Objective To investigate the signal transduction of bFGF to regulate the colorectal carci- noma cell growth via PI3K/PKB pathway.Methods MTT assay was used to determine the suppressive effect of bFGF on the growth rate of LoVo cell;(?-~(32)P)ATP incorporation assay was used to detect the increased activity of PKB in LoVo cell;the RT-PCR technique was used to test the expression of cyclin A;Western Blot analysis was used to detect the expressions of PKB and cyclin A protein.Results When the bFGF worked on the LoVo cell at various times,it was found that the PKB activity of the LoVo cell was increased by the(?-~(32)P) ATP incorporation assay.By preprocessing by the depressor-LY294002 and bFGF,the activity of PKB was sig- nificantly reduced(P

OBJECTIVETo investigate effects of zearalenone (ZEA) on the proliferation of SK-N-SH human neuroblastoma cells in vitro and its possible mechanism.

METHODSSK-N-SH cells were cultured in estrogen-free improved minimum essential medium and divided into 5 groups based on different treatments: group 1, without treatment; group 2, treated with 17beta-estradiol (E(2)); group 3, treated with ZEA; group 4, treated with both E(2) and ICI 182780; group 5, treated with both ZEA and ICI 182780. Absorbance value (AV) was determined at the time point of 0, 24, 48 and 72 hours, and DNA proliferation index (PI) at 72 hours. Flow cytometer, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) were employed to monitor cell apoptosis.

RESULTSAt 24, 48 and 72 hours, the AV of group 3 were 1.39, 1.32, and 1.22 times to those of group 1, respectively. PI in group 3 was 1.43 times of that in group 1 at 72 hours. The results of group 2 were similar to those in group 3. At the same time, the growth of cells was inhibited by ICI 182780 despite the presence of E(2) and ZEA. Apoptosis cells were abundant in group 1 and ICI 182780 groups, but little in E(2) and ZEA groups.

CONCLUSIONZEA might promote the proliferation of SK-N-SH cells to a level similar to that of E(2), which might probably be brought about via estrogen receptor pathways and depressing apoptosis.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Neuroblastoma ; Receptors, Estrogen ; antagonists & inhibitors ; Zearalenone ; toxicity

To investigate the effect of Jiawei Foshou San and its various combined administration on hepatic P450 enzyme activity and hepatocyte morphology in rats. Rats were orally administered with drugs for four weeks and then sacrificed to prepare liver microsomes. The liver microsomes were incubated with the cocktail method; The metabolites were determined with the rapid liquid chromatography with tandem mass spectrometry (LC-MS/MS) to investigate the hepatocyte P450 enzyme activity. In addition, the hepatic pathological changes were observed by using the hematoxylin and eosin (HE) staining. Compared with the control group, the enzyme activity of CYP1A2 and CYP3A4 in the Jiawei Foshou san group showed a significant rise (P < 0.05); the enzyme activity of CYP1A2, CYP2C9, CYP2D6, CYP2E1 and CYP3A4 in the ferulic acid + ligustrazine group and the ligustrazine + tetrahydropalmatine group showed a significant rise (P < 0.05) ; the enzyme activity of CYP1A2, CYP2D6 and CYP2E1 in the ligustrazine group showed a significant rise (P < 0.05); the enzyme activity of CYP3A4 in the ferulic acid group showed a significant reduction (P < 0.05). After the administration with various drugs, the hepatocyte morphologies in the ferulic acid group and the ligustrazine group were normal. The pathological changes were observed in the tetrahydropalmatine group, such as unclear boundary of hepatic lobules, disordered hepatic cell arrangement, blurred edge, anisokaryosis and infiltration of inflammatory cells. The ferulic acid + tetrahydropalmatine group, the ligustrazine + tetrahydropalmatine group and the Jiawei Foshou San group also showed inflammatory infiltration, but with less pathological changes, particularly the Jiawei Foshou San group. The study result shows that Jiawei Foshou San can induce the enzyme activity of CYP1A2 and CYP3A4, and ligustrazine may be the effective substance for inducing CYP1A2. Its combination with ferulic acid and ligustrazine can significantly reduce the liver toxicity of tetrahydropalmatine.
Animals ; Cytochrome P-450 Enzyme System ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatocytes ; drug effects ; enzymology ; Microsomes, Liver ; drug effects ; enzymology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of lymphadenectomy adjacent to inferior mesenteric artery root on the prognosis of rectal cancer.

METHODSClinicopathological data of 260 cases with rectal cancer undergone radical operation were analyzed retrospectively. The patients were divided into two groups. Group D(2): the lymph nodes adjacent to mesenteric artery root were not excised (n=188). Group D(3): the lymph nodes adjacent to mesenteric artery root were excised (n=72). Prognosis of two groups was compared during the follow-up period.

RESULTSIn group D(2), the 1-, 3-, 5-year total survival rates (TS) were 97.3%, 87.2% and 77.1%, and tumor-free survival rates (TFS) were 93.1%, 83.0% and 76.8% respectively. In group D(3 ), the 1-, 3-, 5-year total survival rates (TS) were 94.4%, 79.2% and 73.6%, and tumor-free survival rates (TFS) were 86.1%, 76.4% and 71.0% respectively. The differences of TS and TFS between two groups were not significant according to Kaplan-Meier analysis (P>0.05). Multivariate analysis revealed that the excision of lymph nodes adjacent to mesenteric artery root was not statistically correlated with the recurrence, metastasis and survival time after radical operation of rectal cancer.

CONCLUSIONExcision of lymph nodes adjacent to inferior mesenteric artery root has no significant impact on prognosis and it is unnecessary in the radical operation of rectal cancer.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lymph Node Excision ; methods ; mortality ; Lymph Nodes ; surgery ; Lymphatic Metastasis ; Male ; Mesenteric Artery, Inferior ; surgery ; Middle Aged ; Prognosis ; Rectal Neoplasms ; mortality ; pathology ; surgery ; Survival Rate ; Treatment Outcome

OBJECTIVETo investigate the role of the TEKT4 protein in the pathogenesis of idiopathic asthenozoospermia.

METHODSWe separated and purified the ejaculated sperm from idiopathic asthenozoospermia patients and normozoospermic men by Percoll discontinuous density gradients, and detected the distribution and the expressions of TEKT4 mRNA and TEKT4 protein by RT-PCR and Western blot.

RESULTSRT-PCR revealed that the expression of TEKT4 mRNA was significantly lower in the sperm of the idiopathic asthenozoospermia patients than in those of the normozoospermic men (0.59 +/- 0.13 vs 0.75 +/- 0.15, t = 4.325, P < 0.05), and Western blot confirmed the results of RT-PCR (0.48 +/- 0.14 vs 0.69 +/- 0.13, t = 5.939, P < 0.05).

CONCLUSIONThe expression of TEKT4 is significantly decreased in the ejaculated sperm of idiopathic asthenozoospermia patients, which might be one of the causes of idiopathic asthenozoospermia.

Adult ; Asthenozoospermia ; metabolism ; Blotting, Western ; Case-Control Studies ; Cytoskeletal Proteins ; metabolism ; Humans ; Male ; RNA, Messenger ; genetics ; Sperm Motility ; Spermatozoa ; metabolism

OBJECTIVETo investigate the role of the SEPT4 protein in the pathogenesis of idiopathic asthenozoospermia.

METHODSSamples of ejaculated sperm from idiopathic asthenozoospermia patients and normozoospermic men were separated and purified by Percoll discontinuous density gradients, the distribution and expression of SEPT4 in the sperm samples were determined by immunocytochemistry, and the expressions of SEPT4 mRNA and SEPT4 protein were detected by RT-PCR and Western blot.

RESULTSImmunocytochemistry showed that the expression of SEPT4, located in the annulus, was significantly reduced in the sperm of the idiopathic asthenozoospermia patients (t = 3.452, P < 0.01). RT-PCR revealed that the expression of SEPT4 mRNA was significantly lower in the sperm of the idiopathic asthenozoospermia patients than in those of the normozoospermic men (t = 3.521, P < 0.05). Western blot confirmed the results of RT-PCR (t = 5.872, P < 0.05).

CONCLUSIONThe expression of SEPT4 is significantly decreased in the ejaculated sperm of idiopathic asthenozoospermia patients, which might be one of the causes of idiopathic asthenozoospermia.

Adult ; Asthenozoospermia ; metabolism ; Case-Control Studies ; Humans ; Male ; Septins ; metabolism ; Sperm Motility ; Spermatozoa ; metabolism ; Young Adult

OBJECTIVETo investigate the changes of serum reproductive hormones with male aging and to compare the differences in the hormone levels among different age groups or between township and rural males of the same age group.

METHODSUsing cluster and stratified sampling, we recruited 434 healthy old and middle-aged (40-69 years) males, 198 from the township and 236 from the rural communities. We determined the concentrations of serum total testosterone (tT), luteinizing hormone (LH) and sex hormone binding globulin (SHBG), free testosterone (fT), bio-available testosterone (Bio-T), and obtained the testosterone secretion index (TSI) and free testosterone index (fTI). Meanwhile, we included fifty-nine 20-39 years old males from the same communities in a control group.

RESULTSWith the increase of age, the serum tT levels did not change significantly, while the levels of serum LH and SHBG increased, and those of fT, Bio-T, TSI and fTI decreased gradually. Statistically significant differences were found among the four different age groups in all the parameters of reproductive hormones (P < 0.01), except in the serum tT level (P > 0.05). The serum tT level was not significantly correlated with aging and LH (P > 0.05). Serum LH and SHBG had a marked positive correlation with aging, and SHBG with LH (P < 0.01), while fT, Bio-T, TSI and fTI were negatively correlated with aging and the LH level (P < 0.01). Serum LH, TSI and fTI showed statistical differences (P < 0.05), while fT and Bio-T exhibited extremely significant differences (P < 0.01) between the township and rural males in the 40 -49 yr group, and in the same age group, the increase rates of serum LH and SHBG and reduction rates of fT, Bio-T, TSI and fTI were higher in the rural men than in the township residents. However, the results were just the opposite in the 50 - 59 and 60 - 69 yr groups.

CONCLUSIONThe levels of serum LH, SHBG, fT, Bio-T, TSI and fTI changed with aging in a gradientmanner in the old and middle-aged males, but no significant changes were observed in the level of serum tT. There were statistical differences in many parameters of serum reproductive hormones among different age groups or between township and rural males.

Adult ; Age Factors ; Aged ; Aging ; metabolism ; China ; Humans ; Luteinizing Hormone ; blood ; Male ; Middle Aged ; Rural Population ; Serum Albumin ; metabolism ; Sex Hormone-Binding Globulin ; metabolism ; Testosterone ; blood ; Urban Population ; Young Adult

Adolescent ; Adult ; China ; epidemiology ; Female ; Humans ; Hypertension ; epidemiology ; Male ; Middle Aged ; Prevalence ; Urban Population ; Young Adult

A GeXP based multiplex PCR assay was developed to simultaneously detect six different chicken respiratory viruses including H5, H7, H9 subtypes of avian influenza virus(AIV), new castle disease virus (NDV), infectious bronchitis virus(IBV) and infectious laryngotracheitis virus(ILTV). According to the conserved sequences of genes of each pathogen, seven pairs of specific primers were designed, and the reaction conditions were optimized. The specificity and accuracy of GeXP were examined using samples of single and mixed infections of virus. The sensitivity was evaluated by performing the assay on serial 10-fold dilutions of cloned plasmids. To further evaluate the reliability, thirty-four clinical samples were detected by GeXP. The corresponding specific fragments of genes were amplified. The detection limit of GeXP was 10(2) copies/microL when all of 7 pre-mixed plasmids containing target genes of six chicken respiratory viruses were present. In the detection of thirty-four clinical samples, the results of GeXP were accorded with the viral isolation completely. In conclusion, this GeXP assay is a rapid, specific, sensitive and high-throughput method for the detection of chicken respiratory virus infections. It can be applied in rapid differential diagnosis for clinical samples, and also provide an effective tool to prevent and control chicken respiratory diseases with similar clinical symptoms.
Animals ; Chickens ; Influenza A virus ; classification ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; diagnosis ; virology ; Multiplex Polymerase Chain Reaction ; methods ; Poultry Diseases ; diagnosis ; virology ; Respiratory Tract Infections ; diagnosis ; veterinary ; virology

This study was aimed to investigate the effect of homoharringtonine (HHT) on K562 cell proliferation, apoptosis and expression of BCL-2 and NF-κB proteins. The cells proliferation was assayed with MTT method, the cell apoptosis, cell cycle and BCL-2 expression were analyzed with flow cytometry, NF-κB protein expression was detected with Western blot. The results showed that HHT concentration-dependently inhibited proliferation of K562 cells, the IC50 at 48 h was 43.89 ng/ml. Treated with HHT 10 ng/ml for 48 h, K562 cell apoptosis significantly increased, cell cycle was blocked at G0/G1, the expression level of BCL-2 and NF-κB proteins was lower than that in control group (P < 0.05). It is concluded that HHT may inhibit the proliferation of K562 cells, and down-regulating expression levels of BCL-2 and NF-κB may be one of its anti-CML mechanisms.
Flow Cytometry ; Harringtonines ; pharmacology ; Humans ; K562 Cells ; NF-kappa B ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism