Antheraea pernyi nucleopolyhedrovirus (ApNPV) PstI-B and C fragments were cloned and sequenced. ApNPV PstI-B was 7406 bp long, contained seven open reading frames (orfs)/genes, including p87, he65, pnk/pnl, odv-ec43 and Orgyia pseudotsugata multicapsid nucleopolyhedrovirus (OpMNPV) orf107,orf108 homologue on either strands of genomic DNA. ApNPV PstI-C was 6663 bp long, contained eleven orfs/genes, including pk-1, orf1629, polh, lef-2, ptp-2, ctl-1, ptp-1 and OpMNPV orf5, orf7, orf8, orf1 1 homologue on either strands of genomic DNA. Among the eighteen baculovirus genes identification, he65 and orf1629 were two diverse genes, while polh and lef-2 were two conserved genes. ApNPV was the third baculovirus found to contain pnk/pnl gene, the fourth baculovirus found to contain both ptp-1 and ptp-2 gene.

Objective To evaluate the application of time to positivity (TTP)in differential diagnosis of intracranial infection caused by coagulase-negative Staphylococcus (CNS).Methods One hundred and twenty-four adult patients with positive CNS isolated from cerebrospinal fluid (CSF)including 70 cases with intracranial infection and 54 cases of CSF contamination,who were admitted in Xiangya Hospital of Central South University during January to December 201 5,were retrospectively analyzed.The difference of TTP between two groups was compared,receiver operating characteristic (ROC)curve was analyzed and the area under the ROC curve (AUC)was calculated.The application of TTP in differential diagnosis of CNS infection was evaluated.SPSS 1 8.0 software was used to analyze the data.Results TTP in intracranial infection group was shorter than that in CSF contamination group [(23.5 ±7.5 )h vs. (37.6 ±1 0.5)h,t =-8.71 7,P =0.000].The AUC of TTP was 0.854.Taking the cut-off value of 27.94 h,the sensitivity,specificity,positive and negative predictive values in differentiation of two groups were 72.7%,91 .4%,90.0% and 72.2%,respectively.There were statistically differences in TTP of Staphylococcus epidermidis,Staphylococcus haemolyticus and Staphylococcus capitis between two groups (Z =-4.496,-2.322 and -2.399,respectively,P <0.05 or <0.01 ).Conclusion TTP can be used to discriminate early intracranial infection and CSF contamination caused by CNS,and also can identify intracranial infection caused by different categories of CNS.

Innovative and practical platform safety management is the foundation of the innovation and entrepreneurship for undergraduate students in universities and colleges, which plays a critical role in maintaining the stability of the university and society, and ensuring the safety of teachers and students. In this present paper, the complex situation and problems of innovative and practical platform safety management, such as the imperfect regulations, weak atmosphere, incomplete facilities, unprofessional management, etc., are discussed under the background of "innovation and entrepreneurship" in China. With the combination of the above problems and the actual situation of Chengdu Medical College, the constructions of the regulations, such as access regulation, certification regulation, check regulation, rewards and punishment regulation, responsibility regulation, and a series of standard operating procedures (SOP) have also been carried out. Furthermore, adequate safety education and proper safety drills have been performed during the whole process of safety management. In addition, a diverse and professional management team with putting more emphasis on the development of safety facilities has been formed. In conclusions, the practice and exploration on the safety management of undergraduate students' innovative and practical platform in this paper may provide a theoretical basis and practical value to optimize the safety management in local universities and colleges under the background of "innovation and entrepreneurship" in China.

Objective To investigate and analyze the epidemiological and pathogenic characteristics on a case of human Streptococcus suis type 2 infection in Minhang District, Shanghai, and to provide evidence for early warning and prevention and control measures of rare and imported zoonotic acute infectious diseases in Shanghai. Methods By inquiring the patient medical history and epidemiological history and on-site environmental investigation, the infection route and source of the case were examined. The pathogenic culture of cerebrospinal fluid (CSF) was used to isolate Streptococcus suis, and Vitek2GP was used to identify the isolated strains. The PCR technique was used to detect species specific genes and virulence genes. Results The clinical manifestations of the patient were high fever with headache, nausea, vomiting and stiff neck. Blood tests showed a significant increase in c-reactive protein, an increase in lymphocyte percentage, and a decrease in platelet count. Head CT examination showed bilateral ethmoidal sinus and bilateral maxillary sinus inflammation, and significantly increased CSF white blood cell count and immunoglobulin. The case's CSF sample was positive for species specific genes (16SrRNA) and 2 virulence genes (cps-2j and ef). Conclusion This case was human Streptococcus suis type 2 with meningitis symptoms. Good prognosis was associated with timely diagnosis and treatment as well as the types of virulence factors. Medical institutions should identify early infection and take timely treatment as soon as possible to avoid severe illness and death cases. Departments of agriculture, health, market management, and others should consummate the reporting mechanism of animal epidemic situation, and establish necessary active sentinel monitoring.

OBJECTIVE: To investigate the interventional effect of the Chinese herbal preparation Xi Fu Pai Chen(XFPC) on pulmonary inflammation and fibrosis in rats with silicosis. METHODS: A total of 144 adult specific pathogen free male SD rats were randomly divided into 6 groups: blank control group, silicosis model group, drug administration control group and groups of low-dose,medium-dose and high-dose XFPC, with 24 rats in each group. Lung silicosis model was established by single inhalation tracheal instillation method, which was treated with 50.0 g/L silica suspension, in groups except in the blank control group. On the 7 th day of modeling, the rats in the drug administration control group were orally given tetrandrine(5 mg/kg body weight), while those in the low-, medium-and high-dose groups were given 43, 86 and 192 g/L of XFPC by atomization inhalation once a day for 20 minutes, 5 days a week for 4 weeks. At the end of drug administration, the histopathological changes of the lung were observed. The number and classification of cells in bronchoalveolar lavage fluid(BALF)were examined, and the levels of malondialdehyde(MDA) and interferon-gamma(IFN-γ) in BALF were measured by enzyme-linked immunosorbent assay. RESULTS: On the 7 th day after modeling, the body weight in the drug administration control group and XFPC high-dose group decreased compared with the blank control group(P<0.05). On the 35 th day after modeling, the body weights of rats in the other 5 groups were lower than that in the blank control group(P<0.05). The pathological changes of lung tissue(infiltration of inflammatory cells, fibrosis and size of silicon nodule) in drug administration control group and XFPC low-dose group were better than those in silicosis model group by naked eyes and under light microscope. The lung coefficient, the proportion of neutrophils and the level of MDA and IFN-γ in BALF of the drug administration control group and XFPC low-dose group decreased(P<0.05), and the proportion of macrophages in BALF increased(P<0.05) compared with the silicosis model group. There was no significant difference in lung coefficients and the relevant indices of BALF between XFPC medium-, high-dose groups and silicosis model group(P>0.05). CONCLUSION: Low dosage XFPC can improve pulmonary fibrosis and inflammation in rats with silicosis, and its mechanism of action may be related to reducing the levels of IFN-γ and MDA in BALF.

The CRISPR/Cas9 gene editing system has been widely used in basic research, gene therapy and genetic engineering due to its high efficiency, fast speed and convenience. Meanwhile, the discovery of novel CRISPR/Cas systems in the microbial community also accelerated the emergence of novel gene editing tools. CRISPR/Cpf1 is the second type (V type) CRISPR system that can edit mammalian genome. Compared with the CRISPR/Cas9, CRISPR/Cpf1 can use 5'T-PAM rich region to increase the genome coverage, and has many advantages, such as sticky end of cleavage site and less homologous recombination repair. Here we constructed three CRISPR/Cpf1 (AsCpf1, FnCpf1 and LbCpf1) expression vectors in silkworm cells. We selected a highly conserved BmHSP60 gene and an ATPase family BmATAD3A gene to design the target gRNA, and constructed gHSP60-266 and gATAD3A-346 knockout vectors. The efficiency for editing the target genes BmATAD3A and BmHSP60 by AsCpf1, FnCpf1 and LbCpf1 were analyzed by T7E1 analysis and T-clone sequencing. Moreover, the effects of target gene knockout by different gene editing systems on the protein translation of BmHSP60 and BmATAD3A were analyzed by Western blotting. We demonstrate the CRISPR/Cpf1 gene editing system developed in this study could effectively edit the silkworm genome, thus providing a novel method for silkworm gene function research, genetic engineering and genetic breeding.
Animals ; Bombyx/metabolism* ; CRISPR-Cas Systems/genetics* ; Endonucleases/genetics* ; Gene Editing ; RNA, Guide/genetics*

Bombyx mori is a lepidopteran insect with important economic value. Bombyx mori nucleopolyhedrovirus (BmNPV) causes huge economic loss to silkworm industry in China every year. The objective of this study is to determine the anti-BmNPV mechanism of Bombyx mori strain NC99R, and to provide a basis for understanding the molecular mechanism of the silkworm resistance strain. The normal control Dazao (DZ) strain and the NC99R resistant strain were fed with occlusion bodies (OB). The median lethal dose (LD50) analysis of the DZ and NC99R showed that the LD50 of DZ was 1.2×10⁵ OBs/larva, while NC99R was 1.8×10⁶ OBs/larva. The LD50 of the NC99R was about 15 times higher than the DZ. The mortality of DZ and NC99R were analyzed, which were fed with 1×10⁶ OBs/larva and injection with 1×10⁶ BVs/larva. The results showed that the death peak of DZ was concentrated in the 4th to 6th day. And the death peak of NC99R was concentrated in the 6th to 8th day, with a delay of 1-2 days compared with the control. The BmNPV DNA copy number showed that the BmNPV genome in DZ proliferated rapidly. The copy number of BmNPV DNA in NC99R were increased slowly after oral infection and body injection. HE staining showed that midgut tissue has no significant difference between DZ and NC99R in the early stage of oral infection. At 96 h p.i., the nucleus of DZ midgut became larger and shedding. The NC99R had enlarged nuclei, but the cells were still arranged neatly. Finally, the expression of virus genes in different periods were analyzed by RT-PCR. The results indicated that the immediate early gene ie-1 expression levels began to down-regulate after 24 h p.i.. The early, late, and extremely late genes were also down-regulated, and finally maintained at a lower expression level.

ObjectiveThis study aimed to understand the epidemiological characteristics of hand-foot-mouth disease （HFMD） in Minhang District， Shanghai from 2009 to 2020， and provide a scientific basis for the prevention and control of HFMD. MethodsThe case information of HFMD was collected from the National Notifiable Infectious Diseases Reporting System of Chinese Center for Disease Control and Prevention. We used descriptive epidemiological methods to analyze the population characteristics， temporal and spatial distribution of HFMD， the pathogen composition of the case and its changing trend. ResultsFrom 2009 to 2020， a total of 66，198 cases of HFMD were reported in Minhang District， Shanghai， including 377 severe cases （severe case rate 0.57%） and 3 deaths （severs case fatality rate 0.80%）. There were more cases of HFMD in boys than in girls （1.5∶1）. HFMD patients aged under 5 years predominated， accounting for 88.91% of all cases. Majority of the cases （91.42%） were in scattered children （55.80%） and children in kindergartens （35.62%）. The incidence showed a cyclical trend， with low incidence years and high incidence years appearing alternately. The peak period was from April to July， and sometimes there were small peaks during October to December. A total of 12 years time-space scanning analysis revealed 3 clusters. The cluster centers were located in Wujing Town， Huacao Town and Xinzhuang Town， respectively. The proportion of EV71 in common cases was generally decreasing， and reduced to zero in 2019. The proportion of CoxA6 had increased year by year， and reached 75.00% in 2020. CoxA6 became the dominant pathogen in recent years. The number of severe cases had decreased year by year since 2010， and the dominant pathogen was EV71 （90.03% on average） in severe cases. ConclusionThe incidence of HFMD in Minhang District of Shanghai has a downward trend from 2014. The dominant pathogen changes from EV71 to CoxA6， and the dominant pathogen in severe cases is EV71. The discovered temporal and spatial clustering pattern is helpful for in-depth understanding of the distribution and epidemic trend of HFMD in Minhang District， and provides a scientific basis for epidemic prevention and control.

Here, we report a previously unrecognized syndromic neurodevelopmental disorder associated with biallelic loss-of-function variants in the RBM42 gene. The patient is a 2-year-old female with severe central nervous system (CNS) abnormalities, hypotonia, hearing loss, congenital heart defects, and dysmorphic facial features. Familial whole-exome sequencing (WES) reveals that the patient has two compound heterozygous variants, c.304C>T (p.R102*) and c.1312G>A (p.A438T), in the RBM42 gene which encodes an integral component of splicing complex in the RNA-binding motif protein family. The p.A438T variant is in the RRM domain which impairs RBM42 protein stability in vivo. Additionally, p.A438T disrupts the interaction of RBM42 with hnRNP K, which is the causative gene for Au-Kline syndrome with overlapping disease characteristics seen in the index patient. The human R102* or A438T mutant protein failed to fully rescue the growth defects of RBM42 ortholog knockout ΔFgRbp1 in Fusarium while it was rescued by the wild-type (WT) human RBM42. A mouse model carrying Rbm42 compound heterozygous variants, c.280C>T (p.Q94*) and c.1306_1308delinsACA (p.A436T), demonstrated gross fetal developmental defects and most of the double mutant animals died by E13.5. RNA-seq data confirmed that Rbm42 was involved in neurological and myocardial functions with an essential role in alternative splicing (AS). Overall, we present clinical, genetic, and functional data to demonstrate that defects in RBM42 constitute the underlying etiology of a new neurodevelopmental disease which links the dysregulation of global AS to abnormal embryonic development.
Female ; Animals ; Mice ; Humans ; Child, Preschool ; Intellectual Disability/genetics* ; Heart Defects, Congenital/genetics* ; Facies ; Cleft Palate ; Muscle Hypotonia