The coronavirus disease 2019 (COVID-19) is an acute infectious disease caused by the new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, which has led to serious worldwide economic burden. Due to the continuous emergence of variants, vaccines and monoclonal antibodies are only partial effective against infections caused by distinct strains of SARS-CoV-2. Therefore, it is still of great importance to call for the development of broad-spectrum and effective small molecule drugs to combat both current and future outbreaks triggered by SARS-CoV-2. Cathepsin L (CatL) cleaves the spike glycoprotein (S) of SARS-CoV-2, playing an indispensable role in enhancing virus entry into host cells. Therefore CatL is one of the ideal targets for the development of pan-coronavirus inhibitor-based drugs. In this study, a CatL enzyme inhibitor screening model was established based on fluorescein labeled substrate. Two CatL inhibitors IMB 6290 and IMB 8014 with low cytotoxicity were obtained through high-throughput screening, the half inhibition concentrations (IC₅₀) of which were 11.53 ± 0.68 and 1.56 ± 1.10 μmol·L^-1, respectively. SDS-PAGE and cell-cell fusion experiments confirmed that the compounds inhibited the hydrolysis of S protein by CatL in a concentration-dependent manner. Surface plasmon resonance (SPR) detection showed that both compounds exhibited moderate binding affinity with CatL. Molecular docking revealed the binding mode between the compound and the CatL active pocket. The pseudovirus experiment further confirmed the inhibitory effects of IMB 8014 on the S protein mediated entry process. In vitro pharmacokinetic evaluation indicated that the compounds had relatively good drug-likeness properties. Our research suggested that these two compounds have the potential to be further developed as antiviral drugs for COVID-19 treatment.

Objective To study experienced doctors’ treatment of senile dementia based on association rule. Methods Databases, such as CNKI, CSCD, CBMdisc, CPD, and Wanfang Data were searched for articles about senile dementia. SPSS17.0 and Clementine12.0 software were used for frequency and correlation analysis of prescription. Results In total of 67 articles about TCM therapy for senile dementia were included, containing 112 prescriptions, 178 Chinese herbal medicines, 1589 frequency, and 14.2 entries for each prescription. The first three frequently used herbal medicines were Acori Tatarinowii Rhizoma, Polygala Radix, and Salviae Miltiorrhizae Radix et Rhizoma. Accoring to association rule, the first three medicine pairs were Citrus Reticulatae Pericarpium→Pinelliae Rhizoma, Dioscoreae Rhizoma→Rehmanniae Radix Praeparata, and Codonopsis Radix→Citrus Reticulatae Pericarpium. The first three thriple herbal medicines were Dioscoreae Rhizoma+Lycii Fructus→Corni Fructus, Acori Tatarinowii Rhizoma+Corni Fructus→Rehmanniae Radix Praeparata, and Carthami Flos + Hirudo→Bupleuri Radix. Conclusion The results based on association rule effectively summarized all the doctors’ treatment experience in senile dementia, and provided the beneficial reference for the clinical treatment of senile dementia.

This paper summarize the practice in the teaching reform of microbiology experiment in recent years. We identify the main contents of experimental teaching systems and pay much more attention to peo-ple-oriented. Through the reform of teaching and assessment methods,students are trained to cultivate their practical ability and spirit of innovation.

Transmembrane serine protease 2 (TMPRSS2) is a cell surface protease widely present in the human body. It is involved in the infection of various viruses such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and in the cell invasion, tumor growth and metastasis processes of prostate cancer. This study used Boc-Gln-Ala-Arg-AMC as the fluorescent substrate to determine the cleavage activity of TMPRSS2 towards SARS-CoV-2 S protein. Then cell-based screening model for TMPRSS2 inhibitors was established in Vero E6 cells overexpressing TMPRSS2 (Vero E6/TMPRSS2). Seven compounds exhibiting TMPRSS2 inhibitory activities with low toxicity were obtained through high-throughput screening (HTS) from natural and synthetic compound pure product library of National Center for Screening Novel Microbial Drugs. Surface plasmon resonance (SPR) has shown that the obtained inhibitors could bind to TMPRSS2 with moderate affinity in a dose dependent manner. Cell-cell fusion experiments have shown that the obtained inhibitors can inhibit the occurrence of S protein mediated cell-cell fusion by inhibiting TMPRSS2 cleavage of SARS-CoV-2 S protein in a concentration dependent manner. Preliminary pseudovirus experiment showed that the inhibitors may reduce the pseudovirus infection into Opti-HEK-293T-ACE2 cells to varying degrees. In a word, this study successfully established a cell-based HTS model for TMPRSS2 inhibitor and preliminarily confirmed that the seven screened inhibitors possessed in vitro anti-TMPRSS2 activities, providing new structural scaffolds for the development of new drugs against SARS-CoV-2.

Objective To investigate the clinical effect of pulmonary rehabilitation therapy including respiratory exercise and vibration expectoration on patients with pulmonary infection after abdominal surgery.Methods A retrospective case control study was conducted.Seventy-six patients with pulmonary infection after abdominal surgery admitted to the First Affiliated Hospital of Hunan Normal University from September 2015 to September 2016 were enrolled.According to whether accept the pulmonary rehabilitation therapy or not,the patients were divided into two groups.In the control group (n =35),the convemional expectoration method was adopted.The patients in pulmonary rehabilitation group (n =41) received both methods of the control group and pulmonary rehabilitation treatment,including respiratory exercise (effective cough,lip reduction breathing),respiratory exercise device (respiratory exerciser tri-ball),and vibrated expectoration.The 24-hour sputum volume,degree of comfort,inflammatory and pulmonary function parameters,and recovery situation were recorded in the two groups.Results ① There were no significant differences in the parameters of inflammation and pulmonary function before treatment between the two groups.After treatment,the white blood cell (WBC) and C-reactive protein (CRP) in both groups were significantly decreased,and the forced expiratory volume in 1 second (FEV1) and FEV1/forced vital capacity (FVC) were significantly increased.The above changes in pulmonary rehabilitation group were more significant than those of the control group [WBC (× 109/L):11.12 ± 2.88 vs.13.42 ± 2.62 at 3 days,8.22 ± 1.48 vs.9.27 ± 1.92 at 5 days;CRP (mg/L):13.47 ± 4.77vs.16.03±4.94 at 3 days,9.69±1.56 vs.11.77±1.41 at 5 days;FEV1 (L):2.48±0.14 vs.2.29±0.16 at 3 days,FEV1/FVC:0.78±0.04 vs.0.75±0.04 at 3 days;all P ＜ 0.05].② The 24-hour sputum volume within 3 days of pulmonary rehabilitation group were significantly higher than that of the control group (mL:30.51 ± 4.15 vs.18.30 ± 3.64at 1 day,31.08±3.22 vs.20.37±3.20 at 2 days,29.03±2.55 vs.19.03±2.51 at 3 days,all P ＜ 0.01].③ In the pulmonary rehabilitation group,the recovery time of pulmonary infection symptoms (days:5.44 ± 1.45 vs.6.20 ± 1.55),the days of antibiotic use (days:12.61 ± 3.15 vs.15.03 ± 3.78),the time of getting out of the bed (days:4.05 ± 0.74vs.4.51±0.89),and the hospital days (days:19.95±3.90 vs.22.00±4.42) were significantly shorter than those of the control group (all P ＜ 0.05),and the degree of comfort was significantly better than that of the control group (comfort score:2.71 ±0.90 vs.2.14±0.91,P ＜ 0.01).Conclusion The application of pulmonary rehabilitation including respiratory exercise and vibration expectoration in abdominal surgery patients with pulmonary infection can promote recovery,and it has a good clinical and practical application value.

OBJECTIVEThis paper aims to develop a monoclonal antibodies (MAbs)- based ELISA for detecting Chlamydophila pneumoniae (C. pneumoniae) antigens in humans with the variable domains (VD) 2 and 3 of the major outer membrane protein (MOMPVD2-VD3) and to assess its sensitivity and specificity by comparing with a widely used MAb that is able to recognize the elementary bodies of C. pneumoniae.

METHODSMOMPVD2-VD3 were overexpressed in Escherichia coli and purified by affinity chromatography. Mice were immunized with the recombinant antigen, and hybridomas secreting MAbs were screened. Three stable hybridomas clones were selected and named 5D6, 7G3, and 8C9. The MAbs-based ELISA was scrutinized for species-specific recognition with a number of human throat swab samples from Group I (156 patients with typical respiratory illness clinically confirmed before) and Group II (57 healthy donors).

RESULTSIn Group I, 55 positive cases were detected by anti-EB MAb-based ELISA, 51 cases were positive by MAbs 5D6-based ELISA, and 33 and 38 cases were positive by MAb 8C9 and 7G3-based ELISA respectively. Of the 57 samples from Group II "healthy donors", 5 were positive and 52 were negative with both anti-EB and 5D6-based tests, while 2 and 3 positive cases were identified by the other two MAb-based ELISAs respectively.

CONCLUSIONThe novel MOMPVD2-VD3 MAb-based assay may have higher specificity than the anti-EB MAb, which may possibly be used as an alternative tool for the diagnosis of C. pneumoniae infection.

Animals ; Antibodies, Monoclonal ; Bacterial Outer Membrane Proteins ; immunology ; Chlamydophila Infections ; diagnosis ; microbiology ; Chlamydophila pneumoniae ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Mice ; Protein Structure, Tertiary

Adult ; Antibodies, Monoclonal ; metabolism ; Diagnosis, Differential ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; pathology ; surgery ; Male ; MyoD Protein ; metabolism ; Pneumonectomy ; Pulmonary Veins ; metabolism ; pathology ; surgery ; S100 Proteins ; metabolism ; Sarcoma, Alveolar Soft Part ; metabolism ; pathology ; surgery ; Vascular Neoplasms ; metabolism ; pathology ; surgery

In this paper, based on the special influence of supramolecular chemistry on the basic theory of Chinese medicines ( CM) , the authors further analyzed the history of meridian tropism and natural origins of CM organisms and explained CM ingredients and the universal regularity of the automatic action of the supramolecular "imprinting templates" hole channel structure. After entering human bodies, CMs, as the aggregation of supramolecular "imprinting templates" , automatically seek supramolecular subjects that are matched with their "imprinting templates" in human meridians and organs for the purpose of self-recognition, self-organization, self-assembly and self-replication, so as to generate specific efficacy in meridians and organs, which is reflected as the meridian tropism phenomena at macro level. This regularity can be studied by in vitro and in vivo experimental studies. In vitro methods are mostly supra molecular structure analysis and kinetic and thermodynamic parameter calculation; Whereas in vivo methods are dominated by the analysis on object component distribution, chromatopharmacodynamic parameters and network chromatopharmacodynamic parameters; Particularly, the acupoint-medicine method can simplify to study the supramolecular subject-object relations. Consequently, CM's'meridian tropism reveals the universal regularity for interactions of macromolecular and micromolecular "imprinting templates" of subjects and objects in natural organisms. As the first barrier for the material base of the CM theory and breakthrough in the modernization of the basic CM theory, meridian tropism plays an important role in studies on basic theories of the basic CM theory.
Acupuncture Points ; Drug Therapy ; Drugs, Chinese Herbal ; chemistry ; Humans ; Medicine, Chinese Traditional ; Meridians

The study was purposed to analyze DNA and allele structure of the partial D phenotypes D(Va) and D(VI) of the Rhesus blood group in Chinese. Through polymerase chain reaction (PCR) and direct genomic DNA sequencing, the RHD gene was detected in three weak D individuals identified serologically. The results showed that among the three weak D individuals, one was identified as partial D phenotype D(Va) (Hus) type and genotyped DccEe; another two were testified as D(VI) III type and genotyped DCcee. Moreover, the breakpoints of the replaced region by RHCE in D(Va) (Hus) were 5' end of the exon 5 and 3' end of the intron 5, and there were 7 novel polymorphisms in intron 5: 23-25(GCA)2, 98G>A, 168-169insG, 205-206insT, 494-495insA, 1256-1257insC, 1347G>T. In conclusion the whole exon 5 and intron 5 are replaced by RHCE in D(Va) (Hus) detected in Chinese.
China ; Exons ; genetics ; Genotype ; Humans ; Introns ; genetics ; Phenotype ; Polymerase Chain Reaction ; Rh-Hr Blood-Group System ; genetics ; Sequence Analysis, DNA

OBJECTIVETo study the proliferation and differentiation of neural stem cells in the subventricular zone (SVZ) in neonatal rats after bilateral common arteries occlusion.

METHODSNinety-six 3-day-old Sparuge-Dawley rats were randomly divided into two groups: ischemia and control. Rats in the ischemia group were subjected to bilateral common arteries occlusion and the rats in the control group were sham-operated. All rats were administrated with 5-bromodeoxyuridine (BrdU) (50 mg/kg) via intraperitoneal injection. Rats were sacrificed and their brains were removed 1, 4, 7, 10, 14 and 35 days after ischemia. Using brain paraffin sections and immunofluorescence assays, the number of newborn cells in the SVZ was counted. Newborn neural stem cells and oligodendrocytes in the SVZ were observed, and then double marked with BrdU and nestin or osmium tetroxide (O4).

RESULTSThe number of BrdU+ cells (neural stem cells) in the SVZ in the ischemia group was greater than in the control group 4, 7, 10 and 14 days after ischemia, and reached a peak at 4 days after ischemia (253.1+/- 49.3 vs 133.5+/- 17.7; P< 0.01). By 35 days after ischemia, the number of BrdU+/O4+ cells (oligodendrocytes) in the corpus callosum (56.0+/- 7.2 vs 17.0+/- 6.4; P< 0.01), the septal nuclei (45.0+/- 11.9 vs 20.5+/- 5.0; P< 0.01), the striatum (34.5+/- 4.2 vs 14.5+/- 5.8; P< 0.01) and the olfactory bulb (46.5+/- 6.6 vs 23.5+/- 8.4; P< 0.01) in the ischemia group increased significantly as compared to the control group (P< 0.01).

CONCLUSIONSBrain ischemia can activate the proliferation of neural stem cells in the SVZ and promote neural stem cells differentiation into oligodendrocytes. The immature brain may have the capacity for self-repair after ischemic brain injury.

Animals ; Animals, Newborn ; Brain Ischemia ; physiopathology ; therapy ; Bromodeoxyuridine ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cerebral Ventricles ; physiopathology ; Female ; Male ; Neurogenesis ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation