Activity

Finally, we validate how CRISpy-pop can accurately predict the activity of guide RNAs across strains using population genomic data.In the developing Xenopus tadpole visual system, the targeting and branching of optic axons in the brain is a dynamic process that is closely intertwined with the morphological differentiation and maturation of their postsynaptic neurons and with the formation, stabilization, and elimination of functional synapses. The coordinated addition and retraction of axonal and dendritic branches guides the gradual recognition between pre- and postsynaptic neuronal partners, which subsequently allows synaptic connections to be formed. Axon and dendrite branching and selective synapse formation and stabilization are developmental mechanisms largely orchestrated by an array of signaling molecules that interact in vivo for the proper formation of functional visual circuits. In vivo real-time imaging of individual fluorophore-labeled neurons in living Xenopus tadpoles has allowed investigation of molecular and cellular mechanisms mediating circuit assembly at a cellular level in the intact organism. In this protocol, we describe the use of bulk and single-cell electroporation to rapidly and efficiently transfect individual retinal ganglion cells (RGCs) with different reagents and to simultaneously visualize optic axon arbor morphology and presynaptic sites in real time. Similar techniques for labeling and visualizing RGC axons can be combined with the use of morpholino antisense oligonucleotides, as we describe here, to alter gene expression cell autonomously.

Gap junctions play a pivotal role in contributing to the formation of astroglial networks and in chronic pain. However, the mechanisms underlying the dysfunction of astroglial gap junctions in chronic pain have not been fully elucidated.

Chronic constriction injury (CCI) of the sciatic nerve was used to establish rat neuropathic pain model. C6 cells were used to perform experiments in vitro. Von Frey hairs and Hargreave’s method were used to determine the withdrawal threshold of rats. Protein expression was detected by immunofluorescence and western blotting.

Astragaloside IV (AST IV) significantly attenuated neuropathic pain and suppressed the excitation of spinal astrocytes in rats with CCI. The antinociceptive effect of AST IV was reversed by the gap junction decoupler carbenoxolone (CBX). STZ inhibitor mouse AST IV inhibited the high expression of phosphorylated connexin 43 (p-Cx43) and p-c-Jun N-terminal kinase (p-JNK) in spinal cord of rats with CCI. JNK inhibitor alleviated neuropathic pain, which was reversed by CBX. JNK inhibitor decreased the high expression of p-Cx43 in both rats with CCI and tumor necrosis factor-alpha (TNF-α)-treated C6 cells. Additionally, the analgesic effect of AST IV was reversed by the adenosine triphosphate-sensitive potassium (K

) channel blocker, glibenclamide (Glib). Glib abolished the inhibitory effects of AST IV on p-JNK and p-Cx43 both in vivo and in vitro. K

channel opener (KCO) mimicked the inhibitory effects of AST IV on p-JNK and p-Cx43 in TNF-α-treated C6 cells.

Our results indicate that the sciatic nerve CCI induces the dysfunction of gap junctions in the spinal cord by activating K

/JNK signaling to contribute to neuropathic pain. AST IV attenuates neuropathic pain via regulating the K

-JNK gap junction axis.

Our results indicate that the sciatic nerve CCI induces the dysfunction of gap junctions in the spinal cord by activating KATP/JNK signaling to contribute to neuropathic pain. AST IV attenuates neuropathic pain via regulating the KATP-JNK gap junction axis.

To explore the relationships between gastric cancer and serum pepsinogen I (PG I), PG II, PG I/II ratio, gastrin 17 (G-17) and

infection, and to investigate dietary and lifestyle risk factors for gastric cancer in Fujian Province, China.

A hospital-based, 11 matched case-control study.

Patients with newly diagnosed gastric cancer were recruited from the Fujian Provincial Hospital and the No. 900 Hospital of the Joint Support Force of the Chinese People’s Liberation Army between July 2014 and December 2016.

A total of 180 pairs of patients with gastric cancer and control subjects were recruited in the study, including 134 (74.4%) male pairs and 46 (25.6%) female pairs.

Serological indicators were tested with ELISA kits. Dietary, lifestyle and psychological factors were investigated through face-to-face questionnaire. Relationships between gastric cancer and these influencing factors were examined by Χ

test and conditional logistic regression.

Serum PG II and G-17 levels and

infection rate w diagnosis of gastric cancer. Increase in serum G-17 level might indicate the location of gastric cancer. Increase in serum PG II level and decrease in PG I/II ratio might imply the clinical stage. Eating hot food, eating pickled vegetables and often feel troubled may be risk factors for gastric cancer, while eating fresh fruits, eating onion or garlic, and drinking tea may be protective factors against the disease.

Study results showed that serum PG, G-17 and H. pylori antibodies could be useful indicators for early diagnosis of gastric cancer. Increase in serum G-17 level might indicate the location of gastric cancer. Increase in serum PG II level and decrease in PG I/II ratio might imply the clinical stage. Eating hot food, eating pickled vegetables and often feel troubled may be risk factors for gastric cancer, while eating fresh fruits, eating onion or garlic, and drinking tea may be protective factors against the disease.

The ageing population and prevalence of long-term disorders with multimorbidity are a major health challenge worldwide. The associations between comorbid conditions and mortality risk are well established; however, few prospective community-based studies have reported on prior risk factors for incident hospital admissions with multimorbidity. We aimed to explore the independent associations for a range of demographic, lifestyle and physiological determinants and the likelihood of subsequent hospital incident multimorbidity.

We examined incident hospital admissions with multimorbidity in 25 014 men and women aged 40-79 in a British prospective population-based study recruited in 1993-1997 and followed up until 2019. The determinants of incident multimorbidity, defined as Charlson Comorbidity Index ≥3, were investigated using multivariable logistic regression models for the 10-year period 1999-2009 and repeated with independent measurements in a second 10-year period 2009-2019.

Between 1999 and 2009, 18 179 participants (73% of the population) had a hospital admission.