OBJECTIVE:To establish a method for the content determination of muscone in Aiweixin oral liquid. METHODS:After extracting sample by butanol,GC-MS was performed on the column of BD-17 MS with the volume temperature of 270 ℃;the carrier gas was helium with the sample size of 1 μl at the flow rate of 10 ml/min;the split ratio was 20∶1. EI was ion source with the electron energy of 70 eV,ion source temperature of 230 ℃ and quadrupole temperature of 150 ℃;the tuning mode was automatic tuning by quality full scanning with the threshold value of 30 in the range of 30-600 aum. RESULTS:The linear range of muscone was 12.216-61.08 ng(r=0.999 6);RSD of precision test was lower than 2% and the RSDs of reproducibility and stability tests were lower than 3%;the average recovery was 100.24%(RSD=1.58%,n=6). CONCLUSIONS:The method is sensitive, rapid and simple,and can be used for the content determination of muscone in Aiweixin oral lipid.

Hypoxia is a general characteristic of most solid malignancies and intimately related to cancer progression. Homeostatic response to hypoxia is primarily mediated by hypoxia inducible factor-1alpha (HIF-1alpha) that elicits transcriptional activity through recruitment P300 coactivator. Targeting the interaction of HIF- alpha and P300 would thus constitute a novel approach for cancer treatment by suppressing tumor angiogenesis and metastasis. Here, a screening assay was developed for inhibitors targeting the interaction between HIF-1alpha and P300. The nucleotide sequence of human HIF-1alpha and P300 were cloned into pBIND and pACT vectors, named pBIND-HIF1alpha and pACT-P300. The interaction of HIF-1alpha and P300 was identified in HEK293 cell using mammalian two-hybrid system. And compound chetomin decreased their interaction in this mammalian two-hybrid system. We further verified HIF-1 inhibition effect of chetomin in U251-HRE cells. Therefore, we established a screening assay combined HIF-1alpha and P300 mammalian two-hybrid system and U251-HRE reporter assay for HIF-1 selective inhibitors.
Cell Hypoxia ; Disulfides ; pharmacology ; Drug Screening Assays, Antitumor ; E1A-Associated p300 Protein ; antagonists & inhibitors ; HEK293 Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; antagonists & inhibitors ; Indole Alkaloids ; pharmacology ; Two-Hybrid System Techniques

OBJECTIVETo assess the association between SIRT1 gene polymorphisms and the longevity phenomena in Yongfu region of Guangxi. In this case-control study, 500 individuals from Yongfu region of Guangxi were recruited. The subjects were divided into a longevity group (n=223, average age=93.17 U+00B1 3.08 yr) and a healthy control group (n=277, average age=46.92 U+00B1 17.12 yr). Polymerase chain reaction-high resolution melting curve (PCR-HRM) and DNA sequencing were used to determine the allelic and genotypic frequencies of rs3758391, rs3740051, rs2273773, rs4746720 and rs10997870 polymorphisms of SIRT1 gene in the two groups. The association between above polymorphisms and longevity was assessed.

RESULTSIn the longevity group, CT genotype of the rs4746720 locus was significantly more common than CC and TT genotypes (P=0.000, OR=2.098, 95%CI:1.412-4.117). However, no significant difference was found in the allelic and genotypic frequencies of rs3758391, rs3740051 and rs2273773 between the two groups.

CONCLUSIONThere is an association between rs4746720 of SIRT1 gene and longevity in Yongfu region of Guangxi.

Adult ; Aged ; Aged, 80 and over ; Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; China ; Female ; Gene Frequency ; Gene Order ; Genetic Association Studies ; Genotype ; Humans ; Longevity ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Sirtuin 1 ; genetics ; Young Adult

Objective:To improve the genetic stability of HBV gene in transgenic mice.Methods:HBV transgenic mice were bred by backcross and double cross.The HBV gene expression and replication were studied with real-time PCR,ELISA and chemiluminescence.Results:The HBV transgenic mice have stably bred to 23rd generation.The serum HBsAg level is 4122.31?2044.74IU/ml;The rate of HBV transgenic mice whose serum HBV DNA reach 104~106copies/ml was 93.93%.The HBV replication and expression were improved markedly.There is no difference between male and female mice about serum HBsAg level.Conclusion:After breeding the HBV gene was expressed stably with high-level in transgenic mice.

The purpose of this study is to investigate the activity and mechanism of a new anti-tumor agent T03. MTT and colony formation assay were performed to determine anti-proliferation activity of T03 in vitro. Antitumor activity was observed by Renca xenograft model in vivo. The effect of T03 on cell cycle and apoptosis were measured by FCM analysis. Western blotting was performed to investigate the expression level of proteins in HepG2 cell lines treated with T03. T03 had anti-tumor activity by inhibiting tumor cell growth and colony formation in vitro, especially on hepatocellular carcinoma cells (HCC). At the concentration of 10 micromol x L(-1), T03 induced cell apoptosis and cell cycle arrest in HCC. Moreover, it proved that T03 reduced the tumor weight with the rate of 42.30% without any obviously side effect in Renca xenograft model. At the concentration of 2.0 micromol x L(-1), T03 was able to reduce the level of p-c-Raf (Ser259), and thus blocked Raf/MEK/ERK and AKT signaling in HepG2 cell lines. The result suggested that T03 has the potential to inhibit cell proliferation and induce cell apoptosis both in vitro and in vivo, particularly active against HCC, indicating T03 and its analogues may serve as a new anti-cancer drug against hepatocellular carcinoma.
Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Cell Cycle Checkpoints ; drug effects ; Cell Proliferation ; drug effects ; Hep G2 Cells ; drug effects ; Humans ; Liver Neoplasms ; pathology ; Signal Transduction ; drug effects ; Xenograft Model Antitumor Assays

Objective To evaluate the clinical efficacy and safety of bi-calutamide combine with goserelin in the treatment of advanced prostate cancer.Methods Fifty cases of advanced prostate cancer patients were randomly divided into treatment group (25 cases) and control group (25 cases).Treatment group was given oral bicalutamide 50 mg, qd +sub-cutaneous goserelin 3.6 mg, once 4 weeks, a monthly review of serum prostate-specific antigen ( PSA ) content , when the serum PSA <0.2 ng· mL-1, disabled two drugs, when PSA>4 ng· mL-1, begin a new round of treatment.Control group was given the continuous dosing , the same dosage and usage as the treatment group.The course of treatment for two groups was 12 months.Comparison of the clinical efficacy , serum PSA value and adverse drug reactions.Results After 6, 9, 12 months of treatment , the clinical efficacy of treatment group was significantly higher than that of control group (P<0.05).After 3, 6, 9, 12 months of treatment , the serum PSA value was significantly lower than that of before treatment ( P<0.05 ) , and there was no significant difference in serum PSA between the two groups at each time point ( P>0.05 ).The main adverse drug reactions of the two groups were the castration syndrome , blood vessel contraction , osteoporosis , and liver toxicity, and the incidence rate of adverse drug reactions were not statistically different between the two groups ( P>0.05 ).Conclusion The clinical efficacy of intermittent administration for bicalutamide combined with goserelin in the treatment of advanced prostate cancer was significantly better than the continuous dosing , without increasing the incidence of adverse drug reactions.

OBJECTIVETo investigate the correlation of the common variant single nucleotide polymorphisms (SNP) on chromosome 3 with the incidence and related risk factors of prostate cancer (PCa) in Chinese men.

METHODSUsing the case-control meth- od, we included 124 PCa patients in the PCa group and 111 age- and gender-matched cancer-free healthy subjects as normal controls. We detected the distribution of allele and genotype frequencies of the SNP rs10934853 and rs2660753 with the polymerase chain reaction-high resolution melting curve (PCR-HRM) combined with gene sequencing, analyzed the cumulative effect of the risk genotypes of these two independent variants, and determined the correlation between different genotypes of these two SNPs and clinically related risk factors in the PCa patients.

RESULTSAs for the genotypes of rs10934853, there were 28 cases of AA (22.8%), 46 cases of CC (37.4%), and 49 cases of AC (39.8%) in the PCa patients, as compared with 24 (22.0%), 34 (31.2%) and 51 (46.8%) in the healthy controls. As regards the genotypes of rs2660753, there were 13 cases of AA (11.0%), 59 cases of GG (50.0%) and 46 cases of AG (39.0%) in the PCa patients, in comparison with 9 (8.8%), 47 (45.6%) and 47 (45.6%) in the controls. No significant differences were found in the distribution of the genotype and allele frequencies of rs10934853 and rs2660753 between the two groups (P = 0.520 & 0.582). Analysis on the cumulative effect of the risk genotypes of rs10934853 and rs2660753 showed a slightly higher risk of PCa (OR = 1.831 & 1.968) in the two groups with risk genotypes than in the one with wild types (P > 0.05). Different genotypes of rs10934853 and rs2660753 were not correlated with clinically related risk factors of the PCa patients (P > 0.05).

CONCLUSIONSNP rs10934853 and rs2660753 on chromosome 3 are not obviously correlated with PCa in Chinese patients, and may not be a genetic risk factor of PCa.

Aged ; Aged, 80 and over ; Alleles ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Chromosomes, Human, Pair 3 ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Prostatic Neoplasms ; epidemiology ; genetics ; Risk Factors

OBJECTIVETo explore the role of cytokines and keratinocytes in the survival mechanism of mixed auto and allogeneic skin grafting.

METHODSThirty-six SD rats were employed in the study. The rat model with mixed auto and allogeneic skin grafting and mixed human epithelial and lymphocytic culture (MELC) model were established. The change of IL-10 in the serum and the supernatant of the cultured tissue sample from the local wound was observed after the mixed skin grafting in scalded rats. And the role of epithelium in the induction of immunosuppression in vitro was monitored.

RESULTSThe serum IL-10 content in the rats with mixed skin grafting (25.89 +/- 2.82 ng/L) at 7 postoperative day (POD) was evidently higher than that in normal rats (14.20 +/- 2.43 ng/L) (P < 0.05). The IL-10 content in the culture supernatant of rat tissue samples exhibited evident different during 4-14 PODs (P < 0.05-0.01), while which was no difference to that in normal rat on 21st and 28th POD. The inhibiting effects of autologous epithelia and keratinocytes in MELC system were correlated with their dosage. After the adding of autologous keratinocytes to MELC system the cytokines secreted from Th1 could induce the secretion of cytokines from Th2 by IL-10 mediation. This effect could be corrected by the addition of monoclonal antibody of IL-10.

CONCLUSIONThe keratinocytes inlayed in the autoskin during mixed grafting could increase the local IL-10 level by activating Th2 cells, which might be one of the important reasons of the survival of mixed skin grafting.

Animals ; Burns ; immunology ; surgery ; Cytokines ; metabolism ; Giant Cells, Langhans ; cytology ; Graft Survival ; immunology ; Humans ; Interleukin-10 ; metabolism ; Keratinocytes ; cytology ; Lymphocyte Culture Test, Mixed ; Male ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; immunology ; Th2 Cells ; metabolism ; Transplantation, Autologous ; Transplantation, Homologous

Novel reassortant H3N2 swine influenza viruses (SwIV) with the matrix gene from the 2009 H1N1 pandemic virus have been isolated in many countries as well as during outbreaks in multiple states in the United States, indicating that H3N2 SwIV might be a potential threat to public health. Since southern China is the world's largest producer of pigs, efficient vaccines should be developed to prevent pigs from acquiring H3N2 subtype SwIV infections, and thus limit the possibility of SwIV infection at agricultural fairs. In this study, a high-growth reassortant virus (GD/PR8) was generated by plasmid-based reverse genetics and tested as a candidate inactivated vaccine. The protective efficacy of this vaccine was evaluated in mice by challenging them with another H3N2 SwIV isolate [A/Swine/Heilongjiang/1/05 (H3N2) (HLJ/05)]. Prime and booster inoculation with GD/PR8 vaccine yielded high-titer serum hemagglutination inhibiting antibodies and IgG antibodies. Complete protection of mice against H3N2 SwIV was observed, with significantly reduced lung lesion and viral loads in vaccine-inoculated mice relative to mock-vaccinated controls. These results suggest that the GD/PR8 vaccine may serve as a promising candidate for rapid intervention of H3N2 SwIV outbreaks in China.
Animals ; Female ; Influenza A Virus, H3N2 Subtype/*genetics/immunology ; Influenza Vaccines/genetics/immunology/*therapeutic use ; Mice ; Mice, Inbred BALB C ; Orthomyxoviridae Infections/immunology/*prevention & control/virology ; Reassortant Viruses/genetics/immunology ; Reverse Genetics/methods/*veterinary ; Swine ; Swine Diseases/immunology/*prevention & control/virology ; Vaccines, Inactivated ; Virus Replication

In European populations, 7 single nucleotide polymorphisms (SNPs) on chromosome 17q, 3 SNPs on 17q12, and 4 SNPs on 17q24.3 were recently identified to be closely related to the risk of prostate cancer by a genome-wide association study. In Japanese populations, the correlation between 2 SNPs on 17q and the risk of prostate cancer and tumor aggressiveness was also confirmed by a large-scale experiment. However, whether 17q is associated with prostate cancer and its clinical manifestations in Chinese populations is still unknown. Therefore, we conducted a case-control study in a northern Chinese population and tested 2 SNPs, rs4430796 and rs1859962, on 17q in 124 prostate cancer patients and 111 controls using polymerase chain reaction-high resolution melting curve (PCR-HRM) combined with sequencing. We analyzed the association of the 2 SNPs with the risk of prostate cancer as well as patients' lifestyles, onset ages, Gleason scores, PSA levels, and pathologic stages. We found a significant difference in the G allele of SNP rs1859962 (P = 0.035, OR = 1.51, 95% CI = 1.03-2.21) but not in the rs4430796 genotype frequency or allele frequency distribution between prostate cancer patients and the controls (P > 0.05). Neither of the SNPs was significantly associated with the onset age, Gleason score, PSA level, pathologic stage, or other clinical indicators of patients with prostate cancer (P > 0.05). Our results show that polymorphism of the G allele of SNP rs1859962 is associated with the risk of prostate cancer in a Chinese population.
Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Chromosomes, Human, Pair 17 ; genetics ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Neoplasm Grading ; Neoplasm Staging ; Polymorphism, Single Nucleotide ; Prostatic Neoplasms ; genetics ; Risk Factors