Objective To optimize the PCR primer sets for Simian virus 40 (SV40) detection and establish an assay method for SV40 which is of high sensitivity, strong specificity, broad applicability. Methods Two pairs of PCR primers were designed of based on 21 different SV40 strains genome by Primer Premier 5.00 software, and the features of two pairs of PCR primers were analyzed by Oligo software (version 6.71), conservative nucleotide of two pairs of PCR primers and the PCR amplification product were analyzed by DNAMAN software (version 6.0.40). Two pairs of new-built PCR primers were compared with those derived from China pharmacopoeia (Clip) in these aspects. The detection sensitivity of four pairs of PCR primers were analyzed using different SV40 DNA diluent as PCR template. The detection specificity of four pairs of PCR primers were analyzed using sterile water, Vero cell DNA, SV40 DNA as PCR template, respectively. Results The sequences of the new PCR primer sets VP1 and T are conservative for 21 Strains. The sequences of PCR primer sets GCVP1 and GCT are conservative for SV40 strains whose accession No. is J02400, NC_001669, AF316139 and AF316141. As far as the same diluent SV40 DNA template is concerned, the PCR amplification efficiency of PCR primer set VP1 and T is higher than that of PCR primer set GCVP1 and GCT. There are non-specific band in nucleic acid electrophoresis for amplification products of PCR primer sets GCVP1 and GCT, whereas there are no non-specific band in nucleic acid electrophoresis for amplification products of PCR primer sets VP1 and T. Conclusion The new assay method for SV40 nucleic acid sequence has many better qualities than those in Chp such as high sensitivity, strong specificity, broad applicability, conservation of primers and their amplification products and so on.

Objective: To establish an HPLC method for the simultaneous determination of two ingredients in indometacin and furazolidone suppositories.Methods: The analysis was performed on an XTerra(R) RP18 column (250 mm×4.6 mm, 5 μm) with gradient elution of methanol and 0.01 mol·L-1 potassium dihydrogen phosphate-triethylamine (100∶0.02) at the flow rate of 1.0 ml·min-1.The column temperature was 30 ℃ and the detection wavelength was 263 nm.The injection volume was 10 μl.Results: The peaks of furazolidone and indometacin were successfully separated.The linear range of calibration curves was 5.12-81.87 μg·ml-1 (r =1.0000) and 3.78-60.45 μg·ml-1 (r =1.0000), respectively.The average recovey was 99.6% (RSD =1.5%, n =6) and 100.3% (RSD =1.6%, n =6), respectively.The limit of quantification (LOQ) was 0.02 μg·ml-1 and 0.05 μg·ml-1, respectively.Conclusion: The established method is validated to be suitable for the quality control of indometacin and furazolidone suppositories.

Protamines are a group of highly basic proteins first discovered in spermatozoon that allow for denser packaging of DNA than histones and will result in down-regulation of gene transcription[1].It is well recognized that the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) encodes P6.9,a protamine-like protein that forms the viral subnucleosome through binding to the viral genome[29].Previous research demonstrates that P6.9 is essential for viral nucleocapsid assembly,while it has no influence on viral genome replication[31].In the present study,the role of P6.9 in viral gene transcription regulation is characterized.In contrast to protamines or other protamine-like proteins that usually down-regulate gene transcription,P6.9 appears to up-regulate viral gene transcription at 12-24 hours post infection (hpi),whereas it is non-essential for the basal level of viral gene transcription.Fluorescence microscopy reveals the P6.9's co-localization with DNA is temporally and spatially synchronized with P6.9's impact on viral gene transcription,indicating the P6.9-DNA association contributes to transcription regulation.Chromatin fractionation assay further reveals an unexpected co-existence of P6.9 and host RNA polymerase Ⅱ in the same transcriptionally active chromatin fraction at 24 hpi,which may probably contribute to viral gene transcription up-regulation in the late infection phase.

Objective To explore the sleep characteristics of submariners during a long-term voyage, so as to provide scientific evidence for ensuring submariners with good sleep during long-term voyages. Methods The sleep status of submariners who participated in a long-term voyage was tested by Self-Rating Scale of Sleep (SRSS) before the voyage, and before and after each voyage section during the voyage. The sleep status variation of submariners who performed different types of tasks, from the beginning to the end of each voyage section and of each resting-on-the-sea section was analyzed respectively. Comparison of sleep scores was performed between submariners and surface ship crew in the second voyage section. Numbers of submariners with sleep problem were compared in each voyage section. Results Generally speaking, submariners' sleep status at the end of voyage section was significantly worse than that at the beginning of voyage section and that before the whole voyage (P<0.001, P<0.01), and the sleep status at the beginning of the third voyage section was significantly worse than that before the whole voyage (P<0.05). Submariners had a steady sleep status when taking a resting-on-the-sea before starting their first voyage section, which was no significant difference from that before the whole voyage (P>0.05). After finishing a voyage section and taking a resting-on-the-sea, submariners' sleep status returned to the level of pre-voyage (P>0.05), and was significantly better than that before the resting-on-the-sea (P<0.05, P<0.01). After finishing two voyage sections and then taking a resting-on-the-sea, the submariners' sleep status showed no obvious variation (P>0.05). Compared with that of surface ship crew who accomplished the same voyage section, submariners had an obviously better sleep status after taking a resting-on-the-sea (P<0.05). Meanwhile, submariners who finished a voyage section showed a significantly worse sleep status than those resting on the sea (P<0.01) and surface ship crew who finished a same voyage section (P<0.05). In each voyage section, submariners with sleep problems who finished resting-on-the-sea were significantly less than those who finished navigation (P<0.001, P<0.05). There was no significant difference in the number of submariners with sleep problems between those who taking non-resting and taking resting-at-dock after finishing the first voyage section (P>0.05), but the latter was significantly more than the former when the second voyage section was finished (P<0.05). During the resting-on-the-sea period, the numbers of submariners with sleep problems in both the second and the third voyage section were significantly more than those in the first voyage section (P<0.05, P<0.01). The numbers of submariners with sleep problems who implemented the third voyage section were significantly more than those who implemented the first and the second voyage section (P<0.01). Conclusions Generally, the sleep quality of submariners is significantly worse after accomplished a voyage section task, and the degree of sleep problems may be accumulated to worse and worse along with the increase of long-term voyage time. Whereas, submariners may have a significantly better sleep status after taking a resting-on-the-sea, implying that resting-on-the-sea is an effective way to ensure submariners a good sleep during a long-term voyage.

Objective To investigate the effects of medium-and long-chain fatty acid triacylglycerols on blood lipid in hyperltriacylglycerolemic subjects. Method A case-control clinical trial was carried out,in which 112 subjects with hypertriacylglycerolemia were enrolled and divided into two groups. These two groups were randomized to long-chain fatty acid triacylglycerol oil(LCT) ,and medium-and long-chain fatty acid triacylglycerol oil(MLCT) . All subjects were requested to ingest fixed energy intake and perform physical activity daily during the whole study. Both of the oils were asked to consume at 25-30g daily for consecutive 8w. Anthropometric measurements and blood biochemical variables were measured at the initial and final time of the study. Results Eleven subjects were excluded from the study because of various reasons. There were 50 and 51 subjects left in LCT and MLCT group,respectively. No difference of sex distribution was noted between two groups. There were also no significant differences in daily intake of energy,protein,fat and carbohydrate,as well as daily physical activity time between two groups at 0,2,4,6 and 8w. The subjects consuming MLCT demonstrated significant decreasesin body weight,BMI,WC,HC,WHR,body fat,body fat % compared with the initial values after 8w. A greater extent of decrease in body weight,BMI,WC,body fat and body fat% was found in MLCT group than in LCT group. The levels of TG,ApoB,ApoAII,ApoC2,ApoC3 in MLCT group were significantly lower than those in LCT group after 8 w,and the extent of decrease in these indicators was much greater in the former than in the latter. Conclusion Consumption of medium-and long-chain fatty acid oil may help to reduce body weight,body fat and concentration of blood triacylglycerol and improve apolipoprotein metabolism in hypertriacylgly cerolemic subjects under an appropriate dietary regime.

Objective To compare the effects of oil with medium-long-chain triacylglycerols (MLCT) on lipid metabolism in hypertriglyceridemic patients with different sex. Method One hundred and one subjects with hypertriglyceridemia were enrolled and divided into two groups. One group consumed oil with MLCT (50 subjects) and another group with long-chain fatty acid triglyceride LCT (51 subjects) for 8 w. No sex difference was noted between two groups. All subjects were informed to consume 25-30g oils daily with fixed energy and perform exercise informed. Anthropometric measurements of body weight,waist circumference (WC) body fat weight, total visceral and subcutaneous fat area in abdomen and blood biochemical variables of aspartate amino- transferase (AST), alanine aminotransferase (ALT), glucose, total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein A1 (ApoA1), ApoB, ApoAⅡ, ApoC2, ApoC3 and ApoE were measured at initial and final time of the experiment. Results The decreases in body weight, BMI, WC, body fat weight, total and subcutaneous fat area, and blood TG, LDL-C, ApoAⅡ, ApoB, ApoC2, ApoC3, ApoE in male subjects consuming MLCT were much greater than those consuming LCT after 8 w. However, there were no significant differences in these parameters in female subjects between two groups. Conclusion Consumption of oil with MLCT reduces body weight, body fat and blood TG and improves apolipoprotein metabolism in male hypertriacylglycerolemic subjects, but not in female.

ObjectiveTo evaluate the clinical effectiveness of posterolateral autograft bone graft fusion transpedicular screw system internal fixation and anterior debridement in the treatment of dorsal and lumber spinal tuberculosis. MethodsFrom March 1996 to July 2000,posterolateral autograft bone graft fusion transpedicular screw system internal fixation and anterior debridement procedures were used in 62 patients suffering from dorsal and lumber spinal tuberculosis in our department,48 of them were involved in a longitudinal study follow-up for a mean of 3.6(1.5-5.5)years postoperatively. ResultsAll patients showed successful posterolateral bone graft fusion. Among 38 cases of Pott's paraplegia, 30 were completely recovered,5 were partly recovered,the rate of recovery was 92.1%. The average immediate postoperative correction of kyphosis angle was 29.1°,the average loss of correction was only 3.2°at final follow-up.ConclusionsPosterolateral autograft bone graft fusion transpedicular screw system internal fixation and anterior debridement procedure were found helpful in strengthening the stability of the spine in dorsal and lumber spinal tuberculosis, providing successful interbody fusion and recovery of Pott's paraplegia, correcting the kyphosis, and preventing progression of kyphosis.

Objective:To explore the effect of estrogen-related receptor α (ERRα) on lipopolysaccharide (LPS)-induced vascular endothelial cell apoptosis and tight junction protein degradation.Methods:RPMVECs transfected with shERRα were cultured in vitro and divided into four groups: Normal control group (Ctr group); shERRα knockdown group (shERRα group); normal cells + LPS treated group (LPS group): The cells in the six-well plates were cultured in serum-free medium for 12 h, and then treated with 20 μg/mL LPS for 12 h; and shERRα+LPS group: ERRα knockdown cells were treated as the LPS group. ROS fluorescence kit was used to detect the intracellular ROS levels . Apoptosis ratio was detected by TUNEL staining, AnnexinV-FITC and PI. Cell membrane ZO-1 expression was detected by cellular immunofluorescence, and the levels of apoptosis-related proteins Bcl-2, Bax, Smac, Cytochrome c, and tight junction protein ZO-1, as well as the expression of Occludin, JAM-A and E-Ca at molecular level were detected by Western blot.Results:Compared with the Ctr group and the shERRα group, the ROS level, apoptosis rate (TUNEL test: 16.44 ± 2.55; and flow cytometry test: 23.56 ± 2.22), the expression of pro-apoptotic proteins Bax, Smac and Cytochrome c were increased in the LPS group, while the expression of anti-apoptotic proteins Bcl-2 and tight junction protein were decreased. In the LPS group. Cellular immunofluorescence results showed that the ZO-1 was degraded in the cell membrane and the network structure was broken. Compared with the LPS group, inhibition of ERRα in the shERRα+LPS group increased cell damage.Conclusions:ERRα can negatively regulate the apoptosis and affect the function of pulmonary microvascular endothelial cells, thereby regulating sepsis-induced acute lung injury.

Objective To evaluate the feasibility and safety of a delivery pathway for the performance of percutaneous left atrial appendage (LAA) occlusion in experimental canine models. Methods Transseptal puncture was performed via femoral vein approach under fluoroscopic and angiographic guidance in 12 experimental dogs. A pigtail catheter was advanced into the left atrium (LA), which was followed by LA angiography. The diameters of the neck of LAA were measured on LAA angiogram obtained in appropriate projection. After the delivery sheath was advanced along the wire into LA, a pigtail catheter was inserted into the ostium of the LAA and the sheath was then advanced over the pigtail into the LAA. LAA angiography was then performed through the delivery sheath to confirm the position of the delivery sheath. One hour after the procedure both electrocardiography (ECG) and transthoracic echocardiography (TTE) were carried out in five dogs to check the results, immediately after which the five dogs were sacrificed to macroscopically observe the damages of the puncture site of inter-atrial septum as well as inside the LA and LAA. One hour and 2 weeks after the procedure TTE was conducted in the remaining 7 dogs and these dogs were followed up for one month. Results One dog died of pericardial tamponade during the operation. In 8 dogs the LAA was clearly displayed in the projection position of right anterior oblique (RAO) 30°/cranial (CRA) 20°,while in 3 dogs the LAA was well visualized in the projection position of RAO 30° , and in one dog in the projection position of RAO 30°/caudal (CAU) 20°. The diameter of LAA neck was (13.6 ± 5.2) mm. The delivery sheath was safely advanced into the LAA along the pigtail catheter in all dogs, and no air embolism, thrombus or pericardial tamponade occurred. Hematoma at puncture point of groin occurred in 2 dogs, which was absorbed through pressure dressing. Macroscopic examination of the heart performed immediately after the operation showed that no bloody pericardial effusion was found, and mild hematoma at posterior wall of LA was seen in one dog and mild damage of the upper-margin intima of LAA was noted in 2 dogs. The mean fluoroscopy time was (10.1 ± 2.5) minutes and the mean operation time was (58 ± 12) minutes. TEE showed no pericardial effusion 2 weeks after the procedure. During the follow-up period of one month no sudden death, stroke or infection occurred. Conclusion This method of placing the delivery sheath into the LAA is clinically safe and effective, and it can reliably establish a pathway to advance the LAA occluder into LAA.

Objective To investigate the diagnostic and clinical significance of computerized tomography (CT) attenuation values (hounsfield unit,HU) in hydronephrosis with infection.Methods One hundred and eighty-five cases of upper urinary tract calculi with hydronephrosis from June 2014 to June 2016 were retrospectively analyzed.There were 82 males and 103 females with a mean age of (52.3 ± 13.1)years old,ranging 18-80 years old.58 cases suffered hydronephrosis without infection,55 cases suffered acute pyelonephritis and 72 cases suffered pyonephrosis.The CT attenuation values of the renal pelvis urine in three groups were measured.Results The CT attenuation value of hydronephrosis without infection group was (5.61 ± 3.67) HU,95 ％ CI(4.64-6.57) H U.In acute pyelonephritis group,CT attenuation value was (8.35 ± 5.63) HU,95％ CI(6.83-9.87) HU.In pyonephrosis group,the CT attenuation value was (13.92 ± 6.21) HU,95％ CI (12.46-5.38) HU.The CT attenuation value of pyelonephritis compared with that of hydronephrosis without infection was significant different.(P ＜ 0.01).The CT attenuation value of the patients with pyonephrosis was significantly higher than that of patients without infection and with pyelonephritis (P ＜ 0.01).Conclusions The CT attenuation value of renal pelvis urine can predict intrarenal infection.Furthermore,The measurement of CT attenuation value has some clinical significance in preoperative evaluation of hydronephrosis with infection.