Targeted hepatocellular carcinoma proapoptotic BikDD gene therapy.

Hepatocellular carcinoma (HCC), the third leading cause of cancer death in the world, is the most general type of primary liver cancer. Although current treatment modalities, such as liver transplantation, resection, percutaneous ablation, transarterial embolization, chemotherapy and radiotherapy are potentially curative, these methods are not universally applicable to all of HCC patients, especially for those with poor prognosis in which no effective remedy is available. Therefore, development of novel therapeutic approach for the treatment of HCC is urgently needed. In the current study, we developed a promising HCC-targeted gene therapy vector driven by liver cancer-specific α-fetoprotein promoter/enhancer coupled to an established platform technology. The activity of this expression vector is comparable with or even higher than that of strong cytomegalovirus (CMV) promoter and exhibits strong promoter activity in liver cancer cells/tumors, but has nearly no or very low activity in normal cells/organs in vitro and in orthotopic animal models in vivo. Its cancer specificity exceeds that of the CMV promoter, which expresses non-specifically in both normal and tumor cells. In addition, targeted expression of a therapeutic BikDD, a mutant of proapoptotic gene Bik effectively and preferentially killed liver cancer cells, but not normal cells and significantly repressed growth of HCC tumors, and prolonged survival in multiple xenograft and syngeneic orthotopic mouse models of HCC through intravenous systemic gene delivery. Importantly, systemic administration of BikDD by our expression vector exerted no systemically acute toxicity compared with CMV-BikDD in mice. Taken together, this study elucidates a relatively safe and highly effective and specific systemic gene therapy strategy for liver cancer, and is worthy of further development for future clinical trials.

Deficient transforming growth factor beta and interleukin-10 responses contribute to the septic death of burned patients.

In order to understand the roles of pro-inflammatory and anti-inflammatory cytokines in burn injury and sepsis post-burn, serial changes in serum levels of transforming growth factor beta-1 (TGF-beta-1) were determined and compared to those of IL-6 and IL-10 in 15 burned patients. Among these 15 patients, 8 recovered without sepsis. The other seven, who were septic, expired. Our results showed that an initial peak serum TGF-beta-1 response was detected within 1 day post-burn. Peak serum IL-6 and IL-10 responses were also detected within 4 days after the burn injury of these patients. Significant differences in peak serum IL-6, IL-10 and TGF-beta-1 levels were not found between patients with total body surface area (TBSA) of greater or less than 50% and between patients who survived or expired from burn injury. Afterwards, levels of circulating IL-6 and IL-10 remained low in the survivors. However, a second peak response in serum TGF-beta-1 levels was observed in all burned patients analyzed. The second peak serum TGF-beta-1 levels post-burn of the eight survivors and the seven non-survivors were from 28,542 to 76,554 pg/ml (a mean value of 51,256+/-14,264 pg/ml) and from 8616 to 40,851 pg/ml (a mean value of 24,079+/-10,399 pg/ml), respectively. A significant difference (P<0.01) in mean values of the second peak TGF-beta-1 responses between groups of survivors and non-survivors was detected. Levels of circulating IL-6 in the septic non-surviving patients showed a tendency to increase 1-2 weeks post-burn and reached high levels before the expiration of these patients. After an initial peak response, the serum IL-10 level remained low in one of the seven non-survivors, while it increased in the other six non-survivors. However, marked increases in circulating IL-10 levels were observed only just before the death of these non-survivors. In conclusion, an initial increase in serum levels of IL-6, IL-10 and TGF-beta-1 was detected post-burn. A marked increase in serum levels of IL-6 before death suggests its role in the pathophysiology of sepsis in burned patients. In addition, a low secondary TGF-beta-1 response and a lack and/or delay in the increase of circulating IL-10 in the non-survivors may all contribute to the pathophysiology of septic death in burned patients.

Necrotizing fasciitis of the head and neck: an analysis of 47 cases.

Necrotizing fasciitis is an overwhelming infection common to the perineum, abdominal wall, and extremities. It is a surgical emergency related to a high mortality rate that is more often seen in elderly and immunocompromised patients. Necrotizing fasciitis occurs uncommonly in the head and neck region. Over a 12-year period, 47 cases of necrotizing fasciitis of the head and neck region were collected at this hospital. The demographics, predisposing factors, clinical presentation and courses, management, complications, and outcomes were analyzed. The cases were divided into two groups: survivors and nonsurvivors. Statistical comparisons were made of the parameters age, gender, smoking or drinking habit, underlying medical problems, laboratory data, and treatments used. Forty-two patients (89.4 percent) had associated systemic disease; most of these patients had diabetes (72.3 percent). The clinical manifestations are nonspecific but are often typical for diagnosis. The necessity of computed tomographic scans is not conclusive in this study. Presentation of septic shock (p = 0.004) and association with underlying malignancy (p = 0.03) were the only statistically significant factors that led to a poor prognosis. The cornerstones of proper management include early diagnosis, aggressive surgical debridement, broad-spectrum antibiotics, and intensive supportive care.

Detection of a spontaneous high expression of heat shock protein 70 in developing zebrafish (Danio rerio).

A spontaneous high expression of heat shock protein 70 (HSP 70) was detected in zebrafish (Danio rerio) at early larval stage (84 h after fertilization), but the HSP 70 level was either low or barely detectable in 12, 24, 36, 60, and 108 h after fertilization. The extracts of zebrafish at 80 and 84 h after fertilization formed a clear protein-DNA complex with a probe containing heat shock elements (HSEs), suggesting that this spontaneous expression of HSP 70 may be turned on via the binding of stage-specific HSE-binding factors to HSP 70 gene promotor. The protein-HSE complexes produced by the spontaneous binding, however, were found to be different from those formed by the extracts of heat-treated zebrafish in electrophoretic mobility.

Changes in circulating levels of an anti-inflammatory cytokine interleukin 10 in burned patients.

In order to understand the role of an anti-inflammatory cytokine interleukin 10 (IL-10) in the pathophysiology of burn injury, IL-10 levels in serial serum samples of 22 burned patients were analyzed. The total body surface areas (TBSA) of the burn injury ranged from 30 to 90%. Among these 22 patients, 14 recovered and the other eight, who were septic, expired. A significant difference in serum IL-10 values on admission (5-20 h postburn) was found (P<0.05) between patients who survived or died from burn injury as analyzed by the Student's t test. In addition, a significant difference in serum IL-10 on admission was also found (P<0.05) between patients with TBSA of greater or less than 50%. An initial peak serum IL-10 response was detected within 2.5 days postburn. Significant differences in the peak serum IL-10 levels were not found between patients with TBSA of greater or less than 50% and patients who survived or expired from burn injury. Afterwards, serum IL-10 remained low in the survivors, while an increase in serum IL-10 could be detected in the non-survivors with proven sepsis. Levels of circulating IL-6 in these non-surviving patients showed a tendency to increase starting from about 1-2 weeks postburn which coincided temporally with the detection of infections. However, marked increases in circulating IL-10 levels were observed just before death in four of the eight non-survivors. The serum samples of these four patients were collected at 31 h (404.8 pg/ml), 2 h (773.9 pg/ml), 5 days (150.7 pg/ml) and 12 h (177.1 pg/ml) before the expiration of these patients, respectively. IL-10 levels of 28.6, 27. 5 and 13.5 pg/ml were detected in sera of three of the remaining four non-survivors that were collected at 2.5 h, 36 h and 30 h before the expiration of these patients, respectively. There was one non-surviving patient who suffered an 80% burn (patient D4 in Table 1 and Fig. 4) and his IL-10 level at 20 days postburn was 13.4 pg/ml. The serum sample of this patient was collected 22 days before death and he was not suffering from sepsis at this stage. In conclusion, an initial increase in serum levels of IL-10 was detected postburn. A marked increase in serum levels of IL-10 was detected in four of the eight septic patients just before their death. It was considered that a lack and/or a delay in the increase of circulating IL-10 may play a significant role in the pathophysiology of sepsis in burned patients.

Changes in circulating levels of interleukin 6 in burned patients.

Interleukin 6 (IL-6) levels in serial serum samples of 10 burned patients were analyzed. The total body surface areas (TBSA) of the burn injury varied from 30 to 85%. Among these 10 patients, five recovered and the other five, who were septic, expired. A significant difference in serum IL-6 values on admission (5-13 h postburn) was found (p < 0.05) between patients who survived or died from burn injury as analyzed by the Wilcoxon's rank sum test. In addition, a significant difference in serum IL-6 on admission was also found (p < 0.05) between patients with TBSA of greater or less than 50%. Afterwards, an initial peak serum IL-6 response was detected within 4 days postburn. Significant differences in the peak serum IL-6 levels were not found between patients with TBSA of greater or less than 50% and patients who survived or expired from burn injury. In the survivors, serum IL-6 remained low, while IL-6 increased markedly starting at about one to two weeks postburn in four of the five nonsurvivors with proven sepsis. Except for the patient who expired 42 days postburn, the maximum serum IL-6 values of the other four nonsurvivors were all greater than those of the five survivors from burn injury. Significant correlation (p < 0.05) relating the change in serum IL-6 and body temperature was observed in only two (one survivor and one nonsurvivor) of the ten patients. Changes in serum IL-6 were also compared with changes in circulating TNF-alpha and IL-8 determined previously. A similar pattern in the dynamic changes of circulating TNF-alpha, IL-8 and IL-6 was observed in the individual burned patient. An increase in serum levels of all three cytokines was detected postburn. Serum levels of three cytokines were significantly higher in the septic patients, who all died. It was considered that all three cytokines analyzed may play a significant role in the pathophysiology of sepsis in burned patients.

Changes in serum tumour necrosis factor-alpha in burned patients.

Dynamic tumour necrosis factor-alpha (TNF-alpha) changes in serial serum samples of 10 burned patients were analysed in this study. The total body surface areas (TBSA) of the burn injury were from 30 to 85 per cent. Among these 10 patients, five recovered and another five died with proved sepsis. On admission which was about 5-13 h postburn, eight of the 10 patients showed their serum TNF-alpha levels to be higher than the mean serum TNF-alpha value of five healthy laboratory personnel. Furthermore, an initial peak serum TNF-alpha response which could be detected within 2.5 days after burn injury has also been observed. However, significant differences in both the serum TNF-alpha values on admission, as well as the first peak serum TNF-alpha levels, were not found (P > 0.05) between patients with TBSA of greater or less than 50 per cent and patients who survived or died from burn injury. In the survivors, serum TNF-alpha stayed at low levels, while it increased markedly in four of the five non-survivors with proven sepsis starting at about 1 week postburn. A significant difference in the maximum serum TNF-alpha levels (P < 0.05) was detected between patients who recovered and died from the thermal injury. In conclusion, great increases in serum TNF-alpha levels have been detected in burned patients with the occurrence of bacterial infection postburn. It is suggested that strategies for the inhibition of TNF-alpha production or in the neutralization of TNF-alpha activity should also be considered in the better treatment of burned patients.

Changes in levels of serum IL-8 in burned patients.

Interleukin (i.l.)-8 levels in serial serum samples of 10 burned patients were analysed. The total body surface areas (TBSAs) of the burn injury ranged 30 to 85 per cent. Of these ten patients, five recovered and the other five, who were septic, died. On admission at about 5-13 h postburn, one of the five survivors and two of the non-survivors had serum IL-8 levels higher than 18.1 pg/ml, which is the detection limit of the IL-8 assay kit. The serum IL-8 values of six healthy laboratory personnel included in the present study were all less than 18.1 pg/ml. Afterwards, an initial peak serum IL-8 response was detected within 2-4.5 days postburn. Significant differences in the peak serum IL-8 levels were not found between patients with TBSAs of greater or less than 50 per cent and patients who survived or expired from burn injury. In the survivors, serum IL-8 remained low, whereas IL-8 increased markedly, starting at about one week postburn in four of the five non-survivors with confirmed sepsis. Significant differences in the maximum serum IL-8 levels were detected between patients who recovered vs. those who died from the thermal injury. In conclusion, the results showed that there was an increase in serum IL-8 postburn. Serum IL-8 was significantly higher in the septic patients, who all died. This cytokine may play a significant role in the pathophysiology of sepsis in burned patients.

Coverage of chest wall defect with pectoralis-breast myocutaneous flap: a report of two cases.

Reconstruction of a chest wall defect remains a challenging problem, especially where such defects are radiation related. Two cases of modified breast flaps associated with pectoralis major muscle were performed to cover a huge mediastinal defect, with heart lung exposure and a post-mastectomy radiation-related chest wall defect with partial rib resection. Both modified breast flaps healed uneventfully. No respiratory distress was encountered post-operatively. The breast flap associated with pectoralis major muscle ensured flap reliability and minimized flap complication. It is easy to raise and its size is large enough to cover of large chest wall defect which is radiation related, or those internal mammary arteries and thoracodorsal arteries compromised in previous surgery where the transverse rectus abdominis myocutaneous (TRAM) flap and latissimus dorsi flap become unreliable. In such cases a contralateral modified breast flap associated with pectoralis major muscle is the better choice.

Surgical treatment of axillary osmidrosis: an analysis of 343 cases.

From July of 1986 to July of 1992, 343 patients have received surgery for axillary osmidrosis by partially removing skin and cellular tissue en bloc and removing the subcutaneous cellular tissue of the adjacent area. A total of 102 patients were followed for 4 months to 6 years, with an average of 32 months. The total satisfaction rate was 91 percent (93 of 102). The wound complication rate was 6.715 percent (46 of 685). There were no scar contractures or limitations of arm abduction. In this paper we emphasize three merits of our procedure. One is that partially removing the skin promises definite excision of more than half the eccrine glands which were located in the dermis of the operative field. The second merit is good exploration for undermining and defatting of the under-surface of the adjacent area. The third merit is a low wound complication rate because the width of the skin excision is less than 3 cm. Therefore, partial removal of skin and cellular tissue en bloc and the subcutaneous cellular tissue of the adjacent area is the choice for surgical treatment for axillary osmidrosis.

Bacteriology of necrotizing fasciitis: a review of 58 cases.

Necrotizing fasciitis is a relatively rare soft tissue infection that is characterized by rapidly widespread necrosis of the fascia and subcutaneous tissue. Fifty-eight cases were reviewed over a 5.5-year period. The majority of wound cultures were polymicrobial and single culture rate was 18.6%. The facultative organisms were most frequently recovered. The most frequently discovered bacteria were E. coli, Proteus mirabilis, Klebsiella pneumoniae, Staphylococcus aureus and group D enterococcus. The overall mortality rate was 48.3%. Early recognition, serial radical surgical debridement and strong and appropriate antibiotics were essential for survival.

Comparisons of different animal skins with human skin in drug percutaneous penetration studies.

The in vitro penetration of theophylline, sodium diclofenac and benzoic acid through artificial cellulose membrane and eight animal skins was investigated. The intact animal skins including stratum corneum (SC) and viable epidermis were taken from frog, snake with or without scales, nude mice, Sprague-Dawley rat, porcine and human prepuce and thigh skin. The results indicated that the penetration was fastest through cellulose membrane and frog skin and slowest through human prepuce and thigh skin. The snake skin with scales slowed down the penetration rate more significantly than the scaled skin. Benzoic acid was the fastest penetrant through all animal skins. The permeable behaviors of sodium diclofenac through SC and intact skin of snake, porcine and human were compared. In porcine, sodium diclofenac penetrated through SC at a rate 33 times higher than through intact skin, but in snake and human skin, the rate through SC was only 2.2 and 1.6 times higher than through intact ones. This implies that both viable epidermis and SC were the major rate limiting barriers in drug penetration. DSC thermograms and IR spectra showed that the SC of snake, porcine and human thigh were very similar in structure and components. The study suggests that snake skin, porcine skin and human prepuce skin could replace the human skin in in vitro drug penetration experiments.

[Clinical application of Chinese forearm flap: 5-year review].

Twenty-nine patients treated with the Chinese forearm flap from 1985 to 1989 were reviewed. This flap was used for soft tissue defect reconstruction in 19 cases, and phallus reconstruction in 10 patients. There were two cases of total flap failure and two cases of partial failure. Twenty cases used sensory flaps. Donor site complications included partial loss of the skin graft with tendon exposure in one patient (3.4%), limitation of range of movement of the wrist joint in one case (3.4%), and paresthesia over the radial site of the hand in 12 patients. On functional evaluation, 20 patients demonstrated no significant loss of strength in the donor extremity.

Comparison of scar contracture with the use of microskin and Chinese-type intermingled skin grafts on rats.

Wounds that were measured precisely 4 x 5 cm in size were created over the dorsal surfaces of rats. The defects were grafted with the use of either the microskin or Chinese technique of intermingled auto/allograft with an expansion ratio of 10:1. The size of the grafted wound of each rat was recorded on the fifth graft day after grafting and then weekly for 11 weeks. The extent of scar contracture on each rat was calculated weekly as a percentage of the original size. The results showed that the scar contracture associated with the Chinese type of intermingled auto/allograft transplantation was less than that associated with the microskin technique. The healing process in both forms of grafting was uneventful.