We aimed to develop a real-time polymerase chain reaction (PCR) detection method for the Reston subtype of the Ebola virus. The NP gene of the Reston subtype of the Ebola virus was selected as the detection object. Sequences of different subtypes of Ebola viruses were aligned using Clustal W software. The most unique and conserved regions of the Reston subtype of the Ebola virus were recruited as candidate sequences for specific primers. Primer Express and Primer Premier 5. 0 software were used to filter the optimal pair of primers for detection. Real-time PCR was carried out using optimized parameters and positive DNA prepared by serial (tenfold) dilution of a recombinant plasmid and by plotting a standard curve. In addition, the reproducibility, accuracy, and specificity of the assay were tested. Results showed that the sensitivity of detection of the Reston subtype of the Ebola virus by real-time PCR could reached 10(2) copies/microL. The linear relationship (R2) reached 0.997, the slope of the standard curve was -0.3101, and amplification efficiency was 110.145%. A sharp and narrow melting peak appeared at 79.94 degrees C for all standards in different dilutions. In conclusion, a fast and sensitive real-time PCR detection system for the Reston subtype of the Ebola virus was developed. This system could be used as a supplementary diagnostic and monitoring approach for basic and clinical studies on the Reston subtype of the Ebola virus. The detection system does not require expensive technology or specialist operators.
DNA Primers ; genetics ; Ebolavirus ; classification ; genetics ; isolation & purification ; Hemorrhagic Fever, Ebola ; virology ; Humans ; Real-Time Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Objective To determine the capability of echocardiography to identify primary cardiac valve myxomatous degeneration (PCVMD) compared to pathological findings and to determine the echo features of PCVMD. Methods Echocardiograms were retrospectively compared with pathological findings of 1080 patients who underwent surgery for moderate or severe valve regurgitation. PCVMD of the mitral,aortic and tricuspid valves was retrospectively identified, with a comparison of the echocardiography and pathology findings,to calculate the prevalence of PCVMD,and to summarize its echocardiography features.Results A total of 104 patients were diagnosed with PCVMD (prevalence of 9. 62%) with 117 lesions.Valvular regurgitation were confirmed by echocardiography in all patients( 100 % ). Valve morphology change including valve prolapsed, valve thickening and redundancy were confirmed by echocardiography in 101 lesions(86.3 % ), but suggestive diagnosis were done by echocardiography in only two patients. Conclusions In patients undergoing surgery for valvular regurgitation,a high prevalence of PCVMD was found. PCVMD had distinctive echocardiographic features,suggesting its preoperative diagnosis.

Objective: To investigate the effect of bone marrow mesenchymal stem cells (MSC) on the variation of intestinal permeability damaged by superior mesenteric artery ischemia and reperfusion. Methods: Bone marrow mesenchymal stem cells were isolated from cavity of tibias and femurs of male Sprague Dawley rat in a sterile condition, and were cultured and proliferated in plastic dishes. 10 week old female Sprague Dawley rats were randomly divided into three groups:group A (sham group), group B (MSC group) and group C (saline group). In group B and group C, the superior mesenteric artery (SMA) of the animals were seperated and occluded by non-invasive vascular clamp for 45 minutes. Immediately after removing the vascular clamp,1×10~7 MSC suspended in 0.5 ml sterile L-DMEM and the same volume of normal saline was submucosally injected into the small intestine at ten different points in group B and group C, respectively. In group A, the animals were only underwent laparotomy without clamping the SMA. 3 days and 6 days after the operation, 100 mg lactulose and 50mg mannitol dissolved in 2 ml distilled water were administrated by oral gavage and urine during 6 h experiment was collected for assaying the L/M ratio before sacrificing the animals. The donor derived MSC was identified by Y chromosome in situ hybridization in ileum tissue, and the serum D-lactate level was determined. Results: The donor derived MSC could home to the ischemia/reperfusion injured intestinal mucosa, and the intestinal permeability was much lower in group B (MSC group) than that in group C (saline group)(P<0.05). Conclusion: Mesenchymal stem cells can reduce the small intestinal mucosal permeability impaired by ischemia/reperfusion, and can participate in the preservation of integrity of the damaged gut mucosal mechanical barrier.

Objective To establish a method for determinning quetiapine in human hair by HPLC.Methods A reverse phase HPLC system was performed on Inertsil ODS-C18 column (4.6 mm ×150 mm,5μm)with the mobile phase consisted of 0.01 mol/Lammonioum-methanol(32︰68).The flow rate was 1.0 mL/min,column temperature was 40℃,the detection wavelength was 254 nm.N-hexane was used as extracting solvent.Results The calibration curve was linear in the range of 5-200 ng/mg.for quetiapine.The recovery of extraction >75.0%, the recovery of method was between 95.0% and 105.0%, the intra-day and inter-day precision were all no more than 10.0%.This method met the requirements of biological samples. Conclusion A HPLC method of concentration of quetiapine in human hair is established, which is simple,sensitive,accurate and has a certain value.

Panax notoginseng (BurK.) F.H.Chen is a traditional Chinese medicinal material with a time-honored history of cultivation.There are a series of problems,such as high pesticide residues,serious disease and pest,and continuous cropping obstacles in the process of the cultivation of notoginseng.Pollution-free cultivation is an effective strategy for the sustainable development of notoginseng industry.We herein summarized three points of the pollution-free cultivation of notoginseng in this review.The standard of lands suitable for the cultivation of notoginseng was established on the basis of the analysis of medicinal plants around global producing areas.The integrated measures of soil improvement were put forward by cfficient rotation and soil disinfection with new varieties breeding combined with the management of water,light and fertilization,and the safe and low-toxic methods of disease and pest control.Additionally,the mode of wild tending should be carried out when the marker-assisted breeding of new varieties was developed,and the platform of comprehensive disease and pest control was founded.Above-mentioned points can effectively perfect and optimize the pollution-free cultivation of notoginseng and promote sustainable development of notoginseng industry.

Objective To express virus-like particles(VLP) of enterovirus 71 (EV71) in Han-senula polymorpha.Methods The coding sequences of P1 and 3CD genes of EV71 were optimized accord-ing to codon usage bias of Hansenula polymorpha for achieving high expression , and then cloned into the ex-pression vector PMV of Hansenula polymorpha .The recombinant expression vector PMV-P1-3CD was trans-formed into Hansenula polymorpha AU 0501 .The transformants were stably cultured in selective medium (Yeast Nitrogen Base) and screened for strains with positive P1 and 3CD genes by PCR.Then an induced cultivation on the recombinant strains were performed in a medium supplemented with methanol to a final concentration of 1.0%and the expressed products were analyzed by SDS-PAGE and Western blot assays to select high expression strains .The high expression strains were cultured in 30 L fermentor and its fermenta-tion products were analyzed by electronic microscope after purification .Results EV71 recombinant expres-sion strains were successfully constructed .The results of SDS-PAGE showed that the expressed products had obvious VP3, VP1, VP0 protein bands with molecular weights of 26×103, 33×103 and 35×103, respective-ly, which were consistent with the expected molecular weight of the fusion proteins .Western blot demonstra-ted that the expressed products could be specifically recognized by the polyclonal antibody against EV 71-VP1 at 33 ×10 3 , indicating its high immunoreactivity .ELISA confirmed that the expression level of EV 71 fermen-tation products was reached to 200 mg/L.Electronic microscope analysis showed that the VLP of recombi-nant EV71 were 24-30 nm in diameter with normal structure .Conclusion The virus-like particles of human enterovirus 71 are successfully expressed in Hansenula polymorpha , which provides a foundation for the fur-ther development of EV 71 VLP vaccine .

Objective To investigate effects of antioxidant stress protein hem e oxygenase-1 (HO-1) on lipopolysaccharide (LPS)-induced endoplasm ic reticulum stress (ERS) of rat hepatocytes. Methods The BRL cells (rat hepatocyte cell line) were cultured. The hepatocytes were treated with LPS, LPS+HO-1 si RNA , HO-1 siRNA and PB S solution, respectively. The cell viability was m easured by trypan blue ex-clusion test. The apoptosis cells were detected by the fluorescent dye Hoechst 33258. E xpressions of GR P78, C HO P, caspase-12 and HO-1 were detected by Western blotting. Results LPS caused an in-crease of HO-1 protein expression of rat hepatocytes in a dose-dependent and tim e-dependent m anner, a up-regulation of GRP78, CHO P and caspase-12, a decrease in cellviability,and an increase in apopto-sis rate of hepatocytes. Pretreatm ent of HO-1 siRNA inhibited the up-regulation of LPS-induced HO-1, however, aggravated ERS and cellular injury. Conclusion HO-1 inhibites ERS-m ediated cellular injury of rat hepatocytes induced by LPS.

Non-coding RNA could regulate gene expression, involved in epigenetic modification, and participate in the cell differentiation, proliferation, apoptosis, and other life activities. Noncoding RNA also plays a crucial role in cancer occurrence, cancer cell invasion and distant metastases. Through mediating genome hyper-methylation, transcriptional regulation, regulation of transposable sequences, maintenance of genomic imprinting and DNA damage repair, noncoding RNA could regulate the growth and apoptosis of pancreatic cancer cells. Understanding the molecular mechanism of non-coding RNA in the development process of pancreatic cancer has important theoretical and practical value in the diagnosis, treatment and prognosis.
Apoptosis ; Cell Differentiation ; Epigenesis, Genetic ; Humans ; Pancreatic Neoplasms ; genetics ; Prognosis ; RNA, Untranslated

Objective , To investigate the influence of blood pressure variability on cerebral infarction in older men. Methods Ambulatory blood pressure was measured in 1527 elderly men ( older than 65 yrs) with atherosclerosis. All cases were divided into 2 groups: Six hundred and seven patients with cerebral infarction ( group A)and 920 patients without cerebral infarction ( group B). Smooth curve method was used to analyze each patient's ambulatory blood pressure data and the trend of each patient's blood pressure curve was portrayed. The differences between the actual blood pressure and the blood pressure on the curve was defined as blood pressure variability,and the blood pressure variability between the 2 groups was compared. Results The systolic blood pressure variability in 24 hours in group A was significantly higher than that in group B( [8.4'±2. 2]mm Hg vs [ 8.0 ± 2. 0 ] mm Hg, P ＜ 0. 01 ), especially for the systolic blood pressure variability in daytime( [ 8. 2 ± 2. 2 ] mm Hg vs [ 7. 8 ± 2. 1 ] mm Hg, P ＜ 0. 01 ). However, the systolic blood pressure variability at night was not significantly different between the 2 groups( [ 8.9 ± 3. 9 ] mm Hg vs [ 8. 7 ± 3.7 ] mm Hg,P ＞ 0. 05 ). There were no significant difference between the diastolic blood pressure of 24 hours( [5. 5 ± 3.8 ] mm Hg vs [5.5 ± 1.5 ]mm Hg,P ＞0. 05),during daytime([5.4 ± 1.5]mm Hg vs [5.3 ± 1.4] mm Hg,P ＞0.05)and nighttime ( [ 6. 1 ± 2.7 ] mm Hg vs [ 6. 1 ± 2. 6 ] mm Hg, P ＞ 0. 05 ). Conclusion In elderly men with atherosclerosis,cerebral infarction was closely related to systolic blood pressure variability,but independent of nighttime systolic blood pressure and diastolic blood pressure variability.

Objective To explore the important anatomic structures associated with pulmonary veins and similar sections to transthoraeic echocardiography using 64一slice spiral CT and tO locate the pulmonary veins on echocardiography. Methods The transthoracic echocardiography and 64-slice spiral CT were performed in 12 patients with atrial fibrillation.The 3-dimensional information of four pulmonary veins on CT was analyzed and the relationship between the locations of four pulmonary veins and adjacent anatomic structures were observed.The comparison of CT and echocardiography for assessing the location of pulmonary veins was performed.Results All of the pulmonary veins were demonstrated optimally without abnormal structures and anatomic variations.The ostium of right superior pulmonary vein adjoined the atrial septum and the proximal of the right superior pulmonary vein was near to the superior vena cava.The right inferior superior pulmonary was a little bit far from atrial septum.The left superior pulmonary vein adjoined the left atrial auricle.The left inferior pulmonary vein was a little bit far from the 1eft atrial auricle and adjoined the thoracic descending aorta.According to adjusting the angle of the probe on echocardiography,the four pulmonary veins could be demonstrated in the standard parasternal left ventricular long-axis view, parasternal short-axis view and apical four chamber view.The non-standard views assisted in identifying the pulmonary veins.Conclusions Transthoracic echocardiography can demonstrate all the four pulmonary veins and also locate the spatial location based on the adjacent anatomic structures.