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.

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 causes, clinical features, treatment and prognosis of gastrointestinal bleeding (GIB) patients in emergency department.Methods To analyze prospectively the clinical data of 168 GIB patients admitted to the emergency department of Peking Union Medical College Hospital during 2006.1-2006.12.Results (1) General data; male: female = 1.75:1 ( 107: 61) , mean age 13-87(56.5 ±17.8) years with a peak in 60-69 years.The percentage of old patients was significantly higher than that of young and middle age ( 52.4% vs 19.6% and 28.0% , P = 0.000 ).( 2 ) The incidence of acute gastric mucosal lesion in patients taking non-steroidal antiinflammatory drugs ( NSAIDs) ( 18.5% ) was significantly higher than that in patients not taking( 0.7% , P = 0.000 ).( 3 ) 86.9% ( 146/168 ) of the patients had anemia.(4) More patients who took emergency gastroscopy could be diagnosed than those patients who did not (89.4% vs 58.5% , P =0.000), while no significant difference could be seen between patients who took emergency enteroscopy and patients who had non-emergency gastroscopy (20.0% vs 57.9% , P =0.315).(5)The hemostatic ratio in GIB patients due to peptic ulcer was obviously higher than that in GIB patients due to other causes (86.0% vs 40.7% ,P =0.000).The rate of emergency operation for GIB patients was 1.8%.Conclusions Most of the GIB patients admitted to tertiary general hospitals are elderly males.NSAIDs administration is one of the most important causes of upper GIB.Upper GIB patients should have gastroscopy as soon as possible, while emergency coloscopy is of little significance in cases with lower gastrointestinal hemorrhage.

OBJECTIVETo analyzed the relationship between lumbar endplate Modic area changes rate and low back pain by measuring MRI T2 sagittal image of lumbar endplate Modic area changes rate.

METHODSFrom December 2011 to June 2012,70 patients with low back pain in operation were evaluated on pain by VAS and function by JOA,and examined by MRI including 39 males and 31 females with an average age of (51.00 +/- 11.89) years ranging from 29 to 72 years old. Among them, 54 cases had lumbar endplate Modic changes involving 15 cases in types Modic I ,21 cases in type Modic II, 11 cases in type Modic III ,mixed type Modic in 7 cases (eliminated for too few cases). Modic area changes and corresponding vertebral area were measured on MRI T2 median sagittal. The areas of two ways were compared to yield the rate of changes for Modic, for multisegmental Modic changes to calculate the total ratios. A correlation was observed among JOA, VAS and the rate of Modic changes.

RESULTSThe correlation coefficient of change rate of Modic I with JOA score was r = -0.308, P = 0.048 < 0.05, there was a negative correlation;the correlation coefficient of change rate of Modic I with VAS scores was r = 0.428,P = 0.021 < 0.05, there was a positive correlation. The correlation coefficient of change rate of Modic II with JOA score was r = -0.375, P = 0.043 < 0.05, there was a negative correlation;the correlation coefficient of change rate of Modic II with VAS score was r = 0.352, P = 0.041 < 0.05, there was a positive correlation. The area change rate of Modic III had no significant correlation with low back pain degree (P > 0.05).

CONCLUSIONModic I and II area changes rate of of patients with low back pain is closely related to the degree of pain low back pain, Modic III area changes rate is not significant correlated to the degree of lower back pain.

Adult ; Aged ; Female ; Humans ; Low Back Pain ; diagnosis ; diagnostic imaging ; Lumbar Vertebrae ; diagnostic imaging ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Radiography

Objective To explore the changes serum S-100? in children with acute carbon monoxide poisoning and its clinical significance.Methods The levels of serum S-100? of 28 children with acute carbon monoxide poisoning and those of 20 healthy children were mea-sured by enzyme-linked immunosorbent assay.Results The serum S-100? levels of the study group and control group were(0.517?0.346)and(0.037?0.014)?g/L respectively,there was significant difference between two groups(t=6.197 P

Purpose To investigate the expression and significance of androgen receptor (AR) in breast carcinoma with different ER and PR status.Methods Immunohistochemical method was used to detect AR, ER and PR expression in 173 cases of breast carcinoma, and all the cases were grouped according to: (1) AR status: AR~- positive subgroup and AR~- negative subgroup, (2) ER status: En subgroup (negative for both ER and PR), Ep subgroup (positive for ER and/or PR), (3)AR, ER and PR status: En-AR~+ subgroup (AR~- positive cases of En subgroup), En-AR~- subgroup (AR~-negative cases of En subgroup), Ep-AR~+ subgroup (AR~- positive cases of Ep subgroup), and Ep-AR~- subgroup (AR~-negative cases of Ep subgroup).En-AR~- subgroup also was called negativE-for-all subgroup, and the rest three subgroups were called partly-or-completely positive subgroup (positive for at least one of the three markers), and then, the groups were compared with clinicopathological features.Results The positive rate of AR was 62.8% (54/86) in Ep subgroup and 37.9%(33/87) in En subgroup, and there was significant difference between them (P=0.001).Compared to AR~-negative subgroup, AR~-positive subgroup had smaller tumor size, less mitosis count and lower grade(P<0.05).Compared to the other three subgroups, En-AR~- subgroup had more mitosis count and higher grade (P<0.01).In En subgroup,AR~- positive cases had less mitosis count and lower grade (P<0.05), and in Ep subgroup,AR~- positive cases had higher stage(P=0.000), but no significant difference were found among partly or completely positive subgroups.Conclusions The expression of AR may play a different role in breast carcinoma with different ER and PR status, because it indicates good prognosis in negative for both ER and PR subgroup, but bad in ER and/or PR positive subgroup. When choosing a personalized therapy, all the combinations of hormone receptor status should be considered for the patients.

Objective This study is designed to explore the regulation of carboxyamidotriazole(CAI)on inflammatory factors in vitro and its underlying mechanisms.Methods Peritoneal macrophages were incubated with different concentrations(5～40 μmol/L),and then cell viabilities were evaluated by MTT assay.Cells were pretreated with CAI(5～40 μmol/L),LPS was then added,and celia were incubated for 18 h.NO and TNF-α levels were determined with Griess reagent and ELISA kit,respectively.The iNOS expression and NF-κB activation were detected by Western blot method.Results The rat peritoneal macrophage viability was not affected at the concentrations of CAI used.CAI(5～40 μmol/L)was found to reduce NO(P＜0.01,P＜0.001)and TNF-α production(P＜0.05,P ＜ 0.01)in a dose-dependent manner.CAI was also found to inhibit the LPS-induced expression of iNOS and degradation of IκBα in rat peritoneal macrophages.Conclusion The findings from the present study suggest that CAI has suppressive effect on iNOS expression,and this inhibitory effect was found to be associated with NF-κB inactivation via the blockade of IκBα phosphorylation.

HIV p24 core protein can induce both cellular and neutralizing antibody responses.HIV-1 CA-virus-like particles(VLPs)vaccines provide a promising approach for the development of an effective vaccination strategy against HIV infection.Rhizosecreion of the recombinant proteins provides a new manufacturing platform that can simplify the extraction and purification procedure.Lycium barbarum L.was transformed by Agrobacterium tumefaciens EHA105 harboring the plant expression vector pCAMBIA1305.2-MA4-CA with a GRP signal peptide and MA4-CA fusion gene.Transgenic hairy roots were induced and cultivated in hydroponic culture.Western blotting indicated that the recombinant CA proteins were present in two forms,a glycosylated monomer(37 kDa)and a dimer(50 kDa)in the roots and hydroponic medium.It appeared from the present immunohistochemical data that the recombinant CA proteins fused with GRP signal peptide were confined to the cytoplasm,cell wall and intercellular space,indicating targeting into the secretory pathway.It demonstrated for the first time the rhizosecretion of HIV-1 recombinant capsid protein in Lycium barbarum L.hairy roots,and may offer a novel method for expressing HIV-1 CA-VLPs vaccines in plants.

Objective:To screen for the pathogenesis-related genes of osteosarcoma and to assess their roles for the de- velopment of osteosareoma.Methods:Total RNA was extracted from 3 ATCC osteosarcoma cell lines and an osteoblastic cell line and was used to synthesize biotinylated cRNAs;the latter were hybridized to Affymetrix~(?)GeneChip~(?)U133A ar- rays and a gene with more than 2 folds of change was selected.Ten of the differentially expressed genes were chosen and the primers were designed and the synthesized.Then SYBR~(?)Green real-time PCR(RT-PCR)method was used to detect the expression of the 10 genes in 9 fresh osteosarcoma specimens.ABI Prism 7 000 system was used to analyze the differ- ent expression between osteosarcoma cell line and osteoblastic cell line.Results:We identified 58 up-regulated and 142 down-regulated genes in the 3 osteosareoma cell lines.Many of the genes were firstly reported to be related to the patho- genesis of osteosarcoma.These differentially expressed genes were mainly involved in energy and material metabolism,on- cogene,signal transduction gene,transcription- related genes,cell cycle-related genes,cell apoptosis-related gene,im- mune response gene,tumor suppressor genes,etc.The array results of 10 randomly selected genes were further verified by the RT-PCR in 9 fresh osteosarcoma specimens.Conclusion:Many genes are involved in the pathogenesis of osteosarcoma. Gene microarray can help to discover the genes related to the pathogenesis of osteosarcoma,which may lay a foundation for studying the molecular mechanism of osteosarcom.

OBJECTIVE To investigate the transcription level of gene hld of Staphylococcus epidermidis in the biofilm forming and detaching under MIC antibiotic and explore the relationship between biofilm-related drug resistance and persistant infection.METHODS The transcription level of gene hld of S.epidermidis under MIC concentration of 4 antibiotics was compared with those of the control group without antibiotics by SYBR real-time fluorescent quantitative RT-PCR at the different time point of biofilm formation and detachment.RESULTS The transcription of gene hld decreased rapidly from initial adherence,and droped continuously for few hours.There was an increase from 24 hours to 72 hours in groups without antibiotics but not in antibiotics groups,the differenet was significant.CONCLUSIONS Antibiotics improve adherence at first and then prevent matrix decomposition water-conducting tube and detachment of cells by impact of transcription of gene hld,it can protect cells from killing by inhibiting the penetration of biotics and prevent them become planktonic cells after detachment from biofilm.