Objective Investigation of biological characteristics of Cdc 20AAA/+APCmin/+ mouse embryonic fibroblast(MEFs) indicate the effect of Cdc20AAA/+on growth of mouse embryonic fibroblast and the possible mechanism . Methods MEFs of Cdc20AAA/+APCmin/+, Cdc20AAA/+, APCmin/+ and WT genotype were harvested from embryos for analysis.The growth characteristics of Cdc20AAA/+APCmin/+, Cdc20AAA/+,APCmin/+and WT mouse embryonic fibroblast were analyzed through growth curve analysis and foci formation assay .Separation of sister chromatid and the presence of aneuploid were detected by karyotype analysis .Results Cell proliferation assays showed that Cdc 20AAA/+APCmin/+cells grew at an accelerated rate compared with APC min/+MEFs(P<0.01).Foci formation assay showed that the clone forming ability was significantly increased .Cdc20AAA/+APCmin/+MEFs showed a significant increase in the frequency of aneuploid compared with WT MEFs , which had a karyotype of 38 and contained prematurely separated sister chromatids .Conclusion Cdc20 carrying a null allele (Cdc20AAA/+) may accelerate the growth and proliferation of APC min/+MEFs and present the growth characteristics of the tumor cells .The possible mechanism may be associated with chromosome instability .

Female ; Humans ; Neoplasm Metastasis ; Uterine Cervical Neoplasms ; pathology

OBJECTIVETo observe the effect of Shenluotong Decoction (SD) on serum levels of aldosterone, monocyte chemoattractant protein-1 (MCP-1), α-smooth muscle protein (α-SMA), and nuclear factor-KB (NF-κB) in obstructive nephropathy rats, and to explore the initial mechanism of SD for inhibiting renal interstitial fibrosis.

METHODSTotally 48 healthy Wistar rats were randomly divided into the sham-operation group (n =12) and the model group (n =36). Renal interstitial fibrosis rat model was established by unilateral ureteral obstruction (UUO). After successful modeling, 36 rats were randomly divided into the model group, the Chinese medicine group, and the Western medicine group, 12 in each group. Eplerenone was added in the forage at the daily dose of 100 mg/kg for rats in the Western medicine group. Chinese medicine was added in the forage at the daily dose of 26 g/kg for rats in the Chinese medicine group. Equal volume of normal saline was administered to rats in the sham-operation group and the model group. All medication was performed once daily. The obstructive kidneys were extracted ten days after medication. The pathomorphological changes were observed. The contents of serum aldosterone and MCP-1, and the protein or mRNA expression of MCP-1, α-SMA, and NF-KB were detected.

RESULTSCompared with the sham-operation group, infiltration of a large amount of inflammatory cells and collagen deposition significantly increased, serum contents of aldosterone and MCP-1 obviously increased (P < 0.01), the expression of MCP-1 mRNA and protein were significantly up-regulated (P <0.01), the protein expression of α-SMA and NF-KB were significantly enhanced in the model group (P <0.01). Com- pared with the model group, infiltration of inflammatory cells and renal collagen deposition were attenua- ted in the Chinese medicine group and the Western medicine group, the serum MCP-1 level were reduced, and the mRNA and protein expression of MCP-1 were significantly down-regulated (P <0.01), the protein expression of α-SMA and NF-KB were obviously inhibited (P <0. 01). At the same time, serum aldosterone level was reduced in the Chinese medicine group (P <0.01).

CONCLUSIONSinflammatory lesions of the renal tissue could promote the progress of interstitial fibrosis in rats with obstructive nephropathy. SD could attenuate interstitial fibrosis through reducing serum contents of aldosterone and MCP-1, down-regulating MCP-1/ NF-KB, and inhibiting the expression of α-SMA.

Animals ; Chemokine CCL2 ; drug effects ; metabolism ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fibrosis ; Kidney ; drug effects ; Kidney Diseases ; drug therapy ; genetics ; NF-kappa B ; drug effects ; metabolism ; RNA, Messenger ; biosynthesis ; Rats, Sprague-Dawley ; Ureteral Obstruction ; drug therapy ; genetics

OBJECTIVETo evaluate the clinical features of patients with primary biliary cirrhosis (PBC) and positive expression of sp100 autoantibody in order to generate a clinical screening profile that may help to increase early diagnosis and timely initiation of therapy.

METHODSThe clinical data of 70 patients who were diagnosed with PBC by liver biopsy between January 2006 to December 2009 at the Second Affiliated Hospital of Kunming Medical University of Hepatobiliary and Pancreatic Medicine were retrospectively collected for analysis. The patients were divided according to expression of anti-sp100: positive patients, n = 12; negative patients, n = 58. The groups were comparatively analyzed for differences in clinical, biochemical, immunological, and histopathological parameters. Normally distributed data was compared by t-test, and non-normally data was compared by rank-sum test.

RESULTSThere was no significant difference in age among the sp100-positive and sp100-negative patients (51.6 +/- 9.5 vs. 50.0 +/- 14.7 years, P more than 0.05). The sp100-positive group had significantly more women (80.0% vs. 61.9%, X2 = 0.32, P more than 0.05) and more patients with atypical symptoms (18.2% vs. 13.8%) but the difference of the latter did not reach statistical significance. The sp100-positive group had significantly higher levels of alkaline phosphatase (ALP; 466 vs. 163 U/L, Z = 3.71), gamma-glutamyl-transpeptidase (GGT; 728 vs. 154 U/L, Z = 3.38), and immunoglobulin M (IgM; 4.25 +/- 2.86 vs. 2.81 +/- 2.15, t = 2.06, P less than 0.05). Forty of the total patients tested negative for antimitochondrial (AMA)-M2 antibodies, and eight of those were sp100-positive (20.0%) while 18 were antinuclear (ANA) antibody-positive (45.0%). There were significantly more AMA-M2-negative/ANA-positive patients than sp100-positive patients (P = 0.021). Anti-sp100 expression was not associated with the pathological stage of PBC (R1 = 5.500, P more than 0.05).

CONCLUSIONSP100-positive PBC may show a bias towards the female sex, and may be characterized by enhanced serum levels of ALP, GGT, and IgM. Further clinical differences may manifest as the disease progresses, and changes in autoantibodies' expression and liver function markers should be carefully monitored in follow-up.

Adult ; Aged ; Antibodies, Antinuclear ; blood ; Antigens, Nuclear ; immunology ; Autoantibodies ; blood ; Autoantigens ; immunology ; Female ; Humans ; Liver ; pathology ; Liver Cirrhosis, Biliary ; immunology ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

OBJECTIVETo evaluate the influence of Chinese drugs for cooling blood and dissolving stasis (CBDS) in different concentrations on morphology of krypton laser induced choroidal neovascularization (CNV) in brown Norway (BN) rats.

METHODSForty-eight rats received laser irradiation (659 nm) on fundus of one eye (power 360 mW, spot diameter 50 microm, time 0.05 s). They were divided into four groups equally: the control group (A) treated with normal saline, and the three CBDS groups treated respectively with high (B, 5.0 g/kg), median (C, 2.5 g/kg) and low (D, 1.25 g/kg) dosage of CBDS, twice every day via gastric perfusion for 21 successive days. Fundus fluorescein angiography (FFA) on 4 selected rats in each group was performed at the 7th, 14th and 21st day after photocoagulation, and histopathologic examination using light microscope with immuno-histochemical stain was conducted on them as well.

RESULTSFFA showed that CNV was firstly appeared on day 7 after photocoagulation, in Group A, it expanded gradually and reached the peak on day 21, but showed no significant expansion in the three CBDS groups. The fluorescein leakage in Group C (52343.13 +/- 12973.92 dots) and D (66252.78 +/- 20659.71 dots) was significantly less than that in Group A (91457.19 +/- 29309.11 dots) and B (95973.40 +/- 53950.43 dots) on day 21, all showing statistical significance (P<0.05). The variation of CNV in thickness showed that in Group A it increased gradually from day 7 and reached the peak on day 21 (55.3383 +/- 8.5036 microm); but in the CBDS groups, the peak was reached on day 14, then became thinned gradually, on day 21, it was 40.0913 +/- 13.3448 microm in Group B, 38.8473 +/- 7.9862 microm in Group C and 38.9372 +/- 5.1728 microm in Group D, all thinner than that in Group A significantly (P<0.05).

CONCLUSIONCDBS can effectively suppress the krypton laser induced CNV proliferation and prevent the CNV leakage in BN rats.

Animals ; Choroidal Neovascularization ; drug therapy ; pathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Fluorescein Angiography ; Male ; Rats ; Rats, Inbred BN

OBJECTIVETo evaluate the effects of microscopic observation drug susceptibility (MODS) in detecting susceptibility of Mycobacterium tuberculosis (MTB) onto four first line anti-tuberculosis drugs.

METHODThe 24-hole cell culture plates were used to test drug susceptibility of MTB on liquid medium, and the best detecting condition of MODS assay was probed; 66 clinical isolates susceptibility to streptomycin (S), isoniazid (H), rifampin (R) and ethambutal (E) were evaluated by using MODS assay and Lowenstein-Jensen (L-J), thereafter, all the inconcordance of isolates between MODS and L-J were tested for the minimal inhibitory concentrations (MIC).

RESULTSConcordance rate of the susceptibility to S, H, R and E in 66 clinical isolates detected by MODS and L-J was 97.0%, 90.9%, 95.5% and 86.4% respectively. If the results obtained by L-J were taken as a golden standard, the sensitivity, specificity, positive and negative predictive value (PPV and NPV) as well as accuracy of susceptibility test to S detected by MODS was 96.0%, 97.6%, 96.0%, 97.6% and 97.0%; 100%, 85.4%, 81.0%, 100% and 90.9% to H; 96.2%, 95%, 92.6%, 97.4% and 95.5% to R; 73.7%, 91.5%, 77.8%, 89.6% and 86.4% to E. There were 20 inconsistent results of 16 isolates by comparing MODS with L-J, and MIC yielded 16 results of those 14 isolates showing identical results with those of the MODS, while 4 results of other 4 isolates identical with L-J.

CONCLUSIONMODS method simultaneously provides drug susceptibility to S, H, R and E. MODS might be one of the rapid tools to diagnosing multidrug-resistant tuberculosis as it is rapid, simple, inexpensive and has high concordance with L-J drug susceptibility test.

Bacteriological Techniques ; methods ; Drug Resistance, Multiple, Bacterial ; Microbial Sensitivity Tests ; methods ; Mycobacterium tuberculosis ; drug effects ; Predictive Value of Tests ; Sensitivity and Specificity ; Tuberculosis, Multidrug-Resistant ; microbiology

OBJECTIVETo explore the effects of cofilin on the actin cytoskeleton reorganization in osteoblasts induced by fluid shear stress.

METHODSFluid shear stress (1.2 Pa) was applied to osteoblasts for 0 (control group), 15, 30, 45, 60, 120 min in vitro. Cells were stained with fluorescein isothiocyanate (FITC)-phalloidin for fiber-actin, and confocal laser scanning microscope(CLSM) was used to observe the fluorescence of fiber-actin. Western blotting was used to detect the expression of the cofilin and the phospho-cofilin.

RESULTSActin filaments became organized into stress fibers that were thicker and more abundant than those in non-flowed cells. The fluorescence intensity (38.00 ± 6.88) of fiber-actin after 120 min (42.93 ± 6.41) loading it was 2.8 times as much as that in control group (15.41 ± 3.60, P < 0.05). Additionally, the level of phospho-cofilin protein was dramatically elevated after loading. Fluid shear stress induced an initial decrease of cofilin at 60 min. However, at 120 min cofilin (0.254 ± 0.026) increased to 1.5 times as much as that at 60 min (0.162 ± 0.004).

CONCLUSIONSThe results indicate that cofilin phosphorylation mediates fiber-actin reorganization in the osteoblasts induced by fluid shear stress.

Actin Cytoskeleton ; ultrastructure ; Actin Depolymerizing Factors ; biosynthesis ; Humans ; Osteoblasts ; ultrastructure ; Phosphorylation ; Stress, Mechanical

OBJECTIVETo investigate the clinical features of Crohn disease according to the Montreal classification.

METHODSClinical data of 43 surgical patients with Crohn disease (surgical group) and 125 non-surgical patients with Crohn disease (non-surgical group) were retrospectively analyzed and compared between two groups. The Montreal classification was used.

RESULTSIn the surgical group, 28 patients (65.1%) were A2, 14 (32.6%) were A3 and only one was A1, which was not significantly different as compared to the non-surgery group. The proportions of L1, L2, L3, and L4 subtype in the surgical group were 41.9%, 25.6%, 30.2%, and 2.3%, respectively, which was not significantly different as compared to that in the non-surgery group. In the surgical group,B1 disease was found in 1 case (2.3%), B2 in 26 cases (60.5%), and B3 in 16 cases (37.2%), while in the non-surgical group, B1 was found in 79 cases (63.2%), B2 in 44 cases (35.2%) and B3 in 2 cases (1.6%). Differences were significant between two groups in disease behavior (P=0.001, P=0.004, P=0.001).

CONCLUSIONSMost surgical patients of Crohn disease are A2. L1 and L3 are the main lesion location. As disease behavior, B2 and B3 are the main reasons for operation.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Crohn Disease ; classification ; pathology ; surgery ; Female ; Humans ; Male ; Middle Aged ; Reference Standards ; Retrospective Studies ; Young Adult

Animals ; Disease Models, Animal ; Fatty Liver ; Male ; Mice ; Mice, Inbred C57BL

Twin pregnancy with mosaic partial hydatidiform mole (PHM) and survival of two healthy fetuses following in vitro fertilization and embryos transfer (IVF-ET) is a rare situation and is considered a challenge for management. A 32-year-old Chinese woman conceived twin pregnancy following IVF-ET. At 22 weeks' gestation, an additional intrauterine echogenic mass with features of PHM were shown by successive ultrasound examinations. At 35 weeks' gestation, two live male infants and two placentas were delivered by caesarean section (CS). Histologic examination of the abnormal placenta confirmed mosaic PHM. Genetic study showed the abnormal placental mosaicism (expressed in molar-69XXY and normal vili-46XY), co-existing with a hypospadia new-born (46XY) in one amniotic sac. However, the other one was normal. Serial serum β-hCG levels showed a declining trend and serum β-human chorionic gonadotropin (hCG) were undetectable at 6 months after delivery. The case demonstrated that it is possible to prolonged gestation by PHM under close surveillance during the entire pregnancy.
Adult ; Female ; Fertilization in Vitro ; Humans ; Hydatidiform Mole ; genetics ; Infant, Newborn ; Male ; Mosaicism ; Placenta ; metabolism ; pathology ; Pregnancy ; Pregnancy, Twin ; genetics