Morphological differences in skeletal muscle atrophy of rats with motor nerve and/or sensory nerve injury

Skeletal muscle atrophy occurs after denervation. The present study dissected the rat left ventral root and dorsal root at L4-0 or the sciatic nerve to establish a model of simple motor nerve injury, sensory nerve injury or mixed nerve injury. Results showed that with prolonged denervation time, rats with simple motor nerve injury, sensory nerve injury or mixed nerve injury exhibited abnormal behavior, reduced wet weight of the left gastrocnemius muscle, decreased diameter and cross-sectional area and altered ultrastructure of muscle cells, as well as decreased cross-sectional area and increased gray scale of the gastrocnemius muscle motor end plate. Moreover, at the same time point, the pathological changes were most severe in mixed nerve injury, followed by simple motor nerve injury, and the changes in simple sensory nerve injury were the mildest. These findings indicate that normal skeletal muscle morphology is maintained by intact innervation. Motor nerve injury resulted in larger damage to skeletal muscle and more severe atrophy than sensory nerve injury. Thus, reconstruction of motor nerves should be considered first in the clinical treatment of skeletal muscle atrophy caused by denervation.

SARS-CoV-2 Entry Factors: ACE2 and TMPRSS2 Are Expressed in Peri-Implantation Embryos and the Maternal–Fetal Interface

在全球肆虐的严重急性呼吸综合征冠状病毒2(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2)导致了大量人群感染,血管紧张素转化酶2(angiotensin-converting enzyme 2, ACE2)是SARS-CoV和SARS-CoV-2的受体蛋白。然而是否存在SARS-CoV-2的垂直传播仍颇有争议。为了探讨SARS-CoV-2垂直传播的潜在风险,我们利用已发表的单细胞转录组数据观察了围着床期胚胎和母胎界面上ACE2和TMPRSS2(编码跨膜丝氨酸蛋白酶2)的转录表达情况。结果显示,ACE2和TMPRSS2在第6天(D6)围着床期胚胎的滋养外胚层(trophectoderm, TE)细胞、妊娠第8周的合胞滋养层(syncytiotrophoblast at 8 weeks of gestation, STB8W)细胞以及妊娠第24周的绒毛外滋养层(extravillous trophoblasts at 24 weeks of gestation, EVT24W)细胞中均存在明显的共表达现象,表明这些细胞类群可能易被SARS-CoV-2感染。在这三个细胞类群中,ACE2阳性表达细胞相对于阴性细胞在自噬和免疫相关过程中存在差异。尽管ACE2的表达水平在围着床期胚胎中没有表现出性别偏差,但是其在D6 TE、第6天围着床期胚胎的原始内胚层(D6 primitive endoderm, D6 PE)细胞以及ACE2阳性表达的STB细胞中存在明显的性别差异。这表明围着床期发育第6天以及妊娠期胚胎对SARS-CoV-2的易感性可能存在性别差异。我们的结果揭示了胚胎移植过程中,围着床期以及妊娠期胚胎面临着SARS-CoV-2的潜在感染风险。

Monitoring the in vivo siRNA release from lipid nanoparticles based on the fluorescence resonance energy transfer principle

The siRNA-loaded lipid nanoparticles have attracted much attention due to its significant gene silencing effect and successful marketization.However,the in vivo distribution and release of siRNA still cannot be effectively monitored.In this study,based on the fluorescence resonance energy transfer(FRET)principle,a fluorescence dye Cy5-modified survivin siRNA was conjugated to nanogolds(Au-DR-siRNA),which were then wrapped with lipid nanoparticles(LNPs)for monitoring the release behaviour of siRNA in vivo.The results showed that once Au-DR-siRNA was released from the LNPs and cleaved by the Dicer enzyme to produce free siRNA in cells,the fluorescence of Cy5 would change from quenched state to activated state,showing the location and time of siRNA release.Besides,the LNPs showed a significant antitumor effect by silencing the survivin gene and a CT imaging function superior to iohexol by nanogolds.Therefore,this work provided not only an effective method for monitoring the pharmacokinetic behaviour of LNP-based siRNA,but also a siRNA delivery system for treating and diagnosing tumors.

Study on the Quality of Traditional Chinese Veterinary Medicine and Discussion on the Problems

In this paper,the study on the quality of traditional Chinese veterinary medicine and its existing problems are discussed,which mainly include the research techniques and research thoughts of traditional Chinese veterinary medicine,and the problems in its research progress,at the same time,the causes for the quality problems of traditional Chinese veterinary medicine are analyzed,and the solutions for the problems are put forward. The quality research of traditional Chinese veterinary medicine should refer to traditional Chinese medicine,then inherit,carry forward and innovate it based on the modern research. Solving the problems in the development of traditional Chinese veterinary medicine and putting forward the thinkings conforming to the development of traditional Chinese veterinary medicine using advanced instruments and devices as well as science and technology are urgent affairs.

Inhibition of Sirtuin 2 exerts neuroprotection in aging rats with increased neonatal iron intake

Impaired iron homeostasis may cause damage to dopaminergic neurons and is critically involved in the pathogenesis of Parkinson’s disease. At present, very little is understood about the effect of neonatal iron intake on behavior in aging animals. Therefore, we hypothesized that increased neonatal iron intake would result in signiifcant behavior abnormalities and striatal dopamine depletion during aging, and Sirtuin 2 contributes to the age-related neurotoxicity. In the present study, we observed that neonatal iron intake （120 μg/g per day） during postnatal days 10–17 resulted in significant behavior abnormalities and striatal dopamine depletion in aging rats. Furthermore, after AK-7 （a selective Sirtuin 2 inhibitor） was injected into the substantia nigra at postnatal 540 days and 570 days （5 μg/side per day）, striatal dopamine depletion was signiifcant-ly diminished and behavior abnormality was improved in aging rats with neonatal iron intake. Experimental ifndings suggest that increased neonatal iron intake may result in Parkinson’s dis-ease-like neurochemical and behavioral deifcits with aging, and inhibition of Sirtuin 2 expression may be a neuroprotective measure in Parkinson’s disease.

Study of Cuscuta chinensis Lam. in Nuhuang Fuzheng Oral Liquid by Thin Layer Chromatography

[Objectives] To better control the quality of Nuhuang Fuzheng Oral Liquid and study the main component Cuscuta chinensis Lam.by Thin Layer Chromatography. [Methods] Through changing the treatment methods of the test sample solution,proportion of the developing solvent and sample application volume,taking the spot resolution,definition,and Rf value,optimal Thin Layer Chromatography conditions were screened for Cuscuta chinensis Lam. [Results] After the test sample solution passing the neutral alumina column,it was extracted two times using the ethyl acetate. Methanol was added to dissolve. Benzene-ethyl acetate-formic acid( 5∶5∶2.5) was used as developing solvent.And ammonia fumigation was carried out to develop color. In the thin layer chromatograph obtained through these conditions,Nuhuang Fuzheng Oral Liquid test sample solution showed the same stripe in the same position of the control drug chromatogram and there was no obvious tailing phenomenon and the spot was clear. [Conclusions] The thin layer chromatography identification conditions can be used as the method for quality control of Cuscuta chinensis Lam. in Nuhuang Fuzheng Oral Liquid.

Identification of Citric Acid in Wuhuang Oral Liquid by Thin Layer Chromatography

[Objectives] To screen the identification conditions of citric acid in Wuhuang Oral Liquid by thin layer chromatography,and establish the quality control method for citric acid in Wuhuang Oral Liquid.[Methods] Different treatment methods were adopted for test sample,developing agent,and drying time,thin layer chromatography separation condition and spot definition were taken as indicators to conduct experiment,to select optimal thin layer identification method. [Results] Methanol was used as the extraction solvent,ultrasonic treatment,ether extraction,dissolution by anhydrous ethanol as treatment conditions of test sample; upper solution of butyl acetate-formic acid-water(4 ∶ 2 ∶ 2) after placing one hour was taken as developing agent; 0. 1% bromocresol green(BCG) as the developer; when developing the color in 3 hours after development,in thin layer chromatograph,there appeared the same strip in the same position of test sample of Wuhuang ORAL Liquid and control substance,no obvious trailing phenomenon,and the color was uniform and clear.[Conclusions]The thin layer chromatography identification conditions can be used as the method for quality control of Wuhuang Oral Liquid.

GametesOmics:A Comprehensive Multi-omics Database for Exploring the Gametogenesis in Humans and Mice

Gametogenesis plays an important role in the reproduction and evolution of species.The transcriptomic and epigenetic alterations in this process can influence the reproductive capacity,fertilization,and embryonic development.The rapidly increasing single-cell studies have provided valuable multi-omics resources.However,data from different layers and sequencing platforms have not been uniformed and integrated,which greatly limits their use for exploring the molecular mechanisms that underlie oogenesis and spermatogenesis.Here,we develop GametesOmics,a comprehensive database that integrates the data of gene expression,DNA methylation,and chromatin accessibility during oogenesis and spermatogenesis in humans and mice.GametesOmics provides a user-friendly website and various tools,including Search and Advanced Search for querying the expression and epigenetic modification(s)of each gene;Tools with Differentially expressed gene(DEG)analysis for identifying DEGs,Correlation analysis for demonstrating the genetic and epigenetic changes,Visualization for displaying single-cell clusters and screening marker genes as well as master transcription factors(TFs),and MethylView for studying the genomic distribution of epigenetic modifications.GametesOmics also provides Genome Browser and Ortholog for tracking and comparing gene expression,DNA methylation,and chromatin accessibility between humans and mice.GametesOmics offers a comprehensive resource for biologists and clinicians to decipher the cell fate transition in germ cell development,and can be accessed at http://gametesomics.cn/.

SPEED:an integrated,smartphone-operated,handheld digital PCR Device for point-of-care testing

This study elaborates on the design,fabrication,and data analysis details of SPEED,a recently proposed smartphonebased digital polymerase chain reaction(dPCR)device.The dPCR chips incorporate partition diameters ranging from 50μm to 5μm,and these partitions are organized into six distinct blocks to facilitate image processing.Due to the superior thermal conductivity of Si and its potential for mass production,the dPCR chips were fabricated on a Si substrate.A temperature control system based on a high-power density Peltier element and a preheating/cooling PCR protocol user interface shortening the thermal cycle time.The optical design employs four 470 nm light-emitting diodes as light sources,with filters and mirrors effectively managing the light emitted during PCR.An algorithm is utilized for image processing and illumination nonuniformity correction including conversion to a monochromatic format,partition identification,skew correction,and the generation of an image correction mask.We validated the device using a range of deoxyribonucleic acid targets,demonstrating its potential applicability across multiple fields.Therefore,we provide guidance and verification of the design and testing of the recently proposed SPEED device.

Single-cell transcriptome analysis of the novel coronavirus (SARS-CoV-2) associated gene ACE2 expression in normal and non-obstructive azoospermia (NOA) human male testes

Being infected by SARS-CoV-2 may cause damage to multiple organs in patients, such as the lung, liver and heart. Angiotensin-converting enzyme 2(ACE2), reported as a SARS-CoV-2 receptor, is also expressed in human male testes. This suggests a potential risk in human male reproductive system. However, the characteristics of ACE2-positive cells and the expression of other SARS-CoV-2 process-related genes are still worthy of further investigation. Here, we performed singlecell RNA seq(scRNA-seq) analysis on 853 male embryo primordial germ cells(PGCs) and 2,854 normal testis cells to assess the effects of the SARS-CoV-2 virus on the male reproductive system from embryonic stage to adulthood. We also collected and constructed the scRNA-seq library on 228 Sertoli cells from three non-obstructive azoospermia(NOA) patients to assess the effects at disease state. We found that ACE2 expressing cells existed in almost all testis cell types and Sertoli cells had highest expression level and positive cells ratio. Moreover, ACE2 was also expressed in human male PGCs. In adulthood, the level of ACE2 expression decreased with the increase of age. We also found that ACE2 positive cells had high expressions of stress response and immune activation-related genes. Interestingly, some potential SARS-CoV-2 process-related genes such as TMPRSS2, BSG, CTSL and CTSB had different expression patterns in the same cell type. Furthermore, ACE2 expression level in NOA donors’ Sertoli cells was significantly decreased. Our work would help to assess the risk of SARS-CoV-2 infection in the male reproductive system.

Genetic analysis and preimplantation genetic diagnosis of Chinese Marfan syndrome patients

Marfan syndrome (MFS)(OMIM 154700) is a relatively common autosomal dominant genetic disease that causes skeletal, ocular, and cardiovascular defects and was first described by a French pediatrician in 1896 (Bitterman and Sponseller, 2017). Its prevalence rate is 1/3000—1/5000, and more than 25% of cases are sporadic (Chiu et al., 2014). Studies have shown that about 90% of MFS is caused by variants in the fibrillin-1 gene (FBN1, OMIM 134797). FBN1, located on chromosome 15q21.1, encodes a macromolecular glycoprotein-fibrin 1, which aggregates to form microfibers in the extracellular matrix and distributes in various human connective tissues, such as periosteum, vessel wall, and crystal suspensor ligament. Variants in FNB1 have been reported in 65 exons, but the relationship between genotype and phenotype remains rather unclear (Sakai et al., 2016). Studies have also shown that patients with MFS and similar diseases may have variants in other related genes such as members of the transforming growth factor beta receptor (TGFBR) family (Mizuguchi et al., 2004;Sakai et al., 2006;Bolar et al., 2012;De Cario et al., 2018). For better prevention and treatment of MFS as well as for suspected MFS patients, there is a strong need for efficient genetic testing for early diagnosis and differential diagnoses of patients with related phenotypes (Aubart et al., 2018).

Genetic testing and PGD for unexplained recurrent fetal malformations with MAGEL2 gene mutation

Birth defects are caused by multiple factors,such as chromosome abnormality,environmental factors,and maternal factors.In this study,we focused on exploring the genetic causes of a non-consanguineous couple who suffered from four times of unsuccessful pregnancy due to unexplained recurrent fetal malformations with similar symptoms and normal chromosome copy number variations.Using trio-whole exome sequencing(trio-WES) for this couple and one of the affected fetuses,we found a mutation,c.1996 delC on the maternal imprinted gene MAGEL2 that was carried by the affected fetus and husband,leading to Schaaf-Yang syndrome.To screen this mutation,we further performed preimplantation genetic diagnosis(PGD) strategy followed by a gene pedigree validation and pathogenicity analysis.After the transfer of a PGD-screened embryo,a normal newborn without previous abnormal symptoms was born(February 15,2019).We present the first data that identified a pathogenic gene(MAGEL2 c.1996 delC) in a fetus with Schaaf-Yang syndrome in the EAS(East Asian) database and overcame this genetic defect by using processed PGD for this couple based on the WES results.

Effects of vitrification and cryostorage duration on single-cell RNA-Seq profiling of vitrified-thawed human metaphase Ⅱ oocytes

Oocyte cryopreservation is widely used for clinical and social reasons.Previous studies have demonstrated that conventional slow-freezing cryopreservation procedures,but not storage time,can alter the gene expression profiles of frozen oocytes.Whether vitrification procedures and the related frozen storage durations have any effects on the transcriptomes of human metaphase Ⅱ oocytes remain unknown.Four women(30–32 years old)who had undergone IVF treatment were recruited for this study.RNA-Seq profiles of 3 fresh oocytes and 13 surviving vitrified-thawed oocytes(3,3,4,and 3 oocytes were cryostored for 1,2,3,and 12 months)were analyzed at a single-cell resolution.A total of 1987 genes were differentially expressed in the 13 vitrifiedthawed oocytes.However,no differentially expressed genes were found between any two groups among the 1-,2-,3-,and 12-month storage groups.Further analysis revealed that the aberrant genes in the vitrified oocytes were closely related to oogenesis and development.Our findings indicated that the effects of vitrification on the transcriptomes of mature human oocytes are induced by the procedure itself,suggesting that long-term cryostorage of human oocytes is safe.

Identifying normal embryos from reciprocal translocation carriers by whole chromosome haplotyping

Reciprocal translocation is a chromosomal structural abnormal- ity that arises when two non-homologous chromosomes rearrange and attach with each other, an incidence that occurs in about 1/500 to 1/625 newborns （Mackie and Scriven, 2002）. This event typically does not lead to any significant loss of genetic material, thus recip- rocal translocation carriers do not exhibit any severe abnormal phenotypes （Scriven et al., 1998; Zhang et al., 2016）.

Taurine Transporter dEAAT2 is Required for Auditory Transduction in Drosophila

Drosophila dEAAT2, a member of the excitatory amino-acid transporter(EAAT) family, has been described as mediating the high-affinity transport of taurine, which is a free amino-acid abundant in both insects and mammals. However, the role of taurine and its transporter in hearing is not clear. Here, we report that dEAAT2 is required for the larval startle response to sound stimuli. d EAAT2 was found to be enriched in the distal region of chordotonal neurons where sound transduction occurs. The Ca2+imaging and electrophysiological results showed that disrupted dEAAT2 expression significantly reduced the response of chordotonal neurons to sound.More importantly, expressing d EAAT2 in the chordotonal neurons rescued these mutant phenotypes. Taken together,these findings indicate a critical role for Drosophila dEAAT2 in sound transduction by chordotonal neurons.

Porous β-cyclodextrin nanotubular assemblies enable high-efficiency removal of bisphenol micropollutants from aquatic systems

The performance of water purification by adsorption method has been limited owing to the fact that most of current available adsorbents fail to achieve satisfactory removal performance for organic micropollutants.Herein,we report the design and synthesis of novel porous polymeric adsorbent built fromβ-cyclodextrin(B-CD),in whichβ-CD molecules are arranged in an ordered bis(β-CD)tubular assemblies.The induction of bis(B-CD)units renders them high adsorption affinity toweard bisphenols(bisphenol A and its analogues bisphenol B,bisphenol F and bisphenol S),the typical endocrine disruptors,via the formation of stable host-guest inclusion complexes in aquatic systems.In combination with their high porosity(Brunauer-Emmett-Teller(BET)surface area of 150 m2·g-1),abundantβ-CD content and fast sorption kinetics,the obtained adsorbent outperforms commercial water purifier in elimination of bisphenol micropollutants from potable water.Our work may open a new avenue for designing highly eficient adsorbents for removal of organic micropollutants from aquatic systems.

Aldehyde dehydrogenase ALDH3F1 involvement in flowering time regulation through histone acetylation modulation on FLOWERING LOCUS C

Flowering time regulation is one of the most important processes in the whole life of flowering plants and FLOWERING LOCUS C(FLC)is a central repressor of flowering time.However,whether metabolic acetate level affects flowering time is unknown.Here we report that ALDEHYDE DEHYDROGENASE ALDH3F1 plays essential roles in floral transition via FLC-dependent pathway.In the aldh3f1-1 mutant,the flowering time was significant earlier than Col-0 and the FLC expression level was reduced.ALDH3F1 had aldehyde dehydrogenase activity to affect the acetate level in plants,and the amino acids of E214 and C252 are essential for its catalytic activity.Moreover,aldh3f1 mutation reduced acetate level and the total acetylation on histone H3.The H3K9Ac level on FLC locus was decreased in aldh3f1-1,which reduced FLC expression.Expression of ALDH3F1 could rescue the decreased H3K9Ac level on FLC,FLC expression and also the early-flowering phenotype of aldh3f1-1,however ALDH3F1E214A or ALDH3F1C252A could not.Our findings demonstrate that ALDH3F1 participates in flowering time regulation through modulating the supply of acetate for acetyl-CoA,which functions as histone acetylation donor to modulate H3K9Ac on FLC locus.

Exploring the role of autophagy during early human embryonic development through single-cell transcriptome and methylome analyses

Early human embryogenesis is a very sophisticated process due to its unique gene regulatory network.Autophagy has been suggested to play an important role in mediating the development of early embryonic cells in mammals.However,evidence showing how autophagy regulates early human embryogenesis remains to be further explored.In this study,we systematically investigated the human transcriptome and methylome patterns of autophagy-related(ATG)genes in early embryonic cells at single-cell resolution.We analyzed the transcriptomic data of 365 cells and methylome data of 265 cells.The results showed that most ATG genes remained epigenetically active and were expressed stably throughout early embryogenesis,whereas the dynamics varied among different developmental stages.This evidence indicated that the autophagy pathway was constitutively activated and exerted a fundamental role in early human embryo development.Our work,for the first time,comprehensively reveals the features of autophagy during early human embryo development.

The p.(Pro170Leu) variant in NOG impairs noggin secretion and causes autosomal dominant congenital conductive hearing loss due to stapes ankylosis

Conductive hearing loss is the impairment in the mechanical transduction of sound wave through the external ear and the middle ear.Although most cases are sporadic due to acquired causes such as infections(otitis media and otitis externa),cerumen obstruction,and injuries,congenital structural defects are uncommon for significant etiologies to recognize.Stapes ankylosis is characterized by conductive hearing loss.It may be difficult to differentiate from otosclerosis,the most common cause of progressive conductive hearing loss,by audiologic evaluation,when the diagnosis is delayed.Skeletal anomalies may be subtle,such that the syndrome may not be recognized(Brown et al.,2002).

Axonemal Dynein DNAH5 is Required for Sound Sensation in Drosophila Larvae

Chordotonal neurons are responsible for sound sensation in Drosophila.However,little is known about how they respond to sound with high sensitivity.Using genetic labeling,we found one of the Drosophila axonemal dynein heavy chains,CG9492(DNAH5),was specifically expressed in larval chordotonal neurons and showed a distribution restricted to proximal cilia.While DNAH5 mutation did not affect the cilium morphology or the trafficking of Inactive,a candidate auditory transduction channel,larvae with DNAH5 mutation had reduced startle responses to sound at low and medium intensities.Calcium imaging confirmed that DNAH5 functioned autonomously in chordotonal neurons for larval sound sensation.Furthermore,disrupting DNAH5 resulted in a decrease of spike firing responses to low-level sound in chordotonal neurons.Intriguingly,DNAH5 mutant larvae displayed an altered frequency tuning curve of the auditory organs.All together,our findings support a critical role of DNAH5 in tuning the frequency selectivity and the sound sensitivity of larval auditory neurons.