Application of Demirjian's and Cameriere's Method in Dental Age Estimation of 8-16 Year Old Adolescents from Hunan Han Nationality.

ABSTRACT: Objective To apply Demirjian's and Cameriere's method for dental age estimation of adolescents from Hunan Han nationality, and compare the accuracy of the two methods. Methods A total of 480 orthopantomograms of?8-16 year?old adolescents from Hunan Han nationality?with no special diseases and good nutritional status were collected?by Xiangya Stomatological Hospital of Central South University from January, 2016 to July, 2017, among them 236 males and 244 females. The dental age of each adolescent was determined by Demirjian's method and Cameriere's method, respectively, and the paired t-test of the estimated dental age and the chronological age determined by the two methods was conducted by SPSS 20.0 software to compare the difference between estimated dental age and chronological age. Results Mean chronological age of males and females was 11.91 and 11.88 years, respectively. The estimated dental age determined by Demirjian's method showed an underestimate of chronological age by an average of 0.11 years （males） and 0.15 years （females）, while the estimated dental age determined by Cameriere's method showed an underestimate of chronological age by an average of 0.83 years （males） and 0.72 years （females）. Conclusion Demirjian's method is more accurate than Cameriere's method in dental age estimation of adolescents from Hunan Han nationality, therefore more suitable for dental age estimation of adolescents in this region.

[Clinical characteristics and risk factors in pregnancy with severe community-acquired pneumonia].

Objective: To analyze clinical characteristics of severe community-acquired pneumonia during pregnancy and its outcomes, and to explore the relevant risk factors. Methods: From September 2012 to September 2017, 324 398 pregnancies admitted in 7 tertiary hospitals were included. Clinical data of 33 cases of pregnancies with severe community-acquired pneumonia (severe pneumonia group) and 214 cases of pregnancies with common community-acquired pneumonia (control group) were reviewed retrospectively, including the clinical information, manifestations, laboratory examinations and pregnancy outcomes. Relevant risk factors were analyzed by multivariate logistic regression analysis. Results: (1) General data: pregnancies with severe community-acquired pneumonia accounted for 0.010% (33/324 398) of hospitalized pregnancies, the gestational age of two groups were (28±8) and (23±8) weeks, body mass index were (21.7±2.1) and (25.5±3.4) kg/m(2), rate of low income were 54.5% (18/33) and 31.8% (68/214) , respectively. The differences between two groups were all statistically significant (all P<0.05). No significant differences were found in age, pregnancy and parity times, rate of main pregnant complications such as diabetes and hypertension, educational level, asthma and onset seasons between two groups (all P>0.05). (2) Clinical data: the severe pneumonia group had significantly higher incidence of fever [100.0% (33/33) vs 75.2% (161/214) ], shortness of breath (90.9% vs 16.8%) compared with the control group (all P<0.05) .The median peripheral leukocytes counts were 12.3×10(9)/L and 10.2×10(9)/L, the hemoglobin level were (84±18) and (107±14) g/L,the albumin level were (26±4) and (37±3) g/L, the median serum urea nitrogen level were 3.7 and 2.4 mmol/L, the serum creatinine level were (72±25) and (45±11) µmol/L, respectively in two groups. The differences were all statistically significant (all P<0.05). No significantly statistical differences were found in coagulation indicator and cardiac function between two groups (all P>0.05). (3) Treatments: in severe pneumonia group, 12 patients (36.4%,12/33) needed invasive mechanical ventilation, 9 patients (27.3%,9/33) needed non-invasive mechanical ventilation, average time of mechanical ventilation was (7±4) days;8 patients (24.2%,8/33) with septic shock needed vasoactive drugs. However, there was no patient in control group needing mechanical ventilation and vasoactive drugs. (4) Pregnant outcomes: one patient (3.0%,1/33) died in the severe pneumonia group, while no death occurred in the control group. The hospital stay between two groups were (15.1±4.1) and (7.0±1.9) days, the rates of abortion and stillbirth between two groups were 42.4% (14/33) and 3.3% (7/214) , the rates of premature were 10/19 and 6.3% (13/207) , the rates of cesarean were 15/19 and 43.0% (89/207) , the rates of low birth weight newborn were 17/19 and 14.0% (29/207) , the rates of infected newborn were 15/19 and 10.1% (21/207) , the birth weights were (2 165±681) and (3 102±400) g, respectively. The differences between two groups were all statistically significant (all P<0.05). (5) Multivariate logistic regression analysis demonstrated that anemia, low body mass index, hypoproteinemia were risk factors for severe pneumonia in pregnancy (all P<0.05) . Conclusions: Pregnancy with severe community-acquired pneumonia may be complicated by multiple organ dysfunctions, lead to adverse outcomes. Anemia, malnutrition are risk factors for pregnancy with severe pneumonia. Active and effective treatment may improve its prognosis.

Diagnostic and prognostic implications of serum miR-101 in osteosarcoma.

Blood-circulating microRNAs (miRNAs) have been reported to be used as potential biomarkers in various cancers. MiR-101 has been found to act as a tumor suppressor in many tumor types, but little is known for osteosarcoma. The purpose of this study was to investigate miR-101 expression in osteosarcoma patients and assess its correlation with clinical features and prognosis. Serum samples from 152 osteosarcoma patients and 70 healthy controls were detected using quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The data showed that miR-101 expression levels were remarkably underexpressed in serum samples from osteosarcoma patients compared to controls, and the post-treatment serum miR-101 expression was significantly higher than that in the pre-treatment expression. Low serum miR-101 expression was positively associated with advanced clinical stage and distant metastasis. Receiver operating characteristic (ROC) curve analysis showed that serum miR-101 could serve as a useful marker for osteosarcoma diagnosis, with a high sensitivity and specificity. Moreover, patients with high miR-101 expression had longer overall survival and recurrence free survival than those with low miR-101 expression. In addition, both univariate and multivariate analyses showed that serum miR-101 downregulation was associated with shorter overall survival and recurrence free survival. Our present results implicated serum miR-101 might be a useful biomarker for the clinical diagnosis and prognosis of osteosarcoma.

Exposure Duration Is a Determinant of the Effect of Sinusoidal Electromagnetic Fields on Peak Bone Mass of Young Rats.

We proposed a three-step strategy to obtain the optimal therapeutic parameters, which is composed of large-scale screening at cellular level, verification in animal experiments, and confirmation by a clinical trial. The objective of the current study was to test the feasibility of our strategy. Newborn rat calvarial osteoblasts were treated by 50 Hz 1.8 mT sinusoidal electromagnetic fields (SEMFs) with 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 h/days, respectively. The osteogenic differentiation and maturation of the osteoblast were assayed and compared to obtain the optimal duration. One-month-old growing rats were then treated by the same SEMFs with 0.5, 1.5, and 2.5 h/days, respectively, and the peak bone mass was analyzed after 2 months. It was found that the optimal exposure duration to promote the osteogenic differentiation and maturation of osteoblasts was 1.5 h/days, judging by the increasing degrees of ALP activity, calcified nodules formed, the gene and protein expression levels of Runx-2, BMP-2, and Col-I, as well as the expression levels of signaling proteins of the BMP-2/Smad1/5/8 pathway. The highest increase of peak bone mass after 2 months was also obtained by 1.5 h/days, judging by the results of X-ray dual-energy absorptiometry, mechanical property analysis, micro-CT scanning, and serum bone turnover marker examinations. The above results indicated that exposure duration is a determinant for the therapeutic effect of EMFs, and the optimal therapeutic effects only can be obtained by the optimal exposure duration.

Reproductive Biology of Leptocybe invasa Fisher & La Salle (Hymenoptera: Eulophidae).

Leptocybe invasa Fisher & La Salle (Hymenoptera: Eulophidae) is an invasive pest in Eucalyptus plantations around the world. The successful colonization of L. invasa is possibly related to its reproductive biology. The objective of this study was to examine the reproductive biology of L. invasa. In Guangxi Province, the sex ratio (proportion of female, 0.99) of L. invasa was female-dominant throughout the year based on natural and artificial infestation. This result was similar to the ratios observed for other geographic populations in China, including those in Fujian (0.99), Guangdong (0.98), Hainan (0.95), Jiangxi (0.96), and Sichuan (0.99). The offspring sex ratio favored females. A large number of females emerged from the galls produced by females, with few males found. Galls on the petioles and midribs of Eucalyptus plants could be caused by newly emerged females with mature eggs. The lengths of the ovariole, spermatheca, common oviduct, and reproductive glands did not differ among L. invasa females, but their lateral oviducts showed differences from 0 to 42 h after emergence, indicating that this insect is proovigenic. These results could explain why L. invasa populations can rapidly increase in invaded areas.

[The relationship between the recurrence of benign paroxysmal positional vertigo and the level of bone mineral as well as estrogen in postmenopausal women].

Objective: To investigate the relationship between the recurrence of benign paroxysmal positional vertigo(BPPV) and the levels of bone mineral density(BMD) and estrogen in postmenopausal women. Methods: A total of 38 postmenopausal women with recurrent BPPV were recruited as study group, in the First Affiliated Hospital of Soochow University from December 2013 to June 2017. Meanwhile, 49 normal menopausal women were included as control. All patients were natural menopausal for over one year.The patients were diagnosed as BPPV based on results of Dix-Hallpike test and Roll-test, with at least two episodes of recurrent onset. In the subjects, BMD was measured by dual X-ray absorptiometry of lumbar vertebrae. Estrogen levels were obtained by testing serum estradiol (E2) levels in early morning fasting venous blood. In the present study, we compared the level of E2 and the value of BMD in two groups by SPSS 21.0. In the study group, patients with decreased BMD were divided into two groups: treatment and untreated group. The recurrence rate of BPPV was compared between the two groups within 12 months. Results: ①The averagel levels of E2 and BMD in the study group were (16.21±11.00)ng/L and -1.68±0.98) respectively, which were significantly lower than those in the control group (t value was 7.03 and 8.05 respectively, both P<0.05). The averagel levels of E2 and BMD incontrol group were(28.52±6.34)ng/L and -0.18±0.77 respectively. ②The number of patients with decreased BMD in the study group (30 cases) was more than that in control group (6 cases), and the difference was statistically significant (P<0.05). ③ The recurrence rate of BPPV in treatment group [17.6%(3/17)] was significantly lower than that of untreated group [61.5%(8/13)], and the difference was statistically significant (P<0.05). Conclusion: Recurrent BPPV in postmenopausal women usually accompany with low levels of estrogen and BMD. Active treatment is helpful for their recurrence of BPPV.

The gall wasp Leptocybe invasa (Hymenoptera: Eulophidae) stimulates different chemical and phytohormone responses in two Eucalyptus varieties that vary in susceptibility to galling.

Gall-inducing insects produce various types of galls on plants, but little is known about the gall-induction mechanism of these galling insects. The gall wasp Leptocybe invasa Fisher & LaSalle (Hymenoptera: Eulophidae) forms galls of different sizes on several Eucalyptus species. To clarify the physiological responses of Eucalyptus to L. invasa infestation, we measured the dynamics of nitrogen (N), carbon (C), total phenolics, total tannins and four types of phytohormones (zeatin [Z] + zeatin riboside [ZR], gibberellins [GA], indole-3-acetic acid [IAA] and abscisic acid [ABA]) in galled and ungalled leaf tissues of two Eucalyptus horticultural varieties (DH201-2 [Eucalyptus grandis × Eucalyptus camaldulensis] and EA [Eucalyptus exserta]) with different susceptibility to galling throughout the larval developmental stages. Nitrogen, total phenolics, tannins and four kinds of phytohormones strongly accumulated in tissues galled by L. invasa (especially during early larval feeding stages). While N, Z + ZR and GA levels were higher, tannins and ABA levels were lower in the galled tissues on the highly susceptible variety. Nitrogen, total phenolics, GA, Z + ZR and IAA levels in the galled tissues gradually decreased during gall development, but ABA and tannins conversely increased in the galled tissues of the less susceptible variety. Our results suggest that the effects of gall-inducing insects on plants depend not only on the susceptibility of the plant infested but also on the developmental stage of galled tissues. Gall formation process is thus synergistically influenced by both gall-inducing insect and plant genotypes.

Expression and significance of miR-21 in multiple myeloma patients.

The aim of the present study is to examine the expression level of peripheral mir-21 in multiple myeloma (MM) patients and to determine its clinical significance. MM patients (30), monoclonal gammopathy of undetermined significance (MGUS) patients (14), and normal controls (20) were recruited to determine the serum level of ß2-MG, IgA and IgM, IgG, λ, κ, TP, ALB, Hb, LDH, and Ca(2+). Gene expression of mir-21 was quantified by SYBR green real-time fluorescent quantitative PCR. We found that the expression level of serum mir-21 in the MM group was significantly higher than the MGUS group and the NC group (P < 0.01). According to the ISS installment, the level of mir-21, lgG, κ, and ALB in the MM group in stage I differed from that in stages II and III. The level of IgA, ß2-MG in stage III was higher as compared with stage I and II (P < 0.05 and P < 0.01).The levels of mir-21, κ, (κ+λ), IgG, (IgG + IgA + IgM), and ß2-MG in MM patients were positively correlated with ALB (P < 0.01). Based on the results, miR-21 plays an important role as an oncogene. Mir-21 may be important in the occurrence, development, and disease prognosis of MM.

[The role of video nystagmography in the diagnosis and treatment of multiple benign paroxysmal positional vertigo].

Objective:To explore the role of video nystagmography in the diagnosis and treatment of multiple benign paroxysmal positional vertigo. Method: Eleven patients of 313 patients with benign paroxysmal positional vertigo had been diagnosed as multiple benign paroxysmal positional vertigo.They were post and horizontal semicircular canal ipsilaterally, bilateral post semicircular canal, superior and horizontal semicircular canal ipsilaterally, and bilateral superior semicircular canal benign paroxysmal positional vertigo.Result:The patients were performed Dix-Hallpike test and Roll test under VNG, combined with track record by video nystagmography to confirm the affected sites. Six cases were post and horizontal semicircular canal benign paroxysmal positional vertigo ipsilaterally. Three cases were bilateral posterior semicircular canal benign paroxysmal positional vertigo, one case of superior semicircular canal and horizontal semicircular canal benign paroxysmal positional vertigo ipsilaterally, and one case of bilateral superior semicircular canal benign paroxysmal positional vertigo. The symptom of patients got relief after repositioning sequentially. We repositioned the affected canal with strong nystagmus and vertigo at the first time. Conclusion:Multiple benign paroxysmal positional vertigo was rare and easily misdiagnosed because of complex nystagmus. We can confirm the affected canal and intensity by video nystagmography and get good prognosis after repositioning sequentially.

Epidemiological and clinical characteristics of severe fever with thrombocytopenia syndrome (SFTS) in China: an integrated data analysis.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease that was caused by a novel bunyavirus, SFTSV. The study aimed to disclose the epidemiological and clinical characteristics of SFTSV infection in China so far. An integrated clinical database comprising 1920 SFTS patients was constructed by combining first-hand clinical information collected from SFTS sentinel hospitals (n = 1159) and extracted data (n = 761) from published literature. The considered variables comprised clinical manifestations, routine laboratory tests of acute infection, hospitalization duration and disease outcome. SFTSV-IgG data from 19 119 healthy subjects were extracted from the published papers. The key clinical variables, case-fatality rate (CFR) and seroprevalence were estimated by meta-analysis. The most commonly seen clinical manifestations of SFTSV infection were fever, anorexia, myalgia, chill and lymphadenopathy. The major laboratory findings were elevated lactate dehydrogenase, aminotransferase, followed by thrombocytopenia, lymphocytopenia, elevated alanine transaminase and creatine kinase. A CFR of 12·2% was estimated, significantly higher than that obtained from national reporting data, but showing no geographical difference. In our paper, the mortality rate was about 1·9 parts per million. Older age and longer delay to hospitalization were significantly associated with fatal outcome. A pooled seroprevalence of 3·0% was obtained, which increased with age, while comparable for gender. This study represents a clinical characterization on the largest group of SFTS patients up to now. A higher than expected CFR was obtained. A wider spectrum of clinical index was suggested to be used to identify SFTSV infection, while the useful predictor for fatal outcome was found to be restricted.

Discrimination of novel bunyavirus infection using routine laboratory test.

The wide epidemic and high case fatality rate have made severe fever with thrombocytopenia syndrome (SFTS) a significant public health problem. The diagnosis and discrimination of SFTS virus (SFTSV) infection at an early stage of the disease is important for treatment choice. A prospective study was performed in an SFTS reference hospital during 2011-2013. Suspected SFTS patients were recruited and prospectively observed. Comparison between SFTSV-positive and -negative patients was made to identify the parameters that were related to positive detection by discriminant and classification tree analysis. A total of 538 SFTSV-positive and 396 negative patients were recruited and observed. Multiple logistic regression models demonstrated the significant parameters associated with positive detection, including decreased platelet counts and elevated aspartate aminotransferase (AST) level during the first stage (1∼4 days), decreased white blood cell and platelet counts, elevated creatine kinase (CK) and AST levels during the second stage (5∼7 days), and older age, decreased consciousness and elevated CK and AST during the third stage (8-11 days). The classification trees disclosed that the significant predictors for positive SFTSV detection were AST >50.6 U/L and AST/alanine transaminase (ALT) >1.3 at the first stage, CK >257 U/L or 57.7 U/L < CK ≤98.5 U/L with AST/ALT >1.6 at the second stage, as well as CK >630.7 U/L or 114.3 U/L < CK ≤630.7 U/L with decreased consciousness at the third stage. In making the clinically probable diagnosis of SFTS, the supplementation of AST and CK evaluations might remarkably improve the diagnostic capacity of routine laboratory tests, while the leukopenia is of limited use.

Change of selenium in environment and risk of adolescent idiopathic scoliosis: a retrospective cohort study.

OBJECTIVES: Scoliosis is the disease which has a long history over one century. However, the pathogenesis remains unclear at present. To demonstrate the effect of different selenium content in environment on the morbidity of adolescent idiopathic scoliosis (AIS). METHODS: Retrospective cohort study (follow-up from 1997 to 2009): compare the difference morbidity between high selenium group and the normal selenium group of AIS. PATIENTS: 9998 cases from three areas in China were participated in this study. There is different selenium content in these three areas. RESULTS: High selenium levels were significant associated with the AIS morbidity. While low selenium level had no significant correlation with the AIS morbidity. CONCLUSIONS: This study confirmed that high selenium content in the environment was one of risk factors for idiopathic scoliosis. We speculated that the excessive growth of the spine and the spinal cord asynchronous growth effect were key factors that high selenium content in the environment leads to scoliosis.

Effects of raloxifene on cognition, mental health, sleep and sexual function in menopausal women: a systematic review of randomized controlled trials.

BACKGROUND AND OBJECTIVE: Raloxifene has been used as therapy for osteoporosis and ER(+) breast cancer prevention in menopausal women. However, its effects on cognition and climacteric syndrome are controversial. This study reviews the relevant studies and reaches a comprehensive conclusion. METHODS: We retrieved thirteen electronic databases, read the titles and abstracts to exclude ineligible articles, and then read the full text and references to form a basis for decisions using the inclusion criteria. If full text was not available, we asked the author for a copy of the article. After the data were extracted and recorded, the research quality was evaluated by two authors using the Jadad score and Cochrane handbook. RESULTS: We found seven eligible studies. The design, evaluated items, questionnaires and scales were heterogeneous. The design quality was fair as evaluated by the Cochrane Handbook. We found that 60 mg/day raloxifene could improve verbal memory, and 120 mg/day raloxifene produced a 33% decrease in the risk of mild cognitive impairment and also lowered the risk of Alzheimer's disease. There was not enough evidence to state if raloxifene had any effect on depression, anxiety, sleep, sexual function, vasomotor symptoms, but significantly worsened menstrual symptoms. CONCLUSION: Raloxifene may have some benefit for cognition, but it is not significant effect on anxiety, depression, sleep, sexual function, vasomotor symptoms and worsens menstrual symptoms. This drug is safe for treating osteoporosis and preventing breast cancer in menopausal women, but it is not suitable for patients who have any arterial stenosis or thrombophilia.

The microRNA-29 plays a central role in osteosarcoma pathogenesis and progression.

Osteosarcoma is the most common type of bone cancer, with a peak incidence in the early childhood. The relationship between microRNAs (miRNAs) and cancer development attracted more and more attention over the last few years. Members of the miRNA-29 family, including miRNA-29a, miRNA-29b, and miRNA-29c were shown to participate in the development of rhabdomyosarcoma and hepatocarcinogenesis. Here, it has been demonstrated miRNA-29a and miRNA-29b expression levels to be downregulated in most of the osteosarcoma tissues (23 from 30). Besides, miRNA-29a displayed ability to induce apoptosis in both U2OS and SAOS-2 osteoblastic cells. While miRNA-29 members induced apoptosis through p53 gene activation, the effect of miRNA-29a on osteoblastic cells was independent on p53 expression level. Moreover, Bcl-2 and Mcl-1 were earlier demonstrated to be the direct targets of miRNA-29 in many types of cancer tissues and cancers. In both U2OS and SAOS-2 osteoblastic cell types, overexpression of miRNA-29a also downregulated Bcl-2 and Mcl-1, while silencing of miRNA-29a increased their expression. In addition, enhanced expression of miRNA-29a increased the expression of two tumor suppressor genes, E2F1 and E2F3. In summary, data obtained highlight the role of miRNA-29a in the regulation of osteoblastic cell apoptosis by silencing Bcl-2 and Mcl-1 and inducing E2F1 and E2F3 expression.

Protection abilities of influenza B virus DNA vaccines expressing hemagglutinin, neuraminidase, or both in mice.

Every year, a vaccination against Influenza B virus (IBV) is essential due to an antigenic variation. Development of an efficient and convenient vaccine is important for the prevention of viral infection. This study reports examination of the protective immunity in mice evoked by a single inoculation of plasmid DNA expressing hemagglutinin (HA DNA) or neuraminidase (NA DNA) of IBV. The HA DNA or NA DNA was injected intramuscularly into BALB/c mice separately or as a mixture. The injection of plasmid was followed by an electroporation close to the site of puncture. Four weeks later, the immunized mice were challenged with a lethal dose of IBV. The protective abilities of DNA vaccines were evaluated by the detection of specific antibodies in serum, survival rate, virus titer in lungs, and change of body weight. We found that a single dose of HA DNA or NA DNA induced the formation of specific antibodies and conferred effective protection against the lethal challenge of IBV. However, the combined vaccine HA DNA and NA DNA enhanced the protective ability of immunized mice. The obtained results suggested that immunization with single dose of HA DNA, NA DNA or with combination of both could be an efficient method for preventing IBV infection.

Activation of 12-lipoxygenase in proinflammatory cytokine-mediated beta cell toxicity.

AIMS/HYPOTHESIS: Beta cell inflammation and cytokine-induced toxicity are central to autoimmune diabetes development. Lipid mediators generated upon lipoxygenase (LO) activation can participate in inflammatory pathways. 12LO-deficient mice are resistant to streptozotocin-induced diabetes. This study sought to characterise the cellular processes involving 12LO-activation lipid inflammatory mediator production in cytokine-treated pancreatic beta cells. METHODS: Islets and beta cell lines were treated with a combination of IL-1beta, IFN-gamma and TNF-alpha, or the 12LO product 12(S)-hydroxyeicosatetraenoic acid (HETE). Insulin secretion was measured using an enzyme immunoassay, and cell viability was evaluated using an in situ terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling assay. 12LO activity was evaluated and 12LO protein levels were determined using immunoblotting with a selective leucocyte type 12LO antibody. Cellular localisation of 12LO was evaluated using immunocytochemistry. RESULTS: Basal expression of leucocyte type 12LO protein was found in human and mouse islets and in several rodent beta cell lines. In mouse beta-TC3 cells, and in human islets, cytokines induced release of 12-HETE within 30 min. Cytokine addition also induced a rapid translocation of 12LO protein from the cytosol to the nucleus of beta-TC3 cells as shown by subcellular fractionation and immunostaining. Cytokine-induced cell death and inhibition of insulin secretion were partially reversed by baicalein, a 12LO inhibitor. 12(S)-HETE inhibited beta-TC3 cell insulin release in a time- and concentration-dependent manner. Incubating beta-TC3 cells with 100 nmol/l of 12(S)-HETE resulted in a 57% reduction in basal insulin release (6 h), and a 17% increase in cell death (18 h) as compared with untreated cells. 12(S)-HETE activated the stress-activated protein kinase c-Jun N-terminal kinase and p38 within 15 min, as judged by increased kinase protein phosphorylation. CONCLUSIONS/INTERPRETATION: The data suggest that inflammatory cytokines rapidly activate 12LO and show for the first time that cytokines induce 12LO translocation. The effects of 12-HETE on insulin secretion, cytotoxicity and kinase activation were similar to the effects seen with cytokines. The results provide mechanistic information of cytokine-induced toxic effects on pancreatic beta cells and support the hypothesis that blocking 12LO activation could provide a new therapeutic way to protect pancreatic beta cells from autoimmune injury.

The anti-inflammatory compound lisofylline prevents Type I diabetes in non-obese diabetic mice.

AIMS/HYPOTHESIS: Pro-inflammatory cytokines are increased during the active stages of Type I (insulin-dependent) diabetes mellitus. The aim of this study was to investigate the applicability of using a new anti-inflammatory compound, Lisofylline, to prevent diabetes in non-obese diabetic (NOD) mice. Lisofylline has previously been shown to block Th1 cell differentiation and to reduce IL-1 beta-induced dysfunction in rat islets. METHODS: Lisofylline was added to isolated NOD islets in vitro, with or without IL-1 beta. Insulin secretion and DNA damage of the islets was assessed. Lisofylline was administered to female non-obese diabetic mice starting at 4, 7 and 17 weeks of age for 3 weeks. Cytokines and blood glucose concentrations were monitored. Histology and immunohistochemistry were carried out in pancreatic sections. Splenocytes isolated from donor mice were intravenously injected into immunodeficient NOD (NOD.scid) mice. RESULTS: In vitro, Lisofylline preserved beta-cell insulin secretion and inhibited DNA damage of islets in the presence of IL-1 beta. In vivo, Lisofylline suppressed IFN-gamma production, reduced the onset of insulitis and diabetes, and inhibited diabetes after transfer of splenocytes from Lisofylline-treated donors to NOD.scid recipients. However, cotransfer of splenocytes from both Lisofylline-treated and diabetic NOD donors did not suppress diabetes in recipient mice. CONCLUSION/INTERPRETATION: Lisofylline prevents the onset of autoimmune diabetes in NOD mice by a mechanism that does not seem to enhance the function of regulatory T cells, but could be associated with suppression of proinflammatory cytokines and reduction of cellular infiltration in islets. This study suggests that Lisofylline could have therapeutic benefits in preventing the onset of Type I diabetes.

Prevention of autoimmune recurrence and rejection by adenovirus-mediated CTLA4Ig gene transfer to the pancreatic graft in BB rat.

Type 1 diabetes is the result of a selective destruction of pancreatic islets by autoreactive T-cells. Therefore, in the context of islet or pancreas transplantation, newly transplanted beta-cells are threatened by both recurrent autoimmune and alloimmune responses in recipients with type 1 diabetes. In the present study, using spontaneously diabetic BB rats, we demonstrate that whereas isolated islets are susceptible to autoimmune recurrence and rejection, pancreaticoduodenal grafts are resistant to these biological processes. This resistance is mediated by lymphohematopoietic cells transplanted with the graft, since inactivation of these passenger cells by irradiation uniformly rendered the pancreaticoduodenal grafts susceptible to recurrent autoimmunity. We further studied the impact of local immunomodulation on autoimmune recurrence and rejection by ex vivo adenovirus-mediated CTLA4Ig gene transfer to pancreaticoduodenal grafts. Syngeneic DR-BB pancreaticoduodenal grafts transduced with AdmCTLA4Ig were rescued from recurrent autoimmunity. In fully histoincompatible LEW-->BB transplants, in which rejection and recurrence should be able to act synergistically, AdmCTLA4Ig transduced LEW-pancreaticoduodenal allografts enjoyed markedly prolonged survival in diabetic BB recipients. In situ reverse transcription-polymerase chain reaction revealed that transferred CTLA4Ig gene was strongly expressed in both endocrine and exocrine tissues on day 3. These results indicate the potential utility of local CD28-B7 costimulatory blockade for prevention of alloimmune and autoimmune destruction of pancreatic grafts in type 1 diabetic hosts.