Objective To investigate the interaction between a novel antimicrobial compound,HL-J6,and penicillin-binding protein 1(PBP1)of Staphylococcus aureus.Methods With MRSA252 genomic DNA as the template and PBP1F and PBP1R as primers,the expression plasmid pET30a-pbp1-39-608 was constructed by amplifying the target gene fragment followed by cloning into the Nde I/Xho I restriction sites of the pET30a vector.Then the obtained plasmids were transformed into Escherichia coli for the expression of PBP1-39-608 protein,and the product was purified by affinity chromatography.The inhibitory effect of HL-J6 on the transpeptidase activity of PBP1-39-608 was measured using peptidoglycan side chain backbone peptide,with thiol ester analog S2d as the substrate.The affinity between HL-J6 and PBP1-39-608 was detected using microscale thermophoresis(MST),and the binding interaction was confirmed by cellular thermal shift assay(CETSA).Molecular docking and dynamics simulation were performed using AutoDock Vina and Desmond software,respectively,to elucidate the binding mode of HL-J6 with the PBP1-39-608 protein and the key amino acid residues involved.Results The recombinant plasmid pET30a-pbp1-39-608 was successfully constructed,and PBP1-39-608 protein was produced after induction and purified,yielding a protein with an approximate molecular mass of 65×103.HL-J6 inhibited the transpeptidase activity of PBP1-39-608 in a time-dependent manner(P<0.001).The dissociation constant Kd of the binding between HL-J6 and PBP1-39-608 was 64.92 μmol/L.Molecular docking results showed that HL-J6 bound to the active pocket of PBP1-39-608 by interacting with key residues such as ILE-348,ASN-370,THR-516 and PHE-423,with a binding score of-8.38 kcal/mol(<-5.00 kcal/mol).Dynamics simulation results indicated that the complex became stable after 50 ns.Conclusion HL-J6 effectively inhibits the transpeptidase activity of Staphylococcus aureus PBP1,and shows stable interaction with the protein.

OBJECTIVE To explore the effect and mechanism of the alcoholic extract from Scabiosa comosa against hepatic fibrosis （HF）. METHODS Intragastrical administration of carbon tetrachloride was given to induce HF model. By observing the pathological changes in liver tissue， mRNA and protein expressions of HF indexes [α-smooth muscle actin （α-SMA）， collagen type Ⅰ] and phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） pathway-related factors were detected， and the improvement effects and possible mechanism of low-dose， medium-dose and high-dose （50， 100， 200 mg/kg） of alcoholic extract from S. comosa on HF model rats were investigated. Drug-containing serum was prepared by intragastrical administration of alcoholic extract from S. comosa at a concentration of 1 800 mg/（kg·d） （calculated by the amount of raw material）. The effects of drug- containing serum of alcoholic extract from S. comosa on the expression of miRNA-21 were observed through the intervention of HSC-T6 cells with low， medium and high concentrations of drug-containing serum of alcoholic extract from S. comosa （diluted to 10%， 15%， 20%）. miRNA-21 mimics or inhibitors were used to transfect HSC-T6 cells， and the mRNA and protein expressions of factors related to the PI3K/Akt signaling pathway were detected. RESULTS The results of in vivo experiments showed that low， medium and high doses of alcoholic extract from S. comosa significantly ameliorated the histopathological changes in liver tissue of HF rats， and the percentage of collagen was significantly reduced （P＜0.01）； mRNA and protein expressions of the indicators related to HF as well as PI3K and Akt were significantly reduced （P＜0.01）， and mRNA and protein expressions of phosphatase and tensin homolog deleted on chromosome ten （PTEN） were increased in liver tissue of rats （P＜0.01）. The results of in vitro experiments showed that drug-containing serum of alcoholic extract from S. comosa significantly inhibited the expression of miRNA-21 at low， medium and high concentrations （P＜0.01）； whereas after transfection with miRNA-21 mimics， it was found that miRNA-21 mimics significantly increased mRNA and protein expressions of PI3K and Akt （P＜0.01）， while significantly decreased mRNA and protein expressions of PTEN （P＜0.01）； after transfection with miRNA-21 inhibitor， the changes of above indexes were opposite to the above results （P＜0.01）. CONCLUSIONS Alcoholic extracts of S. comosa may inhibit the PI3K/Akt signaling pathway by affecting the expression of miRNA-21， so as to achieve the effect of anti-hepatic fibrosis.

Objective To construct and evaluate the rat model of thyroid nodule liver-qi depression syndrome with the combination of disease and syndrome,so as to lay a foundation for the exploration of the mechanism and clinical research of traditional Chinese medicine in the treatment of thyroid nodule.Methods 36 rats were randomly divided into three groups.The model was induced by chronic unpredictable mild stress(CUMS)stimulation combined with propylthiouracil(PTU).The rats in the blank group were fed with saline at the dose of 1 mL·100 g-1·d-1,and the rats in the PTU group were fed with 0.1%PTU solution at the dose of 1 mL·100 g-1·d-1.The rats in the CUMS+PTU group were fed with CUMS stimulation in a single cage and intragastric administration of 0.1%PTU solution according to the dose of 1 mL·100 g-1·d-1,and the modeling time lasted for 6 weeks.After the establishment of the model,the model of combination of disease and syndrome was evaluated comprehensively by means of animal behavior detection,ultrasonic image,apparent morphology observation,pathological staining and serum hormone index detection.Results Compared with control group and PTU group,the total distance in the open field was significantly shortened,the sucrose preference rate was significantly decreased,and the swimming suspension time was significantly prolonged in the CUMS+PTU group,showing obvious symptoms of liver qi stagnation.In addition,compared with the other two groups,the thyroid gland of CUMS+PTU group showed obvious enlargement and nodule formation,thyroid index significantly increased,and thyroid follicular epithelial cells showed a high phase of proliferation.In terms of hormone levels,compared with control group,the serum levels of free triiodothyronine(FT3)and free thyroxine(FT4)in CUMS+PTU group were significantly lower,while the level of thyroid stimulating hormone(TSH)was significantly higher,which was in line with the current evaluation criteria of thyroid nodule model.Conclusion The method of CUMS stimulation combined with PTU drug induction can successfully prepare the rat model of thyroid nodule liver-qi depression syndrome,and the comprehensive evaluation method is stable and reliable,which is helpful to improve the scientific nature of the disease-syndrome combination model.

Objective:To analyze the clinical data of 203 discharged patients with corona virus disease 2019(COVID-19), and to investigate the predictors for the severe cases.Methods:Confirmed COVID-19 cases hospitalized at Zhongnan Hospital of Wuhan University from January 1 to February 1, 2020 were consecutively enrolled, who were divided into severe group and non-severe group.The clinical data of enrolled patients were collected and the clinical manifestations, laboratory results, imaging, treatments and prognosis of patients in the two groups were analyzed. Mann-Whitney U rank sum test and chi-square test were used for statistical analysis. Results:A total of 203 discharged patients with COVID-19 were enrolled. The common clinical manifestations included fever (89.2%, 181/203), dry cough (60.1%, 122/203), chest distress (35.5%, 72/203), shortness of breath(29.1%, 59/203)and myalgia or arthralgia (26.6%, 54/203). The time from disease onset to hospital admission was 5.8 days (1.0 to 20.0 days). Among 203 enrolled patients, 107(52.7%) were divided into severe group and 96(47.3%) were non-severe group. The age in severe group was 60 years (23 to 91 years), which was significantly older than non-severe group (47 years (20 to 86 years)), the difference was statistically significant ( Z=-6.12, P<0.01). There were 63.6%(68/107) patients in severe group with at least one underlying disease, which was significantly more than non-severe group (20.8% (20/96)), the difference was statistically significant ( χ2=37.60, P<0.01). The proportions of patients with increased white blood cells, decreased lymphocytes and albumin, elevated alanine aminotransferase, aspartate aminotransferase, creatinine, lactic acid dehydrogenase, creatine kinase, fasting blood glucose, D-dimer, erythrocyte sedimentation rate, C-reactive protein, interleukin-6, and procalcitonin in severe group were all higher. On admission, 172 patients (84.7%) had bilateral patchy shadows or ground glass opacity in the lungs on chest imaging study, 20(9.9%) presented pleural effusion. Fifty-five cases (27.1%) showed progressions of lung lesions on computed tomography (CT) rescan at an average interval of five days. Among 203 patients, 123(60.6%) were given oxygen therapy upon admission, 107(52.7%) were given short-term glucocorticoid therapy, and 131(64.5%) received antiviral therapy; and 26(12.8%) died. The hospital stay was 11.0 days (1.0 to 45.0 days). Conclusions:Fever is the most common symptoms in COVID-19 patients.Elderly and patients with underlying diseases are risk factors for progression to severe cases. The elderly patients should be strengthened early monitoring, paid attention to the control of underlying diseases, and reduce the occurrence of critical diseases.

This study was aimed to explore brain regions which were closely related to the disease onset of premenstrual syndrome (PMS) with liver-qi depression. The BOLD-functional magnetic resonance imaging (fMRI) was used in the study. The processing of imaging data was based on the SPM 8 software and the REST software of the matlab platform. Each cluster was more than 389 continuous voxel. The brain region with single voxel of P < 0.05 (corrected) was defined as region with statistical significance. The 2 Sample T-Test was applied in the case group and the control group. The results showed that compared with the normal control group, the frontal lobe, occipital lobe, insula, limbic lobe, basal nuclei, and cingulate gyrus were activated in the PMS with liver-qi depression cases. It was concluded that the disease onset of PMS with liver-qi depression cases was related to brain regions such as frontal lobe, occipital lobe, insula, limbic lobe, basal nuclei, and cingulate gyrus.

This study was aimed to reveal the roles of biological factors T3, T4 and TSH in the pathogenesis of liver-qi invasion syndrome and liver-qi depression syndrome of premenstrual syndrome (PMS) in the body. Thus, we may expound the connotation of conceptpotential stagnation ofqi and blood. Female workers of Jinan and Qingdao were selected as target groups by epidemiological cross-sectional survey. And specially trained investigators were responsible for screening patients referred to criterion on the international diagnostic standards and syndrome diagnosis of PMS. Patients' blood was taken as samples during the follicular phase (6 to 14 days), luteal phase (15 to 23 days), premenstrual phase (24 to 28 days) and the menstrual phase (1 to 5 days), respectively. Blood samples were disposed according to requirements of radioimmunoassay. Single factor analysis of variance was used in the comparison with the level of T3, T4 and TSH in menstrual blood for different time intervals of the case group and the normal group by SPSS 10.0 statistical software. The results showed that T3 levels of patients with either syndrome were significantly reduced during the premenstrual phase and the menstrual phase (P < 0.05). There were no significant changes on the level of serum T4 and TSH before and after the occurrence of the disease. Both of them were significantly increased during each phase of the menstrual cycle. It was concluded that changes of T3, T4 and TSH were one of the mechanisms in disease with liver-qi invasion syndrome and liver-qi depression syndrome. The micro change was the essence ofpotential stagnation of qi and blood, which was the body condition of this disease.

Start to explore the theory conditions around the theme why to build the modern basic theory of tradi-tional chinese medicine and what kind of theory we want to establish . On the basis of making clear the related concepts such as theory, knowledge, scientific theory and scientific knowledge, discuss the theoretical development pathways and evaluation criteria, compare the development mode of Chinese and Western medicine, providing a refer-ence for evaluation and future development of the modern basic theory of traditional chinese medicine. According to Carl G. Hempel's bridge principle, summarize the five bridge principles and a logic protection principle of the research and foundation of the modern basic theory of traditional chinese medicine, and explain its role and signifi-cance. Put forward a theoretical framework of the modern basic theory of traditional chinese medicine Three levels and five grades like a tree structure, demonstrate the scientific connotation of 11 principles and concepts under the frame. Analyze and demonstrate its scientific rationality according to the role and evaluation criteria of scientific theory mentioned above, clearing direction for future development.

Around the core issue how to evaluate and test hypothesis, starting from the concept of hypothesis and scientific hypothesis, learning form recent progress and understanding at home and abroad, we have analyzed the type of scientific hypothesis, test methods and pre-mortem and post-mortem evaluation criteria in depth, established scientific vision and discipline ruler, explored the awareness level of the Chinese medicine hypothesis, pointed out the problems and gaps. Whereby, we have analyzed and demonstrated brain regulating five internal organs concept of wholism scientific hypothesis, and proven it belongs to the scientific hypothesis. We were confident that these works mentioned above would lead the theoretical reform changing Chinese medicine hypothesis from traditional description to the nature clarification.

ObjectiveTo explore the epidemiologic features and distribution pattems of hepatitis C virus (HCV) genotype infection among HIV positive former blood donors (FBDs) and transfusion recipients in Hubei province.Methods597 serum samples from HIV-positive patients in Hubei were collected and examined for anti-HCV by enzyme-linked immunosorbent assay ( ELISA ).Reverse transcription nested polymerase chain reaction (RT-nested PCR) amplification and DNA sequencing were used to evaluate the HCV core regions.ResultsThe prevalence rates of HCV in HIV positive FBDs and transfusion recipients were 76.5％ (205/268) and 57.4％ (189/329) respectively.HCV genotypes 1b (92.8％,90/97) and 2a (7.2％,7/97 ) were detected.ConclusionsBlood donation and blood transfusion are the major modes of HIV-HCV co-infection in Hubei province.The prevalence of HCV in HIV positive transfusion recipients is lower than that in HIV positive FBDs.HCV genotypes 1b and 2a are the predominant strains among HIV-positive FBDs and transfusion recipients.

ObjectiveTo compare the mRNA and protein expression levels ofγ-aminobutyric acid receptor B2 ( GABABR2 ) in brain regions of male rats with high level aggressive behaviors and low level aggressive behaviors respectively,and provide clues for exploring mechanism of GABA in aggressive behaviors.MethodsWistar male rats were randomly divided into two groups:the normal group and the aggressive behavior group.Then social isolation and resident intruder stresses were used to establish high level and low level aggressive behavior in the aggressive behavior group.The mRNA and protein level of GABABR2 in parietal cortex,prefrontal cortex,hypothalamus and hippocampus of the three groups rats ( n=10 in each group) were detected using RT-PCR and Western blot respectively.ResultsThe GABABRB22 RT-PCR/Western blot relative integrated optical density of parietal cortex,prefrontal cortex,hypothalamus and hippocampus in the normal group rats respectively were.Those of the above four brain regions in high aggressive behavior group rats respectively were ( 0.507 ± 0.049/0.626 ±0.038 ),(0.609 ± 0.049/0.652 ± 0.010 ),( 0.359 ± 0.030/0.731 ± 0.044 ) and ( 0.296 ± 0.054/0.452 ±0.079) were significantly lower (P＜0.05) compared with the normal group rats.In the low aggressive behavior group rats,the GABABRB2 RT-PCR/Western blot relative integrated optical density of parietal cortex and hippocampus increased statistically(P＜ 0.05 ),while those of prefrontal cortex and hypothalamus decreased obviously (P ＜ 0.05 ).all in comparison with the normal group rats.Conclusion Different expression levels of GABABR2 in parietal cortex,prefrontal cortex,hypothalamus and hippocampus are relative to aggressive behaviors,which might be one of the mechanism for GABA in aggressive behaviors.