The Herbal Formula Granule Prescription Mahuang Decoction Ameliorated Chronic Kidney Disease Which Was Associated with Restoration of Dysbiosis of Intestinal Microbiota in Rats.

Chronic kidney disease (CKD) has become a global health issue, and there is increasing evidence showing the beneficial roles of traditional Chinese medicine (TCM) in CKD treatment. Here, we studied the renoprotective role of Mahuang decoction, a famous TCM prescription, in a rat CKD model induced with the combination of doxorubicin and adenine. Our data showed that intragastric administration of Mahuang decoction inhibited the loss of bodyweight and attenuated proteinuria, serum creatinine, and blood urea nitrogen in CKD rats. Kidney histological analysis revealed decreased tubulointerstitial injury and fibrosis in CKD rats treated with Mahuang decoction accompanied with suppressed expression of TGF-ß1 and phosphorylated NF-κB/P65 (p-P65) as indicated by immunohistochemistry. ELISA analysis demonstrated reduced serum levels of proinflammatory cytokines TNFα and IL-6. Most importantly, intestinal microbiota analysis by 16s rRNA-seq showed that Mahuang decoction restored the impaired richness and diversity of intestinal microflora and recovered the disrupted microbial community through reducing the abundance of deleterious microbes and promoting the expansion of beneficial microbes in CKD rats. Collectively, our findings demonstrated that Mahuang decoction mitigated kidney functional and structural impairment in CKD rats which were associated with the restoration of dysbiosis of intestinal microbiota, implying its potential in clinical CKD treatment.

Circ-ZFR Promotes Progression of Bladder Cancer by Upregulating WNT5A Via Sponging miR-545 and miR-1270.

BACKGROUND: Bladder cancer is one of the most common cancers all over the world. CircZFR is a circular RNA and has been implicated in tumor generation and invasion. However, the exact role of circZFR in the development of bladder cancer (BCa) remains unknown. This study aimed to investigate the function of circZFR in BCa, and further to probe into the association between circ-ZFR, miR-545/miR-1270 and WNT5A. METHODS: The expression of circZFR in BCa was quantified by qRT-PCR and was positively correlated with the prognosis of BCa patients. Next, the stable knockdown of circZFR BCa cell lines was established and the resulting capacities of proliferation, migration and invasion were measured. The association of circZFR with miR-1270/miR-545 was predicted by circinteractome prediction, and was confirmed by luciferase assay as well as RNA pull down assay. Furthermore, miRNA inhibitors, WNT5A overexpression and Pearson correlation analysis were used to examine the relationship between circZFR, miR-1270/miR-545 and WNT5A. RESULTS: The expression of CircZFR was up-regulated both in BCa tissues and in BCa cell lines, and was positively correlated with patient survival rates. Blocking of circZFR's expression by RNA inhibitors suppressed the proliferation, migration and invasion of BCa cells both in vitro and in vivo. On the other hand, overexpression of target miRNA supported that circZFR directly interact with miR-545 and miR-1270. Moreover, we demonstrated that circZFR promotes the progression of BCa by upregulating WNT5A's expression via sponging miR-545 and miR-1270. CONCLUSIONS: CircZFR promotes the proliferation, migration and invasion of BCa cells by upregulating WNT5A signaling pathway via sponging miR-545 and miR-1270. These results provide new insights into the molecular mechanism of circZFR in BCa progression, and more important, a novel target for BCa clinical treatment.

Bone marrow mesenchymal stem cell derived exosomes miR-21-5p promotes proliferation, migration and invasion of prostate cancer PC-3 cell by downregulating PHLPP2 / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effects of exosome originated from bone marrow mesenchymal stem cell (BMSCs) on proliferation, migration and invasion of prostate cancer PC-3 cell and its mechanism. Methods: qPCR was used to detect the expression level of miR-21-5p in prostate cancer cell lines. The morphology of exosomes isolated from BMSCs was observed with an electron microscope. Western blotting was used to detect the expressions of exosome surface markers and the epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, N-cadherin and Vimentin). Dual luciferase reporter gene experiment was used to detect the targeted regulation relationship between miR-21-5p and PH domain leucine-rich repeat protein phosphatase 2 (PHLPP2). PC-3 cells were co-cultured with 10 μl BMSCs exosomes suspension (Exo group), transfected with sh-PHLPP2 or antagomiR, then CCK-8 and Transwell experiments were used to detect changesinproliferation,migrationandinvasionofPC-3cell.Results: miR-21-5p was highly expressed in prostate cancer PC-3 cell line. The exosomes in the supernatant of BMSCs culture fluid were successfully isolated, and the typical vesicle-like structures of exosomes were observed under transmission electron microscope. Exosomes expressed specific proteins such as CD9, CD63 and CD81. In the Exo group, the proliferation, invasion, migration, as well as the expressions of N-cadherin, Vimentin and miR-21-5p in PC-3 cells were significantly higher than those in the control group (all P<0.05). PHLPP2 is a target gene of miR-21-5p. Compared with the control group, the expression of PHLPP2 in PC-3 cells of Exo group and sh-PHLPP2 group was significantly reduced (0.66±0.09, 0.42±0.05 vs 1.09±0.08, all P<0.01); cell viability, invasion and migration were significantly improved (all P<0.01); and E-cadherin expression level was significantly reduced while N-cadherin and Vimentin expressions were significantly increased (both P<0.05). Conclusion: miR-21-5p is highly expressed in prostate cancer PC-3 cell line. BMSC exosome miR-21-5p can increase the proliferation, migration and invasion ability of PC-3 cells through targeted down-regulation of PHLPP2.

An optimized 5/6 nephrectomy mouse model based on unilateral kidney ligation and its application in renal fibrosis research.

5/6 Nephrectomy (PNx) on rat and mouse mimics renal failure after loss of kidney function in human, and it has been widely used in CKD researches. However, existing methods for PNx model construction present high mortality of animals after modeling due to hemorrhage and infection in or after surgery. Here, we report a novel and highly efficient PNx modeling method to simulate conventional 5/6 nephrectomy, which significantly reduced the mortality of animals and simplified the modeling procedures. In this novel modeling method, we directly ligated the upper and lower poles of left kidney after removal the right kidney 1 week later (l-PNx), which leads to necrosis of ligated upper and lower poles of the kidney and mimics the conventional 5/6 nephrectomy (c-PNx). After modeling 4 and 12 weeks, the serum creatinine, BUN and proteinuria levels were strongly increased in both c-PNx and l-PNx model. Importantly, compared with the c-PNx, l-PNx model present more severe renal fibrosis estimated by Masson staining, IHC and western blotting. The results showed that the protein levels of α-SMA were significantly increased in the kidney of c-PNx and l-PNx models, but more increase was found in l-PNx model. It is noteworthy that, compared with c-PNx model, the survival rate of l-PNx model was markedly increased. In summary, we established a novel and efficient 5/6 nephrectomy model, which can mimic conventional 5/6 nephrectomy to construct a renal fibrosis and renal failure mouse model, that is conducive to mechanism and treatment researches of CKD.

Deregulation of long noncoding RNA (TUG1) contributes to excessive podocytes apoptosis by activating endoplasmic reticulum stress in the development of diabetic nephropathy.

The objective of this study was to investigate the molecular mechanism of how TUG1 interferes with the expression of C/EBP homologous protein (CHOP), peroxisome-proliferator-activated receptor-γ coactivator-1 alpha (PGC-1α), which contributes to the development of diabetic nephropathy. Real-time polymerase chain reaction and western blot analysis were performed to explore the regulatory relationship among TUG1, CHOP, PGC-1α, and caspase-3. Terminal deoxynucleotidyl transferase dUTP nick-end labeling was performed to confirm TUG1 involved in diabetic nephropathy (DN) through influencing podocytes apoptosis. TUG1 was highly expressed in a cell following treatment with high glucose, and PGC-1α and cleaved caspase-3 levels were much lower, while CHOP level was much higher in high glucose group (HG), furthermore, CHOP inhibited PGC-1α expression. TUG1 negatively regulated CHOP expression, and positively regulated PGC-1α expression. Meanwhile, total caspase-3 level in cell treated with or without HG transfected with CHOP small interfering ribonucleic acid (siRNA), TUG1, and TUG1 siRNA showed no evident difference with their corresponding control, while CHOP siRNA and TUG1 evidently decreased, and TUG1 siRNA remarkably increased cleaved caspase-3 level in HG or normal glucose groups in comparison with corresponding control. TUG1 and PGC-1α levels were much lower, while CHOP level was much higher in participants diagnosed with DN. A higher level of CHOP protein and lower level of PGC-1α were observed in subjects diagnosed with DN. Finally, podocytes apoptosis in the DN group was significantly promoted compared with that in nondiabetic renal disease group. Our current study has suggested for the first time that the long noncoding RNA (lncRNA) TUG1 influenced podocytes apoptosis via mediating endoplasmic reticulum stress (ERS)-CHOP-PGC-1α signaling pathway in HG-induced DN.

Ethanol Extract of Root of Prunus persica Inhibited the Growth of Liver Cancer Cell HepG2 by Inducing Cell Cycle Arrest and Migration Suppression.

Liver cancer is the second most lethal cancer and hepatocellular carcinoma (HCC) is the primary cancer subgroup. However, the current chemotherapy agents remain ineffective and present wide side effects for advanced HCC patient. In this study, we investigated the antitumor role of ethanol extract of root of peach tree (Prunus persica (L.) Batsch and hereafter designated as TSG in short of its Chinese name), which is an important ingredient in Chinese medicine prescription, in liver cancer cell HepG2. By cell viability assay, we showed that addition of TSG in the culture medium inhibited the cell growth of HepG2 cells in a dose and time-dependent way. Cell cycle analysis indicated that TSG caused sustained M/G2 phase arrest. The expression of mitosis-related protein Cdc25c was impaired upon TSG treatment. Furthermore, wound healing assay demonstrated that TSG treatment notably suppressed the migration of HepG2 cells and the expression of extracellular matrix metalloprotease, MMP3 and MMP9. Most significantly, administration of TSG inhibited in vivo tumor growth in nude mice. Our findings suggested that TSG may serve as a source to isolate anti-HCC therapeutic ingredients.

Identification of diagnostic markers involved in the pathogenesis of gastric cancer through iTRAQ-based quantitative proteomics.

We provide detailed datasets from our analysis of proteins that are differentially expressed in gastric cancer tissues compared with adjacent normal gastric tissues, as identified by iTRAQ-based quantitative proteomics. Also included is a set of representative images of immunohistochemical staining of gastric cancer tissues showing four levels of expression of fatty acid binding protein (FABP1) and fatty acid synthase (FASN). The data presented in this paper support the research article "Quantitative proteomic analysis reveals that proteins required for fatty acid metabolism may serve as diagnostic markers for gastric cancer" (Jiang et al., 2017) [1]. We expect that the data will contribute to the identification of sensitive and specific biomarkers for early detection of gastric cancer.

Quantitative proteomic analysis reveals that proteins required for fatty acid metabolism may serve as diagnostic markers for gastric cancer.

BACKGROUND: Gastric cancer is one of the leading causes of cancer-related deaths worldwide. The sensitivities and specificities of current biomarkers for gastric cancer are insufficient for clinical detection, and new diagnostic tests are therefore urgently required. METHODS: A discovery set of gastric cancer and adjacent normal tissues were analyzed for differentially expressed proteins by labeling of peptide digests with isobaric tag for relative and absolute quantitation (iTRAQ) reagents followed by liquid chromatography-electrospray ionization-tandem mass spectrometry. A validation set of 70 pairs of gastric cancer and adjacent normal tissues were examined to confirm the expression levels of the potential biomarkers identified by iTRAQ labeling. RESULTS: We detected 431 proteins associated with 16 KEGG pathways that were differentially expressed in gastric cancer tissues, of which 224 were upregulated and 207 were downregulated in gastric cancer tissues. Coexpression of fatty acid binding protein (FABP1) and fatty acid synthase (FASN) in gastric cancer tissues (61.4% sensitivity and 77.1% specificity) was strongly associated with lymph node metastasis and Tumor, Node, Metastasis stage I/II. CONCLUSION: Quantitative proteomic analysis of gastric cancer tissues revealed that coexpression of FABP1 and FASN may serve as a biomarker for detection of early gastric cancer.

Bacterial IgA protease-mediated degradation of agIgA1 and agIgA1 immune complexes as a potential therapy for IgA Nephropathy.

Mesangial deposition of aberrantly glycosylated IgA1 (agIgA1) and its immune complexes is a key pathogenic mechanism of IgA nephropathy (IgAN). However, treatment of IgAN remains ineffective. We report here that bacteria-derived IgA proteases are capable of degrading these pathogenic agIgA1 and derived immune complexes in vitro and in vivo. By screening 14 different bacterial strains (6 species), we found that 4 bacterial IgA proteases from H. influenzae, N. gonorrhoeae and N. meningitidis exhibited high cleaving activities on serum agIgA1 and artificial galactose-depleted IgA1 in vitro and the deposited agIgA1-containing immune complexes in the mesangium of renal biopsy from IgAN patients and in a passive mouse model of IgAN in vitro. In the modified mouse model of passive IgAN with abundant in situ mesangial deposition of the agIgA-IgG immune complexes, a single intravenous delivery of IgA protease from H. influenzae was able to effectively degrade the deposited agIgA-IgG immune complexes within the glomerulus, demonstrating a therapeutic potential for IgAN. In conclusion, the bacteria-derived IgA proteases are biologically active enzymes capable of cleaving the circulating agIgA and the deposited agIgA-IgG immune complexes within the kidney of IgAN. Thus, the use of such IgA proteases may represent a novel therapy for IgAN.

Pharmacokinetics and tissue residues of hydrochloric acid albendazole sulfoxide and its metabolites in crucian carp (Carassius auratus) after oral administration.

The pharmacokinetics and residues elimination of hydrochloric acid albendazole sulfoxide (ABZSO) and its metabolites were studied in healthy crucian carp (Carassius auratus, 250 ± 30 g) kept at water temperatures of 10 °C and 25 °C. The concentrations of ABZSO and its metabolites concentration in plasma and tissues were determined using high-performance liquid chromatography (HPLC) using an ultraviolet detector. The results revealed that the plasma concentration of ABZSO in plasma was significantly higher than that of albendazole sulfone (ABZSO(2)), whereas albendazole-2-aminosulfone (ABZ-SO(2)NH(2)) was not detected. The plasma concentrations of ABZSO and its main metabolite ABZSO(2) concentration-time data were fitted using a single-compartment model at 10 °C and 25 °C. The absorption half-life (t1/2ka) of ABZSO was 3.86 h at 10 °C and 1.29 h at 25 °C, whereas the elimination half-life (t1/2ke) was 16.34 h at 10 °C and 6.72 h at 25 °C; the maximum plasma concentration (C(max)) and the time-point of maximum plasma concentration (T(p)) were calculated as 3.20 µg mL(-1) and 10.58 h at 10 °C, 4.39 µg mL(-1) and 3.80 h at 25 °C. The distribution volume (V(d)/F) of ABZSO was estimated to be 1.99 L kg(-1) at 10 °C and 1.53 L kg(-1) at 25 °C; the total body clearance (CL(b)) of ABZSO were computed as 0.08 and 0.19 L/(h kg) at 10 and 25 °C, respectively; the areas under the concentration-time curve (AUC) was 118.22 µg mL(-1)h at 10 °C and 63.12 µg mL(-1)h at 25 °C. The [Formula: see text] of ABZSO(2) was found to be 6.39 °C at 10 °C and 3.73 h at 25 °C, whereas the [Formula: see text] was 12.86 h at 10 °C and 6.56 h at 25 °C; the C(max) and T(p) of ABZSO(2) was calculated as 0.78 µg mL(-1) and 12.82 h at 10 °C, 1.03 µg mL(-1) and 7.04 h at 25 °C, respectively; the V(d)/F of ABZSO(2) were estimated to be 6.43 L kg(-1) at 10 °C and 4.61 Lkg(-1) at 25 °C; the CL(b) of ABZSO(2) were computed as 0.34 and 0.49 L/(h kg) at 10 °C and 25 °C, respectively; the AUC of ABZSO(2) were 28.86 µg mL(-1)h at 10 °C and 20.52 µg mL(-1)h at 25 °C. It was demonstrated that ABZSO(2) was the main metabolite of ABZSO. The concentrations of ABZSO and its main metabolite (ABZSO(2)) were detected in muscle, skin, liver and kidney, whereas ABZ-SO(2)NH(2) was only detected in liver and kidney. The ABZSO and it metabolite (ABZSO(2)) could still be detected at 4 d time-point after administration at both temperatures in all tissues. The results revealed that the depletion of ABZSO and its metabolite (ABZSO(2)) in crucian carp was slower with a long half-life time, especially at lower water temperature.