Objective To investigate the effects of staphylococcal enterotoxin A SEA gene on target cells mediated by replicative-deficient recombinant adenovirus vector. Methods Lymphocytes of C57BL/6 mice were infected with various titers of recombinant adenoviruses. Supernatants were collected after 12 h, 24 h, 48 h, 72 h, 96 h, 120 h and 144 h of incubation and analyzed for proliferation of lymphocytes by MTT assay. IL-2 level in the culture supernatants was measured with ELISA. The killing effect of lymphocytes was also observed by MTT assay. Results Proliferation response and elevated levels of IL-2 were observed in experimental group. The killing effect on B16 cells was stronger in experimental group, which seemed to be dose-dependent with the increase of ratio of lymphocytes/target cells. Conclusions SEA gene can be expressed in lymphocytes of C57BL/6 mice mediated by replicative-deficient recombinant adenovirus vector. The expressing products can activate lymphocytes of C57BL/6 mice, which kill B16 cells in vitro.

Objective To explore the clinical and pathologic features of Langerhans cell histiocytosis(LCH).Method Thirty-eight cases of LCH from both in hospital and outpatient department in dermatology and pediatry department from Jan.1991 to Jun.2006 were analyzed about the features of clinic,pathology and immunohistochemistry.Results The mean age of onset was 2.18 years old.The male /female ratio was 1.71.Skin lesions occurred in 78.9% of the patients.Among them,68.5% were as the first manifestation.The eruptions mainly distributed on trunk,90% of them presented as hemorrhagic maculopapules,nevertheless,3.3% of the eruptions showed as crust with hilar depression,which was similar to pityriasis lichenoides etvarioliformis acuta.Fever,hepatomegalia and splenomegia occurred in patients at a rate of 60.5%,68.4%,55.3% respectively.Thirty-one point six percent of the patients had got lymphadenectasis,the neck and inguinal lymph nodes were the common site to be affected.Ossature involvement occurred in 31.6% of the patients,which 8.3% got multiple injuries,howe-ver,91.7% got a solitary bone involved.Skull was the main site to be injured,else were lumbar,humerus,hipbone,an so on.Respiratory tract,auditory canal,mucosa were also the sites involved in this disease,but the incidence rate was lower than 10%,respectively.The laboratory data showed that 81.3% of the patients were anaemia,60.7% with abnormal subgroup of T-cell,and 32.1% positive for EBV-IgG.The skin histopathology data of 26/30 cases revealed that lichenoid infiltrates of Langerhans cells confined to the upper dermis.Cytologic features were cells with abundant eosinophilic cytoplasms,and longitudinally grooved or reniform nuclei.Lymphocytes and a few eosinophils also could be seen.Four cases of thirty showed that the proliferative Langerhans cells were with pale cytoplasms,besides,there were numerous eosinophils,and sometimes a few multinucleate cells were scattered.The immunity test of 20 cases of thirty displayed that CD1a(+)S100(+)KP-1(-).Biopsy of lymphaden and tumor of the skull of the rest 8 patients were all diagnosed as eosinophilic granuloma through both hematoxylin and eosin-stained section and immune marks.Conclusions Multiple systems can be involved in LCH.Hemorrhagic maculopapules,fever and splenohepatomegalia are common presentations in this disease.The morphous of nucles of histiocytes is particular,and to diagnose definitely,both CD1a(+) and S-100(+) are needed.

The arrangement of fascicles of musculocutaneous nerve at different levels in brachial plexus were studied in 22 adult cadavers. The nerve fascicles were dissociated and traced under operative microscope. At the distal end of the musculocutaneous nerve, the muscular and cutaneous fascicles are separated.The muscular fascicles are situated at the latero-anterior quadrant of the cross-section of the nerve.The mixed fascicle are become predominated proximally. The muscular and the muscular predominated mixed fascicles are still situated at latero-anterior quadrant. At the level 1 cm below the clevicle, 1cm and 1.5 cm above the clevicle,the fascicles of the nerve are situated at the lateral superio-anterior quadrant of the crosssection of relative parts of brachial plexus. The fascicles of the nerve are situated at the anterior quadrant of the crosssection of superior trunks. The fibers of musculocutaneous nerve are originate from C5, C6 and C7, being 31.6,％, 64.6％ and 3.8％,respectively.

This paper briefly analyzed and discussed the current status and major scientific challenges of traditional Chinese medicine (TCM) formulaology research. To promote formulaology research, a new strategy and corresponding technology, network formulaology, were proposed to reveal the complex interaction between functional chemome and biological responses network. The research framework and directions of network formulaology were also summarized and prospected.
Chemistry, Pharmaceutical ; methods ; standards ; Drugs, Chinese Herbal ; chemistry ; Internet ; Medicine, Chinese Traditional ; standards

Objective To analyze the dietary habits, energy intake and expenditure, anthropometrics, and body composition of the outpatients visiting the weight loss clinic of Beijing Hospital.Methods We pro-spectively enrolled 89 consecutive patients with body mass index ( BMI) ≥24 kg/m2 from November 2014 to August 2015 in the weight loss clinic of Beijing Hospital.There were 35 male and 54 female, with the mean age of (45.8 ±16.4) years.We divided them into two groups:the diabetes group (n=35) and the non-diabetes group (n=54), and compared the dietary habits, energy intake and expenditure, anthropometrics and body composition between the two groups.Results Regardless of diabetes, the overweight and obese patients all ate fast, mostly finishing a meal in about 10 minutes.They preferred Chinese food and meat, and disliked hot food.The frequency of dinning out in the non-diabetes group (3-5 times per week) was higher than that in the diabetes group (1-2 times per week) .Compared with the diabetes group, the non-diabetes group had higher fat-to-energy ratio [(34.9 ±7.6)%vs.(30.8 ±5.9)%], but lower carbohydrate intake [(232.2 ±59.7) g vs.(283.6 ±89.5) g], carbohydrate-to-energy ratio [ (47.9 ±8.3)%vs.(53.4 ±7.1)%], and the ratio of resting metabolic rate to body weight [ (66.9 ±9.6) kJ/(d? kg) vs.(71.1 ±7.9) kJ/(d? kg)] (all P<0.05).There were no statistically significant differences between the two groups in total energy intake, pro-tein intake, high quality protein intake, fat intake, protein-to-energy ratio, and resting metabolic rate (all P>0.05).Anthropometrics showed that the mean BMI of the patients was (32.8 ±4.4) kg/m2, with the maxi-mum being 53.5 kg/m2.The hip circumference [ (117.15 ±9.9) cm vs.(111.1 ±8.2) cm], upper arm circumference [ (36.4 ±3.8) cm vs.(34.0 ±3.3) cm], and triceps skinfold thickness [ (36.1 ±8.9) mm vs.(31.6 ±8.8) mm] were larger in the non-diabetes group than in the diabetes group (all P<0.05), but the mean age was lower in the non-diabetes group [ (41.7 ±16.9) years vs.(52.9 ±13.1) years) (P=0.01).There were no statistically significant differences between the two groups in body weight, BMI, waist circumference, neck circumference, and bilateral hand grip strength (all P>0.05).According to body compo-sition analysis, the body weight [ (94.8 ±18.3) kg vs.(86.9 ±17.2) kg], body fat mass [ (39.7 ± 11.3) kg vs.(33.5 ±8.9) kg], body fat percentage [ (41.7 ±6.5)%vs.(38.5 ±6.7)%], and visceral fat area [ (145.3 ±24.8) cm2 vs.(130.7 ±27.5) cm2 ] were larger in the non-diabetes group than in the di-abetes group ( all P<0.05) .There were no statistically significant differences between the two groups in BMI and skeletal muscle mass (both P>0.05).Conclusion Compared with diabetes patients, overweight and obese non-diabetes patients may be younger, having worse dietary habits, and having larger body fat mass, body fat percentage, and visceral fat area.

Four salts of ticagrelor, ticagrelor-3,5-dinitrobenzoic acid, ticagrelor-pyrazinamide, ticagrelor-D-proline and ticagrelor-L-proline were prepared by solvent suspension and liquid-assisted grinding to improve the solubility of ticagrelor. The compounds were characterized by powder X-ray diffraction, Fourier transform infrared spectroscopy, differential scanning calorimetry, nuclear magnetic resonance spectroscopy, elemental analysis, and the intermolecular salt-bonding forces were analyzed. The equilibrium solubility of salts and pure drug in hydrochloride buffer pH 1.2 and phosphate buffer pH 6.8 were measured by high-performance liquid chromatography. Ticagrelor was salted with 3,5-dinitrobenzoic acid, pyrazinamide, D-proline, L-proline all in a stoichiometric ratio of 1∶1; with the exception of ticagrelor-D-proline, the solubility of the other three salts provided significantly improved solubility in hydrochloride buffer pH 1.2, and the equilibrium solubility of ticagrelor-3,5-dinitrobenzoic acid was increased by approximately 1.7 folds as compared to pure drug. Salt-forming technology is convenient and can improve the solubility of ticagrelor.

Abscess ; therapy ; Adult ; Aged ; Catheterization ; methods ; Drainage ; methods ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed

Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Cryptotanshinone (CPT), a lipid soluble active compound in Salvia miltiorrhiza, has a significant inhibitory effect on multiple malignant tumors, e. g. chronic myeloid leukemia (CML) cells and can effectively enhance imatinib's chemotherapeutic effect. However, its functional molecular mechanism remained unclear. In this experiment, the authors conducted a systematic study on the effect of CPT on the imatinib sensitivity and P-glycoprotein (P-gp) expression in CML cells by using CML cells K562 and imatinib persister K562-R. The MTT assays were performed to determine CPT's impact on the inhibitory effect of imatinib. Annexin V-FITC/PI staining analysis was used to detect the changes in the cell apoptosis rate. The active changes in apoptosis regulatory proteins Caspase-3, Caspase-9 and PARP were determined by Western blot. After the cells were pretreated with the gradient concentration of CPT, the expression of P-gp was analyzed by Western blot and flow cytometry. The changes in intracellular concentrations of imatinib were determined by HPLC analysis. The results indicated that the pretreatment with CPT significantly increased the proliferation inhibiting and apoptosis inducing effects of imatinib on K562 and K562-R cells as well as the degradation product expression of pro-apoptotic proteins Caspase-3, Caspase-9 and PARP, with a significant difference with the control group (P < 0.01). However, CPT showed no impact on the P-gp expression in CML cells and the intracellular concentrations of imatinib. In summary, the findings suggested that CPT enhanced the sensitivity of CML cells to imatinib. Its mechanism is not dependent on the inhibition in P-gp expression and the increase in intracellular drug concentration.
ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Caspase 9 ; genetics ; metabolism ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Imatinib Mesylate ; pharmacology ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; genetics ; metabolism ; physiopathology ; Phenanthrenes ; pharmacology

BACKGROUND:A group of nuclease-like proteins were previously purified from Eisenia foetida tissues, exploring primary structures of these proteins wil help to uncover basic structure characteristics of them and provide foundations for the study addressing the relationship of their structures and functions.
OBJECTIVE:To explore primary structures of nuclease-like proteins EWD1 and EWD2.
METHODS:Edman degradation method was used to sequence the N-terminal amino acids of EWD1 and EWD2, acid hydrolisis method was used to analyze amino acid compositions of EWD1 and EWD2, LC-MS/MS was used to analyze some peptide sequences within the proteins, and MALDI-TOF-MS was used to calculate the number of the disulfide bonds and the contents of polysaccharides.
RESULTS AND CONCLUSION:Among the amino acid compositions in EWD1 and EWD2, the sum contents of aspartate and asparagines were the highest (al nearly 10%), the contents of hydrophobic amino acids were also high, and the contents of cysteine was low. The EWD1 and EWD2 had similar amino acid compositions with other nucleases. Edman degradation results showed that, the N-terminal sequences of the large subunit of EWD1 were in turn as fol ows:D, E, W, V, Y, P;the N-terminal sequences of EWD2 were as fol ows:L, L, G, P, Y, K, P, K, C. The results of LC-MS/MS indicated the two proteins were novel proteins;MALDI-TOF-MS results showed that 8 cysteine residues formed 4 disulfide bonds in EWD1, 6 cysteine residues formed 3 disulfide bonds in EWD2. EWD1 and EWD2 were al glycoproteins, the content of polysaccharides was 17.3%in EWD1 and 15.6%in EWD2.