Objective To establish and evaluate a universal real-time fluorescent quantitative PCR(qPCR)method for identifying and quantifying three human parvovirus B 19 ( B19V) genotypes.Methods Firstly, following a bioinformatic analysis of a subset of B19V genomic sequences available in the NCBI nucleotide database ,representative of genotypes 1 to 3,a set of suitable universal primers and TaqMan probes was designed from the NS 1 gene of B19V.Aplasmid was used as a quantitative standard that contained the identical sequence of the B 19 target sequence .An internal control ( IC ) was included to prevent false negative results .Then,serial 1-log dilutions of quantitative standards were prepared and used in the qPCR assays for generation of a standard curve .Finally,the specificity,sensitivity and reproducibility of the assay were assessed.Results A linear relationship of the real-time PCR method for detecting B19V from 1 ×109copies/μl to 1 ×103 copies/μl was observed .The developed qPCR protocols allowed for the detection of genotypes 1 to 3 with a limit of detection ( LOD) of 10 copies/μl.Furthermore, the assay did not amplify other blood-borne viruses.The inter-and intra-assay variability analyses showed good reproducibility of the assay .Conclusion A universal real-time qPCR method for the detection of B19V DNA is established,which will facilitate the diagnosis of B19V infections and the screening of blood and plasma-derived products , thereby improving the viral safety of transfusion and plasma-derived products .

AIM: A HPLC fingerprint has been established of six active principles in Cistanche tubulosa in nine different regions in the Xinjian Uygur Automomous Region in China. METHODS: Chromatographic condition included a symmetry C_(18)(4.6 mm?250 mm,5 ?m) column and the gradient elution was adopted with ratio of acetonitrile-0.095% H_3PO_4 from 8()∶92 to 12()∶88 in 0-18 min and 12()∶18 to 19()∶81 in 18-40 min,19()∶81 maintain for 50 min.The detection wavelength was at 330 nm and the flow rate was 1.0 mL/min and detection time was 90 min. RESULTS: Eleven common characteristic peaks,including salidroside, bartsioside,echinacoside,cistanbuloside A,tabuloside A and acteoside were taken as fingerprint peaks the precision and the accuracy were in accordance with the chromatographic requirement. CONCLUSION: The method is stable,reliable,precision and provides a scientific basis for the quality standard for Cistanche tubulosa.

Objective To detect human parvovirus B19(B19V)DNA in source plasma pools and coagulation factor products and determine its prevalence and the level of contamination .Methods A pair of primers and a probe selected from the highly conserved sequences encoding the non-structural protein(NS1)of B19 were designed and synthesized.With the primer-probe combination ,source plasma pools and four types of coagulation factor products were determined for B 19V DNA by TaqMan real-time quantitative PCR.Results One-hundred and sixteen from 195 (59.49%) source plasma pools contained B19 DNA and concentrations up to 1.35 ×1010 copies/ml were measured.High frequencies of contamination were detected in factor Ⅷ (29 of 31; 93.55%), thrombin (10 of 10; 100%), fibrinogen (6 of 7; 85.71%) and prothrombin complex (8 of 9;88.89%).Conclusion These data show that B19V is a common contaminator in Chinese source plasma pools and coagulation factor products .Thus,B19V screening in Chinese source plasma seems desirable and significant for the safety of plasma derivatives in China .

The genetic diversity of three Tibetan herbs, i. e., Sang-Di, E-Dewa and Ye-Xingba (Tibetan names), was studied based on the field collection, specimen identification and DNA sequence analysis. Swertia hispidicalyx, Gentiana lhassica and Scrophularia dentata, as the original plants of the three Tibetan herbs, were collected and identified. The regions of ITS, matK, rbcL, rpoC1, trnL(UAA), psbA-trnH, atpB-rbcL, trnS (GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF(GAA) and nadl 2nd intron were amplified and sequenced. The ITS regions of S. hispidicalyx and S. dentata were cloned and sequenced, and the sequences were classified into different genotypes. All the sequences were analyzed and compared with those of closely related species. Our studies may provide reference for the genetic diversity analysis and molecular identification of the three Tibetan herbs.
Genetic Variation ; Gentiana ; classification ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Plants, Medicinal ; classification ; genetics ; Scrophularia ; classification ; genetics ; Swertia ; classification ; genetics ; Tibet

Objective To evaluate the imaging performance and characteristics of peripheral primitive neuroectodermal tumors (pPNET), in order to raise awareness of the disease and the diagnostic accuracy. Methods Thirteen patients with pPNET who confirmed by puncture or pathology were enrolled in this study. The CT and MRI features were retrospectively analyzed. Four patients received simple routine CT examination, and 9 patients had MRI examination based on X-ray, CT examination. Results The part of body involved was more widely.Two patients occurred in the sinuses, 1 patient in the chest wall, 1 patient in the lung, and 3 patients in the abdominal and pelvic(1 patient involved the pelvic floor through the pelvic wall lesions to basins), 1 patient in the spine, and 5 patients located in the extremities. The characteristics of pPNET by CT and MRI in soft tissue of 8 patients showed single large soft tissue mass, and the biggest was 11.2 cm × 10.6 cm. In bonds and joints of 5 patients, CT and MRI showed wide range of soft tissue mass, mixed internal signal/density with obviously uneven enhancement, significant bone destruction and dissolvable osseous changes, and no obvious tumor bone and calcification. Conclusions Each site has its relative imaging characteristics of pPNET. CT and MRI can show lesions involving the scope, internal structure, with or without adjacent tissue invasion and metastasis, and provide guidance for clinical treatment and selecting treatment regimen. But in the end, it depends on the diagnosis of pathological and immunohistochemical examination.

[ ABSTRACT] AIM: To investigate the seroprevalence of neutralizing antibodies to human adenovirus type 5 (AdHu5) , human adenovirus type 26 (AdHu26) and chimpanzee adenovirus type 68 (AdC68) in the patients with chro-nic hepatitis B ( CHB) and the patients with primary liver cancer ( PLC) , and to provide guidance for developing safe and effective biotherapy vectors against CHB and PLC.METHODS:The blood samples from 196 patients with CHB and 193 patients with PLC were examined to assess the presence of neutralizing antibodies against AdHu5, AdHu26 and AdC68 by adenovirus neutralization assays.RESULTS:The seroprevalence rates of neutralizing antibodies to AdHu5, AdHu26 and AdC68 in the CHB patients were 84.7%, 58.2%and 39.8%, respectively.Among the patients with PLC, the prevalence rates of neutralizing antibodies were as follows:AdHu5, 75.1%;AdHu26, 66.8%;AdC68, 32.1%.CONCLUSION:The prevalence rates and titers of neutralizing antibodies against AdC68 were the lowest among the 3 adenoviruses.There-fore, AdC68 serves as more suitable biological therapy vectors for CHB and PLC than AdHu5 and AdHu26.

In order to clarify the chemical constituents in Qiliqiangxin capsule, a rapid ultra-performance liquid chromatography/orthogonal acceleration time-of-flight mass spectrometry (UPLC-Q-TOF/MS(E)) method was established. Forty peaks were identified on line using this method. The herbal sources of these peaks were assigned. The results implied that triterpenoid saponins, flavonoid glycosides, C21-steroids and phenolic acids were included in the main components of Qiliqiangxin capsule. The method is simple and rapid for elucidation of the constituents of Qiliqiangxin capsule and the results are useful for the quality control of Qiliqiangxin capsule.

This study aims to construct a dynamic two-dimensional characterization technique for the hygroscopicity of traditional Chinese medicine extracts and investigate the effect of material properties of powders on hygroscopicity. The dynamic hygroscopicity-time curves of the powders were measured at 25 ℃ and 75% humidity, and the semi-equilibrium hygroscopicity time (t_1/2) and equilibrium hygroscopicity (F^∞) were derived as two-dimensional evaluation indicators. Finally, the correlation between the material properties and the hygroscopic behavior was analyzed by principal component analysis (PCA) and partial least squares analysis (PLS). The results showed that the dynamic two-dimensional characterization system of hygroscopicity constructed with 1/t_1/2 = 0.1 h^-1 and F^∞ = 15% as the center can classify the hygroscopic behavior of traditional Chinese medicine extracts into four categories: fast hygroscopicity with strong hygroscopicity, slow hygroscopicity with strong hygroscopicity, fast hygroscopicity with weak hygroscopicity and slow hygroscopicity with weak hygroscopicity. The moisture absorption was negatively correlated with D₅₀, D₉₀, ρ_b and ρ_t; the moisture absorption rate was negatively correlated with D₁₀, D₅₀, D₉₀, ρ_b, ρ_t, and positively correlated with moisture content. The hygroscopicity dynamic two-dimensional characterization indicators of Chinese medicine extracts (CMEs) constructed in this study matched with the physical properties. The method of dynamic multi-dimensional characterization technology is feasible and scientific, and the idea has strong promotional value.

OBJECTIVETo investigate the effects of progesterone on the growth and migration of breast cancer cells.

METHODSMCF-7 and T-47D cells were cultured in DMEM and stimulated with 100 nmol/L progesterone for 48 h, and the cell proliferation was evaluated by MTT assay, cell migration by wound-healing assay and E-catherin expression by Western blotting.

RESULTSProgesterone stimulated the cell proliferation and migration and down-regulated the expression of E-catherin in both MCF-7 and T-47D cells.

CONCLUSIONSProgesterone stimulates the cell proliferation and migration of cultured breast cancer cells, suggesting the clinical significance of anti-progesterone therapy in breast cancer.

Breast Neoplasms ; pathology ; Cadherins ; metabolism ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Female ; Humans ; Progesterone ; pharmacology ; Tumor Cells, Cultured

OBJECTIVETo evaluate a technique for closing the cleft of nasal aspect at the junction of the hard and soft palate and ensuring the wound healing well during the procedures of repairing broad cleft palate.

METHODSA inferior turbinate mucoperiosteal flap based posterior is designed and transposed to the junction of the hard and soft palate for decreasing the tension effectively and closing the cleft easily.

RESULTSAll of the patients showed wound healing well and no distinct scars or palatal fistula.

CONCLUSIONSIt is a simple and effective technique to decrease the tension and close the cleft of nasal aspect at the junction of the hard and soft palate with inferior turbinate mucoperiosteal flap.

Child, Preschool ; Cleft Palate ; surgery ; Humans ; Male ; Periosteum ; transplantation ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Turbinates