Objective To investigate the effect of cigarette smoke exposure on the interleukin-17-producing CD4+T helper lymphocyte( Th17 ) in peripheral blood and lungs of cigarette smoke-induced emphysema mice. MethodsForty male BALB/c mice were randomly divided into 4 groups, including a control group C12, a control group C24, a smoke-exposure group S12 and a smoke-exposure group S24, 10 mice each group. Morphological changes were evaluated by mean linear intercepts (Lm) and destructive index (DI). The concentration of IL-17, IFN-γ and TNF-α in bronchoalveolar lavage fluid(BALF) and the concentration of IFN-γ and TNF-α in lung homogenates were measured by enzyme-linked immunosorbent assay (ELISA). The proportion of CD4+IL-17+ Th17 cells in peripheral blood and lungs of mice was determined by flow cytometry. The mRNA expressions of RORγt and IL-17 in peripheral blood and lungs were evaluated by real-time PCR. ResultsLm and DI were significantly higher in S12 and S24 groups [(39.19±3.51)μm vs (46.87±7.16) μm, 39.13±1.57 vs 45.16±3.13]than that in C12 and C24 groups [(32.60± 3.21 ) μm vs (32.38±3.73) μm, 28.23± 1.62 vs 28.86±2.07], all P＜0.05. IL-17, IFN-γand TNF-α in BALF of S12 and S24 groups [(119.72±10.72) ng/L vs (296.40±14.00) ng/L,(129.7±22.2) ng/L vs(251. 1±62.4) ng/L, (17.35±1.60) ng/L, (36.35±1.43) ng/L]were higher than those in C12 and C24 groups [(52.06±4.70) ng/L vs (51.89 ±6.82) ng/L, ( 85.8 ±26.8) ng/L vs ( 88.9 ± 11.5 ) ng/L,(6.41 ±0.90) ng/L vs (5.85±0.92) ng/L], IFN-γ and TNF-α in lungs of S12 and S24 groups [( 1124.3±174.4) ng/L vs (1342.7±206.1) ng/L, (77.2±13.7) ng/L vs (101.7±19.0) ng/L, (129.7±22.2)ng/L vs (251.1 ±62.4) ng/L]were higher than those in C12 and C24 groups[(946.2±81.9) ng/L vs (1027.2±188.3) ng/L, (62.1±16.1) ng/L vs(64.4±15. 1) ng/L], all P＜0.05. The percentage of Th17 cells in lungs of S12 and S24 groups [(3.27±1.12)％ vs (7.19±2.24)％, ( 1.96±0.61 )％ vs (3.82±1.26)％]was significantly increased as compared with that of C12 and C24 groups [(1.80± 0.75)％ vs (1.99±0.59)％], all P＜0.05. And the percentage of Th17 cells in peripheral blood of S12 and S24 groups [(1.96±0.61)％ vs ( 3.82± 1.26)％]was also significantly higher than those in C12 and C24 groups [(0.90±0.37)％ vs (0.97±0.32)％], all P＜0.05. In peripheral blood and lungs,the mRNA levels of RORγt and IL-17 in S12 and S24 groups were higher than in C12 and C24 groups, all P＜0.05.Moreover, the frequency of Th17 cells in peripheral blood and lungs had a positive correlation with Lm and DI( r=0.706-0.772 ,all P＜0.05 ). ConclusionAn up-regulation of proportions Th17 in peripheral blood and lungs of cigarette smoke-inducsed emphysema mice were detected. It may be involved in the amplified and persistent inflammation.

Objective To evaluate the expression of Tc17 in a cigarette smoke-induced mice model of emphysema.To explore the probable mechanisms about how Tc17 cells to elevate in lungs of mice.Methods Forty male Balb/c mice were randomly divided into four groups,including control group ( 12 weeks,C12),control group (24 weeks,C24),smoke-exposure group (12 weeks,S12) and smoke-exposure group (24 weeks,S24 ),10 mice each group,Emphysema of mice was observed by HE pigmentation.Morphological changes were evaluated by mean linear intercepts (Lm) and destructive index (DI).The proportion of CD8+ IL-17 + Tc17,CD8+ IL-17 + CC chemokine receptor type 6 ( CCR6 ) + and 6CCR6 + Tc17 cells in lungs of mice was determined by flow cytometry.The mRNA expressions of retinoidrelated orphan nuclear receptor(RORγt) and IL-17 were evaluated by real-time PCR.The levels of IL-1 β,IL-6,IL-23,transforming growth factor β (TGFβ) and CC chemokine ligand 20 (CCL20) were tested by ELISA.Correlations among these indexes were analyzed.Results Lm and DI were significantly higher in S12 and S24 than in C12 and C24,S24 in particular (t value 4.378-15.188,all P ＜ 0.05).The percentages of Tc17 in S12 and S24[(9.28 ± 1.12)%,( 13.13 ±3.56)%]was significantly increased as compared with that in C12 and C24[(2.40 ±0.60 )%,(2.64 ±0.96 )%],S24 in particular.The mRNA levels of RORγt and IL-17 in S12 and S24 were higher than in C12 and C24,S12 and S24 in particular.There was significant difference (all P ＜0.05 ).The frequency of Tc17 cells had a positive correlation with Lm and DI ( r value were 0.734 and 0.884 respectively,P ＜ 0.01 ).The percentages of CD8+ IL-17 + CCR6 +T cells and CCR6 + Tc17 were significantly elevated in S12 and S24 compared to C12 and C24,S24 in particular (all P ＜ 0.05 ).There was positive correlation between Tc17 cell ratio and CCL20 levels( r =0.899,P ＜0.01 ).The levels of IL-1 β,IL-6,IL-23 and TGFβ in S12 and S24 were significantly increased as compared with that in C12 and C24.There was significant difference (all P ＜0.05).Meanwhile,the frequency of Tc17 cells had a positive correlation with IL-1β,IL-6,IL-23,and TGFβ.Conclusions An up-regulation of proportions Tc17 in lungs of cigarette smoke-induced emphysema mice were detected.The CCR6/CCL20 axis and the increased IL-1β,IL-6,IL-23 and TGFβ probably contributed to this up-regulation.

Objective To investigate the relationsheep between colonic H2 S‐producing enzyme cystathionine‐γ‐lyase(CSE) and colonic motility in a rat model of diabetes mellitus(DM) .Methods To obtained diabetic rat models ,all rats were injected intra‐peritoneally (ip) 1% streptozotocin (STZ ,65 mg/kg) ,type 1 diabetes model was established ,nornmol conditions were observed pe‐riodically .Ten days after treatment ,production of colonic longitudinal smooth muscle strips ,organ bath recordings were used to test the colonic motility ;immunofluorescence and double immunofluorescence lableling method were performed to detect the distribution of CSE;Western blot were performed on rat colonic samples devoid of mucosa and submucosa to detect the expression of CSE .Re‐sults DM rats decreased the colonic motility(P<0 .05);CSE was strongly expressed in the cytosols of the circular and longitudinal smooth muscle cells and the nucleus of the myenteric plexus neurons ;CSE distributed in myenteric neurons in the nucleus of rat ,but the proportion of MP had no difference (P>0 .05);DM rats increased the expression of CSE in the colon devoid of mucosa and sub‐mucosa(P<0 .05) .Conclusion The decrease of colonic motility in diabetic rat may be associated with the increased production of H2 S‐producing enzyme CSE .

In industrial glutamate fermentation, intermitted feeding glucose with the help of off-line glucose measurement is generally necessary. This kind of feeding strategy could cause large variations in glucose concentration so that it is not favorable for the achievement of efficient and stable glutamate fermentation. Glutamate fermentation is characterized with typical non-growth association behavior, and during glutamate production phase glucose consumption is closely correlated with ammonia consumption. In this study, glucose concentration was controlled at various pre-determined levels by predicting glucose consumption amount and thus its concentration with the aid of on-line monitoring ammonia consumption. When glucose concentration was controlled around a lower level of 5 g/L~10 g/L, the final glutamate concentration could reach a relatively higher level of 80 g/L. In this way, the huge osmotic stress change due to the large glucose concentration variation with the intermitted feeding method could be avoided and the glutamate fermentation performance enhancement be expected.

OBJECTIVETo explore clinical efficacy of Yiguanjian Decoction (YD) combined Adefovir Dipivoxil Tablet (ADT) in treating HBeAg negative chronic viral hepatitis B (CVHB) active compensated liver cirrhosis (LC) patients.

METHODSTotally 68 HBeAg negative CVHB active compensated LC patients initially treated were assigned to the treatment group and the control group using random digit table, 34 in each group. Patients in the control group took ADT alone, 10 mg each time, once per day. Those in the treatment group additionally took YD, one dose per day. The therapeutic course for all was 48 weeks. Levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil) were detected once in every two weeks. Hepatitis B virus (HBV)-DNA and four items of serum liver fibrosis [procollagen type I (PCN), hyaluronidase (HA), procollagen III peptide (PCIII), laminin (LN)] were detected once per every 4 weeks. Abdominal ultrasound B was performed before and after treatment. The inner diameter of the portal vein and the size of spleen were recorded. The fibrosis degree of liver was evaluated using Fibroscan. Efficacy of Chinese medicine (CM) was evaluated between the two groups before and after treatment using CM syndrome integrals. Efficacy of Western medicine (WM) was also evaluated between the two groups using Child-Pugh grading. Results Compared with before treatment in the same group, ALT and AST levels restored to normal levels, HBV-DNA turned negative (HBV-DNA < or = 1 x 10(2)) in the two groups after 48-week treatment. Besides, levels of TBil, ALB, PCIV, HA, PCIII, and LN obviously decreased (P < 0.05, P < 0.01). Results of ultrasound B showed the inner diameter of the portal vein and the size of spleen decreased. Fibroscan results showed that the elasticity value of the liver obviously decreased (P < 0.05). Besides, post-treatment levels of PCIV, HA, PCEJ, and LN, and the elasticity value of the liver decreased more obviously in the treatment group than in the control group (P < 0.01). There was no statistical difference in post-treatment levels of ALT, AST, TBil, ALB, inner diameter of the portal vein, or the size of spleen between the two groups (P > 0.05). Compared with before treatment in the same group, scores of Chinese medical syndrome and Child-Pugh scores decreased in the two groups after treatment (P < 0.05, P < 0.01). Besides, scores of Chinese medical syndrome decreased more obviously in the treatment group than in the control group (P < 0.05). The effective rate was 8824% (30/34) in the treatment group, higher than that of the control group [67.65% (23/34)] with statistical difference (P <0.05). Conclusion Combined treatment of YD and ADT could significantly improve symptoms of CM and fibrosis degree of liver of HBeAg negative CVHB active compensated LC patients.

Adenine ; analogs & derivatives ; therapeutic use ; Alanine Transaminase ; blood ; Antiviral Agents ; therapeutic use ; Aspartate Aminotransferases ; blood ; Bilirubin ; blood ; DNA, Viral ; blood ; Drugs, Chinese Herbal ; therapeutic use ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; drug therapy ; Humans ; Liver Cirrhosis ; drug therapy ; virology ; Organophosphonates ; therapeutic use ; Tablets

OBJECTIVETo explore the technique of induction of polyploidy in Salvia bowleyana by colchicine treatment.

METHODThe three kinds of explant of bud, leaf and calli were induced by colchicine treatment.

RESULTThe induction effects were better when the calli was treated by colchicines (15 mg x L(-1)) and the leaf was pre-cultured for one week. The doubling rate was 33.33%, while the majority were wholy doubled plants, and the leaves were thicker and broader, the color was darker, the root was thicker and the stoma size was obviously bigger than the diploid plants. The number of chromosome were 8 to 64. Isoenzyme analysis showed that the enzyme activities between the polyploid and the diploid plants were quite different.

CONCLUSIONInduction of polyploidy by colchicine treatment is efficacious. The part of the doubled plants were identified as homologmous tetraploids.

Chromosomes, Plant ; genetics ; Colchicine ; pharmacology ; Plant Leaves ; anatomy & histology ; genetics ; growth & development ; Plant Shoots ; anatomy & histology ; genetics ; growth & development ; Plants, Medicinal ; anatomy & histology ; genetics ; growth & development ; Polyploidy ; Salvia ; anatomy & histology ; genetics ; growth & development

AIMTo identify the main metabolites of aconitine in the urine of rabbits.

METHODSAfter oral administration of aconitine (5 mg.kg-1), the urine of male rabbits was collected and extracted by solid phase extraction and analyzed by liquid chromatography-ion trap mass spectrometry.

RESULTSAconitine and 4 metabolites were found in the rabbit urine. Their protonated molecular ions at m/z 632, m/z 604, m/z 590, m/z 500 and multistage fragment ions with neutral loss of 60 u, 32 u, 28 u and 18 u were monitored. Their relative concentration were M1 > Aconitine > M4 > M2 > M3.

CONCLUSIONThe metabolites M1-M4 were deduced as 16-O-demethylaconitine, benzoylaconine, 16-O-demethylbenzoylaconine and aconine, respectively.

Aconitine ; analogs & derivatives ; metabolism ; urine ; Alkaloids ; urine ; Animals ; Chromatography, High Pressure Liquid ; Male ; Rabbits ; Spectrometry, Mass, Electrospray Ionization

Glutamate dehydrogenase (GDH) is a key enzyme in the biosynthesis of glutamate. The GDHs from Corynebacterium glutamicum S9114 the most commonly used strain in glutamate fermentation, were purified and their molecular structures and properties characterized. The coenzymes were also studied in the hope to increase glutamate production. Cells were harvested at mid-exponential phase by centrifugation and washed with Tris-HCl buffer containing DTT and EDTA (pH 7.5). The cells were then disrupted using a French pressure cell press and the supernatant was collected by centrifugation. The extract was concentrated by 70-fold using the AKTA-100 FPLC system employing a DEAE-cellulose ion exchange column, a hydrophobic interaction chromatography (HIC) and Sephadex G-200 gel filtration. The purified extracts contained NADPH-dependent GDH and NADH-dependent GDH. Both of the enzymes were highly specific for the coenzymes. The molecular masses of the NADPH-dependent GDH and its subunit were 188kD and 32kD respectively, suggesting the enzyme is a homo-hexamer. Our data reported for the first time the presence of NADH- dependent GDH in Corynebacterium glutamicum S9114, similar to other microorganisms containing both GDHs. The NADPH-dependent and NADH-dependent GDH in Corynebacterium glutamicum S9114 may participate in the assimilation and dissimilation of ammonia respectively. The absorptions of NADPH-dependent GDH was very weak at 280nm but very high at 215nm, suggesting a low phenylalanine and tyrosine content in the enzyme.
Chromatography, Gel ; Chromatography, Ion Exchange ; Corynebacterium glutamicum ; enzymology ; Glutamate Dehydrogenase ; isolation & purification ; metabolism ; Molecular Weight ; NADP ; metabolism ; Substrate Specificity

Haizao Yuhu decoction (HYD) is a formula that has been used for approximately 500 years and famous for its efficiency in treating thyroid-related diseases in traditional Chinese medicine (TCM). HYD was first presented by Chen Shi-gong in a famous surgical monograph named Waike Zhengzong during the Ming Dynasty. We conducted the research to investigate the possible pharmacokinetic profile of different prescriptions of HYD in rats, in order to reveal the interactions of Haizao and Gancao drug pair with other herbs in HYD. Liquiritin, naringin, besperidin, peimine, peiminine liquiritigenin, glycyrrhizic acid, hergapten, nobiletin, osthole, glycyrrhetinic acid in blood samples were determined by UPLC-MS/MS. The result revealed tbat Haizao could enhance the peak concentration of glycyrrhizic acid. The other herbs in HYD may promote'the absorption of flavonoids in Gancao in normal rats, but inhibit the absorption of saponins and accelerate their metabolism. Gancao and Haizao drug pair could enhance the bioavailability of hesperidin, peimine, bergapten, nobiletin and osthole and prolong the elimination of peimine and naringin.
Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Magnetic Resonance Spectroscopy ; Male ; Mass Spectrometry ; Plasma ; chemistry ; Rats ; Rats, Wistar

DNA, Ribosomal ; genetics ; Genome, Plant ; Phylogeny ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide ; genetics ; RNA, Ribosomal, 5S ; genetics ; Sequence Homology, Nucleic Acid ; Species Specificity ; Valerian ; classification ; genetics