The emergence of SARS-CoV-2 variants of concern and repeated outbreaks of coronavirus epidemics in the past two decades emphasize the need for next-generation pan-coronaviral therapeutics. Drugging the multi-functional papain-like protease (PLpro) domain of the viral nsp3 holds promise. However, none of the known coronavirus PLpro inhibitors has been shown to be in vivo active. Herein, we screened a structurally diverse library of 50,080 compounds for potential coronavirus PLpro inhibitors and identified a noncovalent lead inhibitor F0213 that has broad-spectrum anti-coronaviral activity, including against the Sarbecoviruses (SARS-CoV-1 and SARS-CoV-2), Merbecovirus (MERS-CoV), as well as the Alphacoronavirus (hCoV-229E and hCoV-OC43). Importantly, F0213 confers protection in both SARS-CoV-2-infected hamsters and MERS-CoV-infected human DPP4-knockin mice. F0213 possesses a dual therapeutic functionality that suppresses coronavirus replication via blocking viral polyprotein cleavage, as well as promoting antiviral immunity by antagonizing the PLpro deubiquitinase activity. Despite the significant difference of substrate recognition, mode of inhibition studies suggest that F0213 is a competitive inhibitor against SARS2-PLpro via binding with the 157K amino acid residue, whereas an allosteric inhibitor of MERS-PLpro interacting with its 271E position. Our proof-of-concept findings demonstrated that PLpro is a valid target for the development of broad-spectrum anti-coronavirus agents. The orally administered F0213 may serve as a promising lead compound for combating the ongoing COVID-19 pandemic and future coronavirus outbreaks.
Animals ; Coronavirus Papain-Like Proteases/antagonists & inhibitors* ; Cricetinae ; Humans ; Mice ; Pandemics ; SARS-CoV-2/enzymology* ; COVID-19 Drug Treatment

Objective To explore the incidence and risk factors of perioperative major adverse cardiac events (MACE) in lung cancer patients with high-risk coronary heart disease undergoing pulmonary lobectomy. Methods The clinical dataof 1 647 high-risk coronary heart disease patients diagnosed with lung cancer undergoing lobectomy in our hospital was analyzed, and performed Framingham scoring. High-risk patients (score＞ 20％) were included in the study, and the periopertive major adverse cardiac events was defined as primary endpoint. The risk factors of MACE were analyzed. Results Perioperative MACE occurred in 26. 4％ of lung cancer patients with high-risk coronary heart disease undergoing lobectomy. Multivariate analysis demonstrated that hypertension, high density lipoprotein (HDL-C), diabetes, age, coronary angiography, stroke, smoking index in descending sequence were independent risk factors of perioperative cardiac events in lung cancer patients. While shorter operative time, coronary angiography and clinical intervention was protective factor. Conclusion Lung cancer patients with high-risk coronary heart disease undergoing lobectomy has higher risk of perioperative MACE. Preoperative sufficient cardiac risk scores, coronary angiography andclinical interventioncan reduce the incidence of perioperative MACE in lung cancer patients with high-risk coronary heart disease.

Objective To explore the incidence and risk factors of perioperative major adverse cardiac events (MACE) in lung cancer patients with high-risk coronary heart disease undergoing pulmonary lobectomy. Methods The clinical dataof 1 647 high-risk coronary heart disease patients diagnosed with lung cancer undergoing lobectomy in our hospital was analyzed, and performed Framingham scoring. High-risk patients (score＞ 20％) were included in the study, and the periopertive major adverse cardiac events was defined as primary endpoint. The risk factors of MACE were analyzed. Results Perioperative MACE occurred in 26. 4％ of lung cancer patients with high-risk coronary heart disease undergoing lobectomy. Multivariate analysis demonstrated that hypertension, high density lipoprotein (HDL-C), diabetes, age, coronary angiography, stroke, smoking index in descending sequence were independent risk factors of perioperative cardiac events in lung cancer patients. While shorter operative time, coronary angiography and clinical intervention was protective factor. Conclusion Lung cancer patients with high-risk coronary heart disease undergoing lobectomy has higher risk of perioperative MACE. Preoperative sufficient cardiac risk scores, coronary angiography andclinical interventioncan reduce the incidence of perioperative MACE in lung cancer patients with high-risk coronary heart disease.

This study was to develop a quantitative immunoassay for hyaluronic acid(HA). We firstly prepared a monoclonal antibody(MAb)against it. HA, activated by 1-cyano-4-dimethyl-aminopyridinium tetrafluoroborate(CDAP), was coupled to bovine serum albumin(BSA). BALB/c mice were immunized with purified complete antigen BSA-HA and the monoclonal antibody was prepared by hybridoma technique. To characterize the specificity of the antibody, inhibition enzyme-linked immunosorbent assay(ELISA)was conducted and the cross reactivity along with affinity constant was determined. The results showed that the substitution degree of BSA-HA(mBSA/wHA)was about 9. Also, a hybridoma cell line named 8B6 was obtained by hybridoma technology. The indirect ELISA titer of the ascetic fluid was 1 ∶256 000; the isotype of MAb 8B6 was IgG1 and the affinity constant was Ka=6. 71×109 mol-1. To establish the indirect competitive ELISA method, polystyrene microwell plates were coated with 10. 0 ng/mL HA and blocked with 1. 0% BSA. The linear range of indirect competitive ELISA was between 10. 0 ng/mL to 10. 0 μg/mL which confirmed that the method was specific and sensitive.

Objective:To construct NMB0315 eukaryotic expression recombinant vector ,detect specific humoral and cellular immune response induced by the recombint DNA vaccine intramuscularly in female BALB /c mice,evaluate the immunocompetence and immunoprotection of the vaccine , so as to provide experimental basis for the development of a novel nucleic acid vaccine against N.meningitidis serogroup B .Methods: The whole NMB0315 gene was amplified by PCR from the standard strains MC 58 genomic DNA,cloned into a plasmid pcDNA3.1(+),identified by double digestion of the recombinant plasmid with restriction enzymes and se -quencing.The recombinant vector pcDNA 3.1 (+)/NMB0315 was transfected into eukaryotic COS-7 cells and RAW264.7 cells, the NMB0315 protein was detected by immunocytochemical method and Western blot respectively .The levels of specific humoral and cellular immune response were detected after inoculating in female BALB /c mice intramuscularly with the recombinant plasmid .The immune protective effect was investigated with the DNA vaccine and the bactericidal titer of the immune serum was deter mined by serum bactericidal assay ( SBA ) in vitro.Results: The recombinant pcDNA3.1 (+)/NMB0315 was effectively transcripted and expressed in eukaryotic cells and the specific humoral and cellular immune responses were induced in the inoculated mice .In the re-combinant pcDNA3.1(+)/NMB0315 group ,the levels of serum IgG,IgG1,IgG2a,IgG2b and IgG3 and genital tract sIgA were significantly higher than in controls ( P<0.001 ) .The stimulation index in the culture supernatant of the spleen lymphocytes of the vaccine group was higher than that of the control group (P<0.05).The ratios of serum IgG2a/IgG1 in the DNA vaccine group were less than 1.The bactericidal titer of the NMB 0315+CpG group reached 1:128 following three immunizations , the protection rate of the vaccine group was 70%against the N.meningitidis strain MC58.Conclusion:The NMB0315 nucleic acid vaccine could induce higher levels of humoral immunity and cellular immunity and showed effective protection against N .meningitidis serogroup B , the immune serum had strong bactericidal activity in vitro .

Objective This retrospective study is to analysis the special medical conditions that most Chinese secondary hospitals are facing with,and to review the safeguards and pitfalls for arterial switch operations,in order to probe intothe feasibility of this procedure for Chinese secondary hospitals and provide our experiences to help other surgeons to avoid pitfalls on complex procedures.Methods Between January 2006 and December 2011,totally 21 newborns and infants with TGA/VSD and TGA/IVS underwent arterial switch operation.There were 15 males and 6 females.In the TGA/VSD group,there were 16 cases,ranging from 30 days to 1 year at surgeries,and weight from 3.4-8.5 kg with average of (5.33 ± 1.42) kg.In the TGA/IVS group,there were 5 cases,ranging from 13 days to 1 month at surgeries,and weight from 3.1-5.5 kg with average of (3.75 ± 1.17)kg.All patients underwent one stage of arterial switch operation.Routine follow-up checking points are set at discharging,3 months,half year and every year after operation.Results The early death rate is 9.5％ (2/21),and the reexploration rate is 9.5％ (2/21).In the TGA/VSD group,average cardiopulmonary bypass time is (151 ± 33) minuntes with the aortic crossclamp time is (1 19 ± 26) minutes.Ventilator support time is 24-159 hours,and the length of ICU stay is 3-17 days.1 case has residual VSD with the diameter less than 2 mm.The pulmonary flow velocity in 2 cases increase mildly to 2.0 m/s and 2.2 m/s,and another 2 cases increase severely to 3.1 m/s and 3.7 m/s.The aortic flow velocity in 3 cases increase to 2.0m/s.ECG instructs no case has myoinfarction signs.In the TGA/IVS group,average cardiopulmonary bypass time is (170 ± 52) minuntes with the aortic crossclamp time is (137 ± 48) minutes.Ventilator support time is 51-144 hours,and the length of ICU stay is 4-14 days;The pulmonary flow velocity in 2 cases increase mildly to 2.0 m/s.The aortic flow velocity in 1 cases increase to 2.0 m/s.ECG instructs no case has myoinfarction signs.Conclusion Unbalanced medical resources distribution causes significant differences between heart centers and the secondary hospitals in China,especially on the complex congenital heart diseases procedures.However,with relatively solid background on correction of the simple congenital heart diseases,the Chinese secondary hospitals can still perform arterial switch operation with satisfactory mortality and morbidity,and provide more prompt medical services for more population.

ObjectiveTo investigate the effects of isoflurane anesthesia on hippocampus synaptosomes proteome in aged rats.MethodsTwenty-seven 22- month-old SD rats weighing 480-550 g were randomly divided into 2 groups: control group (group C,n =6) and isoflurane group (group Ⅰ,n =21 ).In group C inhaled mixed gas containing 80％ oxygen for 2 h.In group Ⅰ the animals were endotracheal intubated after induction by 3％ isoflurane and inhaled 2％ isoflurane and 80％ oxygen for 2 h.Cognition function was evaluated by Y-maze at 24 h after anesthesia and the total training times were recorded.The total training times ＞ 75 was defined as cognitive dysfuction.In group Ⅰ the animals were divided into cognitive dysfuction group (group ⅠA) and non-cognitive dysfuction group (group IB) according to the results of Y-maze test.The animals were sacrificed and their hippocampi were removed and synaptosomes were extracted for two-dimensional gel electrophoresis.The different protein spots were analyzed by mass chromatographic analysis.ResultsSix rats had cognitive dysfuction (group IA) and another thirteen rats had no cognitive dysfuction (group IB).The total training times were significantly higher in group IA than in groups C and IB( P ＜ 0.05).There was no significant difference in the total training times between groups C and IB (P ＞ 0.05).There were 21 (11/10) different protein spots between groups IB and IA,and 19 (12/7) different protein spots between groups C and IA.Thirty-one protein spots were identified by means of MALDI-TOF-MS.ConclusionThe cognitive dysfuction after isoflurane anesthesia in aged rats may be related to the changes of energy metabolism protein,cytoskeletal structure and regulatory protein in synapse of hippocampus.

ObjectiveTo investigate the effects of dexmedetomidine on postoperative cognitive function and monocytes Toll-like receptor 2 (TLR2)and TLR 4 expression in elderly patients.MethodsForty-five ASA Ⅰ or Ⅱ elderly patients aged ≥65 yr weighing 53-72 kg were randomly divided into 3 groups: control group (group Ⅰ ) and different doses of dexmedetomidine groups(groups Ⅱ and Ⅲ ).Dexmedetomidine 1.0 μg/kg was injected iv over 15 min after anesthesia induction,and then was infused at a rate of 0.5 μg·kg-1 ·h-1 (group Ⅱ ) or 1.0 μg· kg-1 ·h-1 (group Ⅲ ) untile the end of operation.Group Ⅰ received equal volume of normal saline.Blood samples were taken before anesthesia induction,at 1.5 h after the beginning of operation,at the end of operation and at 24 h after operation(T,-T5 ) for determination of monocytes TLR2 and TLR4 expression by flow cytometrybased method.Postoperative cognitive function was evaluated at 1 d before and 7 d after operation with Mini-mental state examination and Wechsler memory scale and Wechsler adult intelligence scale,and the postoperative cognitive dysfunction was recorded.ResultsThe incidence of postoperative cognitive dysfunction and monocytes TLR2 and TLR4 expression were significantly lower in groups Ⅱ and Ⅲ than in group Ⅰ,and in group Ⅲ than in group Ⅱ (P ＜ 0.05).ConclusionDexmedetomidine can prevent postoperative cognitive dysfunction in elderly patients,and the mechanism may be related to down-regulation of monocytes TLR2 and TLR4 expression.

ObjectiveTo investigate the effects of aliskiren on blood nitric oxide(NO) and bradykinin (BK) through analyzing the changes of NO, BK and non-dominant arm blood pressure before and after treatment of aliskiren and ramipril. MethodsThe chnical trial was conducted in 67 patients with essential hypertension. After a single-blind period of taking placebo orally once a day for 2 weeks, the patients were divided into different group in accordance with random table, and all patients were consecutively treated with drugs for 8 weeks. The trial uncovered showed that 17 patients were treated with ramipril (5 mg) in control group. Patients in trial group were given aliskiren and were assigned to three subgroups, 300 mg group (300 mg aliskiren, 16 cases), 150 mg group(150 mg aliskiren, 17 cases) and 75 mg group(75 mg aliskiren, 17 cases). The blood NO and BK before and after treatment in two groups were measured by enzyme linked immunosorbent assay and radioimmunoassay method. Non-dominant arm blood pressure was measured by calibration qualified mercury blood pressure instrument before and after treatment. Results The blood NO after treatment of aliskiren 8 weeks in trial group increased significantly than those before treatment [before treatment, the blood NO in 300 mg group, 150 mg group,75 mg group were (44.414 ±5.841 ), (43.496 ± 5.576), (41.037 ± 5.312) μ mol/L, after treatment they were(60.381 ± 6.756), (56.480 ±6.959), (53.766 ±7.276) μmol/L] (P <0.05). After treatment, non-dominant arm blood pressure decreased significantly in trial group (P < 0.05 ), but the blood BK had no significant defference before and after treatment (P >0.05). The blood NO and BK after treatment of ramipril 8 weeks in control group increased significantly than those before treatment [(57.286 ±6.431) μmol/L vs.(39.935 ±6.388)μ mol/L, (7.120 ± 1.015) μg/L vs.(5.232 ± 1.288) μg/L], and meanwhile non-dominant arm blood pressure decreased significantly(P <0.05). ConclusionsAliskiren and ramipril could increase the concentration of NO remarkably. Ramipril has strong effect in increasing the concentration of BK, but aliskiren hasn't effect on BK.

Objective To study the feasibility, safety and clinical effects of spleen and splenic vessel-preserving distal pancreatectomy. Methods A retrospective study was performed in 26 patients undergoing distal pancreatectomy for benign or low grade malignant disease with splenectomy (n = 13) or splenic preservation (n = 13 ) at the First Hospital of Sun Yat-sen University and Guangdong General Hospital from May 2002 to April 2009. Results All 26 pancreatectomy with splenectomy or splenic preservation were performed successfully. There was no statistically significant difference between two groups in average operative time[(172±47) min vs. (157±52) min, P ＞ 0.05 ], intraoperative estimated blood loss [( 183 ± 68 ) ml vs. ( 160 ± 51 ) ml, P ＞ 0.05 ], incidence of noninfectious and infection complication and postoperative hospital stay [(10.1±2.2) d vs. ( 12. 1 ± 4. 6 ) d, P ＞ 0.05 ]. The platelet counts examined one week after operation were significantly higher in the distal pancreatectomy with splenectomy group than that in spleen-preserving group [(37.3 ± 12.8)×109/L vs. (54.7 ± 13.2) × 109/L, P＜0.05 ]. Conclusions Spleen-preserving distal pancreatectomy appears to be a feasible and safe procedure in selected cases of benign or low-grade pancreatic malignant disease necessitating a distal pancreatectomy.