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

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.

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.

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

Angioplasty ; Embolectomy ; Ischemia ; pathology ; surgery ; Mesenteric Artery, Superior ; pathology ; surgery ; Mesenteric Ischemia ; Thrombosis ; pathology ; surgery ; Vascular Diseases ; pathology ; surgery

OBJECTIVETo objectively assess the clinical efficacy and safety of a new Pulian Ointment (, NPLO) in treating psoriasis of blood-heat syndrome of Chinese medicine.

METHODSA total of 108 patients with psoriasis of blood-heat syndrome were equally assigned, using a randomizing digital table, to the test group treated externally with NPLO and the control group treated with placebo; the medication was done using a singleblinded method twice a day. Meanwhile, all patients received by oral intake a conventional Chinese decoction for clearing heat and cooling blood; the therapeutic course was 4 weeks for both groups. The therapeutic efficacy, changes in the Psoriasis Area and Severity Index (PASI) score and various aspects of the lesion, including scaly eruption, erythema, infiltration, size, score of itching as well as adverse reactions were observed.

RESULTSThe trial was completed in 100 patients, 51 in the test group and 49 in the control group. The remarkably effective rate was 45.10% and the total effective rate was 84.31% in the test group, which were significantly higher than those in the control group, 12.24% and 51.02%, respectively, showing a significant difference between groups (P<0.01). The test group also showed better effects in the improvement of the PASI score of the lesions and scores on erythema, infiltration, size of lesion as well as itching. No adverse event was found in either group.

CONCLUSIONNPLO is a Chinese remedy for the external treatment of psoriasis of the blood-heat syndrome with a reliable therapeutic efficacy and good safety.

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Ointments ; Psoriasis ; drug therapy ; Syndrome

OBJECTIVETo identify possible reasons of rotational mismatch between tibial component and femoral component in total knee arthroplasty and choose a right reference axis for placing the components in the operation and to decrease the complications.

METHODSForty normal Chinese knees were studied. There were 20 men and 20 women, and average age was 34 years (range, 18 - 42 years). The images of cross sections of the distal femur were obtained by spiral CT scanning (0.5 mm thickness). Scan direction was aligned to be in the plane perpendicular to the mechanical axis of the tibia. On the images of the distal femur and the proximal tibia, three baselines for the anteroposterior axis of the femoral component were drawn based on the clinical epicondylar axis and the surgical epicondylar axis and 3 degrees lateral rotated to the posterior condylar surfaces of the femur separately, and a baseline for the anteroposterior axis of the tibial component was drawn based on the medial 1/3 of the tibial tuberosity. The rotational mismatch angles were measured between each component by using the Autocad software.

RESULTSThe mean rotational mismatch angle between tibial component and femoral component is 2.94 degrees for the clinical epicondylar axis, 6.50 degrees for the surgical epicondylar axis and 6.83 degrees for 3 degrees lateral rotation of the femoral component referenced to the posterior condylar axis separately.

CONCLUSIONSLandmarks of each bone were the intrinsic cause of the rotational mismatch in total knee arthroplasty. The clinical epicondylar axis can be chosen for the ideal reference to rotational alignment of the femoral component because of its minimal rotational mismatch between each component.

Adolescent ; Adult ; Arthroplasty, Replacement, Knee ; methods ; Female ; Humans ; Knee Joint ; diagnostic imaging ; physiopathology ; surgery ; Male ; Postoperative Complications ; prevention & control ; Range of Motion, Articular ; Tomography, Spiral Computed

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

OBJECTIVETo study the relationship between influenza epidemic and genetic characteristic on HA and NA regions of influenza virus subtype A3 isolates of Zhejiang province in the recent years.

METHODSRNA of 25 influenza virus subtype A3 isolates, circulated in Zhejiang province during 1998 to 2005, was extracted. HA1 and NA regions were amplified and sequenced. All the sequence data were analyzed using BioEdit.

RESULTSHA1 and NA regions of all the isolates belonged to 987nt and 1362nt, encoding protein of 329 and 454 amino acids respectively. Isolates shared amino acid homology of 90.9%-99.3% and 95.2%-99.5% on HA1 and NA regions, while divergence on HA1 was greater than that on NA region. During a period of 8 years, 30 amino acids on HA1 region were substituted and 14 of which refer to 4 antigenic determinant sites. Meanwhile,21 amino acids on NA region were substituted and 5 of which referred to 3 antigenic determinant sites. Significant divergences, both in HA1 and NA, were observed among isolates in 1998 and 2002, showing that they belonged to absolutely different branches. Additionally, influenza virus subtype A3 isolates identified in recent years, with 11 N-linked glyeosylation sites in HA1 region, had 5 sites more than early A/Aichi/2/68 strain. Since 1998,3 sites had been inserted in epidemic strains, indicating the accelerated trend of glyeosylation sites were increasing.

CONCLUSIONThere is a correlation between antigenic drift of influenza virus subtype A3 and the two epidemics in Zhejiang province in 1998 and 2002.

Amino Acid Sequence ; Antigens, Viral ; genetics ; China ; Epitopes ; genetics ; Evolution, Molecular ; Genetic Drift ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A virus ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; Molecular Sequence Data ; Neuraminidase ; genetics ; RNA, Viral ; genetics ; Sequence Analysis, RNA ; Sequence Homology, Amino Acid