[Banxia Xiexin Decoction inhibiting colitis-associated colorectal cancer infected with Fusobacterium nucleatum by regulating Wnt/ß-catenin pathway].

This paper investigates the intervention effect and mechanism of Banxia Xiexin Decoction(BXD) on colitis-associated colorectal cancer(CAC) infected with Fusobacterium nucleatum(Fn). C57BL/6 mice were randomly divided into a control group, Fn group, CAC group [azoxymethane(AOM)/dextran sulfate sodium salt(DSS)](AOM/DSS), model group, and BXD group. Except for the control and AOM/DSS groups, the mice in the other groups were orally administered with Fn suspension twice a week. The AOM/DSS group, model group, and BXD group were also injected with a single dose of 10 mg·kg~(-1) AOM combined with three cycles of 2.5% DSS taken intragastrically. The BXD group received oral administration of BXD starting from the second cycle until the end of the experiment. The general condition and weight changes of the mice were monitored during the experiment, and the disease activity index(DAI) was calculated. At the end of the experiment, the colon length and weight of the mice in each group were compared. Hematoxylin-eosin(HE) staining was used to observe the pathological changes in the colon tissue. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of interleukin(IL)-2, IL-4, and IL-6 inflammatory factors in the serum. Immunohistochemistry(IHC) was used to detect the expression of Ki67, E-cadherin, and ß-catenin in the colon tissue. Western blot was used to detect the protein content of Wnt3a, ß-catenin, E-cadherin, annexin A1, cyclin D1, and glycogen synthase kinase-3ß(GSK-3ß) in the colon tissue. The results showed that compared with the control group, the Fn group had no significant lesions. The mice in the AOM/DSS group and model group had decreased body weight, increased DAI scores, significantly increased colon weight, and significantly shortened colon length, with more significant lesions in the model group. At the same time, the colon histology of the model group showed more severe adenomas, inflammatory infiltration, and cellular dysplasia. The levels of IL-4 and IL-6 in the serum were significantly increased, while the IL-2 content was significantly decreased. The IHC results showed low expression of E-cadherin and high expression of Ki67 and ß-catenin in the model group, with a decreased protein content of E-cadherin and GSK-3ß and an increased protein content of Wnt3a, ß-catenin, annexin A1, and cyclin D1. After intervention with BXD, the body weight of the mice increased; the DAI score decreased; the colon length increased, and the tumor decreased. The histopathology showed reduced tumor proliferation and reduced inflammatory infiltration. The levels of IL-6 and IL-4 in the serum were significantly decreased, while the IL-2 content was increased. Meanwhile, the expression of E-cadherin was upregulated, and that of Ki67 and ß-catenin was downregulated. The protein content of E-cadherin and GSK-3ß increased, while that of Wnt3a, ß-catenin, annexin A1, and cyclin D1 decreased. In conclusion, BXD can inhibit CAC infected with Fn, and its potential mechanism may be related to the inhibition of Fn binding to E-cadherin, the decrease in annexin A1 protein level, and the regulation of the Wnt/ß-catenin pathway.

[Tongxie Yaofang regulates tumor-associated macrophage polarization in colorectal cancer under chronic stress].

This study aims to investigate the intervention effect and mechanism of Tongxie Yaofang in regulating tumor-associated macrophage polarization on colorectal cancer under chronic stress. BALB/C mice were randomized into blank, control, model, mifepristone, and low-, medium-, and high-dose Tongxie Yaofang groups. The other groups except the blank and model groups were subjected to chronic restraint stress and subcutaneous implantation of colon cancer cells for the modeling of colon cancer under stress. Du-ring this period, the body mass and tumor size of each group of mice were recorded. The degree of depression in mice was assessed by behavioral changes. Enzyme-linked immunosorbent assay was employed to determine the levels of cortisol(CORT), 5-hydroxytryptamine(5-HT), norepinephrine(NE), M1-associated inflammatory cytokines [interleukin(IL)-1ß, IL-12, and tumor necrosis factor(TNF)-α], and M2-associated inflammatory cytokines(IL-4 and IL-10) in the serum. The tumor growth of mice in each group was regularly monitored by in vivo imaging. The histopathological changes of tumors in each group of mice were observed by hematoxylin-eosin staining. The proportions of CD86 and CD206 in the tumor tissue were detected by flow cytometry and immunofluorescence staining. Western blot was employed to determine the protein levels of Janus kinase(JAK)1, JAK2, JAK3, signal transducer and activator of transcription(STAT)3, and STAT6 in the tumor tissue. The results showed that chronic stress increased the immobility time of mice, elevated the serum levels of CORT, IL-4, and IL-10, lowered the levels of 5-HT, NE, IL-1ß, IL-12, and TNF-α, and promoted the growth of subcutaneous tumors. The tumor cells in the tumor tissue grew actively, with obvious atypia and up-regulated protein levels of CD206, JAK1, JAK2, JAK3, STAT3, and STAT6, and down-regulated protein level of CD86. The treatment with Tongxie Yaofang shortened the immobility time of mice, lowered the serum levels of CORT, IL-4, and IL-10, elevated the serum levels of 5-HT, NE, IL-1ß, IL-12, and TNF-α, and inhibited the growth of subcutaneous tumors in mice. Moreover, the treatment caused different degrees of necrosis in the tumor tissues, down-regulated the protein levels of CD206, JAK1, JAK2, JAK3, STAT3, and STAT6, and up-regulated the protein level of CD86. In summary, Tongxie Yaofang can promote the transformation of M2 macrophages to M1 macrophages and change the tumor microenvironment under chronic stress to inhibit the development of colorectal cancer, which may be related to the JAK/STAT signaling pathway.

[Decursin affects proliferation, apoptosis, and migration of colorectal cancer cells through PI3K/Akt signaling pathway].

We investigated the effects of decursin on the proliferation, apoptosis, and migration of colorectal cancer HT29 and HCT116 cells through the phosphatidylinositol 3-kinase(PI3K)/serine-threonine kinase(Akt) pathway. Decursin(10, 30, 60, and 90 µmol·L~(-1)) was used to treat HT29 and HCT116 cells. The survival, colony formation ability, proliferation, apoptosis, wound hea-ling area, and migration of the HT29 and HCT116 cells exposed to decursin were examined by cell counting kit-8(CCK8), cloning formation experiments, Ki67 immunofluorescence staining, flow cytometry, wound healing assay, and Transwell assay, respectively. Western blot was employed to determine the expression levels of epithelial cadherin(E-cadherin), neural cadherin(N-cadherin), vimentin, B-cell lymphoma/leukemia-2(Bcl-2), Bcl-2-associated X protein(Bax), tumor suppressor protein p53, PI3K, and Akt. Compared with the control group, decursin significantly inhibited the proliferation and colony number and promoted the apoptosis of HT29 and HCT116 cells, and it significantly down-regulated the expression of Bcl-2 and up-regulated the expression of Bax. Decursin inhibited the wound healing and migration of the cells, significantly down-regulated the expression of N-cadherin and vimentin, and up-regulated the expression of E-cadherin. In addition, it significantly down-regulated the expression of PI3K and Akt and up-regulated that of p53. In summary, decursin may regulate epithelial-mesenchymal transition(EMT) via the PI3K/Akt signaling pathway, thereby affecting the proliferation, apoptosis, and migration of colorectal cancer cells.

Establishment and application of four long-term culture cell lines of the olive flounder Paralichthys olivaceus blastocysts.

Four new embryonic cell lines derived from blastocysts of the olive flounder Paralichthys olivaceus, an important commercial marine fish, were established and characterized. They were designated as PoEFCI, PoEFCII, PoEFCIII, and PoEFCIV and were all fibroblastic cells. The cells were cultured in DMEM/F-12 medium supplemented with antibiotics, FBS, and growth factors at temperature of 25 °C and subcultured for >100 passages over 18 months. The origin of the cell lines was confirmed by examining the partial sequences of the cytochrome oxidase c subunit I (COI) gene of the flounder mitochondrial DNA (mtDNA). The four cell lines showed different growth curve patterns. According to the results of gene and protein expression and enzyme activity, the cell lines PoEFCI, PoEFCII, and PoEFC III could be pluripotent. The cells of all four cell lines were also successfully transfected with the green fluorescent protein (GFP) reporter gene, suggesting that they could be used to study gene function in the flounder or other fish. More importantly, PoEFCI-III were sensitive to chromium (Cr) and red sea bream Pagrus major iridovirus (RSIV), so they could be used as a powerful tool for the study of the toxicological investigation of heavy metals and RSIV in fish. Therefore, these cell lines would be useful for biotechnological and toxicological research on marine fish as an in vitro biological system.

[Visualized analysis of knowledge map for traditional Chinese medicine prevention and treatment of pulmonary nodules based on CiteSpace].

This study aims to analyze the current status and development trend of the prevention and treatment of pulmonary nodules(PN) with traditional Chinese medicine(TCM) based on knowledge map and to provide both references and suggestions for future research directions. CNKI, Wanfang, VIP, and SinoMed were searched for relevant papers from the inception to December 31, 2021. Eligible articles were included according to the inclusion and exclusion criteria. The line chart was drawn based on the annual publication volume of articles, and the research interests of this field were learned. The knowledge maps of prevention and treatment of PN with TCM were drawn in CiteSpace 5.8.R1, and the authors, institutions, contents, and hotspots were analyzed. A total of 122 articles were included and the line chart demonstrated that the annual publication volume has been rising since 2018. According to the knowledge maps, the most prolific author was ZHANG Xiao-mei and there were four main research teams. Beijing University of Chinese Medicine and its affiliated hospitals were in a leading position in this field. The main research contents were disease, pathogenesis, and treatment, and the hotspots were data mining and TCM syndrome. The research on prevention and treatment of PN with TCM has become an increasing field of interest in recent years. In the future, the cross-regional cooperation and communication between research teams and institutions should be strengthened for more real-world studies and basic studies about the prevention and treatment of PN with TCM, so that the high-level evidence can be obtained and the underlying mechanisms of TCM formulae in the treatment of PN be clarified.

The Analgesic Benefits of Ketorolac to Local Anesthetic Wound Infiltration Is Statistically Significant But Clinically Unimportant: A Comprehensive Systematic Review and Meta-Analysis.

Objective: Even though ketorolac-infiltration is said to provide superior postoperative analgesic benefits in different surgical procedures, its safety and efficacy remain to be validated because of the lack of high-quality evidence. We aimed to summarize the efficacy and safety of ketorolac-infiltration based on published randomized-controlled trials (RCTs). Approach: This work followed Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, assessing the methodological quality of systematic reviews and the Cochrane Collaboration recommendations. We searched for RCTs evaluating the efficacy of ketorolac-infiltration in adults in the PubMed, Web of Science, Embase, Cochrane Library, Chinese databases, and Google Scholar. The two co-primary outcomes of this meta-analysis were rescue analgesic consumption in the 24-h postoperative period and rest pain scores. Results: Twelve trials (761 patients) were analyzed. Ketorolac-infiltration provided a clinically unimportant benefit in morphine consumption (mean difference, -2.81 mg; 95% confidence interval [CI], -5.11 to -0.50; p = 0.02; moderate-quality evidence). Low-to-moderate quality evidence supported a brief (2-6 h), clinically subtle, but statistically consistent effect of surgical site ketorolac-infiltration in reducing wound pain at rest. High-quality evidence supported shorter hospital stays for surgical patients receiving local ketorolac-infiltration when compared to controls (mean difference, -0.12 days; 95% CI, -0.17 to -0.08; p < 0.00001). Further, ketorolac-infiltration does not improve any opioid-related side effects. Innovation: Ketorolac-infiltration provides statistically significant but clinically unimportant benefits for improving postoperative wound pain. Conclusion: Overall, despite the fact that current moderate-to-high quality of evidence does not support routine using of ketorolac as an adjuvant to local anesthetic for wound infiltration, these findings underscore the importance of optimizing agents and sustained delivery parameters in postoperative local anesthetic practice. Clinical Trials.gov ID: CRD42021229095.

Hangeshashinto for preventing oral mucositis in patients receiving cancer treatment: protocol for a systematic review and meta-analysis.

INTRODUCTION: Hangeshashinto has been employed for oral mucositis prevention in patients receiving cancer treatment, but the evidence has not been sufficiently robust to guide clinical decision-making. This study will therefore be undertaken to assess the effectiveness of Hangeshashinto for preventing oral mucositis in patients with cancer who are receiving treatment. METHODS AND ANALYSIS: The databases will include PubMed, Embase, the Cochrane Library, Chinese databases and Japanese databases. The literature will be searched from the databases' inception until May 2021. Other sources, such as potential grey literature, reference lists from included studies and relevant systematic reviews and conference papers, will also be searched. The primary outcome is the incidence of mucositis of any severity, and the secondary outcomes are interruptions to cancer treatment, oral pain and nutritional status. The risk of bias of eligible studies will be assessed using the Cochrane Collaboration's 'risk of bias' tool. Both the Q test and I2 statistic will be performed to assess statistical heterogeneity. If I2 >50%, sensitivity and subgroup analysis will be conducted. The quality of evidence will be rated according to the Grading of Recommendations, Assessment, Development and Evaluation approach. Egger's test will be used to assess reporting bias. ETHICS AND DISSEMINATION: This systematic review will evaluate only published data; therefore, ethical approval is not required. PROSPERO REGISTRATION NUMBER: CRD42020216145.

External therapies of traditional Chinese medicine combined with sodium hyaluronate injected in articular cavity therapy on knee osteoarthritis: Meta-analysis / 中国中药杂志

To systematically evaluate the efficacy and safety of external therapies of traditional Chinese medicine(TCM) combined with sodium hyaluronate(SH) injected in articular cavity therapy on knee osteoarthritis(KOA). The following databases such as CNKI, WanFang, VIP, CBM, PubMed and Medline were researched to collect the randomized controlled trails on external therapies of TCM combined with sodium hyaluronate injected in articular cavity therapy on KOA. The selection of studies, assessment of methodological quality and data extraction were performed independently by two researchers. The methodological quality was assessed by using the Cochrane system evaluation methodology and Meta-analysis were performed by using Cochrane Collaboration's the RevMan 5.3 software. Forteen studies involving 1 449 patients were included. All of the trails were not adequate enough in methodological quality. Meta-analysis indicated that compared with control group, external therapies of TCM combined with sodium hyaluronate injected in articular cavity could raise effectiv rate(<0.000 01) and cure-rate(<0.000 01), improve Lysholm score(=0.003) and reduce VAS score(<0.000 1). But two groups have no difference in Womac score (=0.13).Compared with the treatment with sodium hyaluronate injected in articular cavity, external therapies of TCM combined with sodium hyaluronate injected in articular cavity, a promising treatment options, can be complementary advantages, improve the clinical curativ effect. But it still needs low risk and high quality clinical trials to verify.

The inhibitory effect of salvianolic acid B on TGF-ß1-induced proliferation and differentiation in lung fibroblasts.

Salvianolic acid B (Sal B), one of the major water-soluble compounds of Danshen (a popular Chinese herb), possesses many of the biological activities, such as antifibrogenic effect in liver and renal diseases. Transforming growth factor-ß1 (TGF-ß1) plays a central role in the development of pulmonary fibrosis by stimulating extracellular matrix (ECM) accumulation and activating fibroblasts. Here, we investigated the effects of Sal B on cell proliferation, collagen synthesis, endogenous TGF-ß1 production, and α-smooth muscle actin (α-SMA, a marker of myofibroblasts) expression in human lung fibroblasts stimulated by TGF-ß1 in vitro. The cell proliferation rates were analyzed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assay. The expression of TGF-ß1 and type I collagen at both the mRNA and protein levels was detected by reverse transcription polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and radioimmunoassay, respectively. The α-SMA expression was detected by Western blot. TGF-ß1 treatment of lung fibroblasts increased cell proliferation rates, and enhanced the expression level of type I collagen, endogenous TGF-ß1 production, and α-SMA expression (P < .05). The treatment with only Sal B did not affect the proliferation and differentiation of lung fibroblasts. Interestingly, Sal B was found to inhibit TGF-ß1-induced cell proliferation, expression of type I collagen, endogenous TGF-ß1 production, and α-SMA expression in lung fibroblasts. Moreover, the inhibitory effect of Sal B on TGF-ß1-induced proliferation and differentiation in lung fibroblasts was more significant when treated with high-dose Sal B (1 µmol/L versus 10 µmol/L, P < .05). These data demonstrate that Sal B inhibits TGF-ß1-induced cell proliferation and differentiation in vitro experiment.

[Boiling extraction of oleuropein at low temperature and reduced pressure].

OBJECTIVE: To look for optimum extraction techniques for oleuropein by boiling olive leaves at low temperature and reduced pressure. METHOD: According to single factor experiment (SFE) design, the effects of seven factors, the impact of seven factors, type of solvent, temperature, time, ratio of material to liquid, ethanol concentration, vacuum degree and extraction times, on extraction yield of oleuropein were investigated. Based on the results of SFE, four more important factors, temperature, time, ratio of material to liquid and ethanol concentration, were selected in L9 (3(4)) orthogonal experiment (OE) to compare with those extracted with traditional methods. RESULT: The optimum conditions for boiling extraction of oleuropein at low temperature and reduced pressure were as follows: temperature 60 degrees C, time 20 min, ratio of material to liquid 1:30 and ethanol concentration 85%. The conditions presented an extraction yield of 5.90%. CONCLUSION: Compared with traditional extraction methods and the ultrasound assisted extraction method, boiling extraction techniques at low temperature and reduced pressure were so quick and efficient that it has a good application prospect.

Study on preventive effect of jiangu granule on osteoporosis in ovariectomized rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the preventive effect of Jiangu granule (JGG) on experimental primary osteoporosis type I.</p><p><b>METHODS</b>Osteoporosis model was established through ovary resection of female rats. Bone mineral density (BMD) was measured with double energy X-ray absorptiometry. Level of endocrine markers, including osteocalcin (BGP), estradiol (E2) and calcitonin (CT) in serum were examined by RIA. And uterus parameters was calculated also.</p><p><b>RESULTS</b>JGG could significantly increase BMD and uterus index, improve the levels of E2 and CT in serum, at the same time reduce the BGP level.</p><p><b>CONCLUSION</b>JGG can efficiently prevent type I primary osteoporosis or delay its occurrence by enhancing the function of endocrine system, coordinating the action of calcium related hormone, reducing bone turnover rate and increasing BMD.</p>