Breast Neoplasms ; chemistry ; Female ; Humans ; Immunohistochemistry ; methods ; standards ; In Situ Hybridization, Fluorescence ; methods ; standards ; Receptor, ErbB-2 ; analysis ; Stomach Neoplasms ; chemistry

OBJECTIVETo observe the synergistic effect of Qilian decoction (QLD) with corresponding hypoglycemic agents on insulin sensitivity in patients with diabetes mellitus type 2 and its influence on related inflammatory cytokines.

METHODSSixty-two patients with diabetes mellitus type 2 were selected and randomly divided into two groups, they were treated with hypoglycemic agent in routine, and QLD was given orally, one dose taken in twice a day. Parameters as fasting blood glucose, insulin, peptide C, tumor necrosis factor-alpha (TNF-alpha), C-reactive protein (CRP), interleukin 6 (IL-6) were measured before and after treatment, and insulin sensitive index (ISI) was also calculated.

RESULTSThe level of fasting blood glucose lowered after treatment in both groups (P<0.01); levels of fasting insulin, peptide C, ISI, TNF-alpha, IL-6 and CRP significantly lowered after treatment (P<0.05 or P<0.01), and the effect was better than that in the control group (P<0.05 or P<0.01).

CONCLUSIONQLD could improve the insulin resistance and lower the levels of related inflammatory cytokines.

Aged ; C-Reactive Protein ; metabolism ; Diabetes Mellitus, Type 2 ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hypoglycemic Agents ; therapeutic use ; Insulin ; blood ; therapeutic use ; Insulin Resistance ; Interleukin-6 ; blood ; Male ; Middle Aged ; Peptides ; blood ; Phytotherapy ; Tumor Necrosis Factor-alpha ; metabolism

The aim of this study was to investigate the effects of whole-body vibration on Wnt/β-catenin signaling pathway in bone marrow cells of ovariectomized osteoporosis rats. Thirty-six healthy 3-month old female Sprague Dawley (SD) rats were randomly divided into the following three groups by body weight: sham-operation (Sham), ovariectomized (OVX), and OVX whole-body vibration (WBV) groups. Ten weeks after ovariectomization, the rats of WBV group received vibration treatment (90 Hz, 15 min) twice per day. At the end of 8-week vibration, the whole-body bone mineral density (BMD) and body composition were detected by dual energy X-ray absorptiometry (DEXA) in vivo. The protein expressions of β-catenin and p-GSK3β in both bone marrow cells and bone marrow stromal cells were detected by Western blot. The results showed that, compared with OVX group, WBV group showed decreased fat mass and fat mass content, as well as increased lean body mass content. The BMD of the proximal tibia in WBV group was significantly higher than that in OVX group, however, there was no difference of BMD in whole-body and other positions between the two groups. The β-catenin expression in bone marrow stromal cells showed no difference between OVX and WBV groups. The p-GSK3β expression of bone marrow cells was increased in WBV group compared with that in OVX group, whereas bone marrow stromal cells from two groups did not exhibit the difference of the p-GSK3β expression. These results suggest that whole body vibration can stimulate the protein expression of p-GSK3β in bone marrow cells of ovariectomized osteoporosis rats, which could improve the bone loss induced by ovariectomization.
Absorptiometry, Photon ; Animals ; Body Composition ; Body Weight ; Bone Density ; Bone Marrow Cells ; metabolism ; Disease Models, Animal ; Female ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Osteoporosis ; metabolism ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Tibia ; Vibration

OBJECTIVETo identify the ETV6 gene rearrangement in patients with myelodysplastic syndromes (MDS) and explore its relationship with prognosis and disease stages.

METHODSETV6 rearrangement in 58 MDS cases were detected by conventional cytogenetics and Split-signal FISH. RT-PCR was used to detect 9p24-12p13 balance translocation with special designed primers ETV6F1/F2 and JAK2R1/R2. The relationship between ETV6 rearrangement and prognosis and disease staging in MDS patients was analyzed.

RESULTSETV6 rearrangement were found in 4 (6.9%) of 58 cases, among which ETV6/JAK2 fusion was identified by RT-PCR in 1 (1.7%) case. The mean follow-up duration was 12 months. All 4 patients (100%) with rearrangement transformed into acute leukemia, with a median survival time (MS) of 7 months; while 10 patients (17%) in the non-translocation group transformed to acute leukemia, with a MS of 28 months. In addition, all 4 patients (100%) with rearrangement were in advanced stage of MDS( RAEB), while 17 cases (31.5%) in non-rearrangement group were in that stage.

CONCLUSIONSETV6 rearrangement has higher expression rate (6.9%), and is closely associated with disease stage and prognosis in MDS.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Gene Rearrangement ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; pathology ; Neoplasm Staging ; Prognosis ; Proto-Oncogene Proteins c-ets ; genetics ; Repressor Proteins ; genetics

OBJECTIVETo investigate the feasibility of real-time fluorescent quantitative (qPCR) assay in detecting mycobacterium tuberculosis complex (MTB) in paraffin embedded tissues for diagnostic purpose.

METHODSUsing qPCR assay, 1000 consecutive formalin-fixed and paraffin embedded (FFPE) tissues (from 2011 to 2012) suspected of MTB infection were tested by amplifying the MTB specific insertion sequence 6110 (IS6110). The specificity of the PCR product was confirmed by Sanger sequencing as compared with the MTB genomic DNA of the IS6110 sequence. Tissues with Ziehl-Neelsen acid-fast staining were used as control.

RESULTSIn the 1000 samples, 513 were positive for mycobacterium by Ziehl-Neelsen acid-fast staining (detection rate 51.3%); whereas 546 were MTB positive by qPCR assay (detection rate 54.6%). Concordance rate for both assays was 73.1%. The diagnosis rate increased by 14.4% by combinination of Ziehl-Neelsen acid-fast staining and qPCR results. More interestingly, by analyzing the Ziehl-Neelsen acid-fast staining and qPCR results three cases of M.leprae infection and four cases of non-tuberculous Mycobacterium (NTM) infection were identified.

CONCLUSIONSqPCR detection of MTB in FFPE tissue is more sensitive than Ziehl-Neelsen acid-fast staining assay. Combination of these two assays can increase the detection rate and also identify some rare cases of NTM infection.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; DNA, Bacterial ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mycobacterium tuberculosis ; genetics ; isolation & purification ; Paraffin Embedding ; Real-Time Polymerase Chain Reaction ; Sequence Analysis, DNA ; Staining and Labeling ; methods ; Tuberculosis ; diagnosis ; microbiology ; Tuberculosis, Gastrointestinal ; diagnosis ; microbiology ; Tuberculosis, Lymph Node ; diagnosis ; microbiology ; Tuberculosis, Pulmonary ; diagnosis ; microbiology ; Young Adult

OBJECTIVETo investigate the advantages and disadvantages of dual-color silver-enhanced in-situ hybridization (DSISH) and fluorescence in-situ hybridization (FISH) for determination of HER2 gene status in gastric carcinoma and to evaluate the feasibility of DSISH.

METHODSEighty cases of primary gastric or gastroesophageal junction adenocarcinomas diagnosed and treated surgically from January to March, 2009 at the West China Hospital were enrolled in the study. Automated immunohistochemistry (IHC) staining, FISH and automated DSISH were carried out to detect the HER2 status, respectively, and the concordance of the three techniques was then evaluated.

RESULTSDSISH and FISH failed initially, but repeated detection was successful in 5 cases. Gene amplification was detected in 12/13 IHC 3+ cases in DSISH and in 11/13 IHC 3+ cases in FISH. In 6 IHC 2+ cases, the amplification rate was both 1/6; in 18 IHC 1+ cases, the amplification rate was both 2/18. No amplification was observed in 43 IHC 0 cases. Only one of the 80 cases showed discrepancy, and therefore the overall concordance between FISH and DSISH was 98.8% (κ = 0.958, P < 0.01).

CONCLUSIONSDSISH represents a novel approach for the determination of HER2 status in gastric carcinoma, and the overall concordance between DSISH and FISH is excellent. Despite their advantages and disadvantages, DSISH is more feasible and practical for routine application in surgical pathology.

Adenocarcinoma ; genetics ; Esophagogastric Junction ; Gene Amplification ; Genes, erbB-2 ; Humans ; In Situ Hybridization ; methods ; In Situ Hybridization, Fluorescence ; Sensitivity and Specificity ; Silver ; Stomach Neoplasms ; genetics

OBJECTIVETo observe and compare the luciferase activities of different length segments of human dentin matrix protein 1 promoter in human dental pulp stem cells (HDPSC), osteoblasts (OC) and Hela cells.

METHODSThe differentlength desired DNA segments were obtained from 2 195 bp Dmp1 promoter cloned by PCR method. The amplified promoter segments with different length were cloned into luciferase report gene vector pGL3-Basic, the correct orientation of those inserts was verified by cutting with two different restrict enzymes. The luciferase activity was observed after different pGL3-PDmp1 vectors were transfected transiently into those three different-type cells.

RESULTS6 Dmp1 promoter segments with different-length were obtained successfully, and luciferase report gene vectors with different promoter segments were successfully constructed after identified by restriction enzymes cutting. They had different luciferase activities when they were transfected transiently into HDPSC, and the region of -505(-)-193 bp and -935(-)-505 bp could be regarded as the specific promoters of Dmp1 promoter for HDPSC and OC respectively, which could include the basic regulatory elements.

CONCLUSIONThe correct clone of the upstream of human Dmp1 promoter segments with different length had been obtained, and they had strong luciferase activities in HDPSC and OC, but very low in Hela cell. These results will make an important basis for studying mineralized tissue-specific transcriptional regulation mechanisms of Dmp1.

Dentin ; Extracellular Matrix Proteins ; Gene Expression Regulation ; Genetic Vectors ; Humans ; Phosphoproteins ; Promoter Regions, Genetic ; Transfection

OBJECTIVETo investigate the mechanism of all trans retinoid acid (ATRA) inhibition of cell growth and induction of apoptosis in human retinoblastoma Y79 cells.

METHODSAntiproliferating effects of ATRA on Y79 cells were studied by (3)H-thymidine incorporation. Cell cycle analysis was performed by flow cytometry, apoptosis of the ATRA-treated cells was determined by DNA fragmentation analysis and JNK phosphorylation analyzed by Western blot.

RESULTSAfter 36h treatment of 1 micro mol/L ATRA, (3)H-thymidine incorporation decreased to 40% with Y79 cells arrested in G(0)/G(1) and Sub-G(1) peak appeared. DNA ladder was observed in DNA fragmentation analysis after 36h treatment of ATRA. Curcumin, a JNK blocker, blocked the apoptosis and the growth inhibition induced by ATRA. JNK was phosphorylated in 10 to 20 min.

CONCLUSIONATRA can induce the apoptosis in Y79 cells by phosphorylation of JNK, which suggests that ATRA may have clinical application prospects for treatment of retinoblastoma.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Flow Cytometry ; Humans ; JNK Mitogen-Activated Protein Kinases ; physiology ; Phosphorylation ; Retinoblastoma ; drug therapy ; pathology ; Thymidine ; metabolism ; Tretinoin ; pharmacology

OBJECTIVETo evaluate the clinical value of HER2 overexpression in breast cancer and its prognostic implication in patients with lymph node negative breast carcinoma.

METHODSThe following electronic database were extracted using appropriate inclusive and exclusive standards: Cochrane library, PUBMED, Embase (1984 - 2003), OVID, CMCC and CNKI. Excel and RevMan 4.2 were used for statistical analysis.

RESULTSFifty-six articles were extracted to calculate the positive rate of HER2 overexpression. The pooled positive rate was 23.14% [19.54%, 26.73%], with positive immunohistochemistry (IHC) rate of 23.13% [19.49%, 26.77%] and positive FISH rate of 20.90% [15.54%, 26.25%]. Seven articles were used to evaluate prognostic predication of HER2 expression. It was concluded that in patients with lymph node negative breast carcinoma, HER2 overexpression (both IHC and FISH) independently predicted a poor prognosis based on disease-free survival (DFS) and overall survival (OS) with a P < 0.05. For DFS, the pooled RR was 1.38 [1.07, 1.80] with 1.16 [1.02, 1.31] for IHC and 1.98 [1.56, 2.52] for FISH. For OS, the pooled RR was 1.58 [1.16, 2.14] with 1.37 [1.14 to 1.64] for IHC and 2.33 [1.45 to 3.75] for FISH. HER2 overexpression effectively predicted DFS/OS of patients without adjuvant therapy and OS of patients with the therapy, but not for DFS, with the pooled RR of 1.46 [1.02, 2.09] and 1.11 [0.95, 1.31] for DFS, respectively and the pooled RR of 1.93 [1.44 to 2.58] and 1.25 [1.01, 1.56] for OS, respectively.

CONCLUSIONSIn patients with lymph node negative breast carcinoma, the positive rate of HER2 overexpression is 23.14%. HER2 overexpression indicates a poor prognosis and adjuvant therapy after surgery should be recommended.

Breast Neoplasms ; genetics ; metabolism ; pathology ; therapy ; Chemotherapy, Adjuvant ; Disease-Free Survival ; Female ; Genes, erbB-2 ; Humans ; Lymph Nodes ; pathology ; Mastectomy ; Prognosis ; Receptor, ErbB-2 ; metabolism ; Survival Rate

OBJECTIVETo observe the diagnostic value of non-invasive 128-slice computed tomography coronary angiography (CTA) in comparison with invasive coronary angiography.

METHODS128-slice CTA and invasive coronary angiography were performed in 78 unselected consecutive patients (63 patients with suspected coronary artery disease and 15 patients with previous coronary stenting, 56 males, mean age 61 +/- 10 years) and > 50% reduction of minimal lumen diameter was defined as significant coronary stenosis.

RESULTSFifty-eight out of 879 segments (7%) from CTA were not assessable because of irregular rhythm, vessel calcification or tachycardia. Compared with invasive coronary angiography, segment-based analysis from the 821 segments showed the sensitivity by CTA was 87%, specificity 97%, PPV 83% and NPV 97%. Four out of 22 stents implanted in 15 patients were not assessable by CTA because of poor image quality. Compared with invasive coronary angiography, the sensitivity of diagnosing in-stent restenosis by CTA was 100%, specificity 77%, PPV 63% and NPV 100% for the remaining 18 stents.

CONCLUSIONSOne hundred and twenty-eight-slice CTA has a high accuracy for detecting coronary artery disease and in-stent restenosis after coronary stenting and could be considered as a valuable noninvasive technique for screening coronary artery disease in suspected patients.

Adult ; Aged ; Coronary Angiography ; methods ; Coronary Artery Disease ; diagnosis ; diagnostic imaging ; Coronary Stenosis ; diagnosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Tomography, Spiral Computed ; methods