RNA therapeutics inhibit the expression of specific proteins/RNAs by targeting complementary sequences of corresponding genes or encode proteins for the synthesis desired genes to treat genetic diseases. RNA-based therapeutics are categorized as oligonucleotide drugs (antisense oligonucleotides, small interfering RNA, RNA aptamers), and mRNA drugs. The antisense oligonucleotides and small interfering RNA for treatment of genetic diseases have been approved by the FDA in the United States, while RNA aptamers and mRNA drugs are still in clinical trials. Chemical modifications can be applied to RNA drugs, such as pseudouridine modification of mRNA, to reduce immunogenicity and improve the efficacy. The secure and effective delivery systems such as lipid-based nanoparticles, extracellular vesicles, and virus-like particles are under development to address stability, specificity, and safety issues of RNA drugs. This article provides an overview of the specific molecular mechanisms of eleven RNA drugs currently used for treating genetic diseases, and discusses the research progress of chemical modifications and delivery systems of RNA drugs.
Aptamers, Nucleotide ; RNA, Small Interfering/therapeutic use* ; RNA, Messenger ; Oligonucleotides, Antisense/therapeutic use*

RNA therapeutics inhibit the expression of specific proteins/RNAs by targeting complementary sequences of corresponding genes, or synthesize proteins encoded by the desired genes to treat genetic diseases. RNA-based therapeutics are categorized as oligonucleotide drugs (antisense oligonucleotides, small interfering RNA, RNA aptamers), and mRNA drugs. The antisense oligonucleotides and small interfering RNA for treatment of genetic diseases have been approved by the FDA in the United State, while RNA aptamers and mRNA drugs are still in clinical trials. Chemical modifications are applied to RNA drugs, such as pseudouridine modification of mRNA, to reduce immunogenicity and improve the efficacy. The secure and effective delivery systems like lipid-based nanoparticles, extracellular vesicles, and virus-like particles are under development to address stability, specificity, and safety issues of RNA drugs. This article provides an overview of the specific molecular mechanisms of 11 RNA drugs currently used for treating genetic diseases, and discusses the research progress of chemical modifications and delivery systems of RNA drugs.

Objective To explore the layers of the facial and angular arteries between the orbital notch and the antegonial notch.Methods Anatomical studies were performed on 5 cases of adult cadavers (10 sides).Parallel lines were drawn on each cadaver through the position of the orbital incise,inner canthus,above of wing nose,nasal wing point,mouth angle,anterior incise.When the region of the arterial level changes,the parallel lines could be temporarily divided.The levels of the arteries that appeared on every horizontal line were recorded.Results The surface artery between the orbital notch (including the supraorbital notch) and the inner canthus (including the inner cantholes) was mainly in the orbicularis muscle layer.In the middle of the inner canthus and the highest point of the nasal wing,1/3 of the arteries were mainly walking in the orbicularis orbicularis muscular layer,and the next 1/3 were mainly located in the superficial layer or subcutaneous fat layer of the orbicularis oculi muscle.The highest point of the nose wing was found in the middle of the subcutaneous fat.The main line between the top of the nose and the lower part of the nose was deep in the subcutaneous fat.At the bottom of the nose,the main line was located in the fat layer of the zygomaticus,the zygomaticus maximus.In the middle and upper part of the nose and the middle of the mouth,a third of the main lines were walking in the lower fat layer (from shallow to deep) under the zygomaticus,and the next 1/3 segment was found in the subcutaneous fat deep.On the outside of the corner of the mouth,the main line was walking the subcutaneous fat layer or the lower fat layer of the platysma.Conclusions This study preliminarily clarifies the walking level of the facial artery from the upper orbital notch and the anterior notch of the angle.Bearing in mind the depth and the location of the vasculature within each zone,clinical practitioners can tailor their injection techniques to prevent vessel injury and avoid cannulation.

Objective To explore the clinical characteristics of children with dengue fever (DF) hospitalized in Guangzhou in 2014 , and to raise clinician′s level of understanding of dengue fever in children.Methods Clinical data of 78 children hospitalized with DF in the Department of Infectious Diseases ,Guangzhou Women and Children′s Medical Center were retrospectively analyzed .Chi-square was used for discontinuous variables ,and t test was used for continuous variabbles .Results The 78 cases aged 27 days to 14 years old ,with median of 5 years old .Fifty cases (64 .1%) were male ,and 28 cases (35 .9%) were female.Epidemic areas had gathering trend ,mainly in central urban area .Major clinical manifestations were fever (100 .00%) , rash (82 .05%) , myalgia／fatigue (28 .21%) , but without diarrhea ,jaundice ,hematemesis or hematochezia .Laboratory tests suggested leukopenia (80 .77%) and thrombocytopenia (82 .05%) ,abnormal blood coagulation function with prolonged APTT (57 .69%) ,and abnormal liver function (47 .44%).Etiology examinations showed 66 cases of children had dengue virus nucleic acid detected 1－10 days after onset ,with the positive rate of 89 .19%(66／74).A total of 48 cases had IgM positive ,with the positive rate of 81 .36%(48／59).IgM began to appear as early as the first day of disease onset ,and the average period was (5 .5 ± 0 .8) days .Dengue virus type 1 was the main type . Conclusions In 2014 , dengue virus type 1 is the main strain causing dengue fever in children in Guangzhou .Fever ,rash ,leukopenia ,thrombocytopenia ,clotting disorders and liver function damage are the main clinical features .No serious or fatal cases are reported ,and the prognosis is good.

Objective To detecte the neovascularizations in carotid plaques using contrast-enhanced ultrasound (CEUS) and explore the relationship between the neovascularizations and the peripheral leukocytes in the patients with acute cerebral infarction. Methods Sixty-two patients with large artery atherosclerosis cerebral infarction were selected as cerebral infarction group;and 54 age-and gender-matched patients with atherosclerosis and without cerebral cerebrovascular events were recruited control group.The dominant carotid artery plaques were performed by CEUS,the peak of time-intensity curve(TIC-P) and the mean of time-intensity curve (TIC-M) were measured by off-line quantitative analysis.The peak (FC-P), time to peak (FC-TP),sharpness (FC-S) and under the curve area (FC-AUC) were obtained from fitting curves of time-intensity. The correlations between parameters of contrast-enhanced ultrasound and leukocyte counts were analyzed by Pearson correlation analysis. Results ①In the cerebral infarction group, the total leukocytes and neutrophils were higher than those in the control group,while the number of lymphocytes was lower than that of the control group(all P <0.05). ②In the cerebral infarction group,the TIC-P and TIC-M values were greater than those in the control group ( P < 0.05).Compared with the control group,the FC-P,FC-S and FC-AUC values in the cerebral infarction group were increased (all P <0.05). ③There was a negative correlation between PIG-P,TIC-M of FC-P and lymphocytes( r = -0.291,-0.263 and -0.270;all P <0.05).FC-S and FC-AUC were positively correlated with neutrophils ( r =0.261,0.298;all P < 0.05). Conclusions Carotid plaque neovascularizations is related to peripheral leukocyte count.CEUS help us know more the vulnerability of plaque.

Objective To formulate targeted nursing measures and prevent or reduce falls of inpatients through the analysis of related risk factors of falls of spinal degenerative disease patients. Methods Review and analysis falls of inpatients happened between January 2015 and April 2016 in our department and find out that all kinds of dangerous reasons lead to falls. All inpatients risk assessments are evaluated with Morse Fall Scale on admission and during hospitalization. The grading standard of patient safety events was used in the classification of fall outcomes. Results A total of 13 inpatients were all high risk patients by Morse Fall Scale. Risk factors of falls:periods between 0:00 and 2:00, 6:00 and 8:00am, 18:00 and 20:00, in 3 days and 2 weeks after admission; combined diseases such as cardiovascular disease, diabetes, osteoarthritis of the knee; accompanied by muscle strength, muscle tension abnormalities. use related drugs;need of accompany or assisted walking;lack of safety awareness;slippery ground, compressed activity space and lack of light. Conclusions Falls of inpatients with spinal degenerative diseases are caused by the joint action of physiological difference, disease, medicine and weak safety consciousness of inpatients and caregivers, with specialty and occurrence in a certain time. Nursing measures can help to avoid and reduce the falls of inpatients.

Objective To observe the expression dynamics of lens-related transcription factors in human embryonic stem cell (hESC) differentiated into lentoid body(LB).Methods A "three-stage" protocol was used for LB directional differentiation from hESC in vitro.The hESC (D0) and three differentiation stages cells were collected to analyze the expression dynamics of lens development-associated transcription factors by high throughput RNA sequencing technology in hESC-induced LB.Western blot and cell immunofluorescence were used to observe the involved genes at protein level.Results During D0-D6,cells became more round and compact.And during D7-D18,cells morphology gradually changed to spindle.At the end of D35,three-dimensional and transparent structurelentoid body (LB) was obtained.RT-PCR results showed that the stem cell related genes reduced and the lens specific genes increased significantly,and the LB was characterized by the expression of crystallins.According to clustering analysis of high throughput sequencing,a distinct difference in transcription factors gene expression was observed between D0 and D32.Meanwhile,the difference between D6 and D18 was minimum.The expressions of preplacodal genes,including DLX3,DLX5,DLX6,HES1,HES4,OTX2 and EYA1 increased remarkably at the first induction stage and then decreased.Lens-specification gene SOX2 declined gradually and then increased.In addition,the expression of PAX6 increased during all three induction stages.Furthermore,lens-differentiation genes including MAB21L1,CMAF,PROXI and PITX3 had no significant change in the early induction stage,but increased significantly at the third induction stage.Conclusions The expression dynamics of lens development-associated transcription factors in the hESC induced LB corresponded to those in vivo,which indicate that this induction system can recapitulate early lens development well and lay the foundation of studying lens embryonic development andtranscription factor associated congenital lens diseases.

Objective To obtain a thyroid cancer cell line integrated with luciferase and fluorescence,and to establish a metastatic nude mouse model of papillary thyroid cancer monitored with in vivo imaging system. Methods Lentivirus carrying recombinant plasmid containing luciferase gene and mCherry gene infected high malignant papillary thyroid cancer cell line (BHP10-3SCmice). The stable thyroid cancer cell line (BHP10-3mluc) labeled with luciferase and mCherry selected by hygromycin, analyzed for fluorescence with fluorescence microscopy and bioluminescence with in vitro analysis and in vivo imaging. 0.25 ml(5×106/ml)BHP10-3mluc cell suspension were injected into the left thigh intramuscular tissue of 5-week-old nude mice to establish BALB/C nude mice metastatic tumor model. The growth and metastasis of the tumors were monitored weekly with in vivo imaging system. The nude mice were sacrificed on the 42th day and the tissues with metastatic tumors were analyzed by fluorescent imaging ex vivo. The ectopic implanted tumors and metastatic lesions were verified by tissue sections with Hematoxylin and Eosin staining. Results BHP10-3mluc cells showed similar growth characteristics to the original BHP10-3SCmice cells and stably expressed luciferase and mCherry. At the end of the first week after ectopic implantation,the xenograft tumors were found and in the 6th week,the tumors were found to metastasized to adjacent lymph nodes and lungs,which was consistent with the results of pathology.The growth and metastasis of tumors can be accurately monitored with in vivo imaging system. Conclusions A PTC cell line stably expressing bioluciferase and fluorescence was successfully established.The nude mouse model of PTC metastatic tumor generated by intramuscular inoculation of those cells can be well monitored with in vivo imaging system, which provides ideal models for researches on tumor growth, metastasis,and drug treatment.

Objective:To study the association of RAC1 gene polymorphisms with protein expression levels of Rac1-GTP. Methods:A total of 182 healthy Hans population in Hubei were recruited. The 4 tag-SNPs in RAC1 gene were genotyped by Real time TaqMan-MGB genotyping assay. And the Rac1-GTP protein levels in plasma samples from all participants were determined by enzyme linked immunosorbent assays ( ELISA ) . The distribution characteristics of Rac1-GTP expression levels were also analyzed. Furthermore,the expression levels of Rac1-GTP were compared among different genotypes of the 4 tag-SNPs in RAC1 gene. Results:The distribution of Rac1-GTP expression levels was positive skewed in healthy Chinese Hans population. The expression levels were significantly higher in females than in males (P<0. 05),and appeared in decreased trend with age,but without significant differences (P>0. 05). Different expression levels of Rac1-GTP were observed in different genotypes for rs702482 and rs10951982 (P<0. 05). However,no significant difference was found for rs702483 and rs6954996 (P>0. 05). Conclusion:RAC1 genetic polymorphisms can potentially affect the expression levels of Rac1-GTP protein in healthy Chinese Hans population.

Objective:To develop a method for the detection of 10 single nucleotide polymorphisms(SNPs) of Nm23 gene,and to explore the genotypic and allelic distributions of the 10 SNPs in Chinese Hans population in Wuhan.Methods: Two hundreds healthy subjects ,115 men and 85 women included ,were enrolled as DNA sample donors.The real time TaqMan-MGB genotyping assay was used for the determination of the 10 SNPs selected ,and the results were validated by direct gene sequencing.Results:The method established could accurately and quickly screen the genotypes of the 10 SNPs of Nm23/NDPK gene.Distribution frequencies of the 10 SNPs were similar to these in other researches as well as these of HCB.All the loci follow the Hardy-Weinberg equilibrium.Highly linkage disequilibriums were found between rs 16949649 and rs7207370 , rs16949649 and rs34214448 , rs2159359 and rs2302254 , rs2159359 and rs8075231 ,rs2159359 and rs2041296 ,as well as rs2159359 and rs8071647 ,respectively.Four Tag SNPs:rs34214448 , rs2302254 ,rs11868380 and rs2318785 were initially selected by Heploview software.Conclusion:The method established for SNP gen-otyping can meet the needs for rapid analysis of Nm 23 gene polymorphisms ,and may have great values in investigating the association between gene polymorphisms and diseases as well as adverse drug reactions.