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 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.

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.

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.

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.

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.

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.

Objective To evaluate the effectiveness of maternal prenatal education on promoting breastfeeding rate for preterm infants in neonatal intensive care unit (NICU).Methods Gravidas who were admitted to Peking University First Hospital for any risk of preterm delivery from November 2013 to December 2014 and preterm infants admitted to the NICU (length of stay ≥ 5 d) of the same hospital during the same period were involved.All the gravidas received prenatal education and completed questionnaires.Two hundred and ninety-five preterm infants who met the inclusion criteria were divided into two groups including education group (n=125) and non-education group (n=170) according to whether their mothers received prenatal education or not.Conditions of the preterm infants during hospitalization and after discharge and breastfeeding rates were comparatively analyzed between the two groups.T test,Chi-test and Wilcoxon rank-sum test were used for statistical analysis.Results (1) There were 380 gravidas received questionnaires.Among them,346 (91.1％)completed the questionnaires and were recruited in this study.Only 31.8％ (110/346) of these gravidas were active in learning more about breastfeeding and 46.2％ (160/346) of them lacked confidence in breastfeeding after the appearance of preterm delivery signs.There were significant improvements in their attitudes towards considering breastfeeding seriously and discussing with their family,confidence in breastfeeding,the importance of colostrum and how to breastfeed a preterm infant in hospital after maternal prenatal education (all P＜0.001).Prenatal education was thought to be helpful in 77.5％ (268/346) of the gravidas.(2) The two groups of preterm infants showed good homogeneities in gestational age,gender,birth weight and other basal conditions as well as in incidences of in-hospital complications and conditions after discharge (all P＞0.05).Proportions of breast milk intake (breast milk intake over total dairy intake) in preterm infants were higher in education group than those in non-education group within 5 d after birth [0.0 (0.0-16.5)％ vs 0.0 (0.0-2.5)％,Z=-3.422],＞5-≤ 7 d [33.7 (0.0-82.8)％ vs 0.0 (0.0-50.3)％,Z=-3.070],＞7-≤ 14 d [75.2(23.5-96.4)％ vs 47.6(0.0-92.2)％,Z=-2.345] and during hospitalization [58.4 (21.0-78.8)％ vs 31.9 (0.0-71.7)％,Z=-3.902] (all P＜0.05).Breastfeeding rates were higher in education group than those in non-education group at the age of 5 d [47.2％(59/125) vs 27.1％ (46/170),x2=12.747],7 d [70.4％ (88/125) vs 51.2％ (87/170),x2=11.031],three months [83.3％(65/78) vs 56.1％ (60/107),x2=15.297] and six months [64.5％ (49/76) vs 49.1％ (53/108),x2=4.282] (all P＜0.05).Exclusive breastfeeding rates in the first,third and sixth months after birth were higher in education group [45.7％(53/116),42.3％ (33/78) and 36.9％ (28/76)] than those in non-education group [21.3％ (32/150),28.0％ (30/107)and 22.2％ (24/108)] (22=17.847,4.091 and 4.703,all P＜0.05),respectively.Conclusions Most gravidas with risk factors of preterm delivery have no confidence on breastfeeding.Prenatal maternal education is an effective and feasible intervention to improve breastfeeding rate for preterm infants in NICU from early hospitalization till six months after birth.

Objective To study the clinical presentations,radiologic features,prognosis,and possible causes of liver injury associated umbilical venous catheter (UVC ).Methods We reviewed database of our NICU from December 2012 to November 2015 and identified preterm infants with liver injury while UVC in place.The gestational age,birth weight,gender,days of UVC in place prior to liver injury, the depth of UVC (cm),UVC tip position,ultrasound findings of liver injury,laboratory tests,treatment and outcomes were collected.Results During study period,322 infants received UVC insertion.Ten cases (3.1 %)of liver injury associated with UVC were diagnosed.Of the ten infants,nine were diagnosed as extravasation of fluid to liver parenchyma,and one was diagnosed as liver hematoma.The mean birth weight was (1184 ±207)g,mean gestational age was (28.5 ±1.7)weeks.The UVC tip was at thoracic (T) vertebrae T 9 to T 11 level,nine UVC tips were below the right diaphragm level.All UVCs were used for total parenteral nutrition before liver injury was diagnosed.Six infants had clinical manifestations,and four infants were free of clinical manifestations.The most prominent clinical signs were abdominal distension (n =5),weak bowel sounds (n =5)and hepatomegaly (n =4).One infant who was diagnosed with liver hematoma also had progressive hemoglobin decline (minimum 34 g/L)and shock.Abdominal ultrasound showed well-limited,irregular,hyperechoic rimmed lesions with heterogeneously hypoechoic centers or anechoic liquid dark space.Extravasation of fluid to liver parenchyma will restorate within one week after the UVC was remove.One baby who was diagnosed as liver hematoma passed away at 9 days of life.Liver ultrasound of eight returned to normal in 52 days to 3.5 months,and one had calcified lesions at 9 months of age.Conclusions Liver injury is an unusual complication of UVC insertion and usage.Proper positioning of the UVC tip may help to avoid this complication.Early recognition,prompt diagnosis with liver ultrasound examination and timely treatment can lead to better outcome in newborns with extravasation of fluid to liver parenchyma.