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 evaluate the clinical effects of segmentectomy versus lobectomy under single utility port video-assisted thoracic surgery on inflammatory factors and immune cells in peripheral blood of non-small cell lung cancer patients, and to analyze the effect of changes of postoperative inflammatory factors and immune cells on the prognosis of the patients. Methods    The clinical data of 256 patients who underwent segmentectomy or lobectomy under single utility port video-assisted thoracic surgery for non-small cell lung cancer in the First Affiliated Hospital of Hebei North University from January 2016 to October 2020 were retrospectively collected. According to the operation method, they were divided into a segmentectomy group (126 patients with 79 males and 47 females at an age of 63.4±6.2 years) and a lobectomy group (130 patients with 91 males and 39 females at an age of 62.9±5.6 years). The change of inflammatory factors (C reactive protein, interleukin-6, interleukin-8, tumor necrosis factor-α) and immune cells (CD4+T cells, CD8+T cells and natural killer cells) were recorded and analyzed before operation (T0) and 1 day (T1), 3 days (T2), 7 days (T3), 1 month (T4) after the operation between the two groups. According to postoperative recurrence situations, they were divided into a recurrence group and a non-recurrence group, multivariate logistic regression analysis was used to analyze the relationship between the change of postoperative inflammatory factors, immune cells, and the prognosis of patients with non-small cell lung cancer. Results    (1) There was no statistical difference in sex ratio, underlying diseases, body mass index, levels of preoperative inflammatory factors or immune cells between the two groups (all P>0.05). (2) The changes of postoperative inflammatory factors in the segmentectomy group were significantly less than those in the lobectomy group at T1-T3 (all P<0.05), and the changes of postoperative immune cells in the segmentectomy group were significantly less than those in the lobectomy group at T1-T4 (all P<0.05). (3) The changes of postoperative inflammatory factors and immune cells on postoperative day 3 in the recurrence group were significantly more than those in the non-recurrence group (all P<0.05). (4) Multivariate logistic regression analysis showed that the changes of postoperative inflammatory factors and immune cells on postoperative day 3 may be the risk factors for postoperative recurrence and metastasis in patients with non-small cell lung cancer (all P<0.05). Conclusion    Single utility port video-assisted thoracic surgery segmentectomy for the treatment of non-small cell lung cancer can reduce the inflammatory response and protect body's immune function, and the change of postoperative inflammatory factors and immune cells in postoperative day 3 may be the risk factors for postoperative recurrence and metastasis in patients with non-small cell lung cancer.

Objective To evaluate the effectiveness of non-operative treatment for the acute intra-synovial sheath anterior cruciate ligament (ACL) rupture.Methods Twenty-eight patients diagnosed as the acute intra-synovial sheath ACL rupture at outpatient clinic between May 2014 and July 2016 were included.All patients were immobilized with knee braces for 6 weeks,followed by range of motion (ROM) training and partial to full weight-bearing of knees.All patients returned 3 months later for MRI scanning and those with the side-to-side difference of the anterior-posterior laxity less than 5 mm continued with non-operative treatment,followed up for MRI examination and clinical assessments 6 and 12 months later.Results Four patients dropped out because they didn't meet the stability criteria at 3 months after the treatment,3 of whom received surgical reconstruction and 1 with muscle strengthening training.Another patient received surgical reconstruction at 5 months due to re-injury.The remaining 23 patients achieved satisfactory results at 12 months after the treatment,with the average side-to-side difference of the anterior-posterior laxity of 2.1mm (0-4 mm),MRI good-to-excellent rate of 85.2％ (8 of Grade 1 and 15 of Grade 2),subjective IKDC (International Knee Documentation Committee) score of 92.71 (89.7-98.9),Lysholm score of 91.6 (86-95),and modified Larson score 96.4 (92-99).Conclusions Patients with the acute intra-synovial sheath anterior cruciate ligament (ACL) rupture showed satisfactory functional scores and objective stability and healing on MRI after the non-operative treatment.

Objective To evaluate the role of conventional protein kinase Cγ (cPKCγ)/growthassociated protein-43 (GAP-43) signaling pathway in ketamine-induced apoptosis in hippocampal neurons of developing rats in an in vitro experiment.Methods Primarily cultured hippocampal neurons were seeded in culture plates at a density of 1×10.6 cells/ml and divided into 2 groups (n=10 each) using a random number table:control group (C group) and ketamine group (K group).Group C received no treatment.Ketamine was added with the final concentration of 300 μmol/L in group K.At 12 h of culture or incubation,the apoptosis in hippocampal neurons was detected by flow cytometry.The apoptotic rate was calculated.The expression of cPKCγ,GAP-43 and phosphorylated GAP-43 in hippocampal neurons was measured by Western blot.Results Compared with group C,the apoptotic rates of hippocampal neurons were significantly increased,and the expression of cPKCγ,GAP-43 and phosphorylated GAP-43 was down-regulated in group K (P<0.01).Conclusion The mechanism by which ketamine induces apoptosis in hippocampal neurons of developing rats may be related to inhibition of cPKCγ/GAP-43 signaling pathway activation in an in vitro experiment.

OBJECTIVE:To compare anesthesia effects of 3 anesthesia methods,to investigate anesthesia improvement method for bronch fiber oscope. METHODS:315 patients undergoing bronch fiber oscope were randomly divided into group A(106 cas-es),group B(104 cases)and group C(105 cases). Group A was treated with traditional bronch fiber oscope lidocaine atomization local anesthesia,group B was treated with modified bronch fiber oscope pethidine combined with lidocaine atomization local anes-thesia and group C was treated with three-limb tube connected with laryngeal mask and target-controlled propofol and remifentanil general anesthesia. SaO2,SBP,DBP,heart rate,breathing rate,excellent anesthesia rate,compliance rate,the rate of bodymove-ment and choking,pain recalling rate were compared among 3 groups before and during examination,and ADR was observed. RE-SULTS:Compared with before examination,SaO2 of group A was decreased significantly during examination,with statistical sig-nificance(P<0.05);that of group B and C had no obvious change before and during examination. Blood pressure,heart rate and breathing rate of 3 groups had no obvious change before and during examination. The excellent anesthesia rate and compliance rate of group A were significantly lower than those of group B and C,but the rate of bodymovement cough and pain recalling rate were significantly higher than group B and C,with statistical significantly(P<0.05). The excellent anesthesia rate and compliance rate of group B were significantly lower than those of group C,but the rate of bodymovement cough and pain recalling rate were signifi-cantly higher than group C,with statistical significance (P<0.05). No ADR was found in 3 groups. CONCLUSIONS:Modified bronchoscope atomization local anesthesia and modified painless bronchoscope are better than traditional bronch fiber oscope atomi-zation local anesthesia in anesthesia effect,safety,degree of comfort and acceptability. Modified bronchoscope atomization local an-esthesia was best but most expensive,so these methods can be chosen according to patient’s condition.

Objective To evaluate the effect of age on the median-effective target plasma concentration (EC50) of propofol inhibiting body movement evoked by gastroscopy in the patients.Methods Ninety adult patients of both sexes,of ASA physical status Ⅰ or Ⅱ,with body mass index 19-25 kg/m2,scheduled for elective gastroscopy,were divided into 3 groups according to age (n =30 each):18-39 yr group (Ⅰ group),40-64 yr group (Ⅱ group) and 65-85 yr group (Ⅲ group).In Ⅰ,Ⅱ,Ⅲ groups,propofol was given by target-controlled infusion with the initial target concentrations of 2.5,2.0 and 1.5 μg/ml,respectively,and gastroscopy was performed when the target concentration was achieved.Body movement was defined as the directional movement in head or four extremities during gastroscopy.The target plasma concentration of propofol was determined by up-and-down sequential trial.Each time the plasma concentration of propofol increased/decreased by 0.5 μg/ml in the next patient depending on whether or not body movement developed.The EC50 and 95 ％ confidence interval of propofol inhibiting gastroscopy-evoked body movement were determined using Probit analysis.Results The EC50 (95 ％ confidence interval) of propofol was 4.2(3.8-4.5),4.1(3.7-4.4) and 2.4(1.8-2.7) μg/ml in Ⅰ,Ⅱ and Ⅲ groups,respectively.There was no significant difference in the EC50 of propofol between group Ⅱ and group Ⅰ.The EC50 of propofol was significantly lower in group Ⅲ than in Ⅰ and Ⅱ groups.Conclusion Age affects propofol-induced analgesia in patients with visceral pain,and the potency of propofol inhibiting visceral pain during gastroscopy in the elderly patients is significantly enhanced as compared with that in the young and middle-aged patients.

Objective To evaluate the effects of different doses of fentanyl on the median effective target plasma concentration (EC50) of propofol inhibiting body movement evoked by gastroscopy in the elderly patients.Methods Ninety patients of both sexes,aged 75-89 yr,with a body mass index of 19-25 kg/m2,of ASA physical status Ⅱ or Ⅲ,scheduled for elective gastroscopy,were randomly divided into 3 groups (n =30 each):control group (group C) and different doses of fentanyl groups (F0.5 and F1.0 groups).Fentanyl 0.5 and 1.0 μg/kg were injected intravenously in F0.5 and F1.0 groups,respectively.Propofol was then administered by target-controlled infusion.The initial target plasma concentrations (Cps) of propofol were 2.0,1.5 and 1.0 μg/ml in C,F0.5 and F1.0 groups,respectively.Gastroscopy was performed after the target effect-site and plasma concentrations were balanced.Body movement was defined as movement in head or four extremities during gastroscopy.The target Cp of propofol was determined by up-and-down sequential trial.Each time the Cp increased/decreased by 0.5 μg/ml in the next patient depending on whether or not body movement developed.The EC50 and 95 ％ confidence interval (CI) of propofol inhibiting gastroscopy-evoked body movement were determined using Probit analysis.Results The EC50 (95 ％ CI) of propofol was 2.24 ng/ml (1.67-2.47 ng/ml) in group C,1.79 (1.55-1.95) μg/ml in group F0.5,and 1.13 (1.08-1.62) μg/ml in group F1.0.There was no significant difference in the EC50 of propofol between F0.5 and C groups.The EC50 of propofol was significantly lower in F1.0 group than in C and F0.5 groups.Conclusion When combined with propofol,fentanyl 1.0 μg/kg is recommended for gastroscopy in the elderly patients.

The present paper is aimed to explore the biological osteoinductive activity of recombinant human bone morphogenetic protein 9 (rhBMP-9) by various biological technologies. In this study, we firstly obtained hBMP-9 cDNA by PCR and inserted it into vector pcDNA4/His Max to reconstruct hBMP-9 eukaryotic expression vector pcDNA4/His Max-BMP-9. Recombinant Chinese hamster ovary (rCHO) cell line expressing high-level rhBMP-9 was reconstructed by co-transfecting the expression vectors pcDNA4/His* Max-hBMP-9 and plasmid pSV2-dhfr into dihydrofolate reductase (dhfr)-deficient CHO cells and the subsequent gene amplification by the methotrexate. We finally obtained a monoclonal cell line expressing the highest level protein. We purified the medium after culturing the highest-producing monoclonal by Ni-NTA His-Bind Resin columns and concentrated to by a Centricon 50 at 4 degrees C and stored at 70 degrees C until it was used. Western blot and SDS-PAGE analyses showed a specific band of about 32kD in pro-region lane and a specific band of about 50kD in pro-region complex lane. Biological activities of rhBMP-9 were tested by colorimetric determination and histochemical staining of Alkaline Phosphatase (ALP) Activity, osteocalcin and oesteopontin for C3H10 T1/2 cells, which were stimulated culture by different concentration (20, 50, 100 microg/mL) of rhBMP-9. The results showed that the rhBMP-9 could induce osteogenic differentiation of mesenchymal stem cells (MSCs) in vitro, and were proportional to the amount. This study can provide experimental data for further tests in vivo and clinical applications.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Growth Differentiation Factor 2 ; biosynthesis ; isolation & purification ; pharmacology ; Humans ; Osteogenesis ; drug effects ; Recombinant Proteins ; biosynthesis ; isolation & purification ; pharmacology ; Transfection

A total of 6004 cases of acute poisoning with 532 kinds of related poisons from 63hospitals were analyzed.According to the classification of pharmaceutical,pesticide,chemical,animal,plant and other poisons,the numbers of poison categories were 217,148,61,34,36 and 36 kinds accounting for 40.8％,27.8％,11.5％,6.4％,6.8％,6.7％ of total number of poisons respectively.According to the case count,they were divided into three groups of low,medium and high morbidity and their poison numbers were 462,59 and 11 kinds accounting for 86.8％,11.1％,2.1％ of total number of poisons respectively.According to the types of poisons,they were divided into five groups of poisoning 1,2,3,4,5 years and their poison numbers were 320,91,34,33 and 54 kinds accounting for 60.2％,17.1％,6.4％,6.2％,10.1％ of total number of poisons respectively.According to the time of poisoning,they were divided into three groups of 1-year,discontinuous and perennial poisoning and their poison numbers were 320,158 and 54 kinds accounting for 60.2％,29.7％,10.1％ of total number of poisons respectively.

Objective To evaluate the efficacy of combination solifenacin and tamsulosin for the treatment of distal ureteral calculi after extracorporeal shock wave lithotripsy. Methods 120 patients (male:100 female:20 age:18-67 yrs) randomly assigned to 4 groups (each group 30) with the calculi diameter range from 0.5 to 1.1 cm.All patients performed extracorporeal shock wave lithotripsy (X ray oriented).The control group did not accept any medical treatment.The solifenacin group were administered solifenacin 5 mg,once per day.The tamsulosin group were administered tamsulosin 0.2 mg,once per day.The combination group were administered solifenacin 5mg,plus tamsulosin 0.2 mg,each per day.The observation duration was set at 2 weeks. Results The stone-free rate (according to KUB) within 2 weeks were 80.0％,83.3％,93.3％ and 96.7％ in the control group,solifenacin group,tamsulosin group and combination group respectively.Statistical differences were significant among the tamsulosin group,the combination group and the control group.The stone expulsion times were (7.6 + 3.7) d,(6.3 ± 2.5) d,(4.4 + 2.3) d and (3.5 ± 2.2) d in the 4 groups respectively.Statistical differences were significant among the tamsulosin group,the combination group and the control group.The uses of analgesics were 13,5,9 and 3 in the 4 groups respectively.The bladder irrtative symptoms were 12,6,4 and 4 in the 4 groups respectively.Statistical differences were also significant for the use of analgesics and relief of bladder irritation between the solifenacin group,the combination group and the control group. Conclusions Tamsulosin and solifenacin could be safe and effective for the treatment of distal ureteral calculi after extracorporeal shock wave lithotripsy.It could significantly improve the stone expulsion rate,relief the pain and improve bladder irrtative symptoms.