Alternative splicing of pre-messenger RNA (pre-mRNA) is a crucial mechanism for the diversity of the human transcriptome and proteome. Alternative splicing is a complex gene regulation process. Whole-transcriptome analysis shows that 95% of human exonic genes are alternatively spliced, involving various cis-acting elements and trans-acting factors. Any changes in any component or step may cause erroneous splicing events and lead to the occurrence of various related diseases. In addition to gene replacement therapy that directly changes the splicing results, RNA splicing modification is expected to become a new therapeutic strategy to alleviate or treat diseases by targeting and correcting abnormal pre-mRNA splicing. Splicing modification tools currently developed including RNA trans-splicing, antisense oligonucleotides, small interfering RNA, and small molecule drugs can correct abnormal splicing through different ways. This article reviews the resent progress of epigenetic regulation of pre-mRNA alternative splicing in recent years, and discusses the occurrence and regulation of alternative splicing, the types of diseases caused by related splicing defects, and the current-used tools for targeting and altering splicing. The importance of splicing modification strategies in the future treatment of human diseases is envisioned.

Asphyxia Neonatorum ; blood ; drug therapy ; Creatine Kinase, MB Form ; blood ; Female ; Humans ; Infant, Newborn ; Male ; Naltrexone ; analogs & derivatives ; therapeutic use ; Natriuretic Peptide, Brain ; blood ; Peptide Fragments ; blood

objective To investigate the clinical signiifcance of the changes of angiotensionⅡ(AngⅡ) in children with IgA nephropathy (IgAN). Methods Thirty children diagnosed as primary IgA nephropathy by renal biopsy (IgAN group) and 30 healthy children (control group) were recruited from May 2008 to December 2012. The serum and urine AngⅡwere measured by ELISA and compared between IgAN group and control group. The AngⅡexpression in the renal tissue of IgAN group was detected by immuno-histochemical method, and was correlated with other clinical data.. Results Urine AngⅡwas signiifcantly higher in the primary IgAN group than that of control group (P<0.05);AngⅡexpression in the urine is positively correlated with proteinuria (r=0.37, P=0.046), and is associated with the severity of clinical presentation; AngⅡexpression in kidney tissue increased with the severity of the renal histopathologic grading (r=0.69, 0.79, P=0.000), while AngⅡin blood and proteinuria, AngⅡexpression in kidney tissue were not signiifcantly correlated with the number of crescents. Conclusions Urine AngⅡin children with IgAN is signiifcantly correlated with the severity of the pathologic stage and the level of proteinuria. Urine AngⅡdetection may be useful to assess the progress and prognosis of chronic kidney disease.

Objective To investigate the synergistic effect of methotrexate (MTX) and cyclophos phamide (CTX) on cell cycle and cyclin D1 of periphery blood lymphocytes (PBLs) and bone marrow byflow cytometry. Methods Wistar rats were randomly divided into four groups including normal control, MTX and cyclophosphamide combination group, MTX and CTX only treatment groups respectively. PBLs were isolated for flowcytometry analysis for the changes of cell cycle and the expression of cyclin D1 at week 0, week 3,week 9, week18 and week 27. Mice were dissected and the changes of lymphoeytes cell cycle and the expressions of cyclin D1 in the bone marrow were measured at week 0, week 3, week 9, week 18 and week 27 increased and the ratio of phase S cells was decreased (P>0.05). In the CTX treatment group, there was no statistical difference in ratios of each phase. In the MTX and CTX combination treatment group, the proportion of phase G0/G1 cells decreased significantly and the percentage of phase S cells increased in both PBLs and bone marrow ceils (P<0.05). And there was no statistical significant difference in different time points after marrow between different groups or different dissecting time points. Conclusion MTX combined with CTX has been shown to have antagonistic effect on cell cycle. However, this effect is not via the cyclin DI pathway.

To determine the etiologic agent of an outbreak of influenza viruses from Changsha Foothill Mountain International School in 2009, and to analyze the HA Gene Characteristic of the H1N1 influenza viruses. Twenty-five nasopharyngeal swab specimens from the outbreak of influenza viruses were tested by conventional RT-PCR and influenza viruses isolated simultaneously. Virus isolated (A/Yuelu/314/2009) from the outbreak was sequenced by CEQTM 8000 Genetic Analysis System and the sequencing results submitted to GenBank (Accession No: FJ912843), then the sequencing data was analyzed by ClustalX and Mega4.1softwares. Results showed the influenza viruses A(H1N1) of positive were 18 cases by influenza viruses isolated tests and 21 cases by conventional RT-PCR, respectively. The nucleotide and amino acid sequence homology of the HA gene of A/Yuelu/314/2009 are 99% compare with the vaccine strain (A/Brisbane/59/2007) in 2008~2009 years. The HA sequence data also showed that had 6 amino acid mutations (V148A, S158N, G202A, I203D, A206T, W435R), and the S158N located at antigenic site B of HA protein. Nine potential glycosylation sites (27, 28, 40, 71, 151, 176, 303, 497, 536) in the HA sequence of A/Yuelu/314/2009 is the same with A/Brisbane/59/2007, and the sequences of potential glycosylation sites were conserved. In this study, laboratory evidence diagnosed seasonal influenza A virus (H1N1) as the etiologic agent of the outbreak. The virus isolated (A/Yuelu/314/2009) strain of H1N1 subtype is not a new variant in Changsha in 2009 compare with the vaccine strain (A/Brisbane/59/2007), the outbreak of influenza A virus (H1N1) from Changsha Foothill Mountain International School maybe are caused by the change in genetic characteristics between vaccine strains and the decreased of immunity to influenza A virus (H1N1) in the crowd.

Objective To observe the pathological changes of bone hydatid cyst of meriones meridianus after radiation therapy,and to investigate the clinical effect of radiotherapy on bone hydatid disease.Methods Ascus was dissected sterilely from sheep liver that naturally infected with Echinococcus granulomas,sheared and sac skin removed.Then it was washed and precipitated with 0.9％ sterile saline for 3 times,and scolex was HE stained and counted,from which a 20 ml suspension was made containing 12 × 106/L of scolex.Health meriones meridianus (referred to as gerbil) 140,male and female were in each half,aged 2 to 3 months,body weight(38 ± 6)g,were involved in the study.Gerbil was injected a 0.2 ml suspension containing Echinococcus granulomas scolex into hind tibial periosteum,and X-ray was taken 12 months after the injection.According to the bone destruction in the vaccination site,gerbil hindleg tibia with clear jagged bone destruction was treated as inclusion criteria,and 72 animal were selected as gerbil bone hydatid disease animal models,male and female were in each half.A tatal of 72 gerbils were randomly divided into 4 groups:control group,40 beequerel(Gy) group,50 Gy group and 60 Gy group,18 rats in each group,male and female in each half.The model animals were treated with radiotherapy for 5 times,with 2 d interval,and radiation dose was 300 cGy/min.Each group of gerbils was sacrificed after radiotherapy,bone Echinococcus granulomas cysts was taken out sterilely,and observed by light and electron microscope.Intracapsular cyst fluid was extracted,washed and precipitated with 0.9％ sterile saline repeatedly,and and the pellet was HE stained for observation of scolex morphology and activity by light microscope.Results The morphology and activity ofEchinococcus granulomas in cystic fluid in control group were normal; the morphology and activity of Echinococcus granulomas were still normal in the 40 Gy group,and Echinococcus granulomas was not stained red; but those were abnormal,deformation and atrophy and stained red in the 50 Gy group; and were stained red,deformed,fractured and were wrapped by unknown in the 60 Gy group.By light microscope,the germinal layer,cuticle layer,brood capsule and histological structure of protoscolex were basically normal in irradiated region in the control group.The pathological changes of hydatid cyst in the 40 Gy group were mainly degeneration,structure of hydatid cyst was abnormal,stratum corneum was extensive edema,germinal layer became thinner and the fertile cyst was rare.The main pathological change of hydatid cyst in the 50 Gy group was that corneous layer was widely fractured,and the germinal layer was edema,buckling folds,cells decreased,rare seen brood capsule and scolex; the main pathological changes of hydatid cyst were mainly necrosis in the 60 Gy group,cuticle was extensive fault,stratum corneum and germinal layer was separated,germinal layer was atrophy and disorder,no brood capsule and scolex.By electron microscope,cuticle structure of Echinococcus granulomas cyst was clear,microvillus arranged neatly,morphology and structure of the cell and organelle in cytoplasm were normal in the control group.There were many inflammatory cells infiltrating germinal layer of Echinococcus granulomas cyst,microfilament and contents in microfilament were reduced in the 40 Gy group.Microvillus of Echinococcus granulomas disappeared,nuclear membrane was unclear,endoplasmic and mitochon eclasis,lymphocyte nuclear chromatin was clumping and edge set and in circular permutation in the 50 Gy group.Microvillus disappeared,perinuclear membrane indistinct and ruptured,parts of nucleoli were fragmented and marinated,endoplasmic reticulum was extensive expansion,mitochondria was pyknosis and obvious vacuolization,lymphocyte nuclear chromatin clumping and edge set,lysosomes and macrophage emerge in the 60 Gy group.Conclusions Radiotherapy can destroy the morphology and structure of bone hydatid cyst,radioactivity at 50 Gy has a lethal effect on hydatid cyst.Radiation treatment of bone hydatid disease has a good clinical effect.

Objective To evaluate the clinical impact of 18F-FDG PET/CT on patients with suspected cervical cancer recurrence. Methods Fifty-one cervical cancer patients, clinically suspected to have tumor recurrence during follow-up, underwent 18F-FDG PET/CT examination. 18 F-FDG PET/CT results were compared with those of conventional images, as referred to histopathology or clinical follow-up. Impacts of 18F-FDG PET/CT were evaluated based on documented changes of clinical management. Results In total, 43 patients were found to have positive lesions by 18F-FDG PET/CT, in which 40 were true recurrence,but 2 were pelvic abscess and 1 was radiation enterocolitis. Other 8 patients were found negative by 18F-FDG PET/CT and confirmed by pathology or follow-up. In patient-based analyses, the sensitivity, specificity, and accuracy of 18F-FDG PET/CT for the detection of tumor recurrence were 100% (40/40), 72. 73% (8/11),and 94.12% (48/51) respectively. In 7 patients, the clinical management was changed due to 18F-FDG PET/CT findings. Conclusion 18F-FDG PET/CT is an efficient tool for determining the recurrence of cervical cancer and instructing the clinical management.

AIM:To establish the GC fingerprint of Huoxiang Zhengqi Solution.METHODS:The volatile constituents of Huoxiang Zhengqi Solution were analyzed by capillary GC with FID detector using hydrodistillation and hexane extraction under n-hepladecane used as the reference substance.RESULTS:GC fingerprint of Huoxiang Zhengqi Solution,16 common peaks were established on the basis of systematic methodology after 10 batches of samples were tested.Variation in the relative retention time of 16 identified common peaks were within 0.5% range.CONCLUSION:The analytical method for Huoxiang Zhengqi Solution is precise and reliable.The research would be helpful to offer an effective pattern for quality control of Huoxiang Zhengqi Solution.

AIM:To establish the GC fingerprint of Huoxiang Zhengqi Solution.METHODS:The volatile constituents of Huoxiang Zhengqi Solution were analyzed by capillary GC with FID detector using hydrodistillation and hexane extraction under n-heptadecane used as the reference substance.RESULTS:GC fingerprint of Huoxiang Zhengqi Solution,16 common peaks were established on the basis of systematic methodology after 10 batches of samples were tested.Variation in the relative retention time of 16 identified common peaks were within 0.5% range.CONCLUSION:The analytical method for Huoxiang Zhengqi Solution is precise and reliable.The research would be helpful to offer an effective pattern for quality control of Huoxiang Zhengqi Solution.

To explore the action target of praziquantel (PZQ) and its underlying mechanism of action, adult male worms of Schistosoma japonicum were collected from the hepatic vein of Kunming mice infected at least 6 or more weeks previously with single-sex cercariae of S.japonicum by perfusion method. These worms were subjected to the action of calcium channel blockers (CCBs) or actin depolymerizing/stabilizing agents interfering with function of the calcium channel. The adult male worms in DMEM culture medium were co-cultivated with near-lethal dose of PZQ(14 μmol/L) overnight(16 hours).Then, the parasites were washed 3 times with sterile physiologic saline next morning after cultivation, re-suspended in new and drug-free medium and then observed under stereo-microscopy during the following 5 days. The experimental results showed that majority of adult male worms of S.japonicum were killed by the action of PZQ in a dose of 14 μmol/L in vitro under normal condition; while the worms pre-incubated with the actin depolymerizing agent cyto chalasin D (CyD) were able to survive in the condition containing 14 μmol/L of PZQ with a survival rate of 100%, and the worms pre-incubted with CCBs, such as nitrendipine and nifedipineu showed a survival rate of about 50% under the same condition. The results of this study suggest that the calcium channel of Schistosomes may be involved in the action target of PZQ and its underlying mechanism.