Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Infant ; Male ; Thrombasthenia ; classification ; diagnosis

Studies have shown cell-free microRNA (miRNA) circulating in the serum and plasma with specific expression in cancer, indicating the potential of using miRNAs as biomarkers for cancer diagnosis and therapy. This study was to investigate whether plasma miRNA-21 (miR-21) can be used as a biomarker for the early detection of non-small cell lung cancer (NSCLC) and to explore its association with clinicopathologic features and sensitivity to platinum-based chemotherapy. We used real-time RT-PCR to investigate the expression of miR-21 in the plasma of 63 NSCLC patients and 30 healthy controls and correlated the findings with early diagnosis, pathologic parameters, and treatment. Thirty-five patients (stages IIIB and IV) were evaluable for chemotherapeutic responses: 11 had partial response (PR); 24 had stable and progressive disease (SD+ PD). Plasma miR-21 was significantly higher in NSCLC patients than in age- and sex-matched controls (P < 0.001). miR-21 was related to TNM stage (P < 0.001), but not related to age, sex, smoking status, histological classification, lymph node status, and metastasis (all P > 0.05). This marker yielded a receiver operating characteristic (ROC) curve area of 0.775 (95% CI: 0.681- 0.868) with 76.2% sensitivity and 70.0% specificity. Importantly, miR-21 plasma levels in PR samples were several folds lower than that in SD plus PD samples (P = 0.049), and were close to that in healthy controls (P = 0.130). Plasma miR-21 can serve as a circulating tumor biomarker for the early diagnosis of NSCLC and is related to the sensitivity to platinum-base chemotherapy.
Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; blood ; Carboplatin ; administration & dosage ; Carcinoma, Non-Small-Cell Lung ; blood ; drug therapy ; pathology ; Cisplatin ; administration & dosage ; Early Detection of Cancer ; Female ; Humans ; Lung Neoplasms ; blood ; drug therapy ; pathology ; Male ; MicroRNAs ; blood ; Middle Aged ; Neoplasm Staging ; Remission Induction

OBJECTIVETo investigate the expression of forkhead box M1 (FOXM1) and its correlation with clinicopathological features and prognosis in patients with non-small cell lung cancer (NSCLC).

METHODSThe expression of FOXM1 in 68 cases of NSCLC was detected by immunohistochemistry. The FOXM1 expression in 6 tumor tissues (3 cases with negative and 3 cases with positive expression of FOXM1) was analyzed by Western blotting to confirm the immunohistochemical results. The correlation of the expression of FOXM1 with clinicopathalogical features and overall survival of the NSCLC patients was analyzed.

RESULTSThe expression of FOXM1 protein was detected in the nuclei or cytoplasms of the tumor cells. The positive expression rate of FOXM1 was 36.8% (25/68). Western blotting confirmed the immunohistochemical results. The expression level of FOXM1 in advanced stage cancer was significantly higher than that in early stage NSCLC (P = 0.001). The median OS was 23.0 months in patients with negative expression of FOXM1 and 13.0 months in those with positive expression (P = 0.001). Univariate analysis revealed that histological grade, lymph nodes status, TNM stage and FOXM1 expression were significantly associated with prognosis in the NSCLC patients (P < 0.05). The Cox multivariate analysis demonstrated that lymph nodes status, TNM stage and FOXM1 expression were independent poor prognostic factors (P < 0.05).

CONCLUSIONThe expression status of FOXM1 in NSCLC is an independent prognostic factor and negatively correlated with prognosis.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Follow-Up Studies ; Forkhead Box Protein M1 ; Forkhead Transcription Factors ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Grading ; Neoplasm Staging ; Proportional Hazards Models

OBJECTIVETo assess the expression of HER-2 and leptin in gastric cancer and evaluate their relationship with VEGF expression and clinicopathological features, and their prognostic value for gastric cancer patients.

METHODSOne hundred and ten gastric cancer specimens and the corresponding metastatic lymph nodes were detected for HER-2 by immunohistochemistry (IHC). All primary cancer tissues were detected for leptin, OB-Rb and VEGF. Ninty-six specimens of normal gastric mucosa served as the control.

RESULTSThe expression level of HER-2, leptin and OB-Rb in gastric cancer tissues were significantly higher than those in normal tissues (19.1% vs. 8.0%, 49.1% vs. 34.0%, and 60.9% vs. 46.0%, P < 0.05). HER-2 overexpression was moderately homogenous in primary gastric cancer and matastatic lymph nodes (P = 0.607, Kappa = 0.581). There was a correlation between the expression of HER-2 and leptin, both of which were significantly correlated with tumor invasion depth, metastatic lymph nodes ratio (NR), distal metastasis, TNM stage and VEGF expression. However, there was no significant correlation between OB-Rb expression and the clinicopathological features evaluated. Cox regression multivariate analysis showed that tumor size, histological grade, NR, stage, chemotherapy and HER-2 expression were independent prognostic factors.

CONCLUSIONSHER-2 is stably expressed in primary gastric cancer and metastatic lymph nodes. HER-2 and leptin play an important role in the progression and angiogenesis of gastric cancer. High expression of HER-2 is a prognostic factor for poor outcome.

Adenocarcinoma ; drug therapy ; metabolism ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Chemotherapy, Adjuvant ; Disease-Free Survival ; Female ; Follow-Up Studies ; Gastrectomy ; Humans ; Leptin ; metabolism ; Lymph Nodes ; metabolism ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Proportional Hazards Models ; Receptor, ErbB-2 ; metabolism ; Receptors, Leptin ; metabolism ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Survival Rate ; Tumor Burden ; Vascular Endothelial Growth Factor A ; metabolism

BACKGROUNDExcessive deposition of extracellular matrix (ECM) in the kidney is the hallmark of diabetic nephropathy. Increased matrix synthesis has been well documented but the effects of diabetes on degradative pathways, particularly in the in vivo setting. The renal protective effect of these pathways on matrix accumulation has not been fully elucidated. The present study was undertaken to investigate the activity of matrix metalloproteinase-2 (MMP-2), the expression of MMP-2 and tissue inhibitor of metalloproteinase-2 (TIMP-2) in kidney tissues of diabetic rats, and to explore the degradative pathway of type IV collagen (IV-C) and the renal protective effects of ACE inhibition-benazepril.

METHODSTwenty-four healthy male Wistar rats were divided randomly into normal control group (NC group), untreated diabetes mellitus group (DM group), and diabetes mellitus group treated with benazepril (DL group). The rat model of diabetes mellitus was induced by intraperitoneal injection of streptozocin (60 mg/kg). After the establishment of DM model, benazepril (10 mg.kg(-1).d(-1)) was given to the DL group for 12 weeks, and the same volume of water was given to the other two groups. At the end of 12 weeks, renal function was evaluated with 24-hour urinary protein (Upro), clearance of creatinine (Ccr), and blood urea nitrogen (BUN). MMP-2 activity was determined by gelatin zymography. The levels of MMP-2, TIMP-2 and collagen IV (IV-C) protein in the kidney tissue were assessed by immunohistochemistry. The gene expression of MMP-2 and TIMP-2 was measured by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSThe levels of BUN, Upro and Ccr in the DM group were higher than those in the NC group. In the DM group, the mRNA, enzymatic activity and proteins of MMP-2 decreased, but the expressions of IV-C and TIMP-2 increased. All diabetes-associated changes in renal function and MMP/TIMP were attenuated after benazepril treatment with reduced IV-C accumulation.

CONCLUSIONSThe changes of MMP-2 and TIMP-2 expressions in kidney tissues of diabetes rats may contribute to the occurrence and progression of diabetic nephropathy. Benazepril could exert protective effects on diabetic nephropathy, owing to the upregulation of MMP-2 and downregulation of TIMP-2 expressions, which further inhibits the excessive deposition of extracellular matrix in the glomerulus.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Animals ; Benzazepines ; pharmacology ; Blood Glucose ; analysis ; Body Weight ; Collagen Type IV ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Diabetic Nephropathies ; prevention & control ; Kidney ; drug effects ; metabolism ; Kidney Glomerulus ; pathology ; Male ; Matrix Metalloproteinase 2 ; analysis ; genetics ; Organ Size ; Rats ; Rats, Wistar ; Streptozocin ; Tissue Inhibitor of Metalloproteinase-2 ; analysis ; genetics

Objective To analyze the efficacy and biomechanical properties of locking proximal femoral plate (LPFP) and proximal femoral nail anti-rotation (PFNA) for treating intertrochanteric fracture in elderly patients. Methods One hundred and six elderly patients with intertrochanteric fracture of femur were randomly divided into LPFP group (53 cases) and PFNA group (53 cases). After treatment intervention, the operation time, intraoperative bleeding volume, weight-bearing time, fracture healing time and Harris hip function score of 9 months after operation in two groups were recorded. The complications after operation in two groups were analyzed. Ten elderly fresh femoral specimens were selected to prepare the model of intertrochanteric femoral fracture in the elderly. They were randomly divided into PFNA group and LPFP group with 5 models in each group. After treatment and intervention, axial compression test, destructive load test and torsional stiffness test were conducted by mechanical testing machine, and biomechanical properties were recorded. Results The average operation time, weight-bearing time and fracture healing time in PFNA group were shorter than those in LPFP group (P<0.05), and the average intraoperative bleeding volume in PFNA group was less than that in LPFP group (P<0.05), and the average Harris score was higher than that in LPFP group (P<0.05). The total incidence of postoperative complications in PFNA group and LPFP group was 7.56% and 18.87%, respectively, indicating a significant difference between the two groups (P<0.05). After intervention treatment, the average axial compression, damage load and torsional stiffness in PFNA group were higher than those in LPFP group (P<0.05). ConclusionsPFNA caused minimal trauma for treating intertrochanteric femoral fractures in the elderly. With its good biomechanical properties, PFNA could effectively promote fracture healing and hip function recovery, and significantly reduce the incidence of hip varus, screw loosening and cutting complications.

OBJECTIVETo observe effects of removing arms and ovarian on lumbar intervertebral disc and vertebral bone mineral density (BMD) by establishing rat model of lumbar intervetebral disc degeneration (IDD) with kidney deficiency, and to explore internal mechanism of disc degeneration, relationship between disc degeneration and osteoporosis.

METHODSThirty Sprague-Dawley female rats aged one month were randomly divided into control group, lumbar IDD group and lumbar IDD with kidney deficiency group (combined group), 10 rats in each group. Lumbar IDD group removed double arms, lumbar IDD with kidney deficiency group removed double arms after 3 months, both ovaries were removed. Vertebral bone mineral density were observed by Micro-CT scan; morphological changes were tested by safranine O-fast green staining; II, X collagen protein expression in the intervertebral disc were obsevered by immunohistochemistry; extracellular matrix gene expression were obsevered by real-time polymerase chain reaction (RT-PCR), in order to evaluate the effects of removed of forelimbs and double ovarian on degeneration and vertebral bone mineral density of intervertebral disc.

RESULTSMicro-CT scan showed osteoporosis in kidney deficiency group was obviously worse than other two groups; safranine O-fast green staining showed that intervertebral space became narrowed, intervertebral disc tissue degenerated obviously, chondral palte was underdeveloped in kidney deficiency group; immunohistochemistry showed that X collagen expression increased, type II collagen expression decreased in kidney deficiency group; RT-PCR showed that type II collagen expression in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P=0.000); Age 1 in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P= 0.000); while type X collagen expression was higher than control group, but no significant meaning; MMP-13 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P= 0.000, P=0.000); aggrecanase-2 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P=0.006, P=0.008).

CONCLUSIONRats model of lumbar disc degeneration established by removed forelimbs and ovariectomized can occure "bone like"--osteoporosis, which is similar with clinical kidney lumbar disc degeneration in tissue morphology, molecular cell biology expression.

Animals ; Collagen ; genetics ; metabolism ; Extracellular Matrix ; genetics ; metabolism ; Female ; Humans ; Intervertebral Disc Degeneration ; etiology ; metabolism ; physiopathology ; surgery ; Kidney ; physiopathology ; Osteoporosis ; complications ; genetics ; metabolism ; Ovariectomy ; adverse effects ; Rats ; Rats, Sprague-Dawley

BACKGROUNDChina is one of the countries with the highest incidence of H. pylori and more than 9090 isolates possessed the cagA gene. This study was to evaluate the biological activity of the H. pylori virulence factor cagA isolated from Chinese patients.

METHODScagA DNA fragments were amplified from the genomic DNA and subsequently cloned into the mammalian expression vector for cell transfection and DNA sequencing. cagA protein, phosphorylated-tyrosine cagA and the complex of cagA precipitated with SHP-2 were identified respectively by western blot in the crude cell lysate from conditionally immortalized gastric epithelial cells at 48 hours after transfection with cagA DNA. In addition, the ability of induction of scattering phenotype was examined after transient expression of cagA in AGS cells.

RESULTSThe C-terminal half of cagA contained only one repeated sequence and three tandem five-amino-acid motifs glutamic acid-proline-isoleucine-tyrosine-alanine (EPIYA). Moreover, the amino acid sequence of D2 region in repeated sequence was aspartic acid-phenylanaline-aspartic acid (D-F-D) which was significantly distinguished from the three repeated sequences and aspartic acid-aspartic adid-leucine (D-D-L) in the western standard strain NCTC11637. Western blot revealed that cagA became phosphorylated in tyrosine site and bound with SHP-2 after transient expression of cagA DNA in gastric epithelial cells. Transient expression of cagA in AGS cells showed that cagA was able to induce the elongation phenotype although to a lesser extent than western strains.

CONCLUSIONScagA perturbs cell signaling pathways by binding with SHP-2. However, significant difference exists in amino acid sequence and biological function of cagA in Chinese compared with those of western countries.

Amino Acid Sequence ; Antigens, Bacterial ; chemistry ; physiology ; Bacterial Proteins ; chemistry ; physiology ; Blotting, Western ; Cells, Cultured ; Gastric Mucosa ; Humans ; Intracellular Signaling Peptides and Proteins ; Molecular Sequence Data ; Phenotype ; Phosphorylation ; Protein Tyrosine Phosphatase, Non-Receptor Type 11 ; Protein Tyrosine Phosphatases ; metabolism ; Repetitive Sequences, Amino Acid ; Signal Transduction

The purpose of this study was to investigate the influence of immunosuppressive therapy on the expression of TNF-α/IFN-γ in cytoplasm of peripheral blood lymphocytes of patients with aplastic anemia (AA). The expression of TNF-α and IFN-γ in cytoplasm of peripheral CD3(+) lymphocytes were measured by flow cytometry in 25 cases of de novo AA patients and 20 cases of AA after immunosuppressive therapy. The results showed that the positive rates of CD3(+)/TNF-α(+) and CD3(+)/IFN-γ(+) in de novo AA patients were (5.97 ± 6.78)% and (15.20 ± 11.28)% respectively, and (1.56 ± 0.87)% and (1.76 ± 0.87)% in normal controls respectively. There was significant difference between de novo AA patients and normal controls (p < 0.05). The positive rates of CD3(+)/TNF-α(+) and CD3(+)/IFN-γ(+) in immunosuppressive therapy group were (1.67 ± 1.26)% and (4.35 ± 4.33)% respectively. The difference between immunosuppressive therapy group and de novo AA group was statistically significant (p < 0.05). It is concluded that the levels of intracellular TNF-α and IFN-γ in AA patients are higher than those in normal controls. Immunosuppressive therapy significantly reduces the expression of intracellular TNF-α and IFN-γ. Its relationship with the clinical treatment is worth further observing.
Adolescent ; Adult ; Anemia, Aplastic ; blood ; metabolism ; therapy ; Female ; Humans ; Immunosuppression ; Interferon-gamma ; metabolism ; Lymphocytes ; metabolism ; Middle Aged ; Tumor Necrosis Factor-alpha ; metabolism ; Young Adult

OBJECTIVETo investigate the efficacy and safety of chemotherapy combined with intraperitoneal perfusion of cytokine-induced killer (CIK) cells for advanced gastric cancer patients with ascites.

METHODSFrom January 2008 to December 2010, 42 advanced gastric cancer patients with ascites in Zhongshan Hospital of Fudan University were enrolled in the study. According to personal choice, patients were divided into 2 groups: XELOX chemotherapy alone (Capecitabine and Oxaliplatin) was applied in 22 patients (chemotherapy group) and XELOX combined with intraperitoneal perfusion of CIK cells in 20 patients (combination group). The efficacy, safety, and immunological function, including the time to progression (TTP), overall survival (OS), Karnofsky performance status (KPS) score, immunity index (CD4+/CD8+ ratio), volume of peritoneal fluid, were compared between two groups.

RESULTSCompared with the chemotherapy group after treatment, the combination group had a higher KPS score (78.0±9.8 vs. 70.0±8.9, P=0.009), less volume of 2-cycle peritoneal fluid drainage [(4500±1218) ml vs. (5527±1460) ml, P=0.018 ], longer median TTP (4.0 vs. 2.5 months, P=0.001) and OS (11.0 vs. 6.0 months, P=0.006), higher ratio of CD4+/CD8+ (1.34±0.36 vs. 0.96±0.26, P=0.001). While no significant significances were found between the two groups in disease response rate (35.0% vs. 22.7%, P=0.499) and disease control rate (75.0% vs. 54.5%, P=0.209). There were no serious adverse reactions in the combination group.

CONCLUSIONSAs compared with XELOX chemotherapy alone, the combination immunological treatment of XELOX chemotherapy and intraperitoneal perfusion of CIK cells possesses better efficacy for the advanced gastric cancer patients with ascites, which can prolong the survival and enhance the immunological function with favorable safety.

Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; Ascites ; etiology ; therapy ; Combined Modality Therapy ; Cytokine-Induced Killer Cells ; Deoxycytidine ; analogs & derivatives ; Female ; Fluorouracil ; analogs & derivatives ; Humans ; Immunotherapy, Adoptive ; Male ; Middle Aged ; Pilot Projects ; Stomach Neoplasms ; complications ; therapy ; Treatment Outcome