Objective:To study the function and mechanism of miR-30a in rat cerebral ischemia-reperfusion (I/R) injury.Methods:The middle cerebral artery occlusion (MCAO) model was established by intravascular suture method, and the expression of miR-30a in brain tissue was detected by real-time quantitative polymerase chain reaction (qRT-PCR). After intracerebroventricular injection of miR-30a lentivirus, the infarct area was detected by 2, 3, 5-triphenyltetrazole chloride (TTC) staining, the neurological deficit was detected by Bederson method, and the concentration of neurotrophin-3 (3-NT) and nitric oxide (NO) in brain tissue was detected by enzyme-linked immunosorbent assay (ELISA). The protein levels of kelch like ECH associated protein 1 (Keap1), NF-E2-related factor 2 (Nrf-2) and hemeoxygenase-1 (HO-1) in brain tissue were detected by Western blot. Double luciferase reporter assay was used to detect the targeting relationship between miR-30a and Keap1.Results:Compared with sham operation group, the expression of miR-30a was down-regulated in a time-dependent manner after I/R. The overexpression of miR-30a can reduce the area of cerebral infarction tissue at the pathological level, the degree of neurological impairment at the functional level, the 3-NT, NO and Keap1 at the molecular level, and enhance the expression of Nrf2 and HO-1. The dual luciferase reporter assay also showed that miR-30a could bind to Keap1 mRNA.Conclusions:The expression of miR-30a was down-regulated in MCAO rat brain tissue, and miR-30a could attenuate cerebral I/R injury in rats by targeting Keap1.

Objective To observe the effect of dexamethasone(Dex)on the proliferation and os- teogenic differentiation of human marrow stromal cells(MSCs)in vitro.Methods The primary human MSCs were isolated and cultured by Ficoll seperation culture in vitro.In subcultures,human MSCs were respectively treated with dexamethasone 10~(-9),10~(-8) and 10~(-7) mol/L.The proliferation of human MSCs was measured using MTF method;cytoplasmic alkaline phosphatase(ALP)activity was measured;the osteogenic marker osteopontin (OPN)mRNA were examined by reverse transcriptase polymerase chain reaction(RT-PCR).Results The op- tical density values in cultures treated with dexamethasone 10~(-8) and 10~(-7) mol/L for 8 days were significantly lower than those in the controls(P＜0.05).Treatment of cells with Dex for 12 days led to a significant increase in cytoplasmic ALP activity(P＜0.05)in a dose-dependent manner.Dex induced OPN mRNA.Conclusion Dex inhibits the proliferation of human MSCs and dexamethasone 10~(-7) mol/L leads to a strong decrease in cell number.Dex induces human MSCs differentiate to osteoblastic cells.

OBJECTIVETo analyze the changes of inhibitory killer cell immunoglobulin-like receptors (KIRs), NKG2D receptor and the cytotoxicity of natural killer (NK) cells induced by persistent exposure to CNE2 cells.

METHODSThe HLA-class I genotypes of CNE2 cells and KIR genotypes were determined by PCR with sequence-specific primers (PCR-SSP). The expressions of KIR2DL1, KIR2DL3, KIR3DL1, and NKG2D by the NK cells (freshly isolated NK cells, NK cells cocultured with 100 U/ml IL2 or with 100 U/ml IL2 and CNE2 cells as the control, IL2 and CNE2 groups, respectively) were analyzed by flow cytometry. Cytotoxicity of NK cells against CNE2 cells were detected by LDH releasing assay.

RESULTSThe HLA genotypes of CNE2 cells were A2, 24, B18, 35, Cw4, 7. NK cells isolated from 3 healthy donors expressed KIR2DL1, KIR2DL3, and KIR3DL1. After 4, 24 and 48 h of culture, NK cells in CNE2 group displayed higher KIR2DL1, KIR2DL3 but lower NKG2D expression than those in the control and IL2 groups (P<0.01), whereas the latter two groups showed no significant difference in KIR2DL1, KIR2DL3, and NKG2D expressions (P>0.05), and no difference in KIR3DL1 expression was found between the 3 groups (P>0.05). After 24 h of culture, the cytotoxicity against CNE2 cells mediated by the NK cells in IL2 and CNE2 groups were (26.96-/+1.47) % and (2.74-/+1.64) % at E:T ratios of 10:1, and (35.74-/+3.59)% and (4.57-/+2.41) % at E:T ratio of 20:1, respectively. NK cells in CNE2 group displayed lower cytotoxicity than those in IL2 group (P<0.01).

CONCLUSIONSPersistent exposure to tumor cells expressing NKG2D ligands can lead to downregulated expression of NKG2D receptor, increased expression of KIRs and reduction of NK-mediated cytolysis. These results elucidate the molecular mechanism of reduced cytotoxicity mediated by the edited NK cells and indicate that blocking HLA-class I-bound KIRs or enhancing the expression of NKG2D may promote NK cell-mediated cytolysis.

Cell Line, Tumor ; Cell Survival ; immunology ; Cytotoxicity, Immunologic ; immunology ; Flow Cytometry ; Humans ; Killer Cells, Natural ; cytology ; immunology ; metabolism ; NK Cell Lectin-Like Receptor Subfamily K ; Nasopharyngeal Neoplasms ; immunology ; metabolism ; pathology ; Receptors, Immunologic ; metabolism ; Receptors, KIR ; metabolism ; Receptors, KIR2DL1 ; metabolism ; Receptors, KIR2DL3 ; metabolism ; Receptors, Natural Killer Cell

BACKGROUNDCancer of the esophagus and gastroesophageal junction remains a virulent malignancy with poor prognosis. Rapid progresses were made in chemotherapeutic agents and the development of molecular markers allowed better identification of candidates for targeted therapy. This study aimed to identify the candidate peptides used for anti-angiogenic therapy of esophageal cancer by in vivo screening C7C peptide library for peptides binding specifically to blood vessels of human esophageal cancer.

METHODSThe phage displayed C7C peptide library was injected intravenously into mice bearing human esophageal tumor xenografts under renal capsule. After 5 rounds of screening, 13 clones were picked up individually and sequenced. During each round of screening, titers of phage recovery were calculated from tumor xenograft and control tissues. Homing of these 9 peptides to tumor vessel was detected by calculating phage titers in the tumor xenograft and control tissues (lung and spleen) after each phage was injected into mice model, and compared with the distribution of phage M13 and VIII-related antigen in tumor xenograft by immunohistochemical staining. Comparisons among groups of data were made using one-way analysis of variance (ANOVA), followed by the Bonferroni multiple comparisons test.

RESULTSThe number of phage recovered from tumor tissue of each round increased gradually in tumor group while decreased in control groups (P < 0.01 in tumor and spleen, P < 0.05 in lung). Immunohistochemical staining showed similar staining pattern with M13 antibody or VIII-related antigen antibody, suggesting that phages displaying the selected peptides could home to blood vessel of human esophageal cancer. According to their DNA, 9 corresponding peptide sequences were deduced. And the homing ability to blood vessel of phages displaying the selected peptides was confirmed by comparing with their recovery in tumor and control tissues. Two motifs, YSXNXW and PXNXXN, were also obtained by analyzing the homology of these peptide sequences. The staining distribution of phage with the sequence of PNPNNST was similar to that of the blood vessel marker factor VIII-related antigen staining. After sequencing, each phage with the selected peptide of PNPNNST with 1.0 × 10(11) pfu/ml was injected intravenously into mice. The homing ability to tumor vessel of these 9 kinds of peptides in the xenograft was higher than control tissues (lung and spleen).

CONCLUSIONNine peptides obtained from in vivo screening homed to the blood vessel of human esophageal cancer, and the two motifs of YSXNXW and PXNXXN are the possible biochemical recognition units binding to vascular endothelial cells of esophageal cancer.

Animals ; Antineoplastic Agents ; therapeutic use ; Endothelial Cells ; drug effects ; Esophageal Neoplasms ; blood supply ; drug therapy ; metabolism ; Humans ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Peptide Library ; Peptides ; therapeutic use

OBJECTIVETo explore the association of T1270533G polymorphism in the glutathione S-transferase M1 (GSTM1) gene with the susceptibility to nasopharyngeal carcinoma (NPC) and clinical phenotype of NPC in Chinese population. METHDOS: The genomic DNAs were obtained from 27 Chinese subjects, and the single nucleotide polymorphism (SNP) in all the exons and relevant intron-exon boundaries of GSTM1 were determined by PCR and direct sequencing. A case-control study was performed to analyze the SNP site T1270533G (the rare allele frequency is 22.2% in Chinese population) in the coding region by means of tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) and sequencing.

RESULTSequence analysis identified 29 SNPs in GSTM1 gene, among which 13 SNPs presented high linkage disequilibrium with each other. No obvious relations were found between the variation in the coding region T1270533G and the clinical phenotype of NPC (RR=0.170, 95% CI =0.95-0.306 for TT homozygotes).

CONCLUSIONThe missense mutation in the coding region T1270533G of GSTM1 gene that causes an amino acid change does not affect the detoxification function of GSTM1, and the T1270533G polymorphism does not have apparent relations to NPC susceptibility in Chinese subjects in Guangdong Province.

Adult ; Aged ; Base Sequence ; Carcinoma, Squamous Cell ; genetics ; China ; Female ; Genetic Predisposition to Disease ; genetics ; Glutathione Transferase ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Nasopharyngeal Neoplasms ; genetics ; Polymorphism, Single Nucleotide ; Young Adult

BACKGROUNDMeasurement of human epidermal growth factor receptor 2 (HER2) protein in the serum of metastatic breast cancer patients has previously been reported, but there are no consistent data to support the clinical utility of serum HER2 extracellular domain for patients with early stage breast cancer. We aimed to evaluate the correlation between serum extracellular domain levels and tissue HER2 expression, and analyzed their relationship with clinico-pathological parameters in patients with early stage disease.

METHODSA prospective study was conducted on 232 breast cancer patients with stage I-III prior to treatment. Preoperative serum samples were measured by enzyme-linked immunosorbent assay. Tissue HER2 status was analyzed by immunohistochemistry and fluorescence in situ hybridization assays.

RESULTSThe median serum extracellular domain concentration was 6.8 ng/ml. The best diagnostic cut-off value was 7.4 ng/ml, with 62.9% sensitivity and 85.3% specificity. High serum extracellular domain levels were reported in 89 patients (38.3%), and HER2-positive expression was observed in 77 patients (33.2%). Multivariate analysis showed that elevated serum extracellular domain correlated with postmenopausal status (P < 0.001), high histological grade (P < 0.001), negativity of both estrogen (P = 0.012) and progesterone receptors (P < 0.001), and high levels of carcinoembryonic antigen 153 (P = 0.048).

CONCLUSIONSWe recommend that 7.4 ng/ml should be used as the cut-off value when evaluating serum extracellular domain levels in early stage of breast cancer. Patients with high serum extracellular domain levels have a certain clinico-pathological characteristics, may provide a basis for clinical practice.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; blood ; diagnosis ; Female ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Middle Aged ; Prospective Studies ; Receptor, ErbB-2 ; blood ; metabolism

BACKGROUNDTanis was reported as a putative receptor for serum amyloid A (SAA) involving glucose regulated protein in insulin regulated resistance. It was found to be dysregulated in diabetic rats (Psammomys obesus, Israeli sand rat) and its homologue for humans is SelS/AD-015. The present study analyzed mRNA expression of SelS in omental adipose tissue biopsies from patients with type 2 diabetes mellitus (T2DM), and age- and weight-matched nondiabetic patients, the relationship of SelS mRNA with Homa-IR and serum SAA level.

METHODSHuman omental adipose tissues from ten cases of type 2 diabetic patients and twelve cases of nondiabetic individuals were analyzed for the expression level of SelS mRNA by semiquantitative polymerase chain reaction (PCR), Homa-IR estimated by standard formula and SAA level by enzyme-linked immunosorbent assay (ELISA).

RESULTSSelS mRNA expression, Homa-IR and serum SAA were higher in T2DM sufferers than in nondiabetic control group. SelS mRNA level was positively correlated with Homa-IR and SAA level in each group.

CONCLUSIONSSelS protein may be involved in insulin resistance in Chinese with T2DM by acting as the SAA receptor, thus playing an important role in the development of T2DM and atherosclerosis.

Adipose Tissue ; metabolism ; Adult ; Aged ; Base Sequence ; Diabetes Mellitus, Type 2 ; metabolism ; Female ; Humans ; Insulin Resistance ; Male ; Membrane Proteins ; genetics ; Molecular Sequence Data ; Omentum ; metabolism ; RNA, Messenger ; analysis ; Selenoproteins ; genetics ; Serum Amyloid A Protein ; analysis

OBJECTIVETo investigate the prevalence, etiology and clinical characteristics of adrenal lesions detected by abdominal computed tomography (CT).

METHODSThis retrospective study was conducted in patients with adrenal lesions detected by abdominal CT examinations in Nanfang Hospital between July, 2014 and June, 2015. The clinical data of the patients were collected for analysis of the demographics, comorbidities, imaging characteristics, biochemical profiles, clinical diagnosis and intervention.

RESULTSA total of 939 patients with adrenal lesions were identified from 19 004 patients undergoing abdominal CT scan over the defined period. The mean age of the patients was 53.2 years and 560 of the patients were male. Among the total cases with adrenal lesions, the percentages of cases with adrenal masses tended to increase progressively with age. Endocrine studies were done in 270 of the total patients, which identified non-functioning masses in 38.9%, primary aldosteronism in 16.3%, Cushing's syndrome in 4.1%, subclinical Cushing's syndrome in 7.0%, and pheochromocytomas in 7.0% of the cases. Adrenal incidentalomas was detected in 191 patients, with a detection rate of 1.0% among the overall patients undergoing abdominal CT scans. Imaging study detected adenomas (70.3%), cortical carcinomas (2.4%), and metastases (0.5%). Of 191 patients with adrenal incidentalomas, only 76 (39.8%) underwent endocrine evaluation, including 34 with nonfunctioning adrenal masses, 17 with pheochromocytoma, 7 with primary aldosteronism, and 5 with subclinical Cushing's syndrome.

CONCLUSIONs The overall detection rates of adrenal lesions and adrenal incidentalomas by abdominal CT were 4.9% and 1.0%, respectively, in our cohort of patients undergoing the examination over the defined period. Although most of the lesions were benign and nonfunctioning, malignant and functional lesions were also detected. As many as 60% of the patients with adrenal incidentalomas did not have hormonal testing. Clinicians need to have greater awareness of adrenal incidentalomas and standard protocol for its management should be established.

OBJECTIVETo investigate the value of fluorine-18 fluorodeoxyglucose (FDG)-positron emission tomography (PET) in carcinoma of cardia or fundus of stomach.

METHODSFrom April 1999 to April 2005, 57 patients with carcinoma of cardia or fundus of stomach were imaged with FDG-PET. FDG-PET imaging were analyzed by visual method combined with semiquantitative analysis. The results were compared with pathological findings and follow-up results.

RESULTSIn 29 untreated patients, 25 T(2) to T(4) tumors were all FDG avid and 4 T(1) cases showed nothing abnormal at the primary site. In 24 patients performed curative operation 40 resected enlarged lymph nodes beyond 1 cm were diagnosed correctly by FDG-PET. FDG-PET revealed distant metastases in 5 patients and corrected them from curative surgery candidates to late stage. In 28 treated patients FDG-PET confirmed 22 cases with recurrence or metastasis.

CONCLUSIONSFDG-PET has limited value in confirming T stage in carcinoma of cardia or fundus of stomach. It showed potential in N and M staging and predicting treatment response.

Adult ; Aged ; Aged, 80 and over ; Cardia ; Female ; Fluorodeoxyglucose F18 ; Gastric Fundus ; Humans ; Male ; Middle Aged ; Positron-Emission Tomography ; Radiopharmaceuticals ; Retrospective Studies ; Sensitivity and Specificity ; Stomach Neoplasms ; diagnostic imaging ; pathology