OBJECTIVE:To draw a comparison among four extraction methods of total DNA from Corydalis saxicola in order to get high quality DNA sample.METHODS:The total DNA was extracted by palmityl trimethyl ammonium bromide method,enzyme-added detergent method,guanidinium isothiocyanate method,inertmacromolecular adsorptive column method,respectively.The purity(defined as RA260/A280)and the yield of the total DNA were determined.RESULTS:The RA260/A280 values of the DNA samples extracted by 4 methods were 1.832 9,1.699 2,1.812 2 and 1.813 6,respectively;and the extraction rates were 140.20 ?g?g-1,69.56 ?g?g-1,132.50 ?g?g-1 and 143.12 ?g?g-1,respectively.CONCLUSION:DNA samples extracted in 4 methods were all of high quality.The yield of DNA sample in enzyme-added extraction method was lower,while the guanidinium isothiocyanate method and the adsorption column method were rapid and simple and they are suitable for the test of samples of small size.

OBJECTIVE: To establish an HPLC method for the determination of Baicalin in Tanreqing injection.METHODS: The determination was performed by HPLC using an CLC-ODS C18 column(150 mm?6.0 mm,5 ?m),with mobile phase consisted of methanol-water(47∶53) at a flow rate of 0.5 mL?min-1 and a detection wavelength of 280 nm.RESULTS: The calibration curve for Baicalin was linear in the concentration range of 2.53～12.65 ?g?mL-1(r=0.999 8).The average recovery was 99.61%(RSD=0.35%,n=9).CONCLUSION: The method is simple,rapid and applicable for the quality control of Tanreqing injection.

OBJECTIVE:To establish HPLC method for the determination the centent of tanshinone Ⅱ_A in danshen shuxin capsule.METHODS:The determination was performed on Shim-pack CLC-ODS column,the mobile phase consisted of methanol-water(15∶5),the detection wavelength of270nm,column temperature at40℃,a flow rate of1.5ml/min and.RE_ SULTS:The calibration curve of tanshinone Ⅱ_A is linear when its concentration range was with in 0.05125～0.25625?g/ml(r=0.9996,n=5),the average recovery was 99.05%(RSD=0.92%).CONCLUSION:This method is convenient in operation,precise,reliable in results,and it can be used as quality control for danshen shuxin capsule.

Objective To investigate the expression of interleukin 6 (IL-6) in tumor microenvironment of natural killer/T cell lymphoma (NKTCL) and its relationship with clinical characteristics and prognosis. Methods From January 2005 to December 2012, 93 patients with NKTCL and available paraffin-embedded tissue in Sun Yat-sen University Cancer Center were included. The expression or IL-6 in tumor microenvironment was investigated by immunohistochemistry. The positive IL-6 expression was identified as≥10 cells/HP. The enumeration data and measurement data were compared by t test andχ2 test, respectively. Overall survival (OS) and progression free survival (PFS) were estimated with the Kaplan-Meier method. Survival rates were compared by the Log-rank test. Statistical significance was determined at a level of P<0.05. Results The median count of IL-6 positive cell number was 21 cells/HPF (range, 0-150 cells/HP), and 61.29%(57/93) patients were positive. Positive IL-6 expression was mostly associated with fever and high Korean prognostic index (KPI) score (both P< 0.05). The serum c-reactive protein (CRP) level was (29.28 ± 2.62) mg/L in IL-6 positive patients and (11.14±2.77) mg/L in IL-6 negative patients (t= -2.276, P= 0.025). Patients with negative IL-6 expression had better survival, with 52.7 % of 5-year PFS rate,and 60.0 % of 5-year OS rate; the 5-year PFS and OS rates in those with IL-6 positive were 23.6 % and 27.1 %, respectively (both P= 0.001). Conclusion The high expression of IL-6 in patients with NKTCL might be associated with adverse clinical feature and poor survival.

Ginko biloba extract is an active compound with broad biological function.This review summarizes the immuomodulatory effect of Ginko biloba extract on immune organ development, monocyte macrophage, natural killer cell, humoral immunity, cellular immunity, cytokines, red cell immunity and mucosal immunity.

Objective To investigate the effects of exercise on knee cartilage tissue structure using quantitative MR T2 mapping. Methods Sagittal T2 maps of the knee joints of 26 healthy volunteers were obtained by using 3.0T MR before,immediately after, and 1 5 min after running.The original images were classified into three terms of knee cartilage T2 map after postreconstruction.The T2 values of regions of interest (ROC)(T2 pre ,T2 post ,T2 delay )in the superficial,middle and deep cartilage of femoral and tibial joint were measured.Statistical differences of cartilage T2 values of three terms after running were analyzed.Results For the tibial joint cartilage,the T2 pre ,T2 post ,T2 delay were (49.71 ± 1.95)ms,(44.30 ± 2.56)ms,(49.41 ± 1.62)ms in the superficial layer,respectively.The three terms T2 were (42.43 ± 2.23)ms,(39.01 ± 2.37)ms,(41.90±2.28)ms in the middle layer,respectively.The differences were statistically significant(F=55.673,16.759 respectively.P<0.001).While the three terms T2 were (19.39±2.13)ms,(19.20±2.22)ms, (19.49±2.05)ms in the deep layers cartilage,respectively.The differences were not statistically significant(F =0.122,P =0.886).And the differences between T2 pre and T2 post ,T2 post and T2 delay were statistically significant (all P <0.001)in superficial and middle alyers,but there were no significant difference between the T2 pre and T2 delay (P =0.610,0.403,respectively).For the femoral joint cartilage,the T2 pre ,T2 post ,T2 delay were (50.22 ± 1.47)ms,(45.60 ± 2.82)ms,(49.84 ± 1.84)ms in superficial layers,respectively.The three terms T2 were (42.67±2.23)ms,(39.36 ± 1.98)ms,(42.40 ± 2.57)ms in the middle layer,respectively.The differences were statistically significant (F=37.976,16.987 respectively,P<0.001).While the three terms T2 were (20.30±2.73)ms,(20.60±2.44)ms,(20.51± 2.24)ms in the deep layer,the differences were not statistically significant (F =0.098,P =0.907).And the differences between T2 pre and T2 post ,T2 post and T2 delay were statistically significant (all P <0.001)in superficial and middle layers,but there were no significant difference between the T2 pre and T2 delay (P=0.520,0.679,respectively). Spatial distribution of T2 values of articular cartilage from deep to superficial layers showed a ascending trend.T2 maps showed the spatial distribution trend of T2 value change.Conclusion T2 mapping can monitor quantitatively the changes of articular cartilage molecular structure after running.The change of articular cartilage T2 value after exercise is uneven and the change of articular cartilage structure after exercise is reversible.

Objective To investigate the clinical feasibility of contrast?enhanced three dimensional T2WI turbo?spin?echo sequence with short?term inversion recovery and sampling perfection using different flip angle evolutions (3D STIR T2WI SPACE) sequence in the brachial plexus neurography. Methods Thirty two patients were prospectively chosen and performed with brachial plexus plain scanning on a 3.0 T MR scanner by using plain and contrast?enhanced 3D STIR T2WI SPACE sequence. Thirteen of them underwent plain scan, 9 of them underwent contrast?enhanced scan, and 10 of them underwent both plain scan and enhanced scan. The visibility of the brachial plexus were scored and contrast to noise ratio (CNR) were measured by two experienced radiologists. The results between plain and contrast?enhanced imaging were compared by t test. The 10 subjects received both enhance and plain imaging, were performed with paired t test. Results In 32 patients, the visibility score of brachial plexus nerve and CNR were 7.8 ± 1.3 and 24.97±3.41 in the plain scan group, and 13.1±1.7 and 38.49±4.95 in enhanced scan group, respectively. There were statistically significant differences in the two groups(t=-11.72,P<0.01;t=-10.47, P<0.01). In 10 cases with plain and enhanced brachial plexus imaging, the average score of the brachial plexus were 7.4 ± 1.7 and 13.3 ± 1.6, the average CNR were 26.23 ± 4.43 and 38.19 ± 5.03 respectively. There were statistically significant differences (t=- 8.22, P<0.01; t=- 5.64,P<0.01). The score results were analyzed for consistency. Plain images Kappa value was 0.684, which shows moderate consistency and enhanced images Kappa value= 0.822, which shows excelent consistency. Conclusions The contrast?enhanced 3D STIR T2WI SPACE sequences may suppress background tissue signals, which is helpful to display brachial plexus, therefore it is of important value for the early diagnosis of brachial plexus neuropathy.

OBJECTIVETo establish a method of methyl-DNA immunoprecipitation (MeDIP)-real time quantitative PCR (qPCR) for detecting the promoter methylation level in cell-free seminal DNA (cfsDNA).

METHODSWe obtained cfsDNA samples from 6 normozoospermia men (the NZ group) and 6 post-vasectomy patients (the PV group), and mixed the samples from different individuals of each group, respectively. Then we made DNA fragments by ultrasonication, separated the methylated DNA fragments by MeDIP, and determined the methylation level of the promoters in cfsDNA by qPCR.

RESULTSThe methylation levels of the promoters PRAME, PEG10, MORC1, GML, HOXA5, DNMT3L, SNURF, MSH4, DAZ1 and CLPB were 14.93, 2.64, 0.69, 2.66, 17.50, 21.10, 5.98, 2.28, 13.50 and 3.86%, respectively, in the NZ group, obviously lower than 121.25, 73.62, 16.25, 42.90, 76.74, 112.40, 59.79, 25.85, 91.90 and 64.53% in the PV group. The results of MeDIP-qPCR for the methylation of PRAME, MORC1, GML, HOXA5, DNMT3L, SNURF, MSH4 and DAZ1 were coincident with the results of genome-wide promoter methylation microarray.

CONCLUSIONMeDIP-qPCR can quantitatively measure the promoter methylation level in cfsDNA, and effectively determine the testis- and epididymis-specific methylated promoters in human semen.

Adult ; DNA ; chemistry ; DNA Methylation ; Epididymis ; metabolism ; Epigenesis, Genetic ; Humans ; Male ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Semen ; chemistry ; Testis ; metabolism