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C80H38Er2F16N8O18, triclinic, P 1 ‾ $P\overline{1}$ (no. 2), a = 11.4228(4) Å, b = 12.6712(3) Å, c = 13.2663(4) Å, α = 102.660(2)°, β = 98.047(3)°, γ = 105.509(3)°, V = 1764.54(10) Å3, Z = 1, R gt(F) = 0.0311, wR ref (F 2) = 0.0668, T = 293(2) K.

C40H19EuF8N4O9, triclinic, P 1 ‾ $P\overline{1}$ (no. 2), a = 11.3326(4) Å, b = 12.7652(4) Å, c = 13.2629(4) Å, α = 102.266(2)°, β = 97.689(2)°, γ = 105.630(3)°, V = 1767.93(10) Å3, Z = 2, R gt (F) = 0.0329, wR ref (F 2) = 0.0648, T = 293(2) K.

C80H38Dy2F16N8O18, triclinic, P 1 ‾ $P\overline{1}$ (no. 2), a = 11.3924(5) Å, b = 12.7091(4) Å, c = 13.2583(5) Å, α = 102.496(3)°, β = 97.955(4)°, γ = 105.588(4)°, V = 1765.36(12) Å3, Z = 2, Rgt (F) = 0.0298, wRref (F 2) = 0.0606, T = 293(2) K.

C21H11F4NO4, triclinic, P 1 ‾ $P\overline{1}$ (no. 2), a = 9.5000(4) Å, b = 13.9989(7) Å, c = 14.1423(5) Å, α = 91.355(4)°, β = 108.542(4)°, γ = 98.958(4)°, V = 1756.02(14) Å3, Z = 4, R gt (F) = 0.0494, wR ref (F 2) = 0.1157, T = 293(2) K.

Dasatinib, an orally administered Src-family kinase inhibitor, is combined with the standard chemotherapeutic regimen to enhance antineoplastic activity against core-binding factor acute myeloid leukemia (CBF-AML) in adults; however, limited data are available for use in children. In the present study, we studied the pharmacokinetics and safety of dasatinib in children. Dasatinib (60 or 80 mg/m2 once daily) was administered to 20 children with CBF-AML. Blood samples were collected and drug concentrations were quantified by high-performance liquid chromatography–tandem mass spectrometry (HPLC-MS/MS). Population pharmacokinetic analysis and Monte-Carlo simulations were performed using NONMEM software, and safety analyses were assessed according to the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI-CTCAE) version 4.0 (NCT03844360). Twenty pediatric patients (3.3–14.4 years of age) were included, and a total of 40 dasatinib concentrations were available for population pharmacokinetic analysis. The mean (standard deviation) of the estimated area under the concentration-time curve extrapolated to steady state (AUCss) of dasatinib 60 and 80 mg/m2 was 366.1 (146.6) ng·h/mL and 425.3 (150.7) ng·h/mL, respectively. The majority of adverse events were grade 1/2 in severity, including thrombocytopenia, rash, and pain in the extremities. The estimated cumulative incidence of complete remission and complete molecular response were 95.0% and 75.5%, respectively. The population pharmacokinetics of orally administered dasatinib were evaluated in pediatric CBF-AML patients. The AUCss of dasatinib (80 mg/m2) in CBF-AML pediatric patients was similar to those of dasatinib (100 mg) in adult patients. Dasatinib is well-tolerated in pediatric patients with CBF-AML.

Objective. This research paper is based on a retrospective case-control study for exploring the effects of medical nursing integration and the continuous 4C nursing model to improve the clinical treatment and nursing quality of patients with acute stroke. Method. For this purpose, a total of 313 patients with acute stroke, treated in our hospital from February 2020 to April 2021, were enrolled. They were divided into control and study groups with an even number of patients. The control group received integrated medical care number (N = 156), while the study group received integrated medical care and a continuous 4C nursing model (N = 157). In integrated medical care, the general data, self-nursing ability, degree of neurological impairment, Fugl–Meyer Assessment (FMA) score, Barthel index score, and quality of life score were compared between the two groups. Result. The self-nursing concept, self-nursing responsibility, self-nursing skills, health knowledge, and total score of the patients in the study group were higher than those in the control group ( P P P P

Ovarian cancer (OV) is the most lethal gynecologic malignancy. One major reason of the high mortality of the disease is due to platinum-based chemotherapy resistance. Increasing evidence reveal the important biological functions and clinical significance of zinc finger proteins (ZNFs) in OV. In the present study, the relationship between the zinc finger protein 76 (ZNF76) and clinical outcome and platinum resistance in patients with OV was explored. We further analyzed ZNF76 expression via multiple gene expression databases and identified its functional networks using cBioPortal. RT-qPCR and IHC assay shown that the ZNF76 mRNA and protein expression were significantly lower in OV tumor than that in normal ovary tissues. A strong relationship between ZNF76 expression and platinum resistance was determined in patients with OV. The low expression of ZNF76 was associated with worse survival in OV. Multivariable analysis showed that the low expression of ZNF76 was an independent factor predicting poor outcome in OV. The prognosis value of ZNF76 in pan-cancer was validated from multiple cohorts using the PrognoScan database and GEPIA 2. A gene-clinical nomogram was constructed by multivariate cox regression analysis, combined with clinical characterization and ZNF76 expression in TCGA. Functional network analysis suggested that ZNF76 was involved in several biology progressions which associated with OV. Ten hub genes (CDC5L, DHX16, SNRPC, LSM2, CUL7, PFDN6, VARS, HSD17B8, PPIL1, and RGL2) were identified as positively associated with the expression of ZNF76 in OV. In conclusion, ZNF76 may serve as a promising prognostic-related biomarker and predict the response to platinum in OV patients.

Background Children with acute lymphoblastic leukemia (ALL) undergoing chemotherapy experience a relatively high risk of infection. And the disturbance of gut microbiota is generally believed to impair intestinal barrier function and may induce bacterial infections and inflammation. The study aimed to investigate the alterations in the gut microbiota and assess its relationship with chemotherapy-induced pneumonia in pediatric ALL patients. Methods We conducted a case–control study with 14 cases affected by pneumonia and 44 unaffected subjects and characterized the physiological parameters and gut microbiota by microarray-based technique. Results There were significant differences in α- and β-diversity in the affected group compared with the control group. At species level, the LEfSe analysis revealed that Enterococcus malodoratus, Ochrobactrum anthropi and Actinomyces cardiffensis were significantly abundant in the affected subjects. A receiver operating characteristic (ROC) curve yielded the area under the curve (AUC) of 0.773 for classification between the two groups. In addition, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways involved in the bacterial secretion system were more enriched in the affected group than in the control group. Conclusions Gut microbiota alteration was associated with chemotherapy-induced pneumonia in pediatric ALL patients, which provided a new perspective on the personalized clinical care of pediatric ALL.

Dasatinib, an orally administered Src-family kinase inhibitor, is combined with the standard chemotherapeutic regimen to enhance antineoplastic activity against core-binding factor acute myeloid leukemia (CBF-AML) in adults; however, limited data are available for use in children. In the present study, we studied the pharmacokinetics and safety of dasatinib in children.Dasatinib (60 or 80 mg/mTwenty pediatric patients (3.3-14.4 years of age) were included, and a total of 40 dasatinib concentrations were available for population pharmacokinetic analysis. The mean (standard deviation) of the estimated area under the concentration-time curve extrapolated to steady state (AUCThe population pharmacokinetics of orally administered dasatinib were evaluated in pediatric CBF-AML patients. The AUC

Li Zhang,1,* Xin-Mei Yang,2,* Jing Chen,3 Lei Hu,4 Fan Yang,5 Yue Zhou,5 Bei-Bei Zhao,1 Wei Zhao,2,5,* Xiao-Fan Zhu1,* 1Department of Pediatric, State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, People’s Republic of China; 2Department of Clinical Pharmacy, Clinical Trial Center, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Jinan, People’s Republic of China; 3Pediatric Hematology & Oncology, Shanghai Children’s Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China; 4Tianjin Institute of Pharmaceutical Research (TIPR), Tianjin, People’s Republic of China; 5Department of Clinical Pharmacy, Key Laboratory of Chemical Biology (Ministry of Education), School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xiao-Fan ZhuDepartment of Pediatric, State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, 288 Nanjing Road, Tianjin, 300020, People’s Republic of ChinaTel/Fax +86 22 2390 9001Email xfzhu@ihcams.ac.cnWei ZhaoDepartment of Clinical Pharmacy, School of Pharmaceutical Sciences, Shandong University, No. 44, Wenhua West Road, Jinan, Shandong Province, People’s Republic of ChinaTel/Fax +86 531 8838 3308Email zhao4wei2@hotmail.comBackground: An oral tetra-arsenic tetra-sulfide (AS4S4) formula has been recommended as an outpatient post-remission treatment for Chinese adults with acute promyelocytic leukemia (APL) but limited data are available for children. In this exploratory study, we aimed to evaluate the pharmacokinetics and safety of the AS4S4 formula in children.Methods: Eleven newly diagnosed and one relapsed pediatric patient (4– 14 years of age) treated with the AS4S4 formula were included. Blood samples were collected from 12 children, and drug concentrations were quantified by ICP-MS. Population pharmacokinetic analysis and Monte-Carlo simulation were performed using NONMEM software. Toxic effects were graded according to the NCI-CTCAE, Version 3.Results: A total of 107 arsenic concentrations (0.1– 75.0 μg L− 1) were used for population pharmacokinetic analysis. The median (range) of estimated weight-normalized CL and volume distribution at steady-state were 45.26 (35.63– 82.18) L h− 1 kg− 1 and 230.37 (85.96– 495.68) L kg− 1, respectively. No patients discontinued AS4S4 treatment owing to adverse events, and there were no drug-related adverse events over grades 3– 4. All newly diagnosed APL patients were in MCR with a median follow-up of 28 months (range, 23 to 37 months). Both the estimated 3-year EFS and OS rates were 100%.Conclusion: The pharmacokinetics and safety oral AS4S4 formula was evaluated for the first time in pediatric APL. The pharmacokinetic assessment demonstrated that the dosing regimen of 60 mg/kg/d TID resulted in a higher steady-state through concentration in children than that which was achieved in adults. The results of this study indicate that the AS4S4 formula is safe in newly diagnosed pediatric APL patients.Keywords: acute promyelocytic leukemia, Realgar-Indigo Naturalis Formula, pediatric, safety, population pharmacokinetics

To study the specific immunotherapeutic effect of Dermatophagoides pteronyssinus group 1 major allergen T-cell fusion epitope peptide vaccine TAT-IhC-DPTCE against allergic asthma.One hundred and twenty SPF-grade BALB/c mice were randomized into PBS group (group A), asthma group （group B）, and immune treatment groups respectively receiving intraperitoneal (i.p.) injections of ProDer p 1 allergen (group C), DPTCE （group D）, TAT-DPTCE (group E) or TAT-IhC-DPTCE (group F) （n=20 in each group）. In detail, PBS （group A） or allergen extract derived from Dermatophagoides pteronyssinus （groups B-F, 10 μg） was intraperitoneally injected on days 0, 7 and 14, and was continued by aerosol inhalation from day 21 for 7 consecutive days （0.5 μg/ml, once/day, 30 min each time）. The mice in groups C-F received i.p. injections of 100 μg/ml ProDer p 1, DPTCE, TAT-DPTCE and TAT-IhC-DPTCE respectively 30 min prior to inhalation challenge on days 25-27 as a specific immunotherapy, while those in groups A and B received 200 μl PBS. Twenty-four hours after the last inhalation challenge, all the mice were sacrificed. The lung histopathological changes were examined by HE staining. The levels of IFN-γ, IL-13, IL-10 and TGF-β in the bronchoalveolar lavage fluid (BALF) was determined with ELISA, and eosinophils in the BALF were counted (n=20 mice in each group). The serum level of IgE, IgG1 and IgG2a in orbital blood was determined by ELISA（n=5 mice in each group）.HE staining revealed increased BALF eosinophils and decreased pulmonary inflammation in group F compared with group B. The IFN-γ level in group F ［（298.75±26.09） pg/ml］ was significantly higher than those in groups B［（158.71±20.89） pg/ml］, C［（210.38±18.92） pg/ml］, D ［（229.44±13.00） pg/ml］ and E［（233.24±20.39） pg/ml］ （all P0.01）. Similar results were also found for IL-10 and TGF-β, while the IL-13 levels in groups C ［（47.35±4.71） pg/ml］, D ［（41.90±4.28） pg/ml］, E［（41.05±6.50） pg/ml］ and F［（18.53±5.67） pg/ml］ were all significantly lower than that in group B ［（66.68±6.63） pg/ml］（all P0.01）. The number of BALF eosinophils in group B ［5.65±0.91］×105/ml］ was significantly higher than that in group A ［（0.45±0.39）×105/ml］ （P0.01）, while the BALF eosinophils in groups C ［（4.00±0.59）×105/ml］, D ［（3.39±0.63）×105/ml］, E ［（3.24±0.69）×105/ml］ and F ［（1.42±0.49）×105/ml］ decreased after immune treatment (all P0.01). ELISA results showed that the serum IgE level in group F ［（5.26±1.72） ng/ml］ was significantly lower than those in group B ［（32.81±2.98） ng/ml］ and the other 3 treatment groups [group C, （20.06±3.17） ng/ml; D, （17.06±3.18） ng/ml; E, （16.23±3.61） ng/ml］. Similar results were also obtained for IgG1. In contrast, the serum IgG2a level in group F［（43.10±1.34） ng/ml］ was significantly higher than those in group B［（12.61±1.87） ng/ml］ and the other 3 treatment groups ［group C, （23.37±2.67） ng/ml; D, （25.60±2.10） ng/ml; E, （25.91±1.33） ng/ml］ （all P0.01）.Immunotherapy with chimeric TAT-IhC-DPTCE can effectively ameliorate the allergic airway response and pulmonary inflammation in mice.

The first two authors contributed equally * The last two authors contributed equally Background: Realgar-Indigo Naturalis Formula (RIF), the only commercially oral arsenic agent, was launched in China from 2009. It has been recommended as consolidation or maintenance therapy in adults with acute promyelocytic leukemia (APL). Recently, RIF had been off-label used in pediatric APL in China. However, the knowledge of efficacy and dosage of RIF in pediatric was limited until now. Objective: The purpose of this pilot study was to evaluate effectiveness and pharmacokinetics of RIF as consolidation or maintenance therapy for newly diagnosed and relapsed APL in pediatric after induction therapy with intravenous arsenic trioxide. Meanwhile, modeling and simulation techniques were used to evaluate and optimize RIF dosing regimen in pediatric population. Patients and Methods: A total of 11 newly diagnosed APL and 1 relapsed patient (4-14years of age) were included from July 2016 to August 2017.The one relapsed and six newly diagnosed patients were treated with RIF (60 mg/kg/day TID) as maintenance therapy. Five newly diagnosed patients were treated with RIF (60 mg/kg/day TID) as consolidation and maintenance therapy. The treatment protocol and MRD results are shown in Figure 1. Event-free survival and overall survival were used to evaluate the efficacy. The plasma concentration of arsenic was quantified by inductively coupled plasma mass spectrometry (ICP-MS). Population pharmacokinetic analysis was carried out using the nonlinear mixed effects modeling program NONMEM V 7.2 (Icon Development Solutions, USA). Results: All newly diagnosed APL patients were alive as of July 1, 2019 and were in MCR with a median follow-up of 28 months (range, 23 to 37 months). Both the estimated 3-year EFS and OS rates were 100%. Of note, all the patients completed the postremission therapy containing RIF on an outpatient basis. A total of 107 arsenic concentrations were used for population pharmacokinetic model building. The arsenic concentrations of the blood samples ranged from 0.1 to 75.0 µg/L. The steady-state trough plasma arsenic concentration during the RIF treatment was 47.43μg/L (range, 25.74 to 62.97μg/L).A one-compartment model with first-order elimination fitted the data. Body weight was the only significant covariate of CL for the final model development. The developmental pharmacokinetic model was evaluated by goodness-of-fit plots and bootstrap analysis. The mean and variance of NPDE were -0.118 and 0.93, respectively. Conclusions: The effectiveness and pharmacokinetics were evaluated for the first time in pediatricAPL.The developmental population pharmacokinetic model for RIF in children revealed that one-compartment model with first-order elimination fitted the data.The current dosing regimen of 60 mg/kg/day TID resulted higher steady-state through concentration than that in adults (24.4μg/L, range:11.5 to 64μg/L). Disclosures:No relevant conflicts of interest to declare. This trial was conducted in accordance with the Declaration of Helsinki. OffLabel Disclosure: The combination of all-trans-retinoic acid (ATRA) and arsenic trioxide (ATO) has become the front-line treatment for patients with acute promyelocytic leukemia (APL). Realgar-Indigo Naturalis Formula (RIF), the only commercially oral arsenic agent, was launched in China from 2009. Since then, RIF had been used for over 5000 adults with acute promyelocytic leukemia (APL). One pill of RIF is 270 mg which contains 30 mg of Realgar, 125 mg of Indigo naturalis, 50 mg of Radix salviae miltiorrhizae, 45 mg of Radix pseudostellariae, and 20mg of garment film. However, the pharmacokinetics of RIF in children has not been studied.

The Electronic Flight Bag (EFB) provides pilots real-time or existing data by converting the papery documents into electronic books. Consequently the efficiency and the security have been improved, which reduce pilots’ ground communication time and the paperweight. As a key factor of what influences the takeoff process, the takeoff performance undoubtedly has been an essential research task of EFB, while many factors may affect the takeoff performance, such as flap position, anti-ice and the external temperature. By means of interpolation, this paper integrates those factors and gets Maximum Take-off Weight (TOW), Take-off Decision Speed (V1), Take-off Rotation Speed (VR) and Take-off Safety Speed (V2). And then implants the result into the EFB based on the iOS.

The major virulence determinant of Legionella pneumophila is the type IVB secretion system (T4BSS), which delivers approximately 330 effector proteins into the host cell to modulate various cellular processes. However, the functions of most effector proteins remain unclear. WipA, an effector, was the first phosphotyrosine phosphatase of Legionella with unknown function. In this study, we found that WipA induced relatively strong growth defects in yeast in a phosphatase activity-dependent manner. Phosphoproteomics data showed that WipA was likely involved into endocytosis, FcγR-mediated phagocytosis, tight junction, and regulation of actin cytoskeleton pathways. Western blotting further confirmed WipA dephosphorylates several proteins associated with actin polymerisation, such as p-N-WASP, p-ARP3, p-ACK1, and p-NCK1. Thus, we hypothesised that WipA targets N-WASP/ARP2/3 complex signalling pathway, leading to disturbance of actin polymerisation. Indeed, we demonstrated that WipA inhibits host F-actin polymerisation by reducing the G-actin to F-actin transition during L. penumophila infection. Furthermore, the intracellular proliferation of wipA/legK2 double mutant was significantly impaired at the late stage of infection, although the absence of WipA does not confer any further effect on actin polymerisation to the legK2 mutant. Collectively, this study provides unique insights into the WipA-mediated regulation of host actin polymerisation and assists us to elucidate the pathogenic mechanisms of L. pnuemophila infection.

Background The opportunistic bacterial pathogen Legionella pneumophila uses substrate effectors of Dot/Icm type IVB secretion system (T4BSS) to accomplish survival and replication in amoebae cells and mammalian alveolar macrophages. During the conversion between its highly resistant, infectious dormant form and vigorously growing, uninfectious replicative form, L. pneumophila utilizes a complicated regulatory network in which proteolysis may play a significant role. As a highly conserved core protease, ClpP is involved in various cellular processes as well as virulence in bacteria, and has been proved to be required for the expression of transmission traits and cell division of L. pneumophila. Results The clpP-deficient L. pneumophila strain failed to replicate and was digested in the first 3 h post-infection in mammalian cells J774A.1. Further investigation demonstrates that the clpP deficient mutant strain was unable to escape the endosome-lysosomal pathway in host cells. We also found that the clpP deficient mutant strain still expresses T4BSS components, induces contact-dependent cytotoxicity and translocate effector proteins RalF and LegK2, indicating that its T4BSS was overall functional. Interestingly, we further found that the translocation of several effector proteins is significantly reduced without ClpP. Conclusions The data indicate that ClpP plays an important role in regulating the virulence and effector translocation of Legionella pneumophila. Electronic supplementary material The online version of this article (doi:10.1186/s12866-016-0790-8) contains supplementary material, which is available to authorized users.

Fetus in fetu (FIF) is a very rare congenital malformation with an incidence of 1/500,000 live births. These children are often hospitalized because of vomiting, abdominal distension or abdominal mass found by their parents. The most common position for the parasitic fetus is the retroperitoneum but other position such as the scrotal sac, cranial cavity, sacrococcygeal region, back , oral cavity has been reported. FIF is often misdiagnosed as a mature teratoma. Ultrasound, CT, MRI are of great significance in distinguishing between FIF and teratoma. The postoperative pathological examination may reveal axial skeleton which can make a definitive diagnosis of FIF. In this report, we present a case of FIF in an 8-month-old girl.

To explore the sensitization effect of allergen TAT-IhC-R8, derived from major allergen group 1 genes of dust mites.Forty BALB/c mice were randomly divided into 4 groups, namely PBS group, ovalbumin (OVA) group, R8 group and TAT-IhC-R8 (TIR8) group, 10 mice each group. All the mice in OVA, R8 and TIR8 groups were treated with corresponding allergens (10 µg/ml) on the 0, 7th and 14th day by intraperitoneal injection and nebulized inhalation on day 21 with the concentration of 30 min/d for 7 days. The mice in PBS group were treated with PBS. Twenty-four hours after the last challenge, all the mice were sacrificed, their bronchoalveolar lavage fluids (BALFs) and sera were collected and their spleen cells were cultured. ELISA was performed to detect the levels of IFN-γ and IL-13 in BALFs and supernatants of cultured splenocytes (SCSs) of the mice, as well as the levels of allergen-specific IgE (sIgE), IgG, and IgG2 in their sera. The number of white blood cells and eosinophils in BALF were calculated. In addition, the airway inflammation and mucus secretion were analyzed by haematoxylin and eosin (HE) staining.Compared with the PBS group, the lung inflammations of mice in the OVA, R8 and TIR8 groups were observed obviously, including inflammatory infiltration, bronchial epithelial cell breakage and falling off, as well as vasculitis. The numbers of the total white blood cells and eosinophils in BALF of mice in the TIR8 group were significantly more than those in the OVA and R8 groups (all P0.01). The IL-13 levels in BALFs and SCSs of mice in the TIR8 group were significantly higher than those in the OVA group and R8 group (all P0.01). However, the level of IFN-γ of mice in the TIR8 group was lower than those in the latter 2 groups (all P0.01). In addition, the levels of sera sIgE and IgG of mice from the TIR8 group were significantly higher than those in the OVA group and R8 group (all P0.01), but the level of IgG2a of the former was significantly lower than those of the latter groups (all P0.01).TAT-IhC-R8 can effectively stimulate lung inflammations of mice, and its sensitization effect is better than R8's.

Soft tissue expansion is a technique useful in reconstruction when a shortage of tissue exists. This study presents an investigation on using chronic balloon expansion in the urethral and vaginal reconstruction of high persistent cloaca. The common channel was expanded by a balloon, which was progressively inflated over 3–4 weeks at a pressure of 30–50 mmHg until an adequate amount of tissue was obtained. The expanded channel was longitudinally split into anterior and posterior segments: the former was approximated to form a neourethra in continuity with the reconstructed bladder neck and the latter was tubularized to form a vagina. Twenty-one patients underwent primary posterior sagittal urethro-vaginoplasty using the expanded urogenital sinus. Histologic examination demonstrated that the expanded tissue consisted of a dense fibrous wall lined with a stratified squamous non-cornified epithelium, which was characterized by active cell mitosis and angiogenesis. The patients were followed-up for 0.6–9 year (mean 5.69 ± 2.43 years). All reconstructed urethras, vaginas, and vulvas were satisfactory in color, texture match, and sensation. Urinary continence (grades I and II) was observed in all patients without intermittent catheterization. Two complications were encountered in our study. One patient showed a distal urethral dehiscence and a secondary urethroplasty was performed 6 months after the operation. Another patient developed a redundant urethra upon reaching puberty and urethroplasty was needed. Chronic balloon expansion allows the formation of adequate tissue similar in appearance and type to the vagina and urethra. It also provides a valuable surgical alternative for the management of high persistent cloaca.

The mRNA level of the endonuclease domain-containing 1 gene (Jf_ENDOD1) in Japanese flounder Paralichthys olivaceus kidney was significantly increased after injection of formalin-killed bacteria cells (FKC) in the previous microarray study. ENDOD1 is a member of the DNA/RNA non-specific nucleases family, and its role in fish immunity has not been reported. The open reading frame of Jf_ENDOD1 cDNA was 912 bp, encoding 303 amino acids. The first 27 amino acids were predicted to be a signal peptide and the mature Jf_ENDOD1 was calculated as 32 kDa. The amino acid sequence of Jf_ENDOD1 showed 76% identity to that of large yellow croaker Larimichthys crocea. Transcripts of Jf_ENDOD1 were marginally detected in all sampled tissues from healthy fish, while they were significantly detected in brain, kidney, spleen and intestine at 6 h post FKC injection. Jf_ENDOD1 recombinant protein produced in Escherichia coli showed DNase activity. Furthermore, to evaluate the DNase activities in vivo, total proteins from Japanese flounder kidney and spleen were extracted at 12, 24 and 72 h post Edwardsiella tarda FKC injection. The DNase activity of extracted protein was higher in treated fish than in untreated fish. Since the mRNA levels were significantly up-regulated after the FKC treatment, Jf_ENDOD1 might be responsible for the activities.

A new type of CXCL8, named CXCL8_L1b, was identified in this research. Comparison of amino acid sequences of Japanese flounder CXCL8_L1b and CXCL8_L1a (BAB86884.1) showed only 41.2% identity. Transcripts of CXCL8_L1a were highly detected in spleen, kidney, gill and liver, while transcripts of CXCL8_L1b only were detected highly in spleen and kidney of apparently healthy fish. In fish challenged with E. tarda, transcripts of CXCL8_L1a were significantly increased at day 6, while no significant increase was detected in the mRNA level of CXCL8_L1b. On the other hand, fish infected by S. iniae significantly increased both transcripts of CXCL8_L1a and CXCL8_L1b at days 1 and 3. In VHSV-infected fish, only the transcripts of CXCL8_L1b were significantly induced at day 6. LPS and poly I:C stimulation of PBLs induced a high level of CXCL8_L1a transcripts, while CXCL8_L1b transcripts were significantly increased only post poly I:C treatment. To evaluate the chemotactic activity of CXCL8_L1a and CXCL8_L1b, Japanese flounder were intramuscularly injected with recombinant plasmids pCI-CXCL8_L1a and pCI-CXCL8_L1b. H & E staining showed that injections of both pCI-CXCL8_L1a and pCI-CXCL8_L1b caused strong immune responses in the form of intermuscular cell infiltration and capillary congestion. Injection of pCI-CXCL8_L1a and pCI-CXCL8_L1b significantly induced the expressions of genes related to inflammatory response such as IL-6 and CD8α on day 1 post-injection. The transcripts of IgM only significantly increased on day 7 post-injection of pCI-CXCL8_L1b.

To explore the effect of specific immunotherapy with major 3 group recombinant allergen rDer f 3 of Dermatophagoides farinae in murine asthma model.Forty BALB/c mice were randomly divided into 4 groups, namely PBS group (negative control), ovalbumin(0VA) group (positive control), rDerf3 allergen sensitization group (asthma group), and rDerf3 specific immunotherapy group(SIT group). The mice in asthma group and SIT group were injected intraperitoneally with purified rDer f 3 protein on days 0, 7 and 14, respectively, and rDer f 3 solution was inhalated from day 21 for 7 days. During the 25th-27th day, mice in SIT group were injected subcutaneously with 100 jg rDer f 3 allergen for 30 min before nasal inhalation. Mice in groups of PBS and OVA were treated with PBS and OVA, respectively. Twenty-four hours after the final challenge, all mice were sacrificed, the bronchoalveolar lavage fluid (BALF) was collected and the total number of white blood cells and the number of eosinophils were recorded. The levels of IL-5 and IFN-gamma in BALF and supernatant of cultured splenocytes were detected by ELISA, as well as the serum levels of specific IgE and IgG2, antibodies. Lung tissues were stained with haematoxylin and eosin for histological analysis.Compared with the asthma group, the rDer f 3-induced lung inflammation was significantly alleviated in SIT group. The total number of white blood cells [(7.03 +/- 1.38) x 10(8)/ml] in SIT group was considerably lower than that of OVA group [(22.11 +/- 3.70) x 10(8)/ml] and asthma group [(22.75 +/- 3.24) x 10(8)/ml] (P0.01). The change trend of eosinophil leukocytes was similar with that of white blood cells. IL-5 levels in BALF [(108.20 +/- 11.02) pg/ml] and splenocyte culture supernatant [(98.34 +/- 13.06) pg/ml] in SIT group were significantly lower than that of OVA group [(182.04 +/- 13.94) pg/ml, (208.26 +/- 10.63) pg/ml] and asthma group [(195.33 +/- 15.33) pg/mL, (179.54 +/- 13.65) pg/ml] (P0.01). Whereas, the level of IFN-gamma in BALF [(107.98 +/- 12.64) pg/ml] and supernatant of cultured splenocytes [(105.51 +/- 1.62) pg/ml] in SIT group was significantly higher than those of OVA group and asthma group (P0.01). Compared with OVA group [(26.87 +/- 4.30) IU/ml] and asthma group [(35.25 +/- 8.84) IU/ml], a lower level of allergen-specific IgE [(9.12 +/- 3.78) IU/ml] and higher level of allergen-specific IgG2, [(38.52 +/- 6.33) microg/ml] were observed in SIT group (P0.01).rDer f 3 allergen can reverse allergen-induced airway and lung inflammation in murine asthma model.

To predict and identify T cell epitopes of major group 3 allergen derived from Dermatophagoidesfarina (Der f 3).The T cell epitopes of Der f 3 were analyzed through the sequence analysis by using the bioinformatics online tools. The five predicted peptides of T-cell epitopes were artificially synthesized. The spleen lymphocytes were co-cultured with the five T cell epitopes by using the modified MTT method and the levels of IL-2, IFN-γ, IL-4 and IL-5 in the supernatant of the cultures were detected by ELISA.Five T cell epitopes of Der f 3 were predicted and three of which could promote the proliferation of the mouse spleen lymphocytes. The secretions of IL-2 and IFN-γ were significantly induced and the secretions of IL-4 and IL-5 were significantly decreased by three of five prediction epitopes of Der f 3: 37GDCPYQISLQSSSHFCGG54, 98IYQHENYDSMTIDNDVALIKLKTPMT123 and 164SELQRVDIDVVSREQCDQLYS184.Three T cell epitopes of Der f 3 have been initially identified, which lays the foundation of the diagnosis and treatment of asthma.

To predict and identify the linear B-cell epitopes in the major group 3 allergen derived from Dermatophagoides farina (Der f 3).The linear B-cell epitopes of Der f 3 allergen were analyzed based on the physicochemical properties of amino acids including antigenicity, surface accessibility, flexibility, hydrophilicity, beta-turn by online bioinformatics softwares. The eight predicted peptides of linear B-cell epitopes were artificially synthesized and incubated with three aller gic serum pools (4 serum samples in each), which were consisted of total 12 serum samples from the allergic individuals, and the strong positive epitopes were selected.Eight B-cell epitopes from Der f 3 were predicted successfully. Five of eight B-cell epitopes were identified with strong IgE-binding abilities followed by specific IgE assay. The amino acid sequences of them were following: 33KAKAGDCP40, 86HASGGEKIQVAEIYQHENYDSMTID110, 118LKTPMTLDQTNAKPVPLPPQGSDVKVG144, 156QEGSYSLP163 and 199DVANGGVDSCQGDSGGPVVD218.Five linear B-cell epitopes of Der f 3 allergen have been identified successfully. This result might provide a basis of the diagnosis and treatment for asthma.

Specific immunotherapy (SIT) is the only treatment that provides long lasting relief of allergy symptoms. Unfortunately, SIT-based traditional remedies carry the risk of producing local and/or systemic side effects. To improve the safety and efficacy of SIT, it has been proposed that SIT must utilize allergens that are hypoallergenic but hyperimmunogenic. Therefore, we used DNA shuffling to generate mutant genes encoding hypoallergens with potent immunogenicity and screened them for their capacity to modify the allergic response. We tentatively shuffled the major group 1 allergen genes from house dust mite, Dermatophagoides farinae and Dermatophagoides pteronyssinus, and discovered a novel chimeric gene, termed C 1. The gene was expressed in Escherichia coli (E. coli) and the chimeric protein C 1 was purified. An animal model of asthma demonstrated that C 1 not only decreased the production of serum IgE and IgG1, and inhibited the production of IL-4 and IL-5 in the bronchoalveolar lavage fluid (BALF). C 1 also boosted the levels of IgG2a and IFN-γ, which may demonstrate a rebalance of TH1 and TH2 allergic response. Additionally, flow cytometry showed that the immunogenicity of C 1 was higher than that of ProDer f 1, but was not significantly different from that of ProDer p 1. Our findings suggest that the C 1 is hypoallergenic and yet highly immunogenic, which makes it potentially safe and effective for use in SIT of allergic asthma.