OBJECTIVE：To establish the method for content determination of 6 active ingredients in Paeonia lactiflora during different harvest periods ，and to investigate its variation rules so as to determine the optimal harvesting period. METHODS ：HPLC method was adopted to determine the contents of gallic acid ， catechin， alibiflorin， paeoniflorin， benzoic acid and benzoylpaeoniflorin，and principal component analysis was conducted . The determination was performed on Thermo C 18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid aqueous solution （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was 230 nm，and column temperature was 35 ℃. The sample size was 10 µL. RESULTS ：The linear range of the above 6 ingredients were 0.013 2-0.25 mg/mL（r＝0.999 2），0.013 2-0.25 mg/mL（r＝0.999 9），0.026 8-0.51 mg/mL（r＝0.999 7）， 0.42-8.01 mg/mL（r＝0.999 2），0.016-0.31 mg/mL（r＝0.999 4），0.02-0.38 mg/mL（r＝0.999 8），respectively. The limits of quantification were 0.009 3，0.008 5，0.016 3，0.021 7，0.011 3，0.017 4 mg/mL，and the limits of detection were 0.003 3，0.002 7， 0.005 4，0.007 3，0.003 8，0.005 9 mg/mL. RSDs of precision ，stability and repeatability tests were less than 2％. The recoveries were 96.01％-99.43％（RSD＝1.23％，n＝9），97.95％-100.45％（RSD＝0.79％，n＝9），97.98％-100.11％（RSD＝0.68％，n＝ 9），98.83％ -100.09％ （RSD＝0.65％ ，n＝9），98.58％ - 100.95％（RSD＝1.35％，n＝9），96.28％-103.26％（RSD＝ 1.76％，n＝9）. The contents of above 6 ingredients in Radix Paeoniae Rubra （roots） were 0.016％ -0.057％ ，0-0.067％ ， 0.207％-0.640％，2.350％-5.887％，0.030％- 0.245％，0.054％- 0.381％，respectively. On May 30th，the drying rate of Radix 0451-87266873。E-mail：wangzhen_yue@163.com Paeoniae Rubra was the lowest （about 33％），and on Sept. 15th，the drying rate was the highest （about 49％）. The contents of gallic acid and paeoniflorin in the leaves of P. lactiflora were higher than the root during Jul.-Oct. Results of principal component analysis showed that the variance contribution rates of the first two principal components were 71.845％ and 18.170％，respectively；cumulative variance contribution rate was 90.015％. The months with higher comprehensive scores were May to Jun. and Sept. to Oct. CONCLUSIONS ：Established method is simple ， accurate，reproducible and precise. It can be used to determine the contents of 6 active ingredients in Radix Paeoniae Rubra during different harvest periods. Sept. 30th to Oct. 15th is the optimum harvesting periods for Radix Paeoniae Rubra ，and leaves can be harvested around Jul. 15th.

Objective: To investigate the prognostic value of N-terminal B-type natriuretic peptide (NT-proBNP) on all-cause mortality in heart failure patients with preserved ejection fraction (HFpEF) at real world scenarios. Methods: Patients who met the diagnostic criteria of HFpEF in the China National Heart Failure Registration Study (CN-HF) were divided into death and survival groups. The demographic data, physical examination, results of the first echocardiography, laboratory results at admission, complications, drug use and clinical outcomes were obtained from CN-HF. The univariate Cox proportional hazard model was used to screen the variates that might predict prognosis, and then the covariates with statistical significance were included in the multivariate Cox regression model to analyze the predictive value of baseline NT-proBNP on all-cause death. Spearman correlation analysis was used to evaluate the relationship between NT-proBNP and estimated glomerular filtration rate (eGFR), so as to further explore the predictive value of the interaction between renal dysfunction and NT-proBNP on death. Since NT-proBNP did not obey the binary normal distribution, it was expressed by the natural logarithm of NT-proBNP (LnNT-proBNP). Results: A total of 1 846 HFpEF patients were enrolled in this study, with an average age of 71.5 years, 1 017 males(55.1%), median NT-proBNP 860 ng/L, and median eGFR 73.9 ml·min^-1·1.73m^-2. After a median follow-up of 34 months, 213 (11.5%) patients died. Patients in the death group were older, with higher NYHA classification Ⅲ-Ⅳ ratio, longer hospital stay, higher serum potassium and NT-proBNP level, prevalence of complications of diabetes mellitus, arrhythmia and atrial fibrillation, use of angiotensin receptor antagonist(ARB), mineralocorticoid receptor antagonists (MRA), diuretic and digoxin was significantly higher in death group than in survival group. Body mass index (BMI), diastolic blood pressure, left ventricular ejection fraction (LVEF), hemoglobin, serum cholesterol(TC), serum triglycerides (TG) and eGFR, and use of angiotensin converting enzyme inhibitors (ACEI), statins and aspirin were lower in death group than in survival group. Univariate Cox regression analysis showed that NT-proBNP was a predictor of all-cause death in HFpEF patients (HR=2.522, 95%CI 2.040-3.119, P<0.001). Multivariate Cox regression analysis showed that the elevated NT-proBNP remains as the independent predictor of all-cause death in patients with HFpEF (HR=1.230, 95%CI 1.049-1.442, P=0.011) after adjusting for age, BMI, diastolic blood pressure, LVEF, hemoglobin, serum potassium, serum sodium, TC, serum high-density lipoprotein cholesterol (HDL-C), TG, eGFR, atrial fibrillation, as well as the treatment of ACEI/ARB, MRA, diuretics and digoxin. Spearman correlation analysis showed that LnNT-proBNP was negatively correlated with eGFR (r=-0.361, P<0.001), but there was no interaction between NT-proBNP and renal dysfunction in predicting death in HFpEF patients (P>0.05). Conclusion The elevated level of NT-proBNP at admission is an independent predictor of all-cause mortality in HFpEF patients.

OBJECTIVE： To establish a determination method for content of polysaccharide in Acanthopanax senticosus， and to conduct cluster analysis and ultrasonic extraction technology optimization. METHODS： The content of polysaccharide in A. senticosus was determined by phenol-sulfuric acid method. Cluster analysis was conducted by using SPSS 23.0 software of the polysaccharide in A. senticosus from 17 habitats. The ultrasonic extraction technology was optimized with L9（34）orthogonal test using extraction temperature， the ratio of material to liquid， extraction time as factors， the content of polysaccharides as index， and then validated. RESULTS： The linear range of glucose ranged from 0.007 75 to 0.151 mg/mL （r＝0.999 1）； the limits of quantification and detection were 2.854， 0.856 μg/mL； RSDs of precision， stability， and repeatability tests were less than 2％； recovery rates of the sample were 98.41％-101.58％（RSD＝1.23％，n＝6）. Cluster analysis results showed that 17 batches of samples could be clustered into three classes； S1， S6， S10， S12 and S13 were clustered into one class； S3-S5 and S7 were clustered into one class； and the rest samples were clustered into one class. The optimal ultrasonic extraction technology was extraction temperature 55 ℃， ratio of material to liquid 1 ∶ 10 （g/mL）， extraction time 35 min. Results of validation tests showed the content of the polysaccharide under the best process condition was 4.36％（RSD＝0.920％，n＝3）. CONCLUSIONS： Established method is simple，reproducible and suitable for determination of polysaccharide in A. senticosus； the optimized ultrasonic extraction technology is stable and feasible.

Objective To investigate the effects of spine region of interest (ROI),tracking range,and real-time image contrast ratio on the positioning errors of cyberknife.Methods The LTT dynamic phantom was used to develop a spine tracking plan and perform treatment,and the target positioning system was used to preset the phantom and obtain real-time preset image and positioning error.Based on the realtime preset image,spine ROI,tracking range,and real-time image contrast ratio were changed to observe the changes in positioning error and related parameters.Pearson correlation analysis was performed.Results The change in tracking range did not change the positioning error in spine tracking,and tracking range was not correlated with positioning error (R =0,P =1).The changes in ROI and image contrast ratio did not affect the translation error,but affected the rotation error,especially the rotation error in left-right direction (r =0.533 and 0.693,P=0.002 and 0.026).The image contrast ratio had the most obvious effect,with an amplitude of variation up to 2.2°.Conclusions The change in tracking range does not affect the positioning errors in spinal tracking,but the changes in ROI and image contrast ratio can cause varying degrees of changes in positioning errors,which should be taken seriously in cyberknife treatment.

Objective To analyze the thin-section CT features of fissure-attached nodules in asymptomatic patients.Methods The baseline and follow-up CT images of 120 asymptomatic patients with fissure-attached nodules were retrospectively analyzed.The thin-section CT findings include size,shape,density,interface,and the relationship with pulmonary fissure.Results There were 1 67 fissure-attached nodules of the 120 patients.The mean length of nodules was 5.46 mm±2.276 mm (2 - 1 5 mm).104(104/1 67,61.1%)nodules were less than 5 mm in length.1 63 (1 63/1 67,97.6%)nodules were typical and 4 (4/1 67,2.4%)nodules were atypical.All these nodules manifested as homogenous,solid,smooth interface,and widely adjacent to the fissure on CT ima-ges.Among those nodules,1 64(1 64/1 67,98.2%)nodules were triangular(88/1 67,52.7%)or oval(76/1 67,45.5%).126(126/1 67, 75.4%)nodules were abutting the major fissure.The angle between nodule and fissure was obtuse in 1 64(1 64/1 67,98.2%)fissure-attached nodules.During follow-up,4(4/1 67,2.4%)nodules increased in size and the corresponding volume doubling time (VDT) range was 863-4 228 days.However,no obvious malignant signs were found.Conclusion The thin-section CT images of fissure-attached nodules in asymptomatic patients showed the benign characteristics,most of which less than 5 mm and demonstrated trian-gle or oval,smooth interface and homogenous solid density.

Objective MiR-375 is lowly expressed in esophageal squamous cancer cells and the downstream target gene of miR-375 remains unclear .This paper discusses the role of miR-375 in regulating the expression of short stature homobox 2 ( SHOX2) in human esophageal squamous cancer cells . Methods The bioinformatics software TargetScan , miRanda, PicTar, miRTarget2, and PITA were used to predict the assumptive targets of miR-375 in SHOX2.Then, two recombinant luciferase gene report plasmids containing wild pSHOX2 3′UTR ( pSHOX2-375-WT ) and mutant pSHOX2 3′UTR ( pSHOX2-375-mut ) were constructed , sequenced , and identified.Human esophageal squamous cancer cells were co-transfected with miR-375 mimics and pSHOX2-375-WT or pSHOX2-375-mut, respectively , and divided into 7 groups: pmiR, pSHOX2-375-WT, pSHOX2-375-WT +miR-375, pSHOX2-375-WT +miR-NC, pSHOX2-375-mut, pSHOX2-375-mut+miR-375, and pSHOX2-375-mut+miR-NC, each subjected to the measurement of luciferase activity .The expressions of SHOX 2 mRNA and protein were de-termined after transfection of the esophageal squamous cancer cells with miR-375 mimics, and so were the expressions of miR-375 and SHOX2 in the esophageal squamous carcinoma tissue samples obtained postoperatively . Results Prediction with the five software showed only one conserved function site of miR-375 in SHOX2 3′UTR at 1156-1170 bp.Luciferase activity was significantly lower in the pSHOX2-375-WT+miR-375 group (0.261 ±0.036) than in the pmiR (1.818 ±0.061), pSHOX2-375-WT (1.820 ±0.086), pSHOX2-375-WT+miR-NC (1.851 ±0.094), pSHOX2-375-mut (1.861 ±0.059), pSHOX2-375-mut +miR-375 (1.896 ± 0.048), and pSHOX2-375-mut+miR-NC group (1.760 ±0.062) ( P<0.01).SHOX2 mRNA and protein expressions were sup-pressed by the overexpression of miR-375 in the EC9706 cells.The expression of miR-375 was decreased, while that of SHOX2 in-creased in the esophageal squamous carcinoma tissue as compared with the normal esophageal tissue . Conclusion MiR-375 regu-lates the expression of the SHOX 2 gen in esophageal squamous cancer cells .

Objective To explore the relationship of X-ray exposure parameters and false-node rate during image-guidance treatment with CyberKnife spine tracking.Methods Using spine tracking planning on a chest phantom,several combinations of X-ray exposure parameters were used to locate.The false-node ratio and the surface absorbed dose were investigated and the radiation dose was optimized.Results The false-node ratio and surface absorbed radiation dose decreased when the X-ray exposure parameters increased until they saturated.In the range of ≤5.0％ false-node rate,the surface absorbed radiation dose was 0.11,0.26 mGy,and 0.31-0.46 mGy,when the false-node rate was 2.77％,1.07％and 1.0％,respectively.Conclusions In image-guided treatment of CyberKnife,the radiation dose would be optimized,and the patient's radiation dose would be reduced greatly,which is important to protect the patients.

Objective To compare the quality and preservation effect among the three models of pooled buffy coats (PBC) in pre-paring the platelet concentrate (PC) .Methods 63 identical ABO blood donations were randomly and averagely divided into three groups :the immediate PBC group(n=21) ,the buffy coats(BC) group(n=21) and the whole blood(WB) group(n=21) .In the im-mediate PBC group ,both the separation of the BC and the preparation of the PC was finished in the first day after collecting the WB;in the BC group ,the separation of BC was executed in the first day after collecting the WB ,while the preparation of the PC was completed in the second day ;in the WB group ,both the separation of the BC and the preparation of the PC was implement in the second day after collecting the WB .All the prepared PC were storage at (22 ± 2) ℃ for seven days in the preservation solution (composed by 2/3 PAS-ⅢM and 1/3 plasma) .Then compare the platelet(PLT) counts ,the red blood cell(RBC) residual quanti-ties ,the aggregative function of PLT ,the positive expression rates of CD62p ,the capability of hypotonic shock response(HSR) ,the PH value and the bacterial growth among PCs prepared by the three models during the stored time .Results Compared the immedi-ate PBC group during the seven stored days ,the PLT counts of the BC and WB groups were more ,the aggregative function of PLT in the BC group was better and the capability of HSR in the WB group was stronger ,but the other indexs between the immediate PBC group and the BC or WB group were no significant difference(P>0 .05) .Conclusion Both an overnight holding of BC and WB at room temperature models of PBC to prepare the PC are safe ,reliable and convenient ,and they could be substitute for the immedi-ate PBC model to prepare the PC .

Objective To measure and analyze Varian Clinac 21EX accelerator induced radioactivity,and to provide specific recommendations and ways of radiation protection for radiotherapy technicians.Methods To simulate the working environment of radiotherapy technician,and to detect induced radioactivity variation of Varian Clinac 21EX accelerator that induced by 15 MV X-rays under different conditions of beam field area,dose,time,distance and by high energy electron beam (12,16,20 MeV)at the different time.Results The induced radioactivity level was not influenced by different beam field area,and was increased with increasing dose (r =0.930,P ＜ 0.05),decreased with time increase (r =-0.84,P ＜ 0.05),decreased with distance increase(r =-0.975,P ＜ 0.05).The induced radioactivity attenuation levels of the different doses at the same time are different,and have the common characteristic that the induced radioactivity attenuation rate is faster in the initial times.The induced radioactivity levels of high energy electron beams were significantly lower than those of high-energy X-rays.Conclusions During radiotherapy positioning,it is necessary to take measures to protect against the induced radioactivity when high-energy rays with energy greater than 10 MeV will be used.The radiotherapy technician should take the different time and energy interval steps for the different ray type and energy and dose in order to meet the radiation protection principle of optimization.

OBJECTIVE: To analyze the clinical and genetic features of a patient with Treacher Collins syndrome (TCS), and identify the mutation in TCOF1 gene. METHOD: The medical history was taken, and general physical examinations and otological examinations were conducted in this patient. Genomic DNA was extracted from this patient and his parents and complete TCOF1 gene coding exons were amplified by specific PCR primers. Direct sequencing was carried out to identify the mutations. The raw data was analyzed with GeneTool software and molecular biological website. RESULT: We detected a heterozygous c. 1639 delAG mutation in exon 11 of TCOF1, which resulted in a truncated protein lacking normal function. This mutation is a novel mutation and the second case identified in exon 11 of in TCS. CONCLUSION TCS patient reported in this study has unique clinical phenotype. TCOF1 gene mutation is the specific risk factor.
DNA Primers ; Exons ; Genes, Regulator ; Genetic Testing ; Humans ; Mandibulofacial Dysostosis ; diagnosis ; genetics ; Mutation ; Nuclear Proteins ; genetics ; Phenotype ; Phosphoproteins ; genetics ; Syndrome