ObjectiveTo establish an accurate method for simultaneous determination of plasma Kyn and Trp by HPLC-UV detection.Methods Kyn and Trp were separated on Agilent Hypersil ODS column using 3-nitrotyrosine as internal standard.The mobile phase consisted of 15 mmol/L sodium acetateacetic acid (pH 5.5):acetonitrile 94∶ 6(v/v) at a rate of 0.8 ml/min.The chromatographic separation was performed at 25 ℃.The eluate was monitored with programmed wavelength setting at 360 nm from 0 to 4 min for Kyn and at 302 nm from 4 to 5 min for Trp.The method was applied to determination of plasma Kyn and Trp in 8 chronic glomerulonephritis,10 idiopathic thrombocytopenic purpura,15 chronic hepatitis B virus patients and 15 healthy controls from September to December in 2010.The differences were compared using ANOVA and SNK methods.Results The retention time of Kyn and Trp were 2.9 min and 4.4 min,respectively.For Kyn,the assay was linear from 0.44 μmol/L to 18.30 μmol/L.For Trp,the linearity was from 3.67 μmol/L to 470.00 μmol/L.The detection limits were 0.014 μmol/L for Kyn and 0.122 μmol/L for Trp,respectively.The within-day CVs were ＜ 3％ and the between-day CVs were ＜ 4％.The mean recoveries yield were in the range of 92.29 to 104.40.The plasma concentrations of Kyn were ( 1.59 ± 0.28),(2.73 ± 0.56),(2.69 ± 0.44) and ( 1.54 ± 0.48 ) μmol/L,the plasma concentrations of Trp were (59.8 ± 10.0),(46.1 ± 11.7),(58.5 ±8.0) and (41.4±13.1) μmol/L,the Kyn/Trp were (0.027 4±0.007 5),(0.061 6 ±0.016 5),(0.046 7 ±0.009 1) and (0.038 3 ±0.007 5)in controls,chronic glomerulonephritis patients,idiopathic thrombocytopenic purpura patients and chronic hepatitis B virus patients,respectively.There were significance difference of Kyn,Trp and Kyn/Trp amony the four groups (F=23.734,8.463,20.921,all P＜0.01).Conclusion The method is simple,fast,and suitable for applicability to clinical measurement.

Background The main mechanism of diabetic retinopathy (DR) is the formation of retinal neovascularization. Vascular endothelial growth factors (VEGF) is primarily factor for DR. Objective The goal of this study was to investigate the effects of angelicae sinensis decoction for supplementing blood ( ASD), a kind of traditional prescription of Chinese herbal medicine, on vascular endothelial growth factor (VEGF) level in vitreous and serum of STZ-induced type 2 diabetes mellitus. Methods Thirty-four SPF male SD rats were fed using high fat diet for 8 weeks and then the streptozocin of 25 mg/kg was intraperitoneally injected to create the type 2 diabetic models. ASD (3.6 g/kg) and 10 mg/kg Gli of 5 mi were administered intragastrically from the 4th day of high fat die though 8 weeks respectively. Other 10 matched rats were used as normal control group. The body weight, blood glucose level ,triglycerides (TG) and high density lipoprotein (HDL) were detected in 4 and 8 weeks after usage of drug. The vitreous and serum samples were obtained in the 8 th weeks for the VEGF detection by enzyme-linked immunosorbent assay (ELISA). This experiment comlied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. Results The body weight and HDL in DM group were significantly lower than those of normal control group in 4 or 8 weeks ( P＜0. 01 ), but in ASD and Gli groups ,the body weight and HDL were higher than those of the DM group (P＜0. 05 ). the blood glucose level and TG were elevated in DM group compared with normal control group (P＜0. 01 ), but those in ASD group and Gli group were decreased in comparison with DM group ( P＜0.05 ). EIISA revealed that no significant differences were found in VEGF levels between normal control group and ASD group or Gli group (P＞0. 05 ), but in DM group, the serum VEGF level (147.65±4. 55 ng/L) was reduced and vitreous VEGF level (673.45± 100. 85 ng/L) was raised,showing the considerable differences (P＜ 0.05 ). No any correlation was found between serum VEGF level and vitreous VEGF level (r=0.314,P＞0. 05). Conclusion ASD can decrease the VEGF level in vitreous,implying that ASD postpone the occurrence of diabetic retinopathy in DM rat.

Objective To observe the expression and localiz at ion of tumor necrosis factor alpha (TNF-?) of granulation tissue and chole steatoma in chronic otitis media, and explore its relationship with chronic otit is media. Methods The expression of TNF-? was observe d by immunohistochemical method in cholesteatoma, granulation tissue and normal external ear canal skin in 42 cases with chronic otitis media. Results The level of TNF-? in the group of the sample s including cholesteatoma and granulation tissue increased significantly (P

Objective To evaluate the internal reliability of Psychosocial Function Evaluation for mental disabled people in day care unit. Methods 16 participants were evaluated twice by the same rater (social worker) in the unit at the Permanent stage and the Steering stage. Results Cronbach's α was greater than 0.7 in the both evaluation. The Spearman correlation coefficient between each dimension and total score were between 0.502 and 0.869. Conclusion The internal reliability of Psychosocial Function Evaluation is satisfactory for mental disabilities.

Objective To examine the changes of red blood cell levels in the obese and non-obese patients with coronary heart disease (CHD) and its clinical significance. Methods 230 cases of coronary heart disease were selected and divided into the obese group and the nonobese group. Obesity and non-obesity were defined based on the body mass index (BMI I (Y) 28.0kg/m2), or waist-hip ratio (men> 0.9, women> 0.85). In addition, 130 healthy subjects were recruited as controls. The pathological status of coronary lesions was quantitatively analyzed according to the Coronary Vascular Image Segmentation Evaluation Criteria (American Heart Association 1984) and the Gensini scoring system. Results of the changes of both the hemoglobin levels and the red blood cell count in the obese group, the nonobese group with CHD and the control group were compared. Besides, Multivariant Logistic Regression Analysis was applied to assess the correlation between the red blood cells and the coronary artery disease. Results The red blood cell count and the level of hemoglobin in the obese group with CHD was higher than that in the non-obese group with CHD [(4.35 ± 0.55) and (4.13 ± 0.56) 109/L; (136.71 ± 15.87) and (129.96 ± 16.23) g/L, P < 0.05 in both]; the proportion of acute coronary syndrome in the obese group with CHD was higher in the obese group with CHD than that in the non-obese group with CHD (P<0.05); Multivariant logistic regression analysis also showed that the red blood cell count was positively correlated with obesity with CHD.Conclusion The red blood cell count and the level of hemoglobin in the obese group were higher than those in the non-obese group; the increase of red blood cell count and hemoglobin level is one of the independent risk factors for the obese patients with CHD.

Xiatianwu tablet is based on the theory of traditional Chinese medicine (TCM), combined with modern TCM pharmacology and selected 33 famous traditional Chinese crude drugs to compose. Its recipe helps cure rheumatism, relax tendons, promote blood circulation to relieve pain, et al. Although Xiatianwu tablets are widely applied to clinical remedy such as rheumatic arthritis, lumbar disc hernia, osteoarthritis and so on, there is no report about its application in fracture. This article is to observe the efficacy of compound Xiatianwu tablets in elderly patients with osteoporotic distal radius fractures and its impact on the wrist function and complications. 180 elderly patients with osteoporotic distal radius fractures, from January 2011 to June 2014, were divided into observation group and control group by the method of random number table, each group had 90 cases. The control group were gave Caltrate D after manipulative reduction and plaster immobilization, observation group were treated with compound Xiatianwu tablets in the basis of the control group. Efficacy, wrist function and complication rates were observed in two groups after treatment. The excellent and good rate was 95.56% in observation group better than 77.78% in control group, the difference was statistically significant (χ2 = 4.712, P < 0.05). The complication rate in observation group was significantly lower compared with the control group (P < 0.05). This study shows that compound Xiatianwu tablets can improve the efficacy in elderly patients with osteoporotic distal radius fractures, reduce the incidence of complications and relieve the pain of patients which plays a significant role in improving the quality of life.
Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Fractures, Bone ; drug therapy ; Humans ; Male ; Middle Aged ; Osteoarthritis ; drug therapy ; Radius Fractures ; drug therapy ; Tablets ; administration & dosage ; Treatment Outcome

OBJECTIVETo study the effect of exogenous Ca2+ on photosynthetic parameters of Pinellia ternate and accumulations of active components under high temperature stress.

METHODThe pigment contents of P. ternata leaves, photosynthesis parameters and chlorophyll fluorescence parameters of P. ternata leaves, the contents of guanosine, adenosine and polysaccharide in P. ternata tubers were measured based on different concentrations of exogenous Ca2+ in heat stress when the plant height of P. ternata was around 10 cm.

RESULTThe contents of total chlorophyll and ratio of chlorophyll a/b were relatively higher by spaying Ca2+. Compared with the control, spaying 6 mmol x L(-1) Ca2+ significantly enhanced the net photosynthetic rate (Pn), transpiration (Tr) and stomatal limitation (L8), but reduced intercellular CO2 concentration (C) in P. ternata leaves. With the increase of Ca2+ concentration, maximal PS II efficiency (Fv/Fm), actual photosynthetic efficiency (Yield) and photochemical quenching coefficient (qP) initially increased and then decreased, however, minimal fluorescence (Fo) and non-photochemical quenching coefficient (NPQ) went down first and then went up. The contents of guanosine and polysaccharide and dry weight of P. ternata tubers showed a tendency of increase after decrease, and the content of adenosine increased with the increase of Ca2+ concentration. The content of guanosine and polysaccharide in P. ternata tubers and its dry weight reached maximum when spaying 6 mmol x L(-1) Ca2+.

CONCLUSIONWith the treatment of calcium ion, the inhibition of photosynthesis and the damage of PS II system were relieved in heat stress, which increased the production of P. ternata tubers.

Breeding ; Calcium ; pharmacology ; Chlorophyll ; metabolism ; Dose-Response Relationship, Drug ; Heat-Shock Response ; drug effects ; Organ Size ; drug effects ; Photosynthesis ; drug effects ; Pinellia ; drug effects ; growth & development ; metabolism ; physiology ; Plant Leaves ; drug effects ; growth & development ; metabolism

Aim To investigate whether the effect of Aβ25-35 on Bcl-2 and Bax gene transcription through DNA methylation in SH-SY5Y cell.Methods Differ-ent concentrations of Aβ25-35 (0, 25, 50 μmol? L-1 ) were treated with SH-SY5Y cells for 48 h or 72 h in vitro.The optimal concentration and time of Aβ25-35 in-duced SH-SY5 Y apoptosis were determined by MTT method.Protein expression levels of Bcl-2 and Bax of Aβ25-35-treated groups were determined by Western blot.Real time PCR was used to detect the mRNA lev-els of DNA methyltransferase including DNMT 1 , DN-MT3a, DMT3b, MeCP2. Methylation specific PCR ( MSP) was used to analyze the effect of Aβ25-35 media-ted Bcl-2 and Bax gene promoter methylation .Results 25 μmol? L-1 Aβ25-35 was exposed to SH-SY5Y cells for 72 h, MTT assay showed that cell viability was (68.49 ±9.83 )%, which was significantly reduced compared with the control group ( P <0.05 ) , indica-ting AD cell apoptosis model was successfully estab-lished.Bcl-2 expression of Aβ25-35-treated group was significantly reduced compared with the control group , on the contrary , the expression of Bax was significantly increased .Real-time PCR results showed that com-pared with the control group , DNMT1, DNMT3a, DMT3b, MeCP2 mRNA levels of the Aβ25-35-treated groups had no significant difference ( P>0.05 ); MSP results showed that Bcl-2 and Bax unmethylated ampli-fication was positive , methylated amplification was neg-ative in control group , Bcl-2 and Bax unmethylated amplification was positive and methylated amplification was negative in Aβ25-35-treated group.Conclusion DNA methylation of Bcl-2 and Bax gene promoter are not affected during Aβ25-35 induced SH-SY5Y cell ap-optosis .

The zinc-regulated transporters (ZRT), iron-regulated transporter (IRT)-like protein (ZIP) plays an important role in the growth and development of plant. In this study, a full length cDNA of ZIP encoding gene, designed as DoZIP1 (GenBank accession KJ946203), was identified from Dendrobium officinale using RT-PCR and RACE. Bioinformatics analysis showed that DoZIP1 consisted of a 1,056 bp open reading frame (ORF) encoded a 351-aa protein with a molecular weight of 37.57 kDa and an isoelectric point (pI) of 6.09. The deduced DoZIP1 protein contained the conserved ZIP domain, and its secondary structure was composed of 50.71% alpha helix, 11.11% extended strand, 36.18% random coil, and beta turn 1.99%. DoZIP1 protein exhibited a signal peptide and eight transmembrane domains, presumably locating in cell membrane. The amino acid sequence had high homology with ZIP proteins from Arabidopsis, alfalfa and rice. A phylogenetic tree analysis demonstrated that DoZIP1 was closely related to AtZIP10 and OsZIP3, and they were clustered into one clade. Real time quantitative PCR analysis demonstrated that the transcription level of DoZIP1 in D. officinale roots was the highest (4.19 fold higher than that of stems), followed by that of leaves (1.12 fold). Molecular characters of DoZIP1 will be useful for further functional determination of the gene involving in the growth and development of D. officinale.
Amino Acid Sequence ; Cloning, Molecular ; Dendrobium ; chemistry ; classification ; genetics ; metabolism ; Gene Expression Regulation, Plant ; Iron ; metabolism ; Membrane Transport Proteins ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Plants ; chemistry ; classification ; genetics ; Sequence Alignment ; Zinc ; metabolism

Mitogen-activated protein kinases (MAPKs) are important signaling transduction components well conserved in eukaryotes and play essential roles in various physiological, developmental and hormonal responses in plant. In the present study, a MAPK gene, designated as DoMPK4 (GenBank accession No. JX297597), is identified from a rare endangered medicinal orchid species D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. The full length cDNA of DoMPK4 is 1 518 bp in length and encoded a 369 aa protein with a molecular weight of 42.42 kD and an isoelectric point of 5.55. DoMPK4 protein contained a serine/threonine protein kinase active site (158-170), a MAP kinase site (71-174), and eight conserved motifs. DoMPK4 had a transmembrane (214-232) but no signal peptide. Multiple sequence alignment showed that DoMPK4 shared high identities (74.9%-80.6%) with MAPK proteins from various plants. Phylogenetic analysis demonstrated that DoMPK4 belonged to group A of the MAPK evolutionary tree, and is closely related to monocots. Real time quantitative PCR (qPCR) analysis revealed that DoMPK4 is differentially expressed among the five organs including leaf, stem, root, seed, and protocorm-like body (PLB). The transcription level of DoMPK4 is the highest in the PLBs with 17.65 fold, followed by seeds, roots, and stems with 5.84, 2.28, and 1.64 fold, respectively. The progressive enhancement of DoMPK4 transcripts in the developing PLBs compared to that in the germinating seeds, suggests a role of DoMPK4 during the development of embryogenic PLBs formation in D. officinale.
Amino Acid Sequence ; DNA, Complementary ; genetics ; DNA, Plant ; genetics ; Dendrobium ; enzymology ; genetics ; Gene Expression Regulation, Plant ; Mitogen-Activated Protein Kinases ; genetics ; metabolism ; Phylogeny ; Plant Leaves ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; metabolism ; Plant Stems ; metabolism ; Seeds ; metabolism ; Sequence Alignment