There are reports about the chemical compounds of Ciwujia herbs, but with no study report about the chemical material basis of Ciwujia injection (CWJI). In this study, LC-MS(n) and LC-Q-TOF-MS techniques were adopted for a qualitative analysis on phenylpropanoids in CWJI. The Ultmate XB-C18 column (4.6 mm x 250 mm, 5 microm) was adopted and eluted with the mobile phase of 0.5% formic acid-water and acetonitrile, with the flow rate at 0.8 mL x min(-1) and the column temperature at 20 degrees C. Based on the data of high-resolution and multi-stage MS, control products and literatures, altogether 54 phenylpropanoids were identified in Ciwujia Injection, including 34 phenylpropanoids, 16 ligans and 4 coumarins. Among them, 28 were reported for the first time in Ciwujia, and 14 compound structures were identified in comparison with the control products. The method established in this study could be used to simply and rapidly identify phenylpropanoids in CWJI. The findings provide scientific data for defining the chemical material basis of CWJI.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Eleutherococcus ; chemistry ; Mass Spectrometry ; methods ; Molecular Structure

Objective To investigate in vitro activity of antimicrobial agents against Enterococcus spp . isolated from clinic specimens in a hospital.Methods 188 Enterococcus spp . isolates from specimens sent by clinic depart-ments in June 2013-July 2014 were identified and performed antimicrobial susceptibility testing.Results Of 188 En-terococcus spp . isolates,119 were Enterococcus faecium (E.faecium),60 were E.faecalis ,and 9 were E.avium, these strains were mainly isolated from urine (34.57%)and blood specimens (19.15% ).No daptomycin and linezolid-resistant strain was detected;resistant rates of E.faecium to vancomycin was 1 .68%,to penicillin, ampicillin,high concentration gentamycin,erythromycin,and levofloxacin were all > 70%;except tetracycline, resistant rates of E.faecalis to the other antimicrobial agents were all lower than E.faecium,resistant rates of E. faecalis to penicillin and ampicillin were 16.67% and 13.33% respectively.Conclusion Daptomycin has high activity against Enterococcus spp . in this hospital.

Objective： The current study was designed to investigate the role of foods such as fish juice, pickles, bacon, salted fish, smoked food, fruit and vegetables in the etiology of esophageal cancer in Chaoshan region of China. Method： A large case-control study was conducted to investigate 1248 patients with squamous cell carcinoma of the esophagus and 1248 controls matched by sex and age. Results： After adjusting for the effects of occupation, areas of residence, income, alcohol, cigarette and tea, a strong association with a clear dose-response was observed between the frequencies of fish juice eating and esophageal cancer (P<0.01), and the relative risk for women was 23.5 and for men was 3.1, respectively. The results for pickles were similar to fish juice. A protective effect of fruits on esophageal cancer was observed both in women (OR=0.13) and in men (OR=0.46), and the risk was decreased (P< 0.01) as frequencies of fruits eating was increased. The risks were significantly higher in bacon and salted fish consumers than that in non-consumer, but the dose-response relationship was not significant. No significant effects, however, were detected in vegetables and smoked food consumption. Conclusions:To the authors' knowledge, this is the first report on relationship between the fish juice and high risk of esophageal cancer in population. Further epidemiological and experiment study are required to find a biological causal relationship between them.

Objective To observe the effect of Tangmaikang particles on insulin sensitivity in the treatment of type 2 diabetes.Methods A total of 120 patients with type 2 diabetes were selected, randomly divided into control group ( n=60) and treatment group ( n=60).Patients in control group received routine hypoglycemic, hypotensive and lipid adjusting treatment, while patients in treatment group were given Tangmaikang par-ticles 15 g? d-1 on the basis of conventional treatment.The treatment lasted for 3 months.The data of postprandial plasma sugar ( PPG) , gly-cosylated hemoglobin ( HbA1 c ) , fasting insulin ( FINS ) , insulin sensi-tivity ( HOMA-IS ) , low density lipoprotein ( LDL -C ) , triglyceride (TG), endothelin -1( ET-1) and the change of clinical symptoms in two groups before and after treatment were observed as well as the inci-dence of adverse reactions in 2 groups.Results The total effective rate was 88.33% in treatment group, better than control group (75.00%) .After treatment, the data of FINS, HOMA -IS, PPG, HbA1c were all significantly improved in two groups( P<0.05 ) , and treatment group improved more obviously than control group ( P<0.05 ) .Symptoms in two groups were both improved compared with before treatment( P<0.05) , and the treatment group was better than control group( P<0.05).But the TG level wasn′t significantly decreased in two groups( P>0.05).After treatment, LDL-C and ET-1 level in treatment group were significantly decreased ( P<0.05 ) , and the control group was not obvious (P>0.05).After the treatment for early diabetic nephropathy, ET -1 in treatment group was significantly lower (P<0.05), and the control group was not obvious (P>0.05).There was no adverse reactions found during treat-ment.Conclusion Tangmaikang particles are safe and effective for type 2 diabetes, which also can improve insulin sensibility, reduce insulin resistance and ET-1 level, especially can reduce the ET-1 level of patients with diabetic nephropathy.

A fragment of hepatitis E virus open reading frame-2(ORF2), located from amino acid residues 394 to 604, was expressed in E. coli. The recombinant protein NE2 was found to form homodimer mostly in SDS-PAGE, which can be dissociated to monomers when treated with urea, and it was recognized more strongly in its dimeric form than the monomer by HEV reactive human serum in Western blotting. Besides, many aggregated form of NE2 from dimer to at least hexamer can be seen in MALDI-TOF-MS. And when the hydrated dynamic semidiameter of NE2 moleculars in PBS was measured as about 4 nm by Dynamic Light Scattering (DLS), being equal to tetramer, but with high polydispersity, which suggested that the NE2 moleculars were existed in PBS in many different sizes. These results suggested that the recombinant NE2 can aggregate into several oligomer forms, the association in the dimer is most strong, and dimers can assemble further to form some super-structure.
Dimerization ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Gene Expression ; Protein Conformation ; Recombinant Proteins ; chemistry ; genetics ; metabolism ; Viral Proteins ; chemistry ; genetics ; metabolism

BACKGROUNDVibrio vulnificus (Vv) is an estuarine bacterium that can cause primary septicemia as well as serious wound infections. However, little is known about the mechanisms by which Vv infects dendritic cells (DCs) and its effects on cytoskeleton. In this study, we aimed to investigate the invasion, internalization, and the organelles damage of the cultured dendritic cells (a DC 2.4 strain) during Vv infection.

METHODSThe study model was the cultured DCs infected by a Vv 1.758 strain. Electron microscopy was used to observe the localization of bacteria at the different time points of infection, cell morphology, and the process of organelles changes. The cytoskeleton structure including the microfilaments and the microtubules rearrangement was examined under a fluorescence microscope.

RESULTSThe Vv were pinocytosised into the DC cells through double-sides, and localized at 1 - 2 mm of the inner side membrane. It took 1.3, 1.9, and 3.4 hours to reach the infection ratio of 25%, 50%, and 75%, respectively. Using electron microscopy, the DCs had been observed to have developed chromatin aggregation within 4.0 hours, and significant cytoskeleton structure disruption was noted within 6.0 hours.

CONCLUSIONThe high lethality of Vv infection may be associated with the direct disruption of the DCs cytoskeleton structure.

Animals ; Apoptosis ; physiology ; Cells, Cultured ; Cytoskeleton ; metabolism ; ultrastructure ; DNA Fragmentation ; Dendritic Cells ; metabolism ; microbiology ; ultrastructure ; Mice ; Microscopy, Electron ; Microscopy, Electron, Transmission ; Vibrio Infections ; metabolism ; Vibrio vulnificus ; pathogenicity

Animals ; Antioxidants ; pharmacology ; Apoptosis ; drug effects ; Atherosclerosis ; prevention & control ; Ceramides ; analysis ; Dietary Fats ; administration & dosage ; Emodin ; pharmacology ; Lipids ; blood ; Male ; Rabbits ; Signal Transduction ; Sphingomyelin Phosphodiesterase ; metabolism ; Sphingomyelins ; metabolism

OBJECTIVEChildhood absence epilepsy (CAE), a common form of idiopathic generalized epilepsy, accounts for 8% - 15% of all childhood epilepsies. A positive family history of epilepsy, a hereditary factor being one of the pathogeneses, is found in 15% - 44% of children with absence seizures. The phenotype of CAE is specific (including seizure forms and EEG), therefore it is suitable for genetic study. The purpose of this study was to confirm the linkage of childhood absence epilepsy to chromosome 8q24 in China.

METHODSTwenty-nine trios families (a patient and his/her parents) as patient group and 10 normal trios families as control group were investigated for chromosome 8q24 by haplotype analysis with 5 microsatellite DNA markers (D8S554, D8S534, D8S1100, D8S1783, D8S1753). Genomic DNA was isolated from 4 ml human peripheral blood by using the conventional procedure, and then was treated using the PCR method. PCR products were analyzed by gene scan. Statistical methodology included haplotype-based haplotype relative risk (HHRR) and transmission disequilibrium test (TDT).

RESULTSIn this study, the polymorphism information content (PIC) of 5 microsatellite DNA markers were: 0.519, 0.828, 0.528, 0.654 and 0.772. HHRR showed D8S554(4) (chi(2) = 5.939, P < 0.05), D8S1100(3) (chi(2) = 5.081, P < 0.05), D8S1783(6) (chi(2) = 4.308, P < 0.05). TDT showed D8S554(4) (chi(2) = 4.46, P < 0.05), D8S1783(6) (chi(2) = 4, P < 0.05). In order to exclude false association results, the authors analyzed every family in detail. Four trios families transmitted allele D8S1783(6) to their offspring, and the same allele hasn't been found in controls. The further work showed that locus D8S1783 had transmission disequilibrium with CAE, the other two loci were a false association.

CONCLUSION(1) Childhood absence epilepsy in the Chinese population may be linked to chromosome 8q24, the CAE gene is transmitted disequilibrium on locus D8S1783. Combined with other research results, we suppose that CAE gene may be in the ECA1 area on chromosome 8q24. (2) The CAE gene perhaps has a genetic heterogeneity in the population of different areas and different races. (3) HHRR and TDT seem to be the best statistical methods to do linkage disequilibrium study in the trios family.

Child ; Child, Preschool ; China ; Chromosomes, Human, Pair 8 ; genetics ; Epilepsy, Absence ; genetics ; Family Health ; Female ; Haplotypes ; genetics ; Humans ; Linkage Disequilibrium ; genetics ; Male ; Microsatellite Repeats ; Nuclear Family ; Risk Factors

Objective To study the effect of carbon monoxide (CO) on hydrogen sulfide (H2 S) in guinea pigs with allergic rhinitis (AR) through intervention treatment. Methods AR model in guinea pigs was established by using ovalbumin. The animals were divided into three groups. Group one was sensitized continuously by ovalbumin, group two was treated with Hemin as induction group, and group three was treated with zinc protoporphyrin (ZnPP) as suppression group. The guinea pigs treated with saline were used as control. The behavior science scores, eotaxin concentration of nasal lavage, IgE in blood serum were recorded, and the plasma concentrations of CO and H2S were determinated, then the expression of hemeoxygenase (HO) -1, cystathionine-β-synthase (CBS) and cystathionine-γ-lyase (CSE) were measured in nasal mucosa by fluorescent quantitative RT-PCR. Results The behavior science scores, concentration of eotaxin in nasal lavage, IgE in blood serum and concentration of CO in plasma of sensitized group were higher than those of control (P<0.01), and the expression of HO-1 in nasal mucosa was also higher than control [(7.61±2.80)×10-3 vs (2. 32±1.14)×10-3, p <0.05]. All these items were higher when treated with Hemin and lower when treated with ZnPP (P<0.05). The concentration of H2S in plasma was lower than control with significant differences [(14. 80±1.60)μmol/L vs (18. 90±1.00)μmol/L, P <0.01], the expression of CSE was also lower than control (P<0.05), and both of them were lower with Hemin induced and higher with ZnPP (P<0.05). The expression of CBS was very low and had no significant differences between groups (P>0.05), so it indicated that the CSE was the key enzyme for endogenous H2S product in nasal mucosa. Moreover the concentration of H2S was negatively correlated with CO (r = -0.702, P<0.001). Conclusions Endogenous CO and H2S play a significant role in the pathogenesis of AR, and HO-1 and CSE are the main speed-relate enzymes respectively . The H2S is also influenced by CO.

Objective To study the impact of carbon monoxide(CO)on expression levels of inducible nitric oxide synthase(iNOS)mRNA in guinea pigs with allergic rhinitis(AR).Methods Twenty four guinea pigs were divided randomly into four study groups with 6 guinea pigs in each.The guinea pigs in the first group were treated with saline only(Group 1,the healthy controls).The remaing guinea pigs were sensitized by ovalbumin and thus establishing the AR models.After sensitization,the animals in the second group remained untreated(Group 2,AR control group).The third group was treated with Hemin as the induction group,and the fourth group was treated with Zinc protoporphyrin(ZnPP)as the suppression group.The plasma concentration of carboxyhemoglobin(COHb)was measured,which represents the concentration of CO.The expression levels of Heme oxygenase-1(HO-1)and NOS mRNAs in nasal mucosa were determined by fluorescent quantitative RT-PCR.Results AR models were established successfully in all study guinea pigs.The concentrations of COHb(x-±s)in plasma of the second group(2.27% ±1.13%)were significantly(q=4.10,P＜0.01)higher than those of healthy controls(1.08% ± 0.24%).The plasma concentration of COHb in the third group(3.17% ±0.68%)were also significantly higher(q=3.12,P＜0.05)than those in the second group.The expression levels of HO-1 and iNOS in nasal mucosa of the second group[(7.80 ± 1.60)×10~(-3) and(5.81 ±0.05)×10~(-3),respectively]were also significantly(q equals 5.52 and 7.21,respectively,P＜0.01)higher than those of controls[(1.96 ±0.71)×10~(-3) and(0.97 ±]0.05)×10~(-3),respectively].The expression levels of HO-1 and iNOS in the nasal mucosa of the third group[(11.89 ± 4.78)×10~(-3) and(7.42 ± 0.70)×10~(-3),respectively]were significantly(q equals 3.86 and 2.22,P＜0.05)higher than those of the second group.The expression levels of HO-1 and iNOS in nasal mucosa of the fourth group[(3.82 ±0.98)×10~(-3) and(2.34 ±0.04)×10~(-3),respectively]were significantly(q equals 3.76 and 5.18,P＜0.05)lower than those in the second group.Conclusions Endogenous carbon monoxide influenced the expression levels of iNOS in nasal mocusa in guinea pigs with AR.