Zebrafish as an animal model for the antiviral RNA interference pathway.

The zebrafish (Danio rerio) possesses evolutionarily conserved innate and adaptive immunity as a mammal and has recently become a popular vertebrate model to exploit infection and immunity. Antiviral RNA interference (RNAi) has been illuminated in various model organisms, including Arabidopsis thaliana, Drosophila melanogaster, Caenorhabditis elegans and mice. However, to date, there is no report on the antiviral RNAi pathway of zebrafish. Here, we have evaluated the possible use of zebrafish to study antiviral RNAi with Sindbis virus (SINV), vesicular stomatitis virus (VSV) and Nodamura virus (NoV). We find that SINVs and NoVs induce the production of virus-derived small interfering RNAs (vsiRNAs), the hallmark of antiviral RNAi, with a preference for a length of 22 nucleotides, after infection of larval zebrafish. Meanwhile, the suppressor of RNAi (VSR) protein, NoV B2, may affect the accumulation of the NoV in zebrafish. Furthermore, taking advantage of the fact that zebrafish argonaute-2 (Ago2) protein is naturally deficient in cleavage compared with that of mammals, we provide evidence that the slicing activity of human Ago2 can virtually inhibit the accumulation of RNA virus after being ectopically expressed in larval zebrafish. Thus, zebrafish may be a unique model organism to study the antiviral RNAi pathway.

Relationship between dental calcification of mandibular teeth and cervical vertebrae maturity in patients with unilateral complete cleft lip and palate.

OBJECTIVE: The purpose of this study was to investigate the relationship between the dental calcification stage (DCS) of the mandibular teeth and the cervical vertebral maturation stage (CVMS) in patients with unilateral complete cleft lip and palate (UCLP). METHODS: One hundred sixty-two UCLP patients (100 males and 62 females) between 8-16 years old were included in this study. The DCS was estimated by the Demirjian method and was converted to the dental age (DA). The CVMS was evaluated by the Baccetti method. The DA of mandibular teeth on two sides of the cleft were analyzed using a t-test. Spearman correlation was used to study the association between CVMS and DCS. The correlation coefficient between the two sides of the cleft was then compared. RESULTS: The total DA was significantly smaller on the cleft side than on the noncleft side in males (p = 0.022). The Spearman rank correlation coefficient revealed a significant correlation between the DCS of each examined tooth and the CVMS (r = 0.627-0.793 in males and r = 0.806-0.899 in females). Additionally, the correlation of the two sides was not significantly different (p > 0.05). The DCS of the first premolar showed the strongest correlation with the CVMS. CONCLUSION: The results confirm the utility of the DCS on both sides of the mandible in male UCLP patients as a simple first-level diagnostic test to evaluate growth and development. The findings also indicate that both the DCS and the CVMS should be assessed if the maturity stage of a growing UCLP patient is relevant to clinical practice.

Altering Intracellular Localization of the RNA Interference Factors by Influenza A Virus Non-structural Protein 1.

Influenza A virus (IAV) causes seasonal infections and periodic pandemics in humans. The non-structural protein 1 (NS1) of IAV is the main viral antagonist of the innate immune responses that play a key role in influenza pathogenesis. However, the mechanism to disrupt the host cell homeostasis by IAV NS1 remains poorly understood. Here, we show that expression of NS1 from the WSN strain, but not PR8 strain, of IAV, markedly induced nuclear import of the host RNA interference (RNAi) factors such as Argonaute-2 and microRNA 16. We found that the single residue substitution of aspartic acid with histidine at position 101 (D101H) of IAV-PR8 NS1 was sufficient to induce the nuclear import process and to enhance the virulence of IAV-PR8 in mice. However, we observed no significant differences between the wild-type and mutant IAV-PR8 in virus titers or induction of the interferon response in lung tissues, indicating a novel role of NS1 in the virulence determination of IAV in a mammalian host. Moreover, our bioinformatic analysis of 69,057 NS1 sequences from all IAV subtypes deposited in the NCBI database revealed that the NS1-H101 gene of IAV-WSN was widespread among H1N1 viruses isolated in 1933 but disappeared completely after 1940. Thus, IAV NS1 (H101) is a mutation selected against during evolution of IAV, suggesting that mutation H101 confers an important biological phenotype.

[Clinical observation of acupuncture at Qianlianquan (Extra) and eight confluent points on chronic pharyngitis].

OBJECTIVE: To observe the clinical therapeutic effect of acupuncture at Qianlianquan (Extra) and eight confluent points on chronic pharyngitis. METHODS: A total of 42 patients with chronic pharyngitis were randomized into an observation group and a control group, 21 cases in each one. In the observation group, acupuncture was applied at Qianlianquan (Extra) and eight confluent points [Neiguan (PC 6), Waiguan (TE 5), Lieque (LU 7), Houxi (SI 3), Gongsun (SP 4), Shenmai (BL 62), Zhaohai (KI 6) and Zulinqi (GB 41)], once a day, 6 days as one course, with an interval of 1 day between the courses, 4-course treatment was required. In the control group, Qinghou Liyan granules were taken orally 5 g each time, 3 times a day, 7 days as one course and 4 courses were required. Before and after treatment, symptoms and signs scores of chronic pharyngitis and the contents of IL-2 and TNF-a in serum were compared in the two groups. RESULTS: Compared before treatment, symptoms and signs scores of chronic pharyngitis and the contents of IL-2 and TNF-a in serum were improved in both groups (all P<0.05), and the changes in the observation group were superior to the control group (all P<0.05). The total effective rate was 90.5% (19/21) in the observation group, which was superior to 81.0% (17/21) in the control group (P<0.05). CONCLUSION: The therapeutic effect of acupuncture at Qianlianquan (Extra) and eight confluent points on chronic pharyngitis is superior to conventional Chinese patent medication treatment by Qing Liyan granules.

[The symmetry of nasal maxillary complex of patients with unilateral complete cleft lip and palate at the stage of mixed dentition-A cone beam CT study].

PURPOSE: To investigate the symmetry of nasal maxillary complex in the three-dimensional directions of patients with unilateral complete cleft lip and palate(UCLP) at the stage of mixed dentition. METHODS: Craniofacial cone-beam CT(CBCT) images of 20 UCLP patients at the stage of mixed dentition were selected. Three-dimensional reconstruction of bone tissue was carried out by Mimics software, nasal root point and 32 markers with the same name of the healthy side and the cleft side of the nasal maxillary complex were traced, three-dimensional reference planes were set up, then the distances between the points to the three-dimensional planes were measured respectively. SPSS 22.0 software package was used to analyze the data. RESULTS: In the horizontal direction, compared with the non-cleft side, the width of the cleft side of the maxilla at INM and SPC was smaller, the width of maxilla at LPA and SPr was larger, the differences were statistically significant (P<0.05). In the sagittal direction, compared with the non-cleft side, the depth of LPA, Maz and SPr at the cleft side was larger, the depth of SPM and SPC was smaller, the differences were statistically significant (P<0.05).In the vertical direction, the positions of LPA, INM and Maz at the cleft side was lower than the non-cleft side, SPC and SPr were higher, the differences were statistically significant(P<0.05). The remaining points had no significant difference (P>0.05). CONCLUSIONS: The nasal maxillary complex of UCLP patients at the stage of mixed dentition is asymmetric in three directions. The asymmetric locations were mainly located in the nasal cavity and alveolar bone. No obvious asymmetry is found in the orbital region and the maxillary region far from the cleft.

[Association between cervical vertebral maturation stages and dental calcification stages in patients with unilateral complete cleft lip and palate].

OBJECTIVE: The purpose of the study is to investigate the relationship between dental calcification stages (DCS) and cervical vertebral maturation stages (CVMS) in patients with unilateral complete cleft lips and palates (UCLP) and to provide a theoretical basis for the treatment time selection of cleft lip and palate (CLP) patients. METHODS: A total of 123 UCLP patients and 215 non-CLP subjects were selected. The DCS of the left mandibular canine, premolar, and second molar in non-CLP subjects and on both cleft sides of UCLP patients were assessed utilizing the Demirjian method. CVMS was observed utilizing the Baccetti method. The results were analyzed by Spearman rank correlation, and the correlation coefficients were compared. RESULTS: There was a correlation between the CVMS and the DCS of the left mandibular canine, the first premolar, the second premolar, and the second molar in the non-CLP subjects and on both cleft sides of the UCLP patients (r=0.762-0.864, P<0.05; r=0.809-0.914, P<0.05, respectively). The correlation between the CVMS and the DCS of the mandibular first premolar was highest among the UCLP patients. Except for the first and the second premolars of UCLP females, the correlation between the DCS and the CVMS of the other teeth did not differ among the non-CLP subjects (P>0.05). CONCLUSIONS: DCS can be utilized as a biological index to determine the growth development statuses. The correlation between the CVMS and the DCS of the mandibular first premolar was the highest.