A serum inhibited gene Si1(GenBank acession number:AY050169) was previously cloned and identified by differential expression of genes in U251 cells.For the further study of biological function of Si1, prediction procedure was performed to predict its subcelluar-localization.Relative experiments were carried out at the same time.The expression of EGFP/Si1 recombinant in HeLa cells showed Si1 protein located in nuclear which corroborated the prediction results of PsortⅡ, Proloc, Cello version2, Subnuclear compartments prediction system, NUCLEO and NUCPRED.According to the PredictNLS prediction, twelve different fragments of EGFP/Si1 recombinants were constructed to identify precise NLS regulation sequence.Findings proved that the real NLS regulation sequence was not the same as the software predicted(1 206 bp～1 239 bp on Si1 ORF), but located on 1 395 bp～1 594 bp of Si1.A tumor relatived mutation/EGFP recombinant localization result showed though the mutation site(1 639bp on Si1 ORF) does not located in NLS regulation sequence, it did affect wildtype Si1 protein divert to nuclear and may affect its natural function in cell, perhaps it is the main reason for highly mutation rate of Si1 in tumor.

Objective To study on the relations between endometrial carcinoma and Infections of HSVⅡ.Methods Using to skill of PCR-SSCP to test 30 cases of endometrial carcinoma,others 15 cases of the hyperplastic endometrial and 15 cases of the normal of endometrial were infected HSVⅡ.Results Positive rates of endometrial carcinoma that was infected of HSVⅡ were 50%(15/30),Positive rates of the hyperplastic endometrial that was infected of HSVⅡ were 6.67%(1/15),15 cases of the normal of endometrials were not infected.Compared with the endometrial carcinoma and the hyperplastic endometrial and the normal of endometrial,P

Objective To evaluate the effect of argon laser photocoagulation (photocoagulation group) and endolaser photocoagulation during vitrectomy (vitrectomy group) on the eyes with Eales disease by fundus fluorescein angiography(FFA) and visual acuity (VA).Methods Sixty-seven eyes with Eales were treated by photocoagulation and 31 eyes with Eales were treated by vitrectomy. The changes of FFA and VA before and after treatment were observed.Results In the photocoagulation group, retinal non-perfusion disappeared in 31eyes(55 4%),alleviated in 17eyes(30 4%),unchanged in four eyes(7 1%)and deteriorated in four eyes(7 1%).Retinal neovascularization disappeared in 30 eyes(50 8%),alleviated in 19 eyes (32 2%),remained in six eyes(10 2%)and deteriorated in four eyes(6 8%).The first FFA after treatment, non-perfusion was existed in 25 eyes in photocoagulation group, and only was existed in two eyes in vitrectomy group.Neovascularization was existed in 29 eyes in photocoagulation group, and was existed in three eyes in vitrectomy group.The best visual correction(VA≥3 0) in photocoagulation and vitrectomy groups respectively was 30 eyes and zero eye before treatment,and after treatment respectively was 47 eyes and five eyes.There were 54 eyes and 11 eyes with VA≥0 05 in photocoagulation and vitrectomy groups respectively before treatment,and 60 eyes and 20 eyes with VA≥0 5 respectively after treatment.Conclusions The better result of FFA in vitrectomy group was related to the sufficient photocoagulation by endolaser. The higher rate of low vision in vitrectomy group was related to the impairment of macula by large amount of vitreous hemorrhages,retina detachment and vitrectomy. So the patients with Eales should be checked by FFA as early as possible.If the patients had the indicator to be treated by laser,they should be treated early and rechecked by FFA,to avoid the visual function harm by repeated vitreous hemorrhage due to incomplete treatment.

Objective:To construct a phage display antibody library of special antigen responding to serum starved U251 cell,from which the serum responding gene and protein would be gotten.Methods:U251 cells were cultured in serum-absent midium for 48 h.Its protein was extracted and used to immunize BALB/c mice.Total RNA of the spleenocytes of immunized mice was extracted.VH and VL were amplified by RT-PCR and were linked into ScFv(Single chain fragment of variation) with a linker.ScFv was recombined to pCANTAB5E vector and was transformed to TG1 strain.Results:The library capacity was up to 3?106 cfu/L.A positive clone was identified from 8 random clones of this library.Conclusion:The special ScFv phage library is constructed successfully.It is the basis for screening special antibodies which can recognize serum responding protein.

Objective To observe the effect of Xiaoxianxiong Decoction combined with metformin on glucose and lipid metabolism of phlegm-heat type 2 diabetic patients (T2DM). Methods Totally 104 phlegm-heat T2DM patients were randomly divided into two groups (each contains 52 cases), and given the diabetes lifestyle intervention. Control group additionally took 0.5 g metformin twice daily. And treatment group, besides taking 0.5 g metformin, was given Xiaoxianxiong Decoction, one dose and three times per day warmly. After 12 weeks, FBG, HbA1C, TC, TG, HDL-C, LDL-C, 2 hPG of the two groups were determined, and the therapeutic effect was evaluated. Results The total effective rate in the treatment group was 92.31%, and 76.92%in control group (P<0.01). The difference in symptom score, FBG, 2 hPG, TC, TG, HDL-C and LDL-C between the two group were significant, so as that between before and after treatment of both groups (P<0.05). Conclusion Xiaoxianxiong Decoction combined with metformin can effectively lower the levels of blood glucose and blood lipid in phlegm-heat T2DM patients, enhance the therapeutic effect and improve the metabolism of glucose and lipid.

Human arrest defective 1 (hARD1) is an acetyltransferase; its physiological significance remains unclear. To explore the relationship between ARD1 protein and tumors, we detected the hARD1 protein in tumor tissues in vivo. We cloned hARD1 gene from Hela cell and construct recombinant plasmid pET28b-hARD1. The recombinant plasmid was transformed into E. coli BL21 (DE3)plysS. hARD1 protein was expressed by inducing with IPTG(1 mmol/L) and purified up to 95% through Ni2+ chelation affinity chromatography. We used the purified hARD1 protein as antigen immunized the Balb/c mice and obtained the hARD1 specific polyclonal antiserum. Through immunohistochemical analysis of different tumor tissues in vivo, we found that hARD1 expressed at high frequency in breast cancer, prostate cancer and lung cancer, especially, hARD1 expression frequency in breast cancer was up to 70%, which is higher than in the other tumors. These results indicate that the high expression level of hARD1 could be an indicator of the breast cancer. This new finding would be a foundation to further explore the relationship between breast tumor and hARD1.
Acetyltransferases ; analysis ; genetics ; immunology ; Amino Acid Sequence ; Animals ; Antibodies ; blood ; immunology ; Base Sequence ; Biomarkers, Tumor ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Female ; Humans ; Immune Sera ; biosynthesis ; Immunization ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; N-Terminal Acetyltransferase A ; N-Terminal Acetyltransferase E ; Prostatic Neoplasms ; metabolism ; pathology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology

The loci of cDNA sequences for valid diagnosis have been identified through the selection of the genome of SARS coronaries. The gene chips for diagnosing such virus have been developed, based on our own-developed technology for manufacturing and application of gene chips. The diagnoses given by such gene chips were consistent well with the reports of clinical laboratories (94.29%) and the sensitivity reached 10(-2)/ml virus molecules. This method is well suited for the clinical use in SARS coronaries diagnosis.
Humans ; Oligonucleotide Array Sequence Analysis ; SARS Virus ; isolation & purification ; Sensitivity and Specificity ; Severe Acute Respiratory Syndrome ; virology

Human arrest defective 1(hARD1) is an acetyltransferase catalyzing the N-terminal acetylation of proteins after translation. The high expression of hARD1 could be an indicator of the breast cancer. In current study, we produced an anti-hARD lp monoclonal antibody that could specifically recognize ARD1 in breast cancer tissues by using the immunohistochemical assay. The full-length His-tag hARD1 protein (1-235 aa) was over-expressed in Escherichia coli, and purified recombinant protein was injected into Balb/c mice to perform immunization procedure. Eight stable positive monoclonal cell lines were isolated. ELISA results demonstrated that all light chains of antibodies were kappa, and the heavy chains displayed three subtypes IgG1, IgG2a and IgG2b, respectively. A monoclonal antibody, which could specifically recognize hARD1 protein in breast cancer tissues, was identified by screening different cancer tissues using antibody-specificity method. Further, the specificity of the antibody was confirmed by Western blotting analysis. Our study would facilitate breast cancer diagnosis by using this ARD1 monoclonal antibody in clinic. Also, this antibody could be used as an important tool for further investigating the role of ARD1 in tumorigenesis.
Acetyltransferases ; genetics ; immunology ; Animals ; Antibodies, Monoclonal ; biosynthesis ; genetics ; immunology ; Biomarkers, Tumor ; biosynthesis ; immunology ; Breast Neoplasms ; immunology ; metabolism ; pathology ; Escherichia coli ; genetics ; metabolism ; Female ; Humans ; Immunization ; Mice ; Mice, Inbred BALB C ; N-Terminal Acetyltransferase A ; N-Terminal Acetyltransferase E ; Recombinant Proteins ; biosynthesis ; genetics ; immunology