The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (M^pro) and papain like protease (PL^pro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851M^pro inhibitors and 205 PL^pro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both M^pro and PL^pro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of M^pro inhibitors and over 20% of PL^pro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 M^pro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.
Humans ; Antiviral Agents/chemistry* ; COVID-19 ; COVID-19 Drug Treatment ; High-Throughput Screening Assays ; Molecular Docking Simulation ; Protease Inhibitors/chemistry* ; SARS-CoV-2/enzymology* ; Viral Nonstructural Proteins

Objective:To explore the value of diffusion tensor imaging (DTI) in evaluating the effects of hyperbaric oxygen therapy (HOT) in treating spinal cord injury.Methods:The modified Allen′s method was used to induce a traumatic spinal cord injury in 30 rats who were then divided randomly into an injured group and a treatment group, each of 15. The treatment group was given HOT twice a day for 3 days, then once a day for a total of 4 weeks. The injured group did not receive HOT. DTI was performed (along with Basso-Beattie-Bresnahan (BBB) evaluation) at 0h, 6h, 24h, as well as 3, 7, 14, 21 and 28 days after the operation. Two-factor repeated measures ANOVA was conducted to analyze any differences in the DTI results: the fractional anisotropy, mean apparent diffusivity, radial diffusivity and axial diffusivity, as well as the BBB scores. LSD t-tests were performed to analyze the significance of the differences at different time points.Results:At each time point after 24h the average FA value of the treatment group was significantly higher than the injured group′s average, while its average MD and RD values were significantly lower. Beyond 14 days the average AD value of the treatment group was significantly higher than that of the injured group. The treatment group′s average BBB score was also significantly higher at all the time points beyond 3 days.Conclusions:DTI results can evaluate spinal cord function and provide valuable information for the dynamic assessment of hyperbaric oxygen therapy after a traumatic spinal cord injury, and the therapy promotes the recovery of motor function, at least in rats.

The exacerbation of progressive multiple sclerosis (MS) is closely associated with obstruction of the differentiation of oligodendrocyte progenitor cells (OPCs). To discover novel therapeutic compounds for enhancing remyelination by endogenous OPCs, we screened for myelin basic protein expression using cultured rat OPCs and a library of small-molecule compounds. One of the most effective drugs was pinocembrin, which remarkably promoted OPC differentiation and maturation without affecting cell proliferation and survival. Based on these in vitro effects, we further assessed the therapeutic effects of pinocembrin in animal models of demyelinating diseases. We demonstrated that pinocembrin significantly ameliorated the progression of experimental autoimmune encephalomyelitis (EAE) and enhanced the repair of demyelination in lysolectin-induced lesions. Further studies indicated that pinocembrin increased the phosphorylation level of mammalian target of rapamycin (mTOR). Taken together, our results demonstrated that pinocembrin promotes OPC differentiation and remyelination through the phosphorylated mTOR pathway, and suggest a novel therapeutic prospect for this natural flavonoid product in treating demyelinating diseases.

The exacerbation of progressive multiple sclerosis (MS) is closely associated with obstruction of the differentiation of oligodendrocyte progenitor cells (OPCs). To discover novel therapeutic compounds for enhancing remyelination by endogenous OPCs, we screened for myelin basic protein expression using cultured rat OPCs and a library of small-molecule compounds. One of the most effective drugs was pinocembrin, which remarkably promoted OPC differentiation and maturation without affecting cell proliferation and survival. Based on these in vitro effects, we further assessed the therapeutic effects of pinocembrin in animal models of demyelinating diseases. We demonstrated that pinocembrin significantly ameliorated the progression of experimental autoimmune encephalomyelitis (EAE) and enhanced the repair of demyelination in lysolectin-induced lesions. Further studies indicated that pinocembrin increased the phosphorylation level of mammalian target of rapamycin (mTOR). Taken together, our results demonstrated that pinocembrin promotes OPC differentiation and remyelination through the phosphorylated mTOR pathway, and suggest a novel therapeutic prospect for this natural flavonoid product in treating demyelinating diseases.
Animals ; Cell Differentiation ; Flavanones ; Mice ; Mice, Inbred C57BL ; Myelin Sheath/metabolism* ; Oligodendroglia/metabolism* ; Rats ; Remyelination ; Signal Transduction ; TOR Serine-Threonine Kinases/metabolism*

Objective: The purpose of our study was to investigate the predictive abilities of clinical and computed tomography (CT)features for outcome prediction in patients with coronavirus disease (COVID-19). Materials and Methods: The clinical and CT data of 238 patients with laboratory-confirmed COVID-19 in our two hospitalswere retrospectively analyzed. One hundred sixty-six patients (103 males; age 43.8 ± 12.3 years) were allocated in thetraining cohort and 72 patients (38 males; age 45.1 ± 15.8 years) from another independent hospital were assigned in thevalidation cohort. The primary composite endpoint was admission to an intensive care unit, use of mechanical ventilation, ordeath. Univariate and multivariate Cox proportional hazard analyses were performed to identify independent predictors. Anomogram was constructed based on the combination of clinical and CT features, and its prognostic performance wasexternally tested in the validation group. The predictive value of the combined model was compared with models built on theclinical and radiological attributes alone. Results: Overall, 35 infected patients (21.1%) in the training cohort and 10 patients (13.9%) in the validation cohortexperienced adverse outcomes. Underlying comorbidity (hazard ratio [HR], 3.35; 95% confidence interval [CI], 1.67–6.71;p < 0.001), lymphocyte count (HR, 0.12; 95% CI, 0.04–0.38; p < 0.001) and crazy-paving sign (HR, 2.15; 95% CI, 1.03–4.48;p = 0.042) were the independent factors. The nomogram displayed a concordance index (C-index) of 0.82 (95% CI, 0.76–0.88),and its prognostic value was confirmed in the validation cohort with a C-index of 0.89 (95% CI, 0.82–0.96). The combinedmodel provided the best performance over the clinical or radiological model (p < 0.050). Conclusion Underlying comorbidity, lymphocyte count and crazy-paving sign were independent predictors of adverseoutcomes. The prognostic nomogram based on the combination of clinical and CT features could be a useful tool for predictingadverse outcomes of patients with COVID-19.

The differentiation and maturation of oligodendrocyte precursor cells (OPCs) is essential for myelination and remyelination in the CNS. The failure of OPCs to achieve terminal differentiation in demyelinating lesions often results in unsuccessful remyelination in a variety of human demyelinating diseases. However, the molecular mechanisms controlling OPC differentiation under pathological conditions remain largely unknown. Myt1L (myelin transcription factor 1-like), mainly expressed in neurons, has been associated with intellectual disability, schizophrenia, and depression. In the present study, we found that Myt1L was expressed in oligodendrocyte lineage cells during myelination and remyelination. The expression level of Myt1L in neuron/glia antigen 2-positive (NG2) OPCs was significantly higher than that in mature CC1 oligodendrocytes. In primary cultured OPCs, overexpression of Myt1L promoted, while knockdown inhibited OPC differentiation. Moreover, Myt1L was potently involved in promoting remyelination after lysolecithin-induced demyelination in vivo. ChIP assays showed that Myt1L bound to the promoter of Olig1 and transcriptionally regulated Olig1 expression. Taken together, our findings demonstrate that Myt1L is an essential regulator of OPC differentiation, thereby supporting Myt1L as a potential therapeutic target for demyelinating diseases.
Animals ; Cell Differentiation ; physiology ; Demyelinating Diseases ; chemically induced ; Lysophosphatidylcholines ; toxicity ; Mice ; Mice, Inbred C57BL ; Nerve Tissue Proteins ; metabolism ; Oligodendrocyte Precursor Cells ; cytology ; metabolism ; Oligodendroglia ; cytology ; metabolism ; Remyelination ; physiology ; Transcription Factors ; metabolism

Objective To investigate the change features of the 3-dimensional speckle tracking imaging (3D-STI) for quantitative evaluation of whole strain of left ventricular before and after left atrial appendage closure in the patients with non valvular atrial fibrillation and to evaluate its clinical application value.Methods Thirty patients who received left atrial appendage (LAA) closure from June, 2016 to A-pril, 2017 in Changhai Hospital were enrolled as research subjects .The patients received routine ultrasonic cardiography and 3D-STI, and left ventricular fractional shortening (LVFS), left ventricular ejection fractions (LVEF), global peak longitudinal strain (GPLS), global peak circumferential strain (GPCS), global peak radial strain (GPRS)as well as global peak area strain (GPAS) were respectively detected before surgery, 2 days, 1 and 3 months after surgery.Results In the 30 patients who received LAA closure, LVFS and LVEF were evaluated by u-sing routine ultrasonic cardiography and the data detected before surgery , 2 days, 1 and 3 months after surgery were compared between the two, and results showed that there was no statistical significance, when comparisons were made between them (P>0.05).In the 30 patients who were implemented with LAA closure, GPLS, GPCS, GPRS and GPAS values were detected using 3D-STI.The detected data 2 days after surgery were significantly higher as compared with those before surgery and 1 and 3 months after surgery(e.g.GPAS:39.90 ±10.22/27.80 ± 5.00, 26.20 ±6.05, 27.60 ±7.58), and there was statistical significance, when comparisons were made between them (P<0.05).However, there was no statistical significance in the data of GPLS, GPRS, GPCS and GPAS detected before surgery and 1 and 3 months after surgery, when comparisons were made between the two (P>0.05).Conclusion 3D-STI could be used for quantitative evaluation of the changes in left ventricular systolic function before and after LAA closure.The whole strain of the left ventricle of the patient was considerably increased 2 days after surgery, and the whole strain of the left ventricle recovered to pre-surgical level one month after surgery.

Objective·To compare the clinical features between different subtype bipolar patients with first mania episode, and to contribute to early identification of bipolar disorder. Methods·This study was based on the database named as National Bipolar Mania Pathway Survey (BIPAS). From November 2012 to January 2013, bipolar patients from 26 mental health facilities in China were enrolled in current study. The clinical features were compared between mania patients of different subtypes, including hypomania (groupⅠ), mania without psychotic symptoms (groupⅡ), mania with psychotic symptoms (group Ⅲ) and mixed state (group Ⅳ). Results·There was significant difference in the percentage of clinical symptoms between different subtype bipolar patients with first mania episode, especially the mania and anxiety related symptoms. Group Ⅰ, Ⅲ , Ⅳ were further compared with groupⅡ, which was considered as the typical bipolar disorder. The results showed that the mania related symptoms was significantly higher in group Ⅱ, but anxiety related symptoms was significantly higher in group Ⅰ, Ⅲ, Ⅳ. Moreover, Logistic regression analysis revealed that more eloquent or humor and unusually restless could be in favor of the diagnosis of hypomania; younger and mania or hypomania as first episode might be in favor of the diagnosis of mania with psychotic symptoms; older, national minorities and unusually restless could be in favor of the diagnosis of mixed state. Conclusion·The clinical features between different subtype bipolar patients with first mania episode are various, and analysis of the clinical features can contribute to early identification of bipolar disorder.

Objective: To explore the application of real-time three-dimensional ultrasound in locating the course, the brand and the blind end of internal opening of congenital preauricular fistula. Methods: Congenital preauricular fistula patients from Changhai Hospital were studied. All the cases were evaluated using two-dimensional and three-dimensional ultrasound before surgeries. The surgery findings were utilized as the gold standard. The imaging features of 3D ultrasound were described to determine the accuracy of the blind end location, the course of the main fistula tract and the branch tracts. The diagnosis was compared with the surgery findings. Results: In 89 cases, among which there were 21 branch fistulae. There were 68 bend fistulae without branch. The diagnostic accuracy of both two-dimensional ultrasound and three-dimensional ultrasound was 100%. The accuracy in locating the blind end of internal opening, the course of the branch tracts with 3D ultrasound was 92.1%(82/89), 85.7%(18/21) , compared to 80.9%(72/89), 57.1%(12/21) with 2D ultrasound. It was obvious that 3D ultrasound had more advantages. These differences were statistically significant (χ²=4.8, P<0.05; χ²=4.2, P<0.05). Conclusions Compared with 2D ultrasound, 3D ultrasound can accurately locate the blind end of internal opening, the course, and the branch of preauricular fistula, clearly display the morphology and the spatial structure of preauricular fistulaand its surroundings. It has high clinical application value in preoperative imaging examination.

Objective To observe the morphology changes in articular cartilage of knee osteoarthritis ( KOA) model rats after intervention with Shudi Jisheng Zhuanggu Decoction (SJZD). Methods SD rats were randomized into 7 groups, namely blank control group, model group, Paracetamol group ( 0.270 g/kg) , Zhuangu Guanjie Pills group ( 1.08 g/kg) , and high-, middle- and low-dosage SJZD groups ( 27.450, 13.725, 6.863 g/kg, respectively). Rats were injected with papain solution into the right knee joint to build models of KOA before administration of drugs. Thickness of right knee joint was measured to evaluate the effect of SJZD on the swelling degree of knee joint. Histological changes of knee cartilage were evaluated by Mankin scoring method after the cartilage sections being stained with hematoxylin-eosin (HE). Results Swelling degree of knee joint and Mankin scores were significantly increased in the model group compared to the blank control group ( P<0.01). In comparison with the model group, the swelling degree of knee joint and Mankin scores of SJZD groups were significantly decreased (P<0.05 or P<0.01), and the pathological changes were relieved. Conclusion SJZD can ameliorate the pathological changes of knee articular cartilage in rats with KOA, so as to delay cartilage degeneration and protect the cartilage.