OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.

METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.

RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.

CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.

Androgens ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Genetic Vectors ; Humans ; Male ; MicroRNAs ; physiology ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; metabolism ; Transfection

Animalcule Identification and Drug Susceptivity System is designed with virtual instrument technology. The paper introduces main functions and the special feature of the software design. It discusses how to accomplish database operations and serial communication with LabVIEW and achieve remote distribution of data with DataSocket tchnology.

OBJECTIVE:To optimize the ultrasonic extraction process of total alkaloids from mulberry leaves. METHODS:Based on the single factor experiment,response surface method was adopted to investigate the effects of ultrasonic extraction time, ultrasonic power,ethanol volume fraction,solid-liquid ratio and pH of solvent on the extraction rate of total alkaloids from mulber-ry leaves,then test data were analyzed by Design Expert 8.0.5 software,and the optimized process was confirmed and verified. RE-SULTS:The multiple correlation coefficient of established binomial equation fitting model was 0.969 9;the optimized condition for extracting alkaloid from mulberry leaf was ultrasonic extraction time of 48 min,ultrasonic extraction power of 800 W,ethanol vol-ume fraction of 70%,material-liquid ratio of 1∶25,pH 5. Under these conditions,the extraction rate of total alkaloids was 0.422%, with the bias ratio was less than 2% compared with the model predictions. CONCLUSIONS:The established model has good fit-ting performance,the extraction process is stable and reliable,and can be used for the extraction of alkaloids from mulberry leaves.

Chromogranin A ; metabolism ; Diagnosis, Differential ; Duodenal Neoplasms ; metabolism ; pathology ; surgery ; Ganglioneuroma ; metabolism ; pathology ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; Humans ; Male ; Middle Aged ; Neurofibroma ; metabolism ; pathology ; Paraganglioma ; metabolism ; pathology ; surgery ; Phosphopyruvate Hydratase ; metabolism ; S100 Proteins ; metabolism

OBJECTIVETo study the gene expression of the resistin and the effects of conjugated linoleic acid on its expression in white adipose tissue of obese rats fed with high fat diet during the formation of insulin resistance.

METHODSMale Wistar rats were randomly separated in control group, high-fat group and high fat + conjugated linoleic acid (CLA) group (0.75 g, 1.50 g, 3.00 g per 100 g diet weight), using reverse transcription polymerase chain reaction (RT-PCR) technique to measure the expression level of resistin and peroxisome proliferator-activated receptor-gamma (PPARgamma) mRNA expression.

RESULTSthe serum insulin and glucose levels of obese rats were (11.11 +/- 2.73) mIU/L, (5.09 +/- 0.66) mmol/L, and supplement of CLA might decrease hyperinsulinemia and hyperglycemia, in CLA group (0.75 g, 1.50 g, 3.00 g per 100 g diet weight) the serum insulin levels were (6.99 +/- 1.77) mIU/L, (7.36 +/- 1.48) mIU/L, (7.85 +/- 1.60) mIU/L, and glucose levels were (4.28 +/- 0.72) mmol/L, (4.18 +/- 0.55) mmol/L, (4.06 +/- 0.63) mmol/L. The expression of resistin in adipose tissue of obese rat fed with high fat diet was increased as compared with those fed with basic diet. CLA might increase the expression of resistin and PPARgamma in adipose tissue of obese rat.

CONCLUSIONThe expression of resistin mRNA of obese rat fed with high fat diet was higher than those fed with basic diet, and CLA might improve the insulin resistance in obese rats and possibly upregulate the expression of resistin through activing PPARgamma.

Adipose Tissue ; drug effects ; metabolism ; Animals ; Dietary Fats ; administration & dosage ; Gene Expression ; drug effects ; Insulin Resistance ; Linoleic Acids, Conjugated ; pharmacology ; Male ; Obesity ; etiology ; genetics ; PPAR gamma ; genetics ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Resistin ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVEIn order to accumulate experiences for improving the efficiency in serological tests, the present study on mixed testing of serum samples was performed by taking the serological test of trichinellosis and toxoplasmosis as the examples, and had proved the effects on cost-effectiveness of seroepidemiological survey of parasitic disease with method of mixed-samples test.

METHODSAccording to the binomial distribution principle, to develop an approach to the feasibility of mixed testing of serum samples, and to work on a cost-effectiveness analysis of one-by-one testing and mixed testing using hygienic economic analysis method was performed. For serological test of trichinellosis and toxoplasmosis, 3 kinds of mixed testing methods, namely 3 serum sample mixture, 5 serum sample mixture and 10 serum sample mixture, were performed.

RESULTSThe results showed that all the 3 kinds of mixed tests of trichinellosis and toxoplasmosis showing positive result if only 1 weak positive serum sample were mixed with. When the serum samples being mixed were all negative ones, then among the 24 groups tested with each kind of negative serum sample mixture of trichinellosis (3 serum samples, 5 serum samples and 10 serum samples), they all showed negative. However, among the 12 groups tested with 2 kinds of negative serum mixture of toxoplasmosis (3 serum samples and 5 serum samples), all showed negative while among the 18 groups tested with the 10 serum sample mixture, 16 groups showed negative and 2 were positive. The mixed testing of trichinellosis and toxoplasmosis showed that the efficiency of mixed testing was related to the serological positive rate of the parasitic diseases to be examined. When serological positive rate was 10%, the efficiency of mixed testing was higher in 4 serum sample group. When serological positive rate was 1%, the efficiency of mixed testing was higher in 10 serum sample group and when serological positive rate was 0.1%, the in crease of the size of mixed serum samples could decrease the number of testing, but the prerequisite was that there must be one positive sample, so that the positivity for all the mixed tests could be detected. If mixed testing were performed on all negative samples, no positivity could be detected.

CONCLUSIONThe result of cost-effectiveness analysis demonstrated that for seroepidemiological survey of parasitic diseases, the cost for mixed testing was low, especially when the serological positive rate was expected low (< or = 1%, thus the mixed testing could save a large amount of the cost.

Cost-Benefit Analysis ; Data Collection ; Humans ; Seroepidemiologic Studies ; Specimen Handling ; Toxoplasmosis ; diagnosis ; epidemiology ; Trichinellosis ; diagnosis ; epidemiology

OBJECTIVETo explore the clinical characteristics, diagnosis and surgical treatment of adult congenital bronchoesophageal fistula.

METHODSEleven cases of adult congenital bronchoesophageal fistula that were diagnosed and surgically treated between May 1990 and August 2010 had been reviewed. There were 7 male and 4 female patients, ranging in age from 28 to 66 years (mean 48.7 years). The chief clinical presentation included coughing and sputum in 10 cases, recurrent bouts of coughing after drinking liquid food in 6 cases, hemoptysis in 6 cases, low fever in 4 cases, chest pain in 3 cases. The duration of symptoms before diagnosis ranged from 5 to 36 years (mean 16.8 years). The diagnosis of bronchoesophageal fistula was confirmed most by esophagography. Associated diseased lung was resected in all patients (lobectomy in 10 cases and pneumonectomy in 1 case). The operation included right thoracotomy in 7 cases and left thoracotomy in 4 cases. The fistula was completely resected in 10 cases. The tract was simply divided and the end was sutured in 1 case.

RESULTSThe postoperative course was uneventful in 10 patients who were discharged from hospital 10 to 18 d after operation. One patient suffered from esophageal fistula and received second operation. Regular follow-up was conducted on all 11 patients, proving that 3-year survival rate was 11/11 and 5-year survival rate was 9/11.

CONCLUSIONPersistence of congenital bronchoesophageal fistula into adulthood is rare. The main symptom is nonspecific coughing and bouts of coughing after drinking liquid food. The most useful diagnostic method is the esophagography. Even though it is benign disease, life-threatening complications might occur and it must be treated surgically as soon as the diagnosis is established.

Adult ; Aged ; Bronchial Fistula ; congenital ; diagnosis ; surgery ; Esophagoscopy ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Survival Rate ; Thoracotomy

OBJECTIVETo investigate the constituents of Elsholtzia blanda.

METHODThe chemical components were isolated by polyamide and silica gel column chromatography. Their structures were identified with extensive spectral (EI-MS, ESI-MS, 1H-NMR, 13C-NMR, DEPT, 1H-1H-COSY, HMBC, HMQC) and chemical methods.

RESULTFive compounds were isolated and identified as luteolin (I), luteolin-5-O-beta-D-glucopyranoside (II), luteolin-7-O-beta-D-glucopyranoside (III), 5-hydroxy-7, 8 -dimethoxyflavone (IV) and 5-hydroxy-6, 7-dimethoxyflavone (V).

CONCLUSIONCompounds III, IV, V were isolated from E. blanda for the first time and I was firstly separated from the genus Elsholtzia.

Flavonoids ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Lamiaceae ; chemistry ; Luteolin ; chemistry ; isolation & purification ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVETo investigate the value of application of mandibular outer cortex as bone graft by comparing its bone absorption with cranial outer cortex.

METHODS8 minitype grown-up pigs at the age of 8 - 12 months underwent surgery of taking out the same size (2.5 cm x 1.0 cm) of outer cortex from mandible and craninium. The volume of the outer cortex was measured by volume-displacement method. Then the outer cortex of mandible and cranium were onlay grafted to the each side of the pig snout, respectively. 12 weeks later, 2 pigs were randomly selected for histological examination. The other 6 pigs were killed 24 weeks after surgery for measurement of the bone graft volume and histologic examination.

RESULTSThe bone graft absorption rate was (41 +/- 5)% for mandibular outer cortex and (46 +/- 12)% for cranial outer cortex, showing no significant difference between them (P = 0.51). The histologic examination results also had no marked difference in the bony healing and reforming between the two graft.

CONCLUSIONSMandibular outer cortex is a good donor site for onlay bone graft in craniofacial region.

Animals ; Bone Plates ; Bone Regeneration ; Bone Transplantation ; methods ; Female ; Male ; Mandible ; transplantation ; Skull ; transplantation ; Swine ; Swine, Miniature

OBJECTIVETo express endotoxin binding peptide and its mutant in E coli DH5alpha and detect the antiendotoxin activity of the purified peptides.

METHODS(1 ) E coli DH5at containing pinpointXa3-EBP and pinpointXa3-mEBP was activated by IPTG to express biotin fusion protein. The fusion proteins were purified, and then digested by factor Xa for isolation of EBP and mEBP. The target peptide was purified with affinity chromatography and reversed-phase HPLC. Sequences of 10 amino acids at N-terminal were used for identification of mEBP. (2) PBMCs were isolated from blood of normal people, and they were stimulated with 5 mg/L FITC-LPS plus 2.0,5. 0 and 12. 5 mg/L EBP or mEBP. Then the mean fluorescent intensity was detected. PBMC was also stimulated with 1 mg/L LPS plus 2.0, 5.0 and 12.5 mg/L EBP or mEBP for 5 hours for the detection of the TNF-alpha and IL-6 level in the supernatant. (3) Thirty-five Kunming mice were randomized into normal control ( n = 5, with intraperitoneal injection of 0. 2 ml isotonic saline) , model group(n = 5, with intraperitoneal injection of LPS and 20% TBSA full-thickness burns), and treatment group (n = 15, with intraperitoneal injection of 5 mg/kg PMB or 10 mg/kg EBP or mEBP after burns). The serum contents of TNF-a and IL-6, and TNF-alpha mRNA level in hepatic tissue in each group were determined 6 hours after treatment.

RESULTS( 1 ) EBP and mEBP were obtained after Xa digestion of biotin fusion protein, with purity reaching above 98% . The sequence of 10 amino acid at N-terminal was in accord with what expected. (2) The fluorescent intensity was decreased followed by an increase in mEBP or EBP concentration. Compared with normal PMBC, IL-6 and TNF-alpha level in the supernatant were obviously lowered in 1 mg/L LPS + 12.5 mg/L EBP group and I mg/L LPS +2. O0 , 5. 0, 12. 5 mg/L mEBP groups ( P < 0.01). (3) The serum level of IL-6 and TNF-ca in the therapeutic groups were obviously lower than that in model group ( P < 0.01 ) , and the levels of these cytokines were significantly lower in 10 mg/kg mEBP group than that in 10 mg/kg EBP group ( P <0. 01) , but they were similar to that in 5 mg/kg PMB treatment group ( P >0.05). (4) Relative optical density of TNF-alpha. mRNA in control, model, 10 mg/kg mEBP, 10 mg/kg EBP and 5 mg/kg PMB groups was 0.25, 0.93, 0.51 , 0.77 and 0.43, respectively.

CONCLUSIONEndotoxin binding peptides can be obtained by procaryon expression. Both EBP and mEBP have anti-LPS activity, but mEBP is more effective.

Animals ; Endotoxins ; metabolism ; Gene Expression ; Humans ; Interleukin-6 ; metabolism ; Lipopolysaccharides ; metabolism ; Membrane Proteins ; metabolism ; Mice ; Mice, Inbred Strains ; Mutant Proteins ; isolation & purification ; metabolism ; Peptides ; isolation & purification ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism