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ObjectiveTo collect and screen records concerning the spleen and stomach diseases and syndromes in ancient books of traditional Chinese medicine （TCM） using an automated framework and to systematically explore the concept evolution of spleen and stomach diseases and syndromes using the visualization method. MethodA total of 1 224 kinds of ancient book data in the Ancient Books of Traditional Chinese Medicine Database （V2.1） were analyzed using the automated testing tool Selenium WebDriver with the Python 3.8 programming language and the etree library of Lxml for automatic collection and statistics of the "book title" "author" "classification" "dynasty" "completion time"， and "informative abstract". After being checked and collated， the collected data were visually analyzed with Tableau （V2020.1.3） for figuring out the concept evolution of spleen and stomach diseases and syndromes in the past dynasties from the perspectives of symptoms and signs， etiology and pathogenesis， principle-method-recipe-medicinal， and prognosis. ResultA total of 7 203 clauses were automatically collected from 989 ancient books. It was found that in the pre-Qin period， there were few ancient books related to the spleen and stomach diseases and syndromes， and the understandings were confined to the superficial symptoms or signs and the basic etiology and pathogenesis. From the Han to Sui and Tang dynasties， the related concepts gradually increased and the descriptions about the manifestations are more detailed than those in previous dynasties. The etiology， diagnosis， and treatment system of the spleen and stomach diseases and syndromes were further perfected. In the Song， Jin， and Yuan dynasties， such concepts as independent signs，symptoms， as well as nature and location of spleen and stomach diseases and syndromes were enriched. In the Ming and Qing dynasties， a TCM syndrome differentiation and treatment system for spleen and stomach diseases and syndromes was formed， and the related concepts were gradually simplified and unified. ConclusionThe concepts of spleen and stomach diseases and syndromes have undergone an evolution from simplicity to complexity and then back to simplicity. There are numerous ancient books discussing the concepts of spleen and stomach diseases and syndromes， exhibiting a fluctuating yet rising trend with time. The automated framework enables the construction of a lightweight database of spleen and stomach diseases and syndromes. Based on data visualization， the concept evolution of the spleen and stomach diseases and syndromes from ancient times to the present has been efficiently uncovered， which is conducive to tracing the origin and development of spleen and stomach diseases and syndromes in TCM. This has provided reference for related research of spleen and stomach diseases in modern Chinese medicine.
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Objective:To explore the molecular mechanism of spleen-strengthening traditional Chinese medicine （TCM） Weichang'an granule in inhibiting the invasion and metastasis of human gastric cancer MKN45 cells. Method:MKN45 cells were cultured <italic>in vitro</italic> and incubated with different concentrations（600， 900， 1 200， 1 500 mg·L<sup>-1</sup>）of Weichang'an granule for 24， 48， 72 h. Cell counting kit-8 （CCK-8） assay was used to detect its effect on the cell proliferation. Western blot was used to detect the expression of RUN and FYVE domain containing 3（RUFY3） in normal gastric mucosa cells and different gastric cancer cell lines. The expression of RUFY3 in the gastric cancer cells after Weichang'an granule intervention （600， 900， 1 200 mg·L<sup>-1</sup>） was detected by Western blot. Lentivirus transfection technique was used to achieve the stable and silenced expression of RUFY3 in gastric cancer MKN45 cells. Transwell assay was used to evaluate the influence of Weichang'an granule and silenced RUFY3 on the metastasis and invasion ability of MKN45. E-cadherin，N-cadherin，Vimentin，Zinc-finger transcription factor （SNAIL1 and SNAIL2） protein expression levels were detected by Western blot. Result:RUFY3 expression in human gastric cancer cells was significantly higher than that in normal gastric mucosa．The protein expression of RUFY3 in MKN45 cells of silenced RUFY3 group was significantly lower than that in Lentivirus negative group （<italic>P</italic><0.01）． Weichang'an granule inhibited the expression of RUFY3 in human MKN45 gastric cancer cells （<italic>P</italic><0.05， <italic>P</italic><0.01） in a concentration-dependent manner. As compared with the blank group， both Weichang'an granule and silenced RUFY3 inhibited the metastasis and invasion ability of MKN45 （<italic>P</italic><0.01）. After Weichang'an granule and silenced RUFY3 treatment， the protein expression of epithelial marker gene E-cadherin was up-regulated （<italic>P</italic><0.01）， the protein expression of Vimentin and N-cadherin decreased， but with no statistical difference，while SNAIL1 and SNAIL2 were both significantly down-regulated （<italic>P</italic><0.01）. Conclusion:By targeting RUFY3 to regulate epithelial mesenchymal transformation， the spleen-strengthening TCM compound Weichang'an granule can inhibit the invasion and metastasis of gastric cancer.
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<b>Objective::To explore the effect of Weichang' an (WCA) on the autophagy of human gastric cancer MKN45 cells and its possible anti-cancer mechanism. <b>Method::MKN45 cells were cultured <italic>in vitro</italic> and incubated with different concentrations (250, 500, 1 000, 2 000 mg·L<sup>-1</sup>) of WCA for 24, 48, 72 h. Cell counting kit-8 (CCK-8) assay was used to detect the cell proliferation. AO/EB Dyeing (AO) staining and monodansylcadaverin (MDC) staining were used to observe the changes of the effect of WCA on autophagosome and autophagic vesicles in gastric cancer MKN45 cells at 48 h. Real-time polymerase chain reaction (Real-time PCR) and Western blot were used to detect microtubule-associated protein 1 light chain 3 (LC3), Beclin1, sequestosome 1 (p62), human autophagy-related gene 5 (ATG5), human autophagy-related gene 7 (ATG7) mRNA and protein expression levels. <b>Result::WCA showed a dose-and-time-dependent growth inhibition at the concentration above 1 000 mg·L<sup>-1</sup>. Compared with the blank group, WCA (500, 1 000, 2 000 mg·L<sup>-1</sup>) significantly inhibited the proliferation of MKN45 cells (<italic>P</italic><0.05, <italic>P</italic><0.01). AO staining and MDC staining showed that autophagosomes and autophagic sacs increased with the rise of WCA concentration compared with the blank group. Real-time PCR and Western blot showed that the expressions of autophagy-related proteins LC3-Ⅱ, Beclin1, ATG5, ATG7, mRNA and protein increased gradually after WCA (1 000 mg·L<sup>-1</sup>) intervention, while p62 expression decreased (<italic>P</italic><0.05, <italic>P</italic><0.01). <b>Conclusion::WCA induces the autophagy of human gastric cancer MKN45 cells in a time-and dose-dependent manner <italic>in vitro</italic>, which may be related to the up-regulation of LC3-Ⅱ, Beclin1, ATG5 and ATG7 as well as the down-regulation of p62 and LC3-Ⅰ.
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Objective: To observe the effect of Weichang'an on the tumor growth and lymph node metastasis, and RUN and FYVE domain-containing protein 3(RUFY3),Zinc finger protein 1(SNAI1),vascular endothelial growth factor(VEGF),epithelial-mesenchymal transition(EMT)-related proteins in nude mice with subcutaneous xenograft tumor human gastric carcinoma MKN45, so as to discuss the mechanism of Weichang'an on MKN45 human gastric metastasis. Method: The nude mice model of human gastric carcinoma MKN45 cells was established and randomly divided into normal saline group (0.5 mL/mouse), Weichang'an granule group (3.54 g·kg-1) and Weichang'an decoction group (35.49 g·kg-1). The tumor weight and volume of axillary lymph nodes in each group were observed. The morphology of lymph nodes in each group was detected by hematoxylin-eosin (HE) staining. The expressions of related proteins were detected by immunohistochemistry, including RUFY3,SNAI1,VEGF,E-cadherin,N-cadherin,Vimentin. Result: Compared with the normal saline group, the tumor weight and volume of axillary lymph node were decreased (PPPConclusion: Both Weichang'an granule and Weichang'an decoction can inhibit the tumor growth and metastasis of axillary lymph nodes, obviously down-regulate the expressions of RUFY3,SNAI1,VEGF,N-cadherin,Vimentin and up-regulate the expression of E-cadherin in human gastric MKN45 subcutaneous transplanted tumor in nude mice. This suggested that Weichang'an may inhibit the tumor metastasis through regulation expressions of RUFY3,SNAI1,VEGF and EMT-related proteins.
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OBJECTIVE@#To develop an improved version of the Quality-of-Life Assessment instrument for Lung Cancer Patients Based on Traditional Chinese Medicine (QLASTCM-Lu) and to evaluate its psychometric property.@*METHODS@#The structured group method and the theory in developing rating scale were employed to revise the preliminary scale. The psychometric property (reliability, validity, and responsiveness) of the established QLASTCM-Lu (modified) were evaluated by quality of life data measured in 100 lung cancer patients. Statistical analyses were made accordingly by way of correlation analysis, factor analysis and paired t-test.@*RESULTS@#The internal consistency reliability of the overall scale and all domains was from 0.80 to 0.94. Correlation and factor analyses demonstrated that the scale was good in construct validity. The criterion validity was formed with European Organization for Research and Treatment of Cancer-Quality of Life Questionnaire-Lung Cancer (EORTC QLQ-LC43) as the criterion. Statistically significant changes were found apart from such domain as "mental condition" and "social function", with the standardized response means being close to those of QLQ-LC43.@*CONCLUSION@#QLASTCM-Lu (modified) could be used to measure the quality of life of lung cancer patients with good reliability, validity and a certain degree of responsiveness.
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<p><b>OBJECTIVE</b>Chinese jianpi herbal recipe Weichangan (WCA) could increase the survival rate of advanced gastric cancer. This study was designed to investigate the molecular mechanism of WCA in treatment of gastric cancer by cDNA array, real-time quantitative PCR and immunohistochemical technique.</p><p><b>METHOD</b>A human gastric adenocarcinoma cell line SGC-7901 grafted onto nude mouse was used as the animal model. The mice were divided into 3 groups, one control and the two representing experimental conditions. Animals in the two experimental groups received either WCA over a 34-day period or 5-fluorouracil (5-FU) over 6-day period starting at 8th day after grafting. Control animals received saline on an identical schedule. Animals were killed 41 days after being grafted. To assess the effect of therapy tumor weight was determined by a electron balance immediately after the animals killed. SP immunohistochemical method was used to detect the expression of proliferating cell nuclear antigen (PCNA) in xenografts. For detection of apoptotic cells, apoptotic indices (AI) were examined by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL) method. SP method was also used to detect the expression of cleaved Caspase-3. The expression profiles in paired WCA treated gastric cancer samples and the N. S. control samples were studied by using a cDNA array representing 14, 181 cDNA clusters. The alterations in gene expression levels were confirmed by real-time quantitative PCR. SP method was used to detect the expression of Phospho-Stat3 (Tyr705) and bcl-2.</p><p><b>RESULT</b>When compared with controls, tumor growth was significantly inhibited by treatment with the WCA or 5-FU (P < 0.01, respectively). The average of tumor inhibitory rate in WCA group was (44.32 +/- 5.67)% and 5-FU (47.04 +/- 11.33)%. The average labeling index (LI) for PCNA in WCA group and 5-FU group was significantly decreased compared with the control group respectively. AI of human gastric cancer xenografts in nude mice was significantly increased to (9.72 +/- 4.51)% using TUNEL method in WCA group compared with the controls (2.45 +/- 1.37)%. 5-FU group was also found a significantly increased AI compared with the controls. The expression of cleaved Caspase-3 in WCA group and 5-FU group was significantly increased compared with the control group respectively. There were 45 different expression ESTs among the control sample pool and WCA sample pool. There were 24 ESTs up-regulated in WCA samples and 21 ESTs down-regulated. These 45 ESTs contains 35 cloned genes and 11 unknown ESTs. By using Real-time Quantitative PCR, the expression level of Stat3 (2(-deltadeltaCT) = 0.16) , RIPX (2(-deltadeltaCT) = 0.18), ROD1 (2(-deltadeltaCT) = 0.23) and bcl-2 (2 (-deltadeltaCT) = 0.10) was lower in WCA group than that in control group respectively. The expression of Phospho-Stat3 (Tyr705) and bcl-2 in WCA group and 5-FU group was significantly decreased compared with the control group respectively.</p><p><b>CONCLUSION</b>Chinese jianpi herbal recipe WCA could inhibit gastric cancer cell SGC-7901 growth in vivo. WCA could induce gastric cancer cell apoptosis and suppress proliferation. Its mechanisms might be involved in the down-regulation of Stat3, RIPX, ROD1 and bcl-2 gene.</p>