Mouse models susceptible to HCoV-229E and HCoV-NL63 and cross protection from challenge with SARS-CoV-2.

Human coronavirus 229E (HCoV-229E) and NL63 (HCoV-NL63) are endemic causes of upper respiratory infections such as the "common cold" but may occasionally cause severe lower respiratory tract disease in the elderly and immunocompromised patients. There are no approved antiviral drugs or vaccines for these common cold coronaviruses (CCCoV). The recent emergence of COVID-19 and the possible cross-reactive antibody and T cell responses between these CCCoV and SARS-CoV-2 emphasize the need to develop experimental animal models for CCCoV. Mice are an ideal experimental animal model for such studies, but are resistant to HCoV-229E and HCoV-NL63 infections. Here, we generated 229E and NL63 mouse models by exogenous delivery of their receptors, human hAPN and hACE2 using replication-deficient adenoviruses (Ad5-hAPN and Ad5-hACE2), respectively. Ad5-hAPN- and Ad5-hACE2-sensitized IFNAR-/- and STAT1-/- mice developed pneumonia characterized by inflammatory cell infiltration with virus clearance occurring 7 d post infection. Ad5-hAPN- and Ad5-hACE2-sensitized mice generated virus-specific T cells and neutralizing antibodies after 229E or NL63 infection, respectively. Remdesivir and a vaccine candidate targeting spike protein of 229E and NL63 accelerated viral clearance of virus in these mice. 229E- and NL63-infected mice were partially protected from SARS-CoV-2 infection, likely mediated by cross-reactive T cell responses. Ad5-hAPN- and Ad5-hACE2-transduced mice are useful for studying pathogenesis and immune responses induced by HCoV-229E and HCoV-NL63 infections and for validation of broadly protective vaccines, antibodies, and therapeutics against human respiratory coronaviruses including SARS-CoV-2.

Efficacy and safety of EGFR-TKIs for non-small cell lung cancer: A meta-analysis of randomized controlled clinical trials.

BACKGROUND: We conducted this meta-analysis based on updated literature and research to compare the efficacy and safety of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) as treatments for patients with non-small cell lung cancer (NSCLC). METHODS: A literature search was conducted using PubMed, Embase, Medline and Web of Science databases to perform a systematic literature search based on random control trials. In these articles, EGFR-TKIs were compared with placebos, chemotherapy, or whole-brain irradiation as treatments for NSCLC. In this research, a meta-analysis of the literature was performed to produce a combined risk ratio (RR) with a 95% confidence interval (CI) for progression-free survival (PFS), overall survival (OS), and adverse events. The data were synthesized with Review Manager 5.3 software, which was used to manage the process. RESULTS: There were 15 random control trials included in the study, involving 4249 patients in total. There was evidence that EGFR-TKIs can significantly prolong OS (RR: 0.87, 95% CI: 0.75-1) and PFS (RR: 0.75, 95% CI: 0.66-0.86) in NSCLC patients. There was an increase in the incidence of adverse events after treatment with EGFR-TKI, including diarrhea (RR: 0.18, 95% CI: 0.10-0.26), infection (RR: 0.09, 95% CI: 0.02-0.16), and rash (RR: 0.37, 95% CI: 0.22-0.51). CONCLUSIONS: It has been shown that EGFR-TKIs prolong OS and PFS in patients with NSCLC. NSCLC patients may benefit from EGFR-TKIs as an important treatment option in order to prolong their survival.

[Quality evaluation of Astragali Radix through chemical pattern recognition of fingerprint by HPLC-dAD-ELSD].

OBJECTIVE: To develop an HPLC-DAD-ELSD method for detecting the fingerprint of Astragali Radix and evaluate the quality through similarity calculation and chemical pattern recognition. METHOD: Separation was performed at 25 degreeC on an Agilent Zorbax ODS C18 column(4.6 mm x250 mm,5 microm). Gradient elution was performed with the mobile phases of acetonitrile and water containing 0. 2% formic acid. The flow rate was 0. 8 mL min-1 , and sample size was 10 microL. The UV detection wavelength was set at 280 nm. The drift tube temperature for ELSD was set at 110 degreeC , and the nebulizing gas flow rate was 3.0 L min-1. The similarity calculation and chemical pattern recognition were used for fingerprint analysis. RESULT: The HPLC-DAD-ELSD method for chromatographic fingerprint of Astragali Radix showed better results of stability, precision and repeatability. The reference chromatographic fingerprint of Astragali Radix was established on the eighteen Astragali Radix samples from different sources. The results of similarity calculation were higher than 0. 83, which was in accordance with the result of chemical pattern recognition analysis. CONCLUSION: Fingerprint and chemical pattern recognition analysis could effectively distinguish Astragali Radix from different source, which could be applied to the quality control of Astragali Radix.

Storages and leaching losses of soil water dissolved CO2 and N2O on typical land use hillslopes in southeastern hilly area of China.

The dynamics of soil water dissolved CO2 and N2O are important in determining the fates of soil CO2 and N2O. However, related mechanisms and processes have been rarely revealed. In this study, storages and leaching losses of soil water dissolved CO2 and N2O were investigated on the tea garden (TG) and bamboo forest (BF) hillslopes. Soil water storage and leaching flux were simulated by the HYDRUS-3D model and the soil water dissolved CO2 and N2O concentrations were acquired by field monitoring. Results showed that the storages of soil water dissolved CO2 and N2O ranged from 1.30 to 14.86 kg C ha-1 and 0.24 to 388.99 g N ha-1 on the TG hillslope, respectively, while they ranged from 0.49 to 52.29 kg C ha-1 and 0.50 to 14.22 g N ha-1 on the BF hillslope, respectively. The annual leaching loss of soil water dissolved CO2 and N2O were 26.17 kg C ha-1 and 29.46 g N ha-1, respectively, on the TG hillslope, while they were 49.51 kg C ha-1 and 4.35 g N ha-1 on the BF hillslope, respectively. The dissolved CO2 leaching loss mainly occurred in summer, especially in July on both hillslopes. Peaks of dissolved N2O leaching loss on the TG hillslope were observed after the application of basal fertilizer, accompanying with precipitation events. Instead, peaks of dissolved N2O leaching loss on the BF hillslope were observed in summer. The main influencing factors of dissolved CO2 and N2O storages were temperature, precipitation, and fertilization, with total effects generally >0.30. However, that of the dissolved CO2 and N2O leaching losses was the precipitation, with total effects >0.57. Dissolved CO2/N2O concentration was more important than soil water storage in determining the dissolved CO2/N2O storage, while the leaching flow rate was more crucial than dissolved CO2/N2O concentration in determining the dissolved CO2/N2O leaching loss. These findings expanded our knowledge of sources and sinks of greenhouse gases on the terrestrial ecosystem.

[Clinical application of deep lamellar keratoplasty for corneal macula after viral keratitis].

OBJECTIVE: To explore the clinical safety and efficacy of deep lamellar keratoplasty for corneal macula after viral keratitis. METHODS: A total of 16 patients (16 eyes) of corneal macula after viral keratitis at our center from January 2009 to June 2011 received anti-virus eye-drops and then underwent deep lamellar keratoplasty when inflammation were dissolved or quiescent. And the corneal graft diaphaneity and corneal graft rejection reactions were observed. RESULTS: All 16 patients were recovered with a success rate of 100%. After a follow-up of 18-24 months, all corneal grafts were transparent and best-corrected visual acuity was 0.2-0.6. There was no reoccurrence during the follow-up period. After a follow-up of several months, 2 of them had rejection reaction and were controlled after treatment. No rejection reaction occurred during the long-term follow-up. CONCLUSION: As a safe and efficacious procedure, deep lamellar keratoplasty can significantly raise vision acuity with few complications.

[The visual functional therapeutic effects of deep lamellar keratoplasty and penetrating keratoplasty for fungal corneal ulcer].

OBJECTIVE: To investigate the visual functional therapeutic effects of deep lamellar keratoplasty (DLK) and penetrating keratoplasty (PK) on perforated fungal keratitis. METHODS: 64 patients (64 eyes) of fungal corneal ulcer in Ophthalmology Center of Ningbo Yinzhou People Hospital from 2004 to 2009 were retrospected, of which undergo DLK (36 patients, 36 eyes) and PK (28 patients, 28 eyes), and followed up by 12 to 24 months. Check two sets of patients' VA and refraction before operation, and analyze the best-corrected visual acuity (BCVA), corneal refraction changes, corneal endothelium counting and complications, using χ(2) test and t-test. RESULTS: The BCVA after the operation of two sets are both improved, 32 eyes of DLK set were above 0.4, 19 eyes of PK set were above 0.4, of which the DKL set is a bit better than PK set (χ(2) = 4.304, P > 0.05). The astigmatism of DLK set is smaller than the PK set after operation, and there is significant difference (χ(2) = 4.98, P < 0.05). The astigmatism of two sets on the average were all no more than 5.00 D. The reject reaction of keratoplasty of DLK is less than PK, there is obviously significant difference (χ(2) = 34.17, P < 0.05). CONCLUSIONS: There is less complications of DLK than PK for fungal corneal ulcer. DLK can reduce the occurrence of reject reaction of endothelium type, and the BCVA, refraction of DLK after operation is similar to PK, the incidence rate of surgical operation failure is low.

Axial myopia induced by hyperopic defocus in guinea pigs: A detailed assessment on susceptibility and recovery.

This study investigated whether adolescent guinea pigs can develop myopia induced by negative lenses, and whether they can recover from the induced myopia. Forty-nine pigmented guinea pigs (age of 3 weeks) were randomly assigned to 4 groups: 2-week defocus (n=16), 4-week defocus (n=9), 2-week control (n=15) and 4-week control (n=9). A -4.00D lens was worn in the defocus groups and a plano lens worn in the control groups monocularly. The lenses were worn from 3 weeks to 5 weeks of age in the 2-week treatment groups with the biometry measured at 2, 4, 6, 10 and 14 days of lens wear. The lenses were worn from 3 weeks to 7 weeks of age in the 4-week treatment groups with the biometry measured immediately and at 2, 4, 6, 10 and 14 days after lens removal. Refractions in the defocused eyes developed towards myopia rapidly within 2 days of lens wear, followed by a slower development. The defocused eyes were at least 3.00D more myopic with a greater increase in vitreous length by 0.08 mm compared to the fellow eyes at 14 days (p<0.05). The estimated choroidal thickness of the defocused eyes decreased rapidly within 2 days of lens wear, followed by a slower decrease over the next 4 days. Relative myopia induced by 4 weeks of negative-lens treatment declined rapidly following lens removal. A complete recovery occurred 14 days after lens removal when compared to the fellow controls. The refractive changes during the recovery corresponded to a slower vitreous lengthening and a rapid thickening of the choroid. The plano-lens wearing eyes showed a slight but significant myopic shift (<-0.80D) with no associated biometrical changes. Guinea pigs aged 3 weeks can still develop negative lens induced myopia and this myopia is reversible after removal of the lens. The myopia and recovery are mainly due to changes in vitreous length and choroidal thickness.

A novel CRX mutation by whole-exome sequencing in an autosomal dominant cone-rod dystrophy pedigree.

AIM: To identify the disease-causing gene mutation in a Chinese pedigree with autosomal dominant cone-rod dystrophy (adCORD). METHODS: A southern Chinese adCORD pedigree including 9 affected individuals was studied. Whole-exome sequencing (WES), coupling the Agilent whole-exome capture system to the Illumina HiSeq 2000 DNA sequencing platform was used to search the specific gene mutation in 3 affected family members and 1 unaffected member. After a suggested variant was found through the data analysis, the putative mutation was validated by Sanger DNA sequencing of samples from all available family members. RESULTS: The results of both WES and Sanger sequencing revealed a novel nonsense mutation c.C766T (p.Q256X) within exon 5 of CRX gene which was pathogenic for adCORD in this family. The mutation could affect photoreceptor-specific gene expression with a dominant-negative effect and resulted in loss of the OTX tail, thus the mutant protein occupies the CRX-binding site in target promoters without establishing an interaction and, consequently, may block transactivation. CONCLUSION: All modes of Mendelian inheritance in CORD have been observed, and genetic heterogeneity is a hallmark of CORD. Therefore, conventional genetic diagnosis of CORD would be time-consuming and labor-intensive. Our study indicated the robustness and cost-effectiveness of WES in the genetic diagnosis of CORD.