PurposeIt has previously been found that valgus hindfoot alignment (HFA) improves 3 weeks following total knee arthroplasty (TKA) for varus knee osteoarthritis (OA). In the present study, HFA was evaluated prior to TKA, as well as 3 weeks and 1 year following TKA. Using these multiple evaluations, the chronological effects of TKA on HFA were investigated.MethodsThe study included 71 patients (73 legs) who underwent TKA for varus knee OA. Radiograph examinations of the entire limb and hindfoot were performed in the standing position prior to TKA, as well as 3 weeks and 1 year following TKA. The varus–valgus angle was used as an indicator of HFA in the coronal plane. Patients were divided into two groups according to the preoperative varus–valgus angle: a hindfoot varus group (varus–valgus angle <76°) and a hindfoot valgus group (varus–valgus angle ≥76°). The changes in the varus–valgus angle were evaluated and compared in both groups.ResultsIn the hindfoot valgus group, the mean ± standard deviation varus–valgus angle significantly declined from 80.5 ± 3.1° prior to TKA to 78.6 ± 3.7° 3 weeks following TKA and 77.1 ± 2.7° 1 year following TKA. However, in the hindfoot varus group, the mean varus–valgus angle prior to TKA (72.7 ± 2.6°) did not differ significantly from the mean varus–valgus angles 3 weeks (72.3 ± 3.3°) or 1 year (73.5 ± 3.0°) following TKA.ConclusionsHFA improved chronologically in legs with hindfoot valgus as a result of the alignment compensation ability of the hindfoot following TKA. However, no improvement was noted in legs with hindfoot varus because the alignment compensation ability of the hindfoot had been lost. The patients with hindfoot varus should be attended for ankle pain in the outpatient clinic after TKA.Level of evidenceIII.

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The present system may provide a useful means of phenotypic analysis of genetic information in mammalian organs for basic research as well as therapeutic molecular targeting in the post-genomic era.

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Objective. To deliver and overexpress the hsp70 gene in cultured chondrocytes to investigate its effect on nitric oxide (NO)-induced apoptosis of chondrocytes.Methods. Primary chondrocyte cultures were established from rabbit joints. The cells were transduced with an empty adenovirus vector (Ax1w) or an adenovirus vector harboring the hsp70 E-tag fusion gene (AxSHEwt Degradation of articular cartilage, the major component of a joint, causes serious dysfunction of joints. Osteoarthritis (OA) is associated with degeneration of articular cartilage, but its pathogenesis has not yet been fully elucidated. Various biologic and chemical stress factors are thought to be involved in the occurrence and progression of OA, including mechanical stress, inflammatory cytokines such as interleukin-1 (IL-1) and tumor necrosis factor ␣, and reactive oxygen species (ROS) such as nitric oxide (NO). Chondrocytes are responsible for the maintenance of articular cartilage. Reduced cartilage cellularity has been postulated to play a role in the pathogenesis of OA (1,2), and the reduction in cellularity in human OA cartilage is partly attributable to apoptosis, i.e., programmed cell death. Recent studies have shown apoptosis of articular chondrocytes in OA in humans and in experimental animals (3,4), which implies the importance of apoptosis in the progression of the pathology of OA. Thus, apoptosis has attracted the attention of researchers as a potential etiologic factor in OA progression (5). If apoptosis of OA chondrocytes could be controlled, the implications of apoptosis in OA pathogenesis would become clearer, and the suppression method would suggest a novel therapeutic intervention against OA.One of the major cellular responses to stress involves synthesis of a set of highly conserved proteins, called heat-shock proteins (HSPs), during a process referred to as the stress response. In mammalian systems, HSPs include proteins with molecular weights of 110, 90, 70, 60, 40, and 27 kd. Some HSP members are constitutively expressed, and the expression levels increase in response to stress, while others are induced after exposure to stress. HSPs not only protect cells from stress-induced damage, but also facilitate the recovery of cells from the damage by binding to degraded or misfolded polypeptides (6-8). The inducible Hsp70 is a major HSP, and its expression is augmented in OA

Naked plasmid DNA (pDNA) injection could become an alternative procedure to viral and nonviral gene delivery systems. We have previously shown that Epstein-Barr virus (EBV)-based plasmid vectors containing the EBV nuclear antigen 1 (EBNA1) gene and the oriP sequence enable quite high and long-lasting expression in various in [9][10][11] and to a lesser extent, in the spleen, kidney, lung and heart. 9 The expression levels critically depended on the injection volume and rate, strongly suggesting that the 'hydrodynamic' pressure plays a key role in the genetic transfer. The Epstein-Barr virus (EBV)-based plasmid vector carries two genetic elements from EBV, ie the EBNA1 gene and the oriP element. 12 The EBNA1 binds to oriP in a sequence-specific manner and exerts various effects on the oriP-bearing plasmid. In human cells, the plasmid replicates in synchrony with chromosomal DNA, resulting in long-term retention and expression. 12 The EBNA1 gene and oriP have been employed to engineer artificial chromosomes. 13,14 On the other hand, the EBNA1 also facilitates nuclear localization of the plasmid, 15,16 binding of plasmid to the nuclear matrix, 17 and transcriptional up-regulation. [18][19][20] Collectively, the EBVbased plasmid vector gives not only a prolonged, but also a stronger expression of the transgene. We have applied the EBV-plasmids to various preclinical studies of gene therapy, including those for hereditary, 21 chronic, 22,23 as well as malignant diseases. [24][25][26][27] In some of these studies, the EBV-based plasmid vectors were combined with cationic liposomes, 25,28 cationic polymers, 23,[25][26][27]29 electroporation 21 or gene gun, 30 while in the other studies, naked EBV-based plasmid was injected into the cardiac muscle. 22,29 In the present study, we transferred the naked EBV-based plasmid vectors to the liver through the intravascular route.EBV-based and conventional plasmid vectors encoding the luciferase or ␤-gal genes as markers (Figure 1, upper panels) were constructed and injected intravenously into the tail vein of mice as described. 9 The injection with pG.GL3, a conventional luciferase-expression vector,

The EBV/lipoplex is a nonviral gene delivery system composed of a cationic lipid and Epstein-Barr virus (EBV)-based plasmid vector that carries the EBV oriP and EBV nuclear antigen 1 (EBNA1) gene. Because the EBNA1 supports retention, nuclear localization, and transcriptional upregulation of the oriP-bearing plasmid, cells transfected with the EBV/lipoplex express the transgene at a very high level. We hypothesized that tumor cells genetically manipulated with the EBV/lipoplex may be used as a tumor vaccine without drug selection, strongly contributing to immunotherapy of patients with malignancies. The cytokines interleukin (IL)-12 and IL-18 exert a variety of immune-regulatory functions including interferon (IFN)-gamma production and cytotoxic T lymphocyte (CTL) and natural killer (NK) activation. Here, we investigated the possible therapeutic effects of an autologous tumor cell vaccine in the B16 melanoma model. The vaccine was engineered to secrete IL-12 and IL-18 by means of the EBV/lipoplex. B16 cells were subcutaneously implanted into syngenic mice followed by repetitive immunization with irradiated B16 cells that had been transfected 3 days earlier by TFL2-3, a novel cationic lipid, with EBV-plasmid vectors encoding IL-12 and/or IL-18 genes (B16/mIL-12, B16/mIL-18, and B16/mIL-12+mIL-18). The mice vaccinated with B16/mIL-12 underwent strong tumor suppression accompanied by a high IFN-gamma production. Both CTL and NK activities were significantly elevated in these mice. When the tumor cell vaccine was prepared by means of conventional (non-EBV) plasmid vectors combined with the same cationic lipid, the therapeutic outcome was not as good, suggesting the superiority of the EBV-based plasmid in engineering these types of tumor vaccines. Vaccination with B16/mIL-18 was not effective in suppressing tumors, whereas B16/mIL-12+mIL-18 showed comparable antitumor therapeutic validity as B16/mIL-12 did. When IFN-gamma mutant (IFN-gamma(-/-) mice were treated, B16/mIL-12 vaccine did not show any therapeutic activity, suggesting the necessity of IFN-gamma in the anti-melanoma immune responses. In contrast, the antitumor effect was not affected by NK depletion in mice that received repetitive injections with anti-asialo GM1 antibody. Furthermore, vaccination with B16/mIL-12 significantly suppressed pulmonary metastases in mice that had been intravenously injected with parental B16. Our results suggest that the EBV/lipoplex is quite useful in generating an autologous tumor cell vaccine and that IL-12 is an important component of the vaccine.