Regulatory Effect and Mechanism of Yichang Sanjie Granules on Intestinal Flora and Immune Function in Mice with Colon Cancer / 广州中医药大学学报

Objective To observe the regulating effect and mechanism of Yichang Sanjie Granules on intestinal flora and immune function in mice with colon cancer.Methods Sixty mice were randomly divided into six groups,i.e.,the normal group,the model group,the low-,medium-and high-dose groups of Yichang Sanjie Granules,and the overexpression of melanoma absent gene 2(AIM2)plasmid(pcDNA-AIM2)intervention group,with 10 mice in each group.Colorectal cancer model was prepared by oxidized azomethine(AOM)/dextran sulfate sodium(DSS)induction method in all groups except normal group.After drug administration,the survival curves of mice in each group were plotted and the tumor volume was calculated;serum levels of immunoglobulin(Ig)G,IgM,interleukin(IL)-1β and IL-18 were detected by enzyme-linked immunosorbent assay(ELISA);peripheral blood levels of CD3+,CD4+,CD8+ T cells were detected by flow cytometry;the splenic index was determined;Hematoxylin-eosin(HE)staining was used to observe the pathological changes in colon tissues;16S-rDNA intestinal flora sequencing was used to detect the α-diversity of intestinal flora and the structure of intestinal flora communities;and protein immunoblotting(Wetsern Blot)was used to detect the protein expressions of AIM2,apoptosis-associated speckled-like protein containing a CARD(ASC),and cystatinase-1(caspase-1)in colon tissues.Results Compared with the normal group,the survival rate,serum levels of IgG and IgM,peripheral blood levels of CD3+ and CD4+ and CD4+/CD8+ ratio,protein expression levels of colon tissue AIM2,ASC and caspase-1 in the model group were significantly decreased,and the tumor volume,serum levels of IL-1β and IL-18,peripheral blood level of CD8+,and splenic index were significantly increased(all P＜0.05),and the HE staining results showed the characteristic manifestations of colon cancer;compared with the model group,the survival rate,serum levels of IgG and IgM,peripheral blood levels of CD3+ and CD4+ and CD4+/CD8+ ratio,protein expression levels of colon tissue AIM2,ASC and caspase-1 in the low-,medium-and high-dose groups of Yichang Sanjie Granules and the pcDNA-AIM2 group were significantly increased,and the tumor volume,serum levels of IL-1β and IL-18,level of peripheral blood CD8+,and splenic index were significantly decreased(all P＜0.05),and the HE staining results showed the manifestations of colon cancer were improved.Compared with the normal group,the Observed index,Chao1 index,Shannon index,the relative abundance of Bacteroidetes,Proteobacteria,Muribaculaceae,Lachnospiraceae-NK4A136group,and Ruminiclostridium in the model group were significantly decreased,while the relative abundance of Firmicutes,Actinobacteria,Patescibateria,Lactobacillus,Odoribacter,Alistipes,Ruminococcaceae-uncultured and Bacteroides was increased in the model group(P＜0.05);compared with the model group,the Observed index,Chao1 index,Shannon index,the relative abundance of Bacteroidetes,Proteobacteria,Muribaculaceae,Lachnospiraceae-NK4A136group and Ruminiclostridium were significantly increased,and the relative abundance of Firmicutes,Actinobacteria,Patescibateria,Lactobacillus,Odoribacter,Alistipes,Ruminococcaceae-uncultured and Bacteroides was decreased in the low-,medium-and high-dose groups of Yichang Sanjie Granules and the pcDNA-AIM2 group(all P＜0.05).Conclusion Yichang Sanjie Granules can increase autoimmunity and improve intestinal flora structure in mice with colon cancer,and its mechanism is related to the activation of AIM2 inflammatory vesicles.

Heixiaoyao Powder interferes with microglia polarization in AD model mice by regulating NOX2/ROS/NF-κB signaling pathway / 中国中药杂志

The effect and mechanism of Heixiaoyao Powder on the polarization of microglia(MG) in APP/PS1 double transgenic mice were explored based on NADPH oxidase 2(NOX2)/reactive oxygen species(ROS)/nuclear factor kappaB(NF-κB) signaling pathway. Fifty 4-month-old male APP/PS1 mice were randomly divided into a model group, an MCC950 group(10 mg·kg~(-1)), and low-, medium-, and high-dose Heixiaoyao Powder groups(6.45, 12.89, and 25.78 g·kg~(-1)). Thirty male C57BL/6J mice of the same age and strain were randomly divided into a blank group, a blank + intragastric intervention group, and a blank + intraperitoneal injection group. Drug intervention lasted 90 days. Morris water maze test was used to detect learning and cognitive ability. Nissl staining and transmission electron microscopy were used to observe the pathological morphology and ultrastructure of hippocampal neurons. Immunofluorescence was used to detect the positive expression of M1-type marker CD16/32~+/Iba-1~+, M2-type marker CD206~+/Iba-1~+ of MG and the expression of hippocampal ROS. The colorimetric method was used to detect the content of malondialdehyde(MDA) and superoxide dismutase(SOD) in the hippocampus. Enzyme linked immunosorbent assay(ELISA) was used to detect the levels of inflammatory factors, including interleukin-6(IL-6), interleukin-8(IL-8), and tumor necrosis factor-α(TNF-α), in the hippocampus. Western blot was used to detect the protein expression of β-amyloid protein(Aβ), Iba-1, CD16/32, CD206, NOX2, NF-κB, p-NF-κB, NF-κB inhibitor alpha(IκBα), and p-IKBα in the hippocampus. The results showed that as compared with the blank group, the model group showed prolonged target quadrant movement distance and escape latency(P<0.01), shortened target quadrant retention time and percentage(P<0.01), disorganized neuronal cells with swelling, nuclear disappearance or bias, reduced number of cells, dissolved or absent Nissl bodies, and a clear area in the cytoplasm, damaged and shrunk cell membrane with abnormal cell morphology, few organelles in the cytoplasm, reduced and swollen mitochondria, increased MG M1-type marker CD16/32~+/Iba-1~+(P<0.01), decreased M2-type marker CD206~+/Iba-1~+(P<0.01), increased ROS activity and MDA content(P<0.01), decreased SOD level(P<0.01), elevated inflammatory factors IL-6, IL-8, and TNF-α(P<0.01), up-regulated protein expression and phosphorylation of Aβ, CD16/32, Iba-1, NOX2, NF-κB, and IKBα(P<0.01), and down-regulated CD206(P<0.01). There was no statistically significant difference between the blank group, the blank + intragastric intervention group, and the blank + intraperitoneal injection group. After the intervention of Heixiaoyao Powder, the Heixiaoyao Powder groups showed shortened target quadrant movement distance and escape latency(P<0.01), prolonged target quadrant retention time and percentage(P<0.01), increased and neatly arranged cells with relieved swelling, increased Nissl bodies, regular cell morphology, and intact cell membrane, relieved swelling of mitochondria, slightly expanded endoplasmic reticulum, decreased CD16/32~+/Iba-1~+(P<0.05 or P<0.01), increased CD206~+/Iba-1~+(P<0.01), decreased ROS activity and MDA content(P<0.01), increased SOD level(P<0.01), decreased content of inflammatory factors IL-6, IL-8, and TNF-α(P<0.01), down-regulated protein expression and phosphorylation of Aβ, CD16/32, Iba-1, NOX2, NF-κB, and IKBα(P<0.01), and up-regulated CD206(P<0.01). In conclusion, Heixiaoyao Powder can alleviate neuronal damage and improve the learning and memory abilities of APP/PS1 mice. The mechanism of action may be related to the inhibition of NOX2/ROS/NF-κB signaling pathway, regulating the polarization of MG, increasing the expression of M2 type, inhibiting the expression of M1 type, and reducing the release of inflammatory factor.

Study on the relationship between smoking behavior and other unhealthy behaviors among middle school students in 4 cities of China / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the situation of smoking behavior among the students of middle school in Beijing, Hangzhou, Wuhan and Urumchi and to analyze the relationship between smoking behavior and several unhealthy behaviors together with psychological troubles to provide evidence in developing an early intervention plan.</p><p><b>METHODS</b>The National Health Education Institute (NHEI) of Chinese Center for Disease Control and Prevention (CDC) provided relevant data on all middle schools in the 4 cities and then U.S. CDC randomly sampled 100 common middle schools from them with a special sampling process. The core questionnaire developed by the experts from WHO and other countries was used in the survey among 9015 sampled students.</p><p><b>RESULTS</b>Among all the sampled students, 29.4% of them had ever attempted cigarettes smoking while 6.6% of them tried tobacco in the 30 days before survey, 27.0% of the students with smoking behavior began smoking at the age of 9 or younger, 31.8% had learned how to refuse smoking from school education. The students with smoking behavior were more likely to drink alcohol, use drugs, bully others, be injured, miss classes, and have some psychological troubles than those without smoking behavior.</p><p><b>CONCLUSION</b>There were increasing trends noticed on the incidence of attempt and smoking cigarettes. Smoking was closely related to other unhealthy behaviors and psychological troubles. Comprehensive education activities on "no-smoking" should be implemented as early as possible among adolescents, as well as to promote training on life skills.</p>

The relationship between test anxiety and personality, self-esteem in grade one senior high students / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the relationship between test anxiety and personality, self-esteem in grade one senior high school students.</p><p><b>METHODS</b>Totally 538 senior high school students of grade one were investigated by Test Anxiety Scale (TAS), Eysenck Personality Questionnaire (EPQ) and Self-Esteem Scale (SES) in a Senior High School in Shandong Province.</p><p><b>RESULTS</b>The prevalence of test anxiety among all the surveyed students was rated 65.2%. The Psychoticism (P) (51.60 +/- 9.66) or Neuroticism (N) (51.57 +/- 10.75) factor score of EPQ in students with test anxiety was significantly higher than that in students without test anxiety (48.07 +/- 8.62, 45.65 +/- 10.14) (P < 0.001), while the Extroversion or Introversion (E) score (50.76 +/- 11.09) was on the contrary (53.68 +/- 11.60) (P < 0.01). The total score of TAS was significantly positively related to the P (r = 0.14) and N (r = 0.36) factor score and significantly negatively related to the E factor score of EPQ (r = -0.15) (P < 0.001). The prevalence of test anxiety in introversive students (72.3%) was higher than that in extroversive students (53.2%) (P < 0.05), and that in students with unstable emotion (81.4%) and in students with apparent psychoticism (84.1%) were also higher than that in those with stable emotion (41.0%) and in those without psychoticism (57.7%) (P < 0.01). The total score of SES in students with test anxiety (29.12 +/- 4.41) was significantly lower than that in students without test anxiety (30.29 +/- 4.25) (P < 0.01). The total score of TAS was significantly negatively related to the total score of SES (r = -0.23) (P < 0.001).</p><p><b>CONCLUSION</b>Test anxiety should be related to the personality and self-esteem, and the prevalence of test anxiety in introversive, unstable emotional, apparent psychoticism or low self-esteem students should be higher.</p>

Polyketide Synthases Screening from Sponge-associated Culturable Microorganisms / 微生物学通报

PKS gene was screened by PCR from thirty strains of spone-associated bacteria including twenty-one actinomycetes isolated from Craniella anstrialiensis and nine bacillus isolated from Dysidea avara in the South China Sea.As a result,a 669 bp KS domain gene was successfully amplified from Bacillus C89.BLAST analysis showed that the KS domains were most closely related to the KS sequences of Bacillus subtilis subsp.subtilis str.168 with 96% similarity.Phylogenetic analysis demonstrated the KS domain belong to trans-AT KS domains.This study demonstrated the existence of PKS gene in bacteria associated with sponge Dysidea avara for the first time,and provided proof for the hypothesis that sponge-associated bacteria are perhaps the true producers of many novel bioactive compounds in sponge.Meanwhile,this study lays a basis for the microbial screening for polyketide compounds production.