[Correlation analysis of dynamic enhanced energy spectrum CT parameters with Ki-67 high expression in hepatocellular carcinoma].

Objective: To investigate the correlation between dynamic enhanced energy spectral CT parameters and Ki-67 high expression in hepatocellular carcinoma (HCC). Methods: A cross-sectional study. This retrospective case-control study analyzed the clinical data of 101 patients with pathologically confirmed HCC in Xiamen Hospital of Traditional Chinese Medicine and Zhongshan Hospital of Xiamen University from December 2017 to March 2023. These 101 patients included 84 males and 17 females, and the age[M(Q1, Q3)] was 59.0(49.0,66.0)years. These patients were divided into two groups according to the immunohistochemical Ki-67 expression levels in tumor tissues: the high expression group (Ki-67%>20%, n=59) and the low expression group (Ki-67%≤20%, n=42).CT values on 70 keV and 140 keV monochromatic energy images (HU70 keV-a, HU140 keV-a, HU70 keV-p, HU140 keV-p) and water density (Dwater-a, Dwater-p) were measured in arterial phase and portal vein phase, and the difference of HU70 keV, HU140 keV, Dwater values between portal vein and arterial phase (ΔHU70 keV, ΔHU140 keV, ΔDwater), as well as ratio of HU70 keV, HU140 keV, Dwatervalues between portal vein and arterial phase (HU70 keVratio, HU140 keVratio, Dwaterratio) were calculated. Spearman correlation analysis was used to analyze the correlation between the CT spectral parameters and Ki-67%. Multivariate logistic regression model was used to determine the factors associated with high expression of Ki-67. The receiver operating characteristics (ROC) curves were used to indicate the efficacy of dynamic enhanced spectral CT in evaluating Ki-67 high expression in HCC. Results: The high Ki-67 expression group revealed higher alpha fetal protein levels, larger tumor diameter and more irregular tumor shape compared with the low Ki-67 expression group,and the differences were statistically significant (all P<0.05). Spearman correlation analysis showed that the HU140 keV-p, Dwater-p, HU70 keV ratio, HU140 keV ratio, Dwater ratio, ΔHU70 keV, ΔHU140 keV, ΔDwater were positively correlated with Ki-67 positivity rate (r:0.31-0.50, all P<0.05). The spectral CT parameters (HU70 keV-p, HU140 keV-p, Dwater-p, HU70 keV ratio, HU140 keV ratio, Dwater ratio, ΔHU70 keV, ΔHU140 keV, ΔDwater) in high Ki-67 expression group were significantly higher than those in low Ki-67 expression group (all P<0.05). Multivariate logistic regression model shows that Dwater-p(OR=1.16, 95%CI: 1.05-1.29, P=0.005), ΔHU140 keV(OR=1.39, 95%CI: 1.20-1.62, P<0.001) and irregular tumor morphology (OR=5.25, 95%CI: 1.61-17.12, P=0.006) were correlative factors for high Ki-67 expression. The HU140 keV ratio and ΔHU140 keV alone evaluated the highest AUC of high Ki-67 high expression in HCC, which were 0.82 (95%CI: 0.74-0.90), the sensitivity were 61.0%, and the specificity were 88.1% and 85.7%. The combined analysis of Dwater-p, ΔHU140 keV and irregular tumor morphology had an increased AUC of 0.88 (95%CI: 0.81-0.95) in assessment high Ki-67 expression, with the sensitivity of 84.7% and the specificity of 78.6%. Conclusions: Dynamic enhanced spectral CT parameters were positively correlated with the Ki-67 expression in HCC. Spectral CT provides a non-invasive method to evaluate the proliferation status of HCC cells, and the efficiency could be improved by multi-parameter analysis combining spectral CT parameters and morphologic features.

[Research progress on the characteristics of artemisia pollen allergens and related pollinosis].

In recent years, the morbidity of pollinosis has been increasing year by year. Anemophilous flower pollen is the most important allergen causing pollinosis, among which artemisia pollen is one of the most common airborne allergens. In this paper, based on the immune biology characteristics of major sensitization protein components of artemisia pollen, and from the perspective of immunology, the main pathogenic mechanism of action and clinical characteristics of artemisia pollen are elaborated to provide the reference basis for the development of accurate and effective artemisia pollen disease prevention and control strategy, hoping to provide patients with scientific and effective prevention and control suggestions.

[Study on the relationship between ABCB1 gene polymorphism and hemorrhage risk after thrombolysis of cerebral ischemic stroke in Shangqiu area].

Using a cross-sectional study, 246 patients with hemorrhage and transformation after cerebral ischemic stroke(CIS) thrombolysis who were admitted to Shangqiu First People's Hospital, Shangqiu Municipal Hospital, and Shangqiu Liangyuan Traditional Chinese Medicine Hospital from March 2018 to May 2020 were selected as the observation group, 246 patients with no hemorrhage transformation after CIS thrombolysis during the same period were selected as the control group with a ratio of 1∶1. Polymerase chain reaction and pyrosequencing methods were used to detect the single nucleotide polymorphisms of the two groups of ABCB1 genes. The frequency distribution of each genotype of the two groups of ABCB1 gene polymorphism sites was counted. The conditional logistic regression equation was used to analyze the CIS after thrombolysis. Related influencing factors of hemorrhage transformation, and compare the single nucleotide polymorphisms of ABCB1 gene in patients with different prognosis in the observation group. The results showed that the CC genotype frequency of rs1045642 in the observation group was 34.55% higher than that of the control group 25.02%, the CT genotype frequency was 12.20%, and the TT genotype frequency 3.25% was lower than that of the control group 14.63% and 9.35% (χ2=21.527, P<0.05); GG genotype frequency at rs2032582 locus in observation group was 17.89%, GT genotype frequency 21.54% was lower than control group 37.60%, 93.96%, TT genotype frequency 10.57% higher than control group 2.44%, the difference was statistically significant (χ2=80.427, P<0.05); TT genotype at rs1045642 is a protective factor for hemorrhage transformation, and TT genotype at rs2032582 is a risk factor for hemorrhage transformation (OR=2.903, P<0.05). The risk of bleeding after thrombolysis in CIS patients in Shangqiu area may be related to the TT genotype at the ABCB1 rs1045642 locus and the TT genotype at the rs2032582 locus.

[Analysis of clinical characteristics, treatment and survival of elderly patients with large diffuse B-cell lymphoma].

Objective: To analyze the clinical feature,treatment and survival outcome of elderly patients older than 80 years with large diffuse B-cell lymphoma. Methods: A total of 46 patients aged over 80 years with large diffuse B-cell lymphoma who were treated in Third Hospital of Peking University during the period from 2002 to 2018 were retrospectively analyzed, and the clinical features, laboratory data, survival and prognostic factors were included in Kaplan-Meier and prognostic analysis. Results: Patients older than 80 years old accounted for 15.7% (46/293) in all elderly patients, and the median age was 83 years old. There were 78.3% (36/46)patients who belonged to stage Ⅲ or Ⅳ, 63%(29/46) who had more than two extranodal organ involvement, and the higher proliferation index(Ki-67≥80%) was present in 53.7%(22/41) patients. Immunohistochemistry showed that 37% patients in 27 cases were double-expressed DLBCL. With a median follow-up of 25 months, the overall response rate (ORR) for the whole group was 63.0%, the complete response (CR) rate was 36.4%, the 2, 3-year progression-free survival (PFS) rate was 49.9% and 41.7%, the 2, 3-year overall survival (OS) rate was 54.6% and 43.6% respectively. The ORR for patients who received anthracycline-based therapies and non-anthracycline-based therapies were 81.8% and 55.0%, and the 3-year OS rate were 50.0% and 39.0%, respectively, but the difference was not statistically significant (P>0.05). 45.5% patients had hematologic toxicity of Grade Ⅲ or above, and 56.8% patients experienced infections during the treatment. Among the patients who died, the treatment-related mortality rate in group with high score of Charlson comorbidity index(CCI) was higher (43.8% vs 16.7%, P=0.03) . The National Comprehensive Cancer Network International Prognostic Index (NCCN-IPI) score, nodal involvement area ≥3, 6 cycles of chemotherapy, CCI score, initial treatment outcome and refractory-relapsed were predictive of overall survival. Multivariate analysis indicated the CCI score (HR=6.463, P=0.008) and initial treatment outcome (HR=0.086, P=0.001) were independent prognostic risk factors. Conclusions: The clinical and pathological features of patients older than 80 years were highly aggressive with poor chemotherapy tolerance and high adverse reaction rate. Anthracycline-based therapies may be less important in the treatment of DLBCL patients aged over 80 years. Patients with high CCI score have higher treatment-related mortality and CCI can help identify elderly patients who are suitable for larger chemotherapy dose.

Urban-rural disparity of preventive healthcare utilisation among children under the universal health insurance coverage in Taiwan: a national birth cohort analysis.

OBJECTIVE: In the context of universal health insurance coverage, this study aimed to determine whether urban-rural inequality still exists in preventive health care (PHC) amongst children in Taiwan. STUDY DESIGN: Prospective cohort study. METHODS: A total of 184,117 mothers and their children born in 2009 were identified as the study cohort. The number of children born in urban, satellite and rural areas was 40,176, 57,565 and 86,805, respectively. All children were followed for 7 years, before which a total of seven times PHC were provided by Taiwan's National Health Insurance (NHI) programme. Ordinal logistic regression models were used to associate urbanisation level with the frequency of PHC utilisation. Stratified analyses were further performed in accordance with the children's birth weight and the mothers' birthplace. RESULTS: Children from satellite areas had higher utilisation for the first four scheduled PHC visits. Children living in urban areas received more PHC for the fifth and sixth scheduled visits. Compared with those from rural areas, children in satellite areas exhibited a small but significant increase in odds in PHC utilisation, with a covariate-adjusted odds ratio (aOR) of 1.04 and 95% confidence interval (CI) of 1.02-1.06. By contrast, no significant difference was observed between rural and urban areas (aOR = 1.01). Further stratified analyses suggest more evident urban-rural difference in PHC utilisation amongst children with low birth weight and foreign-born mothers. CONCLUSIONS: Given a universal health insurance coverage and embedded mechanisms in increasing the availability of healthcare resources in Taiwan, a slight urban-rural difference is observed in PHC utilisation amongst children. Hence, sociodemographic inequality in utilisation of PHC still exists. This issue should be addressed through policy intervention.

2Hz-electroacupuncture attenuates heroin-seeking behaviors via adjusts CB1-Rs and CB2-Rs expression in relapse-relevant brain regions of heroin self-administration rats.

Opiate addiction has a high rate of relapse. The accumulating evidence shows that electroacupuncture (EA) may be effective for the treatment of opiate relapse. However, the change of expression of CB1-Rs and CB2-Rs involve in 2Hz EA anti-relapse pathway is still unclear. To explore the changes of expression of CB1-Rs and CB2-Rs, heroin self-administration (SA) model rats were adopted and treated using 2Hz EA. The expressions of CB1-Rs and CB2-Rs were observed using immunohistochemistry method. The results showed that, compared with the control group, active pokes in the heroin-addicted group increased, while the active pokes decreased significantly in 2Hz EA group compared with heroin-addicted group. Correspondingly, the expression of CB1-Rs in prefrontal cortex (PFC), hippocampus (Hip), nucleus accumbens (NAc) and ventral tegmental area (VTA) all increased significantly while the expression of CB2-Rs in those relapse-relevant brain regions decreased obviously in heroin-addicted group when compared with the control group. In addition, the expression of CB1-Rs obviously decreased in the 2Hz EA group while the expression of CB2-Rs in those relapse-relevant brain regions increased significantly when compared with the heroin-addicted group. It indicated that 2Hz EA could attenuate the heroin-evoked seeking behaviors effectively. The anti-relapse effects of 2Hz EA might be related to the decrease of CB1-Rs and increase of CB2-Rs expression in relapse-relevant brain regions of heroin SA rats.

[Hot research topics on cardiovascular diseases in occupational population: a bibliometric analysis].

Objective: To investigate the literature characteristics and research topics on cardiovascular diseases in the occupational population quantitatively via a bibliometric analysis, and to provide a reference for the selection of research directions. Methods: A search strategy was developed according to the words in Medical Subject Heading (MeSH) , and PubMed database was searched for articles on cardiovascular diseases in the occupational population published from 2006 to 2015. The information of published year, authors, journals, and MeSH words was extracted, frequently used MeSH words were screened out, and a cluster analysis was performed for frequently used MeSH words. Results: A total of 1 272 articles were found, and about 150 articles were published annually from 2011 to 2015. Most articles were published in Occupational and Environmental Medicine, and the author Tohr Nilsson had the highest number of published articles. The influencing factors mainly included occupational mental stress, smoking, and working system, and the health outcomes attracting the most attention were hypertension, hand-arm vibration, ischemic heart disease, and myocardial infarction. The articles were clustered into 5 types, and 5 hot topics were summarized. Conclusion: The bibliometric analysis of cardiovascular diseases in occupational population shows that hypertension and hand-arm vibration are hot research topics, which can provide a reference to researchers.

A functional single-nucleotide polymorphism in the ERCC1 gene alters the efficacy of narrowband ultraviolet B therapy in patients with active vitiligo in a Chinese population.

BACKGROUND: T lymphocytes have been shown to cause the destruction of melanocytes in vitiligo pathogenesis. Narrowband ultraviolet B (NB-UVB), as an effective therapeutic strategy in vitiligo, can lead to the formation of DNA photoproducts such as cyclobutane pyrimidine dimers (CPDs) in perilesional lymphocytes and thus induce skin immunosuppression. The repair of DNA photoproducts is performed mainly through the nucleotide excision repair (NER) pathway. We hypothesized that single-nucleotide polymorphisms (SNPs) in NER genes might influence the repair capacity of CPDs and thus contribute to variations in phototherapy efficiency. OBJECTIVES: To detect genetic polymorphisms in NER genes and their relationship with the efficacy of NB-UVB therapy in patients with active vitiligo. METHODS: We investigated the association of NER SNPs (XPA A23G, XPC Ci11A, XPC C2919A and ERCC1 C118T) with phototherapy efficacy in 86 patients with vitiligo who received NB-UVB treatment. Furthermore, we examined the impact of ERCC1 C118T on the apoptosis of T lymphocytes and CPD accumulation after NB-UVB irradiation. RESULTS: We found that patients with vitiligo with the ERCC1 codon 118 CC genotype showed better efficacy after NB-UVB irradiation than those with the ERCC1 118 TT and CT genotypes, whereas no such association was documented among the genotypes of XPA A23G, XPC Ci11A or XPC C2919A. Additionally, the apoptosis rates and CPD levels of lymphocytes after NB-UVB irradiation in patients with the ERCC1 118 CC genotype were significantly higher than those in patients with the ERCC1 118 TT and CT genotypes. CONCLUSIONS: The ERCC1 118 CC genotype confers better efficacy of NB-UVB therapy in patients with active vitiligo.

Identification of novel kinase inhibitors by targeting a kinase-related apoptotic protein-protein interaction network in HeLa cells.

OBJECTIVES: Protein kinases orchestrate activation of signalling cascades in response to extra- and intracellular stimuli for regulation of cell proliferation. They are directly involved in a variety of diseases, particularly cancers. Systems biology approaches have become increasingly important in understanding regulatory frameworks in cancer, and thus may facilitate future anti-cancer discoveries. Moreover, it has been suggested and confirmed that high-throughput virtual screening provides a novel, effective way to reveal small molecule protein kinase inhibitors. Accordingly, we aimed to identify kinase targets and novel kinase inhibitors. MATERIALS AND METHODS: A series of bioinformatics methods, such as network construction, molecular docking and microarray analyses were performed. RESULTS: In this study, we computationally constructed the appropriate global human protein-protein interaction network with data from online databases, and then modified it into a kinase-related apoptotic protein-protein interaction network. Subsequently, we identified several kinases as potential drug targets according to their differential expression observed by microarray analyses. Then, we predicted relevant microRNAs, which could target the above-mentioned kinases. Ultimately, we virtually screened a number of small molecule natural products from Traditional Chinese Medicine (TCM)@Taiwan database and identified a number of compounds that are able to target polo-like kinase 1, cyclin-dependent kinase 1 and cyclin-dependent kinase 2 in HeLa cervical carcinoma cells. CONCLUSIONS: Taken together, all these findings might hopefully facilitate discovery of new kinase inhibitors that could be promising candidates for anti-cancer drug development.

Molecular mechanisms of Lycoris aurea agglutinin-induced apoptosis and G2 /M cell cycle arrest in human lung adenocarcinoma A549 cells, both in vitro and in vivo.

OBJECTIVES: Lycoris is aurea agglutinin (LAA) has attracted rising attention due to its remarkable bioactivities. Here, we aimed at investigating its anti-tumor activities. MATERIAL AND METHODS: In vitro methods including MTT, cellular morphology observation, FCM and immunoblotting were performed. In vivo methods like detection of tumor volume, body weight and survival ratio, as well as TUNEL staining were performed. RESULTS AND CONCLUSION: LAA triggers G2 /M phase cell cycle arrest via up-regulating p21expression as well as down-regulating cdk-1cyclinA singling pathway, and induces apoptotic cell death through inhibiting PI3K-Akt survival pathway in human lung adenocarcinoma A549 cells. While LAA has no significant cytotoxic effect toward normal human embryonic lung fibroblast HELF cells, and moreover, LAA could amplify the antineoplastic effects of cisplatin toward A549 cells. Lastly LAA also bears anti-cancer and apoptosis-inducing effects in vivo, and it could decrease the volume and weight of subcutaneous tumor mass obviously as well as expand lifespan of mice. These findings may provide a new perspective for elucidating the complicated molecular mechanisms of LAA-induced cancer cell growth-inhibition and death, providing a new opportunity of LAA as a potential candidate anti-neoplastic drug for future cancer therapeutics.

Mississippi River flood of 2008: observations of a large freshwater diversion on physical, chemical, and biological characteristics of a shallow estuarine lake.

High nitrogen (N) loading to coastal aquatic systems can be expressed as increased algal production and subsequent low dissolved oxygen. In April, 2008, predictions for extreme flood stage for the Lower Mississippi River triggered the opening of the Bonnet Carré Spillway, a major release valve for the river. The spillway diverted approximately 8 km3 of water over one month of operation into Lake Pontchartrain with a concomitant 10000 t of NO3-N. Satellite imagery, physical, water quality, and chlorophyll a (chl a) measurements show that the Mississippi River plume mixed with < 40% of the lake during this time, and much of the nutrient load was transported to the coastal ocean. Nitrate, dissolved reactive phosphorus (P), and dissolved silica (Si) concentrations were 4.8, 5.0, and 3.2 times higher, respectively, within the river plume when compared with those of the lake water. Despite the high nutrient concentrations within the river plume, phytoplankton biomass, evidenced by chl a concentrations, was low. Much of the nutrient load appeared to bypass the lake and was transported to the coastal ocean during the opening of the diversion. The potential removal of a total of 7.6% of the N load from the Mississippi River during the one month of flood level flow may have been a contributing factor in the lower than predicted hypoxia zone off the Louisiana coast during the summer of 2008.

Identification of stage-specifically expressed genes of Trichinella spiralis by suppression subtractive hybridization.

SUMMARYNewborn larvae (NBL) and adult (Ad) stage-specifically expressed genes or members of gene families of Trichinella spiralis were identified by suppression subtractive hybridization (SSH). Six cDNA clones were identified as NBL stage-specific, including 1 member of the T. spiralis gene family encoding glutamic acid-rich proteins, 2 clones encoding novel serine proteases, 2 closely related clones encoding proteins that are members of a deoxyribonuclease II (DNase II)-like family and 1 clone with no similarity to known genes. Four stage-specific clones encoding homologues of retinoid X receptor, caveolin, C2H2 type zinc finger protein and a putative protein with no homology to known sequences were obtained from 3-day-old adult worms. One gene specifically up-regulated in the 5-day-old adult worms encoding a putative cuticle collagen was also identified.

Hyperthermia-regulated immunogene therapy.

One of the key milestones that must be reached before gene therapy becomes feasible for clinical cancer treatment is to be able to regulate therapeutic gene expression. This is true for most current cancer gene therapy approaches, since the majority of therapeutic genes are toxic to both tumour and normal tissues. Among the wide array of strategies available for regulating gene expression, hyperthermia represents a unique approach. Hyperthermic regulation of gene therapy is feasible because of the widely conserved heat shock response, which allows therapeutic gene expression to be elevated to thousands of fold higher than background when temperature reaches 3-5 degrees C over physiological temperature (37 degrees C). In addition, because of the long history of experimental research on the use of hyperthermia as an approach for cancer therapy, it is now quite feasible to apply hyperthermia to a number of tumour sites and to achieve temperatures that are sufficient to induce a heat shock response. This review will attempt to discuss the current status of hyperthermia-regulated gene therapy, with special emphasis on hyperthermia-regulated immunogene therapy.

New constituents and antiplatelet aggregation and anti-HIV principles of Artemisia capillaris.

Five new constituents including a flavonoid, artemisidin A (1), and four coumarins, artemicapins A (2), B (3), C (4) and D (5), together with 70 known compounds (6-75), have been isolated and characterized from the aerial part of Artemisia capillaris. The structures of these compounds were determined from spectral analyses and/or chemical evidence. Among them, 15 compounds (3, 6, 10, 18. 30-32, 38-41, 44, 45, 51, and 55) showed antiplatelet aggregation activity and three compounds (10, 17, and 51) demonstrated significant activity against HIV replication in H9 lymphocytic cells.

[A study on lyophilization injection of cucurbitacin polylactic acid nanoparticles].

OBJECTIVE: To prepare stable and redispersable Lyophilization injection of cucurbitacin polylactic acid nanoparticles(Cu-PLA-NP). METHOD: An optimal supporting agent was chosen to prepare the Cu-PLA-NP lyophilization injection. The physical and chemical properties of the injection were evaluated. RESULTS: The shape, diameters, association ratio, drug loading and pH of Cu-PLA-NP colloidal solution and lyophilization injection were not changed. The content of water consisted with the requirement of lyophilization injection. The critical relative humidity was 69.52%. The stability was good. CONCLUSION: It is practicable to prepare Cu-PLA-NP lyophilization injection with proper formulas and preparation processes.

[Inhibitory effect of artemin on endotoxin-induced nitric oxide synthesis].

OBJECTIVE: To investigate the effect of Artemin on LPS-induced nitric oxide synthesis in macrophages. METHOD: 1. Nitrite oxide (NO) production of RAW 264.7 cells was induced by LPS or LPS in combination with interferon-gamma (IFN gamma) in the presence or absence of Artemin. The amount of NO in the supernatant of RAW 264.7 cells was detected with Griess reagent. 2. Balb/c mice were injected with Artemin (i.m.) 50 mg.kg-1.d-1 x 3 d, and intraperitoneal macrophages were collected to detect the LPS-induced NO production. RESULT: LPS 1.0, 0.2 microgram.ml-1 or IFN-gamma 100u + LPS 1.0, 0.2, 0.04 microgram.ml-1 could induced a large amount of NO synthesis of RAW 264.7 cells. Artemin showed a significant inhibitory effect on LPS or IFN-gamma + LPS-induced NO production in a dose dependent manner. After treatment with Artemin, the response of Balb/c mice to LPS was reduced, which was showed by a decrease in NO production of intraperitoneal macrophages induced by LPS. CONCLUSION: Artemin could reduce LPS-induced production of inflammatory factors resulting in the inhibition of inflammatory effects.

[Poteintation of vincristine-induced apoptosis by tetrandrine, neferine and dauricine in the human mammary MCF-7 multidrug-resistant cells].

AIM: To investigate the poteintation of vincristine-induecd apoptosis by tetrandrine, neferine and dauricine isolated from Chinese medicinal plants in the human mammary MCF-7 multidrug resistant cells. METHODS: The apoptotic cells were detected by fluorescent staining of a combination of Hoechst 33342 and propidium iodide (PI), flow cytometry and agarose electrophoresis. RESULTS: The apoptotic cells induced by vincristine alone accounted for about 10% of all the cancer cells, while the percentage of apoptotic cells induced by a combination of vincristine with tetrandrine, neferine, or dauricine was found to be significantly higher than that by vincristine alone, and their reversal effects were positively correlated with the drug concentration and the exposure time. In addition, tetrandrine was shown to be the most potent in the reversal efficacy among the three compounds to be tested for apoptosis in vitro. CONCLUSION: Tetrandrine, neferine and dauricine showed obvious potenitiation of vincristine-induced apoptosis in the human mammary MCF-7 multidrug-resistant cells.

Enhancement of radiotherapy by hyperthermia-regulated gene therapy.

PURPOSE: Interleukin 12 (IL-12) has shown strong antitumoral effects in numerous pre-clinical studies and appears to act synergistically with radiation in murine tumors. The major impediment to its clinical use has been its systemic toxicity. While using intratumorally injected viral gene therapy vectors encoding IL-12 reduces systemic side effects substantially, elevated systemic transgene levels are still observed because adenovirus can reach the circulation. Further restricting IL-12 expression in the tumor is therefore desirable in a combined radiation and adenovirus mediated cancer gene therapy regimen. METHODS AND MATERILAS: Hyperthermia-regulated gene therapy was tested in a nonimmunogenic B16.F10 melanoma line that is syngeneic with C57BL/6 mice. For hyperthermic gene therapy, an adenoviral vector coding for IL-12 under the control of the promoter of the human heat shock protein 70B (hsp70B) was used. One week after transplantation (at a 5-7 mm diameter), tumors were irradiated with 3 x 11 Gy (mo-we-fri). Adenovirus was injected at 3 x 10(8) pfu/tumor 24 h before the last radiation fraction or 3 days afterwards. Hyperthermia was performed 24 h later at 42.5 degrees C. Growth delay to reaching 3 times initial tumor volume was chosen as the biologic endpoint. IL-12 levels in tumor and serum were determined by using the enzyme-linked immunosorbant assay (ELISA). RESULTS: Adenovirus mediated intratumoral expression of IL-12 under the control of a heat inducible promoter in combination with hyperthermia is almost as effective as that under the control of a constitutive cytomegaly virus (CMV) promoter while systemic transgene levels are substantially reduced with the heat inducible promoter. The response to radiotherapy is improved considerably when combined with heat inducible gene therapy without apparent systemic toxicity. When used as a single dose, applying IL-12 gene therapy after completion of radiotherapy appears to be beneficial. CONCLUSIONS: Hyperthermia-regulated gene therapy in combination with radiation is feasible and therapeutically effective in murine tumors with no apparent systemic toxicity.

Characterization of risk factors for vascular dementia: the Honolulu-Asia Aging Study.

BACKGROUND: The Honolulu Heart Program (HHP) is a prospective study of heart disease and stroke that has accumulated risk factor data on a cohort of 8,006 Japanese American men since the study began in 1965. A recent examination of the cohort identified all patients with vascular dementia (VaD) using the criteria of the California Alzheimer's Disease Diagnostic and Treatment Center. OBJECTIVE: To characterize patients with VaD by stroke subtype and to investigate risk factors for VaD in a cohort of Japanese American men, aged 71 to 93, living in Hawaii and participating in the HHP. METHODS: Sixty-eight men with VaD were compared with 3,335 men without dementia or stroke (NSND). Men with VaD were also compared with 106 men with stroke who were not demented (SND). Candidate risk factors were measured prospectively. RESULTS: Of the 68 men with VaD there were 34 (50%) whose VaD was attributed to small vessel infarcts, 16 (23%) whose VaD was related to large vessel infarcts, and 11 (16%) with both large and small vessel infarcts. The remainder could not be classified. In a multivariate logistic regression model for VaD compared with NSND containing variables found to be associated with VaD in a univariate analysis, age (odds ratio [OR] 1.19, 95% confidence interval [CI] 1.13 to 1.27), coronary heart disease (OR 2.50, 95% CI 1.35 to 4.62), and 1-hour postprandial glucose (OR 1.41, 95% CI 1.06 to 1.88) remained significantly predictive of VaD, whereas preference for a Western diet (OR 0.54, 95% CI 0.30 to 0.98) as opposed to an Oriental or mixed diet and use of supplementary vitamin E (OR 0.32, 95% CI 0.12 to 0.82) were protective. A similar model for the comparison of men with VaD and SND revealed age (OR 1.24, 95% CI 1.14 to 1.35) was predictive of VaD, whereas preference for a Western diet (OR 0.43, 95% CI 0.22 to 0.86) was protective. CONCLUSIONS: The most common stroke subtype associated with VaD was lacunar stroke. Age and traditional vascular risk factors are important contributors to the development of VaD in late life. The antioxidant vitamin E and presently unknown factors related to a Western diet as opposed to an Oriental diet may be protective against developing VaD.