Objective To investigate the inhibitory effect of sorafenib on the proliferation of cervical cancer HeLa cells and explore the mechanism of its antitumor mechanism.Methods Cervical cancer HeLa cells were treated with sorafenib at a final concentration of 0.01,0.1,1.0,2.5,5.0 mg · L-1 (experimental group).Control group was not given any drug treatment (HeLa cells).The proliferation of cervical cancer HeLa cells cultured for 1,2,3 d were detected by MTT assay.The expression of vascular endothelial growth factor-C (VEGF-C) and phosphorylated extracellular signal-regulated kinase (p-ERK) protein were detected by immunofluorescence assay and enzyme-linked immunosorbent assay (ELISA).Results After 3 d,the proliferation inhibition rates in 0.01,0.1,1.0,2.5,5.0 mg · L-1 experimental groups were (9.16 ± 1.58)％,(23.07 ± 1.86)％,(30.86 ± 2.67)％,(55.94 ± 3.92)％,(68.75 ±4.48)％,had significant difference with that in control group,which was (5.15 ± 1.67)％ (P ＜0.05).Immunofluorescence assay showed that,the green fluorescence in control group enhanced significantly,and the green fluorescence decreased significantly after treated with different concentrations of sorafenib.The expression of VEGF-Cin 0.01,0.1,1.0 mg L-1 experimental groups were (1305.64 ± 119.82),(1235.57 ± 102.34),(1083.33 ±89.61)pg · mL-1,had significant difference with that in control group,which was(1326.54 ± 139.57)pg· mL-1(P ＜0.05).The expression of p-ERK in 0.01,0.1,1.0 mg · L-1 experimental groups were (2.21 ±0.22),(1.56 ± 0.21),(0.96 ± 0.13) μg · mL-1,had significant difference with that in control group,which was(2.34 ± 0.28)μg · mL-1 (P ＜ 0.05).The expression of VEGF-C and p-ERK protein in HeLa cell culture supernatant decreased gradually with the increasing the sorafenib concentration (P ＜ 0.05).Conclusion Sorafenib can inhibit the proliferation of cervical cancer HeLa cells,and the proliferation inhibition rate is positively related to time and concentration.The mechanism of tumor suppressor may be related to its ability to inhibit the expression of VEGF-C and p-ERK protein in cervical cancer HeLa cells.

BACKGROUNDTo study the clinical features and more reasonable typing criteria for patients with chronic severe hepatitis and decompensated liver function.

METHODSData of 106 cases of decompensated cirrhosis, 124 cases of chronic liver failure and 100 cases of chronic liver failure (chronic liver failure group I, CLF I) with decompensated cirrhosis (chronic liver failure group II, CLF II) were analyzed retrospectively.

RESULTS(1) The ages were youngest in chronic liver failure group I (about 30 years), and the oldest in decompensated cirrhosis group (about 50 years). (2) There were significant differences in albumin, globulin, ALT, AST, protruding activity, blood glucose, blood lipid and cholinesterase among the three groups. (3) There was no significant difference in upper digestive tract bleeding and hepatorenal syndrome, on the other hand, there was significant difference in ascites and hepatic encephalopathy. (4) The prognosis of the patients in decompensated cirrhosis group was better than that of chronic liver failure group I and chronic liver failure group II.

CONCLUSIONThe clinical feature and prognosis in three groups were different, so, it is suggested that chronic severe liver disease be divided into 2 types: one is chronic severe liver disease type I, which is associated with chronic hepatitis, and the other is chronic severe liver disease type II, which is associated with cirrhosis, and the typing criteria for decompensated cirrhosis remains unchanged.

Adult ; Diagnosis, Differential ; Female ; Hepatitis, Chronic ; classification ; complications ; diagnosis ; Humans ; Liver Cirrhosis ; classification ; complications ; diagnosis ; Liver Failure ; etiology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies

It is unclear why the concentration of testosterone increases in the testicular vein after hemicastration without corresponding alteration in gonadotropins. The present work was undertaken to examine whether the testosterone levels could be modified by denervation of the testis in adult rats. Both systemic and testicular blood samples were collected either immediately before or 6 and 24 hours after hemicastration from the rats two weeks after denervation of either inferior spermatic nerves (ISN) or ISN plus superior spermatic nerves (ISN-SSN). Increase of testicular testosterone induced by hemicastration was significantly (P<0.05) inhibited in these rats, as compared with the sham animals (at 6 and 24 hours, ISN vs sham: 16.00±3.35 vs 42.72±13.85 and 26.93±8.68 vs 71.16±13.30 whilst ISN-SSN vs sham: 31.63±7.92 vs 60.61±18.11 and 27.70±8.93 vs 93.92±19.73 ng/ml, respectively), whereas no significant change in LH was observed in all the experimental groups. FSH underwent no alteration in all the ISN denervation groups, but a significant elevation was observed in the ISN-SSN denervation groups (P<0.05) before hemicastration. Therefore, it appears that the change in FSH is not the cause of the inhibition of testosterone increase in the hemicastrated rats after testicular denervation and that ISN plays an active role in regulation of testosterone increase induced by hemicastration.

OBJECTIVETo study the clinical feature and more reasonable diagnostic typing criteria for patients with liver failure.

METHODS13/21 cases of ALF, SALF with no past liver disease, 49/72 cases of with chronic hepatitis, and 23/73 cases ALF, SALF with liver cirrhosis, were analyzed respectively.

RESULTS1 ALF patients (1). There exist significant statistic differences in ALB, ALT, CHE in three ALF groups.(2). It had statistic differences in those patients with hepatic encephalopathy.(3). The prognosis of the patients with chronic hepatitis group (42.85 percent) was best than that of chronic cirrhosis (26.09 percent) and no past liver disease (15.38 percent). (2) In SALF patients (1). There exist significant statistic differences in ALB, GLO, ALT, AST, BDIL, GLU and CHE in three SALF groups.(2). It had statistic differences in those patients with hepatic encephalopathy in three SALF groups.(3). The prognosis of the patients with chronic hepatitis group (51.39 percent) was best than that of chronic cirrhosis (36.85 percent) and no past liver disease (33.33 percent).

CONCLUSIONThere are different clinic feature and prognosis in three ALF or SALF groups, so we suggest that it were clinic practicability and science in classify of liver failure at present.

Humans ; Liver Failure ; classification ; Liver Failure, Acute ; classification ; Prognosis

OBJECTIVESTo study the effects of trichostatin A (TSA) on protein-protein interaction between HBx and histone deacetylase protein 1 (HDAC1).

METHODSBoth HBx and HDAC1 expressing vectors were constructed by the method of routine molecular cloning. The expression of HBx and HDAC1 were observed by Western blot assay. The protein-protein interaction was tested between HBx and HDAC1 by GST pull-down in vitro as well as co-immunoprecipitation in vivo.

RESULTSBoth HBx and HDAC1 expressing vectors were successfully constructed. Protein-protein interaction between HBx and HDAC1 existed both in vitro and in vivo. TSA, an inhibitor of HDAC1, had no effect on the interaction between HBx and HDAC1.

CONCLUSIONSHBx interacts with HDAC1 in vivo and in vitro in a non- TSA dependent way.

Histone Deacetylase 1 ; metabolism ; Humans ; Hydroxamic Acids ; metabolism ; Immunoprecipitation ; Plasmids ; Protein Interaction Mapping ; Trans-Activators ; metabolism

Nine compounds were isolated from an ethanol extract of the roots of K. roxburghii by using a combination of various chromatographic techniques including column chromatography over silica gel, MCI gel, Sephadex LH-20, and reversed-phase HPLC. On the basis of physical-chemical properties and spectroscopic data analysis, their structures were identified as munjistin (1), 1-methoxy-3,6-dihydroxy-2-hydroxymethyl-9,10-anthraquinone (2), 1,2,3-trihydroxy-9,10-anthraquinone (3), arjunolic acid (4), hyptatic acid-A (5), hyptatic acid-B (6), 2α,3β,24-trihydroxyurs-12-en-28-oic acid (7), 2α,3β,23-trihydroxyurs-12-en-28-oic acid (8), and daucosterol (9). Compounds 1-9 were obtained from this genus for the first time.
Anthraquinones ; chemistry ; isolation & purification ; Rubiaceae ; chemistry ; Triterpenes ; chemistry ; isolation & purification

OBJECTIVETo understand the effect of highly active antiretroviral therapy (HAART) on AIDS patients, and to explore the prevalence and the impact of HIV-1 drug resistance in Shandong province.

METHODS2 cross-sectional studies were carried on in 2004 and 2005, to collect data on clinical symptoms and compliance of the AIDS patients with HAART through questionnaire. Informed-consent principle was followed to test on immunological, viral and laboratory index of them. HIV-1 drug genotype resistance by sequencing the gene of HIV-POL after RT-PCR was performed and analyzed.

RESULTS31 AIDS cases with and. 27 AIDS cases without HAART, were studied. 83.3% and 64.5% of the AIDS patients with HAART showed that the CD4+ T cell count was rising to over 350/microl, in the first study (2004) and in the second (2005) study respectively but still 45.8% and 45.2% of AIDS patients under HAART in the 2 years showed a decreasing HIV load under the detected limit. However, these findings were showing remarkable difference when compared with the AIDS without HAART. 7 drug resistance gene sites were found in AIDS patients with HAART and in AIDS patients without HAART. The rate on high degree drug resistance mutation and total drug resistance rate of mutation of the former were higher remarkably than those of the latter.

CONCLUSIONMost of the AIDS patients with HAART met the purpose of rebuilding immunity and control of HIV,as well as alleviation of symptoms. Although the drug resistance stain appeared in Shandong,but had little effect on HAART. AIDS; Drug resistance; Highly active antiretroviral therapy

Acquired Immunodeficiency Syndrome ; drug therapy ; Adult ; Antiretroviral Therapy, Highly Active ; CD4 Lymphocyte Count ; China ; Cross-Sectional Studies ; Drug Resistance, Viral ; genetics ; Genotype ; HIV-1 ; drug effects ; genetics ; Humans ; Middle Aged ; Patient Compliance ; Prevalence ; Surveys and Questionnaires ; Viral Load

OBJECTIVETo study the protein tyrosine phosphatase-1B (PTP1B) inhibitory activity of natural products from algae aiming at searching for new way for the treatment of type 2 diabetes mellitus (T2DM) and obesity.

METHODBromophenols derivatives from algae were screened against the PTP1B by the colorimetric assay with GST/PTP1B fusion protein. The Me2SO was distributed as the full enzyme activity, and Na3VO4 (IC50 2 micromol L(-1)) was distributed as the positive control. Inhibition rate was assayed and IC50 were calculated by LOGIT method.

RESULTThree bromophenols from Rhodomela confervoides and Leathesia nana, 3, 4-dibromo-5-(methoxymethyl)-1, 2-benzenediol (1), 2-methyl-3-(2, 3-dibromo4, 5-dihydroxy)-propylaldehyde (2) and 3-(2, 3-dibromo-4, 5-dihydroxy-phenyl)-4-bromo-5, 6-dihydroxy-1, 3-dihydroiso-benzofuran (3) showed significant inhibitory activity against PTP1B. IC50 values were 3.4 +/- micromol L(-1), 4.5 micromol L(-1) and 2.8 micromol L(-1), respectively.

CONCLUSIONThe results prove that three bromophenol derivatives from algae with significant inhibitory activity against PTP1B were potential and effective therapeutic agents for treatment of T2DM and obesity.

Diabetes Mellitus, Type 2 ; drug therapy ; metabolism ; Eukaryota ; chemistry ; Phaeophyta ; chemistry ; Phenols ; chemistry ; therapeutic use ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; antagonists & inhibitors ; Rhodophyta ; chemistry

The complete coding sequences of Eya gene family was amplified by standard PCR fromhuman tissues or cells cDNA library.The product of PCR was cloned into the eukaryotic expression vector pcDNA3-FLAG,generating pcDNA3-FLAG-Eya1～4.Thenhuman embryo kidney 293T cells were transfected with the recombinant plasmids and the expression of Eya genes were identified by Western blot.Transcriptional assay using a reporter containing myogenin enhance factor indicated that expression of Eya cooperation with Six in 293T cells affected the Myogenin gene expression.The expression vectors of Eya genes were constructed and confirmed by restriction enzyme digestion and DNA sequence analysis.Transcriptional assay using a reporter containing myogenin enhance factor indicated that expression of Eya in coordination with Six in 293T cells stimulated the Myogenin gene expression.Eya proteins are transcriptional activator of Six and can improve the activity of myogenin promoter.

Shilajit, a medicine herb commonly used in Ayurveda, has been reported to contain at least 85 minerals in ionic form that act on a variety of chemical, biological, and physical stressors. The substantia gelatinosa (SG) neurons of the trigeminal subnucleus caudalis (Vc) are involved in orofacial nociceptive processing. Shilajit has been reported to be an injury and muscular pain reliever but there have been few functional studies of the effect of Shilajit on the SG neurons of the Vc. Therefore, whole cell and gramicidin-perfotrated patch clamp studies were performed to examine the action mechanism of Shilajit on the SG neurons of Vc from mouse brainstem slices. In the whole cell patch clamp mode, Shilajit induced short-lived and repeatable inward currents under the condition of a high chloride pipette solution on all the SG neurons tested. The Shilajit-induced inward currents were concentration dependent and maintained in the presence of tetrodotoxin (TTX), a voltage gated Na+ channel blocker, CNQX, a non-NMDA glutamate receptor antagonist, and AP5, an NMDA receptor antagonist. The Shilajit-induced responses were partially suppressed by picrotoxin, a GABAA receptor antagonist, and totally blocked in the presence of strychnine, a glycine receptor antagonist, however not affected by mecamylamine hydrochloride (MCH), a nicotinic acetylcholine receptor antagonist. Under the potassium gluconate pipette solution at holding potential 0 mV, Shilajit induced repeatable outward current. These results show that Shilajit has inhibitory effects on the SG neurons of Vc through chloride ion channels by activation of the glycine receptor and GABAA receptor, indicating that Shilajit contains sedating ingredients for the central nervous system. These results also suggest that Shilajit may be a potential target for modulating orofacial pain processing.
6-Cyano-7-nitroquinoxaline-2,3-dione ; Animals ; Brain Stem ; Central Nervous System ; Chloride Channels ; Facial Pain ; Gluconates ; Mecamylamine ; Mice ; Minerals ; N-Methylaspartate ; Neurons ; Picrotoxin ; Potassium ; Receptors, Glutamate ; Receptors, Glycine ; Receptors, Nicotinic ; Resins, Plant ; Strychnine ; Substantia Gelatinosa ; Tetrodotoxin