GABARAP, a microtuble-associated protein, is identified to interact with GABAA receptor. Anchoring of the GABAA receptor to GABARAP helps to cluster the receptor at the synaptic termini and to mediate fast synaptic transmission. GABARAP may mediate interaction of gephyrin with the GABAA receptor to stabilize clusters by forming multimeric structures. Furthermore, GABARAP and gephyrin may play more of a role in receptor sorting and transport to the cell surface than in anchoring to the cytoplasm, because at inhibitory synapses GABARAP appears to associate with transport vesicles rather than the cell surface. The association of GABARAP with NSF (N-ethylmaleimide sensitive factor), a protein involved in intracellular vesicle transport, supports this hypothesis. We cloned cDNA encoding full-length human GABARAP by nested PCR and inserted it into eukaryon expression vector pcDNA6HA and GST fusion protein expression vector pGEX4T2. The recombinant plasmid pGEX4T2-hGABARAP was transformed into E. coli BL21, from which GST-hGABARAP fusion protein was purified after IPTG induction by affinity chromatography with glutathione Sepharose-4B column. The antiserum against GABARAP was generated by immunizing rabbits with the purified GST-hGABARAP and was purified with GST-hGABARAP coupled NHS-activated Sepherose 4 column. The purified polyclonal antibody was effective for Western blotting and immunostaining. The hGABARAP was located both in the cytoplasm and nucleus with an abundant distribution around the peripheral nucleus.
Adaptor Proteins, Signal Transducing ; biosynthesis ; genetics ; immunology ; Animals ; Antibodies ; blood ; immunology ; Antibodies, Monoclonal ; biosynthesis ; genetics ; immunology ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Glutathione Transferase ; biosynthesis ; genetics ; Humans ; Immunization ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; immunology ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

Sepsis and septic shock are important clinical problems in critically ill patients, accounting for the first cause of death in intensive care unit (ICU). Therefore, early diagnosis and treatment are particularly important. Recently, genome-wide expression analysis of non-coding RNA in septic patients showed that more than 80% were differentially expressed compared with healthy individuals. These molecules play important roles in biological processes, including innate immunity, mitochondrial function and apoptosis. Therefore, a class of non-coding RNAs such as microRNA (miRNA), long-chain non-coding RNA (lncRNA) and circular non-coding RNA (circRNA) are increasingly recognized as a regulator of various signaling pathways. The potential of regulatory non-coding RNA target to treat sepsis was discussed by studying non-coding RNAs that might serve as molecular markers of sepsis, and its clinical value was evaluated.

Objective: To explore the related factors of myopia among children and adolescents in Yunnan Province, and to predict and evaluate the influencing factors, so as to provide a scientific theoretical basis for the prevention and control of myopia. Methods: From March 9 to 14, 2023, 848 students from 6 primary and secondary schools in Dali and Lijiang of Yunnan Province were selected by multi stage stratified random cluster sampling method for visual acuity detection and questionnaire survey on myopia related factors. Multivariate Logistic regression analysis was used to establish a Nomogram prediction model for the selected influencing factors. Results: The overall myopia rate of the respondents was 68.3%, the myopia rate of boys (63.4%) was lower than that of girls (72.9%), and the myopia rate of primary school students (46.7%) was lower than that of junior high school students (81.1%), and the difference was statistically significant（ χ 2=8.71, 108.07, P <0.05). Daily eye exercises, activities outside the teaching building during recess, having daily sleep time of 7-9 and >9 h, having both parents without myopia were negatively correlated with the occurrence of myopia in children and adolescents in Yunnan Province ( OR=0.64, 0.63, 0.56, 0.28, 0.48, P < 0.05 ). The reading and writing time after school ≥3 h per day and parents unrestricted time to play video games were positively correlated with myopia ( OR=1.94, 1.78, P <0.05). Based on the influencing factors, a Nomogram prediction model was established to quantitatively evaluate the risk of myopia. The results showed that greater risk for myopia was associated with sleep duration, parental history of myopia, and the time spent reading and writing after school every day. Conclusion Both genetic factors and environmental factors are related to myopia in children and adolescents. The prediction model of nomogram is beneficial for screening high risk factors of myopia and taking corresponding prevention and treatment measures.