Jujubae Fructus alleviates intestinal injury caused by toxic medicinals in Shizao Decoction based on correlation between intestinal flora and host metabolism / 中国中药杂志

Genkwa Fols, Kansui Radix, and Euphorbiae Pekinensis Radix in Shizao Decoction(SZD) are toxic to intestinal tract. Jujubae Fructus in this prescription can alleviate the toxicity, but the mechanism is still unclear. Therefore, this study aims to explore the mechanism. To be specific, 40 normal Sprague-Dawley(SD) rats were classified into the normal group, high-dose and low-dose SZD groups, and high-dose and low-dose SZD without Jujubae Fructus(SZD-JF) groups. The SZD groups were given(ig) SZD, while SZD-JF groups received the decoction without Jujubae Fructus. The variation of body weight and spleen index were recorded. The patho-logical changes of intestinal tissue were observed based on hematoxylin and eosin(HE) staining. The content of malondialdehyde(MDA) and glutathione(GSH) and activity of superoxide dismutase(SOD) in intestinal tissue were measured to evaluate the intestinal injury. Fresh feces of rats were collected to detect intestinal flora structure by 16S ribosomal RNA gene(16S rDNA) sequencing technology. The content of fecal short chain fatty acids and fecal metabolites was determined by gas chromatography-mass spectrometer(GC-MS) and liquid chromatography-mass spectrometer ultra-fast liquid chromatography-quadrupole-time-of-flight mass spectrometer(UFLC-Q-TOF-MS), separately. Spearman's correlation analysis was employed to analyze the differential bacteria genera and differential metabolites. RESULTS:: showed that high-dose and low-dose SZD-JF groups had high content of MDA in intestinal tissue, low GSH content and SOD activity, short intestinal villi(P<0.05), low diversity and abundance of intestinal flora, variation in the intestinal flora structure, and low content of short chain fatty acids(P<0.05) compared with the normal group. Compared with high-dose and low-dose SZD-JF groups, high-dose and low-dose SZD groups displayed low content of MDA in intestinal tissue, high GSH content and SOD activity, recovery of the length of intestinal villi, increased abundance and diversity of intestinal flora, alleviation of dysbacteria, and recovery of the content of short chain fatty acids(P<0.05). According to the variation of intestinal flora and fecal metabolites after the addition of Jujubae Fructus, 6 differential bacterial genera(Lactobacillus, Butyricimonas, Clostridia＿UCG-014, Prevotella, Escherichia-Shigella, Alistipes),4 differential short chain fatty acids(such as acetic acid, propionic acid, butyric acid, valeric acid) and 18 differential metabolites(such as urolithin A, lithocholic acid, and creatinine) were screened out. Beneficial bacteria such as Lactobacillus were in positive correlation with butyric acid and urolithin A(P<0.05). The pathogenic bacteria such as Escherichia-Shigella were in negative correlation with propionic acid and urolithin A(P<0.05). In summary, SZD-JF caused obvious intestinal injury to normal rats, which could lead to intestinal flora disorder. The addition of Jujubae Fructus can alleviate the disorder and relieve the injury by regulating intestinal flora and the metabolites. This study discusses the effect of Jujubae Fructus in relieving the intestinal injury caused by SZD and the mechanism from the perspective of intestinal flora-host metabolism, which is expected to serve as a reference for clinical application of this prescription.

Overexpression of chicken Klf2 promotes klf7 transcription and inhibits adipocyte differentiation / 生物工程学报

Overexpression of Krüppel like factor 2 (Klf2) or Klf7 inhibits adipocyte formation. However, it remains unclear whether Klf2 regulates klf7 expression in adipose tissue. In this study, oil red O staining and Western blotting were employed to study the effect of Klf2 overexpression on the differentiation of chicken preadipocytes. The results showed that Klf2 overexpression inhibited the differentiation of chicken preadipocytes induced by oleate and the expression of pparγ, while promoted klf7 expression in chicken preadipocytes. Spearman correlation analysis was used to study the correlation between the expression data of klf2 and klf7 in the adipose tissue of both human and chicken. The results showed that there was a significantly positive correlation between the expression of klf2 and klf7 in adipose tissues (r > 0.1). Luciferase reporter assay showed that overexpression of Klf2 significantly promoted the activity of chicken klf7 promoter (-241/-91, -521/-91, -1 845/-91, -2 286/-91, -1 215/-91; P < 0.05). In addition, the activity of klf7 promoter (-241/-91) reporter in chicken preadipocytes was significantly positively correlated with the amount of klf2 overexpression plasmid transfected (Tau=0.917 66, P=1.074×10-7). Moreover, Klf2 overexpression significantly promoted the mRNA expression of klf7 in chicken preadipocytes (P < 0.05). In conclusion, upregulation of klf7 expression might be one of the pathways that Klf2 inhibits chicken adipocyte differentiation, and the sequence from -241 bp to -91 bp upstream chicken klf7 translation start site might mediate the regulation of Klf2 on klf7 transcription.

Mitochondrial aldehyde dehydrogenase 2 alleviates septic liver injury by inhibiting ferroptosis in mouse model / 中华危重病急救医学

OBJECTIVE@#To observe the ferroptosis triggered by in different pathways during cecal ligation and puncture (CLP)-induced liver injury in septic mice, and to investigate whether mitochondrial aldehyde dehydrogenase 2 (ALDH2) can alleviate sepsis-induced liver injury by inhibiting ferroptosis.@*METHODS@#Sixty 8-week-old male C57BL/6J mice were randomly divided into sham operation group (Sham group), CLP group, ferroptosis inhibitor ferrostain-1 (Fer-1) group, ALDH2-specific agonist Alda-1 group, iron chelator deferasirox Fe3+ chelate (DXZ) group and dimethyl sulfoxide (DMSO) group, with 10 mice in each group. The septic liver injury was induced by CLP in mice model. In the Sham group, only laparotomy was performed without ligation and puncture of the cecum. 10 mL/kg 5% DMSO, 5 mg/kg Fer-1, 50 mg/kg DXZ and 10 mg/kg Alda-1 were injected intraperitoneally 1 hour before CLP in the DMSO, Fer-1, DXZ and Alda-1 groups respectively. At 24 hours after operation, eyeball blood and liver tissue were collected from anesthetized mice. The hepatic structure and inflammatory infiltration were observed by hematoxylin-eosin (HE) staining. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) in serum, the levels of hepatic malondialdehyde (MDA), superoxide dismutase (SOD) and reactive oxygen species (ROS) were detected. Western blotting was used to detect the protein expressions of ALDH2, ferroptosis-related proteins glutathione peroxidase 4 (GPX4), ferroptosis suppressor protein 1 (FSP1) and transferrin receptor 1 (TFR1) in liver tissue.@*RESULTS@#Compared with Sham group, the mice in CLP group showed varying degrees of congestion, disorganized hepatocyte arrangement, inflammatory cell infiltration at 24 hours after operation. Compared with the CLP group, the mice in the Fer-1 group, DXZ group and Alda-1 group liver morphology, liver injury and inflammatory cell infiltration was improved. Compared with Sham group, the serum levels of ALT and AST, the contents of MDA and ROS, and the expression of TFR1 protein in CLP group were significantly increased, while the activity of SOD and the expressions of ALDH2, GPX4 and FSP1 protein in CLP group were significantly decreased. Compared with CLP group, serum ALT and AST levels in Fer-1, DXZ and Alda-1 groups were significantly decreased [ALT (U/L): 45.76±10.81, 37.30±2.98, 36.40±12.75 vs. 73.06±12.20, AST (U/L): 61.57±2.69, 52.41±6.92, 56.05±8.29 vs. 81.59±5.46, all P < 0.05], and the contents of MDA, ROS and TFR1 protein expression in liver tissue were significantly decreased [MDA (μmol/L): 0.60±0.10, 0.57±0.18, 0.83±0.39 vs. 1.61±0.30, ROS (fluorescence intensity): 270.34±9.64, 276.02±62.33, 262.05±18.55 vs. 455.38±36.07, TFR1/GAPDH: 0.90±0.04, 1.01±0.09, 0.55±0.08 vs. 1.18±0.06, all P < 0.05], and the SOD activity and ALDH2, GPX4 and FSP1 protein expressions in liver tissue were significantly increased [SOD (kU/g): 88.77±8.20, 88.37±4.47, 93.43±7.24 vs. 50.27±3.57, ALDH2/GAPDH: 1.10±0.15, 1.02±0.07, 1.14±0.07 vs. 0.70±0.04, GPX4/GAPDH: 1.02±0.12, 0.99±0.08, 1.05±0.19 vs. 0.71±0.10, FSP1/GAPDH: 1.06±0.24, 1.02±0.08, 0.93±0.09 vs. 0.66±0.03, all P < 0.05]. There was no significant difference in the parameters between DMSO group and CLP group.@*CONCLUSIONS@#Both GPX4 and FSP1 mediated ferroptosis are involved in liver injury in septic mice. Activation of ALDH2 and inhibition of ferroptosis can alleviatehepatic injury. ALDH2 may play a protective role by regulating FSP1 and GPX4 mediated ferroptosis.

The development trend and hotspots analysis of international genetics research of the coronavirus from 2003 to 2021 / 中华医学科研管理杂志

Objective:To analyze the overall layout of coronavirus genetics research in the past 20 years from the perspective of publication time, journals, keywords, citations and funds, identify core research institutions and their cooperation networks in this field, and explore current research hotspots.Methods:PubMed and Web of Science databases were searched separately for the research literature and citations data about coronavirus genetics from 2003 to 2021, and Excel was used to analyze the distribution of the literature and institutions, and then the Citespace software was selected for institutional cooperation network and reference co-citation cluster analysis.Results:The literature about coronavirus genetics had increased significantly in 2020. The top 10 journals have collected 23.4% of relevant literature. Chinese Academy of Sciences has published the most documents in this field, in the top 10 high-yielding institutions, 7 are from China. In the cooperation network, the Chinese Academy of Sciences, University of Hong Kong and University of Sao Paulo have high betweenness centrality. The total litation times and average citation times of funded papers were higher than those of non-funded papers. There have been 7 research hotspots in the past 20 years. The research on " the gene sequence and functional receptors of the Middle East respiratory syndrome coronavirus" , " the gene traceability and drug development of the SARS-Cov-2" as well as " genotyping, origin and economic impact of paroxysmal porcine epidemic diarrhea virus in the United States" are still active in the past five years.Conclusions:Coronavirus genetics research will be at a sustained high level in the next few years or even longer. It is difficult to publish papers in journals with high impact factors (IF>5.00) in related fields. Chinese research institutions are active in this field. The Chinese Academy of Sciences and the University of Hong Kong are the most influential, and they have the closest cooperation with other institutions. The genetic tracing, gene sequence and functional receptor of coronavirus and drug development are likely to be the forefront of the research in this field.our govermment should advance the layout in this field and increase the number of research funds and financial support.

Inhibiting ferroptosis attenuates myocardial injury in septic mice: the role of lipocalin-2 / 南方医科大学学报

OBJECTIVE@#To explore the contribution of ferroptosis to myocardial injury in mouse models of sepsis and the role lipocalin-2 (Lcn2) in ferroptosis.@*METHODS@#Adult male C57BL/6 mice were randomized equally into sham-operated group, cecal ligation and puncture (CLP)-induced sepsis group, and CLP + Fer-1 group where the mice received intraperitoneal injection of 5 mg/mL Fer-1 (5 mg/kg) 1 h before CLP. The left ventricular functions (including LVEF%, LVFS%, LVIDd and LVIDs) of the mice were assessed by echocardiography at 24 h after CLP. Myocardial injury in the mice was observed with HE staining, and the changes of myocardial ultrastructure and mitochondria were observed using transmission electron microscopy (TEM). Serum TNF-α level was measured with ELISA, and the changes of myocardial iron content were detected using tissue iron kit. The protein expressions of myocardial Lcn2, glutathione peroxidase 4 (GPX4) and ferroptosis suppressor protein 1 (FSP1) were determined with Western blotting.@*RESULTS@#The septic mice showed significantly decreased LVEF%, LVFS% and LVIDd and increased LVIDs at 24 h after CLP (P < 0.05), and these changes were significantly improved by Fer-1 treatment. Sepsis caused obvious myocardial pathologies and changes in myocardial ultrastructure and mitochondria, which were significantly improved by Fer-1 treatment. Fer-1 treatment also significantly ameliorated sepsis-induced elevations of serum TNF-α level, myocardial tissue iron content, and Lcn2 protein expression and the reduction of GPX4 and FSP1 protein expression levels (P < 0.05).@*CONCLUSION@#GPX4- and FSP1-mediated ferroptosis are involved in myocardial injury in mice with CLP-induced sepsis, and inhibition of ferroptosis can attenuate septic myocardial injury, in which Lcn2 may play a role.

The clinical phenotype and gene analysis of syndromic deafness with PTPN11 gene mutation / 中华耳鼻咽喉头颈外科杂志

Objective: To analyze the clinical phenotype and screen the genetic mutations of hereditary deafness in three deaf families to clarify their molecular biology etiology. Methods: From January 2019 to January 2020, three deaf children and family members were collected for medical history, physical examination, audiology evaluation, electrocardiogram and cardiac color Doppler ultrasound, temporal bone CT examination, and peripheral blood DNA was obtained for high-throughput sequencing of deafness genes. Sanger sequencing was performed to verify the variant sites among family members. The pathogenicity of the variants was evaluated according to the American College of Medical Genetics and Genomics. Results: The probands in the three families had deafness phenotypes. In family 1, proband had multiple lentigines, special facial features, growth retardation, pectus carinatum, abnormal skin elasticity, cryptorchidism and other manifestations. In family 2, proband had special facial features, growth retardation and abnormal heart, and the proband in family 3 had growth retardation and abnormal electrocardiogram. Genetic testing of three families detected three heterozygous mutations in the PTPN11 gene: c.1391G>C (p.Gly464Ala), c.1510A>G (p.Met504Val), c.1502G>A (p.Arg501Lys). All three sites were missense mutations, and the mutation sites were highly conserved among multiple homologous species. Based on clinical manifestations and genetic test results, proband 1 was diagnosed with multiple lentigines Noonan syndrome, and probands 2 and 3 were diagnosed with Noonan syndrome. Conclusion: Missense mutations in the PTPN11 gene may be the cause of the disease in the three deaf families. This study enriches the clinical phenotype and mutation spectrum of the PTPN11 gene in the Chinese population.

Dosimetric Analysis of Template-assisted 192Ir-source Hypofractionated Stereotactic Ablative Brachytherapy for Peripheral Lung Cancer / 肿瘤防治研究

Objective To explore the dose of template-assisted 192Ir source hypofractionated stereotactic brachytherapy (SABT) for peripheral lung cancer. Methods We retrospectively analyzed the dose parameters of GTV and OARs of 28 peripheral lung cancer patients treated with template-assisted 192Ir-source hypofractionated SABT, and compared the dose parameters between SABT with virtual SBRT. Results The Dmean and V150 for the GTV in the SABT plan were significantly higher than those in the SBRT plan (all P < 0.01). For OARs, all dosimetric parameters in the SABT plan were significantly lower than those in the SBRT plan (all P < 0.01), except for the D1000cm3 and D1500cm3 for the lung (P > 0.05). Conclusion Template-assisted 192Ir source hypofractionated SABT ensures high dose in the gross tumor volume and reduces the dose in organs at risk in the treatment of peripheral lung cancer.

Accuracy of Nolla Method for Age Estimation of Northern Chinese Han Children / 法医学杂志

Objective To study the accuracy of Nolla method for age estimation of Northern Chinese Han children aged between 5.00 and 14.99 years based on original transformation tables and multiple regression model. Methods A total of 2 000 orthopantomographs （OPGs） were collected from the Hospital of Stomatology, Xi'an Jiaotong University, including 1 000 males and 1 000 females. Development stage of 7 left mandibular permanent teeth （except third molars） was assessed based on Nolla method, then age estimation was conducted through transformation tables and multiple regression model, respectively. Firstly, the development stage results of 7 permanent teeth were added up and the estimated age was obtained through the original transformation tables. Secondly, 80% of the samples （80 males and 80 females in each age group） were randomly selected from 2 000 OPGs as the train set. The chronological age of the selected patients was taken as the dependent variable, while gender and the development stage results of 7 permanent teeth were taken as the independent variable to establish multiple regression model. The remaining 20% of the samples were substituted into the model as the test set, to verify the accuracy of age estimation by multiple regression model. Results Mean chronological ages of males and females were 10.03±0.09 years and 10.01±0.09 years, respectively. The age estimated by original transformation tables showed an overestimation for males （0.18 years on average） and an underestimation for females （0.02 years on average）, with mean absolute error （MAE） of 0.94 years and 0.97 years, respectively. While the results by multiple regression model showed that males were overestimated by 0.06 years on average and females were underestimated by 0.02 years on average. The MAE was 0.66 years and 0.77 years, respectively. Conclusion The Nolla method is suitable for age estimation of Northern Chinese Han children. Compared with the original transformation tables method, the multiple regression model is more accurate for age estimation.

Clinical features and prognosis of children with acute leukemias of ambiguous lineage under different diagnostic criteria / 中国当代儿科杂志

OBJECTIVES@#To study the clinical features and prognosis of children with acute leukemias of ambiguous lineage (ALAL) under different diagnostic criteria.@*METHODS@#A retrospective analysis was performed on the medical data of 39 children with ALAL who were diagnosed and treated from December 2015 to December 2019. Among the 39 children, 34 received treatment. According to the diagnostic criteria for ALAL by World Health Organization and European Group for the Immunological Characterization of Leukemias, the 39 children were divided into two groups: ALAL group (@*RESULTS@#The 34 children receiving treatment had a 3-year event-free survival (EFS) rate of 75%±9% and an overall survival rate of 88%±6%. The children treated with acute myeloid leukemia (AML) protocol had a 3-year EFS rate of 33%±27%, those treated with acute lymphoblastic leukemia (ALL) protocol had a 3-year EFS rate of 78%±10%, and those who had no remission after induction with AML protocol and then received ALL protocol had a 3-year EFS rate of 100%±0% (@*CONCLUSIONS@#ALL protocol has a better clinical effect than AML protocol in children with ALAL, and positive MRD after induction therapy suggests poor prognosis. Hyperleukocytosis and adverse genetic changes are not observed in children with myeloid expression, and such children tend to have a good prognosis, suggesting that we should be cautious to take it as ALAL in diagnosis and treatment.

Research progress on pyroptosis and cardiovascular diseases / 生理学报

Pyroptosis is a form of programmed cell death which is closely related to the inflammatory response, mediated by Gasdermin protein and depends on the activity of cysteine aspartate specific protease (caspase). Pyroptosis is typically characterized by swelling and rupture of cell membrane, release of proinflammatory factors and cell contents from the plasma membrane to the extracellular environment, which aggravates inflammatory response. During the inflammatory response, NLRP3, caspase, Gasdermin D (GSDMD) and IL-1β play important roles in the occurrence and development of cardiovascular diseases. In this review, we focus on the role of pyroptosis in cardiovascular diseases including atherosclerosis, coronary heart disease, myocardial infarction, diabetic cardiomyopathy, pressure overload-induced ventricular remodeling and cardiac hypertrophy, myocarditis, arrhythmia and so on, and summarize the potential treatment targeting pyroptosis. It will provide the basis for prevention and treatment of clinical cardiovascular diseases.

Role of non-canonical pyroptosis in sepsis and other inflammatory diseases / 中南大学学报(医学版)

As a form of new programmed cell death, pyroptosis is divided into a canonical pyroptosis pathway and a non-canonical pyroptosis pathway. In recent years, it is reported that non-canonical pyroptosis is closely related to inflammatory reactions, which directly affects the occurrence, development, and outcome of sepsis, inflammatory bowel disease, respiratory disease, nerve system inflammatory disease, and other inflammatory diseases. When the cells were infected with Gram-negative bacteria or lipopolysaccharide (LPS), it can induce the activation of cysteinyl aspartate specific proteinase(caspase)-4/5/11 and directly bind to the cells to cleave gasdermin D (GSDM-D) into the active amino-terminus of GSDM-D. The amino-terminus of GSDM-D with membrane punching activity migrates to the cell membrane, triggering the rupture of the cell membrane, and the cell contents discharge, leading to the occurrence of non-canonical pyroptosis. After activation of caspase-11, it also promotes the canonical pyroptosis, activates and releases interleukin-1β and interleukin-18, which aggravated inflammation. Caspase-4/5/11, GSDM-D, Toll-like receptor 4 and high mobility group protein B1 are the key molecules of the non-canonical pyroptosis. Exploring the mechanisms of non-canonical pyroptosis and the related research progresses in inflammatory diseases intensively is of great significance for clinical prevention and treatment of the relevant diseases.

Effect of swertiamarin, gentiopicrin and sweroside on cell apoptosis and expression of Bcl-2 in rheumatoid arthritis fibroblast-like synoviocytes / 中国中药杂志

The aim of this paper was to discuss the effect of swertiamarin, gentiopicrin and sweroside on rheumatoid arthritis fibroblast-like synoviocytes(RA-FLSs) and B-cell lymphoma-2(Bcl-2) and their mechanisms. ZINC database and RCSB PDB database were retrieved for 3 D chemical structures of swertiamarin, gentiopicrin and sweroside and 3 D target protein structures. AutoDock Mgltools 1.5.6, AutoDockVina 1.1.2 and pyMOL 2.2.0 were applied for molecular docking to analyze the relationship between Bcl-2(1 GJH) target protein and important ingredients. The cell apoptosis of RA-FLSs was tested by Annexin V-FITC. The Bcl-2 protein expression of RA-FLSs treated with different ingredients was tested by Western blot. The Bcl-2 mRNA expression of RA-FLSs treated with different ingredients was tested by RT-PCR. Swertiamarin, gentiopicrin and sweroside were docked well with Bcl-2(1 GJH). The binding energy of swertiamarin was-6.9 kcal·mol~(-1), the binding energy of gentiopicrin was-6.7 kcal·mol~(-1) and the binding energy of sweroside was-6.4 kcal·mol~(-1). Compared with the blank group, the Bcl-2 protein expression of each group were reduced, while that of the gentiopicrin group was the highest(P<0.01). Compared with the blank group, the Bcl-2 mRNA expression of each groups were reduced. Gentiopicrin can reduce the Bcl-2 protein expression and the Bcl-2 mRNA expression, so as to promote the RA-FLSs apoptosis.

Correlation of plasma N-acetyl-neuraminic acid level with TIMI risk stratification and clinical outcomes in patients with acute coronary syndrome / 南方医科大学学报

OBJECTIVE@#To explore the correlation of plasma N-acetyl-neuraminic acid level with Thrombolysis In Myocardial Infarction (TIMI) risk score and clinical outcomes of patients with acute coronary syndrome (ACS).@*METHODS@#We consecutively enrolled 708 consecutive patients (401 male and 307 female, mean age 63.6±10.6 years) undergoing coronary angiography in our hospital between October, 2018 and July, 2019, including 597 patients with ACS and 111 without ACS (control group). The patients with ACS group were divided into high (=104), moderate (=425) and low (=68) risk groups according to their TIMI risk scores. All the participants were examined for plasma Neu5Ac level using liquid chromatography-tandem mass spectrometry and underwent coronary angiography with their Gensini scores calculated. The patients with ACS were followed up after discharge for a mean of 15 months for the occurrence of major adverse cardiac events (Mace). Binary logistic regression analysis was performed to identify the risk factors of Mace in these patients.@*RESULTS@#Plasma Neu5Ac levels were significantly higher in ACS group than in the control group ( < 0.05). ROC curve analysis showed that plasma Neu5Ac level could assist in the diagnosis of ACS (0.648 [0.597-0.699]) with a sensitivity of 39.2% and a specificity of 86.5% at the cutoff value of 288.50 ng/mL. In the ACS patients, plasma Neu5Ac level was significantly higher in the high-risk group than in the moderate-risk and low-risk groups ( < 0.05) and could assist in the diagnosis of a high risk (0.645 [0.588-0.703]) with a sensitivity of 42.3% and a specificity of 80.1% at the cutoff value of 327.50 ng/ mL. Plasma Neu5Ac was positively correlated with age, serum uric acid, creatinine, lipoprotein a, Ddimer, C-reactive protein, MB isoform of creatine kinase and Gensini score and negatively correlated with high-density lipoprotein level. During the followup, 80 ACS patients experienced Mace, who had significantly higher plasma Neu5Ac level than those without Mace (=517). Logistic regression analysis showed that plasma Neu5Ac level and a history of previous stroke were independent risk factors for the occurrence of Mace.@*CONCLUSIONS@#Plasma Neu5Ac level can provide assistance in the diagnosis and risk stratification of ACS and is an independent risk factor for prognosis of ACS patients.

In Silico Screening of Potential Chinese Herbal Medicine Against COVID-19 by Targeting SARS-CoV-2 3CLpro and Angiotensin Converting Enzyme II Using Molecular Docking / 中国结合医学杂志

OBJECTIVE@#To seek potential Chinese herbal medicine (CHM) for the treatment of coronavirus disease 2019 (COVID-19) through the molecular docking of the medicine with SARS-CoV-2 3CL hydrolytic enzyme and the angiotensin converting enzyme II(ACE2) as receptors, using computer virtual screening technique, so as to provide a basis for combination forecasting.@*METHODS@#The molecular docking of CHM with the SARS-Cov-2 3CL hydrolase and the ACE2 converting enzyme, which were taken as the targets, was achieved by the Autodock Vina software. The CHM monomers acting on 3CLpro and ACE2 receptors were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform, the active ingredients were selected, and the key CHMs and compounds were speculated. Based on the perspective of network pharmacology, the chemical-target network was constructed, and the functional enrichment analysis of gene ontology and the pathway enrichment analysis of Kyoto encyclopedia of genes and genomes were carried out by DAVID to speculate about the mechanism of action of the core drug pairs.@*RESULTS@#There are 6 small molecule compounds that have the optimal binding energy with the two target proteins. Among 238 potential anti-COVID-19 herbs screened in total, 16 kinds of CHM containing the most active ingredients, and 5 candidate anti-COVID-19 herbs that had been used in high frequency, as well as a core drug pair, namely, Forsythiae Fructus-Lonicerae Japonicae Flos were selected.@*CONCLUSION@#The core drug pair of Forsythiae Fructus-Lonicerae Japonicae Flos containing multiple components and targets is easy to combine with 3CLpro and ACE2, and exerts an anti-COVID-19 pneumonia effect through multi-component and multi-target, and plays the role of anti-COVID-19 pneumonia in multi-pathway.

Neuroprotective Effects of Low-molecular-weight Chondroitin Sulfate on Dopaminergic Neurons in MPTP-induced Parkinson ’s Disease Model Mice / 中国药房

OBJECTIVE： To observe neuroprotective effects of low-molecular-weight chondroitin sulfate （CS） on dopaminergic neurons in Parkinson’s disease （PD） mice model induced by 1-methyl-4-phenyl-1，2，3，6-tetrahydropyridine （MPTP）. METHODS： C57BL/6 mice were randomly divided into control group， MPTP injury group， low-molecular-weight CS low-dose and high-dose groups （100， 400 mg/kg）. Control group and MPTP injury group were given constant volume of normal saline intragstrically， administration groups were given relevant medicine intragastrically， once a day， for consecutive 17 d. Since 11th day after medication， except for control group， other groups were given MPTP solution （20 mg/kg） intraperitoneally to induce PD model， once a day， consecutive 5 d. After last medication， behavioral changes of mice （10 mice in each group） were evaluated by rotary rod fatigue tester. The damage of dopamine neurons （the percentage of TH positive cell and the percentage of fluorescence intensity） in substantia nigra of mice （3 mice in each group） was detected by immunohistochemistry and immunofluorescence. The content of dopamine in striatum was determined by HPLC （6 mice in each group）. The changes of oxidant stress indexes （SOD， GSH-Px， MDA） in substantia nigra of mice were determined by chemical colorimetry （6 mice in each group）. RESULTS： Compared with control group， retention time of mice on rotating rods was shortened significantly in MPTP injury group； TH positive cells of substantia nigra were decreased significantly， fluorescence intensity was obviously weakened； the percentage of positive cells and fluorescence intensity， the content of dopamine in striatum， the activities of SOD and GSH-Px in substantia nigra were decreased significantly， while the content of MDA was increased significantly （P＜0.01）. Compared with MPTP injury group， retention time of mice on the rotating rods was prolonged significantly in low-molecular-weight CS groups， the number of TH positive cells was increased significantly in substantia nigra and fluorescence intensity was increased significantly； the percentage of positive cells， the percentage of fluorescence intensity and the content of dopamine in striatum were increased significantly， while above indexes of high-dose group were significantly longer or higher than those of low-dose group （P＜0.05 or P＜0.01）. The activities of SOD and GSH-Px in substantia nigra were increased significantly in low-molecular-weight CS groups， while the content of MDA in substantia nigra was decreased significantly in low-molecular-weight CS high-dose group （P＜0.05 or P＜0.01）. CONCLUSIONS： Prophylactic administration of low-molecular-weight CS can relieve the damage of dopaminergic neurons in substantia nigra of PD model mice induced by MPTP in a dose-dependent manner， and increase the secretion of dopamine in striatum. The effect may be related to the inhibition of lipid peroxidation and the enhancement of antioxidant capacity of tissues.

Effect of Different Combinations of Gentianae Macrophyllae Radix on MMP-3 and TIMP-1 in Rheumatoid Arthritis Rats with Wind-cold-dampness Arthralgia / 中国实验方剂学杂志

Objective: To observe the effects of different combinations of Gentianae Macrophyllae Radix (Qinjiao) on the ankle joint matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) of rheumatoid arthritis (RA) model rats with wind-cold-dampness arthralgia.Method: Eighty healthy SD rats were randomly divided into 8 groups, namely blank control group, collage Ⅱ model group, wind-cold-dampness syndrome model group, positive control group, single-taste Gentianae Macrophyllae Radix group, Gentianae Macrophyllae Radix-Clematidis Radix et Rhizoma group (GC group), Gentianae Macrophyllae Radix-Taxilli Herba group (GT group), Gentianae Macrophyllae Radix-Stephanlae Tetrandrae Radix group (GS group), with 10 rats in each group. Rat model of wind-cold-dampness RA was induced through the injection with type Ⅱ collagen emulsion and wind-cold-dampness stimulation. After the establishment of the model, the blank control group, collage Ⅱ model group and wind-cold-dampness syndrome model group were given normal saline, and the corresponding liquid medicine was given to each administration group. In the experiment, the thickness of the left posterior metatarsal of rats was measured every 3 days, and the swelling degree of metatarsal was calculated. The arthritis index (AI) was evaluated on the 38th day of the experiment. The serum rheamatoid factor(RF) content of rats was detected by enzyme linked immunosorbent assay (ELISA). The expressions of MMP-3 and TIMP-1 in ankle joint were detected by Western blot. The expressions of MMP-3 and TIMP-1 mRNA in ankle joint were detected by real-time fluorescence quantitative PCR (Real-time PCR).Result: Compared with the blank group, the swelling degree, AI score, serum RF content, MMP-3 protein expression and MMP-3 mRNA expression in ankle joints of coll age Ⅱ model group and model wind-cold-dampness syndrome group were significantly increased (PPPPPPConclusion: For rheumatoid arthritis with wind-cold-dampness arthralgia, mild and warm traditional Chinese medicine (TCM) has a better effect than the combination of mild and cold TCM or mild TCM drugs. The experimental results are basically consistent with the principle of "treating cold diseases with hot medicine". The mechanism of the compatibility in treating rheumatoid arthritis due to wind-cold-dampness arthralgia may be related to the reduction of MMP-3, the increase of TIMP-1 expression and the reduction of articular cartilage damage.

Experimental study on decompensation model of rabbit corneal limbal stem cells induced by alkali burn / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To explore a new method to induce the animal model of rabbit partial limbal stem cell deficiency(LSCD).<p>METHODS: LSCD was induced through corneal alkali burn, C57 mice and New Zealand rabbits were used to establish the animal models. Corneal alkali burn manipulation was accomplished in experimental animals under general anesthesia combined with surface anesthesia in the operated eye. Specifically, mice(<i>n</i>=30)were used to induce complete LSCD model. In brief, the filter paper(diameter of 3mm)that immersed in 1mol/L potassium hydroxide solution was placed on the central corneal surface of the left eye for 30s, followed by washing with saline. In addition, rabbits(<i>n</i>=19)were utilized to establish the partial LSCD model. Briefly, the nictitating membrane(third eyelid)was resected, and the filter paper(diameter of 5mm)that immersed in 1mol/L potassium hydroxide solution was placed on the superior temporal peripheral corneal surface of the left for 30s, followed by washing with saline. After surgery, the model eyes were treated with 0.5% Levofloxacin Hydrochloride Eye Drops four times a day. In addition, the slit-lamp microscope was adopted for observation and photo-taking before burn, as well as at 1, 2, 4wk and 2mo after burn; meanwhile, complications such as corneal ulcer and perforation were recorded. 2mo after surgery, the corneal goblet cell distribution was detected with impression cytology, and the severity of LSCD was classified according to slit-lamp microscopic findings and corneal impression cytology. The animals were sacrificed 2mo after surgery, cornea and conjunctiva sections were made to observe angiogenesis and goblet cell distribution in cornea. Animals died accidentally were not counted into the total number, and the successful induction rates of complete LSCD and partial LSCD models were compared.<p>RESULTS: Six out of the 30 mice died accidentally, while 2 developed corneal perforation after burn, and the remaining 22 had developed complete LSCD only, yielding the successful induction rate of 92%. 2mo after burn, extensive angiogenesis distribution in the superficial and deep corneal stromal layers could be observed, and pathological sections revealed corneal angiogenesis. Seven out of the 19 rabbits died accidentally, while the remaining 12 had various degrees of LSCD only(partial LSCD, average involving 1.17±0.39 quadrants). Additionally, no corneal perforation was observed, and the successful induction rate was 100%. The result of Fisher's exact test <i>P</i> value is 0.543, without statistical difference. No goblet cells were observed in the normal corneal region, while goblet cells were observed in the LSCD region, with the average density of 58.60±12.58 cell/HP.<p>CONCLUSION: Central corneal alkali burn can induce complete LSCD; however, some animals will experience failure in model induction due to corneal ulcer and perforation, LSCD is generally serious and may be combined with angiogenesis in deep cornea. Alkali burn in superior temporal cornea can induce partial LSCD, which may be combined with relatively minor corneal lesion, and the corneal angiogenesis is located in the superficial layer.

Correlation between blood pressure stage and serum cardiovascular active peptide Salusin levels in pa-tients with essential hypertension / 心血管康复医学杂志

Objective :To explore correlation between stage cardiovascular active peptide salusin levels and class of essential hy-pertension (EH).Methods :A total of 145 EH patients were selected ,including 45 cases of hypertension stage 1 ,52 cases of stage 2 and 48 cases of stage 3- Another 50 healthy normotensive subjects undergoing physical examination simultaneously were enrolled as healthy control group .Levels of blood lipids ,serum salusin-α and salusin-β were measured and compared a-mong above groups .Results :Compared with healthy control group ,there was significant reduction in serum salusin-α level ,a-long with EH stage (1 ,2 ,3)rose ,there was significant reduction in serum salusin-α level [(2-40 ± 0-18)ng／ml vs.(1-17 ± 0-33) ng／ml vs.(0-82 ± 0-04)ng／ml vs.(0-60 ± 0-09)ng／ml , P＝0-001 all]in EH patients ;there was significant rise in serum sa-lusin-β level ,along with EH stage (1 ,2 ,3) rose ,there was significant rise in serum salusin-β level [(2-47 ± 0-28)ng／ml vs. (3-90 ± 0-26)ng／ml vs.(4-43 ± 0-09)ng／ml vs.(4-74 ± 0-10)ng／ml ,P＝0-001 all]in EH patients.Spearman correlation anal-ysis indicated that EH stage was significant inversely correlated with serum salusin-α level (r＝ －0-936 ,P＝0-001) ,and signif-icant positively correlated with serum salusin-β level (r＝0-526 , P＝0-001).Conclusion :Serum salusin-α and salusin-β possess certain predictive significance for hypertension stage ,which may provide new thinking for antihypertensive therapy .

Etiology spectrum of virus in acute upper respiratory infection in children from 2017 to 2018 in Qingdao / 中华疾病控制杂志

Objective To understand the prevalence and epidemiological characteristics of respiratory viruses in influenza-like illness in children during March 2017 to March 2018 in Qingdao. Methods A random selection of influenza surveillance cases (influenza-like illness, ILI) among children in Qingdao area was selected as the research object, and 359 cases were detected. Nasopharyngeal swabs were collected for multiple-fluorescence real-time reverse transcription-polymerase chain reaction nucleic acid detection to screen 9 kinds of respiratory viruses. Results Among the 359 Cases, 200 cases were positive for at least 1 kinds of viruses, and the positive rate was 55.71%(200/359). Among these 200 cases, the most positive numbers were influenza B Yamagata (IVB Yamagata) 29.50%(59/200), followed by enterovirus 15.00%(30/200), respiratory adenovirus (AdV) 13.50%(27/200), respiratory syncytial virus A (RSVA) 12.5%(25/200), influenza A H1N1(IVA H1N1) 10.00%(20/200), etc. 2 cases were 3 kinds of mixed viruses infected and 1 case was 4 kinds of mixed viruses infected. Conclusions Nine kinds of respiratory viruses are prevalent in Qingdao during March 2017 -March 2018. The main prevalence viruses contain influenza B Yamagata, enterovirus, respiratory adenovirus, respiratory syncytial virus A, influenza A H1N1. There is obvious seasonal distribution of influenza, respiratory syncytial virus, enterovirus, metapneumovirus. A mixed infection exists between 9 kinds of respiratory viruses, and mixed infection occurs in the month of the virus epidemic.

Effects of mitochondrial aldehyde dehydrogenase 2 on autophagy-associated proteins in neonatal rat myocardial fibroblasts cultured in high glucose / 南方医科大学学报

OBJECTIVE@#To investigate whether autophagy mediates the effects of aldehyde dehydrogenase 2 (ALDH2) on the proliferation of neonatal rat cardiac fibroblasts cultured in high glucose.@*METHODS@#Cardiac fibroblasts were isolated from neonatal (within 3 days) SD rats and subcultured. The fibroblasts of the third passage, after identification with immunofluorescence staining for vimentin, were treated with 5.5 mmol/L glucose (control group), 30 mmol/L glucose (high glucose group), or 30 mmol/L glucose in the presence of Alda-1 (an ALDH2 agonist), daidzin (an ALDH2 2 inhibitor), or both. Western blotting was employed to detect ALDH2, microtubule-associated protein 1 light chain 3B subunit (LC3B) and Beclin-1 in the cells, and a hydroxyproline detection kit was used for determining hydroxyproline content in cell culture medium; CCK- 8 kit was used for assessing the proliferation ability of the cardiac fibroblasts after the treatments.@*RESULTS@#Compared with the control cells, the cells exposed to high glucose exhibited obviously decreased expressions of ALDH2, Beclin-1 and LC3B and increased cell number and hydroxyproline content in the culture medium. Treatment of the high glucose-exposed cells with Alda-1 significantly increased Beclin-1, LC3B, and ALDH2 protein expressions and lowered the cell number and intracellular hydroxyproline content, whereas the application of daidzin resulted in reverse changes in the expressions of ALDH2, Beclin-1 and LC3B, viable cell number and intracellular hydroxyproline content in high glucose-exposed cells.@*CONCLUSIONS@#Mitochondrial ALDH2 inhibits the proliferation of neonatal rat cardiac fibroblasts induced by high glucose, and the effect is possibly mediated by the up-regulation of autophagy-related proteins Beclin-1 and LC3B.