Loss of SHROOM3 affects neuroepithelial cell shape through regulating cytoskeleton proteins in cynomolgus monkey organoids

Neural tube defects(NTDs)are severe congenital neurodevelopmental disorders arising from incomplete neural tube closure.Although folate supplementation has been shown to mitigate the incidence of NTDs,some cases,often attributable to genetic factors,remain unpreventable.The SHROOM3 gene has been implicated in NTD cases that are unresponsive to folate supplementation;at present,however,the underlying mechanism remains unclear.Neural tube morphogenesis is a complex process involving the folding of the planar epithelium of the neural plate.To determine the role of SHROOM3 in early developmental morphogenesis,we established a neuroepithelial organoid culture system derived from cynomolgus monkeys to closely mimic the in vivo neural plate phase.Loss of SHROOM3 resulted in shorter neuroepithelial cells and smaller nuclei.These morphological changes were attributed to the insufficient recruitment of cytoskeletal proteins,namely fibrous actin(F-actin),myosin II,and phospho-myosin light chain(PMLC),to the apical side of the neuroepithelial cells.Notably,these defects were not rescued by folate supplementation.RNA sequencing revealed that differentially expressed genes were enriched in biological processes associated with cellular and organ morphogenesis.In summary,we established an authentic in vitro system to study NTDs and identified a novel mechanism for NTDs that are unresponsive to folate supplementation.

Albuterol in the treatment of acute respiratory distress syndrome: A meta-analysis of randomized controlled trials

BACKGROUND: This meta-analysis of randomized controlled trials aimed to systematically evaluate the value of albuterol in the treatment of patients with acute respiratory distress syndrome(ARDS).DATA SOURCES: Randomized controlled trials on albuterol treatment of ARDS from its inception to October 2014 were searched systematically. The databases searched included: Pub Med, Ovid EMBASE, Ovid Cochrane, CNKI, WANFANG and VIP. The trials were screened according to the pre-designed inclusion and exclusion criteria. We performed a systematic review and meta-analysis of the randomized controlled trials(RCTs) on albuterol treatment, attempting to improve outcomes, i.e. lowering the 28-day mortality and ventilator-free days.RESULTS: Three RCTs involving 646 patients met the inclusion criteria. There was no significant decrease in the 28-day mortality(risk difference=0.09; P=0.07, P for heterogeneity=0.22, I2=33%). The ventilator-free days and organ failure-free days were significantly lower in the patients who received albuterol(mean difference=–2.20; P<0.001, P for heterogeneity=0.49, I2=0% and mean difference=–1.71, P<0.001, P for heterogeneity=0.60, I2=0%).CONCLUSIONS: Current evidences indicate that treatment with albuterol in the early course of ARDS was not effective in increasing the survival, but significantly decreasing the ventilator-free days and organ failure-free days. Owing to the limited number of included trails, strong recommendations cannot be made.

Diverse coagulopathies in a rabbit model with different abdominal injuries

BACKGROUND: Although coagulopathy can be very common in severe traumatic shock patients, the exact incidence and mechanism remain unclear. In this study, a traumatic shock rabbit model with special abdomen injuries was developed and evaluated by examining indicators of clotting and fi brinolysis.METHODS: Forty New Zealand white rabbits were randomly divided into four groups: group 1(sham), group 2(hemorrhage), group 3(hemorrhage-liver injury), and group 4(hemorrhage-liver injury/intestinal injury-peritonitis). Coagulation was detected by thromboelastography before trauma(T0), at 1 hour(T1) and 4 hours(T2) after trauma.RESULTS: Rabbits that suffered from hemorrhage alone did not differ in coagulation capacity compared with the sham group. The clot initiations(R times) of group 3 at T1 and T2 were both shorter than those of groups 1, 2, and 4(P<0.05). In group 4, clot strength was decreased at T1 and T2 compared with those in groups 1, 2, and 3(P<0.05), whereas the R time and clot polymerization were increased at T2(P<0.05). The clotting angle signifi cantly decreased in group 4 compared with groups 2 and 3 at T2(P<0.05).CONCLUSION: This study suggests that different abdominal traumatic shock show diverse coagulopathy in the early phase. Isolated hemorrhagic shock shows no obvious effect on coagulation. In contrast, blunt hepatic injury with hemorrhage shows hypercoagulability, whereas blunt hepatic injury with hemorrhage coupled with peritonitis caused by a ruptured intestine shows a tendency toward hypocoagulability.

非人灵长类体内广泛存在AAV和Cas蛋白的体液免疫应答

新兴的CRISPR/Cas系统因其高效性和便捷性已成为目前最常用的基因编辑工具,并有望在体内基因治疗研究中发挥重要作用.AAV递送系统和CRISPR编辑系统是开展体内基因治疗的重要元件,然而,有研究发现人体中具有针对AAV和Cas9蛋白的预存抗体,说明免疫反应是基因修复治疗必须面对的问题.本研究检测到猕猴(恒河猴和食蟹猴)体内普遍存在针对AAV(包括AAV2、AAV8和AAV9)和Cas蛋白(包括SaCas9、SpCas9、AsCas12a和LbCas12a)的预存抗体.当给小鼠和猕猴体内注射了AAV9或SpCas9蛋白后,均激发了一系列的免疫反应.此外,猕猴体内AAV携带Cas9基因的治疗试验也显示体液免疫反应被激活.上述研究结果进一步证实猕猴与人类似,体内普遍存在AAV和Cas蛋白的预存抗体,且AAV-CRISPR体内基因治疗会引发相应的免疫应答.

Promising therapeutic approaches of utrophin replacing dystrophin in the treatment of Duchenne muscular dystrophy

Duchenne muscular dystrophy(DMD)is a serious genetic neuromuscular rare disease that is prevalent and caused by the mutation/deletion of the X-linked DMD gene that encodes dystrophin.Utrophin is a dystrophin homologous protein on human chromosome 6.Dystrophin and utrophin are highly homologous.They can recruit many dystrophin-glycoprotein complex(DGC)-related proteins and co-localize at the sarcolemma in the early stage of human embryonic development.Moreover,utrophin is overexpressed naturally at the mature myofiber sarcolemma in DMD patients.Therefore,utrophin is considered the most promising homologous protein to replace dystrophin.This review summarizes various modulating drugs and gene therapy approaches for utrophin replacement.As a universal method to treat DMD disease,utrophin has a promising therapeutic prospect and deserves further investigation.