Objective To construct the eukaryotic expression vector of linker for activation of T cells(LAT) gene in mouse. MethodsLAT cDNA of mouse mononuclear cells was amplified with polymerase chain reaction.The fragment was inserted into the eukaryotic expression vector of pCMV-HA plasmid.The recombinant plasmid was verified by DNA sequencing and used to transfect lymphocytes of the asthmatic model. Results The length of amplified fragment was 729 bp.The sequence of LAT gene was confirmed by Genbank.The LAT protein could be detected in the lymphocytes of asthmatic model.Conclusion The recombinant eukaryotic expression vector pCMV-HA-LAT can be successfully constructed.

Argentina ; epidemiology ; China ; epidemiology ; Cross-Sectional Studies ; Hemophilia A ; diagnosis ; epidemiology ; therapy ; Humans ; Prognosis ; Russia ; epidemiology ; Surveys and Questionnaires ; United States ; epidemiology ; Urban Population

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast ; pathology ; Breast Neoplasms ; drug therapy ; pathology ; Cyclophosphamide ; therapeutic use ; Diagnosis, Differential ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Lymphoma, B-Cell ; drug therapy ; pathology ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; pathology ; Male ; Middle Aged ; Prednisone ; therapeutic use ; Sarcoma ; pathology ; Spleen ; pathology ; Splenic Neoplasms ; drug therapy ; pathology ; Vincristine ; therapeutic use

This paper is aimed to provide the methods of quality control and bioassay of traditional Chinese medicine injections including bioassay method. Shuanghuanglian freeze-dried powder for injection (SFPI) was chosen as study object. HPLC-ELSD fingerprints of SFPI had been established and the samples were differentiated by similarity calculation. Meanwhile, biological profiles of SFPI on Escherichia coli had been established by microcalorimetry. The similarity values were calculated using the correlation coefficient, based on quantitative thermo-kinetic parameters (T2m, Tj, I%). The results indicated that HPLC-ELSD fingerprints, which showed content changes of chemical components, could not monitor minimal variation of different samples, especially that of biological pollutants, while biological profiles could sensitively detect antibiotic activity alterations of the samples, which were kept under specific conditions. In conclusion, characterized by two-dimension, microcalorimetry could supply thermograms as biological profiles characterized to describe the bioactivity of drugs. This study could clearly demonstrate that the correlative detection was proposed as an efficient strategy for quality control of SFPI, based on HPLC-ELSD fingerprints and biological profiles, which could detect quality fluctuation of samples early and quickly and predict the potential adverse drug events (ADE) for ensuring clinical safety.

In this study, microcalorimetry was adopted to establish a novel method for detecting the hemagglutination process of Radix Isatidis (Banlangen in Chinese, BLG), and to evaluate the hemagglutination activity diversity of BLG from various habitats. The hemagglutination biothermokinetics curves of positive reagent (phytohemagglutinin, PHA) and 8 batches BLG from different regions of the hemagglutination with 20% rabbit erythrocyte were recorded by microcalorimetry, then biothermokinetics parameters were abstracted, the hemagglutination utility of samples were calculated and analyzed by principal component analysis (PCA) and cluster analysis (CA), meanwhile the results were authenticated by micro-plate agglutination. It showed that the hemagglutination was an exothermic reaction, the reaction rate constant (k: 0.039-73.6 min(-1)), maximum reaction power (Pmax: -1 140.2 - 988.2 microW) and reaction enthalpy (Hi: -529.9 - 717.9 microJ) had good linear correlation with BLG extraction concentration (0.2-1.0 g mL(-1), r > 0.97), and PCA showed Pmax (531-1 335 microW) and Ht (585.2-989.2 microJ) could represent the hemagglutination activity diversity of BLG samples, just confirming with the results of micro-plate agglutination (the agglutination dilution was 3-11 respectively). According to the hemagglutination utility, the BLG samples from Good Agriculture Practice (GAP) regions, main producing area and general regions could be clustered correctly; meanwhile, the biothermokinetics curves with perfect distinctive fingerprint and specificity could give out more information for the quality control and evaluation for BLG. In conclusion, the microcalorimetry method established for detecting the hemagglutination activity of BLG samples on rabbit erythrocyte is sensitive and reliable, and could be adopted as an effective technique in detection aggulatination precisely, quantitatively and consecutively; and provide a novel approach for examining and evaluating quality for Chinese herbal medicine with aggulatinative activity such as BLG.

The paper is to report the establishment of a method for quickly evaluating compatibility of Chinese medicines injections. Isothermal titration calorimetry (ITC) was used to evaluate compatibility of Yiqifumai (YQFM) and vitamin C injection (Vc)/5% glucose injection (5% GS). The diversification of Gibbs free energy (deltaG), enthalpy (deltaH) and entropy (deltaS) were used to decide reaction types of colliquefaction procedures of different injections. The reactive profiles were used to determine signs and quantity of heat. And high performance liquid chromatography (HPLC) was used as a supportive method for ITC. Then, feasible binding sites were analyzed based on the information of spatial structures of major compositions. During the colliquefaction procedure of YQFM and Vc, [deltaH] > T[deltaS], so, the reaction is enthalpy-driving. And the reactive profile showed that a big deal of heat was given out during the procedure. Obviously, chemical reactions happened and the major compositions changed. On the other side, the reaction of YQFM mixed with 5% GS was entropy-driving, because [deltaH] < T[deltaS]. The reactive profile showed there was a little heat gave out. So, non-chemical reactions happened, and the major compositions did not change. The conformity existed between the results of ITC and HPLC. ITC could be used to evaluate the compatibility of Chinese medicines injections because of the advantages of ITC, such as real time, fast, sensitive and having more parameters.

· AIM: To inspect and compare the functional vision of an aspheric intraocular lens (Tecnis) with those of conventional monofocal silicone and acrylic intraocular lens and multifocal intraocular lens (Array).· METHODS: The IOLs were tested in the eye model, which was designed to be optically equivalent to the theoretical eye model. The eye model is a combination of a spherical photographic lens with 35mm focal length ( IOL put in a water cell)and a charge coupled device (CCD) camera. The images constructed by the lenses are observed on a monitor of personal computer and the contrasts of the images are analyzed by using commercial image processing software.· RESULTS: The modulation transfer function of the eye model equals the scale produced by the theoretical eye model. The images constructed by changing the diameter of aperture stop and IOL.· CONCLUSION: The proposed eye model is useful for testing functional vision and for inspecting differences of intraocular lens.

Objective To investigate the effect of a fast-track based clinical nursing pathway of hepatic carcinoma on patient satisfaction and job satisfaction of nurses. Methods A total of 91 primary hepatic carcinoma (PHC) patients undergoing elective hepatectomy were enrolled and were divided into two groups by admission time, namely the control group (48 cases) and the experimental group (43 cases). The patients in the control group received traditional perioperative nursing care of hepatectomy, while the cases in the experimental group participated in fast-track nursing pathway. Patient satisfaction and nurse job satisfaction were measured before and after implementation. Results Final analysis included 45 patients in the control group and 40 patients in the experimental group. In the experimental group, the total score of patient satisfaction and the scores of all dimensions (including services and techniques, concern and care, services and techniques and overall satisfaction) were (132.75 ± 13.71), (48.30 ± 6.53), (42.60 ± 4.62), (35.98 ± 3.85), (5.88 ± 0.94), while they were (122.89 ± 16.62), (44.53 ± 7.74), (39.49 ± 5.67), (33.53±4.40), (5.33±1.13) in the control group. And the scores of the experimental group were significantly higher than that of the control group (t=-2.961, t=-2.409, t=-2.753, t=-2.709, t=-2.389, P<0.05). Furthermore,after implementing the fast-track clinical nursing pathway,the total score of nurse job satisfaction and the scores of six factors were (143.23 ± 9.40), (28.45 ± 2.72), (24.82 ± 2.28), (23.05 ± 1.33), (13.91 ± 1.95), (15.05 ± 1.43), (16.64 ± 1.65) while they were (127.36 ± 13.14), (25.68 ± 4.16), (19.64 ± 2.75), (20.23 ± 2.35), (13.23 ± 2.22), (13.68 ± 2.01), (14.50 ± 1.95) before the implementation. The former was significantly better than the latter (t =-12.462,-7.743,-12.670,-6.560,-2.417,-1.483,-6.587,P<0.05). However, no differences were observed in the other two dimensions of wages and benefits andwork-family balance(t =-1.945,-1.312,P>0.05) since the scores after implementation were (13.86 ± 2.12), (7.45 ± 1.57), while that of (13.18 ± 2.30)、(7.23 ± 1.41) were obtained before performance. Conclusions Application of fast- track nursing pathway in perioperative patients with PHC can contribute to promoting the level of patient satisfaction and job satisfaction of nurses and improving nursing quality. It is suggested that this nursing pathway will have more potential value for PHC patients.

OBJECTIVETo study the ability of bone marrow mesenchymal stem cells (BMSCs) to repair the internal environment of the testis in male azoospermia rats.

METHODSWe established azoospermia models in 22 six-week-old male SD rats by intraperitoneal injection of busulfan at 20 mg per kg body weight. We transplanted allogeneic rat BMSCs (rBMSCs) into the testicular seminiferous tubules of the model rats and, 30 days after transplantation, observed the composition and structure of the seminiferous tubular cells by HE staining and detected the expressions of CD44, CD106, and c-kit in the rBMSCs by immunohistochemistry.

RESULTSThe number of epididymal sperm was significantly reduced in the model rats as compared with the normal controls (P < 0.01). CD44 and CD106, but not c-kit, were expressed in the isolated rBMSCs. At 30 days after transplantation of rBMSCs, lots of new cells were observed in the seminiferous tubules, some expressing CD106 and some expressing the germ cell surface marker c-kit.

CONCLUSIONBMSCs can transdifferentiate into germ cells and repair the damaged seminiferous tubules of sterile rats.

Animals ; Azoospermia ; chemically induced ; therapy ; Biomarkers ; metabolism ; Bone Marrow Cells ; Busulfan ; Cell Membrane ; metabolism ; Epididymis ; Hyaluronan Receptors ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Proto-Oncogene Proteins c-kit ; metabolism ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; anatomy & histology ; metabolism ; Spermatozoa ; Staining and Labeling ; Vascular Cell Adhesion Molecule-1 ; metabolism

Objective To investigate the expression of miR-21 and PDCD4mRNA in colorectal cancer and their correlation with clinicopathological characteristics,to elucidate relationship between PDCD4and miR-21 in vivo.Methods Expression of RNA (including miR-21and PDCD4mRNA) and PDCD4 protein were detected respectively by quantitative real-time reverse transcriptase-PCR and SP immunohistochemical staining in 43 cases of colorectal carcinoma and corresponding normal mucous membrane tissue.Results In colorectal carcinoma,expression of miR-21 was higher than that of the control ( P ＜ 0.05 ),and expression of PDCD4mRNA lower than the control ( P ＜ 0.05 ).Expression of miR21 was associated with lymph node metastasis and clinical stages ( Ⅰ + Ⅱ,Ⅲ + Ⅳ ) ( P ＜ 0.05 ).On the other hand,no significant differences were observed regarding sex,tumor site,size,local invasion,distant metastases,clinical stages( Ⅰ,Ⅱ ) (P ＞ 0.05 ).Expression of PDCD4mRNA was associated with local invasion,clinical stages ( Ⅰ,Ⅱ ) ( P ＜ 0.05 ).While no significant differences were found concerning sex,site,size,lymph node metastasis,distant metastases,clinical stages( Ⅰ + Ⅱ,Ⅲ + Ⅳ ) ( P ＞ 0.05 ).miR21 levels was negatively correlated with PDCD4 expression including nuclear and nuclear and cytoplasmic put together (P ＜ 0.05 ),in contrast to PDCD4mRNA and PDCD4 expression in cytoplasmic ( P ＞ 0.05 ).Conclusions ( 1 ) Abnormal expression of PDCD4 mRNA and miR-21 correlate with prognosis in colorectal cancer.(2) miR-21 suppress translation of PDCD4mRNA by binding to the 3'-untranslated region (3'-UTR)of target PDCD4mRNA.