PCI-32765,an oral selective and irreversible inhibitor of Bruton tyrosine kinase (BTK),inhibits survival,activation,proliferation and migration of malignant B cells by blocking B cell receptor signaling pathway.PCI-32765 not only acts on malignant B cell,but also prevents resistance to chemical drugs.Therefore,PCI-32765 has broad prospects in the treatment of B cell malignancies.

Objective:To investigate the in vitro effect of arsenic trioxide (As2O3) alone and in combination with dexamethasone (DXM), etoposide (VP-16), methotrexate (MTX), bortezomib (BTZ), and suberoylanilide hydroxamic acid (SAHA) on the growth of human cutaneous T cell lymphoma (CTCL) cells Hut-78 and Hut-102. Methods:Hut-78 and Hut-102 cells were cultured with different concentrations of As2O3, DXM, VP-16, MTX, BTZ, and SAHA alone and As2O3 in combination with DXM, VP-16, MTX, BTZ, or SAHA for 48 h. The effects of the different samples on Hut-78 and Hut-102 cell proliferation were determined by MTT assay. Analyses using CalcuSyn software were performed to determine whether the combination of As2O3 with DXM, VP-16, MTX, BTZ, or SAHA in-duced synergistic cytoxicity. Results:As2O3, DXM, VP-16, MTX, BTZ, and SAHA alone significantly inhibited the growth of Hut-78 and Hut-102 cells in a dose-dependent manner, with a 50%inhibiting concentration of 5μmol/L, 500μg/mL, 2.5μg/mL, 1μg/mL, 10μmol/L, and 2.5μmol/L individually after 48 h of culture. As2O3 with DXM, VP-16, MTX, BTZ, or SAHA showed remarkable antitu-mor efficacy compared with that of individual applications. Conclusion:As2O3 alone or combined with DXM, VP-16, MTX, BTZ, or SAHA significantly inhibited Hut-78 and Hut-102 cell growth in vitro. This study demonstrated that As2O3 with DXM, VP-16, MTX, BTZ, or SAHA presents synergistic antitumor effects on CTCL cells and may be an optimal regimen in clinical trials of CTCL.

Objective:To detect the inhibitory effects of CAL-101, a selective inhibitor of phosphoinostitide-3'-kinase delta (PI3Kδ), on Burkitt's lymphoma cell line Raji and diffused large B-cell lymphoma cell line SUDHL-10 and elucidate its relative mechanism. Methods:Raji and SUDHL-10 cells were treated with various concentrations of CAL-101. Methyl thiazolyl tetrazolium (MTT) assay was performed to determine the inhibitory effect of CAL-101 on lymphoma cells, and cell apoptosis was measured by Annexin V/PI and DAPI staining. Migration assays were performed with transwell to detect the migration of lymphoma cells derived from the stromal cell line HK. Western blot was used to detect the phosphorylation status of the ERK pathway. MTT and CalcuSyn software analyses were preformed to detect whether or not combining CAL-101 with bortezomib induces synergistic cytoxicity. Results:CAL-101 at con-centrations of 5, 10, 15, and 20μmol/L inhibited cell proliferation in a dose-dependent manner. The proliferation rates of the Raji cells treated with 5, 10, 15, and 20μmol/L for 48 h were 29.17%± 1.23%, 38.15%± 1.51%, 46.46%± 1.78%, and 55.8%± 2.01%, respec-tively, which were significantly higher (P<0.05) than that of the control group (1.15% ± 0.02%). Similar results were found in the SUDHL-10 cells after treatment with CAL-101 (P<0.05). CAL-101 also exerted an apoptotic effect on the lymphoma cells. The apop-totic rates of the Raji cells treated with CAL-101 for 21 h were 22.69%± 3.83%and 49.96%± 7.36%, respectively, which were signifi-cantly higher (P<0.05) than that of the control group (5.23%± 2.04%). Similar results were found in the SUDHL-10 cells (P<0.05). Treatment with 5 and 10 μmol/L CAL-101 dose-dependently inhibited the migration activity of lymphoma cells to stromal cells (P<0.05). Western blot analysis showed that the expression level of ERK phosphorylation protein was significantly downregulated in the cells treated with CAL-101. A synergistic effect between CAL-101 and bortezomib was verified. That is, these two drugs can signifi-cantly inhibit the proliferation of lymphoma cells with CI values less than 1. Conclusion:The PI3Kδ-specific inhibitor CAL-101 sup-pressed the proliferation of Raji and SUDHL-10 cells, induced apoptosis, and inhibited stromal cell-derived migration. This inhibitory effect may be induced by blocking the ERK pathway. Overall, our study indicated that CAL-101 is a novel and potential agent in the therapeutic strategy against aggressive B-cell lymphoma.

Objective:To observe the effects of acupuncture and moxibustion at Feishu(BL13)on inflammatory responses and intestinal short-chain fatty acids(SCFAs)in rats with asthma. Methods:Fifty-six Sprague-Dawley rats were randomly divided into a normal group(16 rats)and a modeling group(40 rats).Rats in the modeling group were subjected to establishing asthma models using ovalbumin(OVA).Model evaluation was conducted using 4 rats from each group.The remaining rats that successfully developed asthma were then randomly divided into a model group,an acupuncture group,and a moxibustion group,with 12 rats in each group.Rats in the acupuncture group received acupuncture treatments,and those in the moxibustion group received moxibustion treatments,both at Feishu(BL13)for 30 min.Following the treatments,the rats were exposed to atomization excitation with a 1%OVA solution for 20 min daily for 14 consecutive days.At the end of the experiment,inflammatory markers in the rats'peripheral blood were analyzed using a biochemical method.In addition,inflammatory cells in the bronchoalveolar lavage fluid(BALF)were counted using Wright-Giemsa staining.The lung tissue of rats was examined under a light microscope after staining with hematoxylin-eosin to observe morphological or pathological changes.Furthermore,real-time fluorescence quantitative polymerase chain reaction was utilized to measure the mRNA expression of inflammatory factors in the lung tissue.Lastly,the concentration of SCFAs in the rat's feces was determined using gas chromatography-hydrogen flame ionization. Results:The levels of eosinophils(Eos),neutrophils(Neu),and lymphocytes(Lym)in the peripheral blood,as well as Eos and Neu in the BALF,and the expression of interleukin(IL)-4,IL-5,IL-13,IL-33,and thymic stromal lymphopoietin(TSLP)mRNAs in the lung tissue were all found to be significantly increased(P<0.05 or P<0.01);the lung tissue structure displayed severe injuries;the levels of acetic acid,propionic acid,isobutyric acid,butyric acid,and valeric acid in the feces decreased significantly in the model group(P<0.05 or P<0.01).Compared with the model group,the peripheral blood levels of Eos,Neu,and Lym,as well as Eos in the BALF,and the mRNA expression levels of IL-4 and IL-5 in the lung tissue decreased significantly in both the acupuncture group and the moxibustion group(P<0.05 or P<0.01).This reduction was accompanied by alleviated pathological damage in the lung tissue.Additionally,there were significant increases in the levels of acetic acid,propionic acid,isobutyric acid,and butyric acid in the feces in both the acupuncture group and the moxibustion group(P<0.05 or P<0.01).In the acupuncture group,the expression levels of Lym in the BALF and IL-13 mRNA in the lung tissue decreased significantly(P<0.05 or P<0.01).In the moxibustion group,the mRNA expression levels of IL-33 and TSLP in the lung tissue also reduced significantly(P<0.05 or P<0.01).Furthermore,the level of valeric acid in the feces increased notably in the moxibustion group(P<0.01).Compared with the acupuncture group,it was found that the mRNA levels of IL-5 and IL-13 in the lung tissue,as well as the acetic acid level in the feces,were significantly higher in the moxibustion group(P<0.05 or P<0.01). Conclusion:Both acupuncture and moxibustion were effective in reducing abnormal inflammation and regulating intestinal SCFAs in asthma model rats.Acupuncture demonstrated superiority in inhibiting pro-inflammatory factors,particularly IL-5 and IL-13,while moxibustion exhibited better regulation on intestinal metabolites SCFAs,especially acetic acid.

Objective To investigate the effect of CAL?101, a selective inhibitor of PI3Kδ, in combination with bortezomib on the proliferation and apoptosis in human mantle cell lymphoma cell lines Z138, HBL?2 and Jeko?1 in vitro, to explore its mechanisms and provide the foundation for effective treatment strategies against mantle cell lymphoma. Methods MTT assay was applied to detect the inhibitory effects of CAL?101 and bortezomib either alone or combined on Z138, HBL?2 and Jeko?1 cells. Calcusyn software was used to analyze the synergistic cytotoxicity. Western blot was used to detect the expression of PI3K?p110σ and p?Akt, Akt, p?ERK and ERK proteins after the cells were exposed to different concentrations of CAL?101. Flow cytometry was employed to assess the apoptosis rate. NF?κB kit was used to determine the changes of location of NF?κB P65, and Western blot was applied to detect the level of caswpase?3 and the phosphorylation of Akt in different groups. Results CAL?101 and BTZ inhibited the proliferation of Z138, HBL?2 and Jeko?1 cells in a dose? and time?dependent manner. CAL?101/BTZ combination induced significantly synergistic cytotoxicity in the MCL cells. The results of Western blot assay showed that CAL?101 significantly blocked the phosphorylation of Akt and ERK in the MCL cell lines. In addition, CAL?101 combined with BTZ induced pronounced apoptosis (P<0.01). For example, after the Z138 cells exposed to the drugs for 48 h, the apoptosis rates of the control, CAL?101, BTZ and CAL?101+BTZ groups were:(2.6±1.8)%, (40.0±3.0)%, (34.0±1.0)%, and (67.4±1.0)%, respectively; and when drug treatment was given to HBL?2 cells over 96 h, the apoptosis rates of these four cell groups were (7.4±0.6)%,(30.7±5.7)%, (12.0±1.0)%, and (85.0±4.0)%, respectively. The combination therapy contributed to the enhanced inactivity of nuclear factor?κB ( NF?κB) and Akt inactivation in the MCL cell lines (P<0.05), however, the casepase?3 activity was up?regulated. Conclusions The combination of CAL?101 and bortezomib is muchmore effective in inhibiting proliferation and promoting apoptosis of mantle cell lymphoma cell lines ( Z138, HBL?2 and Jeko?1) , which may be mediated through inhibiting PI3K/Akt signaling pathway and the transcription of NF?κB.

Objective To investigate the effect of CAL?101, a selective inhibitor of PI3Kδ, in combination with bortezomib on the proliferation and apoptosis in human mantle cell lymphoma cell lines Z138, HBL?2 and Jeko?1 in vitro, to explore its mechanisms and provide the foundation for effective treatment strategies against mantle cell lymphoma. Methods MTT assay was applied to detect the inhibitory effects of CAL?101 and bortezomib either alone or combined on Z138, HBL?2 and Jeko?1 cells. Calcusyn software was used to analyze the synergistic cytotoxicity. Western blot was used to detect the expression of PI3K?p110σ and p?Akt, Akt, p?ERK and ERK proteins after the cells were exposed to different concentrations of CAL?101. Flow cytometry was employed to assess the apoptosis rate. NF?κB kit was used to determine the changes of location of NF?κB P65, and Western blot was applied to detect the level of caswpase?3 and the phosphorylation of Akt in different groups. Results CAL?101 and BTZ inhibited the proliferation of Z138, HBL?2 and Jeko?1 cells in a dose? and time?dependent manner. CAL?101/BTZ combination induced significantly synergistic cytotoxicity in the MCL cells. The results of Western blot assay showed that CAL?101 significantly blocked the phosphorylation of Akt and ERK in the MCL cell lines. In addition, CAL?101 combined with BTZ induced pronounced apoptosis (P<0.01). For example, after the Z138 cells exposed to the drugs for 48 h, the apoptosis rates of the control, CAL?101, BTZ and CAL?101+BTZ groups were:(2.6±1.8)%, (40.0±3.0)%, (34.0±1.0)%, and (67.4±1.0)%, respectively; and when drug treatment was given to HBL?2 cells over 96 h, the apoptosis rates of these four cell groups were (7.4±0.6)%,(30.7±5.7)%, (12.0±1.0)%, and (85.0±4.0)%, respectively. The combination therapy contributed to the enhanced inactivity of nuclear factor?κB ( NF?κB) and Akt inactivation in the MCL cell lines (P<0.05), however, the casepase?3 activity was up?regulated. Conclusions The combination of CAL?101 and bortezomib is muchmore effective in inhibiting proliferation and promoting apoptosis of mantle cell lymphoma cell lines ( Z138, HBL?2 and Jeko?1) , which may be mediated through inhibiting PI3K/Akt signaling pathway and the transcription of NF?κB.

Diabetic nephropathy (DN) is a clinical syndrome characterized by persistent proteinuria and progressive decline in renal function, and is one of the microvascular complications of diabetes. With the in-depth understanding of the pathogenesis of DN, the role of intestinal flora imbalance in the disease has been found clinically. This suggests that restoring the host′s healthy gut flora may be a means of improving DN. In fact, recent studies have shown that many of the drugs currently used to treat DN affect gut microbiota composition. In this review, intestinal flora is regarded as one of the main factors affecting the development of DN, and DN therapy targeting intestinal flora is summarized to provide new ideas for the diagnosis and treatment of DN.

ObjectiveTo explore the possible mechanism of moxibustion at "Feishu （BL13）" and "Tianshu （ST25）" in treatment of bronchial asthma. MethodsA total of 48 SD rats were randomly divided into normal group （n=12） and modeling group （n=36）. The bronchial asthma rat model was established by sensitization with ovalbumin （OVA） injection and aerosol provocation. Thirty-two successfully modeled rats were further randomly divided into four groups including model group， Feishu group， Tianshu group， and Feishu-Tianshu group， with 8 rats in each group. Rats in the normal group and the model group were tied and fixed without intervention， while those in the Feishu group received moxibustion at bilateral of "Feishu （BL13）" for 30 minutes； rats in the Tianshu group received moxibustion at bilateral "Tianshu （ST25）" for 30 minutes， and those in the Feishu-Tianshu group received moxibustion at both "Feishu （BL13）" and "Tianshu （ST25）" bilaterally for 15 minutes each. One hour after the intervention， 1% OVA solution was aerosolized for 20 minutes in all groups except the normal group， which was given the same volume of 0.9% sodium chloride solution instead of OVA solution for aerosol stimulation. The above interventions were performed once daily for 14 days. Behavioral observations were performed after modeling and during the interventions. The samples were collected 24h after the last intervention. HE and Masson staining were used to observe pathological morphological changes of lung tissues， and the percentage of collagen fiber deposition area was counted. The levels of leukocyte differentiation antigen11b （CD11b）， leukocyte differentiation antigen 40 （CD40）， leukocyte differentiation antigen 86 （CD86）， and programmed death ligand 2 （PD-L2） in serum， as well as the expression of interleukin-8 （IL-8）， interleukin-11 （IL-11）， interleukin-27 （IL-27） in lung tissue， were detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the expression of matrix metalloproteinase-9 （MMP-9）， matrix metalloproteinase inhibitor 1 （TIMP-1） and transforming growth factor β （TGF-β） proteins in lung tissue. The content of six short-chain fatty acids （SCFAs） including acetic acid， propionic acid， isobutyric acid， butyric acid， valeric acid， and hexanoic acid in feces was detected by gas chromatography-mass spectrometry. ResultsCompared to the normal group， the rats in the model group gradually showed mental depression or restlessness， dull hair， slow activity， reduced food intake， unformed stool， accompanied by symptoms of shortness of breath and wheezing. The pathological results showed severe abnormalities in lung tissue structure in the model group， including extensive infiltration of inflammatory cells around the bronchi， thickening of the airway smooth muscle layer， and substantial deposition of collagen fibers. Significant increases were observed in the levels of serum CD11b， CD40， CD86， and PD-L2， levels of IL-8， IL-11， and IL-27 in the lung tissue， as well as protein expression levels of MMP-9， TGF-β， and TIMP-1 in lung tissue， while the fecal levels of acetic acid， propionic acid， isobutyric acid， butyric acid， valeric acid， and n-caproic acid significantly decreased （P＜0.05 or P＜0.01）. Compared to those in the model group， the spirit， hair， activity， drinking and eating condition， shortness of breath， and wheezing symptoms of rats in the Feishu group， Tianshu group， and Feishu-Tianshu group were improved； the stool was basically formed， and the pathological morphology of lung tissue were improved； the levels of serum CD11b， CD40， CD86 and PD-L2， the levels of IL-8 and IL-27 in the lung tissue， the percentage of collagen fiber deposition area， and the TGF-β protein expression notably decreased； content of IL-11 and MMP-9 in the lung tissue and protein expression of T1MP-1 in Feishu group and Feishu-tianshu group significantly decreased； content of six SCFAs in the Feishu-Tianshu group increased （P＜0.05 or P＜0.01）. Compared to those in the Feishu group， the percentage of collagen fiber deposition area and TIMP-1 protein expression in lung tissue in the Feishu-Tianshu group significantly decreased， while the fecal levels of acetic acid and butyric acid notably increased （P＜0.05 or P＜0.01）. Compared to those in the Tianshu group， the serum level of CD40 in the Feishu-Tianshu group was significantly reduced， and the percentage of collagen fiber deposition area， the content of IL-11， and the protein expressions of MMP-9， TGF-β and TIMP-1 in the lung tissue notably decreased， while the fecal levels of acetic acid， propionic acid， and butyric acid significantly increased （P＜0.05 or P＜0.01）. ConclusionMoxibustion at "Feishu （BL13）" and "Tianshu （ST25）" exhibits a favorable therapeutic effect on airway remodeling in bronchial asthma rats， and the combined application of "Feishu （BL13）" and "Tianshu （ST25）" acupoints demonstrates a synergistic effect. The mechanism may be related to the regulation of intestinal SCFAs content， influencing the differentiation of immune cells， and reducing airway inflammation.

OBJECTIVETo investigate the effect of CAL-101, a selective inhibitor of PI3Kδ, in combination with bortezomib on the proliferation and apoptosis in human mantle cell lymphoma cell lines Z138, HBL-2 and Jeko-1 in vitro, to explore its mechanisms and provide the foundation for effective treatment strategies against mantle cell lymphoma.

METHODSMTT assay was applied to detect the inhibitory effects of CAL-101 and bortezomib either alone or combined on Z138, HBL-2 and Jeko-1 cells. Calcusyn software was used to analyze the synergistic cytotoxicity. Western blot was used to detect the expression of PI3K-p110σ and p-Akt, Akt, p-ERK and ERK proteins after the cells were exposed to different concentrations of CAL-101. Flow cytometry was employed to assess the apoptosis rate. NF-κB kit was used to determine the changes of location of NF-κB P65, and Western blot was applied to detect the level of caswpase-3 and the phosphorylation of Akt in different groups.

RESULTSCAL-101 and BTZ inhibited the proliferation of Z138, HBL-2 and Jeko-1 cells in a dose- and time-dependent manner. CAL-101/BTZ combination induced significantly synergistic cytotoxicity in the MCL cells. The results of Western blot assay showed that CAL-101 significantly blocked the phosphorylation of Akt and ERK in the MCL cell lines. In addition, CAL-101 combined with BTZ induced pronounced apoptosis (P < 0.01). For example, after the Z138 cells exposed to the drugs for 48 h, the apoptosis rates of the control, CAL-101, BTZ and CAL-101 + BTZ groups were: (2.6 ± 1.8)%, (40.0 ± 3.0)%, (34.0 ± 1.0)%, and (67.4 ± 1.0)%, respectively; and when drug treatment was given to HBL-2 cells over 96 h, the apoptosis rates of these four cell groups were (7.4 ± 0.6)%, (30.7 ± 5.7)%, (12.0 ± 1.0)%, and (85.0 ± 4.0)%, respectively. The combination therapy contributed to the enhanced inactivity of nuclear factor-κB (NF-κB) and Akt inactivation in the MCL cell lines (P < 0.05), however, the casepase-3 activity was up-regulated.

CONCLUSIONSThe combination of CAL-101 and bortezomib is muchmore effective in inhibiting proliferation and promoting apoptosis of mantle cell lymphoma cell lines (Z138, HBL-2 and Jeko-1), which may be mediated through inhibiting PI3K/Akt signaling pathway and the transcription of NF-κB.

Antineoplastic Agents ; pharmacology ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Boronic Acids ; Bortezomib ; pharmacology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Class Ia Phosphatidylinositol 3-Kinase ; antagonists & inhibitors ; Dose-Response Relationship, Drug ; Drug Synergism ; Formazans ; Humans ; Lymphoma, Mantle-Cell ; drug therapy ; pathology ; MAP Kinase Signaling System ; drug effects ; NF-kappa B ; metabolism ; Neoplasm Proteins ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; drug effects ; Proto-Oncogene Proteins c-akt ; metabolism ; Purines ; administration & dosage ; pharmacology ; Pyrazines ; Quinazolinones ; administration & dosage ; pharmacology ; Signal Transduction ; Software ; Tetrazolium Salts

OBJECTIVETo elucidate the impact of Hes1 on the proliferation and apoptosis of acute myeloid leukemia (AML) cells.

METHODSThe expression levels of Hes1 and p21 in AML patient samples and myeloid leukemia cell lines were analyzed by real-time PCR. Hes1 was up-regulated by retrovirus transfection in AML cell lines and the proliferation capacity were assayed by MTT, cell cycle by Hoechst/PY, apoptosis by AnnexinV.

RESULTSThe expression of Hes1 in primary AML cells and HL-60, U937, KG1a cell lines were 0.67 ± 0.24, 0.59 ± 0.43, 0.42 ± 0.03, and 0.32 ± 0.26, respectively, and p21 were 0.54 ± 0.01, 0.44 ± 0.12, 0.36 ± 0.12, and 0.59 ± 0.43, respectively. Hes1 expression levels after transduction in HL-60, U937, KG1a were 4.9 ± 0.2, 5.2 ± 0.4, 5.8 ± 0.5, respectively. Induced activation of Hes1 led to AML cells growth arrest and apoptosis, which was associated with an enhanced p21 expression. Besides, activated Hes1 led to AML cells growth inhibition in vivo.

CONCLUSIONHes1 could mediate growth arrest and apoptosis in AML cells, which may be a novel target for AML.

Apoptosis ; Basic Helix-Loop-Helix Transcription Factors ; Cell Cycle ; Cell Line, Tumor ; Homeodomain Proteins ; Humans ; Leukemia, Myeloid, Acute ; Transcription Factor HES-1 ; Up-Regulation