Novel gene therapy for rheumatoid arthritis with single local injection: adeno-associated virus-mediated delivery of A_(20)/TNFAIP_(3)

Dear Editor,Multiple experiments have established TNF-α-induced protein 3 (A20/TNFAIP3) as a critical regulator associated with rheumatoid arthritis(RA)[1,2].The lack of TNF-α-induced protein 3 (A20) promotes the NOD-like receptor protein 3 (NLRP3) inflammasome and induces spontaneous arthritis,while increase of A20 reduces the secretion of IL-1β and favors immunological tolerance[3,4]. Hence, we investigate the feasibility of recombinant adeno-associated virus 6 (rAAV6)-mediated A20 gene therapy in a collageninduced arthritis (CIA) model.

GENE TRANSFER INTO PORCINE MYOCARDIUM VIA PERICARDIAL CAVITY BY HOMEMADE EASY PUNCTURE DEVICE

Objective To explore the feasibility and safety of gene transfer into porcine myocardium via the pericardial cavity by a homemade easy device. Methods Replication-deficient recombinant adenoviral vector carrying LacZ report gene (Ad-LacZ) was constructed by the calcium phosphate precipitation method. Twelve healthy Chinese mini-swine were randomly divided into experimental group (n=6) and control group (n=6). Acute myocardial infarction (AMI) model was established by balloon occlusion of the distal part of D1 branch of left anterior descending (LAD) artery, at the same time the intra-pericardial cavity injections were performed through the small incision of the abdominal wall below the xyphoid appendix using a homemade device. Then gene transfer was performed using a central venous catheter. The pericardium was pretreated with injection of a mixture of collagenase (1 200 U) and hyaluronidase (3 000 U) in both groups. Then 2.0×109 plaque formation unit (PFU) Ad-LacZ was injected into the pericardial cavity in experimental group, while 1 mL of normal saline was injected in the control group. The β-galactosidase activity detection and X-gal staining of the ischemic myocardium were performed on the 3rd, 7th, and 28th day after injection. Results The LAD artery was occluded completely and infarction and ischemia were detected by histological assessment. In experimental group, the X-gal staining positive cells and β-galactosidase activity quantification were detectable on the 3rd day after injection, increased markedly on the 7th day, and then declined on the 28th day. The transfer efficiencies indicated by the positive myocardial cells were 16.7%, 45.6%, 22.8% on the 3rd, 7th, 28th day, respectively. In control group, no positive cells and β-galactosidase activity were observed. Conclusion Adenovirus can be transferred into ischemic myocardium and express target gene in the AMI model for four weeks with the homemade easy device via pericardial cavity pretreated by collagenase and hyaluronidase.

Immunohistochemical Analysis of the Acid Secretion Potency in Gastric Parietal Cells

Gastric parietal cells are important in acid secretion, but it is unclear which cells throughout the gastric gland have the highest secretion potency. Here, we used immunohistochemical methods with anti-H+, K+-ATPase, phosphoryl ezrin and CD44 antibodies to study the distribution of gastric acid secretion activity. Stomach tissues from freely fed and starved rats were cryofixed for light microscopy or fixed by high-pressure freezing for electron microscopy. Parietal cells from freely fed animals corresponded to the active secretion phase and to the inactive resting phase from starved rats. Anti-H+, K+-ATPase and anti-phosphoryl ezrin labeling were observed on the membrane of the intracellular canaliculi and the tubulovesicle from freely fed rats, while cells from starved animals showed weak labeling with anti-phosphoryl ezrin antibody staining. Morphometrical analysis at the electron microscopic level was performed on active and inactive acid secretory phases between the upper and base regions of the gland. H+, K+-ATPase and CD44 were distributed on both sites of the microvillous and tubulovesicle membrane in the same cells, but phosphoryl ezrin localized predominantly on the microvillous membrane in active cells of the glandular neck and upper base. Therefore, the highest secreting potency appeared to be in cells of the glandular neck and upper base.

慢性阻塞性肺病中的树突状细胞旧患的新突破口

树突状细胞（dendritic cells，DCs）是专门的抗原提呈细胞（antigen—presenting cells，APCs），它与免疫稳态（immune homeostasis）有关。在组织损伤或感染的肺部应答中，DCs能启动和统筹先天性免疫和获得性免疫（adaptive immunity）。