A Novel Retrograde AAV Variant for Functional Manipulation of Cortical Projection Neurons in Mice and Monkeys

Retrograde adeno-associated viruses(AAVs)are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks.However,few retro-grade AAV capsids have been shown to offer access to cor-tical projection neurons across different species and enable the manipulation of neural function in non-human primates(NHPs).Here,we report the development of a novel retro-grade AAV capsid,AAV-DJ8R,which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques.In addition,intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations.Moreover,AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen.These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations.

Effects of Dietary Protein and Fat Levels on Growth Performance and Serum Biochemical Parameters of Male Mink

This experiment was conducted to evaluate the effects of dietary protein and fat levels on growth performance and serum biochemical parameters of male minks during the growing period. A total, of 90 healthy male minks at the age of 60 d were randomly assigned into six groups with 15 replicates per group and one mink per replicate. The minks were fed six experimental diets with two protein levels （32% and 36% ） and three fat levels （ 10% , 20% and 30% ） for a 7-day preliminary period and then for 60-day experimental period. The results showed that the final body weight and average daily weight gain （ADWG） were significantly higher in low-protein, middle-fat group and low-protein, high-fat group than those in low-protein, low-fat group and high-protein, low-fat group （P 〈 0. O1 ）. The final weight and ADWG were increased with dietary fat level increasing, and the two indices of 20% and 30% fat groups were significantly higher than those of 10% fat groups （ P 〈 0. O1 ）. The ratio of feed intake to weight gain （F/G） in low-protein, high-fat group was the lowest, which was significantly lower than that in low- protein, low-fat group and high-protein, high-fat group （P 〈 0.05 ）. Serum urea nitrogen （UN） in low-protein, high-fat group was significantly lower than that in other groups （P 〈 0. 01 ）. Serum alanine transaminase （ALT） activity in 32% protein groups was significantly higher than that in 36% protein groups （P 〈 O. 05）. Serum triglyceride （TG） content in low-protein, high-fat group and high-protein, high-fat group was significantly higher than that in low-protein, low-fat group and high-protein, low-fat groups （P 〈 0.05 ）. Serum TG, total cholesterol （TCHO） and high-density lipoprotein （HDL） contents increased with the increase in dietary fat level, and they were all the highest in low-protein, high-fat group. The growth performance and serum biochemical parameters of minks were not affected by the interaction between dietary protein and fat levels （P 〉 O. 05 ）. Considering all the factors, when the dietary protein is 32% and the fat level is 20% to 30% under the condition of this experiment, the dietary protein and fat utilization rates are higher, and the male minks during the growing period can get a better growth performance.

Body Size but Not Food Size Determined Head Sexual Dimorphism in Rana kukunoris from the Tibetan Plateau

Sexual dimorphism between males and females is a common pattern observed in the natural world and has become one of the main focuses of evolutionary biology.Amphibian head size and shape are important factors that may influence food acquisition,but knowledge about the sexual differences between amphibian head size and shape is relatively scant.Here,intersexual differences in the head size and shape of Rana kukunoris,which is a high-elevation frog from the Qinghai-Tibet Plateau,were investigated.Significant differences were found in the head shape but not the head size between sexes.A significant correlation was also observed between prey size and head size.However,these correlations disappeared when individual body size was controlled,suggesting that body size but not food size was the main factor resulting in intersexual differences in head size/shape.The results suggested that food competition between sexes did not result in sexual dimorphism of head shape among these frogs.

Characterization and Phylogenetic Implications of the Complete Mitogenomes of Two Species in the Genus Zhangixalus(Anura:Rhacophoridae)

Mitochondrial genomes(mitogenomes)in frogs are essential for reconstructing the phylogenetic relationships and shedding light on the molecular evolution in these animals.However,there are only seven complete Rhacophoridae genomes that have been reported to date.In this paper,two complete mitogenomes of the Chinese whipping frog(Zhangixalus chenfui)and Emei tree frog(Z.omeimontis)were described,each of which was 20520 and 19782 bp in size,and had A+T contents of 64.26%and 63.83%.The two mitogenomes each included two non-coding control region(D-loop,CR),two ribosomal RNA genes(rRNAs),13 protein-coding genes(PCGs),and 22 transfer RNA genes(tRNAs),and it was found that the mitogenome of Z.chenfui also includes three tandem tRNAMet gene sequences.A typical clover-leaf structure was found for all tRNAs except for tRNASer1(AGN),which showed a reduced DHU arm.The putative D-loop region contains multiple types of tandem repeats regions.Both of these two mitogenomes showed similar pattern of gene rearrangement(tRNA-Ser-ND6-tRNA-Glu-CytbCR1-ND5-CR2-tRNA-Thr-tRNA-Leu-tRNA-Pro).Additionally,three consecutive tRNAMet genes were found for the first time in Z.chenfui,a species in the Rhacophoridae family.For all phylogenetic analyses,which were based on 13 protein-coding genes from 91 Ranoidea mitogenomes,the same phylogenetic trees were observed using either maximum likelihood or Bayesian approaches.These results suggest that the phylogenetic hypotheses for the Ranoidea(including Rhacophoridae,Mantellidae,Ranidae and Dicroglossidae)derived from these mitogenomic data could provide substantiation for the relationships of(Dicroglossidae(Ranidae,(Mantellidae,Rhacophoridae))),and support the presence of a monophyletic group in four families.Moreover,Z.omeimontis and Z.dennysi were found to cluster on the same branch,indicating that they were more closely related as a group.This group may in turn form sister groups with Z.arboreus and Z.schlegelii.However,Z.chenfui is located on the base of other species in the genera.Increased mitogenome sampling should be conducted to provide a more satisfactory resolution to the phylogeny of the Rhacophorus,Rhacophoridae,and Mantellidae.

MicroRNA Expression Signature in Gastric Cancer

Objective： To identify the miRNA specific signature as novel diagnostic and prognostic tools for gastric cancer. Methods： miRNAs expression profiling of 3 normal gastric tissues, 24 malignant tissues, gastric cancer cell SGC7901 and normal gastric cell GES-1 were detected using microarray technology. The hierarchical clustering algorithm of the Cluster software was used to analyse the miRNAs expression of all samples. The expression levels of miR-433 and miR-9 which were significantly down-regulated in gastric cancer tissues and SGC7901 cells by microarray technology were validated by quantitative Real-time PCR （qRT-PCR）. Results： Differential expressions of 26 individual miRNAs between normal samples （including 3 normal gastric tissues and GES-1 cells） and carcinomas （including 24 malignant tissues and SGC7901 cells） were discovered, 19 of them showing down-regulation and 7 up-regulation in carcinoma samples. Hierarchical clustering of the Cancer samples by their miRNA expression accurately separated the carcinomas from normal samples and further their histotypes of carcinomas. The expression levels of miR-433 and miR-9 were significantly down-regulated in gastric cancer tissues and SGC7901cells Conclusion： The differential expression of miR-433 and miR-9 may be used as a novel diagnostic tool for gastric cancer.

Detection of Multi-drug Resistant Acinetobacter Lwoffii Isolated from Soil of Mink Farm

There were 4 Acinetobacter lwoffii obtained from soil samples.The antimicrobial susceptibility of the strains to 16 antimicrobial agents was investigated using K-B method.Three isolates showed the multi-drug resistance.The presence of resistance genes and integrons was determined using PCR.The aadA 1,aac（3＇）-IIc,aph（3＇）-VII,aac（6＇）-Ib,sul2,cat2,floR,and tet（K）genes were detected,respectively.

Quantitative proteomics reveals key pathways in the symbiotic interface and the likely extracellular property of soybean symbiosome

An effective symbiosis between legumes and rhizobia relies largely on diverse proteins at the plantrhizobium interface for material transportation and signal transduction during symbiotic nitrogen fixation.Here,we report a comprehensive proteome atlas of the soybean symbiosome membrane(SM),peribacteroid space(PBS),and root microsomal fraction(RMF)using state-of-the-art label-free quantitative proteomic technology.In total,1759 soybean proteins with diverse functions are detected in the SM,and 1476 soybean proteins and 369 rhizobial proteins are detected in the PBS.The diversity of SM proteins detected suggests multiple origins of the SM.Quantitative comparative analysis highlights amino acid metabolism and nutrient uptake in the SM,indicative of the key pathways in nitrogen assimilation.The detection of soybean secretory proteins in the PBS and receptor-like kinases in the SM provides evidence for the likely extracellular property of the symbiosome and the potential signaling communication between both symbionts at the symbiotic interface.Our proteomic data provide clues for how some of the sophisticated regulation between soybean and rhizobium at the symbiotic interface is achieved,and suggest approaches for symbiosis engineering.

Regulations of Antler Generation and Regeneration by Nutritional Factors

Antler, one of the secondary sex characteristics of male deer, is a highly valued traditional Chinese medicine. The generation and regeneration of deer antler is regulated not only by hormones, but also by nutritional factors. Nutritional factors have great influences on pedicle generation, first antler generation and antler development. Therefore, this paper summarizes recent advances in the study of the influences of nutritional factors on antler generation and regeneration, analyzes the pathways regulating antler generation and regeneration by nutritional factors, and proposes thai nutritional factors regulale antler regeneralion by direct pathway and indirect pathway.

Safety of a Live Attenuated Infectious Bovine Rhinotracheitis Vaccine IBRV LNM Strain

The paper was to evaluate the vaccine safety,and to prevent public health risk due to virus spread,the approach vaccination of was adopted in this research; and neck intramuscular injection of IBRV LNM attenuated vaccine strain was carried out. Blind passage for three generations in animal has tested the reversion risk to virulence. A total of 14 healthy and weaning cows at 6- 8 month old were divided into three groups. The 1st reversion of virulence trials used 105. 0TCID50/mL neck intramuscular injection of IBRV LNM attenuated vaccine strain. Then,the nose swab samples were collected for continuous 14 days. After passed through 0. 45 μm filter membrane,nasal swabs mixture was prepared as the virulence test inoculum for next generation. The body temperature was detected and clinical observation was carried out for continuous 14 days after inoculation. The inoculation dose was 1ml / cattle. Blood was collected on the 0 and 14 thdays of animal vaccination. After serum isolation,it was used for the antibody detection of serum. Research results showed that no virus was isolated from the nasal swabs from the F2 generation; vaccinated animals did not show any clinical signs of IBR; serological testing of IBRV antibody was negative,which indicated that the strain-inoculated animals did had reversion of virulence in all three generations.

A dog carrying mutations in AVP-NPII exhibits key features of central diabetes insipidus

Central diabetes insipidus(CDI)is a rare disease characterized by the excretion of copious amounts of diluted urine(polyuria),excess water intake(polydipsia),and a rise in serum sodium concentration(hypernatremia)(Christ-Crain et al.,2019).The neuropeptide arginine-vasopressin(AVP)is synthesized as a preprohormone along with its carrier protein neurophysin II(NPII)in hypothalamic supraoptic(SON)and paraventricular(PVN)magnocellular neurons,stored in the posterior pituitary,and secreted into the circulation.

Optimal Timing of a Commonly-Used Rabies Virus for Neural Recording and Manipulation

Dear Editor,A fundamental goal of modern neuroscience is to dissect the neural circuits in the brain and understand their functions.As a powerful tool for anatomical studies,the genetically modified rabies virus(RV)SADAG(EnvA)has achieved great success in mapping presynaptic inputs to genetically marked neurons[1-3].

Gα_s Relays Sphingosine-1-Phosphate Receptor 1 Signaling to Stabilize Vascular Endothelial-Cadherin at Endothelial Junctions to Control Mouse Embryonic Vascular Integrity

Sphingosine-1-phosphate receptor 1 （S1PR1）, a G protein-coupled recep （GPCR）. controls vasct stability by stabilizing vascular endothelial （VE）-cadherin junctional localization and inhibiting vascular endothelial growth factor receptor 2（VEGFR2） signaling. However, the molecular mechanisms that link S1PR1 signaling to intracellular effectors remain unknown.In this study,we demonstrate that the heterotrimeric G protein subfamily member Gαs, encoded by GNAS,acts as a relay mediator of S1PR1 signaling to control vascular integrity by stabilizing VE-cadherin at endothelial junctions. The endothelial cell -spectific deletion of Gαs in mice causes early embryonic lethality with massive hemorrhage and a disorganized Vaseuiature.The immunostaining results revealed that Gαs deletion remarkably reduces the junctional localization of VE-cadherin, whereas the mull cell coverage of the vessels is not impaired.In addition, we found-that Gαs depletion blocks the S1PR1-activation induced VE-cadherin stabilization at junctons,supporting that Gαs acts downstream of S1PR1 signaling ThuS, our results demonstrate that Gαs is an essential mediator to relay S1PR1 signaling and maintain vascular integrity.

Polymorphisms of chemokine receptors and its ligand alleles influencing genetic suscepti-bity to HIV-1 infection in eight ethnic groups in Chinese mainland

Limited genetic information is available concerning the polymorphisms of HIV-1 resistant genes in indigenous Chinese populations. The aim of this study is to identify the allelic frequencies of the chemokine and chemokine receptor genes in the Chinese mainland. Genomic DNA samples extracted from whole blood of 2318 subjects were analyzed by using PCR or PCR/restriction fragment length polymorphism (RFLP) assays, and further confirmed by direct DNA sequencing. Higher frequencies of mutant CCR2-64I (19.15%-28.79%) and SDF1-3’A (19.10% -29.86%) alleles were found in subjects of 8 ethnic groups in the Chinese mainland. In contrast, the △32 mutation in CCRS gene occurs at a very low frequency (0.0016, n=1287) in Han population. A relatively high frequency of CCR5-wt/△32 heterozygotes was observed in Uygurian and Mongolian populations. No A32 mutation allele was detected in Tibetan and other 4 ethnic groups in Yunnan Province. There was no CCR5-m303 mutation in subjects of any ethnic group in the Chinese

Effects of dietary copper on elemental balance,plasma minerals and serum biochemical parameters of growing-furring male mink(Mustela vison)

The objectives of this study were to study the effects of different levels of dietary copper on copper and zinc balance, plasma minerals and serum biochemical parameters of mink in the growing-furring periods. One hundred and five standard dark male mink were randomly assigned to seven groups with the following dietary treatments: basal diet with no supplemental Cu(Control); basal diet supplemented with either 6,12, 24, 48, 96, or 192 mg/kg Cu from copper sulfate,respectively. The average daily gain(ADG) linearly(P = 0.0026, P = 0.0006) responded to increasing levels of Cu; maximal growth was seen in the Cu24 group. Feed efficiency tended to improve with the increase of dietary copper level(linear P = 0.0010, quad, P = 0.0011). Fecal copper, urinary copper, retention copper responded in a linear(P < 0.05) fashion with increasing level of Cu. The effect of level of Cu was linear(P < 0.001) for plasma Cu concentration. The serum glutamic-oxalacetic transaminase(GOT) and glutamic-pyruvic transaminase(GPT) activities were increased linearly(P < 0.05) with dose of Cu, but serum total protein(TP) and albumin(ALB) concentrations decreased linearly(P < 0.05) as dietary copper levels increased. Effect of level of Cu was linear(P < 0.001) for serum ceruloplasmin(CER) concentration or Cu-Zn superoxide dismutase(Cu-Zn SOD) activity. Supplemental dose of Cu linearly decreased serum triglyceride(TG)(P = 0.011) and total cholesterol(TC)(P = 0.007). Our results indicated that the activity of Cu-dependent enzymes was enhanced by increasing dietary Cu concentration and that supplementation of Cu in the diet of mink could alter the plasma lipid profile and copper concentration.

USP11 regulates p53 stability by deubiquitinating p53

The p53 tumor suppressor protein coordinates the cellular responses to a broad range of cellular stresses, leading to DNA repair, cell cycle arrest or apoptosis. The stability of p53 is essential for its tumor suppressor function, which is tightly controlled by ubiquitin-dependent degradation primarily through its negative regulator mudne double minute 2 （Mdm2）. To better understand the regulation of p53, we tested the interaction between p53 and USP11 using co-immunoprecipitation. The results show that USP11, an ubiquitin-specific protease, forms specific complexes with p53 and stabilizes p53 by deubiquitinating it. Moreover, down-regulation of USP11 dramatically attenuated p53 in- duction in response to DNA damage stress. These findings reveal that USP11 is a novel regulator of p53, which is required for p53 activation in response to DNA damage.

Mink Circovirus Can Infect Minks, Foxes and Raccoon Dogs

Dear Editor,Mink circovirus (MiCV), which is clustered in the genus Circovirus of the family Circoviridae, was first described in minks from farms in Dalian, China in 2013 (Lian et al.2014). The complete single-stranded circular genome of the virus is 1,753 nucleotides long and contains two major open reading frames (ORFs), designated ORF1 (Rep gene)and ORF2 (Cap gene)(Lian et al. 2014;Ge et al. 2018).

Predator-induced changes of male and female mating preferences: innate and learned components

While many mati ng pref ere nces have a genetic basis, the question remai ns as to whether and how learning/experience can modify individual mate choice decisions. We used wild-caught (predator-experienced) and Fi laboratory-reared (predator-naive) invasive Western mosquitofish Gambusia affinis from China to test whether mating preferences (assessed in a first mate choice test) would change under immediate predation threat. The same individuals were tested in a second mate choice test during which 1 of 3 types of animated predators was presented: 1) a co-occurring predator, 2) a co-evolved but not currently co-occurring predator, and 3) a non-piscivorous species as control. We compared preference scores derived from both mate choice tests to separate innate from experiential effects of predation. We also asked whether predator-induced changes in mating preferences would differ betwee n sexes or depend on the choosing individual's personality type and/or body size. Wild-caught fish altered their mate choice decisions most when exposed to the co-occurring predator whereas laboratory-reared individuals responded most to the co-evolved predator, suggesting that both innate mechanisms and learning effects are involved. This behavior likely reduces individuals' risk of falling victim to predation by temporarily moving away from high-quality (i.e., conspicuous) mating partners. Accordingly, effects were stronger in bolder than shyer, large- compared with small-bodied, and female compared with male focal individuals, likely because those phenotypes face an increased predation risk overall. Our study adds to the growing body of literature appreciating the complexity of the mate choice process, where an array of intrinsic and extrinsic factors interacts during decision-making.

An Attenuated Highly Pathogenic Chinese PRRS Viral Vaccine Confers Cross Protection to Pigs against Challenge with the Emerging PRRSV NADC30-Like Strain

A novel PRRSV strain was isolated in China that was genetically similar to the NADC30 strain which is reported to have spread throughout China. The objective of the present study was to evaluate the cross-protective efficacy of the live vaccine TJM-F92 in young pigs against challenge with a NADC30-like strain, HN201605. Twenty-five PRRSV-and antibody-free pigs were randomly divided into the following five groups: Vac/ChA, Unvac/ChA, Vac/ChB, Unvac/ChB and the mock.The pigs in groups Vac/ChA and Vac/ChB were inoculated intramuscularly with 1 mL TJM-F92(10^(5.0) TCID_(50)/mL). At28 days post vaccination(0 days post challenge), groups Vac/ChA and Unvac/ChA were inoculated intranasally with 10^(4.5) TCID_(50)/mL PRRSV strain TJ F3(2 mL/pig), while groups Vac/ChB and Unvac/Ch B were inoculated, using the same route, with the same dose of the NADC30-like strain HN201605 F3. Protective effects of the PRRSV strain were observed in all pigs in the Vac/ChA and Vac/ChB groups. Neither high fever nor signs of clinical disease were observed through the experiment in these groups, whereas pigs in Unvac/ChA group exhibited serious clinical symptoms, pathological lesions,and weight loss. In Unvac/ChB group, pigs developed milder clinical symptoms, which demonstrated that the NADC30-like strain HN201605 had moderate pathogenicity. The results suggest that the MLV vaccine strain TJM-F92 is an effective and safe vaccine candidate for use in China.

Comparing the biological coherence of network clusters identified by different detection algorithms

Protein-protein interaction networks serve to carry out basic molecular activity in the cell. Detecting the modular structures from the protein-protein interaction network is important for understanding the organization, function and dynamics of a biological system. In order to identify functional neighbor- hoods based on network topology, many network cluster identification algorithms have been devel- oped. However, each algorithm might dissect a network from a different aspect and may provide dif- ferent insight on the network partition. In order to objectively evaluate the performance of four com- monly used cluster detection algorithms: molecular complex detection (MCODE), NetworkBlast, shortest-distance clustering (SDC) and Girvan-Newman (G-N) algorithm, we compared the biological coherence of the network clusters found by these algorithms through a uniform evaluation framework. Each algorithm was utilized to find network clusters in two different protein-protein interaction net- works with various parameters. Comparison of the resulting network clusters indicates that clusters found by MCODE and SDC are of higher biological coherence than those by NetworkBlast and G-N algorithm.