Exosomal miR-141-3p regulates osteoblast activity to promote the osteoblastic metastasis of prostate cancer.

Exosomes from cancer cells, which contain microRNA and reach metastasis loci prior to cancer cells, stimulate the formation of a metastatic microenvironment. Previous studies have shown that exosomal miR-141-3p is associated with metastatic prostate cancer (PCa). However, the role and regulatory mechanism of miR-141-3p in the microenvironment of bone metastases require further study.

[Transcriptional activities of tumor-specific survivin promoter and PSMA promoter and enhancer in human prostate cancer: evaluation and comparison].

Objective: To detect and compare the transcriptional activities of prostate-specific membrane antigen (PSMA) promoter and enhancer and survivin promoter in different human prostate cancer cell lines, and to search for some evidence for the targeting gene therapy of human prostate cancer.

Methods: The fragments of the PSMA promoter and enhancer and survivin promoter were amplified by PCR and inserted into pGL3-Basic. The recombinant plasmids were transiently transfected into human prostate cancer cell lines and normal Chang liver cells, and, their transcriptional activities in various cells were determined by measuring the expression of luciferase.

[Effects of TFDP3 on regulating the autophagy and apoptosis of LNCaP cells].

Aim: To investigate the effect of TFDP3 on prostate cancer cell line LNCaP by transgenic method, and to explore the effect of TFDP3 on regulating the autophay and apoptosis by co-regulation with E2F1.

Methods: LNCaP cells were transfected with pcDNA3.1-TFDP3, pCMV-E2F1-HA or pcDNA3.

Evaluation of a new rapid influenza A diagnostic test for detection of pandemic (H1N1) 2009 and seasonal influenza A virus.

Background: Rapid influenza A diagnostic tests (RIDTs) play an important role in the clinical setting, especially in the influenza post-pandemic era with three influenza A viruses in circulation.

Objectives: Determine the sensitivity and specificity of a new RIDT (FluA Dot) by comparison with BD Directigen EZ FluA+B and CDC rRT-PCR.

Study Design: Two sets of experiments were conducted to determine the performance of the new test.

[Immune response induced by Rpf domain from Micrococcus luteus in mice].

Aim: To investigate the immunobiology of Rpf domain from Micrococcus luteus.

Methods: BALB/c mice were immunized with Rpf domain three times at 2-week interval. ELISA was used to detect the title of the anti-Rpf domain antibody titer in the immunized mice sera.

[Preparation and characterization of monoclonal antibodies against Micrococcus luteus Rpf domain].

Aim: To express Micrococcus luteus Rpf domain in prokaryotic cells and prepare monoclonal antibodies against Rpf domain.

Methods: The gene encoding Micrococcus luteus Rpf domain was amplified from genome of Micrococcus luteus by polymerase chain reaction(PCR), and inserted into cloning vector pUC-19. After sequenced, Micrococcus luteus Rpf domain gene was subcloned into the expression vector pPro-EXHT and transfected into E.

[Detection of transcriptional activities of tumor-specific survivin promoter in human prostatic carcinoma].

Objective: To clone DNA sequence of the survivin promoter and study is transcriptional activities in human prostate cancer cells and normal Chang liver cells.

Methods: The fragment of the survivin promoter was acquired by PCR amplification and inserted into pPRIME vectors to reconstruct a recombinant plasmid named pPRIME-S1pro and pPRIME-S2pro. Then the reconstructed plasmid was transiently transfected into human prostate cancer cells lines LNCaP and normal Chang liver cells.

[Construction of the eukaryotic expression vector of bioactive fragment of human bactericidal/permeability-increasing protein and its expression in CHO cells].

Aim: To construct the eukaryotic expression vector of the cDNA sequence encoding bioactive N-terminal fragment of human bactericidal/permeability-increasing protein (BPI) and express it in CHO cells.

Methods: Total RNA was extracted from human polymorphonuclear leukocytes (PMN) and subjected to reverse transcription, then the human BPI cDNA gene was amplified by nested PCR. The PCR product was cloned into pUC19 plasmid and confirmed by restriction enzyme digestion and DNA sequencing.

[Inhibition of survivin gene expression in PC-3 cells by RNAi].

Objective: To construct eukaryotic expression vectors by using the pSilencer3. 1-H1 neo vector for inhibiting human survivin gene by RNA interference, and to detect the effect of the silenced survivin gene on PC-3 cells.

Methods: Three target gene segments were synthesized and cloned into the pSilencer3.

Guided selection of an anti-gamma-seminoprotein human Fab for antibody directed enzyme prodrug therapy of prostate cancer.

Background: The HAMA response is a major challenge when murine antibodies are repeatedly administered for antibody directed enzyme prodrug therapy in vivo. In this study we have achieved humanization of the anti-gamma-seminoprotein E(4)B(7) murine mAb by guided selection.

Methods: Using optimal Ig Fab primers, human Fd and CL gene repertoires were amplified by RT-PCR from PBMCs of prostate cancer patients.

[Screening of human anti-gamma-seminoprotein light chain guided with the murine Fd fragment].

Aim: To screen human anti-gamma-sm (gamma-seminoprotein) light chain (Lc) with guided selection of murine Fd fragment.

Methods: The human Lc repertorie genes were amplified by RT-PCR from PBMC in patients with prostate cancer, and cloned into the phagemid vector pComb3X with murine Fd gene against gamma-seminoprotein to construct the human-mouse hybrid Fab antibody library. The size of the library, antibody gene recombinant percentage and diversity were identified by colony counting, plasmid digestion and colony sequence analysis, respectively.

[Study on the nucleic acid of E. coli bacteriophage with broad host range and its sterilization effect to sewage samples from the environment].

Objective: To study the change of nucleic acid sequence and the germicidal effect of an E. coli bacteriophage with broad host range isolated from hospital sewage as well as to study the mechanism of phage host specificity and the effect of killed bacteria by phage-disinfectant to the samples from sewage water.

Methods: To extract the nucleic acid from phage f(2) and phage with broad host range using anti-serum-carbamidine hydrochloride assay.

[The diagnostic significance of Ig heavy chain rearrangement detected in B cell lymphoma patients' serum or plasma blood samples].

Objective: To evaluate the diagnostic significance of detecting immunoglobulin (Ig) heavy chain (IgH) by using serum or plasma as blood samples.

Methods: First, collect serum and plasma blood samples of patients with B-NHL and extract tumor-derived DNAs. Then design the primer to amplify framework3 (Fr3) from the V segment regions to the J regions of the IgH complementary determining region III (CDR-III) gene.

[Detection of expression of endometriosis-related cytokine and their receptor genes by cDNA microarray technique].

Aim: To study the pathogenesis of endometriosis(EM) by investigating cytokine(CK) and CKR genes expression involved in the development of EM.

Methods: The CK and CKR gene expression pattern in samples of EM and normal endometrial tissues were analyzed by using cDNA microarrays.

Results: 119 genes were expressed differently between 3 cases EM tissues and 3 cases normal endometria tissues.

[Construction of antisense recombinant adenoviral vector for c-myc and its antiproliferative effect on rat lymphocytes].

Aim: To observe the antiproliferative effect of antisense recombinant adenoviral vector for c-myc on rat thymus lymphocytes.

Methods: Antisense and sense bacterial plasmids for c-myc were constructed. Bacterial plasmids and El detected adenoviral plasmid were cotransfected into 293 cells.

[Study on isoniazid-resistant Mycobacterium tuberculosis isolates by multiple-polymerase chain reaction-single strand conformation polymorphism].

Objective: To develop a new multiple-polymerase chain reaction-single strand conformation polymorphism (multi-PCR-SSCP) system for detecting the aphC promoter, inhA, and katG gene mutations in isoniazid-resistant Mycobacterium tuberculosis isolates in the single reaction, and for the quick diagnosis of isoniazid-resistant Mycobacterium tuberculosis isolates.

Methods: Three pairs of oligonucleotide primers were designed according to the aphC promoter, inhA, and katG genes of Mycobacterium tuberculosis to examine isoniazid-resistance by multi-PCR-SSCP.

Results: Isoniazid-sensitivity and resistance were analyzed with general PCR and multi-PCR at the same time, and H(37) Rv was used as a control.