Thyroid hormone is an important hormone for regulating cell differentiation,growth and metabolism.Many studies have shown that thyroid hormone can induce turmor angiogenesis after binding with the integrin ανβ3 receptor at cell surface.Tetraiodothyroacetic acid is the analogue of thyroid hormone,and can inhibit tumor angiogenesis by inhibiting the binding of thyroid hormone with integrin ανβ3.Studying the role of thyroid hormone in promoting tumor angiogenesis may provide a new approach for molecular targeted therapy of tumor.

Adult ; Aged ; Cervical Intraepithelial Neoplasia ; pathology ; surgery ; Cervix Uteri ; pathology ; surgery ; Conization ; methods ; Female ; Follow-Up Studies ; Humans ; Hysterectomy ; methods ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm, Residual ; pathology ; surgery ; Uterine Cervical Neoplasms ; pathology ; surgery

Adult ; Carcinoma, Endometrioid ; diagnosis ; pathology ; Cervix Uteri ; pathology ; Diagnosis, Differential ; Endometrial Neoplasms ; diagnosis ; pathology ; Endometrial Stromal Tumors ; diagnosis ; pathology ; Female ; Humans ; Middle Aged

Objective To characterize the dimerization and the antigenicity of the ORF2 polypep-tide of hepatitis E virus (HEV, genotype 4). Methods HEV ORF2 gene was cloned from the serum of a patient with hepatitis E. The genotype was determined by sequencing. Three ORF2 polypeptides differing in size and other polypeptides with point mutations were produced in E. coli. The recombinant polypeptides were purified and analyzed by SDS-PAGE and Western blot. Results The ORF2 polypeptide containing 459-607 amino acid formed homedimer even in 8 mol/L urea. The truncated polypeptides containing amino acid 472-607 or 459-594 formed monomer only. The mutations at amino acid 562 or 595 disrupted the ho-modimer, whereas the mutations at amino acid 476 or 580 did not. Anti-HEV from hepatitis E patients only reacted with the homodimer form of the polypeptide 459-607 and did not react with monomer or tnmcated pol-ypeptides. Conclusion The amino acid 459-607 of HEV ORF2 is essential for dimerization of the ORF2 polypeptide. Residues at amino acid 562 and 595 are critical for the dimerization. The antigenicity of the polypeptide 459-607 mainly depends on its homodimer form.

Objective To elucidate intracellular transcription factor activation of C_7～T_5 dorsal root ganglia in rats recurrently infected with respiratory syncytial virus (RSV). Methods Eighty 1～2 weeks old Sprague-Dawley rats were randomly divided into 2 groups: a control group and a RSV-infection group. The rats in the RSV-infection group were infected with 5 ×10~5 U/mL RSV once a week and the rats in the control group were treated with culture medium without RSV. Airway response was measured after 8 weeks. Lung tissue was submitted for HE staining and in situ hybridization. The C_7～T_5 dorsal root ganglia were obtained for the preliminary screening of the intracellular transcription factors by TranSignal~(TM) protein/DNA combo array. Nuclear protein of C_7～T_5 dorsal root ganglia were extracted and submitted to Western blot. Results Airway response in the RSV-infection group was higher than that in the control group (P＜0.05). HE staining showed inflammatory cell infiltration, and in situ hybridization demonstrated positive RSV RNA in the RSV-infection rat lung which was not present in the control group, thus validating the efficacy of our model. TranSignalTM protein/DNA combo array screening showed that 55 transcription factors increased by at least 2 folds in the C_7～T_5 DRG cells of the RSV-infection group. The transcription factors Smad and interferon regulatory factor (1 or 2) were the 2 most upregulated transcription factors identified by combo array screening (59 and 43 fold increase compared with the control, respectively). Western blot confirmed Smad(1/2/3) and IRF-1 upregulate while IRF-2 remained unchanged. Conclusion Respiratory syncytial virus infection results in airway hyperresponsiveness and transcription factor activation in C7～T5 spinal adorsal root ganglia in rats, which may contribute to airway nerve network dysfunction and airway hyperresponsiveness.

Objective To exclude false positivities in detection of IgM antibodies against hepatitis E vires of genotype 4 (HEV-4) using a truncated immunodominant polypeptide of HEV open reading frames (ORF2). Methods The recombinant ORF2 immunodominant polypeptide corresponding to amino acids (AA) 459-607 and a truncated polypeptide corresponding to AA 472-607 were separately applied to coat ELISA plates. Anti-HEV IgM from 35 serum samples with HEV RNA positive, 69 serum samples from healthy individuals and 117 clinically suspicious HEV RNA positive serum samples was detected by an indirect ELISA and was confirmed by western blot in protein level and RT-PCR detecting in RNA level. Results Western blot analysis showed that the sera from HEV patients reacted with the dimmer of peptide 459-607, but they didn't react with the monomer and peptide 472-607. The ELISA showed that all 35 serum HEV RNA positive samples reacted with peptide 459-607 but not with peptide 472-607 and none of the 69 serum samples from healthy individuals reacted with either polypeptide. Among 117 chnically suspicious HEV RNA serum samples, 5 samples reacted simultaneously with both polypeptides. But the difference between 450 nm absorbance (A450) value was less than 0. 5. Western blot analysis demonstrated that all the 5 serum samples were anti-HEV IgM- negative. The 5 serum samples was detected negative by RT-PCR, indicating that the false pesitivities were caused by non-specific absorption. Conclusions ORF2 peptide 459-607 may be used to detect anti-HEV lgM efficiently. The false positivities caused by non-specific absorption can be largely excluded according to the difference between 45Ohm absorbance (A450) value when serum reacts with both polypeptides.

Objective To compare the biological characteristics of several different anxiety rat models established by different methods of stress at different time points and provide experimental basis for the most appropriate modeling methods .Methods 60 rats were randomly divided into normal , empty bottle stress , chronic emotional stress ( CES ) group, restraint stress for 3h, 6h, and modeling respectively .In the experimental 7 d, 14 d, 21 d, elevated plus maze and fear condition system was used to test anxiety-like behavior in rats , open field test to study anxiety or depression-like behavior , forced swimming test was used to detect depression-like behavior in rats , and using the Elisa test kit to detect the contents of 5-HT, DA in the hippocampus in rats .Results Anxiety-like behavioral test results showed that rats in empty bottle stress, CES, 6 h restraint stress group started to have anxiety-like behavior since 14 d, then anxiety-like behavior was becoming increasingly apparent .Forced swimming test results showed that immobility time in 6 h restraint rats was significantly increased in the first 7 d(P <0.05).Meanwhile, compared with control group, hippocampal 5-HT, DA contents in empty bottle stress and CES rats increased significantly since 14 d.Conclusions Among several stress methods established anxiety model , anxiety-like behavior in 3 h restraint stress was not obvious; 6 h restraint stress exhibited a depression-like behavior in the forced swimming test might be due to prolonged stress .Empty bottle stress and CES can successfully establish the anxiety rat model , and the anxiety behavior of the rats have some differences . Corresponding model methods can be selected according to different experimental purposes .

Objective To evaluate the follow-up results of patients with cervical intraepithelial neoplasia Ⅲ(CIN Ⅲ)after surgical treatment.Methods A retrospective analysis of consecutive patients with CIN Ⅲ after surgical treatment between Jan 1st,1999 and Jun 30 th,2004 was performed.The follow- ups of the patients after surgical treatment were assessed.Results In the follow-up of patients with CINⅢ after surgical treatment,the rate of abnormal cytology was 9.3%.The rate of follow-up was higher in patients with cervical conization than in patients with initial hysterectomy and in patients of the oncological group than of the non-oncological group.The rate of follow-up was lower in patients over 40 years old.There was no difference in the residential areas of the patients.Conclusions Cytological follow-up of patients with CINⅢ after operation is varied.The rate of follow-up is lower in patients over 40 years old,in patients having initial hysterectomy and in patients of the non-oncological group.The rate of follow-up is associated with the knowledge about CIN of both surgeons and patients.

Aims To study the effects of clenbuterol on anoxia/reoxygenation( A/R) injury in neonatal Wistar rat cardiomyocytes and to explore whether its mecha-nism is related to reperfusion injury salvage kinase ( RISK) or not. Methods The cultured primary neo-natal cardiomyocytes were randomly divided into eight groups: ①normal culture group; ②anoxia/reoxygen-ation( A/R) group;③ clenbuterol ( 1 μmol · L-1 ) +A/R;④ICI118,551(10 μmol·L-1) + clenbuterol ( 1 μmol · L-1 ) + A/R; ⑤Metoprolol ( 10μmol · L-1 ) + clenbuterol(1μmol·L-1 ) + A/R group;⑥Metoprolol ( 10 μmol · L-1 ) + A/R group; ⑦PD98059 ( 20 μmol · L-1 ) + clenbuterol ( 1 μmol · L-1 ) + A/R group;⑧ LY294002(10 μmol·L-1 ) +clenbuterol(1 μmol · L-1 ) + A/R group. Cell via-bility was determined by the conventional MTT reduc-tion assay. The content of LDH in cultured medium was measured with colorimetry. Cardiomyocyte apopto-sis was determined by Hoechst33342 . Intracellular re-active species( ROS) were monitored by the fluorescent DCFH-DA. Total ERK2 and phosphorylated ERK were detected by western blot. Results Compared with A/R group, clenbuterol significantly increased vaibility of cells, reduced LDH release, lowered the rate of apop-tosis and ROS production. When addedβ2 receptor an-tagonist ICI118 , 551 , PI3 K inhibitor LY294002 and ERK inhibitor PD98059 , the effects of clenbuterol a-bove were inhibited; but β1 receptor antagonist Meto-prolol protected the cardiomyocytes from A/R injury, as evidenced by decreased LDH release and increased cell viability. There were no synergistic effects in the combined use of clenbuterol and Metoprolol. Conclu-sion clenbuterol exerts cardioprotective effects against A/R injury by inhibiting oxidative stress and apopto-sis. The protection of clenbuterol is inhibited by ICI118 , 551 , LY294002 and PD98059 . clenbuterol protects cardiomyocytes against A/R injury via RISK pathway by activation of β2 receptor.

Objective To investigate the regulating effects of Zuogui Jiangtang Jieyu Formula (ZJJF) on Bcl-2, Bax and Caspase-8 in hippocampus neuron damage in diabetes mellitus with depression (DD). Methods Hippocampal neurons from 18 d pregnant rats were primitively cultivated, and then combination of glucose and corticosterone was used to construct DD simulation environment. Cultivated hippocampal neurons were randomly divided into normal group, blank serum group, model group, positive medicine (metformin+fluoxetine) serum group and to-be-tested medicine (ZJJF) serum group. Normal group and model group were given same amount culture medium, while other group were given relevant amont of 10％ medicine serum or blank serum. After modeling intervention for 18 h,Hoechst staining was used to detect the apoptosis of hippocampal neurons. The expression of Bcl-2, Bax and Caspase-8 was detected by high content analysis. Results Compared with the control group, hippocampal neuron dendrites ruptured or decreased, neural network connection decreased, cells showed significant staining, broken, uneven distribution of light spots, the expression of Bcl-2 protein decreased significantly (P<0.05), but Bax and Caspase-8 were increased (P<0.05, P<0.01). Compared with the model group, hippocampal neurons in both positive medicine serum group and ZJJF serum group gradually recovered. Hoechst staining showed that the nuclei were significantly homogenized, local highlights were significantly reduced, Bcl-2 protein expression levels were significantly increased (P<0.05, P<0.01), while Bax and Caspase-8 were obviously down-regulated (P<0.05, P<0.01). Conclusion ZJJF has protective effects on hippocampal neurons in DD of model rats, and its mechanism is related to regulating the expression of Bcl-2, Bax and Caspase-8 in hippocampus neuron.