YAP-galectin-3 signaling mediates endothelial dysfunction in angiotensin II-induced hypertension in mice.

Background: Vascular endothelial dysfunction is regarded as an early event of hypertension. Galectin-3 (Gal-3) is known to participate in various pathological processes. Whilst previous studies showed that inhibition of Gal-3 effectively ameliorates angiotensin II (Ang II)-induced atherosclerosis or hypertension, it remains unclear whether Ang II regulates Gal-3 expression and actions in vascular endothelium.

AMPK upregulates K2.3 channels and ameliorates endothelial dysfunction in diet-induced obese mice.

The opening of endothelial small-conductance calcium-activated potassium channels (K2.3) is essential for endothelium-dependent hyperpolarization (EDH), which predominantly occurs in small resistance arteries. Adenosine monophosphate-activated protein kinase (AMPK), an important metabolic regulator, has been implicated in regulating endothelial nitric oxide synthase activity.

Gal-3 (Galectin-3) and K3.1 Mediate Heterogeneous Cell Coupling and Myocardial Fibrogenesis Driven by βAR (β-Adrenoceptor) Activation.

Heart failure is associated with sympatho-βAR (β-adrenoceptor) activation and cardiac fibrosis. Gal-3 (galectin-3) and K3.1 channels that are upregulated in diverse cells of diseased heart are implicated in mediating myocardial inflammation and fibrosis.

K3.1 channel mediates inflammatory signaling of pancreatic β cells and progression of type 2 diabetes mellitus.

Chronic islet inflammation is associated with development of type 2 diabetes mellitus (T2DM). Intermediate-conductance calcium-activated K (K3.1) channel plays an important role in inflammatory diseases.

K3.1 Channels Promote Cardiac Fibrosis Through Mediating Inflammation and Differentiation of Monocytes Into Myofibroblasts in Angiotensin II -Treated Rats.

Background Cardiac fibrosis is a core pathological process associated with heart failure. The recruitment and differentiation of primitive fibroblast precursor cells of bone marrow origin play a critical role in pathological interstitial cardiac fibrosis. The K3.

[Preparation and identification of type specific and conformation dependent HPV16 L1 protein monoclonal antibody.].

Aim: To generate type specific and conformation dependent monoclonal antibodies against human papillomavirus 16 major capsid L1 protein (HPV16 L1).

Methods: HPV16 L1 protein was expressed by modified insect-baculovirus expression system and purified by ProBond(TM); purification system in nature conditions. BALB/c mice were immunized with the purified HPV16 L1 protein.

[Clinicopathological significance of cytotoxic lymphocytes in breast cancer and draining lymph nodes].

Objective: To analyze retrospectively the quantity and activation status of the tumor infiltrating cytotoxic lymphocytes in breast cancer and the draining lymph nodes, and its relation to the clinical pathological significance.

Methods: Seventy-four breast cancer samples with their corresponding axillary lymph nodes were histologically typed and staged. Cytotxic lymphocytes were analyzed by immunohistochemistry with the monoclonal antibodies against CD8, CD56, granzyme B and perforin.

Anti-CD3 scFv-B7.1 fusion protein expressed on the surface of HeLa cells provokes potent T-lymphocyte activation and cytotoxicity.

The targeting of tumor cells by cytotoxic T lymphocytes is a promising strategy for biotherapy, but T cells require 2 signals via the T-cell receptor - CD3 complex and CD28 molecules for activation. To bridge the gap between cytotoxic T lymphocytes and tumor cells, our objective in this study was to describe the construction and the cell surface-anchored expression of a fusion protein, anti-CD3 scFv-B7.1, derived from inserting a fusion gene encoding anti-CD3 scFv and the extra-cellular domain of B7.

[In situ analysis of distribution of immunocompetent cells in tumor's local draining lymph nodes].

Aim: To observe the distribution of immunocompetent cells (ICCs) in tumor's local draining lymph nodes (LDLN) at different stages of disease including non-metastasis, micro-metastasis and late metastasis.

Methods: 71 LDLN from 22 breast cancer, 28 LDLN from 7 gastric carcinoma were analysed by using catalyzed signal amplification (CSA) immunohistochemical staining. Monoclonal antibodies (mAbs) to perforin, granzyme B, CD8, CD56, CD68, S-100, CD134 and CD25 were used to detect the amount and functional change of ICCs.

[Immunopotentiation of novel adjuvant SWZY on the poorly immunogenic murine melanoma vaccine].

Aim: To evaluate the immunopotentiation of novel adjuvant SWZY on the cancer vaccine of poorly immunogenic melanoma.

Methods: C57BL/6 mice immunized with inactivated D5 melanoma cells were divided into 6 groups: the control without adjuvants, with FCA, FCA+IL-2+GM-CSF, FIA+IL-2+GM-CSF, FIA+SWZY, and FIA+SWZY+IL-2+GM-CSF. Three days after completion of immunization, DTH response, specific killing activity of splenocytes and the level of IFN-gamma and IL-10 in serum and splenocytes' culture supernatant were assayed in half of the mice in each group.

[Expression of IgVH and B7-1 in proteome of the human colorectal carcinoma cell lines].

Objective: To conduct a proteomic analysis of human colorectal carcinoma cell lines LS174T and SW480 by two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).

Methods: The total proteins of human colorectal carcinoma cell lines LS174T and SW480 were separated with 2-DE using immobilized pH gradient strips and analyzed by MALDI-TOF-MS to obtain peptide mass fingerprints (PMFs). Proteins were identified by using Mascot software to search protein databases.

[The in situ analysis of cytokine mRNA expression in immunological microenvironment about non-small cell lung cancer].

Aim: Using Non-small cell lung cancer (NSCLC) as subject, to explore the characteristics of immune response in immunological microenvironment at the tumor site and its effect on anti-tumor immunity.

Methods: Using in situ hybridization (ISH), the expressions of IL-2, INF-gamma, IL-12 (p40), IL-18, IL-4, IL-10, TGF-beta1, IL-1, IL-3, IL-8, GM-CSF, TNF-alpha and TGF-alpha mRNAs in lymphocytes and tumor cells from five fresh pleural effusion samples and 18 tumor tissue samples of NSCLC patients were detected by in situ hybridization with digoxin end-labeled oligonucleotide probe, using 5 tuberculous pleurisy patients as control.

Results: In pleural effusion and tumor tissue of NSCLC patients, the expression levels of IL-4, IL-10, TGF-alpha, and TGF-beta1 mRNAs were significantly higher than those of IL-2, IL-12, IL-18 and INF-gamma mRNAs.

B7 molecule mRNA expression in colorectal carcinoma.

Aim: To observe the status of tumor-associated B(7) molecule mRNA expression in human colorectal cancer tissue by in situ hybridization.

Methods: The mRNA expression patterns of cancer-associated B(7-1),B(7)H(1),B(7)H(2),ICOS in 22 specimens of human colorectal cancer tissue were monitored by in situ hybridization (ISH) with digoxin-labeled oligonucleotide probes.

Results: B(7-1),B(7)H(1),B(7)H(2),ICOS mRNA were detected in both cancer cells and tumor infiltrating lymphocytes (TIL).

Identification of proteins of human colorectal carcinoma cell line SW480 by two-dimensional electrophoresis and MALDI-TOF mass spectrometry.

Aim: To conduct the proteomic analysis of human colorectal carcinoma cell line, SW480 by using two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption /ionization-time of flight mass spectrometry (MALDI-TOFMS).

Methods: The total proteins of human colorectal carcinoma cell line, SW480 were separated with 2-DE by using immobilized pH gradient strips and visualized by staining with silver nitrate. The gel images were acquired by scanner and 2-DE analysis software, Image Master 2D Elite.

[Study of immune responses induced by human papillomavirus type 18 L1-E6 and L1-E7 chimeric gene DNA vaccines in mice].

Aim: To examine the humoral and cellular immunoresponses induced by HPV18 L1-E6 and L1-E7 chimeric gene DNA vaccines in mice.

Methods: 54 BALB/c mice were divided into 9 groups randomly, and then vaccinated with various recombinant plasmids(pVAX1-L1-E6M3 or pVAX1-L1-E7M3) and immune adjuvants (pLXHDmB7-2 or LTB) through different administration routes (intramuscular or intranasal). After the third inoculation, blood samples were taken to measure specific antibody, and footpad swelling test was used to detect delayed-type hypersensitivity(DTH).

[Eukaryotic expression of anti-CD3 single chain Fv antibody gene and the characterization of its bioactivities].

Aim: To express anti-CD3 scFv in Hela cells and investigate its biological activity.

Methods: DNA fragment encoding anti-CD3 scFv was inserted into eukaryotic expression vector pDisplay. The recombinant expression vector was sequenced and then transfected into Hela cells by electroporation method.

[Construction and expression of eukaryotic expression plasmid pcDNA3.1/hIL-18].

Aim: To construct an eukaryotic expression plasmid pcDNA3.1/hIL-18 and express it in mammalian cells.

Methods: cDNA encoding mature hIL-18 was cleavaged by enzyme digestion from mesomeric clone vector pGEM-T/hIL-18 and inserted into an eukaryotic expression plasmid pcDNA3.

[Study on immunoregulatory functions of E.coli heat-labil enterotoxin B subunit].

Aim: To explore the immunoregulatory functions of Escherichia coli heat-labil enterotoxin B subunit(LTB) and its possible mechanism.

Methods: The rLTB in engineering bacteria VSP60 was purified by Sephacryl S-100 gel filtration chromatography. BALB/c mice were immunized nasally with hen egg lysozyme(HEL) alone or in combination with various doses of LTB.

Expression of Human Single Chain Interleukin 12 Gene in Drosophila Cells.

The hscIL-12 gene(no signal sequence), amplified by PCR, was subcloned into an expressing vector pMT/Bip/V5. The recombinant plasmid was transferedinto Drosophila cell line S2. The secreted hscIL-12 fusion protein was identified by Western blot, and the bioactivity of hscIL-12 was investigated by lymphocyte proliferation assay.