Objective To study the constituents in Melicope pteleifolia. Methods Plant material was isolated with 80% EtOH. Compounds were separated with chromatographic methods and their structures were elucidated on the basis of spectral analysis (EI-MS, 1H-NMR, and 13C-NMR) and chemical evidence. Results Five compounds were isolated from petrol ether or ethyl acetate soluble fraction. Their structures were identified as 3,5,3'-trihydroxy-4'-methoxy-7-(3-methylbut-2-enyloxy) flavone (pteleifolosin C, 1), 3,7-dimethoxyl kaempferol (kamatakenin, 2), vanillic acid (3), tricosanoic acid tetradecyl ester (4), and p-sitosterol (5), respectively. Conclusion Compound 1 is a new structure named pteleifolosin C. Compounds 2-4 are isolated from this plant for the first time.

Objective To investigate the application value of perioperative enhanced recovery after surgery (ERAS) combined with laparoscopic common bile duct exploration (LCBDE) in the treatment of choledocholithiasis.Methods The clinical data of 84 patients with choledocholithiasis who were admitted to the Yijishan Hospital from January 2011 to December 2013 were prospectively analyzed.A single-blind,randomized,controlled study was performed in the 75 patients who were allocated into the control group and the enhanced recovery after surgery group (ERAS group) based on a random number table.All the patients underwent LCBDE,the patients in the control group received conventional perioperative management and the patients in the ERAS group received perioperative management according to enhanced recovery rehabilitation program.All the patients were followed up by outpatient interview till postoperative month 6.The clinical features,liver function and residual stones in the patients were observed.The operation time,postoperative complications,postoperative intestinal function recovery,duration of hospital stay and hospital expenses in the two groups were compared.Measurement data with normal distribution were presented as x ± s.Comparison between groups were evaluated with an independant sample t test.Count data were analyzed using the chi-square test.Results All the 75 eligible patients undergoing successful operation were randomly divided into the control group (35 patients) and the ERAS group (40 patients).The operation time and volume of intraoperative blood loss in the control group and the ERAS group were (185 ±46)minutes and (124 ±28)mL,(178 ±37) minutes and (114 ±32)mL,respectively,with no significant difference (t =0.729,1.431,P ＞ 0.05).There were 12,14 and 10 patients in the control group and 5,6 and 4 patients in the ERAS group with postoperative incision pain,vomit and infection,showing a significant difference (x2=5.054,5.966,4.241,P ＜ 0.05).The level of white blood cell,alanine aminotrausferase and direct bilirubin in the control group and in the ERAS group were (11.4 ± 3.5) × 109/L,(128 ± 33)U/L,(38 ±14) μmol/L and (10.6 ± 3.0) × 109/L,(135 ± 35) U/L,(44 ± 16) μmol/L at postoperative day 1,compared with (7.8 ±2.9) × 109/L,(48 ± 14) U/L,(21 ± 8) μmol/L and (6.9 ±2.1) × 109/L,(43 ± 13) U/L,(20 ±7) μmol/L in the 2 groups at postoperative day 4,respectively,showing no significant difference between the 2 groups (t =1.018,-0.872,-1.767,1.553,1.836,1.044,P ＞ 0.05).The postoperative first flatus day,time of food intake,time of postoperative infusion and duration of hospital stay were (42 ± 13)hour,(45 ±14) hours,(6.8 ±2.3)days and (11.3 ±4.5)days in the control group,and (35± 11)hours,(19 ±7)hours,(4.2 ± 1.8) days and (9.6 ± 2.4) days in the ERAS group,with a significant difference between the 2 groups (t =2.741,10.524,5.485,2.077,P ＜ 0.05).The total hospital expenses was (18 729 ± 3 127) yuan in the control group,which was significantly greater than (16 981 ±2 756) yuan in the ERAS group (t =2.574,P ＜ 0.05).The liver function of all the patients was recovered at the postoperative month 1.Four patients with residual stones in the 2 groups were detected by T-tube cholangiography,and were cured by removal of gallstones by choledochoscopy.There were no complications of the abdominal pain,jaundice and fever in all the patients till the end of follow-up.Conclusion ERAS combined with LCBDE for the treatment of choledocholithiasis is safe and feasible,with the advantages of low morbidity,quick recovery,short duration of hospital stay and less hospital expenses.

Objective To study the apoptosis of multiple myeloma cell line KM-3 induced by NK cells. Methods WST-1 assay was used to detect the killing effect of KM-3 cells treated with NK cells at different effector(E):target(T) ratio. Flow cytometry was applied to analyze Annexin-V+/PI- apoptotic cells and the mitochondrial transmembrane potential. Results NK cells could significantly kill KM-3 cells in a dosand time-dependent manner (P <0.05). After KM-3 cells- were treated with NK cells for 48 hours, the Annexin-V+/PI- cells were increased obviously in dose-dependence (P <0.05). The Annexin-V+PI- cells were increased in time-dependence when treated with NK cells(E:T ratio at 10:1) (P<0.05). The mitochondrial transmembrane potential of KM-3 cells treated with NK cells were significantly decreased in dose-and time-dependence (P < 0.05). Conclusion NK cell can kill KM-3 cells and induce apoptosis in a dose-and time-dependence manner.

Objective To study the chemical constituents of Evodia lepta.Methods The chemical constituents were isolated by chromatographic methods and their structures were elucidated by physicochemical characteristics and spectral data.Results Two compounds were isolated and their structures were identified as erythro-3-(1',2',3'-trihydroxy) isopentyl-7-hydroxycoumarin(Ⅰ) and?-daucosterol (Ⅱ).Conclusion CompoundⅠis a new one named evodosin A while compoundⅡis isolated from E. lepta for the first time.

Objective To investigate epidemiological and clinical features of human ehrlichiosis.Methods The epidemiological, clinical, laboratory, therapeutic and prognostic data of 21 clinically diagnosed cases of human ehrlichiosis were retrospectively analyzed. Results The epidemic regions where the ticks' activity was high located at the boundary between Hubei and Henan Provinces. All cases were farmers. The median age was 50 years ranged from 19 to 69 years. The male female ratio evident history of tick bite 1 week before the onset. The common symptoms included fever, diarrhea,cough, nausea and vomiting, abdominal pain and expectoration. The complications included hemorrhage, toxic encephalopathy, acute renal insufficiency, secondary infection and respiratory failure. The common abnormalities of routine lab data were thrombocytopenia, hypoeosinophilia,elevated lactate dehydrogenase, creatine kinase and aminotransferases, leucopenia and proteinuria.Nine cases were tested with peripheral blood smear and intracytoplasmic inclusions in neutrophils were found in one case. Seventeen cases were tested with serological assay and antibodies against Ehrlichia were positive in five cases. After doxycycline, symptomatic and supportive treatments, 14 cases were recovered and seven died. The average age of the deaths was 56 years. Conclusions Human ehrlichiosis is an acute tick-borne zoonosis and multiorgan could be involved. The older cases prone to develop complications and the prognosis is poor.

Objective To study the relation between alcohol-induced osteonecrosis of femoral head (AIONFH) related with high morbidity TCM constitution type with CYP2C8 gene polymorphisms.Methods Totally 152 Han nationality NONFH cases from Feburary 2014 to September 2015 from outpatient and the inpatient departments in Gansu Province Hospital of TCM were collected. 50 AIONFH cases were set as medical case group; meanwhile, 45 healthy volunteers were enrolled as control group. Database for medical materials of all patients and volunteers was established. TCM distribution for AIONFH patients was determined. Solution DNA extraction kit was used to extract DNA, and detect the concentration and purity of DNA. The target gene was amplified by PCR and the target gene was amplified by gel electrophoresis. The length of the fragment was confirmed to conduct target gene sequencing. With the results of sequencing and gel electrophoresis, the relation of AIONFH with CYP2C8 gene polymorphism in AIONFH patients with phlegm-dampness syndrome and the control group.ResultsThe CYP2C8 gene loci rs17110453 gene polymorphism was not statistically significant between the two groups (χ2=0.253,P>0.05). There was no significant difference in allele between the two groups (χ2=0.077,P>0.05). The risk of disease in CC genotype was 1.37 times higher than the AA genotype (95%CI: 0.339-5.540), without statistical significance (P>0.05). There was no significant difference in genotype and allele distribution between AIONFH patients with phlegm-dampness and non-phlegm-dampness and the control group (P>0.05).Conclusion CYP2C8 gene loci rs17110453 gene polymorphism A/C mutation has no obvious relation with AIONFH risk. There is no clear relationship between CYP2C8 gene loci rs17110453 gene polymorphism with AIONFH.

Objective To identify the important regulatory elements in the promoter of human CD2 associated protein(CD2AP) by conserved sequence analysis among different species and luciferase functional detection. Methods The promoter sequences of CD2AP from different species were analyzed by BLAST. Plasmids containing different length of deletion mutations of human CD2AP promoter were constructed. Pro-moter activities were tested in 3 kinds of cells from different species by luciferase analysis and were tested in HEK-293 cells treated with all-trans-retinoic acid. Results Homologous sequence comparison in CD2AP promoter area among human, cattle and pig showed that putative specific protein 1 (Sp1) sites and down-stream promoter element (DPE) were highly evolutional]y conserved. Progressive deletion luciferase analysis of DNA fragments revealed similar promoter activity style among 3 different cell lines from 3 different spe-cies, HEK-293, BHK-21 and Vero cells. One basic promoter activity located within 500 bp upstream of ATG. Fragments of further upstream 100 bp or more had drastically 10 times increased promoter activity. Two putative Sp1 sites were in this 100 bp region. All-trans-retinoic acid decreased the luciferase activity of CD2AP promoter. Conclusion Putative Spl sites and DPE have important functions in the promoter activity of CD2AP.

A method for quantitation of collagen was established by detecting marker peptide with high performance liquid chromatography-mass spectrometry (HPLC-MS). Theoretical marker peptides were selected by sequence comparison. Bovine collagen type I was digested with trypsin. Marker peptides typical for collagen type I were identified with HPLC-MS. The relationship between the abundance of marker peptides and collagen concentration was established. The results show that GEAGPSGPAGPTGAR and the other 5 peptides showed high resolution during chromatographic separation and high signal intensity during MS analysis. Peptide signal intensity and collagen concentration showed a good linear relationship in the range from 0.1 to 3 mg/mL. Bovine tendon and collagen sponge were used as actual samples and collagen contents were determined as 90.2% and 93.4% respectively. Quantitation of marker peptides of collagen was a feasible method to identify and quantify collagens in medical device research and development.
Animals ; Cattle ; Chromatography, High Pressure Liquid ; Collagen Type I ; analysis ; Mass Spectrometry ; Peptides ; analysis

OBJECTIVETo investigate the relationship between the variation of chloroplast DNA gene sequences and the geographical origins of Polygonum capitatum in order to provide the molecular evidence for its excellent germplasm resources.

METHODPCR direct sequencing was applied to detect the chloroplast psbA-trnH, trnL-trnF gene sequence of 11 samples collected from 11 populations of P. capitatum.

RESULTThe psbA-trnH gene sequence of P. capitatum from different populations was 402 bp in length, there were 6 variable sites. TrnL-F gene sequence was 875 bp, there were 5 variable sites. The clusters diagram by UPGMA method showed that P. capitatum groups in Yunnan and Guizhou existed a considerable variation.

CONCLUSIONP. capitaturni which is located in the east of Yunnan and the west of Guizhou is helpful of screening the germplasm resources.

Alleles ; Base Sequence ; DNA, Chloroplast ; genetics ; Molecular Sequence Data ; Mutation ; Phylogeny ; Polygonum ; classification ; genetics ; Sequence Alignment ; Sequence Analysis, DNA

OBJECTIVETo detect genetic diversity of 48 population of Polygonum capitatum in Guizhou province.

METHODThe genetic diversity of 48 representational populations of P. capitatum including 240 individuals had been investigated by ISSR marker technique.

RESULTThe genetic diversity had been revealed as follow: A total of 8 293 bands were produced in 240 individuals, of which 7 962 bands were common in the 48 population. The value of the average percentage of polymorphic bands (PPB) was 79.09%, Nei's genetic diversity index (H(e)) was 0.245 8, Shannon's information index (I) was 0.396 2, and genetic differentiation index (G(st)) was 0.238 0 at population level, respectively. The genetic differentiation index (G(st)) was 0.072 2, genetic differentiation coefficient by Shannon's diversity (I(st)) was 0.044 2 within the population levels. Groups cluster analysis based on the UPGMA method indicated that although the 48 populations could be divided into 3 groups and the P. capitatum seed sources. The groups cluster showed that a cross clustering of P. capitatum between the southwest and southeast populations in Guizhou province, and no significant correlation was found between geographical and genetic distance among them.

CONCLUSIONThe genetic diversity of P. capitatum is relatively high at the population levels, while low within the population levels, a significant degree of genetic differentiation occurs among the populations. The groups cluster analysis indicated they has not apparent genetic variation in regional pattern between the place of origin populations and the migrate populations.

China ; Genetic Variation ; Molecular Sequence Data ; Phylogeny ; Polygonum ; classification ; genetics ; Repetitive Sequences, Nucleic Acid