To study the absorption and transepithelial transport of six coumarins (umbelliferone, osthole, columbianadin, columbianetin acetate, angelol-A and angelol-B, isolated from the roots of Angelica pubescens f. biserrata) in the human Caco-2 cell monolayer model.

In order to clarify the chemical composition of Tongmai Keli, a HPLC fingerprint was established, and the 22 peaks were characterized by LC-DAD-MS. The herbal sources of these peaks were assigned. The results implied that genistin 8-C-glucoside, puerarin, daidzein 8-C-apiosyl (1, 6)-O-glucoside, daidzin, and salvianolic acid B were the main constituents of in Tongmai Keli. Ten batches of Tongmai Keli produced by different pharmaceutical companies were analyzed. Although different batches contained similar compounds, the contents of major compounds were significantly different. The method established in this study could be used for the quality control of Tongmai Keli.

Objective To observe the effects of different doses of Sappan Lignum and Chuanxiong Rhizoma on tumor stem cells marker ABCG2 in vivo. Methods Sphere cells obtained from serum culture were inoculated in nude mouse armpit, which were randomly divided into 5 groups: control group, Sappan Lignum high- and low-dose groups, and Chuanxiong Rhizoma high- and low-dose groups. Each medication group was given relevant medicine for gavage. 21 days later, inhibition tumor rate and ABCG2 protein and mRNA expression were detected with confocal microscope, Western blot, and RT-PCR. Results The sphere cells obtained from serum free culture had the abilities of cancer stem cells, such as proliferation, anti-aptosis and high expression of cancer stem cells markers. Chuanxiong Rhizoma high- and low-dose groups could inhibit tumor growth (P<0.05), and the inhibitory rate of Chuanxiong Rhizoma low-dose group was higher than the Chuanxiong Rhizoma high-dose group. Sappan Lignum high- and low-dose groups inhibited tumor growth without statistical significiance (P>0.05). Compared with the control group, Chuanxiong Rhizoma low-dose group could significantly inhibit the expression of resistant protein of ABCG2. Sappan Lignum high- and low-dose groups could not inhibit the protein expression of ABCG2. Each medication group up-regulated the mRNA expression of ABCG2 except for Chuanxiong Rhizoma low-dose group. Conclusion Low dose of Chuanxiong Rhizoma can inhibit the expression of ABCG2 protein levels, which can be the targeting killer for cancer stem cells.

Objective To detect the genomic copy number variations(CNVs) of mice embryonic neural tissue with neural tube defects (NTDs) induced by methotrexate (MTX),and investigate the molecular genetic mechanisms between folic acid metabolism disorders and NTDs pathogenesis.Methods C57BL/6J NTD mice model was induced by MTX on gestational day 7.5,and the maternal serum and NTD embryonic neural tissues were collected;the array-comparative genomic hybridization(array-CGH) assay was utilized to analyze the whole genomic CNVs in NTD embryonic neural tissues;reverse transcription polymerase chain reaction(RT-PCR) was used to confirm the positive results;the maternal serum concentrations of folic acid and related metabolites and dihydrofolate reductase (DHFR) activity were detected by liquid chromatography-tandem mass spectrometry and enzymatic methods,respectively.Results ArrayCGH and RT-PCR results showed the 3 high confidence CNVs on XqE3,XqA1.1-qA2 and XqA1.1 in the NTD embryonic neural tissues.The NTD maternal serum concentrations of 5-methyltetrahydrofolate,5-formyltetrahydrofolic acid,S-adenosyl methionine and DHFR activity were reduced significantly compared with the control group,and there were statistical differences(all P ＜0.05).Conclusions There are obvious CNVs in embryonic neural tissue of NTD mice induced by MTX and folic acid dysmetabolism might cause mice embryonic neural tube developmental disorders through CNVs.

Objective:To understand the epidemic characteristics and influential factors of brucellosis patients in Laizhou, Shandong Province, and to provide scientific basis for prevention and control of brucellosis.Methods:Retrospective analysis was conducted to collect the surveillance data and epidemiological investigation results of confirmed cases of brucellosis in Laizhou from 2016 to 2019 through the "Infectious Disease Report Information Management System of the China Disease Prevention and Control Information System". The general situation, epidemiological characteristics, and clinical characteristics of brucellosis were analyzed by descriptive epidemiology. Using the case-control study method, 81 confirmed cases of brucellosis and 162 healthy people in the same area, same gender and same age as confirmed cases were selected to analyze the influential factors of brucellosis.Results:Totally 222 cases of brucellosis had been reported in Laizhou, with an average annual incidence of 6.57/100 000. The onset age of brucellosis was mainly from 45 to 59 years old, accounting for 46.40% (103/222); male-to-female ratio was 2.42 ∶ 1.00 (157 ∶ 65); the occupation distribution was mainly farmer, accounting for 95.95% (213/222). The onset time of brucellosis was mainly from March to July, accounting for 58.11% (129/222). Cases of brucellosis were reported in 17 townships (streets) of Laizhou, and the top five townships were Zhuqiao, Shahe, Tushan, Chengguo and Yidao towns in order of incidence. Among 222 cases of brucellosis, the shortest interval from onset to diagnosis was 5 d, the longest was 36 d, and the median was 11 d. The main clinical manifestations of the patients were fever (89.19%, 198/222), fatigue (80.63%, 179/222), joint muscle soreness (72.97%, 162/222), and a few accompanied by liver, spleen and testis enlargement. The logistic regression analysis showed that contact with urine, feces and other excrements of diseased animals [odds ratio ( OR) = 6.244, 95% confidence interval ( CI): 1.609 - 24.231] and family members suffering from brucellosis ( OR = 16.363, 95% CI: 2.250 - 119.023) were risk factors of brucellosis. Conclusions:The incidence of brucellosis is mainly among 45 - 59 middle-aged farmers in Laizhou and the incidence peaks in spring and summer. Direct contact with excrements of diseased animals and family members suffering from brucellosis are the risk factors of brucellosis. Health departments should carry out health education and behavioral intervention for farmers in the season and areas with high incidence of brucellosis, so as to improve personal awareness of protection.

ObjectiveTo observe the distribution of traditional Chinese medicine （TCM） syndrome types in advanced pancreatic cancer patients， and explore the association between median survival time and different TCM syndromes and different intervention times of Chinese herbal medicine （CHM）. MethodsThe clinical data of 136 advanced pancreatic cancer patients who have received CHM for more than 3 months were collected retrospectively， including gender， age， family history， smoking history， drinking history， location of disease， lymph node metastasis， multiple distant metastasis， western medicine treatment methods， TCM diagnosis and treatment information， and survival time. The Kaplan-Meier （KM） estimator was used， and the median survival time of patients was calculated. The TCM syndrome type of each patient was judged， and the main single syndrome types and compound syndrome types were summarized. The median survival time was compared among different compound syndrome types. The patients were further divided into the group of those having received CHM ≥6 months and those having received CHM ＜6 months. Whether receiving CHM ≥6 months was taken as the grouping variable， while the matching variables were age， gender， family history， smoking history， drinking history， location of disease， lymph node metastasis， multiple distant metastasis， surgery， chemotherapy， and radiotherapy when propensity score matching was performed， and the difference in median survival time between the two groups of patients before and after matching was compared. ResultsFor 136 cases of advanced pancreatic cancer， the top five single syndromes were spleen qi deficiency， liver blood stasis， liver qi stagnation， spleen dampness， and liver heat. The main compound types were liver constraint， spleen deficiency and blood stasis syndrome， liver-gallbladder damp-heat and blood stasis syndrome， liver constraint， qi stagnation and spleen deficiency syndrome， spleen-stomach yang deficiency and blood stasis syndrome， and spleen deficiency and dampness-heat internal accumulation syndrome. The overall median survival time before and after matching was 12.47 （7.70，17.10） months and 13.77 （8.83，17.20） months， respectively， and was significantly higher in the group treated with CHM ≥ 6 months than that treated with CHM ＜6 months （P＜0.05）. Among the 136 patients before matching， the median survival time of patients with spleen deficiency and dampness-heat internal accumulation syndrome was longest ［16.23 （14.17，19.40） months］， while that of patients with spleen-stomach yang deficiency and blood stasis syndrome was the shortest ［7.33 （5.80，12.83） months］. For patients with liver constraint， spleen deficiency and blood stasis syndrome， liver-gallbladder damp-heat and blood stasis syndrome， and spleen-stomach yang deficiency and blood stasis syndrome， those having received CHM ≥ 6 months have much longer median survival time than those having received CHM ＜6 months （P＜0.05）. Among the 108 patients after matching， the median survival time of those with spleen deficiency and dampness-heat internal accumulation syndrome was the longest ［15.23 （7.67，18.27） months］， while that of spleen-stomach yang deficiency and blood stasis syndrome was the shortest ［8.80 （6.90，16.17） months］. For patients with liver-gallbladder dampness-heat and blood stasis syndrome and spleen-stomach yang deficiency and blood stasis syndrome， the median survival time was higher in the group treated with CHM ≥ 6 months treated with CHM ＜6 months （P＜0.05）. ConclusionAfter treatment with CHM， advanced pancreatic cancer patients with spleen deficiency and damp-heat internal accumulation had a better prognosis， while those with spleen-stomach yang deficiency and blood stasis had a worse prognosis. Treatment with CHM ≥ 6 months could extend the median survival of advanced pancreatic cancer patients with liver-gallbladder damp-heat and blood stasis syndrome and spleen-stomach yang deficiency and blood stasis syndrome.