Objective To investigate the expression and significance of Peripheral blood of Toll-like receptor-4 and cytotoxic T lymphocyte-associated antigen-4 in patients with essential hypertension. Methods We selected 35 patients with hypertension and 20 healthy people.We used flow cytometry to investigate TLR4 expression levels, and ELISA to detect the expression of CTLA-4. Results TLR4 expression in peripheral blood of patients with hypertension was (8.63 ±1.16)%, significantly higher (5.27 ± 1.25)%.The difference was statistically significant (t = 6.16,P < 0.05); CTLA-4 expression in peripheral blood of patients with hypertension was significantly higher (P<0.05); Hypertensive patients with CTLA-4 positive rate and TC, LDL-C was positively correlated (P<0.05); TLR4 and CTLA-4 was positive correlation (r = 0.886,P < 0.05). Conclusions TLR4 and CTLA-4 were high expression in hypertensive patients with hypertension,and related to hypertension.

Adult ; Female ; Fungi ; Humans ; Mycoses ; Sinusitis ; microbiology

Objective To establish a gene diagnosis assay for spinal muscular atrophy(SMA) in children. Method Analysis of the survival motor neuron (SMN) gene in 19 SMA patients and in 21 normal controls were performed by using polymerase chain reaction - fragment length polymorphism (PCR - RFLP) method. Result Deletion of exon 7and 8 in SMNt gene were found in all 19 SMA patients, while no such changes were found in normal controls. Conclusion The SMNt gene exon 7 and 8 examine can be applied to SMA gene diagnosis, and the PCR- RFLP method have higher sensitivity and particularity to the SMA diagnosis.

The diversity in Tibetan kefir grains and dynamics of the microbial community during the fermentation of Tibetan kefir by PCR-DGGE fingerprinting technique were studied.The results showed that bacterial community of Tibetan kefir grains was more complex than that of yeast.The bacteria communities between different Tibetan kefir grains showed 78%～84% similarity, and yeast 80%～92%.Bands B, E and N of DGGE profiles of the microbial community during fermentation of Tibetan kefir were present throughout the fermentation.Analysis of sequence date showed the majority of the DGGE bands of bacteria community corresponded to LAB, and the most intense band (band E) was completely homology to Lactococcus lactis.

Adult ; Aged ; Cervical Vertebrae ; diagnostic imaging ; Female ; Humans ; Magnetic Resonance Imaging ; instrumentation ; methods ; Male ; Middle Aged ; Radiography ; Spinal Osteophytosis ; diagnosis ; diagnostic imaging ; Young Adult

Background Oxidative damage plays an important role in pathogenesis of age-related macular degeneration( AMD ),and its mechanism is the destroy of blood-retinal barrier.Müller cells is a primary component to stabilize the inner barrier of the blood-retina.Researches showed that epidermal growth factor(EGF) can promote the proliferation and migration of animal Müller cells,but less study was found in the effect of EGF on human Müller cells. Objective The present study was to investigate the effects of EGF on the proliferation and migration of human Müller cells and its molecular mechanism. Methods Human Müller cell line MIO-M1 cells were cultured and incubated,and cultured cells were identified using glial fibrillory acidic protein (GFAP),factor Ⅷ,α-smooth muscle actin( α-SMA ),keratin and S-100.Different concentrations of EGF( 0,1,10,30,100 mg/L)was added in freeserum DMEM,and the positive rate of the cells was calculated using 5-bromo-2-deoxyuridine(BrdU) method.The cells were divided into EGF group,H2 O2 group,EGF + H2 O2 group,glucose oxidase ( GO ) group,GO + EGF group,EGF + LY294002+H2O2 group according to the different intervention,and the effects of LY294002 on the proliferation of Müller cells (A590 )were detected by colorimetric assay for cellular growth and survival( MTT assay).The scratch test of Müller cells was used to assess the influence of EGF(0,1,10,30,100 mg/L)on H2 O2-induced damage of human Müller cell.Western blot was used to detect the cell proliferation under the protection of EGF on co-cultured cells using LY294002 and H2O2 and the activation of Akt signal pathways. Results The proliferative rates of the cells were 28.0％,32.9％,39.0％ in 10,30,100 mg/L EGF groups respectively and obviously higher than those in 0,1 mg/L EGF groups (24.5 ％,26.2 ％ ).Under the H2O2 culture,GO culture,respectively,the A570 value of the Müller cell in high concentrations of EGF groups was significantly increased in comparison with lower concentrations EGF groups with the statistical significance among the groups( F=23.582,P=0.000).Compared with EGF+H2O2 group,the A570value of the Müller cells was lowed in EGF+LY294002+H2O2 group.The maximum migration rate of Müller cells was found in 10 mg/L EGF group.Western blot revealed that the presence of H2O2 reinforced the expression of Akt in Müller cells,however,pretreatment with 100 mg/L EGF antagonized the harmful effect of H2O2 on Müller cells.Meanwhile,pretreatment with EGF and LY294002 reduced the expression of Akt in Müller cells. Conclusions EGF can induce the proliferation and migration of human Müller cells with the strongest effect in 10 mg/L.100 mg/L exogenous EGF has a stronger protection to the Müiller cells against H2O2-induced cell damage by activating the PI3KAkt cell survival pathway.

Objective To investigate the correlation between immune inflammation and overactivity of T helper cells in childhood asthma by cell proliferation assay and activation induced cell death in vitro.Methods Th1/Th2/Th17 cytokines were determined by cytometric bead array.Cell proliferation and activation induced cell death were detected when CD4+ T cells were purified by magnetic beads and stimulated by PHA and antiCD3.At last,mRNA of Fas,FasL and Bcl-2 were mesured by real-time PCR.Results Cytokines of IL-4(2.451± 1.052ng/L vs 1.796 ±0.615 ng/L,P =0.018),IL-10( 1.920 ±0.813ng/L vs 1.390 ±0.162ng/L,P =0.006)and TNF(5.112 ±5.842 ng/L vs 1.506 ±0.551 ng/L,P =0.009) in sera of asthma group were higher than those in control group.Compared to control group,proliferation ability of CD4 + T cells in asthma group was greater ( OD450:0.498 ± 0.052 vs 0.274 ± 0.032,P ＜ 0.001 ) and apoptosis rate was lower( 35.62 ± 0.05 ％ vs 65.28±3.85％,P ＜0.001 ).mRNA expression of Fas in asthma group was lower but Bcl-2 was higher than those in control group.Conclusion It is implicated that defective expression of Fas and over expression of Bcl-2 in CD4+ T cells may contribute to apoptosis inhibition and cell proliferation,which could explain overeactivity of CD4 + T cells and lvmphocvte infiltration in childhood asthma.

Objective To evaluate the outcome of super-selective embolization of renal artery for severe hemorrhage after percutaneous nephrolithotomy (PCNL) and its effect on renal function. Methods From May 2008 to Feb 2010,severe bleeding occurred in 7 patients after PCNL in our hospital.(5 males and 2 females,average age of 54.9 years ).All cases were treated with super-selective renal angiography and 6 cases underwent microcoil embolization. Results Renal angiography showed pseudoaneurysm in 5 cases,pseudoaneurysm with arteriovenous fistula in 1 case and no severe bleeding in 1 case.Successful coil embolization was confirmed in 6 cases by angiography,and bleeding stopped within 3 -7 days after embolization.Serum creatinine and blood urea nitrogen were 59 -98 μmol/L(mean,78.3 μmol/L) and 1.86 -6.92 mmol/L( mean,4.8 mmol/L) 2 weeks after embolization,respectively. Conclusions Super-selective embolization of renal artery for severe hemorrhage after percutaneous nephrolithotomy has the advantages of remarkable hemostatic effects and mild impaired renal function,which is of the first choice.

Objective To analyze the intratumoral and peritumoral microvessel density (MVD) and microlymphatic vessel density (MLVD) in pancreatic cancer and record the expression of vascular endothelial growth factor(VEGF)-C and VEGF-D. And to explore the significance of VEGF-C and VEGF-D during the lymphatic metastasis and development of pancreatic cancer. Methods The expression of VEGF-C and VEGF-D, VEGF-R3, CD34 were assayed by immunohistochemical staining in 30 cases of pancreatic cancer tissues. Results The positive rates of VEGF-C and VEGF-D protein in the central portion of tumors (30% and 16.7%) were significantly lower than those in the marginal portion (73.3% and 56.7%), P <0.01. The group with high expression of VEGF-C and VEGF-D in the marginal portion had significantly higher incidences of lymph node metastasis, lymphatic invasion and venous invasion( P <0. 01 ). MLVD in both of the VEGF-C and VEGF-D positive groups was higher than that in the negative groups( P <0. 01 ), and the lymph node me-tastasis increased. MVD in VEGF-C positive group was significantly higher than that in the negative group. MVD had no significant difference between VEGF-D positive and negative group ( P =0. 07). Conclusions The expression of VEGF-C and VEGF-D in the marginal portion of tumor is significantly correlated with lym-phatic metastasis in pancreatic cancer patients, and may induce lymphangiogenesis. VEGF-C may play an im-portant role in the regulation of angiogenesis and lymphangiogenesis in pancreatic cancer, and VEGF- D maybe only participate in the regulation of lymphangiogenesis.

Objective To study the expression of secondary lymphoid-tissue chemokine (SLC)、 CCR7 and its correlation with clinical pathology and lymphangiogenesis in pancreatic adenocarcinoma (PAC). Methods The tissue specimens including PAC, the cancerous peripheral tissues, the normal pancreatic tissues and peripheral lymph nodes were obtained from 30 patients with PAC. The expressions of SLC and CCR7 in these tissues were assayed by immunohistochemical staining and reverse transcription polymerase chain reaction (RT-PCR). MIND marked by VEGFR-3 was detected by morphometric analysis, and the relationship between MLND and clinical pathology of PAC was analyzed. Results In all the specimens, the positive rates of SLC protein in PAC, the cancerous peripheral tissues, the normal pancreatic tissues and peripheral lymph nodes were respectively 16. 7%, 43. 3%, 76. 7% and 46. 6%. The positive rates of CCR7 protein in PAC, the cancerous peripheral tissues, the normal pancreatic tissues and peripheral lymph nodes were respectively 76. 7%, 66. 7%, 30. 0% and 70. 0%. The results of RT-PCR and fluorescence quantitative real-time PCR indicated that the expression levels of CCR7 mRNA in PAC tissues, the cancerous peripheral tissues and peripheral lymph nodes were higher than that in the normal pancreatic tissues ( P <0. 01 ). There was no significant correlation between the expression of SLC protein with MLVD of PAC ( P > 0. 05 ). There was 23 specimens that the CCR7 protein was positive, and among these specimens the MIND was higher than that in negative group of CCR7 protein (P = 0.004). Conclusions The expression of SLC was not related to lymphatic metastasis and TNM stages of PAC. The expression of CCR7 was significantly associated with lymphatic metastasis and TNM stages of PAC, and the high expression of CCR7 in PAC tissues was significantly associated with lymphangiogenesis of PAC.