of Wnt signaling pathway, in colorectal carcinoma and their correlations to the clinicopathological factors. The expressions of Tcf4, beta-catenin and SFRP1 were performed with immunohistochemistry staining in 97 cases of primary colorectal carcinoma and 40 cases of normal colorectal mucosa tissues. The results showed that the abnormal expression rates of Tcf4 and beta-catenin in colorectal carcinoma were significantly higher than those in the control groups (P<0.01). The positive rate of SFRP1 was significantly lower than those in the control groups (P<0.01). The abnormal expression rates of Tcf4 and beta-catenin were also related to the lymph node metastasis and Dukes stage (P<0.05). A significant correlation was found between the expressions of SFRP1 and Tcf4, beta-catenin (P<0.05). Overexpression of Tcf4 and beta-catenin was related to poor prognosis (P<0.05). But the survival rates of the group with SFRP1 expressions were higher than those in group without SFRP1 expressions (P<0.05). Cox multifactor regression analysis indicated that Dukes stage, expression of beta-catenin and SFRP1 were independent risk factors of colorectal carcinoma (P<0.05). The results suggested that the abnormal expression of Tcf4 and beta-catenin in colorectal cancer may be related to the reduced or absent expression of SFRP1. beta-catenin accumulation in the nuclei formed complexes with Tcf4 is one of the important molecular switch maintaining colorectal malignant phenotype. The combined detection of these indexes may perform an important role in predicting the progression and prognosis of colorectal cancer, and could provide new molecular targets for gene treatment of colorectal cancer.
Carcinoma ; metabolism ; Colorectal Neoplasms ; metabolism ; Disease Progression ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Lymphatic Metastasis ; Membrane Proteins ; metabolism ; Phenotype ; Prognosis ; Risk Factors ; Transcription Factor 7-Like 2 Protein ; metabolism ; Wnt Signaling Pathway ; beta Catenin ; metabolism

Objective To observe the effect of 2450 MHz microwave on the white blood cells (WBC) and haematogenous of granulocytic series of mice.Methods The whole body of BALB/c mice were exposure to 2450 MHz power density (10 mW/cm2) microwave, and the mice were killed at different times after exposure, to determine the changes of spleen/body ratio, peripheral blood WBC, number of marrow nucleus cells, cell cycle and form ability of GM-CFU (granule and macrophage-clone forming unit).Results The number of peripheral blood WBC increased at first and then reduced with the exposure time prolonged. Number of marrow nucleus cells kept decreasing after microwave exposure, otherwise, form ability of GM-CFU of marrow cells increased. Exposure to 2450 MHz 10 mV/cm2microwave might speed up marrow nucleus cell passed from G1 period to G2 and S periods.Conclusion Low frequency of 2450 MHz microwave exposure has significant stimulate function on granule cell system, but with the time prolong, number of nucleus cells decreased.

OBJECTIVE: To survey on men who have sex with men (MSM) population's sexual behaviors, condom-service condition, HIV related knowledge and other issues among MSM population at sexually transmitted disease (STD) clinics to understand the characteristics of behaviors and offer suggestions for effective health education and behavior intervention. METHODS: From January to December, 2009, we used anonymous questionnaires which involved in their mastery of demographic characteristics, sexual behavior, condom-service condition, HIV related knowledge, and so one, to investigate 200 MSM at 3 STD clinics of comprehensive hospital. RESULTS: The average age of informant was (26.7+/- 8.9) years and 121 individuals (62.6%) had confirmed with STD in recent one year. In the recent 6 months, the average number of homosexual partners was 9.2+/- 4.8 and 102 (52.8%) had heterosexual partners. In the sexual intercourse with homosexual, 123 individuals had anal intercourse (63.7 %) and 117 had oral intercourse (60.6%). In the sexual intercourse with heterosexual, 92 (90.2%) individuals had vaginal intercourse, 37 (36.2%) had anal intercourse, and 59 (57.8%) had oral sex behavior.There were a statistical difference between heterosexual and homosexual sex behaviors (P<0.01). The condom-using frequency had statistic difference in different sexual behaviors(χ²=188.396, P<0.001). There was no linear correlation between HIV related knowledge and condom-using condition in sexual behaviors except the heterosexual anal intercourse. CONCLUSION High AIDS knowledge mastery rate is found in our survey. The respondents get HIV/AIDS knowledge through various ways actively. There is no obvious relation between the mastery of HIV related knowledge and condom-using frequency. The ratio of non-protected sexual behaviors is high in heterosexuals. How to adopt effective methods for behavior intervention to MSM at STD clinic needs to be further studied.
Acquired Immunodeficiency Syndrome ; prevention & control ; transmission ; Adolescent ; Adult ; China ; epidemiology ; Condoms ; statistics & numerical data ; Female ; Health Knowledge, Attitudes, Practice ; Homosexuality, Male ; Humans ; Incidence ; Male ; Middle Aged ; Safe Sex ; Sexual Behavior ; Sexually Transmitted Diseases ; epidemiology ; Surveys and Questionnaires ; Young Adult

OBJECTIVE: To detect variants of the MMACHC gene among 110 ethnic Han Chinese pedigrees affected with metabolic deficiency methylmalonic acidemia (MMA) of cobalamin C (cblC). METHODS: Peripheral blood samples were collected from the probands and their parents. Following DNA extraction, the coding regions of the MMACHC gene were subjected to PCR amplification, Sanger sequencing and quantitative PCR assaying. For 48 pedigrees, chorionic villus samples were taken for prenatal genetic diagnosis. RESULTS: Thirty five types of variants were detected among the 110 pedigrees, which included missense, nonsense, frameshifting, splicing variants and exonic deletions. Most variants have occurred in exons 4 (73.18%). The detection rate for c.609G>A (p.Trp203Ter) variant was the highest (33.64%), followed by c.658_660delAAG (12.27%), c.567dupT (9.09%) and c.80A>G (6.82%). Two variants, namely c.57_58insT (p.Gly20Trpfs*14) and c.505_506delAT (p.Ile169Argfs*12), were unreported previously and both were of frameshifting types. For the 48 pedigrees undergoing prenatal diagnosis, 14 fetuses were found to be normal, 24 have carried heterozygous variants, the remaining 10 have carried compound heterozygous or homozygous variants. CONCLUSION The discovery of the two novel variants has expanded the spectrum of the MMACHC gene variants among ethnic Han population. Above finding has provide a basis for the prenatal diagnosis and genetic counseling for the affected pedigrees.
Amino Acid Metabolism, Inborn Errors/genetics* ; China ; DNA ; Female ; Humans ; Mutation ; Oxidoreductases/genetics* ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; Vitamin B 12/genetics*