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Objective:To evaluate the clinical effect of treatment of activating blood and removing blood stasis, invigorating the spleen and soothing the liver for the patients with gastric collateral stasis syndrome and chronic atrophic gastritis (CAG).Methods:Randomized controlled trial. A total of 68 CAG patients admitted to the Huairou Hospital of Traditional Chinese Medicine from January 2018 to January 2021 who met the selection criteria were divided into 2 groups according to the random number table method, with 34 in each group. The control group received conventional western medicine treatment, such as inhibition of acid, protecting the gastric mucosa, and the observation group was treated with Traditional Chinese Medicine (TCM) herbal prescription of activating blood and removing blood stasis, invigorating the spleen and soothing the liver. Both groups were treated for 12 weeks. TCM symptom scores were performed before and after treatment. The serum level of pepsinogen Ⅰ(PG Ⅰ), pepsinogen Ⅱ (PGⅡ) were detected by ELISA, and the PG Ⅰ/PG Ⅱ ratio was calculated. Gastroscopic biopsy was performed to observe the changes of intestinal metaplasia of gastric mucosa and glandular atrophy, and to evaluate the clinical efficacy.Results:The total responsive rate was 85.3% (29/34) in the study group and 58.8% (20/34) in the control group. There was significant difference between the two groups ( χ2=9.35, P=0.030). After treatment, the scores of stomachache, fullness of feeling in the observation group were significantly lower than those in the control group ( t=2.97, 3.80, P<0.05). After treatment, the level of serum PG Ⅰ[(76.21 ± 17.35) mg/L vs. (66.8 ± 18.77) mg/L, t=2.15] and PG Ⅰ/PG Ⅱ [(4.67 ± 0.99) vs. (3.90± 1.25), t=2.81] in the study group were significantly higher than those in the control group ( P<0.05), and PG Ⅱ [(16.36 ± 1.85) mg/L vs. (17.42 ± 2.05) mg/L, t=2.24] was significantly lower than that of the control group ( P<0.05). After treatment, intestinal metaplasia and glandular atrophy was significantly more improved or reversed than those in the control group ( χ2=20.67,9.33, P<0.05). Conclusion:The methods of activating blood and removing blood stasis, invigorating the spleen and soothing the liver can reverse the precancerous lesions of patients with gastric collateral stasis syndrome of chronic atrophic gastritis and have a good prognosis.
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Objective To determine the effect of rapamycin(Rapa) on JAK2, ABCA3, and the immune checkpoint PD-1/PD-L1 in exosomes derived from JAK2 V617F positive HEL cells. Methods Human erythroleukemia HEL cells (JAK2 V617F mutation-positive) were cultured in vitro, and rapamycin was added at concentrations of 10, 50, and 100 nmol/L. The control group was established and cell proliferation inhibition rate was detected by CCK-8. Based on the inhibition rate of cell proliferation, cells intervened with 10 and 50 nmol/L Rapa were selected. Exosomes were extracted using a kit and identified by Western blot and flow cytometry. JAK2, ABCA3, and PD-1/PD-L1 mRNA changes in exosomes were detected by fluorescent quantitative PCR. The expression of exosome PD-1/PD-L1 protein was determined by flow cytometry. Results The exosomes extracted with the exosome kit all expressed characteristic CD9, CD63, and CD81 proteins, which were consistent with the general characteristics of exosomes. JAK2 mRNA can be detected in exosomes from HEL cells; Rapa reduced exosome production by HEL cells, and dose-dependently decreased gene expression of JAK2, ABCA3, and PD-L1 in exosomes. In addition, Rapa inhibited exosomal PD-L1 protein expression and had no significant effect on PD-1 protein expression. Conclusion HEL cells may transmit JAK2 mutant gene signals via exosomes. Rapa can reduce the production of exosomes and inhibits JAK2 mutated signals delivered by exosomes and suppresses PD-L1 protein expression.
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Aim To reveal the underlying mechanisms of the co-occurrence of ASXLI and JAK2ymr mutation by using human leukemia cell line HEL that carried homozygous /4A2V617F mutation in the elucidation of the role of ASXLI loss of function mutation in myeloproliferative neoplasms, so as to provide an important model for investigating the role of ASXLI mutation in myeloproliferative neoplasms at the cellular level. Methods HEL cell line with ASXLI knockout ( HEL-AKO) was established by using CRISPR/Cas9-mediated gene editing. And a series experiments based were preformed to verify the effect of ASXLI on HEL cell proliferation, clone formation and chemosensitivity. Results HEL-AKO cell line was successfully established, confirmed by sequencing results. We found that the loss of ASXLI could inhibit proliferation and induce cell cycle arrest at the G2/M phase. The colony-form- ing capacity of HEL-AKO cells was also markedly inhibited. Moreover, the HEL-AKO had higher cloning efficiency than HEL Control after ruxolitinib treatment. Conclusions Loss of function of ASXLI has an impact on cell biological function of HEL. Therefore, HEL- AKO cell line can be used to further explore the biological contribution of concomitant ASXLI in /4A2V617F mutated MPN.
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This study aimed to investigate the effects of Sargassum fusiforme polysaccharide (SFPS I, II, and III) on the apoptosis and regulation of human erythroleukemia (HEL) cells. The effect of different doses of SFPS on HEL cell growth was detected using the Cell Counting Kit-8 method, and apoptosis was detected by Hoechst staining. Cell cycle distribution and apoptosis were detected using flow cytometry. Expression of the cell cycle gene, p53, antiapoptotic genes, Bcl-xL and Bcl-2, and pro-apoptotic genes, Bax, Bad, and Caspase-3, as well as the expression of the corresponding proteins, were detected using real-time quantitative polymerase chain reaction (qPCR) and Western blot. The results showed that SFPS II and III decreased HEL cell viability and induced HEL cell apoptosis. Different concentrations of SFPS (I, II, and III) were detected that induced much less toxic effect in normal human embryonic lung (MRC-5) cells, and SFPS I increased cell proliferation, indicating its favorable selectivity towards cancer cells. The mechanism by which SFPS induced apoptosis was also found to be related to the induction of cell cycle arrest in the G/G phase and the increased expression of apoptosis-related genes and proteins. We concluded that SFPS induces HEL cell apoptosis, possibly via activation of the Caspase pathway, providing the theoretical basis for the development of SFPS-based anti-tumor drug products.
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Objective To observe the effect of Interferon-α2b on HEL cells (human erythroleukemia cell line) growth, apoptosis and JAK2 V617F mutation gene expression. Methods HEL cells were placed in RPMI1640 containing 10% FBS and incubated in a cell incubator. Cells in the logarithmic growth phasem were collected, adjusting the cell density to 1 × 105/mL for experimental research. The interferon concentration in five groups were 0, 5 × 105, 10 × 105, 50 × 105, 100 × 105 U/L, with different incubation time (0, 24, 72, 120 h), respectively. The cell growth status in different groups was observed in the inverted optical microscope; MTT was used to detect the inhibition of interferon on HEL cell proliferation. Cell apoptosis was detected by flow cytometry. Fluorescence quantitative PCR was used to detect the mutation gene of JAK2 V617F expression. Results Inhibition rates of Interferon on the HEL cell proliferation in 5 × 105 U/L, 10 × 105 U/L, 50 × 105 U/L, 100 × 105 U/L groups were 18.57%, 25.10%, 42.10%, 57.00%, respectively. JAK2 V617F/GAPDH by fluorescence quantitative was 1.556, 1.213, 0.870 respectively under the concentration of interferon 100 × 105 U/L for 24, 72, 120 h. Conclusions Interferon-α2b can inhibit HEL cells proliferation and induce HEL cells apoptosis. Increasing concentration of interferon increases HEL cell apoptosis rate. Interferon can inhibit JAK2 V617F expression of HEL cells in a dose-dependent manner.