Carcinoma ; genetics ; metabolism ; pathology ; surgery ; China ; Humans ; Male ; Oncogene Proteins, Fusion ; genetics ; Prostatic Hyperplasia ; genetics ; metabolism ; pathology ; surgery ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Proto-Oncogene Proteins c-ets ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serine Endopeptidases ; genetics ; metabolism

BACKGROUNDOur previous research has suggested that platelet activating factor receptor was related to atherosclerosis. The present study investigated the effect of a platelet activating factor receptor antagonist-WEB2086 on angiogenesis in aortal plaque and ischemic hindlimb of apolipoprotein E-deficient mice.

METHODSEight-week-old apolipoprotein E-deficient mice were fed with a 0.15% cholesterol diet to develop advanced lesions. At age 32 weeks unilateral hindlimb ischemia was surgically induced and the mice were divided into two groups: with or without WEB2086 mixed with their drinking water (4.3 mg in 100 ml). At age 40 weeks blood was collected from the orbit for measurement of serum lipids and an enzyme linked immunosorbent assay was used to determine platelet activating factor and oxidized low density lipoprotein in the gastrocnemius and aorta. Whole-Mount CD31 stain and plaque-associated sprouting have been used to estimate angiogenesis in plaque from the aorta and laser Doppler perfusion imaging and immunohistochemical expression of von Willebrand factor have been used to estimate angiogenesis in ischemic hindlimb.

RESULTSThe lipid composition of serum was not different between the groups. However, the amount of platelet activating factor and oxidized low density lipoprotein detected in the aorta was significantly higher than that in the gastrocnemius of ischemic hindlimb. The ratio of lesion to aorta levels was significantly reduced by administration of WEB2086, (31.52 +/- 6.18)% vs (55.58 +/- 8.34)%, P < 0.01. The mean density of intimal capillaries in atherosclerotic plaque, (31.13 +/- 9.20)% vs (57.74 +/- 11.28)%, P < 0.01, and the mean number of sprouts per aorta were significantly reduced, 183.92 +/- 34.17 vs 392.54 +/- 76.79, P < 0.01, in the WEB2086 group. Blood flow (0.85 +/- 0.12 vs 0.45 +/- 0.06, P < 0.01) and capillary density of ischemic hindlimb (1.18 +/- 0.17 vs 0.53 +/- 0.09, P < 0.01) were markedly increased in apolipoprotein E-deficient mice treated with WEB2086 versus controls.

CONCLUSIONThe study provides evidence that WEB2086 can inhibit angiogenesis in atherosclerotic plaque but promote it in ischemic hindlimb.

Animals ; Apolipoproteins E ; deficiency ; Atherosclerosis ; physiopathology ; Azepines ; pharmacology ; Capillaries ; Cholesterol ; blood ; Hindlimb ; blood supply ; Ischemia ; physiopathology ; Lipoproteins, LDL ; blood ; physiology ; Male ; Mice ; Neovascularization, Physiologic ; drug effects ; Platelet Activating Factor ; analysis ; Platelet Membrane Glycoproteins ; antagonists & inhibitors ; Receptors, G-Protein-Coupled ; antagonists & inhibitors ; Triazoles ; pharmacology

OBJECTIVETo detect the serum proteomic fingerprints in patients with hypopharyngeal squamous cell carcinoma (HPSCC) by surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) protein chip array technique.

METHODSThe serum samples were obtained from 58 HPSCC patients for protein expression analysis using SELDI-TOF Protein Chip technique and cation-exchange (CM10) protein array. All the spectra were compared and the qualified mass peaks with mass-to-charge ratios (m/z) between 1 and 70 kD were autotimatically detected. The tree analysis pattern was generated using Biomarker Patterns Software.

RESULTSThe protein profiles of HPSCC serum were analyzed according to the clinical and pathological features of the patients and their treatment response. No significant difference was noted in the serum proteins between HPSCC patients with different statuses of cervical lympha node metastasis (P>0.05), and the difference between well differentiated and poorly differentiated HPSCC was only minor. No significant difference was found in the serum proteins between chemotherapy-sensitive patients and the insensitive patients (P>0.05), but 5 proteins were identified to be overexpressed in the sensitive patients (P < / = 0.05). Radiotherapy-sensitive HPSCC patients were segregated from the insensitive group with a sensitivity of 86.67% and specificity of 100%.

CONCLUSIONThe serum protein at the m/z value of 6115.74 is overexpressed in radiotherapy-sensitive HPSCC patients. Serum protein profiling allows the prediction of radiotherapy response in HPSCC patients, and the identified proteins may serve as candidate biomarkers for predicting the radiotherapy sensitivity of HPSCC.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; analysis ; Carcinoma, Squamous Cell ; genetics ; radiotherapy ; Female ; Humans ; Hypopharyngeal Neoplasms ; genetics ; radiotherapy ; Male ; Middle Aged ; Models, Biological ; Proteome ; analysis ; Radiation Tolerance ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

OBJECTIVETo explore the influence of oxidized high-density lipoprotein (oxHDL) on the maturation and migration of bone marrow-derived dendritic cells (BMDCs) from C57BL/6J mice.

METHODSThe C57BL/6J mice bone marrow cell suspension was prepared and purified. Recombinant granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant interleukin-4 (rmIL-4) were used to promote monocytes to differentiate and suppress lymphocytes. Then 50 microg/mL oxHDL was added to stimulate BMDCs, using 50 microg/mL high-density lipoprotein (HDL) as homologous protein control, PBS as negative control, and 1 microg/mL lipopolysaccharide (LPS) as positive control. The CD86 and MHCII expression rates were detected with fluorescence-activated cell sorting (FACS). Liquid scintillation counting (LSC) was used in mixed lymphocyte reactions (MLRs) to reflect the ability of BMDCs in stimulating the proliferation of homologous T cells. Levels of cytokines IL-12 and IL-10 were detected by ELISA. The cell migration was evaluated with the transwell system.

RESULTSCompared with PBS group, the expressions of CD86 and MHCII, counts per minute of MLRs, secretion of IL-12 and IL-10, and number of migrated cells in oxHDL group and LPS group significantly increased (all P<0.05), while the increment was less in oxHDL group than LPS group. The number of migrated cells in oxHDL group was about twice of that in HDL group.

CONCLUSIONOxHDL may promote the maturation and migration of BMDCs in vitro.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; physiology ; Cell Differentiation ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; drug effects ; physiology ; Humans ; Lipoproteins, HDL ; metabolism ; pharmacology ; Lipoproteins, LDL ; metabolism ; pharmacology ; Mice ; Mice, Inbred C57BL

OBJECTIVETo determine the value of concurrent chemoradiotherapy in patients with locoregionally advanced nasopharyngeal carcinoma from the mainland of China.

METHODSData were extracted from randomized trials comparing chemotherapy plus radiotherapy with radiotherapy alone in locally advanced nasopharyngeal carcinoma. Actuarial rates of survival and distant metastases were calculated. The followed electronic databases were searched the Chinese Biomedicine database, Pubmed, Medline, Embase and Cochrane library; Data were extracted by tow reviewers and Review manager 4.1 software was applied for statistical analysis.

RESULTSEighteen trials with 1993 patients were include according to the including criterion. The 3-year overall survival rate of the chemoradiotherapy group and the radiotherapy group were 68.47% and 56.38% respectively, and the 5-year overall survival rate of the two groups above were 51.91% and 41.09% respectively, while the distant metastases rate of the chemoradiotherapy group and the radiotherapy group were 26.19% and 38.71% respectively. The result demonstrated that chemoradiotherapy increased overall survival by 12% at 3 years, and 11% at 5 years after treatment. After chemoradiotherapy, the rate of distant metastasis was reduce by 12%.

CONCLUSIONSIn patients with locoregionally advanced nasopharyngeal carcinoma, chemoradiotherapy significantly improves overall survival at 3 years, and 5 years compared with radiotherapy alone.

Antineoplastic Combined Chemotherapy Protocols ; China ; Combined Modality Therapy ; Humans ; Nasopharyngeal Neoplasms ; drug therapy ; radiotherapy ; Radiotherapy ; Randomized Controlled Trials as Topic ; Survival Rate

OBJECTIVESTo investigate the effect of four types of neck dissections for the recurrent and persistent lymph nodes of NPC after radiotherapy.

METHODSThe clinical data of 88 cases of nasopharyngeal carcinoma with recurrent and persistent lymph nodes after radiotherapy were analyzed retrospectively, the 5-year survival rate, recurrent rate, distant metastatic rate and surgical complications were analyzed. The survival rate and recurrent rate of the radical neck dissection (RND), modified radical neck dissection (MRND), selective neck dissection (SND) and lymph node resection (LNR) were compared. The survival and recurrent rate between those with and without postoperative radiotherapy were investigated as well.

RESULTSThe 5-year survival rate and recurrent rate of whole group were 42.8% and 22.7%, respectively. As for the patients with disease staged II, III, IV, the 5-year survival rates were 56.7%, 36.1% and 32.4%, respectively. The 5-year survival rate of groups of RND, MRND, SND and LMR were 39.8%, 60.0%, 37.9% and 44.1%, respectively, the differences were insignificant (Log Rank = 1.0, P = 0.8011), the recurrent rate of the lymph node among the 4 groups were insignificant either (chi2 = 0.470, P = 0.493). The 5-year survival rates of those with and without postoperative radiotherapy were 39.1% and 45.3%, respectively, the differences were insignificant (Log Rank = 0.06, P = 0.8138), the recurrent rate of the two groups were insignificant (chi2 = 0.593, P = 0.441).

CONCLUSIONSThe four types of neck dissection were effective and safe to control the recurrent and persistent lymph nodes in the neck after radiotherapy, as long as choosing patients rationally and gave postoperative radiotherapy if necessary.

Adult ; Female ; Humans ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; pathology ; surgery ; Neck Dissection ; methods ; Neoplasm Recurrence, Local ; surgery ; Neoplasm, Residual ; surgery ; Treatment Outcome

BACKGROUNDIn patients suffering from acute pancreatitis, the pathogenesis is not completely understood, and several recent studies in vitro suggested that heat shock proteins might play an important role in cell signaling. To investigate the possible role of extracellular heat shock protein 70 (Hsp70) in pancreatitis, toll-like receptor-4 (TLR4)-deficient and wild-type mice were administered with exogenous Hsp70 during the course of cerulein-induced pancreatitis (CIP).

METHODSAcute pancreatitis was induced by 5 intraperitoneal injections of cerulein at hourly intervals, and then treated with recombinant Hsp70 through the caudal vein 4 hours after the start of cerulein injections. Subsequently serum amylase and serum cytokines levels were detected. Histologic alteration of the pancreas was evaluated. Tumor necrosis factor alpha (TNF-alpha) concentrations and myeloperoxidase (MPO) activity in both pancreas and lungs were analyzed. The nuclear factor kappa B (NF-kappaB) activation in pancreatic tissue was measured using a sensitive RelA enzyme-linked immunosorbent assay.

RESULTSTreatment with recombinant Hsp70 to wild-type mice in CIP resulted in significant aggravation of inflammation in pancreas, elevated levels of serum cytokines, up-regulation of pulmonary MPO activity and increase of lung tissues TNF-alpha concentrations. In contrast, treatment with Hsp70 to TLR4-deficient mice had little effect on serum cytokines levels, pancreatic inflammation, pulmonary MPO activity and TNF-alpha concentrations.

CONCLUSIONSThe results suggest that extracellular Hsp70 might induce systemic inflammatory response syndrome (SIRS)-like response in vivo and TLR4 might be involved in the Hsp70-mediated activation of inflammatory reaction in the progression of CIP without infection.

Acute Disease ; Animals ; Ceruletide ; toxicity ; Female ; HSP70 Heat-Shock Proteins ; physiology ; Male ; Mice ; Mice, Inbred C57BL ; Pancreatitis ; etiology ; Systemic Inflammatory Response Syndrome ; etiology ; Toll-Like Receptor 4 ; physiology

BACKGROUNDStudy of the relationship between mast cells and atherosclerosis is mostly dependent on pathological observation and cytology experiments. To investigate the effects of mast cells degranulation on plaque and their possible mechanisms we used apolipoprotein E knockout mice which had been placed perivascular common carotid collar with mast cells degranulator compound 48-80.

METHODSForty apolipoprotein E knockout mice were fed a western-type diet and operated on with placement of perivascular right common carotid collar. Four weeks after surgery, the mice were intraperitoneally injected with compound 48-80 (0.5 mg/kg) or D-Hanks every other day for 4 times. The serum lipids and activity of tryptase were measured. Tissue sections were stained with hematoxylin and eosin. Corresponding sections were stained with toluidine blue and immunohistochemically with antibodies against macrophage-specific antigen, alpha-smooth muscle actin, interleukin-1beta and von Willebrand factor. Simultaneously, basic fibroblast growth factor was detected by in situ hybridization and immunofluorescence.

RESULTSNo pathological change was observed in common carotid non-collar placement but atherogenesis in common carotid collar placement of both groups. There was a significant increase in plaque area ((5.85+/-0.75) x 10(4) vs (0.86+/-0.28) x 10(4) microm(2), P<0.05), the degree of lumen stenosis ((81+/-15)% vs (41+/-12)%, P<0.05), the activity of tryptase in serum ((0.57+/-0.13) U/L vs (0.36+/-0.10) U/L, P<0.05), and the percentage of degranulated mast cells ((80.6+/-17.8)% vs (13.5+/-4.1)%, P<0.05). The expressions of macrophage-specific antigen, alpha-smooth muscle actin, interleukin-1beta, basic fibroblast growth factor and the density of neovessel in plaque were more in the compound 48-80 group than in the control group.

CONCLUSIONSPerivascular common carotid collar placement can promote atherosclerotic plaque formation in apolipoprotein E knockout mice. Compound 48-80 increases plaque area and the degree of lumen stenosis by the mechanism that compound 48-80 promotes proliferation of smooth muscle cells and aggregation of macrophages. Compound 48-80 promotes angiogenesis in plaque. The mechanism is potentially that compound 48-80 increases the expressions of basic fibroblast growth factor mRNA and protein in plaque. Compound 48-80 enhances the expression of interleukin-1beta in plaque.

Animals ; Apolipoproteins E ; genetics ; Atherosclerosis ; chemically induced ; genetics ; metabolism ; pathology ; Carotid Arteries ; drug effects ; pathology ; Fluorescent Antibody Technique ; Immunohistochemistry ; In Situ Hybridization ; In Vitro Techniques ; Male ; Mast Cells ; drug effects ; metabolism ; Mice ; Mice, Knockout ; p-Methoxy-N-methylphenethylamine ; pharmacology

Objective To learn the status of human resources of vaccination clinics in Zhejiang province and to give suggestions for optimizing the allocation.Methods Investigation and the focus group interview were conducted in Zhejiang Province to learn the configuration standards in vaccination clinics.Results The mean number who attended the immunization work every vaccination day in those clinics was five.The number of staff in each clinic differed from county to county but all reached 0.9 persons serving for 10,000 residents.There should not be less than four persons in each vaccination clinic theoretically.It should be necessary to increase one vaccination staff in every 50 injection doses of vaccine workload.Conclusion The staff of vaccination clinics is inadequate and unbalanced in Zhejiang Province.We should increase staff numbers according to workload and served population.

OBJECTIVETo obtain the recombinant rv1837c and rv3803c of Mycobacterium tuberculosis using gene engineering technology and explore their prokaryotic expression, purification, and immunogenicity.

METHODSThe Mycobacterium tuberculosis rv1837c and rv3803c genes were amplified by polymerase chain reaction, and then cloned into the vector pTA2, followed by the subclone into the expression vector pET30a (+). The resulting plasmids, named pET30a (+): rv1837c and pET30a (+): rv3803c, encode recombinant protein containing a hexa-histidine tag on its N-terminus. pET30a (+): rv1837c and pET30a (+): rv3803c were introduced into E. coli BL21 (DE3) by transformation respectively, and the recombinant gene was induced with 0.4 mmol/L isopropyl-D-thiogalactopyranoside. The expressed products were identified by Western blot with hexa-histidine tag antibody and serum from tuberculotic patients. The histidine tagged protein was purified by nickel nitrilotriacetic acid His-Bind resin. Rabbits were immunized with purified recombinant Rv1837c and Rv3803c proteins. Then the purified recombinant Rv1837c and Rv3803c proteins were used to detect antibody in rabbit serum, which had been immunized by Western blot.

RESULTSAfter transformation of the E. coli and induction with 0.4 mmol/L of isopropyl-D-thiogalactopyranoside, recombinant target proteins Rv1837c (relative molecular mass: 92000) and Rv3803c (relative molecular mass: 38 000) were expressed in pET30a (+): rv1837c and pET30a (+): rv3803c system. The expressed protein existed in cytoplasm in an unsoluble form and amounted to 30% and 50% of the total proteins of E. coli. The purity of the purified protein reached 90%. The immunogenicity of the recombinant proteins Rv1837c and Rv3803c was strong, as identified by Western blot.

CONCLUSIONThe prokaryotic expression recombinant plasmids pET30a (+): rv1837c and pET30a (+): rv3803c was successfully constructed and the recombinant proteins Rv1837c and Rv3803c were obtained, which laid a basis for the optimized diagnosis of active tuberculosis.

Antibodies ; metabolism ; Bacterial Proteins ; genetics ; immunology ; metabolism ; Blotting, Western ; Escherichia coli ; metabolism ; Genetic Vectors ; Mycobacterium tuberculosis ; genetics ; immunology ; metabolism ; Plasmids ; metabolism ; Polymerase Chain Reaction ; Recombinant Proteins ; genetics ; metabolism