Development of a High-throughput Sequencing Platform for Detection of Viral Encephalitis Pathogens Based on Amplicon Sequencing.

Objective: Viral encephalitis is an infectious disease severely affecting human health. It is caused by a wide variety of viral pathogens, including herpes viruses, flaviviruses, enteroviruses, and other viruses. The laboratory diagnosis of viral encephalitis is a worldwide challenge. Recently, high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections. Thus, In this study, we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods: We designed nine pairs of specific polymerase chain reaction (PCR) primers for the 12 viruses by reviewing the relevant literature. The detection ability of the primers was verified by software simulation and the detection of known positive samples. Amplicon sequencing was used to validate the samples, and consistency was compared with Sanger sequencing. Results: The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×, and the sequence lengths were consistent with the sizes of the predicted amplicons. The sequences were verified using the National Center for Biotechnology Information BLAST, and all results were consistent with the results of Sanger sequencing. Conclusion: Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis. It is also a useful tool for the high-volume screening of clinical samples.

[Quantitative analysis of craniofacial skeleton asymmetry by three-dimensional computed tomography].

OBJECTIVE: To present a method of quantitative diagnosis of craniofacial skeleton deformities based on three-dimensional computed tomography (3D CT). METHODS: 20 cases with facial asymmetric deformities underwent 3D CT and the 3D images were reconstructed by Mimics 10.0 (Belgium). Anatomical landmarks were located and the coordinate of the landmarks obtained. Axial images of 1 patient with Romberg disease was used as representative case. The differences in the distance between the right landmarks and the left were calculated and analyzed. RESULTS: The measurement results were not significantly different between two stages with an interval of 4 weeks ( P > 0.05), showing a reproducible resutls. The deviation of landmarks at facial midline increased gradually from upward to downward, reaching (2.63 +/- 0.54) mm at menton point. Paired landmarks showed asymmetry in three dimensions, especially gonion point on the left side, which was deviated 10.21 mm inward, 9.26 mm forward, 6.30 mm upward, compared to the opposite side. CONCLUSIONS: The method of 3D CT quantitative analysis can provide precise information in the diagnosis and treatment planning of facial asymmetry deformity.

[Effect on blood pressure and microcirculation of nail fold in primary hypertension patients treated with acupuncture according to syndrome differentiation].

OBJECTIVE: To verify the efficacy of primary hypertension treated with acupuncture at acupoints selected according to syndrome differentiation and probe into the mechanism of acupuncture for primary hypertension. METHODS: One hundred and thirty-five cases of primary hypertension were randomized into an observation group (108 cases) and a control group (27 cases). In either group, Fengchi (GB 20), Quchi (LI 11), Zusanli (ST 36) and Sanyinjiao (SP 6) were selected conventionally. In observation group, on the basis of the acupoints selected above, the supplementary points were selected according to syndrome differentiation in Chinese medicine and the control group was selected main points only, once per day. After 15 days acupuncture, the efficacy and changes in microcirculation of nail fold were observed. RESULTS: The remarkable effective rate and the total effective rate were 29.6% (32/108) and 84.2% (91/108) in observation group respectively, which were superior to 18. 5% (5/27) and 70.4% (19/27) as compared with control group separately (both P < 0.05). After treatment, the microcirculation of nail fold was all improved in two groups, of which blood flow state integral, peripheral capillary loop state integral and the total integral were all improved obviously (all P < 0.05). The improvements in observation group were much more significant than those in control group (all P < 0.05). CONCLUSION: Acupuncture is effective significantly on primary hypertension and the point selection according to syndrome differentiation can improve the efficacy, which is probably relevant with the reduction in the peripheral vascular resistance due to the improvements of microcirculatory state in mechanism.