The total triterpene saponins of Psammosilene tunicoides have significant pharmacologic activity. Psammosilene tunicoides squalene synthase (PSS) is a gateway enzyme to regulate the biosynthesis of total triterpene saponins extracted from the root of Psammosilene tunicoides which is an endangered species. In this paper, cDNA encoding of PSS was cloned by the degenerate primer PCR and rapid-amplification of cDNA ends (RACE). The full-length of cDNA of PSS is 1663 bp, with an open reading frame (ORF) of 1 245 bp, encoding 414 amino acid polypeptide (calculated molecular mass, 47.69 kDa), 5'UTR (untranslated region) and 3'UTR are 260 bp and 158 bp, respectively. The deduced amino acid sequence of PSS has higher homology with the known squalene synthases of several species such as Panax notoginseng (83%), Panax ginseng (82%) and Glycyrrhiza glabra (82%) than that with Schizosacharomyces pombe (35%), Candida albicans (39%) and Homo sapiens (47%). The characterization of PSS was done by a series of methods, such as prokaryotic expression, the activity of enzyme in vitro, capillary gas chromatography (GC) and capillary gas chromatography mass spectrometry (GC-MS). The results showed that the cell-free extract of E. coli transformed with the recombinant plasmid can effectively convert farnesyl diphosphate into squalene in vitro. GenBank accession number is EF585250. Our research provided important base for the study of Psammosilene tunicoides secondary metabolism and metabolic engineering.
Caryophyllaceae ; enzymology ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Endangered Species ; Escherichia coli ; genetics ; metabolism ; Farnesyl-Diphosphate Farnesyltransferase ; genetics ; metabolism ; Gas Chromatography-Mass Spectrometry ; Open Reading Frames ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; metabolism ; Sequence Homology, Amino Acid ; Transformation, Genetic

To improve the after-sales service, a survey aimed at the after-serveis of 3 kinds of medical equipment is applied among 68 hospitals in Shanghai Area in 2008.The Stat. and analysis results are showed in the paper, which will certainly channel off suppliers to set up a harmonious market together.
China ; Consumer Behavior ; statistics & numerical data ; Data Collection ; Product Surveillance, Postmarketing

OBJECTIVETo evaluate the effect of alprazolam use on psychological status and hospitalization cost in patient with paroxysmal supraventricular tachycardia underwent electrophysiology studies or radiofrequency catheter ablation.

METHODSIn this prospective, randomized, double-blind, placebo-controlled study, 142 inpatients [77 males, mean age (43.1 +/- 14.5) years] were randomly assigned to receive alprazolam (0.4 mg qd at 10PM for 3 days, n = 72) or placebo (n = 70) 3 days before scheduled electrophysiology studies or radiofrequency catheter ablation. All patients were examined by the Chinese version of Symptom Checklist-90 (SCL-90) at 24 hours before the procedure.

RESULTSCompared with the placebo group, the scores of somatization (1.38 +/- 0.40 vs. 1.65 +/- 0.56, P < 0.01), anxiety (1.50 +/- 0.39 vs. 1.69 +/- 0.50, P < 0.05), phobic anxiety (1.24 +/- 0.36 vs. 1.47 +/- 0.57, P < 0.01), psychotism constructs (1.24 +/- 0.34 vs. 1.35 +/- 0.30, P < 0.05) and global severity index (1.36 +/- 0.35 vs. 1.49 +/- 0.37, P < 0.05) were significantly decreased in alprazolam group. The hospitalization costs were also significantly lower in alprazolam group (32 498 +/- 1170) yuan compared to placebo group (32 947 +/- 1096) yuan, P < 0.05.

CONCLUSIONThe alprazolam use before electrophysiology studies and radiofrequency catheter ablation can improve the patients' psychological status and reduce the hospitalization costs.

Adolescent ; Adult ; Aged ; Alprazolam ; therapeutic use ; Anti-Anxiety Agents ; therapeutic use ; Catheter Ablation ; economics ; psychology ; Double-Blind Method ; Female ; Hospitalization ; economics ; Humans ; Male ; Middle Aged ; Prospective Studies ; Tachycardia, Paroxysmal ; psychology ; therapy ; Tachycardia, Supraventricular ; psychology ; therapy ; Young Adult

Adenoviridae ; genetics ; Animals ; Carcinoma, Hepatocellular ; therapy ; Cell Line ; Endostatins ; genetics ; metabolism ; Genetic Therapy ; methods ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Liver ; cytology ; metabolism ; Liver Neoplasms ; therapy ; Plasmids ; genetics ; Polymerase Chain Reaction ; RNA, Messenger ; genetics ; metabolism ; Rats ; Recombination, Genetic ; Stem Cells ; cytology ; metabolism ; Transfection

Objective To test the changes of the potassium(K+)and magnesium(Mg2+)concentrations in vitreous humor of rabbits along with postmortem interval(PMI)under different temperatures, and explore the feasibility of PMI estimation using mixed-effect model. Methods After sacrifice, rabbit carcasses were preserved at 5℃, 15℃, 25℃ and 35℃, and 80-100μL of vitreous humor was collected by the double-eye alternating micro-sampling method at every 12 h. The concentrations of K+and Mg2+in vitreous humor were measured by a biochemical-immune analyser. The mixed-effect model was used to perform analy-sis and fitting, and established the equations for PMI estimation. The data detected from the samples that were stoned at 10℃, 20℃ and 30℃ with 20, 40 and 65 h were used to validate the equations of PMI estimation. Results The concentrations of K+and Mg2+[f(x,y)] in vitreous humor of rabbits under different temperature increased along with PMI(x). The relative equations of K+and Mg2+concentration with PMI and temperature under 5℃~35℃ were fK+(x,y)=3.413 0+0.309 2 x+0.337 6 y+0.010 83 xy-0.002 47 x2 (P<0.000 1), and fMg2+(x,y)=0.745 6+0.006 432 x+0.033 8 y(P<0.000 1), respectively. It was proved that the time of deviation for PMI estimation by K+and Mg2+was in 10 h when PMI was between 0 to 40 h, and the time of deviation was in 21 h when PMI was between 40 to 65 h. Conclusion In the ambient temperature range of 5℃-35℃, the mixed-effect model based on temperature and vitreous humor sub-stance concentrations can provide a new method for the practical application of vitreous humor chemi-cals for PMI estimation.

Objective To test the spatial resolution and average glandular dose (AGD) of full field digital mammography system to obtain high-resolution and contrast breast X-ray images with the lowest dose.Methods The M12 breast X-ray performance test phantom was placed on the breast support center position,then line-pair card was put on the M12 phantom and fixed groove,and exposure was carried out with auto exposure control,28 kV tube voltage and common anode filtration combination so as to detect the spatial resolution.Half-value layer (HVL) was measured by Fluke TNT12000 ray detection tools,2 mm-thickness plate and aluminum sheets with different thicknesses,and then AGD was calculated accordingly.Results The spatial resolutions were all higher than 7 lp/mm and proved to meet the quality standard,which had the vertical spatial resolution being 8 lp/mm,the maximum value of the lateral spatial resolution being 10 lp/mm and the minimum value being 8 lp/mm.AGD was lower than 2 mGy and then proved qualified which was limited within 0.60 and 0.61 mGy.Conclusion The test of spatial resolution and AGD of full digital mammography system can assess the performance and radiation dose,reflect the performance of equipment,ensure image quality and lower radiation dose.

Objective To detect the SNR and geometric accuracy of MRI based on American College of Radiology (ACR)standards.Methods The SNR and geometric accuracy of Siemens Skyra 3.0T,Siemens Trio 3.0T and GE Excite HD 1.5T MRI were measured with ACR phantom,and the detection results were calculated according to the standards.Results The SNR values of Siemens Skyra 3.0T,Siemens Trio 3.0T and GE Excite HD 1.5T MRI were 589.98,438.50 and 277.12 respectively.Siemens Skyra 3.0T MRI had the values of geometric accuracy being-1.93％ at X direction,-3.20％ at Y direction and 0.68％ at Z direction,Siemens Trio 3.0T MRI had the value of geometric accuracy being-0.87％ at X direction,-2.33％ at Y direction and 1.49％ at Z direction,GE Excite HD 1.5T MRI had the values of geometric accuracy being 0.20％ at X direction,-1.53％ at Y direction and 1.69％ at Z direction.Conclusion The detection of the SNR and geometric accuracy of MRI can effectively guarantee the image quality.

Objective: To investigate the clinical significance of tumor associated macrophages (TAM) in multiple myeloma (MM) and the relationship with angiogenesis and immunosuppression. Methods: Seventy cases of MM patients diagnosed from August 2015 to June 2017 were enrolled in the study as experimental group, 20 cases of benign hematological diseases (13 with iron deficiency anemia and 7 with megaloblastic anemia) patients as control group. Immunohistochemical method was used to detect the expression of CD163, CD34 and VEGF in bone marrow samples, and flow cytometry was used to detect the proportion of regulatory T cell (Treg cells), ELISA was used to detect the level of IL-10, and the clinical features were analyzed. Results: ①Among the 70 patients, there were 31 males and 39 females with a median age of 65 (50~78) years old. TAM infiltration density, microvascular density (MVD), VEGF expression level, Treg ratio and IL-10 level in bone marrow samples of 70 MM patients were significantly higher than those of benign hematological diseases (P<0.05). ②In the MM group, the above indexes of the patients with disease stabilized (15 cases) were lower than those of the newly diagnosed group (35 cases) and the relapse refractory group (20 cases) (P<0.05), those of relapse refractory group were higher than those of newly diagnosed group (P>0.05). ③Of the 35 newly diagnosed MM patients, 27 completed 4 courses of treatment. In the effective group (15 cases), the TAM infiltration density after treatment was significantly lower than that before treatment, the difference was statistically significant[(20.20±7.66) vs (28.87±11.97), t=2.362, P=0.025]; while in the ineffective group of 12 cases, the difference of the TAM infiltration density before and after treatment was not statistically significant[(42.00±13.76) vs (48.25±13.59), t=1.119, P=0.275]. ④TAM infiltration density in the effective group after bortezomib treatment (21 cases) were lower than those in the non-bortezomib treatment group (18 cases)[(16.52 ±4.26) vs (19.27 ±5.82), t=1.662, P=0.170]. ⑤The TAM infiltration density in MM patients was positively correlated with MVD, VEGF expression level, Treg cell ratio and IL-10 level (P<0.001). Conclusion: The infiltration of TAM in the microenvironment of MM, which may promoting angiogenesis and inhibiting immune response, is related to the occurrence, development, therapeutic effect and drug resistance of MM.
Aged ; Female ; Humans ; Macrophages ; Male ; Middle Aged ; Multiple Myeloma ; Neoplasm Recurrence, Local ; Neovascularization, Pathologic

In this study, we investigated the mechanism of crude extract of Psammosilene tunicoides(CEPT) in the treatment of rheumatoid arthritis(RA) based on the Nod-like receptor protein 3(NLRP3) inflammasome. The collagen-induced arthritis(CIA) mouse model was established. On day 32 after the primary immunization, according to the arthritis score, the mice were randomly divided into model group, positive control(methotrexate) group, low-and high-dose CEPT groups, and normal group, with 10 mice in each group. According to the administration dose of each group, the mice were continuously administered for 21 days. Every four days during the administration, the paw edema degree, arthritis score, and spleen index of the mice were measured; histopathological examination was performed for the ankles of the mice; the contents of IL-1β and IL-18 in the serum were determined; the protein expression levels of NLRP3, caspase-1, and apoptosis-associated speck-like protein containing a CARD(ASC), as well as the mRNA expression levels of NLRP3 and caspase-1 in the ankle joints of the mice were detected. The results showed that compared with those in the model group, the mice in the positive control group and CEPT groups had significantly decreased the contents of IL-1β and IL-18 in the serum and spleen index(P<0.01), significantly lowered arthritis score and degree of paw edema(P<0.01), alleviated arthritic infiltration of the knee, and down-regulated protein and mRNA levels of NLRP3, ASC, and caspase-1 in the ankle joint(P<0.01). These results suggest that P. tunicoides may reduce the paw edema and arthritis score and alleviate the inflammatory response in CIA mice by inhibiting the expression of NLRP3. This study provides a basis for the study of immune regulation of P. tunicoides in RA.
Animals ; Arthritis, Experimental/genetics* ; Arthritis, Rheumatoid/genetics* ; Caspase 1/genetics* ; Inflammasomes/genetics* ; Mice ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics*

Psammosilene tunicoides is one of the main ingredients of the "Yunnan Baiyao". P. tunicoides is an endangered species included in the secondary protection list in China Plant Red Data Book as well as the endemic species in Southwest China. Its natural resources could not meet the needs of pharmaceutical production. Construction of core collection of P. tunicoides will lay the foundation for germplasm improvement and molecular breeding. The sequence variation of the key enzymes gene locus (β-AS) were carried out to survey the population structure and population history of the species. Among the 11 populations across its geographical range, 36 haplotypes were identified. The levels of haplotype diversity (Hd=0.905) were high, while the levels of population differentiation (GST=0.280) were low. Analysisof molecular variance (AMOVA) indicated that a significantly greater proportion of total genetic variationpartitioned among populations thanwithin populations (values of 77.43% and 22.57%, respectively). These results in combination with the star-like phylogenetic network analysis indicate that Hap1 as an ancestral haplotypewas shared in four populations, Hap2, Hap4, Hap15 and Hap16 are occurred in two populations, the remains as private haplotype only distributed in single population. The strategy of core collection was constructed in order to maximumpreserve genetic diversity of P. tunicoides.
Caryophyllaceae ; genetics ; China ; Genetic Variation ; Genetics, Population ; Haplotypes ; Phylogeny ; Plants, Medicinal ; genetics