Adult ; Humans ; Male ; Maxillary Sinus ; Xanthogranuloma, Juvenile

?ig-h_3 was first identified as a transforming growth factor-beta1-inducible gene in human lung adenocarcinoma cell line. It encodes for a secreted extracellular matrix (ECM) protein, which is thought to act on cell attachment and ECM composition. Previous study showed that ?ig-h_3 were highly expressed in human hepatoma cell lines and lowly expressed in human normal hepatic cells. The present study aimed to transfect ?ig-h_3 into 7721 cells to investigate its effect on secretion of MMPs in the transfected human hepatoma cells. Full-length ?ig-h_3 gene,cloned by reverse transcription polymerase chain reaction (RT-PCR) was inserted into the eukaryotic expression vector pEGFP-C_2. The recombinant plasmid was transfected into 7721 cells with Lipofectamine2000 and Gelatin-Zymography were adopted to detect the production of MMPs in the transfected cells. Results showed that ?ig-h_3/pEGFP-C_2 recombinant expression plasmid was successfully constructed and achieved high transfection efficiency. MMPs expression of the transfected cells was promoted significantly. These results suggest that overexpression of ?ig-h_3 promoted the production of MMPs, indicating that ?ig-h_3 may play roles in the invasive and metastatic processes of hepatoma.

OBJECTIVETo explore the role of CXCR4/SDF-1alpha axis in organ-specific metastasis of nasopharyngeal carcinoma (NPC) by assessment of CXCR4 expression in NPC cells and SDF-1alpha expression in distant target organs of NPC.

METHODSThirty patients with NPC and fifteen normal subjects were recruited in this study. The expressions of CXCR4 in NPC and normal cases were identified by RT-PCR and immunohistochemistry (IHC), then the relationship between CXCR4 expression and clinicopathological factors was analyzed. IHC was also used to analyze the SDF-1alpha protein expression in normal cervical lymph nodes (including normal superior and inferior deep cervical lymph nodes), bone marrow, lung, liver, kidney and colon tissues of NPC patients (5 cases/each group).

RESULTSThe relative expression level of CXCR4 mRNA in NPC (0.71 +/- 0.22) was significantly higher than that of normal nasopharynx tissues (0.14 +/- 0.07, F = 27.94, P < 0.05). The relative expression level of CXCR4 protein in NPC (1.58 +/- 0.59) was significantly higher than that of normal nasopharynx tissues (0.51 +/- 0.22, F = 17.75, P < 0.05). The high expression levels of CXCR4 mRNA and protein in NPC were closely related to clinical stage, cervical lymph node metastasis and cancer cell differentiation (P < 0.05). SDF-1alpha protein was strongly expressed in normal superior deep cervical lymph nodes, bone marrow, lung and liver (2.35 +/- 0.67), but absent or very poor expression in inferior deep cervical lymph nodes, kidney and colon tissues (0.68 +/- 0.23), and the differences between them were statistically significant (t = 10.13, P < 0.01).

CONCLUSIONCXCR4 is closely correlated to metastasis of nasopharyngeal carcinoma. CXCR4/SDF-1alpha axis may play an important role in organ-specific metastasis of NPC.

Adult ; Aged ; Bone Marrow ; metabolism ; Cell Differentiation ; Chemokine CXCL12 ; genetics ; metabolism ; Female ; Humans ; Liver ; metabolism ; Lung ; metabolism ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Receptors, CXCR4 ; genetics ; metabolism ; Young Adult

Acquired Immunodeficiency Syndrome ; complications ; pathology ; Antigens, CD34 ; metabolism ; Gastrectomy ; methods ; Humans ; Male ; Middle Aged ; Sarcoma, Kaposi ; metabolism ; pathology ; surgery ; virology ; Stomach ; pathology ; Stomach Neoplasms ; metabolism ; pathology ; surgery ; virology ; Vimentin ; metabolism

OBJECTIVEIn order to fully understand hepatitis c virus (HCV) genotype 3b, 1a, 2b and 6a infection in China, We built HCV 5'-noncoding region (5'-NCR) of different genotypes and subtypes.

METHODSThe classification HCV into variable genotypes (subtypes) was carried on by programs A, B and C A. Using a combination of three restriction endonuclease BHH' (BsrB I, Hae II, Hinf I) digestions at the same time. The distinct genotypes were classified into 5 groups: genotype 1 (1a, 1b), 6a, 2 (2a, 2b), genotype 3 (3a, 3b), genotype4 (4a). B. With regard to genotype 1, we could distinguish subtype 1a from 1b using BstU I digestion. C. Using restriction endonuclease Hae III, genotype 2a, 2b, 3b, 4a, 6a are differentiated respectively.

RESULTS(1) HCV genotype 1a, 1b, 2a, 2b, 3a, 3b, 4a, 6a are fully discriminated by comparison with the genotypes regular samples. (2) Of the 93 patients, HCV genotype distribution in China was 66.67% for 1b, 18.28% for 2a, 3.23% for 1b/2b, 3b, 2b respectively. 2.15% for 2a/2b, 1b/2a respectively. 1.08% for 1a.

CONCLUSIONThis research indicated that adoption of HCV 5'-NCR A B C restriction endonuclease digestions techniques, might be sensitive and efficient to detect HCV and discriminate HCV genotype (subtypes) 1a to 6a.

5' Untranslated Regions ; chemistry ; DNA Restriction Enzymes ; Genotype ; Hepacivirus ; classification ; genetics ; RNA, Viral ; analysis

OBJECTIVETo evaluate the effect and acute toxicity of concurrent radio-chemotherapy, by NVB and DDP, plus concurrent radiotherapy in comparison with chemotherapy alone in the treatment of inoperable locally advanced non-small-cell lung cancer (LANSCLC).

METHODSSixty-four patients with inoperable LANSCLC were randomly divided by envelope method into two groups: concurrent radio-chemotherapy group (n = 33) and conventional chemotherapy group (n = 31). The patients in conventional chemotherapy group were treated by NP regimen (NVB + DDP): NVB 25 mg/m(2), d1, 8 and DDP 25-30 mg/m(2) d1-3. In the radio-chemotherapy group by NP regimen plus conventional radiotherapy by (60)Co: 64-68 Gy/2 Gy x 5/w x 6-7w.

RESULTSAll patients completed the treatment schedule. The overall response rate (CR + PR) in the radio-chemotherapy group was significantly higher than that in the conventional chemotherapy group: 81.8% vs 45.2%, (P < 0.01) with 1- and 2-year survival rates of 69.7% vs 38.7% (P < 0.05) and 39.4% vs 16.1% (P < 0.05). without any significant difference in the acute toxicity between two groups (P > 0.05).

CONCLUSIONCompared with conventional chemotherapy (NVP and DDP), concurrent radio-chemotherapy (NVP and DDP plus concurrent radiotherapy) is more effective and tolerable for the inoperable locally advanced non-small-cell lung cancer.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; radiotherapy ; Cisplatin ; administration & dosage ; Combined Modality Therapy ; Female ; Humans ; Lung Neoplasms ; drug therapy ; radiotherapy ; Male ; Middle Aged ; Radiation-Sensitizing Agents ; administration & dosage ; Vinblastine ; administration & dosage ; analogs & derivatives

Objective To establish a high throughput targeted method to study phospholipids profding and to screen out and quantitate the potential biomarkers in urine samples.Methods The phospholipids in urine was extracted by modified MTBE(methyl tert-butyl ether)method.Semi -quantitative analysis of phospholipids in urine was realized by using high performance liquid chromatography-electrospray ionization-qua-drupoles/trap (HPLC-ESI-Q/Trap) technique.7 standards and 15 endogenous phospholipids were chosen to conduct method validation,including specificity,sensitivity,precision,matrix effect,carryover effect,stability and recovery.Principal component analysis (PCA) of quality control (QC) samples interspersed during the detection process was used to evaluate the reliability of the data obtained.Results The lower limit of quantitation of 7 phospholipids were phosphatidylcholine and lysophosphatidylcholine 0.25 ng· mL-1,phosphatidylethanolamine and phosphatidylserine 2.00 ng· mL-1,phosphatidylglycerol and phosphatidylinositol 1.00 ng · mL-1,sphingomyelin 625.00 pg · mL-1,respectively.During the continuous analysis,the relative standard deviation (RSD) of the retention time was 0.72％-3.44％,the peak area was 0.71％-10.53％.The recoveries of the 7 phospholipids were in the range of 54.05％-105.73％.All samples were stable after being stored 12 h at room temperature,being stored 24h after preparation,two freeze-thaw cycles and being cryopreserved 1 month at-80 ℃.QC samples in the first principal component diagram showed that the data was reliable.Conclusion The developed HPLC-ESI-Q/Trap method was simple,stable and sensitive,which can be applied to the subsequent study of large sample size of phospholipidomics research and quantitative analysis of potential urinary phospholipids biomarkers.

Objective To investigate the effects of the enhancer of zeste homolog 2 (EZH2) gene on cell growth and invasion of the nasopharyngeal carcinoma (NPC).Methods Recombinant lentivirus vector for shRNA delivery of EZH2 was constructed and transfected into 293FT cells. After collecting the viral particles, the NPC cell line 5-8F cells were transfected. The effects of EZH2 silence on cell proliferation and cell cycle were detected using MTT assay,plate colony formation assay and flow cytometry.The migration and invasion of 5-8F cells were determined by wound healing assay and matrigel invasion assay,respectively.The expressions of EZH2 and epithelial-mesenchymal transition (EMT) -related markers at mRNA and protein levels were examined by real-time PCR and Western blot respectively.Results The expressions of EZH2 mRNA and protein in the transfected 5-8F cells were obviously reduced. MTT assay showed that EZH2 downregulation significantly inhibited the growth of 5-8F/shEZH2 cells( P ＜ 0.001 ).Colony formation rate (84.44％ ) of 5-8F/shEZH2 cells was lower than control (31.56％,P =0.001 ).Cell cycle analysis showed that most 5-8F/shEZH2 cells were arrested in G0/G1 phase,with a very low ratio of cells in S phase.Wound healing assay indicated that the migration ability of cells silencing EZH2 decreased significantly,and the 48-hour relative migration distance of 5-SF/ShEZH2 cells and control cells was 0.58 ± 0.05,and 0.81 ± 0.02,respecptively(P ＜ 0.000).Matrigel invasion assay,showed the invasive capacity of cells silencing EZH2 was significantly inhibited,with less penetrating cells (72.23 ± 4.08 ) compared to control( 150.95 ± 16.27),P ＜ 0.000.The mRNA expressions of epithelial markers E-cadherin and Keratin 18 in the cells silencing EZH2 increased by 177％ and 158％ respectively,and the mRNA expressions of mesenchymal markers β-catenin and N-cadherin decreased by 18.04％ and 41.18％ respectively. Similar results also were obtained with Western blot analysis. Conclusion EZH2 significantly enhanced the proliferation and invasion of nasopharyngeal carcinoma cells in vitro,which might be mediated by inducing EMT.

OBJECTIVETo observe the effect of Shouwu-Huanjing Recipe(SWHJR) medicated serum on human sperm motility and fertility in vitro.

METHODSHuman sperm was co-cultured with SWHJR medicated serum in vitro. Human sperm motility was evaluated by computer-assisted semen analysis(CASA). The acrosome reaction and the capability of penetrating zona-free hamster eggs were also observed.

RESULTSThe co-cultured SWHJR medicated serum significantly increased the sperm motion velocity(VAP, VCL, VSL) (P < 0.01), the amplitude of lateral head movement (ALH) and the beat frequency of flagellum(BCF), the density of progressive motility sperms (P < 0.05), the acrosome reaction rate(P < 0.001), the fertilization rate(FR) and the fertilization index (FI) in sperm penetration assay(SPA) test (P < 0.01). The stimulation of SWHJR medicated serum occurred in dose-dependent manner.

CONCLUSIONSWHJR can improve human sperm motility and fertility.

Acrosome Reaction ; Animals ; Cricetinae ; Fertility ; drug effects ; Humans ; Male ; Medicine, Chinese Traditional ; Mesocricetus ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; drug effects

OBJECTIVETo investigate the relationship of angiogenesis and the Ang family members/ receptor (Ang/Tie2) in hemangioma.

METHODSExpression of Ang1, Ang2 and the receptor Tie2 was detected with immunohistochemical SP method and RT-PCR method in 17 cases of proliferating hemangioma, 13 involuting cases and 10 cases of normal children skin.

RESULTSThe expression of Ang2 and Tie2 was higher markedly in proliferating hemangiomas than in involuting hemangiomas (P < 0.01), and was rare or negative in normal skin. Expression of Ang1 was rare or negative both in hemangioma and normal skin without significant difference between them (P > 0.05).

CONCLUSIONAng/Tie2 system may play an important role in the proliferating and involuting process of hemangioma.

Angiopoietin-1 ; metabolism ; Angiopoietin-2 ; metabolism ; Child, Preschool ; Hemangioma ; metabolism ; pathology ; Humans ; Infant ; Receptor, TIE-2 ; metabolism