Venezuelan equine encephalitis (VEE) is a zoonotic disease caused by the Venezuelan equine encephalitis virus (VEEV) complex. This disease has not yet been reported in China, and it is therefore essential to establish a rapid and accurate method for detection of the virus in order to prevent and control this disease. In this study, a one-step real-time quantitative RT-PCR method was developed for the detection of the VEEV complex. A pair of specific primers and a Taqman probe were designed corresponding to a conserved region of the VEEV gene nspl, allowing the detection of all known strains of different sub- types of the virus. Using RNA synthesized by in vitro transcription as template, the sensitivity of this method was measured at 3.27 x 10(2) copies/microL. No signal was generated in response to RNA from Chikungunya virus (CHIKV), nor to RNA encoding the nsp1 fragment of Eastern equine encephalitis virus (EE-EV) or Western equine encephalitis virus (WEEV), all of which belong to the same genus as VEEV. This indicates that the method has excellent specificity. These results show that this one-step real-time quantitative RT-PCR method may provide an effective tool for the detection of VEEV in China.
China ; DNA Primers ; genetics ; Encephalitis Virus, Venezuelan Equine ; classification ; genetics ; isolation & purification ; Encephalomyelitis, Venezuelan Equine ; virology ; Humans ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods

A field survey of three counties in a province in west China analyzed the public health performance from the dimensions of input, output and outcomes, and computed the composite evaluation indexes for public health performance of the sample areas. The indexes so obtained are as follows: Low public health input in general; gap of public health outcomes among various public health programs;generally high outcome performances. The paper proposed to improve the fund raising mechanism for rural public health, build a performance-based investment mechanism, balanced provision of primary public health services, and secure supports from all the society, in an effort to promote public health development in rural areas.

Atherosclerosis is a chronic inflammatory disease,with both the innate and adaptive immune systems responding to many endogenous and exogenous antigens.Subsequent investigations have revealed that an immunomodulatory strategy via active immunization against atherosclerotic plaque antigen(s) could potentially alleviate atherosclerosis.Substantial data from clinical investigations support the key role of immune system in atherosclerosis.So,it may be promising to develop immunotherapy against atherosclerosis.The paper reviews current status of immunization studies and possible associated immunemediated inflammatory mechanisms in atherosclerosis.

Objective To confirm the connotation, constitution and classification of the stakeholders in public hospital. Methods The stakeholders of public hospital were proposed through the brainstorm method and literature search. On this basis, the expert consultation scale was developed by using the score-based approach for reference The stakeholders of public hospital were confirmed and classified through two-round expert consultation. Results The research confirms 16 stakeholders of public hospital on the 80% level of support ratio by experts. There were 10 core stakeholders, 5 latent stakeholders and 1 marginal stakeholder. Conclusion Appropriate stakeholder management strategy should be taken for different types of stakeholders.

Objective To develop a new method for establishing a temperature gradient field in the microchannel on a glass-polydimethylsiloxane ( PDMS ) microfluidic chip and to verify its applicability in the study of cellular thermal biological effect.Methods The establishment and control of the temperature gradient field in the microchannel were implemented by a peripheral indium tin oxide ( ITO) heater and a heating micro-wire embedded in the PDMS chip.The temperature gradient field established in the microchannel was represented by the finite element numerical analysis and temperature-dependent fluorescent dye rhodamine B.Finally, the thermal biological effect, which used cell survival rate of human prostate cancer cells T24 as an indicator, was investigated in the microchannel.Results The results of finite element numerical analysis proved that this method established a temperature gradient field along the length of the microchannel successfully.The distribution range of the temperature gradient field was controlled by the ITO heater, while the gradient of the temperature gradient field was controlled by the heating micro-wire.The measurement result of rhodamine B was identical with the result of the finite element numerical analysis.The thermal biological effect of T24 tumor cell research showed that the cell survival rate decreased with the rise of the regional temperature in the microchannel.Conclusion The method developed in this paper for establishing a temperature gradient field in the microchannel on a glass-PDMS microfluidic chip is simple and easy to implement, and it can be used for parallel study of the cellular thermal biological effect on the microfluidic chip in the future.

Objective To explore the clinical application of blood perfusion quantitatively by contrastenhanced ultrasonography(CEUS) to grade cerebral gliomas during operation. Methods Thirty-one patients with cerebral gliomas were examined by CEUS with Sonoliver software during operation. Maximum intensity (IMAX), time to peak (TTP), rising time (RT) and mean transit time(mTT) of the regions of interest (ROIs) of normal brain tissue and high- and low-grade gliomas were respectively determined and comparatively analyzed. Results Fifteen high-grade gliomas and sixteen low-grade gliomas were postoperatively confirmed by pathologic results. The administration of contrast agent led to higher echo enhancement in cerebral gliomas than normal brain tissues in all cases. The TTP of high- and low-grade gliomas were significantly shorter than that of normal cerebral tissues ( P ＜0.05) ,and the mTT was longer than that of normal brain tissue( P ＜0. 05). There was significant differences of IMAX and TTP between high- and low-grade gliomas(P ＜0.05), but there was no significant difference of RT and mTT between the two groups ( P ＞0.05). Conclusions CEUS with quantitative analysis software on blood perfusion of the tumors can provide valuable information to grade cerebral gliomas.

PurposeThe monitor of organ microcirculation is significant in the diagnosis and treatment of hemorrhagic shock (HS). We established an HS experimental model and evaluated it by contrast-enhanced ultrasound (CEUS), which aimed to evaluate the value of CEUS and time-intensity curve (TIC) in quantitative analysis of renal cortical microcirculation.Materials and Methods The experimental models of HS were established in 30 healthy New Zealand white rabbits by controlled exsanguinations and were divided into four groups according to the shock grade: normal (100% MAP), mild (70% MAP), moderate (50% MAP) and severe (40% MAP). The right kidneys of the experimental model were examined by CEUS. The corresponding parameters of the TIC such as arrival time (AT), time to peak (TTP), peak intensity (PI) and area under the curve (AUC) were measured with the TIC analysis software package when the region of interest was set in superficial of renal cortex.Results The model of HS were successfully established with 30 healthy New Zealand rabbits. Twenty-seven healthy New Zealand rabbits were alive at the end of the experiment, and three died of severe shock. The TIC rose steeply and reached the peak quickly, and then declined slowly to the baseline, which reflected the transition of microbubble in the region of interest. As the hemorrhagic shock model progressed from normal to mild, PI and AUC gradually decreased and the differences were significant (P<0.05); no significant changes were found in AT and TTP (P>0.05). AT and TTP gradually prolonged compared with normal and mild shock groups, and the differences were significant (P<0.05).Conclusion CEUS and TIC can quickly and accurately assess the renal perfusion changes of acute hemorrhagic shock model of animals in a quantitative way. It can be used as a new noninvasive monitoring tool in quantitative analysis of renal cortical microcirculation.

Purpose To study the effects of different doses of ultrasound contrast agent SonoVue upon contrast-enhanced ultrasound (CEUS) in the kidney of healthy rabbits, and to seek the optimal dose of SonoVue. Materials and Methods CEUS was performed in 10 healthy rabbits with GE LOGIQ-E9 by using 8 different doses of SonoVue (ranging from 0.02 ml/kg to 0.16 ml/kg). The quantitative parameters of the time-intensity curve (TIC) were measured and statistically compared. Results The TICs showed that the peak intensity (PI) and the area under curve (AUC) increased with dose when the doses ranged from 0.02 ml/kg to 0.10 ml/kg (r=0.962 and 0.965, P<0.05); when the dose further increased, AUC had little change but PI decreased reversely. The arrival time (AT) shortened along with the increase of the SonoVue dose (r= - 0.917, P<0.05). The dose had a positive correlation with time to peak (r=0.49, P<0.05). Conclusion The parameters of TIC are influenced intensely by different doses of SonoVue. It is important to realize the relationship between SonoVue doses and its effects upon contrast-enhancement ultrasound in microcirculatory quantification. The dose of 0.10 ml/kg appears to be the optimal dose for CEUS in examining kidney of healthy rabbit.

Objective To explore the relationship between the level of serum Klotho with renal impairment in chronic kidney disease (CKD) patients. Methods 100 patients with CKD 1-5 stages (CKD group) and 30 non-CKD patients (control group) were enrolled. The serum of Klotho was detected by ELISA (enzyme-linked immunosorbent assay ) to investigate the correlation between the level of serum Klotho to progression of CKD. Results The levels of serum Klotho in CKD group 9.81 (7.45,12.2) ng/mL and control group 2.97 (1.75,5.23) ng/mL were significantly different (P < 0.01); The Klotho level declined as renal insufficiency progressed. Pearson correlation analysis showed that serum Klotho in CKD patients was positively correlated with eGFR (estimated glomerular filtration rate), blood calcium and hemoglobin,while was negatively correlated with phosphate, age, logserum creatinine, and logFGF-23. Multiple regression analysis showed age, logScr and logFGF-23 were independent factors for expression of serum Klotho in CKD patients. Conclusion Serum Klotho level in CKD patients decreased with the deterioration in renal function and serum Klotho maybe one of indicators to predict the progression of renal function in CKD patients.

Purpose To investigate the value of contrast-enhanced ultrasound (CEUS) in monitoring hepatic parenchymal perfusion quantitatively during resuscitation after hemorrhagic shock (HS).Materials and Methods Forty-five labeled rabbits were randomly divided into five groups.One group was normal control group,and the other four groups were established reversible HS modes by using modified Wiggers method.In the four model groups,one group was chosen as the shock group,and the other three groups received colloid (hydroxyethyl starch) for resuscitation.The three resuscitation groups were named as 1 h group,4 h group,24 h group according to resuscitation time.The arterial blood was collected for testing blood lactic acid (LAC) in each group.The CEUS was performed to form time-intensity curve (TIC) for quantitative analysis.At the end of the experiment,the rabbits were sacrificed and the liver tissues were examined for pathology.Results Compared with the normal control group,the arrival time (AT) and time to peak (TTP) of the shock group were significantly prolonged,and the peak intensity (PI) and area under curve (AUC) of the shock group decreased,all with significant difference (all P＜0.05);in the 1 h resuscitation group,the PI and AUC increased,and the AT decreased,all with significant difference (all P＜0.05);in the 4 h and 24 h resuscitation group,the PI and AUC increased,and the AT and TTP decreased,but all without significant difference (all P＞0.05).Compared with the shock group,the PI and AUC of the 1 h resuscitation group increased,and the AT of 1 h resuscitation group decreased,all with significant difference (all P＜0.05);all the tested parameters of the 4 h and 24 h resuscitation group were significant different (P＜0.05).Conclusion CEUS can quantitatively evaluate the changes of hepatic parenchymal perfusion during HS resuscitation,which has a certain reference for clinical treatment.