Human Systems Therapy:Use of Interpersonal Relations in Promoting Therapeutic Change

The aim of this work is to present the central role of interpersonal relations in Human Systems Therapy(HST).Psychotherapies can be classified as(a)down-top therapies,such as individual therapies following reductionism,namely influencing the whole(behavior)through the intra-psychic parts,(b)top-down or holistic therapies,such as family therapies influencing the parts(individuals)through the whole family,and(c)the combination of down-top and top-down.Emotionally Focused Therapy(EFT)and Multisystemic Therapy(MST)act on two levels that influence each other,namely the individuals and their relations.This is also applied,more or less,on relational and interpersonal therapies.When there is an intervention in two interacting parts or levels,then the result of the intervention is more than additive and more effective.When the levels are more than two,then the effectiveness is expected to be even higher.This is the case of Human Systems Therapy(HST),which is applied intrapsychically,on interpersonal relations and on family and/or larger systems,with interpersonal relations having a central role.HST is based on general systems principles and it constitutes a multilevel approach:(a)a definition of a human system,(b)epistemology,(c)a model of human systems personality/intelligence,(d)an increase in order and variety.Besides,HST includes a large number of techniques.Empirical support of HST effectiveness shows that(a)after seven sessions with schizophrenics,39%of psychotic symptoms are reduced,(b)after eight sessions with hashish adolescent users,all users stop the use completely,and(c)9 out of 11 psychiatric non-psychotic cases(81%),after four sessions,become non-psychiatric ones(treated).

Disease Prevention and Alleviation by Human Myoblast Transplantation

Myoblast implantation is a unique, patented technology of muscle regeneration being tested in Phase III clinical trials of muscular dystrophy, ischemic cardiomyopathy, Phase II trial of cancer, and Phase I trial of Type II diabetes. Differentiated and committed, myoblasts are not stem cells. Implanted myoblasts fuse spontaneously among themselves, replenishing genetically normal myofibers. They also fuse with genetically abnormal myofibers of muscular dystrophy, cardiomyopathy, or Type II diabetes, transferring their nuclei containing the normal human genome to provide stable, long-term expression of the missing gene products. They develop to become cardiomyocytes in the infracted myocardium. Myoblasts transduced with VEGF165 allow concomitant regeneration of blood capillaries and myofibers. They are potent biologics for treating heart failure, ischemic cardiomyopathy, diabetic ischemia, erectile dysfunction, and baldness. Myoblasts, because of their small size, spindle shape, and resilience, can grow within wrinkles and on skin surfaces, thus enhancing the color, luster and texture of the skin “plated” with them. They can be injected subcutaneously as a cellular filler to reduce wrinkles. Intramuscular injection of myoblasts can augment the size, shape, consistency, tone and strength of muscle groups, improving the lines, contours and vitality to sculpt a youthful appearance. This highly promising technology has great social economic values in treating hereditary, fatal and debilitating disease conditions.

ESTABLISHMENT OF K562/ADM/VER CELL SUBLINE RESISTANT TO VERAPAMIL AND ITS RESISTANT MECHANISM

Objective: To understand whether verapamil (VER) resistance development in the multidrug-resistant cell line and its mechanism. Methods: K562/ADM/VER cell subline resistant to verapamil was established through a gradual increase of VER concentration in the media. MTT method was used to assay resistance to VER, cross resistance to dipyriamole (DPM), cyclosporin A (CsA) in the cells, and HPLC and spectrofluorometer to detect intracellular accumulation of VER or ADM respectively, as well as S-P immunocytochemical technique for detection of genes expression. Results: It were observed that 7.9—fold increase in VER resistance, significantly reduced intracellular accumulation of VER or ADM and also develop across resistance to DPM and CsA in K562/ADM/VER cells, compared with its parent cell, K562/ADM. High-level of p-glycoprotein(pgp), middle-level of p53, p16, was present in two cell lines without expression of GSTPI, C-myc, C-myc, C-fos and C-erbB-2. Bc1–2 protein expression was found only in K562/ADM cells. Conclusion: K562/ADM cells were capable of being induced to develop resistance to VER.

Tumor Antigen Specific Activation of Primary Human T-Cells Expressing a Virally Encoded Chimeric T-Cell Receptor Specific for p185HER2

We have developed and tested chimeric T-cell receptors (TCR) specific for p185HER2. In these experiments, retroviral vectors expressing the N29γ or N29ζ receptors were constructed in pRET6. Amphotropic viral producer cells were established in the GALV-based PG13 packaging cell line. Ficoll purified human peripheral blood lymphocytes (PBL) were virally transduced using an optimized protocol incorporating activation with immobilized anti-CD3/anti-CD28 monoclonal anti- bodies, followed by viral infection in the presence of fibronectin fragment CH296. Transduced cells were co-cultured with human tumor cell lines that overexpress (SK-OV-3) or underexpress (MCF7) p185HER2 to assay for antigen specific im- mune responses. Both CM+ and CD8+ T-cells transduced with the N29γ or N29ζ chTCR demonstrated HER2-specific anti- gen responses, as determined by release of Th1 like cytokines, and cellular cytotoxicity assays. Our results support the fea- sibility of adoptive immunotherapy with genetically modified T-cells expressing a chTCR specific for p185HER2.

Mature insulin production by engineered non-βcells

To pursue insulin and islet transplantation replacement therapy for type 1 diab etes based on engineered human non β cells which secrete mature insulin Methods Human proinsulin cDNA was cloned from its genomic gene and mutated by overlap e xtension PCR, introducing furin consensus cleavage sequences (Arg Xaa Lys/Arg Arg) An expression vector encoding a genetically modified human proinsulin c DNA was generated and transduced to Hela, 293, and L02 cells by lipofectin medi ated DNA transfection Following G418 screening, the surviving L02 cells were s elected and enriched Insulin levels in the supernatant and cells were evaluate d using radioimmunoassay and immunofluorescence staining Results Three sites in the insulin gene were mutated simultaneously Insulin gene m odified cells were able to express insulin at different levels: 8 45-188 00? μIU/24 h/2 0×10 6 Hela cells and 159 88-242 14?μIU/24 h/2 0×10 6 293 cells for transient expression, and 2 56-61 95?μIU/24 h/2 0×10 6 from se veral L02 clones screened with G418 No insulin was released by control cells Furthermore, immunofluorescence staining confirmed that proinsulin was stored a s vacuoles in the cytoplasm of L02 cells Conclusion A correctly mutated human proinsulin cDNA was obtained successfully, transfected and expressed efficiently in non beta cells, lending support to the study of s omatic gene therapy in diabetes mellitus

Effects of anti-HPV16E6-ribozyme on phenotype and gene expression of a cervical cancer cell line

To investigate the effects of anti HPV16E6 ribozyme (HRz) on phenotype and gene expression of a cervical cancer cell line Methods HRz was designed by computer programs HRz’s activity was identified by cleavage experiments in vitro HRz and empty eukaryotic plasmids were transfected into CaSKi cells with lipofectin, then renamed CaSKi R and CaSKi P, respectively The expression of ribozyme in transfected cells was observed by RNA dot blot The amounts of E6 mRNA in three kinds of cells lines were detected by Northern blot Cell growth curves and soft agar forming ability were studied The ability of each cell line to form tumors was assessed in nude mice Apoptosis rates and expression of c myc, bcl 2, p53 and Fas were detected by flow cytometry (FCM) Antigens of tumor cells, HLA 1, HLA 2, B7 1 and B7 2 were also detected NK, LAK, and CD 3AK cells were induced Their cytotoxicities were detected in CaSKi R, CaSKi P, and CaSKi cells Results In vitro cleavage reaction demonstrated that HRz could cleave HPV16E6 mRNA in a site specific manner HRz could be expressed stably in transfected CaSKi cells Northern blot analysis showed that E6 mRNA levels were lower in CaSKi R than in CaSKi The growth rate of CaSKi R was slower than those of CaSKi and CaSKi P The soft agar forming rate of CaSKi R was lower compared with those of CaSKi and CaSKi P cells The ability of CaSKi R to form tumors in nude mice was also poor The apoptosis rate of CaSKi R cells was much higher than those of CaSKi and CaSKi P HRz could reduce the expression of E6, c myc and bcl 2 proteins, and increase the expression of p53 as well HRz could increase the expression of HLA 2, B7 1 and B7 2 antigens The cytotoxicity of NK, LAK and CD 3AK cells was much higher in CaSKi R than in CaSKi P and CaSKi cells Conclusion HRz not only reverses the malignant phenotype of CaSKi cells partially, but also induces apoptosis in the cells, and increases sensitivity of CaSKi cells to immune cells