AIM: To study the effects of tumor necrosis factor-α (TNF-α) on the transcriptional activity of nuclear factor-erythroid 2-related factor 2 (Nrf2) in pulmonary microvascular endothelial cells. METHODS: Rat pulmonary micro-vascular endothelial cells (PMVECs) were cultured by lung tissue block pasted methods, and identified immunocytochemically using Ⅷ factor-related antigen. The cells were treated with different doses TNF-α (prepared in serum-free medium) for 4 h. Subcellular localization and levels of Nrf2 in PMVECs were observed with immunocytochemical methods. Nuclear extract were obtained to assayed transcriptional activity of Nrf2 with EMSA. Total RNA were isolated to assay the mRNA expression of Nrf2 by RT-PCR. RESULTS: The protein level of Nrf2 in the nuclei and transcriptional activity increased dose-dependently in PMVECs after treated with TNF-α at concentrations of 2.5, 5.0 or 10.0 μg/L. However, the protein level of Nrf2 in nuclei and transcriptional activity decreased dose-dependently in PMVECs after treated with TNF-α at concentrations of 20 or 40 μg/L. No different mRNA expression of Nrf2 in PMVECs treated with TNF-α at all concentration above was observed. CONCLUSION: Transcriptional activity of Nrf2 increases in PMVECs treated with low or moderate doses of TNF-α and decreases in PMVECs treated with high doses of TNF-α.

Objective: To examine the influencing factors of the efficiency of county-level centers for disease control and prevention ( CDCs) in China. Methods:458 county-level CDCs were selected based on a systematic sam-pling method. Multilevel modeling was used to analyze the region-level and institution-level influencing factors affect-ing the efficiency of CDCs. Results: It was found that the region ( province) is associated with the efficiency of a CDC. The region-level factor of population density exhibited a significant influence, while the institution-level factors such as the proportion of health technicians, service income and CDC laboratories per capita also had an influence on overall efficiency. Conclusions: Both the region-level and institution-level determinants influence efficiency. Multi-level modeling can help researchers gain a comprehensive understanding of the influencing factors that affect the CDC efficiency.

Objective To explore the survival/proliferation,apoptotic and death effects of keratinocyte growth factor (KGF) in alveolar epithelial type Ⅱ cells (AT Ⅱ Cs) exposed to hyperoxia.Methods Primary culture of AT Ⅱ Cs from the Sprague-Dawley rat fetuses was studied under room air condition (210 mL/L O2) and hyperoxic condition (950 mL/L O2) for 0.5-12.0 h.Various concentrations of KGF (15 μg/L,25 μg/L,50 μg/L,75 μg/L,100 μg/L)were added into the cell cultures.Cells were randomly divided into room-air group,room-air-KGF group,hyperoxic-exposure group and hyperoxic-exposure-KGF group.The levels of intracellular reactive oxygen species (ROS),cleaved cysteinyl aspartate specific proteinase-3 (Caspase-3),cell death and proliferation of AT Ⅱ Cs were measured by flow cytometer,Western Blot,release of lactate dehydrogenase assays (LDH assays) and 3-(4,5-Dimethyhhiazol-2-yl)-2,5-diphenyhetrazolium bromide assays (MTT assays),respectively.Results Under room air condition,KGF could significantly increase AT Ⅱ Cs proliferation with 15-100 μg/L in a dose-dependent manner and significantly decrease LDH production at concentrations of 25-100 μg/L.Exposure to hyperoxia resulted in a significant increase in intracellular ROS production in AT Ⅱ Cs in a time-dependent manner compared with that of the room air group.Cell viability decreased and LDH release increased significantly in a time-dependent manner when AT Ⅱ Cs were exposed to 950 mL/L O2 for more than 4 h.After exposure to hyperoxia for 0.5 h and 1 h,KGF could significantly increase AT Ⅱ Cs proliferation in 15-75 μ g/L and significantly decrease LDH production at concentrations of 25-75 μg/L.After exposure to hyperoxia up to 4 h,higher viability was observed in 15 μg/L and 25 μg/L KGF group,and lower death rate presented in 25-100 μg/L KGF group.Further,prolnged hyperoxic exposure for 8 h,high viabilitv was shown only in 50 μg/L KGF group,and less death rate was observed only in 75 μg/L KGF group.In addition,no significant difference in viability and mortality was found between hyperoxic group and hyperoxic-KGF group after hyperoxic exposure for 12 h.Expression of cleaved Caspase-3 was significant higher after 4 h and 8 h hyperoxic exposure than that in room-air group ;at the same time,by adding 25 μg/L and 75.μg/L KGF led to decreased expression of Caspase-3 was detected,compared to hyperoxic group.Conclusions KGF may promote survival/proliferation,inhibited apoptosis and death of rat fetal AT Ⅱ Cs in room air condition or under temporary exposure to hyperoxia in vitro.However,prolonged exposure to hyperoxia may decrease the sensitivity of AEC Ⅱ Cs to KGF and limit its protective effects on lung injury.

BACKGROUND:Currently, literatures about autologous vein transplantation are few, and the research on the effect of different parts of autologous vein transplantation are not found yet. OBJECTIVE:To summarize the experiences of establishing the fistula using autologous vein transplantation so as to investigate the method of improving the success rate of surgery. METHODS:We analyzed retrospectively the data of 40 cases of establishing the fistula using autologous vein transplantation, and then compared the successful rate of autologous vein transplantation fistula, blood flow and operating time, thereby analyzing the influence of diabetes melitus on the successful rate of autologous vein transplantation fistula. RESULTS AND CONCLUSION:The successful rates of autologous vein transplantation fistula at different parts ranging from high to low were as folows: the cephalic vein, great saphenous vein, basilic vein and smal saphenous vein. Blood flow of the upper limb for vein transplantation fistula was obviously higher than that of the lower limb (P < 0.05). The operating time of autologous vein transplantation fistula was longer in the upper limbs than in the lower limbs (P < 0.01). For patients with diabetes melitus, the successful rate of autologous vein transplantation was markedly lower than those with no diabetes melitus (P < 0.01). For the hemodialysis patients with poor upper limb superficial vein, autologous vein transplantation is a better way of establishing the vascular access. Vein transplantation of the upper limbs is better than that of the lower limbs in success rate and operating time. Autologous vein transplantation fistula is not suitable for the patients with diabetes melitus.

Objective Using an adenoviral vector , the wild-type PTEN gene was transduced into activated hepatic stellate cell (HSC) in vitro and the phosphorylation status of Akt were investigated. Methods The wild type PTEN gene was transduced into activated HSC in vitro mediated by adenoviral vector. The expressions of PTEN and total Akt in HSC were measured by Western blot and Real-time fluorescent quantitation PCR. And the expressions of phosphorylated Akt (Thr308) in HSC was determined by Western blot. Results The data showed that exogenous wild type PTEN gene was successfully transduced and expressed in activated HSC in vitro. The over-expression of wild type PTEN resulted in the significant down-regulated expression of phosphorylated Akt (Thr308) in activated HSC (P < 0.01). But no significant defferences were found in the expression of total Akt in activated HSC at both transcriptional and translational levels(P>0.50). Conclusions The overexpression of wild-type PTEN can negatively regulate PI3K/Akt signaling transduction by inhibiting the phosphorylation of Akt in activated HSC in vitro.

BACKGROUND:Previous studies have shown that puerarin can inhibit the differentiation of osteoclasts,and the expression of Notch signaling pathway-related proteins such as Notch1,HES1,and Jagged1 is decreased.However,the specific mechanism of the Notch1 signaling pathway for the inhibition of osteoclast differentiation by puerarin is not clear. OBJECTIVE:To explore the effect of Notch signaling pathway on puerarin inhibiting the differentiation of mouse macrophage Raw264.7 into osteoclasts. METHODS:Raw264.7 cells were divided into seven groups for intervention culture.Blank control group was cultured in high-sugar DMEM medium;the osteoclast induction group was cultured in osteoclast induction medium;the puerarin intervention group was cultured with 50 μmol/L puerarin at the same time of osteoclast induction;Notch1 siRNA control group,Notch1 siRNA group,Notch1 overexpression control group and Notch1 overexpression group were transfected with Notch1 siRNA control sequence,Notch1 siRNA,Notch1 overexpression control plasmid and Notch1 overexpression plasmid,respectively,and then cultured with osteoclast induction medium and puerarin.The number and size of osteoclasts were observed by tartrate-resistant acid phosphatase staining,the skeleton formation of osteoclasts was observed by F-actin staining,and the gene expression level of osteoclast formation markers was detected by RT-PCR. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining results showed that puerarin intervention could inhibit the generation of osteoclasts,Notch1 silencing could further reduce the number of osteoclasts,while the number of osteoclasts in the osteoclast-induced group increased significantly after Notch1 overexpression.The results of F-actin showed that Raw264.7 cells could form a well-defined F-actin ring after osteoclast induction.Puerarin intervention would inhibit the formation of cytoskeleton,and Notch1 silencing could aggravate the inhibitory effect of cytoskeleton formation,while Notch1 overexpression could alleviate this inhibitory effect of puerarin.RT-PCR results showed that puerarin could inhibit the mRNA expression levels of tartrate-resistant acid phosphatase,Cathepsin K and c-Fos,the expression of the above-mentioned three factors decreased significantly after Notch1 gene silencing,and Notch1 overexpression could upregulate the expression of these factors.These finding indicate that puerarin inhibits the differentiation of Raw264.7 cells into osteoclasts through the Notch signaling pathway.