Improved in situ sequencing for high-resolution targeted spatial transcriptomic analysis in tissue sections.

Spatial transcriptomics enables the study of localization-indexed gene expression activity in tissues, providing the transcriptional landscape that in turn indicates the potential regulatory networks of gene expression. In situ sequencing (ISS) is a targeted spatial transcriptomic technique, based on padlock probe and rolling circle amplification combined with next-generation sequencing chemistry, for highly multiplexed in situ gene expression profiling. Here, we present improved in situ sequencing (IISS) that exploits a new probing and barcoding approach, combined with advanced image analysis pipelines for high-resolution targeted spatial gene expression profiling. We develop an improved combinatorial probe anchor ligation chemistry using a 2-base encoding strategy for barcode interrogation. The new encoding strategy results in higher signal intensity as well as improved specificity for in situ sequencing, while maintaining a streamlined analysis pipeline for targeted spatial transcriptomics. We show that IISS can be applied to both fresh frozen tissue and formalin-fixed paraffin-embedded tissue sections for single-cell level spatial gene expression analysis, based on which the developmental trajectory and cell-cell communication networks can also be constructed.

[Clinical study on effect and therapeutical mechanism of composite Salvia injection on acute cerebral infarction].

OBJECTIVE: To investigate the curative effect and therapeutical mechanism of composite salvia injection (CSI) in treating ischemic cerebral infarction in the respect of oxygen free radical and apolipoprotein. METHODS: Sixty-eight cases of ischemic cerebral infarction within the first 72 hrs after onset were divided randomly into the CSI group (treated with CSI) and the control group (treated with Xueshuantong). Serum lipid peroxide (LPO) and superoxide dismutase (SOD) were measured by colorimetry and apolipoprotein A1 (ApoA1) and ApoB100 were measured with unidirectional immune diffusion method. RESULTS: Serum levels of LPO and ApoB100 were obviously lower, and levels of SOD and ApoA1 significantly higher in the CSI group than those in the control group (P < 0.05 or P < 0.01). The total effective rate of CSI in treating cerebral infarction was 88.24%, which was significantly higher than that of the control (P < 0.05). CONCLUSION: CSI shows definite effect in treating cerebral infarction, to reduce the oxygen free radical damage and regulate the apolipoprotein metabolism possibly was the important therapeutical mechanism.