Based on avian leukosis virus ( ALV) p19 gene terminal nucleotide sequence, a 82 bp double-stranded DNA fragment was chemically synthesized and cloned into the expression vector pGEMEX-1. The sequencing result indicated th at the cloned fragment was a correct version of the one designed both in nucleot ide sequence and in its open reading frame. The recombinant was used to transfor m E.coli BL21 (DE3). The cloned fragment was expressed as a fused protein wi th T7 gene 10 leader peptide and was shown to be 34 kD in size on SDS-PAGE gel when induced with 1 mmol/L IPTG. The expression product was able to bind immunol ogically to rabbit anti-ALV serum in Western-blot assay and is being tested to differentiate exogenous from endogenous ALV.

Objective To explore the corresponding relationship between isolated Q waves in lead Ⅲ on the electrocardiography(ECG) and echocardiographie segmental diagnosis of myocardial infarction(MI).Methods The shape magnitude of Q waves of 27 patients with MI and the relationship with eehocardiographie features were investigated. Results Twenty-four patients with inferior wall myocardial infarction had close relationship with isolated Q waves in lead Ⅲ on the ECG. Conclusions There is definite corresponding relation between isolated Q waves in lead Ⅲ and inferior wall myocardial infarction.

In order to solve the problems of erratic drug absorption and low bioavailability after oral administration for poorly-water soluble drugs due to low solubility, a series of novel pharmaceutical dosage forms as solid dispersion, liposome, microemulsion, vesicle, cyclodextrin inclusion complexes and drug nanocrystal have been developed in recent years. Among which drug nanocrystal attracts more attentions for its simpler preparation method, higher drug loading and easier manufacturing technology in the design of dosage forms suitable for different administration routes. In this paper, the nanocrystals of the poorly-water soluble drugs prepared based on bottom-up and top-down technologies were introduced. The characteristics and applications of the nanocrystal-based dosage forms as suspension, tablet and capsule were also introduced and carefully evaluated with the focus on their pharmacokinetics, pharmacodynamics and tissue targeted drug distribution after delivery by oral administration, intravenous injection and pulmonary inhalation. The advantages of drug nanocrystals in their therapeutics effects over the bulk drugs were discussed together with the inherent mechanism. Finally, the problems existing in basic research and scaled-up manufacture of drug nanocrystal as well as the possible ways of solution were listed out so as to make the nanocrystal-based preparations exert their maximum therapeutic effect after clinical application.

Objective To investigate postoperative psychological experiences of patients underwent kidney transplant donated by his/her parent,and to discover mental signs and psychological journey in individual growth.Methods Semi-structured in-depth interviews were conducted among 17 patients underwent kidney transplant donated by his/her parent,and Giorgi analysis method was used to analyze data.Results The postoperative psychological experiences of patients mainly contained 4 aspects:posttraumatic growth,rebuilding meaning of life,being grateful for experience,coexisting of hope and challenge.Conclusion The patients have complex psychological experiences,having positive aspects derived from rehabilitation and improved quality of life,as well as negative aspects derived from concerning health of the donor,fear for complication and rejection,and lack of sound supporting system.Medical staff should guide patients to adjust cognitive and behavioral patterns and face life events directly,integrate medical/social support resources and enhance patients' psychological capital to face future life.

ObjectiveTo investigate the possible mechanisms for biocompatibility of chitosan material using agarose/chitosan blended hydrogels as a model.Methods A series of agarose/chitosan blended hydrogels with different chitosan content were prepared by the blending method.The chemical groups of the blended hydrogels were analyzed by the Fourier transform infrared (FTIR) spectroscopy.The blending compatibility between the agarose and chitosan was evaluated with the fluorescein-4-isothiocyanate (FITC) staining method.The charge of the blended hydrogels was determined by the zeta potential measurement.The adsorption of total fetal bovine serum (FBS) proteins and bovine serum albumin (BSA) on the blended hydrogels was measured by the bicinchoninic acid (BCA) method.The adsorption of fibronectin (FN) on the blended hydrogels was measured with ELISA.Cell culture experiment adopted human microvascular endothelial cell line (HMEC-1) as the model.The cytocompatibility was studied by evaluating adhesion,proliferation,and morphology of the cells on the blended hydrogels.Results Characteristic chemical groups of chitosan could be detected in the agarose/chitosan blended hydrogels.The chitosan had a good blending compatibility with the agarose.The amino groups of chitosan were uniformly distributed in the blended hydrogels.The blended hydrogels were strongly positively charged at acidic pH (pH 3.0),however,the zeta potentials of all the hydrogels were reduced to nearly 0 mV at neutral pH (pH 7.4).There were no significant differences in the adsorption of total FBS proteins and BSA between the blended hydrogel groups.However,the adsorption of FN on the hydrogels significantly increased with the increase of chitosan content.Cell culture experiment indicated that the cytocompatibihty of the blended hydrogels was significantly improved with the increase of chitosan content.The HMECs exhibited higher levels of adhesion,spreading,and proliferation on the hydrogels with higher chitosan content.ConclusionResults in this study indicated that the chitosan component preferentially adsorbed FN compared to the other serum proteins,leading to adhesion and spreading of the cells on the blended hydrogels.In contrast to prevailing views,it was found in the present study that the biocompatibility of chitosan did not relate to its positive charge.

Objectives To compare clinical manifestations and electrophysiological features in patients with chron?ic inflammatory demyelinating polyneuropathy (CIDP) and Type-I Charcot Marie Tooth Disease (CMT-I) for guiding dif?ferential diagnosis. Methods Data including clinical manifestations and electrophysiological indexes was collected from thirty-one CIDP cases and 28 CMT-I cases. Correlation analysis was used to assess the association of the severity of electrophysiology with the severity of clinical symptoms. Results There were statistically significant differences in onset site, sensory dysfunction, foot deformity and cerebrospinal fluid protein between these two groups (P<0.05). There were significant differences in indexes of nerve conduction and needle electromyography between these two groups (P<0.05). The severity of clinical symptoms was not related with the severity of electrophysiology in CMT-I group (r=0.27,P>0.05). Conclusions Differential diagnoses of CIDP and CMT-I can be made based on clinical manifestations and electro?physiological features.

A comparable study were carried out by determination of trace elements on five marine-derived shell traditional Chinese medicine (TCM) (Ostreae Concha, Haliotidis Concha, Margaritifera Concha, Meretricis Concha, and Arcae Concha), which were recorded in the Chinese Pharmacopoeia (2010 version). Seven trace elements in 51 batches of this type of shell TCM were analyzed by Inductively Coupled Plasma Mass Spectrometry (ICP-MS), combined with principal component analysis (PCA) methods. The content of element Se, which exhibited significant differences among different drugs, could be used as a key element to distinguish this type of drugs. Meanwhile, the contents of elements Co, Cu, Mo, and Ba in Haliotidis Concha, Co and As in Margaritifera Concha, Mo and As in Meretricis Concha, Mo, As, and Ba in Arcae Concha, and Zn in Meretricis Concha were relatively stable. In the PCA plot, Arcae Concha and Meretricis Concha could be efficiently distinguished from Ostreae Concha together with Haliotidis Concha, and Margaritifera Concha. The results also showed a correlation with their medicinal function. In conclusion, trace elements in marine-derived shell TCM could not be neglected for their quality control.
Animal Shells ; chemistry ; Animals ; Aquatic Organisms ; chemistry ; Bivalvia ; chemistry ; Mass Spectrometry ; Medicine, Chinese Traditional ; Trace Elements ; analysis

Objective To investigate the effect of high mobility group box chromosomal protein 1 (HMGB-1) on the proliferation and inflammatory phenotype of human fibroblast like synoviocytes (FLS).Methods FLSs were isolated from the synovial tissues of rheumatoid arthritis (RA) patients undergoing joint replacement surgery.All the experiments described here utilize the FLSs between the third and sixth passages.FLS were incubated with HMGB1 at (100,500,2 000 ng/ml) or interleukin (IL)-1β (0.5 ng/ml) or lipopolysaccharide (LPS) (100 ng/ml) alone or HMGB1/IL-1β complexes or HMGB1/LPS complexes.Cell proliferation assay were used by CCK-8,IL-6,IL-8 and matrix metalloproteinase-3 (MMP-3) in culture supernatants were measured using enzyme-linked immunosorbent assays.The measurement data were compared with single factor analysis of variance.Results Stimulation with all concentrations of HMGB1,IL-1β and LPS alone did not affect the cell proliferation of FLS.HMGB1/IL-1β complexes and HMGB1/LPS complexes did not affect the cell proliferation of FLS,neither (F=0.415.P=0.915).Except high concentration of HMGB1 (2 000 ng/ml) could significantly stimulate the secretion of IL-6 from FLS [(23.0±1.1) ng/ml],HMGB1,IL-1β and LPS alone did not affect the production of IL-6,IL-8 and MMP-3.However,HMGB1/IL-1β complexes and HMGB1/LPS complexes increased IL-6,IL-8 and MMP-3 production from FLS (F=97.804,117.383,70.179,P=0.000).Conclusion Neither HMGB1,IL-1β,LPS alone nor HMGB1/IL-1β complexes or HMGB1/LPS complexes affect the cell proliferation of FLS.HMGB1 in complex with LPS or IL-1β boost IL-6,IL-8 and MMP-3 production in synovial fibroblasts from RA patients.

Aged ; Biopsy, Needle ; Humans ; Liver ; pathology ; Male ; Peliosis Hepatis ; diagnosis ; pathology

Limulus Factor C, a serine protease zymogen from the amoebocytes of the limulus, has high affinity for endotoxin. When Factor C is activated by endotoxin, it hydrolyses artificial tripeptide substrate and measurable products are released, so it can be used as an alternative reagent for endotoxin analysis. Factor C gene of Tachypleus tridentatus was obtained through RT-PCR and the recombinant protein was expressed by Bac-to-Bac/BmNPV baculovirus expression system in silkworm larvae. The activity of Factor C was detected with diluted serum of silkworm larvae, and the sensitivity of endotoxin detected was 0.2 EU/mL when the serum was diluted at 1:500. The silkworm larvae expressed limulus Factor C could be used to develop a new low-cost endotoxin test reagent.
Animals ; Arthropod Proteins ; biosynthesis ; Baculoviridae ; Bombyx ; metabolism ; Enzyme Precursors ; biosynthesis ; Genetic Vectors ; Larva ; metabolism ; Recombinant Proteins ; biosynthesis ; Serine Endopeptidases ; biosynthesis