OBJECTIVE: To observe the effects of Huayu Tongluo (transforming stasis and unblocking collaterals) moxibustion on learning-memory ability and hippocampal mammalian sterile 20-like kinase 1 (Mst1)/nuclear factor κB (NF-κB) p65 pathway related to inflammatory response in rats with vascular dementia (VD). METHODS: A total of 60 male Wistar rats of SPF grade were randomly divided into a sham operation group (12 rats) and a modeling group (48 rats). VD model was established by the method of modified bilateral common carotid artery permanent ligation in the modeling group. Thirty-six rats with successful modeling were randomly divided into a model group, a moxibustion group and a western medication group, with 12 rats in each group. Huayu Tongluo moxibustion was applied at "Dazhui" (GV14), "Baihui" (GV20) and "Shenting" (GV24) in the moxibustion group, 20 min each time, once a day, 7 day-intervention was as one course, and 1 day-interval was taken between two courses, for a total of 3 courses. In the western medication group, piracetam was given 0.72 mg/kg by intragastric administration, twice a day, the course of intervention was same as that of the moxibustion group. The learning-memory ability was detected by Morris water maze test; the morphology of hippocampal CA1 region was observed by HE staining; the mRNA expression of Mst1, M1 microglia markers CD86, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) was detected by real-time PCR; the levels of IL-6 and TNF-α in hippocampus were detected by ELISA; and the protein expression of Mst1 and NF-κB p65 in hippocampus was detected by Western blot in rats of each group. RESULTS: Compared with the sham operation group, the escape latency was prolonged in the model group (P<0.05); compared with the model group, the escape latency was shortened in the moxibustion group and the western medication group (P<0.05). The cells in the CA1 region of hippocampus were disordered, cell collapse and irregular nuclei could be observed in the model group; compared with the model group, the cell arrangement in the CA1 region of hippocampus was more regular, and the damage was improved in the moxibustion group and the western medication group. Compared with the sham operation group, the mRNA expression of Mst1, CD86, IL-6 and TNF-α, as well as the protein expression of Mst1, NF-κB p65 in hippocampus were increased in the model group (P<0.05). Compared with the model group, the mRNA expression of Mst1, CD86, IL-6 and TNF-α, as well as the protein expression of Mst1, NF-κB p65 in hippocampus were decreased in the moxibustion group and the western medication group (P<0.05). Compared with the sham operation group, the levels of IL-6 and TNF-α in hippocampus were increased in the model group (P<0.05). Compared with the model group, the levels of IL-6 and TNF-α in hippocampus were decreased in the moxibustion group and the western medication group (P<0.05). CONCLUSION Huayu Tongluo moxibustion can improve the learning-memory ability of VD rats, the mechanism may be related to regulating the activation of microglia through Mst1/NF-κB p65 pathway, reducing the release of pro-inflammatory factors i.e. IL-6 and TNF-α, so as to alleviating the damage of inflammatory factors in the hippocampus of VD rats.
Animals ; Male ; Rats ; Moxibustion ; Hippocampus/metabolism* ; Rats, Wistar ; Dementia, Vascular/genetics* ; Memory/drug effects* ; Humans ; Transcription Factor RelA/genetics* ; Learning ; Protein Serine-Threonine Kinases/genetics* ; Acupuncture Points ; Interleukin-6/genetics* ; Signal Transduction/drug effects* ; Drugs, Chinese Herbal

ObjectiveTo establish the identification method of Dalbergiae Odoriferae Lignum（DOL） and its counterfeits by nuclear magnetic resonance hydrogen spectrum（¹H-NMR） combined with multivariate statistical analysis. Method¹H-NMR spectra of DOL and its counterfeits were obtained by NMR， and the full composition information was established and transformed into a data matrix， and the detection conditions were as follows：taking dimethyl sulfoxide-d₆（DMSO-d₆） containing 0.03% tetramethylsilane（TMS） as the solvent， the constant temperature at 298 K（1 K=-272.15 ℃）， pulse interval of 1.00 s， spectrum width of 12 019.23 Hz， the scanning number of 16 times， and the sampling time of 1.08 s. Similarity examination and hierarchical cluster analysis（HCA） were performed on the data matrix of DOL and its counterfeits， and orthogonal partial least squares-discriminant analysis（OPLS-DA） was used to analyze the data matrix and identify the differential components between them. In the established OPLS-DA category variable value model， the category variable value of DOL was set as 1， and the category variable value of the counterfeits was set as 0， and the threshold was set as ±0.3， in order to identify the commercially available DOL. The OPLS-DA score plot was used to determine the types of counterfeits in commercially available DOL， and it was verified by thin layer chromatography（TLC）. ResultThe results of similarity analysis and HCA showed that there was a significant difference between DOL and its counterfeits. OPLS-DA found that the differential component between DOL and its counterfeits was trans-nerolidol. The established category variable value model could successfully identify the authenticity of the commercially available DOL. The results of the OPLS-DA score plot showed that there were heartwood of Dalbergia pinnata and D. cochinchinensis in the commercially available DOL， and were consistent with the TLC verification results. ConclusionThere is a phenomenon that heartwood of D. pinnata and D. cochinchinensis are sold as DOL in the market. ¹H-NMR combined with multivariate statistical analysis can effectively distinguish DOL and its counterfeits， which can provide a reference for the identification of them.

ObjectiveTo analyze the migrating components absorbed into blood of the aqueous extract of Euphorbia helioscopia， and to explore the pharmacodynamic material basis of the aqueous extract of E. helioscopia against chronic obstructive pulmonary disease（COPD）. MethodUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS） was used to detecte the migrating components absorbed into blood of rats after intragastric administration of aqueous extract of E. helioscopia. An Agilent RRHD SB-C₁₈ column（3 mm×100 mm， 1.8 μm） was used with 0.1% formic acid aqueous solution（A）-acetonitrile（B） as the mobile phase for gradient elution（0-15 min， 5%-30%B； 15-20 min， 30%-50%B； 20-30 min， 50%-95%B； 30-35 min， 95%-5%B）， and the detection wavelength of 190-800 nm， column temperature of 40 ℃， flow rate of 0.3 mL∙min^-1 and injection volume of 4 μL. The electrospray ionization（ESI） was used in positive and negative ion modes， and the detection range was m/z 50-1 250. Network pharmacology was used to screen out the key components and the key targets of COPD through the interaction analysis. Metascape database was used to predict the molecular function， biological process， cellular composition and signal pathways mainly involved in the anti-COPD effect of E. helioscopia. Molecular docking technique was used to determine the affinity of key targets with key components. ResultA total of 29 migrating components absorbed into blood of rats were identified after intragastric administration of aqueous extract of E. helioscopia， 9 of which were prototype components and 20 were metabolites. Network pharmacological analysis showed that luteolin， quercetin， apigenin， naringenin and helioscopinolide C were the key components of E. helioscopia against COPD， and vascular endothelial growth factor A（VEGFA）， albumin（ALB）， protein kinase B1（Akt1）， tumor necrosis factor（TNF） and interleukin-6（IL-6） were the key targets. Molecular docking results showed that one diterpene lactone（helioscopinolide C） and three flavonoids（naringenin， luteolin， apigenin） in the migrating components absorbed into blood all had strong binding activity to the key targets of E. helioscopia against COPD. ConclusionNaringenin， helioscopinolide C， luteolin and apigenin may be the main anti-COPD active substances of E. helioscopia.

More and more studies have found that intestinal flora is associated with connective tissue diseases. This review summarizes characteristics of intestinal flora and its mechanism of action in connective tissue diseases, and mainly elaborates correlations between intestinal flora imbalance and 3 common connective tissue diseases (lupus erythematosus, systemic scleroderma and Sjogren′s syndrome) , its possible mechanism of action and related hypothesis. Probiotics can regulate the intestinal flora imbalance, and serve as one of treatments for these common connective tissue diseases.

Objective To explore the effect of umbilical cord mesenchymal stem cells with positive human leukocyte antigen(HLA)-G on inducing the production of regulatory T cells(Treg) in vitro.Methods Umbilical cord mesenchymal stem cells were isolated from umbilical cord of neonates. PEGFP-N1-HLA-G plasmid was transfected into the human umbilical cord mesenchymal stem cells by liposome transfection, as PEGFP-N1-HLA-G group. PEGFP-N1 empty vector plasmid was transfected into the human umbilical cord mesenchymal stem cells, as PEGFP-N1 group. The human umbilical cord mesenchymal stem cells without empty vector under the same conditions were set as blank control group. Markers of the umbilical cord mesenchymal stem cells were detected using flow cytometry. The expression of HLA-G protein in each group of cells was identified by Western Blot. After mixed-culturing with CD4+T cells in peripheral blood of healthy subjects for 24 h and 48 h, the proportion of CD4+CD25+Foxp3+Treg in total T cells of each group was detected by flow cytometry. Results CD45, CD34 and HLA-DR presented negative expression on umbilical cord mesenchymal stem cells, while CD29, CD44 and CD105 presented positive expression. HLA-G protein could be expressed in the PEGFP-N1-HLA-G group, which had statistically significant difference compared with the blank control group and PEGFP-N1 group (both P<0.01). After PEGFP-N1-HLA-G group and CD4+T cells were mixed-cultured for 24 h and 48 h, CD4+CD25+Foxp3+Treg accounted for (15.3±1.9)% and (14.3±2.1)% of the total T cells respectively, both of which presented statistically significant difference compared with the blank control group and PEGFP-N1 group (all P<0.05). Conclusions Umbilical cord mesenchymal stem cells with HLA-G gene modified can effectively induce the production of CD4+CD25+Foxp3+Treg in vitro.

Objective To discuss CT and MRI performance and pathologic features about pseudomyxoma peritonei (PMP)con‐firmed by pathology which improves the PMP diagnosis level .Methods The CT and MRI informations about five cases with pseud‐omyxoma peritonei confirmed by abdominal puncture or surgery were analyzed retrospectively .Analyze the CT and MRI performance and clinical pathologic characteristics .Results In 5 cases ,one case of appendiceal mucous adenocarcinoma ,one case of acute appendi‐citis surgeried 2 years ago ,one case without any primary tumor ,one case of duodenal mucous adenocarcinoma ,one case of bilateral ovarian cancer with intra‐abdominal separated cystic lesion and/or peritoneal diffuse pie “frozen abdomen” change showed by CT , MRI ,and one case with pressure trace like scallops sample and multiple nodular around liver and spleen .Conclusion The lesion area and the shape of peritoneum pseudo myxoma can be showed by CT ,MRI scans .There is a certain special imaging feature .

Objective To investigate the mastery degree of GCP (Good Clinical Practice) in Guang’anmen Hospital of China Academy of Chinese Medical Sciences (hereinafter referred to as“Guang’anmen Hospital”). Methods Totally 338 clinical researchers, covering 18 clinical professional sections and medical clinical trial institutions, in Guang’anmen Hospital with both qualification of clinical trials and GCP certificate were investigated in 23th-27th June 2014. The investigation was in the written answer sheet form, including four noun explanations and 36 multiple choice questions, which were related to GCP. Results 338 researchers could accurately explain the concepts of 4 nouns, and 57 (16.86%) researchers could do all multiple choice questions correctly. 141 (41.72%) researchers chose 1 wrong answer and 89 (26.33%) researchers chose 2 wrong answers. 39 (11.54%) researchers failed to answer 3-4 questions. Only 12 (3.55%) researchers failed to answer 5 or more questions. Conclusion The mastery degree of clinical researchers with qualification of clinical trials and GCP certificate in Guang’anmen Hospital is satisfied. In the future, pertinence and diverse training should be strengthened, in order to improve the overall level of the mastery degree of GCP.

Objective Through detecting the expression of chemokine CCL21 and its receptor CCR7 in thyroiditis of NOD mice induced by excessive iodine and thyroglobulin (Tg) immunization,to explore the mechanism of lymphocytic infiltration.Methods Seven to eight weeks old female NOD (non-obese diabetic) mice were used,the mice were divided into four groups by random number table,7 mice for each group.Control group:intoke distilled water;,Tg group:intoke'distilled water,use pig Tg emulsified in Freund's complete adjuvant subcutaneous injection to immunization,0.10 mg Tg for each mouse at 8 weeks old,strengthened immunity at 11 and 15 weeks old (0.05 mg Tg for each mouse,and emulsified by Freund's incomplete adjuvant); iodine excess (HI) group:intoke 0.05％ NaI water; iodine excess combined with Tg (HI + Tg) group:intoke 0.05％ NaI water,Tg immunization method was same to Tg group; all mice were raised in specific pathogen free (SPF) environment.All mice were executed at 16 weeks old,and removed the thyroid tissue.Morphologic change in thyroid was observed by HE staining.Immunofluorescence (IF) staining was used to observe CD3,CD22,CCL21 and CCR7 expression in thyroid.Real-time PCR was used to detect CCL21 mRNA expression in thyroid.Results The morphologic change of thyroid was observed under the light microscope,in HI group,thyroid follicle distended,colloid accumulation,lymphocytes accumulate together and disorganization was seen; Tg group thyroid had scatted lymphocytes; HI + Tg group enhanced inflammation response and disorganization.IF staining showed that lymphocytes infiltrated in thyroid were almost CD3+ T cells,only few scatted CD22+ B cells; CCL21 and CCR7 were negative in control group,CCR7 positive cells were scatted in Tg group but no CCL21 expression,in HI and HI + Tg groups,we could see CCL21 strong positive expression in inflammation area and also had CCR7 positive cells there.CCL21 mRNA relative expression in thyroid:Tg group (3.20 ± 0.18),HI group (1.72 ± 0.31),and HI ± Tg group (9.94 ± 0.32) increased compared to control group (1.00 ± 0.17,all P ＜ 0.01).Conclusion CCL21 and its receptor CCR7 have improved lymphocytic infiltration in autoimmune thyroiditis

ObjectiveTo understand the status and requirement of the clinical researchers in Guang’anmen Hospital of China Academy of Chinese Medical Sciences (hereinafter referred to as Guang’anmen Hospital).MethodsSelf-filling questionnaire was used. All medical care personnel in the clinical specialty with the GCP qualification were set as respondents. The three aspects of recognition degree of clinical trials quality, initiative of participation and knowledge demand for GCP were investigated. Data were analyzed through SPSS11.5.Results 95.6% of clinical researchers considered that the course of clinical trial was legal;89.9% of clinical researchers believed that the quality of clinical trial was good. Most clinical researchers participated in the clinical trials actively (86.9%) and they considered that the trials were helpful for them to enhance research ability and to issue paper.Conclusion Clinical researchers are familiar with GCP knowledge and can participate in the clinical trials actively. They still expect to learn more.