AIM: To observe the effect of Shenmai injection,a chinese medicine,on apoptosis of cardiac myocytes after hypoxia.METHODS: Cardiac myocytes were separated from neonate rat heart and cultivated in vitro.Hypoxia condition was induced by mixture of 95%N2 and 5%CO2.Cells were exposed to hypoxia for 6 h or 12 h and treated with Shenmai injection(5 mL/L) from 24 h before hypoxia until the end of hypoxia.First,apoptosis was detected with Annexin V-FITC and PI staining by flowcytometry.Then,the activity of cardiac myocyte mitochondria was observed by MTT method.Mitochondria membrane potential and the activity of caspase 3,7 were also measured by laser scan microscopy and multi-detection microplate reader,respectively.RESULTS: The apoptotic cells became more and more with prolonged hypoxia.Shenmai injection enhanced mitochondria activity,kept membrane potential,inhibited the activation of caspase3,7 and then decreased apoptotic cells(P

OBJECTIVETo study the effect of Xinnao Shutong capsule (XNST) on energy metabolism dysfunction, free radical injury and inflammatic factors in the course of acute cerebral ischemic damage, and try to reveal the mechanism of the protection against ischemia.

METHOD60 male Wistar rats weighing 280 - 320 g were randomly divided into five groups: normal, sham operation, model, XNST treatment( XNST-T) , and Western medicine treatment (WM-T) group. Acute multi-infarct model in rats was induced by injecting the embolus of blood powder through the right external carotid artery (ECA) into the internal carotid artery (ICA). At 72 hours after ischemia, morphologic change and the express of tumor necrosis factor-alpha (TNF-alpha) and interleukin -1beta ( IL-1beta) in hippocampus CAl section and cortex were observed, biochemical criterions including the activity of Na+ -K+ -ATPase, lactate dehydrogenase (LDH), superoxide dismutase (SOD), and the content of malondialdehyde (MDA) in hippocampus were examined.

RESULTThe morphologic change of hippocampus and cortex in both XNST-T and WM-T groups was milder than that in model group. The activity of Na+ -K+ -ATPase, LDH and SOD in hippocampus were all significantly decreased in model group (P <0. 01), and elevated in XNST group (P <0. 01) as well as in WM-T group (P <0. 01). The content of MDA in hippocampus was significantly increased in model group (P <0. 05), and was reduced in XNST group (P <0. 05) as well as in WM-T group (P <0. 01).

CONCLUSIONThe results reveal that XNST has the protective effect against cerebral ischemic injury. And its possible mechanism is that XNST can prevent the upper pathological process.

Animals ; Brain Infarction ; complications ; Brain Ischemia ; etiology ; metabolism ; prevention & control ; Capsules ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hippocampus ; drug effects ; metabolism ; pathology ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Saponins ; isolation & purification ; pharmacology ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Superoxide Dismutase ; metabolism ; Tribulus ; chemistry

OBJECTIVETo explore the role of survivin and PI3K/AKT pathway in the pathogenesis of psoriasis vulgaris (PV).

METHODSPlaque-like lesions collected from 22 patients with PV in progressive stage and 18 normal control skin specimens were examined using immunohistochemical staining, Western blotting and real-time quantitative PCR for expressions of survivin, PI3K and AKT in the keratinocytes, and their correlation was analyzed. A small interfering RNA (siRNA) was used to knock down AKT in cultured HaCaT cells, and Western blotting was used to detect the changes in the expression of survivin. RESULTS Compared with normal skin, PV lesions showed obviously up-regulated expressions of survivin, PI3K and AKT in the keratinocytes. Survivin expression was positively correlated with PI3K (r=0.4510, P=0.0351) and AKT (r=0.4423, P=0.0393) in the keratinocytes in PV lesions. In cultured HaCaT cells, siRNA-mediated knockdown of AKT caused down-regulation of survivin expression.

CONCLUSIONSurvivin and PI3K/AKT signaling pathway may participate in the occurrence and progression of PV.

Objective To analyze the clinical performance of free prostate-specific antigen (fPSA)detection by ECLIA method,and evaluate whether ECLIA is suitable for clinical use.Methods 341samples were collected and tested prostate-specific antibodies with CMIA and ECLIA methods.These samples contain:97 samples with abnormal high PSA value tested by CMIA method,and 244 normal PSA samples.Use CMIA as the reference method,and detect fPSA,tPSA levels,and the ratio of fPSA/tPSA.Analyze the testing results with statistical methods.Results Compared with CMIA,correlation coefficent of ECLIA fPSA detection is 0.99; correlation coefficent of f/tPSA ratio detection is 0.96; the sensitivity,specificity of ECLIA f/tPSA ratio detection are 85.71％,92.6％ respectively,the agreement rate with ECLIA is 87.4％.No cross reaction with bilirubin,lipohemia,hemolysis,RF,CEA,AFP,CA125,CA153,CA199 were found in the tests.Conclusion The ECLIA method for free prostate-specific antigen detection showed good clinical performance; and is suitable for clinical use.

OBJECTIVETo identify genetic abnormalities in primary gastric carcinoma.

METHODSComparative genomic hybridization (CGH) was used in screening DNA copy number changes along all chromosomes in 23 cases of primary gastric cancer.

RESULTSTwenty-one out of 23 cases showed chromosomal losses and gains for at least one of the chromosomal arms in primary gastric cancer. The mean number of chromosomal alterations was 7.52. Chromosomal gains predominated over chromosomal losses in a ratio of 5.38:2.14. The most often involved chromosomal gains were observed in 8q (9/21, 42.9%), 20q (9/21, 42.9%), 17q (8/21, 38.1%), 3q (7/21, 33.3%), 7q (7/21, 33.3%), 11q (6/21, 28.6%), 13q (6/21, 28.6%), 1q (5/21, 23.8%) and 20p (5/21, 23.8%). The chromosomal arms with frequent losses were 17p (7/21, 33.3%), 18q (6/21, 28.6%), 5q (5/21, 23.8%), 8p (5/21, 23.8%), and 9p (5/21, 23.8%).

CONCLUSIONSThe phenomenon of gain and loss of chromosomal regions is observed in primary gastric cancer, which may induce the amplification of oncogenes and the loss of tumor suppressor genes to regulate the development and progression of gastric cancer.

Adult ; Aged ; Aged, 80 and over ; Chromosome Aberrations ; Chromosome Deletion ; Comparative Genomic Hybridization ; DNA ; Female ; Gene Expression Profiling ; Genes, Tumor Suppressor ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Neoplasm Staging ; Stomach Neoplasms ; genetics ; pathology

AIMTo identify a novel alternative transcript of the novel retinal pigment epithelial cell gene (NORPEG) expressed in the human testis.

METHODSA human testis cDNA microarray was established and hybridized with cDNA probes from human fetal testes, adult testes and human spermatozoa. Differentially expressed clones were sequenced and analyzed. One of these clones was a short transcript of NORPEG which we proceeded to analyze by RT-PCR.

RESULTSThe novel short alternative transcript of NORPEG was isolated and named sNORPEG. It was 3486 bp in length and contained a 2952-bp open reading frame, encoding a 110.4-kDa protein of 983 amino acids. Amino acid sequence analysis showed that the sNORPEG protein contains six ankyrin repeats and two coiled-coil domains. It shares a high homology with the NORPEG and ankycorbin proteins in both its sequence and motifs. Blasting the human genome database localized sNORPEG to human chromosome 5p13.2-13.3. Expression profiles showed that sNORPEG was expressed in human fetal testes, adult testes and spermatozoa. Moreover, sNORPEG was found to be ubiquitously expressed in human tissues.

CONCLUSIONsNORPEG is expressed in different developmental stages of the testis and encodes a protein that may have roles in human testis development and spermatogenesis.

Alternative Splicing ; Amino Acid Sequence ; Base Sequence ; Cytoskeletal Proteins ; genetics ; DNA, Complementary ; Gene Expression Profiling ; Humans ; Male ; Molecular Sequence Data ; Open Reading Frames ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Amino Acid ; Testis ; metabolism ; Transcription Factors ; genetics

OBJECTIVETo investigate the value of transabdominal ultrasonography (TAUS) in preoperative assessment of TNM stage and tumor angiogenesis for patients with gastric carcinoma.

METHODSSixty- four patients with gastric carcinoma preoperatively underwent TAUS, in whom transabdominal color Doppler ultrasonography was used for measuring color Doppler vascularity index (CDVI) of each tumor in 37 cases and microvessel density (MVD) was evaluated by using immunohistochemical staining of surgical specimens with anti- CD34 antibody.

RESULTSThe overall accuracy rate was 56.0% for T staging of gastric carcinoma (T (1) 2/3 cases, T (2) 28.6% , T (3) 73.1% , T (4) 50.0% , respectively) by TAUS. The diagnostic accuracy rate was 63.3% for lymph node status of gastric carcinoma. The diagnostic sensitivity and specificity for lymph node metastasis was 37.9% and 100% respectively. The overall accuracy for N staging of gastric carcinoma was 57.1% (N (0) 100% , N (1) 16.7% , N (2) 35.3% , respectively). The diagnostic sensitivity and specificity for determining distant metastases was 58.3% and 100% respectively. The CDVI of gastric carcinoma determined by color Doppler ultrasonography was significantly correlated to vascular invasion (P=0.0418), a linear correlation between CDVI and MVD was determined by logistic regression analysis (r=0.5628, P< 0.01).

CONCLUSIONTAUS can be a routine diagnostic approach for preoperative gastric carcinoma patients.

Abdomen ; diagnostic imaging ; Adult ; Aged ; Female ; Humans ; Male ; Microvessels ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; diagnostic imaging ; Stomach Neoplasms ; diagnostic imaging ; pathology ; Ultrasonography, Doppler, Color ; methods

OBJECTIVETo observe the hepatic injury following stop- flow chemotherapy and investigate the potential mechanisms.

METHODSTwelve healthy hybrid female pigs were randomly divided into two groups as stop- flow group (SF) and stop- flow chemotherapy (SFC) group. The expression of IL- 8 and ICAM- 1 mRNA in hepatic biopsies was detected by RT- PCR, and the expression of NF- kappa B P65 subunit in nuclei was assessed by Western blot analysis. The levels of ALT and AST, and histopathologic alterations were examined to evaluate the hepatic function at different time before and after stop- flow procedure.

RESULTSThe expression of NF- kappa B P65 subunit, IL- 8 and ICAM- 1mRNA increased at 30 min after stop- flow procedure, and gradually decreased at 3 h and 6 h after stop- flow procedure. The levels of ALT and AST decreased after reaching the peak at 24 h after stop- flow procedure, but removed one week after stop- flow procedure. Cytoplasmic microvascular steatosis developed with appreciable neutrophils infiltration after early stop- flow procedure without significant destroy occurred in the structure of hepatic lobule. No significant difference of various parameters above occurred between SF and SFC groups.

CONCLUSIONThe hepatic injury following stop- flow procedure was self-limited and reversible. There is no severe destroy of hepatic structure and disfunction during stop- flow chemotherapy.

Animals ; Chemotherapy, Cancer, Regional Perfusion ; methods ; Disease Models, Animal ; Female ; Infusions, Intra-Arterial ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-8 ; metabolism ; Liver ; blood supply ; drug effects ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Swine ; Transcription Factor RelA ; metabolism

OBJECTIVETo investigate amplification of zinc finger protein 217(ZNF217) and its association with clinicopathologic parameters in primary gastric carcinoma.

METHODSSemiquantitative polymerase chain reaction (PCR) was used to determine DNA copies of ZNF217 in the specimens from forty- seven cases with primary gastric carcinoma.

RESULTSThere was no difference in DNA copies between tumor specimens and paratumor normal tissues. The incidence of ZNF217 amplification was 11.36% in gastric cancer. The amplification of ZNF217 was significantly associated with tumor size(P< 0.01) and intestinal type of stomach cancer(P< 0.05).

CONCLUSIONOncogene ZNF217 may play a role in specific tumor types or subtypes of gastric cancer. There may be other oncogenes associated with gastric carcinoma in 20(q)13.

Adult ; Aged ; Aged, 80 and over ; DNA, Neoplasm ; genetics ; Female ; Gene Amplification ; Humans ; Male ; Middle Aged ; Stomach Neoplasms ; genetics ; Trans-Activators ; genetics

OBJECTIVETo determine the microsatellite instability in gastric carcinomas, examine the frameshift mutations of transforming growth factor-beta type II receptor (TGFbetaRII), insulin growth factor II receptor (IGFIIR), bcl-2 associated X protein (BAX) and E2F4, and investigate whether or how alterations of the TGFbetaRII, IGFIIR, BAX and E2F4 gene are associated with MSI in gastric cancer.

METHODSFormalin-fixed, paraffin-embedded gastrectomy specimens and matching normal tissues of 65 cases of gastric carcinomas were retrieved from shanghai Ruijin Hospital and Shanghai East Hospital. DNA was extracted from tissue sections using phenol chloral isoamyl alcohol. Sections with no more than 50% of tumor cell areas were isolated by microdissection. DNA was amplified by PCR-based single strand conformation polymorphism (SSCP) for microsatellite analysis and was sequenced directly. Frameshift mutations in the coding regions, repetitive mononucleotide tracts of (A)10 for TGFbetaRII, (G)8 for IGFIIR, (G)8 for BAX, and trinucleotide repeats of (AGC)13 for transcription factors E2F4 were detected using PCR. Tumors were classified as being microsatellite stable (MSS) or having a low frequency of MSI (MSI-L, one of markers different in the tumor) or a high frequency of MSI (MSI-H, two or more of markers different).

RESULTSEleven cases (18.0%) showed MSI-L, 12 (19.7%) showed MSI-H and 38 (62.3%) cases showed MSS. The mutation rates of TGFbetaRII, IGFIIR, BAX and E2F4 gene were 19.7%, 4.9%, 6.6% and 16.4% respectively. Among the 12 MSI-H gastric cancers, there were 10 TGFbetaRII mutations, 3 IGFIIR mutations, 4 BAX mutations and 10 E2F4 gene mutations. The alterations in the repeats of the related genes presented polymorphisms. Associations of MSI-H status and mutations of the 4 genes were highly significant (P < 0.01, respectively). No repeat tracts mutations in TGFbetaRII, IGFIIR, BAX and E2F4 gene were found in MSS tumors.

CONCLUSIONSThe repeat coding regions within TGFbetaRII, IGFIIR, BAX and E2F4 gene are the targets of microsatellite instability. Frameshift mutations of the 4 genes play an important role in the development and progression of gastric cancers with microsatellite instability.

Adult ; Aged ; Aged, 80 and over ; China ; E2F4 Transcription Factor ; genetics ; Female ; Frameshift Mutation ; Humans ; Male ; Microsatellite Instability ; Middle Aged ; Polymerase Chain Reaction ; Protein-Serine-Threonine Kinases ; Receptor, IGF Type 2 ; genetics ; Receptors, Transforming Growth Factor beta ; genetics ; Stomach Neoplasms ; genetics ; bcl-2-Associated X Protein ; genetics