Deep Vein Thrombosis in Hospitalized Patients With COVID-19 in Wuhan, China: Prevalence, Risk Factors, and Outcome.

BACKGROUND: To investigate deep vein thrombosis (DVT) in hospitalized patients with coronavirus disease 2019 (COVID-19), we performed a single institutional study to evaluate its prevalence, risk factors, prognosis, and potential thromboprophylaxis strategies in a large referral and treatment center. METHODS: We studied a total of 143 patients with COVID-19 from January 29, 2020 to February 29, 2020. Demographic and clinical data, laboratory data, including ultrasound scans of the lower extremities, and outcome variables were obtained, and comparisons were made between groups with and without DVT. RESULTS: Of the 143 patients hospitalized with COVID-19 (age 63±14 years, 74 [51.7%] men), 66 patients developed lower extremity DVT (46.1%: 23 [34.8%] with proximal DVT and 43 [65.2%] with distal DVT). Compared with patients who did not have DVT, patients with DVT were older and had a lower oxygenation index, a higher rate of cardiac injury, and worse prognosis, including an increased proportion of deaths (23 [34.8%] versus 9 [11.7%]; P=0.001) and a decreased proportion of patients discharged (32 [48.5%] versus 60 [77.9%]; P<0.001). Multivariant analysis showed an association only between CURB-65 (confusion status, urea, respiratory rate, and blood pressure) score 3 to 5 (odds ratio, 6.122; P=0.031), Padua prediction score ≥4 (odds ratio, 4.016; P=0.04), D-dimer >1.0 µg/mL (odds ratio, 5.818; P<0.014), and DVT in this cohort, respectively. The combination of a CURB-65 score 3 to 5, a Padua prediction score ≥4, and D-dimer >1.0 µg/mL has a sensitivity of 88.52% and a specificity of 61.43% for screening for DVT. In the subgroup of patients with a Padua prediction score ≥4 and whose ultrasound scans were performed >72 hours after admission, DVT was present in 18 (34.0%) patients in the subgroup receiving venous thromboembolism prophylaxis versus 35 (66.0%) patients in the nonprophylaxis group (P=0.010). CONCLUSIONS: The prevalence of DVT is high and is associated with adverse outcomes in hospitalized patients with COVID-19. Prophylaxis for venous thromboembolism may be protective in patients with a Padua protection score ≥4 after admission. Our data seem to suggest that COVID-19 is probably an additional risk factor for DVT in hospitalized patients.

[Effect of Yunnan herb Laggera pterodonta against influenza A (H1N1) virus in vitro].

Laggera pterodonta is commonly used for treating influenza in Southwest China, especially in Yunnnan province. The main clinical effects of L. pterodonta include anti-influenza, anti-microbial, anti-inflammatory. To investigate the anti-influenza A (H1N1) virus effect of L. pterodonta, neutralization inhibition and proliferation inhibition tests were performed. MDCK culture method was used to observe the cytopathic effect (CPE) of extracts from L. pterodonta in inhibiting influenza A (H1N1) virus and haemagglutination titre of H1N1 virus in vitro. The culture medium were collected at 24 h, 48 h, 72 h, 96 h, and detected by Real time RT-PCR, in order to compare the effect of different extracts from L. pterodonta on in vitro proliferation of H1N1, virus. The result of neutralization inhibition test showed that hemagglutination titer of ethyl acetate extract were 8 times lower at 72 h; in proliferation inhibition test, hemagglutination titer of ethyl acetate extracts reduced by 2 and 4 times. According to the results of Real time RT-PCR test, the H1N1 inhibition ratio of ethyl acetate extract was 72.5%, while the proliferation inhibition ratio of ethyl acetate extract was 25.3%; as for petroleum ether extracts, the H1N1 inhibition ratio was 60.2%, while the proliferation inhibition ratio was 81.4%. In conclusion, both ethyl acetate extract and petroleum ether extract of L. pterodonta have significant neutralization and direct proliferation inhibition effects on influenza A virus.

Characterization of a nodavirus replicase revealed a de novo initiation mechanism of RNA synthesis and terminal nucleotidyltransferase activity.

Nodaviruses are a family of positive-stranded RNA viruses with a bipartite genome of RNAs. In nodaviruses, genomic RNA1 encodes protein A, which is recognized as an RNA-dependent RNA polymerase (RdRP) and functions as the sole viral replicase protein responsible for its RNA replication. Although nodaviral RNA replication has been studied in considerable detail, and nodaviruses are well recognized models for investigating viral RNA replication, the mechanism(s) governing the initiation of nodaviral RNA synthesis have not been determined. In this study, we characterized the RdRP activity of Wuhan nodavirus (WhNV) protein A in detail and determined that this nodaviral protein A initiates RNA synthesis via a de novo mechanism, and this RNA synthesis initiation could be independent of other viral or cellular factors. Moreover, we uncovered that WhNV protein A contains a terminal nucleotidyltransferase (TNTase) activity, which is the first time such an activity has been identified in nodaviruses. We subsequently found that the TNTase activity could function in vitro to repair the 3' initiation site, which may be digested by cellular exonucleases, to ensure the efficiency and accuracy of viral RNA synthesis initiation. Furthermore, we determined the cis-acting elements for RdRP or TNTase activity at the 3'-end of positive or negative strand RNA1. Taken together, our data establish the de novo synthesis initiation mechanism and the TNTase activity of WhNV protein A, and this work represents an important advance toward understanding the mechanism(s) of nodaviral RNA replication.

The nonstructural protein 2C of a Picorna-like virus displays nucleic acid helix destabilizing activity that can be functionally separated from its ATPase activity.

Picorna-like viruses in the Picornavirales order are a large group of positive-strand RNA viruses that include numerous important pathogens for plants, insects, and humans. In these viruses, nonstructural protein 2C is one of the most conserved proteins and contains ATPase activity and putative RNA helicase activity. Here we expressed 2C protein of Ectropis obliqua picorna-like virus (EoV; genus Iflavirus, family Iflaviridae, order Picornavirales) in a eukaryotic expression system and determined that EoV 2C displays ATP-independent nucleic acid helix destabilizing and strand annealing acceleration activity in a concentration-dependent manner, indicating that this picornaviral 2C is more like an RNA chaperone than like the previously predicted RNA helicase. Our further characterization of EoV 2C revealed that divalent metal ions, such as Mg(2+) and Zn(2+), inhibit 2C-mediated helix destabilization to different extents. Moreover, we determined that EoV 2C also contains ATPase activity like that of other picornaviral 2C proteins and further assessed the functional relevance between its RNA chaperone-like and ATPase activities using mutational analysis as well as their responses to Mg(2+). Our data show that, when one of the two 2C activities was dramatically inhibited or almost abolished, the other activity could remain intact, showing that the RNA chaperone-like and ATPase activities of EoV 2C can be functionally separated. This report reveals that a picorna-like virus 2C protein displays RNA helix destabilizing and strand annealing acceleration activity, which may be critical for picornaviral replication and pathogenesis, and should foster our understanding of picorna-like viruses and viral RNA chaperones.

Elective cesarean delivery as a predisposing factor of respiratory syncytial virus bronchiolitis in children.

OBJECTIVE: To investigate the effect of cesarean delivery and other predisposing factors of respiratory syncytial virus (RSV)-positive acute bronchiolitis in children. MATERIAL AND METHOD: The case-control study was conducted in three main tertiary hospitals in Kunming, China between September 2012 and July 2013. Children with first episode of wheezing diagnosed as bronchiolitis and testedfor RSV were included RSV was detected by real-time reverse transcription polymerase chain reaction. Mode ofdelivery and characteristics of children, parents, and household were interviewed and analyzed with RSV-positive status by multiple logistic regression. RESULTS: Of 265 children, RSV-positive was found in 75.5%, and the majority of children (83.3%) were younger than 12 months. Compared to vaginal delivery, the odds of RSV-positive detection were double in children born by elective cesarean delivery (adjusted odds ratio 2.32; 95% confidence interval 1.19-4.52). Children aged less than 6 months, born in the rainy season, having maternal history of asthma and living in family that smoked more than 20 cigarettes per day were more likely to be RSV-positive. CONCLUSION: Children born by elective cesarean delivery increased the risk of RSV-positive acute bronchiolitis after adjusting for age, birth season, maternal asthma, and family smoking status.

Ben-JNK signaling is required for host mortality during Periplaneta fuliginosa densovirus infection.

BACKGROUND: Cockroaches are widely acknowledged as significant vectors of pathogenic microorganisms. The Periplaneta fuliginosa densovirus (PfDNV) infects the smoky-brown cockroach P. fuliginosa and causes host mortality, which identifies the PfDNV as a species-specific and environmentally friendly biopesticide. However, although the biochemical characterization of PfDNV has been extensively studied, the immune response against PfDNV remains largely unclear. RESULTS: Here, we investigated the replication of PfDNV and its associated pathological phenotype in the foregut and hindgut. Consequently, we dissected and performed transcriptome sequencing on the foregut, midgut, and hindgut separately. We revealed the up-regulation of immune response signaling pathway c-Jun N-terminal kinase (JNK) and apoptosis in response to viral infection. Furthermore, knockdown of the JNK upstream gene Ben resulted in a decrease in virus titer and delayed host mortality. CONCLUSION: Taken together, our findings provide evidence that the Ben-JNK signaling plays a crucial role in PfDNV infection, leading to excessive apoptosis in intestinal tissues and ultimately resulting in the death of the host. Our results indicated that the host response to PfDNV fosters viral infection, thereby increasing host lethality. This underscores the potential of PfDNV as a viable, environmentally friendly biopesticide. © 2024 Society of Chemical Industry.

Revisiting current "barefoot doctors" in border areas of China: system of services, financial issue and clinical practice prior to introducing integrated management of childhood illness (IMCI).

BACKGROUND: Under-5-years child mortality remains high in rural China. Integrated management of childhood illness (IMCI) was introduced to China in 1998, but only a few rural areas have been included. This study aimed at assessing the current situation of the health system of rural health care and evaluating the clinical competency of village doctors in management of childhood illnesses prior to implementing IMCI programme in remote border rural areas. METHODS: The study was carried out in the border areas of Puer prefecture of Yunnan province. There were 182 village doctors in the list of the health bureau in these border areas. Of these, 154 (84.6%) were recruited into the study. The local health system components were investigated using a qualitative approach and analyzed with triangulation of information from different sources. The clinical component was assessed objectively and quantitatively presented using descriptive statistics. RESULTS: The study found that the New Rural Cooperative Medical Scheme (NRCMS) coordinated the health insurance system and the provider service through 3 tiers: village doctor, township and county hospitals. The 30 RMB per person per year premium did not cover the referral cost, and thereby decreased the number of referrals. In contrast to available treatment facilities and drug supply, the level of basic medical education of village doctors and township doctors was low. Discontent among village doctors was common, especially concerning low rates of return from the service, exceptions being procedures such as injections, which in fact may create moral hazards to the patients. Direct observation on the assessment and management of paediatric patients by village doctors revealed inadequate history taking and physical examination, inability to detect potentially serious complications, overprescription of injection and antibiotics, and underprescription of oral rehydration salts and poor quality of counseling. CONCLUSION: There is a need to improve health finance and clinical competency of the village doctors in the study area.

Evaluation of short term integrated management of childhood illness training on the clinical competency of village doctors in Yunnan, China.

OBJECTIVE: Evaluate the clinical competency in detection and management complicated pneumonia, diarrhea, and measles cases of trainees who were village doctors in Puer city of Yunnan province, China. MATERIAL AND METHOD: One hundred fifty four village doctors working in the border areas were trainedfor 5-day Integrated Management of Childhood Illness (IMCI) training, which was adapted to World Health Organization (WHO) guidelines. Pre- and post-training assessment based on a modified WHO package was carried out. RESULTS: Statistically significant improvements in almost allperformances were observed. However, the level of post-training achievement was still low. Correct classifications of severe pneumonia, measles, and diarrhea were found in 63.0% 54.3% and 83.5%, respectively in the post-training period CONCLUSION: The training package needs to be further strengthened.

Tumor elimination by clustered microRNAs miR-306 and miR-79 via noncanonical activation of JNK signaling.

JNK signaling plays a critical role in both tumor promotion and tumor suppression. Here, we identified clustered microRNAs (miRNAs) miR-306 and miR-79 as novel tumor-suppressor miRNAs that specifically eliminate JNK-activated tumors in Drosophila. While showing only a slight effect on normal tissue growth, miR-306 and miR-79 strongly suppressed growth of multiple tumor models, including malignant tumors caused by Ras activation and cell polarity defects. Mechanistically, these miRNAs commonly target the mRNA of an E3 ubiquitin ligase ring finger protein 146 (RNF146). We found that RNF146 promotes degradation of tankyrase (Tnks), an ADP-ribose polymerase that promotes JNK activation in a noncanonical manner. Thus, downregulation of RNF146 by miR-306 and miR-79 leads to hyper-enhancement of JNK activation. Our data show that, while JNK activity is essential for tumor growth, elevation of miR-306 or miR-79 overactivate JNK signaling to the lethal level via noncanonical JNK pathway and thus eliminate tumors, providing a new miRNA-based strategy against cancer.

Emotional Dysregulation and Time Structure Mediate the Link between Perceived Stress and Insomnia among Unemployed Young People in China: A Cross-Sectional Study.

In recent years, the problem of youth unemployment in China has become a great concern. Unemployed young people often find themselves under tremendous stress and vulnerable to sleep problems. The link between perceived stress and insomnia has been widely documented, but the potential mechanisms underlying this relationship have not been thoroughly revealed. The objective of this study was to examine the underlying mechanisms linking perceived stress to insomnia through testing the mediating effects of emotional dysregulation and time structure. Through a multiple-stage convenience sampling, 511 unemployed young people (38.6% women; mean age = 21.51; SD = 2.22) were measured using the 10-item Perceived Stress Scale (PSS-10), Insomnia Severity Index (ISI), 16-item Difficulties in Emotion Regulation Scale (DERS-16), and Time Structure Questionnaire (TSQ). Based on the statistical description of the sample, chi-squared tests, bivariate correlation analyses, and mediation analyses were performed. The study indicated that 53.0% (95% CI: 48.7-57.4%) of the participants reported probable insomnia, and both insomnia and perceived stress demonstrated strong associations with emotional dysregulation and time structure, which served as partial mediators between perceived stress and insomnia symptoms according to mediation analyses. Path analysis further revealed that, after controlling for age and gender, emotional dysregulation and time structure accounted for 31.8 and 17.6% of the effect of perceived stress on insomnia, respectively. This study provides empirical support for the association among perceived stress, emotional dysregulation, time structure, and insomnia symptoms. To improve the sleep quality and general wellbeing of unemployed young people, emotional dysregulation and time structure must be taken into consideration.

Sensory stimulation program improves developments of preterm infants in Southwest China: A randomized controlled trial.

Preterm infants are prone to growth and developmental delay, especially social-emotional development. Sensory stimulation may benefit developmental outcomes for these vulnerable infants. This study aims to determine whether 5-integrated sensory stimulation (5-ISS) improves preterm infant social-emotional development. A randomized, parallel trial was conducted from November 2018 to January 2020 at three tertiary hospitals in Kunming, China. Preterm infants were eligible if gestational ages were from 28 to 36 weeks based on ultrasound results when discharged from neonatal wards. Two hundred preterm infants (male n = 110, female n = 90) were randomly allocated to the 5-ISS intervention group (n = 98) and the standard care group (n = 102). Social-emotional development was assessed with the Ages and Stages Questionnaires: Social-Emotional (ASQ:SE). Temperament was assessed with the Infant Behavior Questionnaire-Revised. Anthropometry, which included weight, length, and head circumference, was measured at corrected ages of 1, 3, and 6 months. Demographic and clinical characteristics were similar between the intervention and the standard care groups. At 1- and 3-month corrected age, no significant differences between the two groups were observed in terms of infant development and temperament. At 6 months, significant disparities were found in the social-emotional development scale (mean difference -0.29, 95% CI: -0.58, < -0.001, p = 0.01), infant length (mean difference 0.70, 95% CI: < 0.001, 1.4, p = 0.03), distress to limitation (p = 0.04), and sadness (p = 0.03). A mixed model revealed that the 5-ISS intervention positively affected social-emotional development, length, distress to limitation, and sadness for preterm infants. Integrated sensory stimulation has benefits on social-emotional development, temperament, and length for preterm infants. This program provides a feasible method to promote social-emotional development for preterm infants.

Identification of a novel collagen-like peptide by high-throughput screening for effective wound-healing therapy.

Tissue regeneration and wound healing are still serious clinical complications globally and lack satisfactory cures. Inspired by the impressive regeneration ability of the post-injury earthworms and their widely accepted medicinal properties, we screened and identified a novel collagen-like peptide from the amputated earthworms using high-throughput techniques, including transcriptomics, proteomics, and mass spectrum. The identified collagen-like peptide col4a1 was cloned and expressed to comprehensively investigate the wound healing effect and underlying mechanism. It exerted significant effects on wound healing both in vitro and in vivo, including enhanced viability, proliferation, migration of fibroblasts, granulation, and collagen deposition. Moreover, the col4a1 functioned via binding with integrin α2ß1 and upregulating the RAS/MAPK signaling pathway. This work demonstrates that the novel collagen-like peptide col4a1 obtained from the amputated earthworms enables enhanced wound healing and provides new opportunities for wound care.

Pixelwise H-score: A novel digital image analysis-based metric to quantify membrane biomarker expression from immunohistochemistry images.

Immunohistochemistry (IHC) assays play a central role in evaluating biomarker expression in tissue sections for diagnostic and research applications. Manual scoring of IHC images, which is the current standard of practice, is known to have several shortcomings in terms of reproducibility and scalability to large scale studies. Here, by using a digital image analysis-based approach, we introduce a new metric called the pixelwise H-score (pix H-score) that quantifies biomarker expression from whole-slide scanned IHC images. The pix H-score is an unsupervised algorithm that only requires the specification of intensity thresholds for the biomarker and the nuclear-counterstain channels. We present the detailed implementation of the pix H-score in two different whole-slide image analysis software packages Visiopharm and HALO. We consider three biomarkers P-cadherin, PD-L1, and 5T4, and show how the pix H-score exhibits tight concordance to multiple orthogonal measurements of biomarker abundance such as the biomarker mRNA transcript and the pathologist H-score. We also compare the pix H-score to existing automated image analysis algorithms and demonstrate that the pix H-score provides either comparable or significantly better performance over these methodologies. We also present results of an empirical resampling approach to assess the performance of the pix H-score in estimating biomarker abundance from select regions within the tumor tissue relative to the whole tumor resection. We anticipate that the new metric will be broadly applicable to quantify biomarker expression from a wide variety of IHC images. Moreover, these results underscore the benefit of digital image analysis-based approaches which offer an objective, reproducible, and highly scalable strategy to quantitatively analyze IHC images.

The steroid-induced microRNA let-7 regulates developmental growth by targeting cdc7 in the Drosophila fat body.

In insects, 20-hydroxyecdysone (20E) limits systemic growth by triggering developmental transitions. Previous studies have shown that 20E-induced let-7 exhibits crosstalk with the cell cycle. Here, we examined the underlying molecular mechanisms and physiological functions of 20E-induced let-7 in the fat body, an organ for energy storage and nutrient mobilization which plays a critical role in the larval growth. First, the overexpression of let-7 decreased the body size and led to the reduction of both nucleolus and cell sizes in the larval fat body. In contrast, the overexpression of let-7-Sponge increased the nucleolus and cell sizes. Moreover, we found that cdc7, encoding a conserved protein kinase that controls the endocycle, is a target of let-7. Notably, the mutation of cdc7 in the fat body resulted in growth defects. Overall, our findings revealed a novel role of let-7 in the control of endoreduplication-related growth during larval-prepupal transition in Drosophila.

Using BI-RADS Stratifications as Auxiliary Information for Breast Masses Classification in Ultrasound Images.

Breast Ultrasound (BUS) imaging has been recognized as an essential imaging modality for breast masses classification in China. Current deep learning (DL) based solutions for BUS classification seek to feed ultrasound (US) images into deep convolutional neural networks (CNNs), to learn a hierarchical combination of features for discriminating malignant and benign masses. One existing problem in current DL-based BUS classification was the lack of spatial and channel-wise features weighting, which inevitably allow interference from redundant features and low sensitivity. In this study, we aim to incorporate the instructive information provided by breast imaging reporting and data system (BI-RADS) within DL-based classification. A novel DL-based BI-RADS Vector-Attention Network (BVA Net) that trains with both texture information and decoded information from BI-RADS stratifications was proposed for the task. Three baseline models, pre-trained DenseNet-121, ResNet-50 and Residual-Attention Network (RA Net) were included for comparison. Experiments were conducted on a large scale private main dataset and two public datasets, UDIAT and BUSI. On the main dataset, BVA Net outperformed other models, in terms of AUC (area under the receiver operating curve, 0.908), ACC (accuracy, 0.865), sensitivity (0.812) and precision (0.795). BVA Net also achieved the high AUC (0.87 and 0.882) and ACC (0.859 and 0.843), on UDIAT and BUSI. Moreover, we proposed a method that integrates both BVA Net binary classification and BI-RADS stratification estimation, called integrated classification. The introduction of integrated classification helped improving the overall sensitivity while maintaining a high specificity.

Estimation of personal ozone exposure using ambient concentrations and influencing factors.

Evidence is limited regarding whether ambient monitoring can properly represent personal ozone exposure. We conducted a longitudinal panel study to measure personal exposure to ozone using real-time personal ozone monitors. Corresponding ambient ozone concentrations and possible influencing factors (meteorological conditions and activity patterns) were also collected. We used linear mixed-effect models to analyze personal-ambient ozone concentration associations and possible influencing factors. Ambient ozone concentrations were around two to three times higher than personal ozone (43.1 µg/m3 on average) and their correlations were weak with small slopes (0.35) and marginal R square (RM2) values (0.24). Larger RM2 values were found under high temperature (>29.5 °C), low humidity (<62.1%), good ventilation conditions (>4 h) and for individuals spent longer time outdoors (>0.6 h). In final model, personal ozone exposure was positively associated with ambient concentrations and ventilation conditions, but inversely correlated with ambient temperature and humidity. The models explained >50% of personal ozone concentration variabilities. Our results highlight that ambient ozone concentration alone is not a suitable surrogate for individual exposure assessment. Meteorological conditions (temperature and humidity) and activity patterns (windows opening and outdoor activities) that affecting personal ozone exposure should be taken into account.

Molecular epidemiology study of measles viruses in Kunming area of China.

The present study assessed the variation of measles viruses (MV) and its association with clinical manifestations in patients with MV. A total of 38 pediatric patients with MV at the acute infection stage were selected and 2 ml venous blood was collected from each of them. Serum immunoglobulin M antibodies were determined by ELISA. Urine specimens were collected from 30 of the 38 patients and associated genetic structures were detected by reverse-transcription polymerase chain reaction mapping. At the same time, clinical epidemiological manifestations were collected to perform an epidemiological analysis. The MV-positive rate within the cohort determined in serum was 100%. Seven MV strains were isolated from urine specimens of 30 patients and the positive rate was 23.33%. Four MV strains were randomly selected from the 7 strains and the results revealed that they were all of the H1a genotype. In addition, there was no significant correlation between clinical manifestation of pediatric patients with measles and the genotype of the MV. In conclusion, the preponderant genotype of MV in Kunming was H1a and there was obvious nucleotide or amino acid mutation. The clinical manifestation of MV infection in pediatric patients was not associated with the MV genotype.

Efficacy and safety of Laggera pterodonta in children 3-24 months with acute bronchiolitis: a randomized controlled trial.

INTRODUCTION: Laggera pterodonta, a traditional Chinese medicine, has been commonly used in respiratory tract infections for more than hundreds of years without any randomized controlled trials to evaluate its efficacy and safety. OBJECTIVES: To evaluate the efficacy and safety of Laggera pterodonta in hospitalized children aged 3-24 months with acute bronchiolitis. METHODS: A double-blind, randomized-controlled trial was conducted in three tertiary hospitals of Kunming, China. A total of 133 acute bronchiolitis children with an initial episode of wheezing were randomly assigned to a control mixture or Laggera pterodonta mixture. All recruited patients were given three doses of the mixture every 24 h for 5 days. Clinical symptoms and responses including adverse events in both groups were assessed and laboratory tests were done at enrolment and then after 120 h. Analysis was performed based on an intention-to-treat principle. RESULTS: Significantly more hospitalized children fulfilled the discharge criteria at 96 h and 120 h in the Laggera pterodonta mixture group compared to the control group (97% vs 75.8% P < 0.001 and 98.5% vs 89.4% P = 0.03), respectively. Better responses on clinical severity score, respiratory rate, oxygen saturation, wheezing and heart rate were also detected in the Laggera pterodonta mixture group along with lower white blood cell count, platelet count and aspartate aminotransferase. Vomiting and diarrhea were more common in the control group. CONCLUSION: Laggera pterodonta mixture is effective and safe to be prescribed in hospitalized children with acute bronchiolitis.

Building a multipurpose insertional mutant library for forward and reverse genetics in Chlamydomonas.

BACKGROUND: The unicellular green alga, Chlamydomonas reinhardtii, is a classic model for studying flagella and biofuel. However, precise gene editing, such as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein (Cas9) system, is not widely used in this organism. Screening of random insertional mutant libraries by polymerase chain reaction provides an alternate strategy to obtain null mutants of individual gene. But building, screening, and maintaining such a library was time-consuming and expensive. RESULTS: By selecting a suitable parental strain, keeping individual mutants using the agar plate, and designing an insertion cassette-specific primer for library screening, we successfully generated and maintained ~150,000 insertional mutants of Chlamydomonas, which was used for both reverse and forward genetics analysis. We obtained 26 individual mutants corresponding to 20 genes and identified 967 motility-defect mutants including 10 mutants with defective accumulation of intraflagellar transport complex at the basal body. We also obtained 929 mutants defective in oil droplet assembly after nitrogen deprivation. Furthermore, a new insertion cassette with splicing donor sequences at both ends was also constructed, which increased the efficiency of gene interruption. CONCLUSION: In summary, this library provides a multifunctional platform both for obtaining mutants of interested genes and for screening of mutants with specific phenotype.

A picorna-like virus suppresses the N-end rule pathway to inhibit apoptosis.

The N-end rule pathway is an evolutionarily conserved proteolytic system that degrades proteins containing N-terminal degradation signals called N-degrons, and has emerged as a key regulator of various processes. Viruses manipulate diverse host pathways to facilitate viral replication and evade antiviral defenses. However, it remains unclear if viral infection has any impact on the N-end rule pathway. Here, using a picorna-like virus as a model, we found that viral infection promoted the accumulation of caspase-cleaved Drosophila inhibitor of apoptosis 1 (DIAP1) by inducing the degradation of N-terminal amidohydrolase 1 (NTAN1), a key N-end rule component that identifies N-degron to initiate the process. The virus-induced NTAN1 degradation is independent of polyubiquitylation but dependent on proteasome. Furthermore, the virus-induced N-end rule pathway suppression inhibits apoptosis and benefits viral replication. Thus, our findings demonstrate that a virus can suppress the N-end rule pathway, and uncover a new mechanism for virus to evade apoptosis.