Objective: To investigate the analgesic substances in the aerial part of Urtica fissa (Urticae Fissae Herba), commonly used for rheumatoid and rheumatism arthritis. Methods: The analgesic constituents were isolated with the active guidance of hot plate and acetic acid writhing models, and identified by comprehensive spectroscopic analysis. Results: Thirteen alkaloids (1–13), two lignans (14, 15), and three amides (16–18) were isolated from the active fractions. Among them, compound 1 was a new alkaloid, and compound 6 was a new natural product. The activity evaluation in vivo indicated that various pyrrole alkaloids (1, 3, 6, and 12) possessed significant analgesic activities, they could significantly inhibit the mice pain response induced by acetic acid and hot plate at the dosage of 2 mg/kg BW. Conclusion: The study revealed that the pyrrole alkaloids played important roles in the analgesic activities of Urticae Fissae Herba.

BACKGROUND: The association of genetic variation in the IRAK-1 gene with sepsis outcome has been proved. However, few studies have addressed the impact of the IRAK-4 gene variants on sepsis risk. This study aimed to determine whether the polymorphisms in the IRAK-4 gene are associated with susceptibility to and prognosis of severe sepsis in the Chinese Han ethnic population.METHODS: In this case-control study, 192 patients with severe sepsis hospitalized in the emergency department of Zhongshan Hospital from February 2006 to December 2009 and 192 healthy volunteers were enrolled. Exclusion criteria included metastatic tumors, autoimmune diseases, AIDS or treatment with immunosuppressive drugs. This study was approved by the ethical committee of Zhongshan Hospital, Fudan University. Sepsis patients were divided into a survival group (n=124) and a non-survival group (n=68) according to the 30-day mortality. Primer 3 software was used to design PCR and sequencing primers. Genomic DNA was extracted from peripheral blood mononuclear cells. Seven tagSNPs in IRAK-4 were selected according to the data of the Chinese Han population in Beijing from the Hapmap project and genotyped by direct sequencing. The chi-square test was used to evaluate the differences in genotype and allele frequencies between the two groups.RESULTS: The distributions of all tagSNPs were consistent with Hardy-Weinberg equilibrium. The allele and genotype frequencies of rs4251545 (G/A) were significantly different between the severe sepsis and healthy control groups (P=0.015, P=0.035, respectively). Carriers of the rs4251545A had a higher risk for severe sepsis compared with carriers of the rs4251545G (OR=1.69, 95％ CI: 1.10-2.58). The allele and genotype frequencies of all SNPs were not significantly different between the survival group and non-survival group.CONCLUSION: These findings indicate that the variants in IRAK-4 are significantly associated with susceptibility to severe sepsis in the Chinese Han ethnic population.

OBJECTIVETo investigate the relationship between the androgen receptor (AR), oestrogen receptor( ER) and progesterone receptor (PR) by assaying the distributions of these three receptors in the both benign prostate hyperplasia (BPH) and prostatic cancer (PCa) tissues, then to explore the difference of the effects of hormonal treatment between the AR positive and AR negative in PCa patients.

METHODSTo observe AR, PR and ER expressions in 30 cases of BPH and 32 cases of PCa tissues by DAKO Envision methods.

RESULTSThere were 22 AR positive, 10 ER positive and 6 PR positive cases in 30 BPH patients, and there were 18 AR positive, 14 ER positive and 2 PR positive in 32 PCa patients. There was no significant difference between the two groups. The survival time was 6.41 and 4.28 years for the AR positive and AR negative respectively in 30 PCa patients. The AR positive patients lived longer than the AR negative (P < 0.05).

CONCLUSIONThese three receptors cant result in PCa and the effects of hormonal treatment on AR negative PCa patients is not certain.

Aged ; Aged, 80 and over ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Prostatic Hyperplasia ; metabolism ; Prostatic Neoplasms ; metabolism ; Receptors, Androgen ; biosynthesis ; Receptors, Estrogen ; biosynthesis ; Receptors, Progesterone ; biosynthesis

A new method for the preparation of oligonucleotide microarray for gene expression detection was found. The key techniques and standards of quality controlling for preparation of oligonucleotide microarray was explored using gene of human 23 kD highly protein and Luciferase and mouse cytokine-associated genes. By the using of a software system MProbe, oligonucleotide probes were designed and BLAST. All the probes have a very high specificity, i.e. except target sequence, the similarity between the probe and non-target sequences is less than 70% and the hairpin structure are not exist in all probes. All the probes have the same length 40. GC contents in all probes are in a narrow scope (from 45% to 55%). All the probes are modified with amino at 5' or 3' terminal. The satisfied images with good sensitivity and very high specificity were obtained by the using of the methods above and also using of positive and negative controls and some internal controls(house keeping gene) to quantitate and balance expression of genes. High specificity, good sensitivity and stability have been verified by three continuous experiments using the oligonucleotide microarray to study gene expression profile of normal mouse breast grand tissue. The oligonucleotide microarray for expression detection prepared using our method have high specificity, good sensitivity and stability et al. It may be a more advanced method for analysis of gene expression profile.
Animals ; Cytokines ; genetics ; DNA, Complementary ; genetics ; Gene Expression Profiling ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; Interleukin-10 ; genetics ; Mammary Glands, Animal ; metabolism ; Mice ; Nerve Growth Factor ; genetics ; Oligonucleotide Array Sequence Analysis ; methods ; Polymerase Chain Reaction

Objective: To investigate the influence of CD133

OBJECTIVETo preserve the function of the donor site and good cervical shape, a modified pectoralis major myocutaneous island flap was designed.

METHODSThe modified pectoralis major myocutaneous flaps were used to repair primarily the defect in head and neck surgery. In all 17 cases, six cases were patients with recurrence of larynx or hypopharynx cancer, four cases with hypopharynx cancer, three cases with base of tongue cancer, two cases with recurrence of maxillary cancer, one case with tonsillar cancer and one case with pharyngeal fistula after hypopharyngeal cancer surgery. Before operation, ultrasound was used to mark the projection of the pectoral branches of thoracoacromial artery, and the pectoralis major myocutaneous were designed according to the axle between lowest entering muscle point of the artery and the fourth intercostals perforator spot of mammary artery; the incision was designed to turn laterally in an oriental direction at the top of the flap and upward along the anterior axillary line; the internal pectoral nerve was reserved, as well as the partial lateral pectoral nerve. The flaps were transferred to recipient site either above or below the clavicle on the premise of the integrity of clavicular part.

RESULTSThe distance of the lowest entering muscle point of pectoral branche measuring during operation, which was all in sternocostal part, to the midpoint of inferior clavicula margin was (4.9 +/- 1.2) cm (average +/- s), and in 76.5% (13/17) of the patients, the location was coincidence by ultrasound. The length between entering muscle point and the fourth intercostals perforator spot of mammary artery was (1.8 +/- 0.5) cm. All the myocutaneous flaps were alive except one case. The flap was given up as a result of the vessel pedicle injure. The distal end of the flap was dehisced from the residual tongue in one case with base of tongue cancer and healed with changing dressing. Two pharyngeal fistulas in another two cases were healed with conserved treatment. The rate of the flap survival was 94.1% (16/17). Functions as adduction and adtorsion of major pectoral muscle were integrated within 4 weeks to 3 months. Also, the good looking of the neck and upper chest was maintained.

CONCLUSIONSThe location of pectoral branches of thoracoacromial artery and the site of the lowest entering muscle point marked by ultrasound detection could help the design of the flap. The modified pectoral' s major myocutaneous flap designing presented better functional protection and reach longer distance and left a better looking for neck and upper chest.

Adult ; Aged ; Head ; surgery ; Head and Neck Neoplasms ; surgery ; Humans ; Male ; Middle Aged ; Neck ; surgery ; Pectoralis Muscles ; transplantation ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; Surgical Flaps

OBJECTIVETo investigate the pharmacological effects and underlying mechanism of azidothymidine (AZT) on human glioblastoma cells in vitro.

METHODSThe telomerase activity of human glioblastoma TJ905 cells was determined by TRAP assay after 24 hrs' incubation with 50, 100, 200 micromol/L AZT and control vehicle solution. Colony formation efficiencies of the cells were recorded. Cells of the 1st, 3rd and 6th generations were harvested, followed by evaluations of cyclin A protein expression by Western blot, cell cycle distribution by flow cytometry, apoptotic level by single cell gel electrophoresis and proliferation index by Ki-67 immunocytochemical staining.

RESULTSAZT inhibited telomerase activity of TJ905 cells. Cyclin A expression levels in the cells treated with 50 and 100 micromol/L AZT were significantly lower than controls (P < 0.01), and down-regulation of the expression was in a dose- and time-dependent manner. Compared with controls, G(0)/G(1) phase cells were obviously decreased (P < 0.05 approximately 0.01) and S phase cells significantly increased (P < 0.05 approximately 0.01) after treatment with 50, 100 and 200 micromol/L AZT. The cell numbers of G(0)/G(1) and S phases at the 1st generation of above three treated groups changed in a dose-dependent manner, whereas S phase cells increases in all AZT treatment groups and G(0)/G(1) phase cell decrease in group treated with 50 micromol/L AZT were also in a time-dependent manner. Both the apoptotic cells of the 1st and 6th generations of all AZT treatment groups were significantly more than controls (P < 0.05 approximately 0.01), their numbers of the 6th generations of the three groups increased with AZT concentration (P < 0.05 approximately 0.01), and all of them were more than the 1st and 3rd generations of the same dosage group (P < 0.05 approximately 0.01). Colony formation efficiencies and Ki-67 labeling indexes of the three AZT treatment groups were distinctly lower than controls (P < 0.01), and they were also decreased with the elevation of AZT concentration and/or the elongation of the incubating time. The difference of any above parameter had no significance among the 1st, 3rd and 6th generations of control group (P > 0.05).

CONCLUSIONAZT blocks S/G(2) conversion of TJ905 cells by inhibition of telomerase activity and cyclin A expression, leading to an enhancement of apoptosis and suppression of cell proliferation.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin A ; metabolism ; Dose-Response Relationship, Drug ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Glioblastoma ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Reverse Transcriptase Inhibitors ; administration & dosage ; pharmacology ; Telomerase ; metabolism ; Zidovudine ; administration & dosage ; pharmacology

BACKGROUNDLittle is known about the prognosis of coronary artery disease (CAD) in Chinese patients with abdominal aortic aneurysm (AAA). The aim of this study was to evaluate the predictors of in-hospital all-cause mortality of severe CAD in Chinese patients who were hospitalized for AAAs.

METHODSFrom January 2003 to August 2009, 368 patients were operated on for AAAs. The clinical characteristics were retrospectively collected. The primary outcome was the in-hospital all-cause mortality. The clinical risk factors were subjected to a multivariate analysis to determine the predictors of in-hospital all-cause mortality.

RESULTSDuring their hospitalization, 23% (85/368) of the patients underwent coronary angiography, which revealed significant lesions in 93% (79/85) of the patients. In 25 cases, coronary artery bypass grafting (CABG) was performed before the AAA repair and in 16 cases of percutaneous coronary intervention (PCI) was performed. Ten patients with AAA alone died before discharge, and eight patients diagnosed with AAA combined with CAD died. There was no statistical difference in the postoperative death between the two groups. The logistic analysis showed that age > 70 years and CAD (vessels ≥ 2) were the significant factors in predicting the adverse clinical outcome.

CONCLUSIONSThe prevalence of severe CAD in Chinese patients with AAAs seemed lower than those that were reported. Myocardial evaluation and subsequent revascularization before AAA surgery could improve the clinical outcome for these patients who have severe CAD.

Adult ; Aged ; Aged, 80 and over ; Aortic Aneurysm, Abdominal ; complications ; mortality ; surgery ; China ; epidemiology ; Coronary Artery Disease ; epidemiology ; mortality ; surgery ; Female ; Hospital Mortality ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Prevalence ; Retrospective Studies

To produce the recombinant baculovirus transfer plasmid pFast-ORF2, the ORF2 gene of Porcine Circovirus type 2 (PCV2) was subcloned into baculovirus transfer vector (pFastBac(TM1) ) using Bac-to-Bac baculovirus expression system. E. coli DH10Bac (Gibco BRL) containing baculovirus shutter vector (bacmid) and helper vector was transformed with recombinant plasmid pFast-ORF2. Within E. coli DH10Bac, the ORF2 gene was transposed into the bacmid. The colonies of E. coli containing recombinant bacmid (Bac. ORF2) were collected by blue/white selection. The Bac. ORF2 was transfected into sf9 cells to yield AcNPV carrying the PCV2 ORF2 gene, referred to as Ac. ORF2. Expression of the ORF2 gene of PCV2 was confirmed by indirect immunofluorescent assay (IIFA), SDS-PAGE and Western-blotting. The expressed ORF2 gene product had a molecular mass of 28kD and could be recognized by the positive serum of PCV2. The results indicated the ORF2 gene was properly expressed in sf9 cell. It was noteworthy that many self-assembled virus-like particles (VLPs) were found in purified and phosphotungstic acid (PTA) stained PCV2 ORF2 protein by electron microscope. The particles were of similar morphology to the PCV2 virion and some self-assembled virus-like particles had darkly stained centers that made them appear to be empty capsids. Both PCV2 particles and self-assembled particles were approximately 17 nm in diameter.
Animals ; Baculoviridae ; genetics ; metabolism ; Circovirus ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Insecta ; cytology ; metabolism ; Open Reading Frames ; genetics ; Recombinant Proteins ; genetics ; metabolism ; Swine ; Viral Proteins ; genetics ; metabolism ; Virion

Humans ; Transcranial Magnetic Stimulation ; Pain Management ; Psychotic Disorders/therapy* ; Depressive Disorder, Major ; Treatment Outcome ; Transcranial Direct Current Stimulation