[Effects of artificial dermis combined with autologous skin in repairing the wounds with exposed bone and/or tendon in fingers of children after electric burns].

Objective: To explore the effects of artificial dermis combined with autologous skin in repairing the wounds with exposed bone and/or tendon in fingers of children after electric burns. Methods: A retrospective observational study was conducted. From January 2017 to December 2022, 14 children with bone and/or tendon exposed wounds in fingers after electric burns who met the inclusion criteria were admitted to Zhengzhou First People's Hospital, including 9 males and 5 females, aged 2 to 11 years. A total of 38 fingers were affected, with 1 wound per finger. After debridement, artificial dermal coverage combined with vacuum sealing drainage was performed in all the wounds in the first stage, with wound area of 2.0 cm×1.0 cm-4.5 cm×2.5 cm after debridement. The second stage surgery was performed to close the wound with autologous thin intermediate thickness skin graft. Then the children were told to perform functional rehabilitation exercise as early as possible. The survival of autologous skin graft was observed at the 7th day after the second stage surgery. The wound healing time was recorded. After 12 months of follow-up, the Vancouver scar scale was used to evaluate the scar hyperplasia at the skin grafting site of the affected finger; the total action mobility (TAM) of the affected finger joint was measured for evaluating the functional recovery of the affected finger; a self-made efficacy satisfaction rating table was used to investigate the parents' satisfaction with the curative effect of the children. Results: At the 7th day after the second stage surgery, all the children had good survival of autologous skin grafts. The wound healing time was (24.1±2.7) d. After 12 months of follow-up, the scar score at the skin grafting site of the affected finger was 5.2±2.4; the TAM of the affected finger joint was (177±40)°; the functional assessment was good in 12 fingers, medium in 23 fingers, and poor in 3 fingers; the parents' satisfaction with the curative effect of the children in the survey was very satisfied in 10 cases, satisfied in 3 cases, and dissatisfied in 1 case. Conclusions: The combination of artificial dermis and autologous thin intermediate thickness skin graft is an alternative surgical method that can effectively repair the bone and/or tendon exposed wounds in fingers of children after electric burns. After wound healing, the scars are slight, the finger function is well recovered, and the parents of the children are highly satisfied with the curative effect of the children, which is worthy of clinical promotion.

[Analysis of clinical characteristics and risk factors of death in critical burn patients complicated with invasive fungal infection].

Objective: To investigate the clinical characteristics and risk factors of critical burn patients complicated with invasive fungal infection. Methods: A retrospective case series study was conducted. From January 2017 to December 2022, 88 critical burn patients combined with invasive fungal infection who met the inclusion criteria were admitted to Zhengzhou First People's Hospital, including 61 males and 27 females, aged 26-74 years. Data on invasive fungal infection sites and the detection of pathogens in patients were recorded. According to the survival outcome within 28 days after admission, the patients were divided into survival group (63 cases) and death group (25 cases). The following data of patients were compared between the two groups, including the basic data and injuries of patients at admission such as age, sex, body weight, total burn area, combination of inhalation injury, combination of hypertension and diabetes, acute physiology and chronic health status evaluation Ⅱ (APACHE Ⅱ) score, and admission time after burns, the levels of blood biochemical indexes within 24 h after admission such as white blood cell count, platelet count, red blood cell count, monocyte count, neutrophil count, lymphocyte count, alanine transaminase, aspartate transaminase, glucose, creatinine, urea nitrogen, D-dimer, galactomannan (GM), 1,3-ß-D glucan, and creatine kinase, the application of invasive procedures and vasoactive drugs during the treatment such as continuous renal replacement therapy, ventilator-assisted breathing, tracheotomy, deep vein catheterization, skin grafting >2 times, the levels of infection indicators on post admission day (PAD) 1, 3, 7, and 14 including C-reactive protein (CRP), procalcitonin, lactic acid, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), and the detection of pathogens in the process of treatment. Data were statistically analyzed with independent sample t test, analysis of variance for repeated measurement, chi-square test, Mann-Whitney U test, and Bonferroni correction. Multivariate logistic regression analysis was performed to screen the independent risk factors that affected death of critical burn patients complicated with invasive fungal infection. Results: The main sites of invasive fungal infection were the wounds (67 cases) and blood stream (46 cases), with Candida fungi (58 strains) as the main strain for fungi infection, and there were a total of 30 cases of infection with mixed pathogenic bacteria. Compared with those in survival group, the APACHE Ⅱ score, proportions of combination with inhalation injury and hypertension of patients in death group were significantly increased (t=2.11, with χ2 values of 6.26 and 9.48, respectively, P<0.05), while the other basic data and injury condition had no significant changes (P>0.05). Compared with those in survival group, the levels of D-dimer, GM, and 1,3-ß-D glucan of patients in death group were significantly increased within 24 h after admission (with t values of 2.42, 2.05, and 2.21, respectively, P<0.05), while the other blood biochemical indexes within 24 h after admission, as well as the proportions of applying invasive procedures and application of vasoactive drugs during the treatment process were not significantly changed (P>0.05). The levels of infection indicators of patients on PAD 1 and 3 were similar between the two groups (P>0.05). The procalcitonin level on PAD 7 and the levels of CRP, procalcitonin, lactic acid, IL-6, and TNF-α on PAD 14, as well as the proportion of infection with mixed pathogenic bacteria of patients in death group were significantly higher than those in survival group (with t values of 4.69, 3.89, 6.70, 6.14, 4.65, and 3.26, respectively, χ2=12.67, P<0.05). Multivariate logistic regression analysis showed that combination with inhalation injury, combination with hypertension, and infection with mixed pathogenic bacteria were independent risk factors for death of critical burn patients complicated with invasive fungal infection (with odds ratios of 5.98, 4.67, and 6.19, respectively, 95% confidence intervals of 1.42-15.39, 1.41-25.28, and 1.86-20.58, respectively, P<0.05). Conclusions: The main sites of infection in critical burn patients complicated with invasive fungal infection are the wounds and blood stream, with Candida fungi as the main strain for fungi infection, and a large proportion of infection with mixed pathogenic bacteria. The combined inhalation injury, combined hypertension, and infection with mixed pathogenic bacteria are the independent risk factors for the death of those patients.

[Effects of expanded frontal-parietal pedicled flap in reconstructing cervical scar contracture deformity in children after burns].

Objective: To explore the effects of expanded frontal-parietal pedicled flap in reconstructing cervical scar contracture deformity in children after burns. Methods: A retrospective observational study was conducted. From January 2015 to December 2020, 18 male children with cervical scar contracture deformity after burns who met the inclusion criteria were admitted to Zhengzhou First People's Hospital, aged 4 to 12 years, including 10 cases with degree Ⅱ cervical scar contracture deformity and 8 cases with degree Ⅲ scar contracture deformity, and were all reconstructed with expanded frontal-parietal pedicled flap. The surgery was performed in 3 stages. In the first stage, a cylindrical skin and soft tissue expander (hereinafter referred to as expander) with rated capacity of 300 to 500 mL was placed in the frontal-parietal region. The expansion time was 4 to 6 months with the total normal saline injection volume being 2.1 to 3.0 times of the rated capacity of expander. In the second stage, expander removal, scar excision, contracture release, and flap transfer were performed, with the flap areas of 18 cm×9 cm to 23 cm×13 cm and the secondary wound areas of 16 cm×8 cm to 21 cm×11 cm after scar excision and contracture release. After 3 to 4 weeks, in the third stage, the flap pedicle was cut off and restored. The rated volume of placed expander, total normal saline injection volume, type of vascular pedicle of flap, survival of flap and reconstruction of scar after the second stage surgery were recorded. The neck range of motion and cervico-mental angle were measured before surgery and one-year after surgery. The appearance of neck, occurrence of common complications in the donor and recipient sites of children, and satisfaction of children's families for treatment effects were followed up. Data were statistically analyzed with paired sample t test. Results: All the patients successfully completed the three stages of operation. The rated volume of implanted expander was 300 mL in 6 children, 400 mL in 9 children, and 500 mL in 3 children, with the volume of normal saline injection being 630 to 1 500 mL. The type of vascular pedicle of flap was double pedicle in 13 cases and was single pedicle in 5 cases. All the flaps in 17 children survived well, and the secondary wounds after neck scar excision and contracture release were all reconstructed in one procedure. In one case, the distal blood supply of the single pedicled flap was poor after the second stage surgery, with necrosis of about 2.5 cm in length. The distal necrotic tissue was removed on 10 days after the operation, and the wound was completely closed after the flap was repositioned. In the follow-up of 6 months to 3 years post operation, the cervical scar contracture deformity in 18 children was corrected without recurrence. The flap was not bloated, the texture was soft, and the appearances of chin and neck were good. The range of motion of cervical pre-buckling, extension, left flexion, and right flexion, and cervico-mental angle in one year after operation were improved compared with those before operation (with t values of 43.10, 22.64, 27.96, 20.59, and 88.42, respectively, P<0.01). The incision in the frontal donor site was located in the hairline, the scar was slight and concealed. No complication such as cranial depression was observed in expander placement site, and the children's families were satisfied with the result of reconstruction. Conclusions: Application of expanded frontal-parietal pedicled flap in reconstructing the cervical scar contracture deformity in children after burns can obviously improve the appearance and function of neck, with unlikely recurrence of postoperative scar contractures, thus it is an ideal method of reconstruction.

[Value of cerebral hypoxic-ischemic injury markers in the early diagnosis of sepsis associated encephalopathy in burn patients with sepsis].

Objective: To explore the value of cerebral hypoxic-ischemic injury markers in the early diagnosis of sepsis associated encephalopathy (SAE) in burn patients with sepsis. Methods: A retrospective case series study was conducted. From October 2018 to May 2021, 41 burn patients with sepsis who were admitted to Zhengzhou First People's Hospital met the inclusion criteria, including 23 males and 18 females, aged 18-65 (35±3) years. According to whether SAE occurred during hospitalization, the patients were divided into SAE group (21 cases) and non-SAE group (20 cases). The gender, age, deep partial-thickness burn area, full-thickness burn area, and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) scores of patients were compared between the two groups. The serum levels of central nervous system specific protein S100ß and neuron specific enolase (NSE) at 12, 24, and 48 h after sepsis diagnosis (hereinafter referred to as after diagnosis), the serum levels of interleukin-6 (IL-6), IL-10, tumor necrosis factor α (TNF-α), Tau protein, adrenocorticotropic hormone (ACTH), and cortisol at 12, 24, 48, 72, 120, and 168 h after diagnosis, and the mean blood flow velocity of middle cerebral artery (VmMCA), pulsatility index, and cerebral blood flow index (CBFi) on 1, 3, and 7 d after diagnosis of patients in the two groups were counted. Data were statistically analyzed with chi-square test, analysis of variance for repeated measurement, independent sample t test, and Bonferroni correction. The independent variables to predict the occurrence of SAE was screened by multi-factor logistic regression analysis. The receiver operating characteristic (ROC) curve was drawn for predicting the occurrence of SAE in burn patients with sepsis, and the area under the curve (AUC), the best threshold, and the sensitivity and specificity under the best threshold were calculated. Results: The gender, age, deep partial-thickness burn area, full-thickness burn area, and APACHE Ⅱ score of patients in the two groups were all similar (χ2=0.02, with t values of 0.71, 1.59, 0.91, and 1.07, respectively, P>0.05). At 12, 24, and 48 h after diagnosis, the serum levels of S100ß and NSE of patients in SAE group were all significantly higher than those in non-SAE group (with t values of 37.74, 77.84, 44.16, 22.51, 38.76, and 29.31, respectively, P<0.01). At 12, 24, 48, 72, 120, and 168 h after diagnosis, the serum levels of IL-10, Tau protein, and ACTH of patients in SAE group were all significantly higher than those in non-SAE group (with t values of 10.68, 13.50, 10.59, 8.09, 7.17, 4.71, 5.51, 3.20, 3.61, 3.58, 3.28, 4.21, 5.91, 5.66, 4.98, 4.69, 4.78, and 2.97, respectively, P<0.01). At 12, 24, 48, 72, and 120 h after diagnosis, the serum levels of IL-6 and TNF-α of patients in SAE group were all significantly higher than those in non-SAE group (with t values of 8.56, 7.32, 2.08, 2.53, 3.37, 4.44, 5.36, 5.35, 6.85, and 5.15, respectively, P<0.05 or P<0.01). At 12, 24, and 48 h after diagnosis, the serum level of cortisol of patients in SAE group was significantly higher than that in non-SAE group (with t values of 5.44, 5.46, and 3.55, respectively, P<0.01). On 1 d after diagnosis, the VmMCA and CBFi of patients in SAE group were significantly lower than those in non-SAE group (with t values of 2.94 and 2.67, respectively, P<0.05). On 1, 3, and 7 d after diagnosis, the pulsatile index of patients in SAE group was significantly higher than that in non-SAE group (with t values of 2.56, 3.20, and 3.12, respectively, P<0.05 or P<0.01). Serum IL-6 at 12 h after diagnosis, serum Tau protein at 24 h after diagnosis, serum ACTH at 24 h after diagnosis, and serum cortisol at 24 h after diagnosis were the independent risk factors for SAE complicated in burn patients with sepsis (with odds ratios of 2.42, 1.38, 4.29, and 4.19, 95% confidence interval of 1.76-3.82, 1.06-2.45, 1.37-6.68, and 3.32-8.79, respectively, P<0.01). For 41 burn patients with sepsis, the AUC of ROC of serum IL-6 at 12 h after diagnosis for predicting SAE was 0.92 (95% confidence interval was 0.84-1.00), the best threshold was 157 pg/mL, the sensitivity was 81%, and the specificity was 89%. The AUC of ROC of serum Tau protein at 24 h after diagnosis for predicting SAE was 0.92 (95% confidence interval was 0.82-1.00), the best threshold was 6.4 pg/mL, the sensitivity was 97%, and the specificity was 99%. The AUC of ROC of serum ACTH at 24 h after diagnosis for predicting SAE was 0.96 (95% confidence interval was 0.89-1.00), the best threshold was 14.7 pg/mL, the sensitivity was 90%, and the specificity was 94%. The AUC of ROC of serum cortisol at 24 h after diagnosis for predicting SAE was 0.93 (95% confidence interval was 0.86-1.00), the best threshold was 89 nmol/L, the sensitivity was 94%, and the specificity was 97%. Conclusions: Serum Tau protein, ACTH, and cortisol have high clinical diagnostic value for SAE complicated in burn patients with sepsis.

IGHG1 functions as an oncogene in tongue squamous cell carcinoma via JAK1/STAT5 signaling.

The article "IGHG1 functions as an oncogene in tongue squamous cell carcinoma via JAK1/STAT5 signaling, by Y.-L. Zheng, Y.-Y. Li, J.-F. Xie, H.-Q. Ma, published in Eur Rev Med Pharmacol Sci 2020; 24 (12): 6716-6725-DOI: 10.26355/eurrev_202006_21659-PMID: 32633362" has been withdrawn from the authors stating that "the experimental data in the article are wrong". The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/21659.

[Clinical effects of single pedicle transfer of expanded axial flap across the midline of the frontal-parietal region in reconstruction of large scar deformities in the face and neck].

Objective: To explore the clinical effects of single pedicle transfer of expanded axial flap across the midline of the frontal-parietal region in reconstruction of large scar deformities in the face and neck. Methods: From January 2016 to August 2019, 10 male patients, aged from 20 to 52 years with post-burn facial and cervical scar deformities, were admitted to the First People's Hospital of Zhengzhou, with the size of scar ranging from 15 cm×7 cm to 23 cm×11 cm. In the first stage, a cylindrical skin and soft tissue expander with rated capacity ranging from 400 to 600 mL was placed in the frontal-parietal region. Another cylindrical expander with rated capacity ranging from 50 to 100 mL was placed in the temporal region of the patient with scars in front of the ear and in cheek. The injection time was 3 to 5 months with the total injection volume being 1.5 to 2.5 times of the rated capacity of expander. In the second stage, the superficial temporal artery frontal branch and its branches were explored, the expander was removed, the scars in the face and neck were conducted resection and contracture relaxation, and the single pedicle transfer of expanded axial flap across the midline of the frontal-parietal region for reconstruction was performed. When the branches of the superficial temporal vessels were difficult to be detected by Doppler ultrasonic blood stream detector, the patient underwent computed tomography (CT) angiography and three-dimensional reconstruction. The donor site in frontal-parietal region was directly sutured, and the wound of the exposed donor site at the pedicle and temporal region was temporarily covered with scar skin. After the suture wound was healed and the hair in expanded flap grew out, hair removal and laser hair removal were performed. Three to four weeks after transplantation of expanded flap, the flap pedicle was cut off, restored, and trimmed in the third stage. The status about the completion of operation, the implantation of expander in the temporal region, CT angiography and three-dimensional reconstruction were recorded. The effective resection area of expanded flap, the length across the midline and the length of the pedicle, and the survival status of the expanded flap and complications after operation were observed. The appearance of donor and recipient sites, the scar recurrence, the appearance and function improvement of patients, and the satisfaction degree of patients were followed up. Results: All the 10 patients successfully completed three stages of operation, of which 6 patients had an auxiliary expander placement in the temporal region, and 5 patients underwent CT angiography and three-dimensional reconstruction. The effective resection area of expanded flap ranged from 18 cm×8 cm to 25 cm×13 cm. The distal end of the flap across the midline extended 4-6 cm to the opposite side, and the length of pedicle was 2-6 cm. All the expanded flaps of patients survived well after formation and transfer. The venous reflux disorder and obvious swelling occurred in 6 patients at the distal end of the flap after operation, and the blood supply recovered after acupuncture bloodletting, etc. Follow-up of 6 to 24 months showed that the color, texture, and thickness of the expanded flaps were similar to those of the facial skin, and no recurrence of scar was observed; the incision in the donor site of the frontal-parietal region was concealed, the hair growth of the temples and head was normal, and the reconstructed hairline was natural; compared with those before operation, the appearance, head-up, mouth-opening, and other functions of patients were significantly improved; the patients were satisfied with the effect of reconstruction. Conclusions: Clinical application of single pedicle transfer of expanded axial flap across the midline of the frontal-parietal region in reconstruction of large scar deformities in the face and neck can achieve a good appearance and function, and the donor site shows good shape, which enriches the application range of the trans-regional blood supply flap. It is a reliable method for reconstruction of large scar deformities in the face and neck.

IGHG1 functions as an oncogene in tongue squamous cell carcinoma via JAK1/STAT5 signaling.

OBJECTIVE: We explored the IgG1 heavy chain constant region (IGHG1) roles in tongue squamous cell carcinoma (TSCC) progression, as well as to probe the underlying mechanisms. PATIENTS AND METHODS: The expression patterns of IGHG1 in TSCC tissues and cell lines were tested by Western blotting, quantitative real-time PCR (RT-PCR) and immunohistochemistry (IHC) technologies. The relationship between IGHG1 expression level and the overall survival and clinicopathologic features of patients with TSCC were evaluated to assess the clinical value of IGHG1. The effects of IGHG1 on cell function were determined by Cell-Counting Kit-8 (CCK-8), clone formation, flow cytometry and in vivo tumor formation assays. RESULTS: The expression of IGHG1 in TSCC tissues and cell lines was significantly elevated at both mRNA and protein levels. IGHG1 expression levels closely related to T classification (p=0.008), clinical stage (p=0.011), and node metastasis (p=0.005) in TSCC patients. Upregulation of IGHG1 with lentivirus infection significantly increased Janus kinase 1 (JAK1) expression and the phosphorylation level of signal transducer and activator of transcription 5 (STAT5). In addition, IGHG1 overexpression markedly enhanced cell proliferation, clone formation and tumorigenesis and inhibited cell apoptosis, whereas these effects were abolished when JAK1 was downregulated in SCC15 and SCC4 TSCC cell lines. CONCLUSIONS: Collectively, this study reveals that IGHG1 functions as an oncogene in TSCC via activating JAK1/STAT5 signaling.

Circulating SHIP2 mRNA as a novel biomarker in the diagnosis and prognosis of gastric cancer.

OBJECTIVE: Previous studies showed the aberrant expression of Src homology 2-containing inositol 5-phosphatase 2 (SHIP2) in GC tissue. However, the exact role of circulating SHIP2 in GC remains unclear. The aim of this manuscript was to analyze potential diagnostic and prognostic value of circulating SHIP2 levels in GC. PATIENTS AND METHODS: Circulating SHIP2 expression was detected in the plasma of 156 GC patients and 60 healthy controls by qRT-PCR. The receiver operating characteristic (ROC) curves were plotted to explore the reliability of circulating SHIP2 in detecting GC. Survival curves were estimated using the Kaplan-Meier method, and differences between them were evaluated by the log-rank test. The influence of each variable on survival was examined by the Cox multivariate regression analysis. RESULTS: Our research showed that the expression levels of circulating SHIP2 in plasma of GC patients were lower than in healthy controls (p < 0.05). Decreased circulating SHIP2 mRNA expression was negatively correlated with clinical stage (p = 0.004), lymph node metastasis (p= 0.003) and distant metastasis (p = 0.025). ROC curve analysis showed that circulating SHIP2 may be a useful marker for discriminating cases from healthy controls. In addition, patients with low circulating SHIP2 mRNA level had poorer overall survival than those with high circulating SHIP2 mRNA level (p = 0.006). Moreover, multivariate analysis indicated that the level of circulating SHIP2 mRNA expression was an independent prognostic indicator (p = 0.005) for the survival of patients with GC. CONCLUSIONS: The present study indicated that decreased plasma SHIP2 mRNA level might be a novel diagnostic and prognostic biomarker for GC patients. This conclusion should be further assessed in randomized clinical trials.

Impact Factors and Attitudes Toward Organ Donation Among Transplantation Patients and Their Caregivers in China.

PURPOSE: This study's purpose was to investigate the attitudes toward organ donation among renal transplantation patients and their caregivers. In addition, we sought to explore the impact factors that affect their attitudes toward deceased organ donation. DESIGN AND METHODS: A self-administrated questionnaire was used, which consisted of two parts: 1) demographic data, and 2) transplantation and donation-related data. This study was conducted in three transplantation follow-up centers in three hospitals using a cross-sectional approach. SPSS 17.0 software was used to analysis descriptive and inferential statistics for data. The responses were analyzed using descriptive statistics and logistic regression analysis. RESULTS: We received 426 effective questionnaires. The renal transplantation patients' mean age was 40.84 years. Among these patients, 67.8% were willing to accept the organ transplantation surgery for their relatives, 67.4% were willing to donate a living kidney to a close relative, 62.7% were willing to donate organs after death, 53.5% were willing to register in the national organ donation system, and 51.4% were willing to sign the organ donation consent when facing their relatives becoming a potential organ donor. Age, marriage status, education level, understanding of transplantation procedures and understanding of donation procedures had statistical significance in the difference of the attitudes toward donate their organs after death (P < .05). CONCLUSIONS: Renal transplantation patients in our study are more willing to donate organs after death than their caregivers, but both their attitudes toward deceased donation were not very optimistic. There is a significant relationship between participants' willingness and knowledge of organ donation; patients with more understanding of the transplantation and donation procedure were more willing to donate organs after death. Affected by traditional values such as Confucianism, many people still cannot accept registering in the national organ donation system or sign the organ donation consent when facing their relatives becoming potential organ donors. CLINICAL RELEVANCE: There is a need to give adequate training regarding donation to increase donation rates. The government must provide education from the perspective of scientific knowledge to change the traditional views of the public, which may then increase the donation rate in China.

Attitude and Impact Factors Toward Organ Transplantation and Donation Among Transplantation Nurses in China.

BACKGROUND: Health workers' awareness and knowledge of transplantation medicine can improve people's sensitivity and reduce their degree of opposition to donations. The medical literature contains numerous examples of attitudes toward organ transplantation and donation aimed at university students or medical staff members, but rarely for transplantation nurses. OBJECTIVE: The purposes of the study were to investigate the attitudes toward organ transplantation and donation among transplantation nurses and to explore the impact factors. METHODS: The study was conducted in 37 transplantation surgery wards in 22 hospitals using cross-sectional approach. SPSS (International Business Machines Corporation, Armonk, New York, USA) 7.0 software was used to analysis descriptive and inferential statistics for data. RESULTS: Five hundred thirty-six effective questionnaires were received and the effective rate was 89.33%. Nurses' mean age was 28.40 years with a mean service length of 6.54 years. Among these nurses, 66.6% and 78.0% were willing to accept organ transplantation surgery for themselves and their relatives, respectively. Of these nurses, 33.4% would donate their organs after death; whereas 39.9% were uncertain. Only 38.2% were willing to register in the national organ donation system. Of these nurses, 28.2% were willing to sign the organ donation consent forms when their relatives became potential organ donors, and 45.7% were uncertain. Eight independent variables that affected nurses' attitudes toward donating their organs from most to least significant were: ratio of nurse to bed, title, employment form, age, length of service, position, monthly income, and the highest educational degree earned. Pearson correlation analysis showed a significant correlation among nurses' attitudes toward organ transplantation, organ donation, and online registration. CONCLUSION: The attitude toward donation and transplantation in the hospitals was not too optimistic, and an improvement in the training regarding transplantation and donation among nurses in China is necessary. Nurses are an important group who generate opinion in the patient population, and their negative attitudes can have a significant negative impact on society's attitudes toward organ donation.

Regulation of tumor necrosis factor alpha-mediated apoptosis of rheumatoid arthritis synovial fibroblasts by the protein kinase Akt.

OBJECTIVE: To determine if tumor necrosis factor alpha (TNFalpha)-driven proliferation of rheumatoid arthritis synovial fibroblasts (RASF) is associated with up-regulation of the activity of serine/threonine kinase B/Akt and with survival of RASF. METHODS: Staining of phosphorylated Akt was done using anti-phosphorylated Thr308 Akt antibody. Levels of phosphorylated Akt were analyzed by Western blot and Akt activity was analyzed using a kinase assay. TUNEL staining was used to analyze the cytotoxicity of TNFalpha treatment or TNFalpha combined with either the Akt activity inhibitor wortmannin, an adenovirus expressing dominant-negative mutant (AdAkt-DN), or an adenovirus expressing phosphatase and tensin homolog deleted on chromosome 10 (AdPTEN). RESULTS: The levels of phosphorylated Akt were higher in RASF than in osteoarthritis synovial fibroblasts (OASF), as demonstrated by immunohistochemical staining, immunoblot analysis, and an Akt kinase assay. The levels of phosphorylated Akt and Akt kinase activity were increased by stimulation of primary RASF with TNFalpha (10 ng/ml). Treatment of RASF with the phosphatidylinositol 3-kinase inhibitor wortmannin (50 nM) plus TNFalpha resulted in apoptosis of 60 +/- 8% (mean +/- SEM) of RASF within 24 hours. This proapoptosis effect was specific for Akt, since equivalent levels of apoptosis were observed upon TNFalpha treatment of RASF transfected with AdAkt-DN and with AdPTEN, which opposes the action of Akt. CONCLUSION: These results indicate that phosphorylated Akt acts as a survival signal in RASF and contributes to the stimulatory effect of TNFalpha on these cells by inhibiting the apoptosis response. This effect was not observed in OASF and may reflect the pathophysiologic changes associated with the proliferating synovium in rheumatoid arthritis.

Recombinant adenovirus expressing adeno-associated virus cap and rep proteins supports production of high-titer recombinant adeno-associated virus.

It has been difficult to produce a chimeric vector containing both Ad and AAV rep and cap, and to grow such chimeric vectors in 293 cells. By recombination in vitro in a bacterial host, we were able to produce recombinant plasmid AdAAV (pAdAAVrep-cap), which could be used to generate recombinant AdAAV (rAdAAVrep-cap) after transfection into 293 cells. A recombinant adenovirus, rAdAAVGFP, in which the green fluorescent protein (GFP) gene is flanked by the AAV terminal repeats cloned into the E1-deleted site of Ad was also generated. Co-infection of rAdAAVrep-cap together with rAdAAVGFP into 293 cells resulted in production of high titers of rAAV expressing GFP. It was noted that the titer of rAdAAVrep-cap was lower than the titer of control AdCMVLacZ. The lower titer of rAdAAvrep-cap was associated with expression of Rep protein. Non-homologous recombination occurs after high passage and results in deletions within the AAV rep genes. These results indicate that (1) rAdAAVrep-cap can be produced; (2) rAdAAVrep-cap + rAdAAVGFP is a convenient and efficient way to transfect 293 cells to grow high titer rAAV; and (3) frozen stock is required to avoid propagation of rep-deleted pAdAAVrep-cap.

A complex element regulates IFN-gamma-stimulated monocyte chemoattractant protein-1 gene transcription.

Monocyte chemoattractant protein-1 (MCP-1) is induced in chronic osseous inflammation, and is temporally and spatially correlated with monocyte recruitment. We investigated the mechanism of MCP-1 regulation in a human osteoblastic cell line in response to IFN-gamma, a potent mediator of the immune inflammatory response. Nuclear run-on and stability studies demonstrated that IFN-gamma stimulated MCP-1 transcription and did not enhance mRNA stabilization. Using MCP-1 promoter/reporter gene constructs, we determined that IFN-gamma-enhanced MCP-1 transcription is regulated by a 29-bp element located at -227 relative to the ATG start codon. This element contains a 13-bp CT-rich sequence (GCTTCCCTTTCCT) adjacent to a IFN-gamma activation site (GAS). Since deletion of the CT sequence enhanced both the magnitude and duration of IFN-gamma-stimulated, GAS-mediated transcription, we have termed it the IFN response-inhibitory sequence (IRIS). The combined IRIS/GAS sequence is highly conserved in mouse, rat, and bovine MCP-1 genes. In gel-shift assays, nuclear extracts from IFN-gamma-stimulated osteoblastic cells formed two specific inducible bands with labeled IRIS/GAS DNA. Both bands were supershifted by anti-STAT1 Abs, but not by Abs to STAT2, p48(ISGF-3y), IFN-regulatory factor-1, or IFN-regulatory factor-2. Formation of one of the bands required the presence of the IRIS moiety. IRIS/GAS DNA also formed a number of specific complexes with constitutively expressed factors, none of which were affected by the above Abs. These studies establish a mechanism for IFN-gamma-stimulated MCP-1 expression and identify a complex element that regulates MCP-1 gene transcription.

Pro-inflammatory cytokines downregulate platelet derived growth factor-alpha receptor gene expression in human osteoblastic cells.

Platelet derived growth factor (PDGF) is thought to play a significant role in bone repair and regeneration. We previously demonstrated that PDGF-AA binding can be modulated by interleukin-1 (IL-1). We now report that TNF-alpha significantly reduces PDGF-AA binding by decreasing the number of PDGF-alpha receptor subunits on the surface of normal human osteoblastic cells. This inhibition is likely due to a decrease in synthesis of PDGF-alpha receptors since TNF-alpha causes a relatively rapid decrease in PDGF-alpha receptor mRNA levels as determined by Northern blot analysis. The physiologic importance of this inhibition is demonstrated by a TNF-alpha induced decrease in PDGF-AA stimulated tyrosine kinase activity. When saturating concentrations of TNF-alpha were used, the addition of IL-1 further inhibited PDGF-AA binding and further decreased surface expression of PDGF-alpha receptors. In contrast, other mediators such as IL-6, PTH, 1,25(OH)2 vit D3, hydrocortisone, PGE2, bFGF, and IGF-1 had no effect. These results suggest that binding to the PDGF-alpha receptor is decreased by the strong pro-inflammatory cytokines such as IL-1 beta and TNF-alpha rather than as a general response to mediators important in bone resorption or bone formation. TNF-alpha and IL-1 are often co-expressed during destructive inflammatory processes. Thus, TNF-alpha and IL-1 may work in concert to limit the response of osteoblastic cells to PDGF-AA during periods of osseous inflammation.

IL-1 down-regulates platelet-derived growth factor-alpha receptor gene expression at the transcriptional level in human osteoblastic cells.

Regulation of the platelet-derived growth factor (PDGF)-alpha receptor is thought to play an important role in pathophysiologic processes. Previously, we have reported that IL-1 has the potential to regulate PDGF-induced biologic activity in both normal human osteoblastic cells and the human osteoblastic cell line, MG-63, by decreasing the expression of PDGF-alpha receptor mRNA. In the present studies, we analyzed the effects of IL-1 on transcription rates and the stability of PDGF-alpha receptor mRNA in MG-63 cells. The data indicate that the t1/2 of PDGF-alpha receptor mRNA is approximately 3.3 h after incubation with the RNA II polymerase transcription inhibitor 5,6-dichloro-1 beta-D-ribofuranosylbenzimidazole (DRB). Approximately the same t1/2 (3.1 h) was obtained when osteoblastic cells were incubated with IL-1. The t1/2 for PDGF-alpha receptor mRNA for cells incubated with both IL-1 and DRB was 3 h. This finding suggests that the levels of PDGF-alpha receptor mRNA transcripts are not regulated by post-transcriptional mechanisms. Results of nuclear run-on analysis were consistent with this conclusion, demonstrating that IL-1 modulates PDGF-alpha receptor gene expression at the transcriptional level. Surprisingly, incubation of cells with cycloheximide also caused down-regulation of PDGF-alpha receptor mRNA, which suggests that synthesis of a labile factor is necessary for constitutive expression. The functional consequence of down-regulation of PDGF-alpha receptors by IL-1 was also assessed. By using chemotaxis assays, we demonstrated that IL-1 significantly inhibited PDGF-AA-mediated migration in human MG-63 osteoblastic sarcoma cells.

IL-1 and transforming growth factor-beta inhibit platelet-derived growth factor-AA binding to osteoblastic cells by reducing platelet-derived growth factor-alpha receptor expression.

Platelet-derived growth factor (PDGF) is thought to play a significant role in bone repair and regeneration. We previously demonstrated that PDGF-AA-induced chemotaxis and proliferation can be modulated by IL-1. We now report that IL-1 and transforming growth factor-beta (TGF-beta) significantly decrease the number of PDGF-AA binding sites in both normal and tumor-derived human osteoblastic cells, whereas PDGF-BB binding is minimally affected. The affinity of PDGF-AA binding remains unchanged in the presence of IL-1, but is slightly reduced by TGF-beta as demonstrated by Scatchard analysis. We also showed that tyrosyl kinase phosphorylation after PDGF-AA binding is decreased in the presence of both IL-1 and TGF-beta. Northern blot analysis indicates that both IL-1 and TGF-beta decrease the expression of PDGF-alpha receptor mRNA. These results suggest that IL-1 and TGF-beta have the potential to regulate PDGF-AA-induced biologic activity in normal human osteoblastic cells and in human osteoblastic sarcoma cells by decreasing the levels of the PDGF-alpha receptor.

In vitro and animal studies of the role of viruses in oral carcinogenesis.

The linkage of herpes simplex virus (HSV) and human papillomavirus (HPV) to the development of oral cancer has been studied. In spite of the presence of viral nucleic acids in some human oral cancer specimens, HSV alone is not carcinogenic in animals: repeated viral inoculation to mouse or hamster oral mucosa fails to produce tumours or histopathological evidence of malignancy. However, HSV demonstrates co-carcinogenicity in vivo: viral inoculation significantly enhances the oncogenic capacity of chemical carcinogens in the oral cavity of mice and hamsters. Though the detailed mechanisms of HSV cocarcinogenicity are unknown, HSV promotes the chemical carcinogen-induced activation of certain cellular proto-oncogenes and inactivation of p53 tumour suppressor gene. Human papillomaviruses type 16 (HPV-16) and 18 (HPV-18) demonstrate oncogenicity by transforming normal human oral keratinocytes in vitro. While normal cells exhibit a limited life-span, cells transformed by these viruses show immortality and altered morphology in comparison with their normal counterparts. The HPV-immortalised cells contain multiple copies of intact viral genome integrated into cellular chromosomes. These cells also express several viral-specific mRNAs including viral E6/E7 mRNAs. Notably, these cells contain low levels of p53 protein and overexpressed cellular myc proto-oncogene compared to their normal counterpart; however, the immortilised cell lines are non-tumorigenic in nude mice.

[The blocking effect of topically subepithelial injection of thioproline and vitamin A acid in experimental oral premalignant].

The purpose of this paper is to observe the blocking effect of topically subepithelial injected drug and Vit A acid painting on chemically induced oral precancerous lesion and to prove, on a certain extent, the hypothesis that subepithelial connective tissue could exert great influence on the differentiation of the epithelium. A total of 49 syrian hamster was used as experimental animal. Both buccal pouches of all animals were painted thrice weekly with 0.5% DMBA in acetone for 6 weeks. Then, they were divided into two groups: Control group and Experimental group. In the latter group, 70.8% precancerous lesions turned into normal epithelial tissue, whereas those untreated animals developed carcinoma by 62%.

[Clinical significance of serum ferritin and acute phase reactant proteins levels in patients with cervical cancer].

Serum ferritin (SF) levels from 162 patients with cervical cancer and their serum alpha 1-acid glycoprotein (alpha 1-AGP). alpha 1-antitrypsin (alpha 1-AT. Transferrin (Tf) in most patients were determined. The result showed that concentration of SF, alpha 1-AGP, alpha 1-AT were significantly higher, while TF significantly lower in cervical carcinoma patients during active period than from patients with benign tumors and normal persons. The levels of SF. alpha 1-AGP. alpha 1-AT and Tf were significantly increased during the remission period. The positive rate of SF, alpha 1-AGP and Tf in cervical cancer patients during active period was significantly higher than that of alpha 1-AT. Serial determinations of SF, alpha 1-AGP and. Tf may be helpful in the monitoring of disease development and early detection of recurrence and metastases.