Clinical features and treatment outcomes of pediatric Langerhans cell histiocytosis with macrophage activation syndrome-hemophagocytic lymphohistiocytosis.

BACKGROUND: Langerhans cell histiocytosis (LCH) is a rare myeloid neoplasm. A few LCH patients had Macrophage activation syndrome-hemophagocytic lymphohistiocytosis (MAS-HLH), a life-threatening, hyper-inflammatory syndrome. We retrospectively described the clinical-biological characteristics of a series of 28 pediatric LCH patients with MAS-HLH in a single center. We further analyzed the difference in treatment outcomes between second-line chemotherapy (cytarabine and cladribine) and targeted therapy (dabrafenib) for BRAF-V600E-positive patients. RESULTS: LCH patients with MAS-HLH were aged < 2 years, harbored high frequencies of risk organ, skin, or lymph nodes involvement, and most of them carried BRAF-V600E mutation in lesions (88.0%) or plasma (90.5%). Patients were firstly treated with the initial induction first-line therapy (vindesine-steroid combination), and most of them (26/28) failed to control the active MAS-HLH after one six-week course of induction treatment. Then they were shifted to second-line chemotherapy or targeted therapy dabrafenib. BRAF-V600E-mutant patients treated with dabrafenib had prompt resolution of MAS-HLH signs and symptoms with less toxicity than second-line chemotherapy. Moreover, the progression-free survival (PFS) rate for patients given dabrafenib was much higher than those treated with chemotherapy (4 year-PFS: 75% vs. 14.6%, P = 0.034). CONCLUSIONS: LCH patients with MAS-HLH harbored specific clinical-biology characteristics compared to the multisystem LCH without MAS-HLH. The BRAF inhibitor dabrafenib provides a promising treatment option for LCH with MAS-HLH.

Significance of serum Th1/Th2 cytokine levels in underlying disease classification of childhood HLH.

OBJECT: Goal of this research was to investigate values of serum cytokines in childhood HLH with different triggers, with the expectation to find secretion spectrum of 5 main types of underlying diseases. METHOD: 118 newly diagnosed HLH were included, and serum concentrations of 6 cytokines were tested before treatment began. Absolute cytokine levels and ratios between them were then studied in the HLH groups collectively and separately RESULTS: In general, IFN-γ, IL-10 and IL-6 showed differences among 5 HLH groups. Specifically, relative levels of these three cytokines to each other were meaningful in distinguishing 4 types of HLH. Level of IL-6 was higher than those of IFN-γ or IL-10 in HLH driven by Systemic auto-inflammatory disorders (SAIDs) or Langerhans Cell Histiocytosis (LCH), while primary HLH and EBV-HLH shared elevated ratio of IL-10 to IL-6. Although more than one distinctive ratios were found in 3 HLH groups, combination of these parameters didn't offer optimal balance between sensitivity and specificity. CONCLUSION: As a group of easily gained laboratory findings, cytokine levels were reliable in the procedure of roughly classifying HLH cases with the help of patients' clinical phenotype. However, adequate data is still needed to explore the significance of these indicators in identifying one particular underlying disease accurately.

The therapeutic effects of Jaceosidin on lipopolysaccharide-induced acute lung injury in mice.

Acute lung injury (ALI) results from various factors including uncontrolled pulmonary inflammation, oxidative damage and the over-activated complement with high mortality rates. Jaceosidin was a flavonoid compound with significant anti-complement activity. We aimed to investigate the therapeutic effects of Jaceosidin on ALI induced by lipopolysaccharide (LPS). Mice were orally administrated with Jaceosidin (15, 30 and 60 mg/kg) after LPS challenge. 24 h after LPS challenge, Jaceosidin could significantly decrease the lung wet-to-dry weight (W/D) ratio and the protein concentration in bronchoalveolar lavage fluid (BALF). Jaceosidin could down-regulate the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1ß (IL-1ß), together with up-regulation the levels of interleukin-4 (IL-4) and interleukin-10 (IL-10) in BALF. Jaceosidin could significantly decrease the levels of myeloperoxidase (MPO), cyclooxygenase-2 (COX-2) and nuclear factor-κB (NF-κB), COX-2 mRNA and NF-κB p65 mRNA together with increasing the activity of catalase (CAT). Additionally, Jaceosidin attenuated lung histopathological changes, inhibited the expressions of COX-2 and NF-κB p65 and reduced complement deposition with decreasing the levels of complement 3 (C3) and complement 3c (C3c) in serum. These data suggest that Jaceocidin may dampen the inflammatory response and decrease the levels of complement together with the antioxidant activity following LPS-induced ALI.

Acute restraint stress triggers progesterone withdrawal and endometrial breakdown and shedding through corticosterone stimulation in mouse menstrual-like model.

Stress impacts the reproductive axis at the level of the hypothalamus and the pituitary gland, which exert an effect on the ovary. Menstruation is regulated by the hypothalamic-pituitary-ovary (HPO) axis. However, the role of stress in menstruation remains unclear. The objective of this study was to explore the role of stress in endometrial breakdown and shedding, using the pseudopregnant mouse menstrual-like model. Female mice were mated with vasectomized males and labeled day 0.5, upon observation of a vaginal seminal plug. On day 3.5, decidualization was induced in pseudopregnant mice using arachis oil. On day 5.5, pseudopregnant mice with artificial decidualization were placed in restraint tubes for 3 h. The findings indicated that acute restraint stress resulted in the disintegration of the endometrium. While corticosterone concentration in the serum increased significantly due to restraint stress, follicle-stimulating hormone (FSH), luteinizing hormone (LH) and progesterone (P4) levels in the serum decreased significantly. An endometrial histology examination indicated that progesterone implants may rescue P4 decline caused by acute stress and block endometrium breakdown and shedding. In addition, mice were treated with metyrapone, an inhibitor of corticosterone synthesis, 1 h prior to being subjected to restraint stress. Interestingly, metyrapone not only inhibited stress-induced endometrium breakdown and shedding, but also prevented stress-induced reduction of P4, LH and FSH. Furthermore, real-time PCR and western blot showed that mRNA and protein expression of CYP11A1 (cytochrome P450, family 11, subfamily A, polypeptide 1) and steroidogenic acute regulatory protein (StAR), the two rate-limiting enzymes for progesterone synthesis in the ovary, decreased following acute stress. But metyrapone prevented the reduction of StAR expression induced by restraint stress. Overall, this study revealed that acute stress results in an increase in corticosterone, which may inhibit LH and FSH release in the serum and CYP11A1 and StAR expression in the ovary, which finally leads to the breakdown and shedding of the endometrium. These experimental findings, based on the mouse model, may enable further understanding of the effects of stress on menstruation regulation and determine the potential factors affecting stress-associated menstrual disorders.

Aberrant TIG1 methylation associated with its decreased expression and clinicopathological significance in hepatocellular carcinoma.

Recently, it has been reported that tazarotene-induced gene 1 (TIG1) methylation was frequently detected in a variety of human cancers. However, the relationship between the TIG1 methylation and the characteristics of hepatocellular carcinoma (HCC) remains unknown. The aim of present study was to observe the promoter methylation of TIG1 in HCC tissues and assess its prognostic significance for HCC. Real-time quantitative polymerase chain reaction and methylation-specific polymerase chain reaction were used, respectively, to examine the mRNA expression and methylation status of TIG1 in 91 pairs of HCC and adjacent noncancerous tissues. The mRNA expression level of TIG1 was significantly lower in HCC tissues than in adjacent noncancerous tissues. The rate of TIG1 promoter methylation was significantly higher in HCC tissues than in adjacent noncancerous tissues (P < 0.001). A strong correlation between downregulation and promoter methylation was found in these tumors (P < 0.001). More importantly, TIG1 methylation status was related to tumor size (P = 0.015), histological differentiation (P = 0.004), and tumor stage (P < 0.001). Kaplan-Meier survival analysis showed that TIG1 promoter hypermethylation was associated with a worse outcome in patients with HCC. Further, Cox multivariate analysis indicated that TIG1 methylation status was an independent prognostic factor for the overall survival rate of HCC patients. In conclusion, our data suggested that epigenetic silencing of TIG1 gene expression by promoter hypermethylation may play an important role in HCC.

[Clinical study of extraction treatment of Class II division I malocclusion with Empower self-ligating brackets].

PURPOSE: To evaluate and compare the treatment efficiency of Empower interactive self-ligating brackets and traditional brackets in Class II division I extraction patients. METHODS: Forty patients with Class II division I malocclusion were randomly divided into 2 groups. Twenty patients received Empower self-ligating technique (group A) and the other 20 patients received MBT technique (group B). Four first premolars were extracted and without any other anchorage devices added in both groups. The duration of treatment, the number of visits and chair-side time were recorded. Cephalometric analysis was performed before and after treatment. The data was analyzed with SPSS 13.0 software package for paired t test. RESULTS: Treatment time and number of visits in group A were more than in group B, but there was no significant difference between the 2 groups. Chair-side time in group A reduced 151.15s on average compared with group B. Significant changes were observed in both groups after treatment. Upper and lower anterior teeth retracted and convex profile improved.U1-SN, U1-NA, L1-MP, L1-NB, UI-PTV, LI-PTV, UL-EP, LL-EP decreased. Significant differences were found in UM-PTV between the 2 groups(P<0.05). CONCLUSIONS: Compared with traditional brackets, Empower self-ligating brackets can save chair-side time, control anterior teeth torque and posterior teeth anchorage effectively, but can not reduce the treatment time or number of visits. Supported by Youth Research Project of Shanghai Municipal Health Bureau(2010Y155).

[The effect of two different methods of rapid maxillary expansion on treatment results of skeletal Class III malocclusion patients with maxillary protraction in early permanent dentition].

PURPOSE: To evaluate and compare the effect of single rapid maxillary expansion and repetitive rapid expansion and constriction with maxillary protraction in treating early permanent skeletal Class III patients. METHODS: Twenty children with skeletal Class III malocclusion were randomly divided into two groups.Ten patients received 1 week of rapid expansion,followed by maxillary protraction(group A) and the other ten patients received 5 weeks of repetitive rapid expansion and constriction followed by maxillary protraction (group B).Cephalometric analysis was performed before and after treatment. The data was analyzed with SPSS 13.0 software package for paired t test. RESULTS: Significant changes in cranio-maxillofacial structures were observed in both groups after 6 months of treatment.Point A moved forward. SNA, ANB, UL-EP, U1-PP, SN-MP, ANS-Me/N-Me, Wit's value increased. L1-MP, LL-EP decreased. Significant differences were found in SNA,ANB,UL-EP between the two groups(P<0.05). CONCLUSIONS: The application of combined repetitive rapid expansion and constriction with maxillary protraction increases the amount of forward movement of the maxilla significantly and presents more favourable profile results in early permanent Class III malocclusion patients.

The nuclear factor-κB pathway is involved in matrix metalloproteinase-9 expression in RU486-induced endometrium breakdown in mice.

BACKGROUND: Progesterone-withdrawal (WP)-induced endometrial breakdown occurs in both physiological and pathological processes such as menstruation and abortion. However, the underlying mechanisms are not clearly understood. As the nuclear factor-κB (NF-κB) pathway has been proposed to play a role in endometrial breakdown, we tested this hypothesis using RU486-induced mouse menstruation-like model. METHODS: The activation of NF-κB was evaluated by immunohistochemistry, western blot and immunofluorescence. The expression of matrix metalloproteinase-9 (MMP9) was analyzed by real-time PCR and its proteins by gelatin zymography and western blot. Chromatin immunoprecipitation was used to investigate the direct binding of NF-κB to MMP9 gene promoter. Inhibitors of NF-κB were used to block its signal in vivo and in vitro to analyze the function of NF-κB in the tissue breakdown process. RESULTS: Administration of RU486 resulted in increased phospho-IκB levels and nuclear translocation of p65 in decidual stromal cells, accompanied by the up-regulation of NF-κB inducing kinase and IκB kinase ß mRNA. The NF-κB inhibitor, 'pyrrolidine dithiocarbamate' partially suppressed the RU486-induced endometrial breakdown, thus verifying the role of this pathway in vivo. MMP9 was up- and down-regulated following the NF-κB activation and inhibition, respectively. RU486 stimulated recruitment of NF-κB p65 to the MMP9 promoter and further increased its expression. Effects of NF-κB activation and inactivation on MMP9 expression were further explored in human stromal cells in vitro. A similar MMP9 expression pattern was observed in cultured human, as well as mouse, decidual stromal cells following RU486 treatment. CONCLUSIONS: The activation of the NF-κB pathway induces downstream target genes, including MMP9 from stromal cells to facilitate tissue breakdown in mouse uterus, highlighting the likelihood that this regulatory pattern exists in the human endometrium.

Identification of a pou2 ortholog in Chinese sturgeon, Acipenser sinensis and its expression patterns in tissues, immature individuals and during embryogenesis.

The class V POU family genes, including pou5f1 and pou2, encode transcription factors critical for the maintenance of pluripotency in embryonic stem cells (ESC) and germ line cells in vertebrates. In the present study, the full-length cDNA of a pou2 ortholog in A. sinensis, Aspou2, was cloned and sequenced. This cDNA sequence is 2,853 base pairs in length and encodes a peptide of 431 amino acid residues. A comparison of the deduced amino acid sequence of Aspou2 with that of other vertebrate species showed that they were highly conserved in the POU domain, which shared 88 and 90% identity with that of zebrafish and medaka, respectively, and was 69, 67 and 67% identical to frog, mouse and human, respectively. RT-PCR analysis revealed that Aspou2 was detected in all tissues examined except for the liver, and high mRNA levels of Aspou2 were found in the muscle, pituitary and brain. During the embryogenesis and early larval development, the expression level of Aspou2 mRNAs decreased gradually apart from 1-day larvae that were not observed. Furthermore, Aspou2 seemed to raise with the development of gonads of immature Chinese sturgeons. These results suggested the possible involvement of Aspou2 in the nonpluripotent cells, pluripotent cells, embryogenesis, and gonad development.

Molecular and expression analysis of apolipoprotein E gene in the Chinese sturgeon, Acipenser sinensis.

Apolipoproteins are carrier proteins that bind to lipids to form lipoprotein particles and have been shown to play an important role in lipid metabolism. In this study, a full-length cDNA for apolipoprotein E, named AsapoE, was cloned from the Chinese sturgeon (Acipenser sinensis). This cDNA sequence is 1289 bp in length, and codes for a polypeptide of 274 amino acid residues, which is 45% and 42% identical to that of the rainbow trout and zebrafish, respectively, and 39%, 30%, and 29% identical to frog, mouse, and human respectively. The predicted AsApoE protein has a conserved amphipathic α-helix region with the potential to bind to lipids. RT-PCR analysis reveals that AsapoE is expressed in all tissues examined with a preferential expression in the kidney and liver. During the embryo development stage, AsapoE mRNA is low but still detectable at gastrula stage embryos; then AsapoE mRNAs reach a higher level in muscle contraction stage embryos, this relatively stable expression persists during the following embryogenic stages and declines 1 day after hatching. These results will serve as a basis for comparative studies on vertebrate apoE genes.

Identification of a vasa homologue gene in grass carp and its expression pattern in tissues and during embryogenesis.

The RNA helicase Vasa is a member of the DEAD box protein family that plays an indispensable role in germ cell determination in eukaryotes such as in Drosophila and Xenopus species. In this study, the grass carp homologue of the Drosophila vasa gene, Civasa (Ctenopharyngodon idella vasa) was obtained using degenerate primers in RT-PCR and 5' and 3' rapid amplification of cDNA end polymerase chain reaction from the grass carp ovary SMART cDNA. This cDNA sequence encodes a 670 amino acid residue protein that contains eight consensus regions for the DEAD box protein family, 9 arginine-glycine repeats and 9 arginine-glycine-glycine repeats, a common character of known Vasa homologues. CiVasa shows high identity to that of zebrafish and other animals, suggesting Vasa is highly conserved through evolution. RT-PCR analysis reveals that in grass carp tissues, both ovaries and testes contain large amounts of vasa gene transcripts whereas no Civasa transcript is detected in somatic tissues examined. The Civasa transcripts are present at a high level from the 2 cell stage to gastrula stages which indicated that Cicasa transcripts are maternally inherited. The predicted protein sequence, localization and conserved pattern of gene expression suggest that Civasa plays an important role in the germ cell determinant and development in grass carp as proposed for other teleost species.

Molecular characterization of Chinese sturgeon gonadotropins and cellular distribution in pituitaries of mature and immature individuals.

Chinese sturgeon (Acipenser sinensis) is a rare and endangered species, and also an important resource for the sturgeon aquaculture industry. To understand molecular characterization of Chinese sturgeon gonadotropins (GTHs), we cloned the full-length cDNAs of gonadotropin subunits common alpha (GTH-alpha), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) from a pituitary cDNA library of mature female. Two subtypes of GTH-alpha were identified. The nucleotide sequences of A. sinensis common alpha I (AsGTH-alpha I), common alpha II (AsGTH-alpha II), FSHbeta (AsFSHbeta) and LHbeta (AsLHbeta) subunit cDNAs are 345, 363, 387 and 414bp in length, and encode mature peptides of 115, 121, 129 and 138aa, respectively. Then, three polyclonal antibodies were prepared from the in vitro expressed AsGTH-alpha I, AsFSHbeta and AsLHbeta mature proteins, respectively. Significant expression differences were revealed between immature and mature sturgeon pituitaries. Western blot detection and immunofluoresence localization revealed the existence of three-gonadotropin subunits (AsGTH-alpha, AsFSHbeta and AsLHbeta) in mature sturgeon pituitaries, but only AsFSHbeta was detected in immature individual pituitaries during early stages in the sturgeon life, and obvious difference was observed between males and females. In males, AsFSHbeta was expressed in 4-year-old individuals, whereas in females, AsFSHbeta was just expressed in 5-year-old individuals.

[Reproductive biology and artificial propagation of Acipenser sinensis below Gezhouba Dam].

A total of 36 females and 21 males of Chinese sturgeon Acipenser sinensis were caught in 1998-2004 excluding 2002 to study the characteristics of their reproductive biology and the effect of their artificial propagation. The results showed that the body length (BL), body mass (BM) and age of the females were 240-320 cm, 140-432 kg, and 15-30 years, and those of the males were 153-284 cm, 70-244 kg and 12-26 years, respectively. The inducing rate was 93.1% for females and 100% for males, and the ova had 7 different colors. The absolute fecundity was 200,000-590,000 eggs, with an average of 358,000 eggs, and the relative fecundity to BM was 820-3,020 eggs per kg, with an average of 1,590 eggs per kg. The sperm had 4 different colors. The absolute sperm quantity obtained from one male was 1,000-5,952 ml, with an average of 2,597.8 ml, and the relative sperm quantity to BM was 1.25-31.24 ml . kg(-1), with an average of 13.3 ml . kg(-1). During the study period, the average fertilization rate in artificial propagation was 63.7%, and the hatching rate was 48.1%, with 4,762,000 fry obtained. Compared with the data in 1976, the natural reproductive capacity of the Chinese sturgeon broodstocks declined greatly.

Rheological and fractal characteristics of granular sludge in an upflow anaerobic reactor.

The rheological and fractal characteristics of the granular sludge in an upflow anaerobic sludge blanket (UASB) reactor were investigated in this study. The influences of sludge concentration and temperature on the rheological properties of the granular sludge were evaluated, and the Bingham model was adopted to describe its rheology. In addition, image analysis was used to determine the sludge fractal dimension. The results indicate that the UASB granular sludge showed a shear-thinning behavior. The relationships between the limiting viscosity and the sludge concentration, as well as the limiting viscosity and temperature could be respectively modeled using an exponential equation and Arrhenius equation well. The Bingham model was able to adequately describe the rheology of the granular sludge. The fractal dimension of the granular sludge, 2.79+/-0.03, was larger than that of some other aggregates, suggesting that the granular sludge were more compact and denser. Furthermore, the relationship between rheological and fractal properties of the granular sludge could be properly described with the model proposed by Shih et al. [1990. Scaling behavior of the elastic properties of colloidal. Phys. Rev. A 42, 4772-4779].