[Optimization of determination methods of astragaloside â £ and analysis of contents of astragaloside â £ in different origin and grade].

A rapid and accurate method for determination of astragaloside Ⅳ was established,which was further applied to determine the contents of astragaloside Ⅳ in 87 batches of different origin and different grade of Astragali Radix. The ROC curve was used to analyze the contents of astragaloside Ⅳ in different origin. Simultaneous contents of astragaloside Ⅳ in different grade were compared with chemometrics. HPLC-ELSD method was used to determine the contents of astragaloside Ⅳ. A Vensil MP C18 column( 4. 6 mm×250 mm,5 µm) was used with acetonitrile-water( 32 ∶68) as the mobile phase at a flow rateof 1 m L·min-1. The column temperature was 25 ℃ with ELSD parameters as follows: gas flow rate was 2. 5 L·min-1,the drift tube heating temperature was set to 105 ℃,and the gain value was 4. 0. The optimized method avoided the problem that the consumable quality unstable and the recovery rate was not high. The contents determined by the optimized method were higher than the pharmacopoeia method,with less time and high recovery rate. The ROC curve analysis showed that there was no significant difference of contents of astragaloside Ⅳ between the top grade of Shanxi wild-simulated Astragali Radix top and the first grade of Gansu cultivated Astragali Radix. The contents of astragaloside Ⅳ in the second,third and fourth grade of Shanxi wild-simulated Astragali Radix was significantly higher than those of produced from Gansu.There was a significant negative correlation between the contents of astragaloside Ⅳ and grade in Shanxi Astragali Radix. While there was no correlation for Gansu Astragali Radix. This study provided the basis for the quality grade standard of Astragali Radix.

Cholesterol promotes the migration and invasion of renal carcinoma cells by regulating the KLF5/miR-27a/FBXW7 pathway.

Clear cell renal cell carcinoma (ccRCC) is the most common and lethal subtype of renal cell carcinoma. Accumulation of cholesterol and cholesterol ester is a remarkable feature of ccRCC. However, the effect of cholesterol on ccRCC remains unknown. Out results showed that cholesterol treatment significantly promoted cells migration and invasion in ccRCC. Mechanism analysis indicated that cholesterol induced KLF5 expression. KLF5 positively regulated the transcription of miR-27a, increasing miR-27a expression. MiR-27a directly targeted FBXW7 by binding to its 3'UTR, reducing FBXW7 expression. FBXW7 silencing further increased the expression of KLF5 and miR-27a, and promoted cells migration and invasion. These results suggested that cholesterol accelerated ccRCC cells migration and invasion by regulating KLF5/miR-27a/FBXW7 axis.

Plasmakinetic Vapor Enucleation of the Prostate with Button Electrode versus Plasmakinetic Resection of the Prostate for Benign Prostatic Enlargement >90 ml: Perioperative and 3-Month Follow-Up Results of a Prospective, Randomized Clinical Trial.

OBJECTIVE: To evaluate plasmakinetic vapor enucleation of the prostate (PVEP) with button electrode and plasmakinetic resection of the prostate (PKRP) in patients with urinary symptoms due to benign prostatic enlargement (BPE) >90 ml. METHODS: A total of 112 patients with symptomatic BPE were randomly assigned to either PKRP or PVEP prospectively from August 2012 to May 2014 in our department. Perioperative and postoperative data were investigated during a 3-month follow-up. RESULTS: PVEP was significantly superior to PKRP in terms of operation time (63.9 ± 7.7 vs. 78.1 ± 13.6 min, p < 0.001), hemoglobin loss (1.18 ± 0.30 vs. 1.63 ± 0.38 g/dl, p < 0.001), serum sodium decrease (2.9 ± 0.7 vs. 4.3 ± 0.8 mmol/l, p < 0.001), catheterization duration (49.3 ± 12.2 vs. 78.1 ± 14.8 h, p < 0.001) and hospital stay (100.2 ± 28.3 vs. 116.0 ± 29.2 h, p = 0.004). There were no statistical differences in blood transfusion between the two groups. In addition, there were no statistical differences in maximum urinary flow rate, International Prostate Symptom Score, postvoid residual urine volume, quality-of-life score, transient incontinence, and urethral stricture at 3 months postoperatively. CONCLUSIONS: PVEP with button electrode is an equally effective technique for treatment of large BPE with PKRP, with more safety and faster recovery. It may become the superior alternative to PKRP for patients with large BPE.

Inhibition of pancreatic lipase by coffee leaves-derived polyphenols: A mechanistic study.

The suppression of pancreatic lipase has been employed to mitigate obesity. This study explored the mechanism of coffee leaf extracts to inhibit pancreatic lipase. The ethyl acetate fraction derived from coffee leaves (EAC) exhibited the highest inhibitory capacity with a half-maximal inhibitory concentration (IC50) of 0.469 mg/mL and an inhibitor constant (Ki) of 0.185 mg/mL. This fraction was enriched with 3,5-dicaffeoylquinic acid (3,5-diCQA, 146.50 mg/g), epicatechin (87.51 mg/g), and isoquercetin (48.29 mg/g). EAC inhibited lipase in a reversible and competitive manner, and quenched its intrinsic fluorescence through a static mechanism. Molecular docking revealed that bioactive compounds in EAC bind to key amino acid residues (HIS-263, PHE-77, and SER-152) located within the active cavity of lipase. Catechin derivatives play a key role in the lipase inhibitory activity within EAC. Overall, our findings highlight the promising potential of coffee leaf extract as a functional ingredient for alleviating obesity through inhibition of lipase.

Characterization of kombucha prepared from black tea and coffee leaves: A comparative analysis of physiochemical properties, bioactive components, and bioactivities.

The utilization of coffee leaves in kombucha production has intrigued researchers; however, the lack of understanding regarding the characteristics of coffee leaf kombucha (CK) and its differentiation from black tea kombucha (BK) has impeded its application in the beverage industry. Therefore, this study aimed to characterize and compare the physiochemical properties, phytochemical compositions, antioxidant activity, and α-glucosidase inhibitory ability of kombucha prepared from the leaves of Coffea arabica (CK) and black tea (Camellia sinensis, BK) and their extracts (CT and BT). After fermentation, pH and the contents of total sugars, reducing sugars, and free amino acids of BK and CK were decreased, whereas the levels of total acids and organic acids, such as gluconic, lactic, and acetic acid were increased. Notably, the concentration of vitamin C in CK was 48.9% higher than that in BK. HPLC analysis exhibited that 5-caffeoylquinic acid in CT was significantly decreased by 48.0% in CK, whereas the levels of 3-caffeoylquinic acid and 4-caffeoylquinic acid were significantly increased after fermentation. The content of caffeine was significantly (p < 0.05) reduced by 9.5% and 22.0% in BK and CK, respectively, whereas the theobromine level was significantly increased in CK. Notably, CK has superior total phenolic and flavonoid contents and antioxidant activity than BK, whereas BK possesses higher α-glucosidase inhibitory capacity. Electronic nose analysis demonstrated that sulfur-containing organics were the main volatiles in both kombuchas, and fermentation significantly increased their levels. Our study indicates that coffee leaves are a promising resource for preparing kombucha. PRACTICAL APPLICATION: This article investigates the differences in physicochemical properties, bioactive constituents, antioxidant activity, and α-glucosidase inhibitory activity of kombucha preparation from black tea and coffee leaves. We have found that after fermentation BK had brighter soup color and higher α-glucosidase inhibitory capacity, whereas CK had higher levels of total phenols, flavonoids, vitamin C, and antioxidants and lower contents of sugars. This study provides valuable information for the preparation of CK with high-quality attributes and antioxidant activity.

Effective detection of Ag+, Hg2+ and dye adsorption properties studies of Ln-MOFs based on a benzimidazole carboxylic acid ligand.

Five new lanthanide metal-organic frameworks (Ln-MOFs), namely {[Ln(L)]·Cl}n, [Ln = Pr(1), Nd(2), Eu(3), Ho(4), Ce(5)], based on a benzimidazole carboxylic acid ligand [H2L = 2-(2-carboxyphenyl)-1H-benzo[d]imidazole-6-carboxylic acid] were synthesized by a solvothermal method. Ln-MOFs 1-5 have the same two-dimensional layered structures. Interestingly, 1-5 exhibit excellent adsorption performance to anionic dye Congo red (CR), with adsorption capacities of 2724 mg g-1, 2719 mg g-1, 2718 mg g-1, 327 mg g-1, and 2273 mg g-1, respectively. Adsorption kinetics experiments showed that this kind of adsorption belonged to chemisorption; the hydrogen bonding and π-π interactions between the 1-5 host and CR guest molecules resulted in a high adsorption capacity. Luminescence and sensing experiments showed that 5 can be considered a promising multifunctional fluorescent sensor with good reusability and a high sensitivity toward Ag+ and Hg2+ ions, with detection limits of 1.7 × 10-7 and 2.5 × 10-6 M, respectively.

Effect of acetabular morphological parameters applied in proximal femoral varus osteotomy on the treatment of developmental dysplasia of the hip in children.

BACKGROUND: The study was carried out based on the background that though proximal femoral varus osteotomy is one of the most effective methods in treating developmental dysplasia of the hip (DDH), its surgical efficiency and effect need to be improved. METHODS: The neck shaft angle and acetabular morphological parameters were measured before and after operation and during clinical follow-up. The Severin criteria and McKay criteria were both adopted to score the measurements. The distance from the center of the femoral head to the medial cutting bone (expressed as C value) and the shortened lengths of the proximal femur (expressed as Δ S) calculated by formula and obtained by actual measurement were compared. RESULTS: The neck shaft angle and acetabular morphological parameters after operation and during clinical follow-up were significantly lower than those before operation (P<0.05). The proportion of children with McKay scores of I, II, III, and IV after operation was 50.00%, 41.67%, 8.33%, and 0.00%, respectively; while the proportion of children with Severin scores of I, II, III, and IV was 55.00%, 38.33%, 6.67%, and 0.00%, respectively. There were no significant differences in the C value and Δ S obtained after formula calculation and actual measurement (P>0.05). CONCLUSIONS: The acetabular morphological parameters applied in proximal femoral varus osteotomy are of great value for the treatment of DDH in children.

Arsenic(V) removal behavior of schwertmannite synthesized by KMnO4 rapid oxidation with high adsorption capacity and Fe utilization.

Adsorption is a simple and efficient way for arsenic contamination purification in water, with a pressing challenge to find a cheap and efficient adsorbent. As a poorly crystalline Fe(III)-oxyhydroxy sulfate mineral, schwertmannite can be As(V) adsorbent because of its tunnel structure and low cost. However, the schwertmannite synthesized commonly by H2O2 rapid oxidation suffers from the low Fe utilization and limited As(V) adsorption capacity. In this research, the schwertmannite is synthesized by KMnO4. The results show that the Fe utilization can be improved from 40% to 56%, with the As(V) adsorption capacities double times better than those synthesized by H2O2 at pH 7 and 2. The As(V) adsorption mechanisms at different pHs and the reason for the improvement of As(V) adsorption capacity are thoroughly investigated. The FTIR and EDS images confirm that As(V) adsorption exchange with SO42- is the dominant mechanism at pH 7 and 2. At pH 11, the As(V) is mainly removed by surface complexation because the surface SO42- is exchanged by OH-. The intraparticle diffusion model fitting and XPS results further reveal that the tunnel structure built by Fe-SO4 in the KMnO4 oxidized schwertmannite is more stable, possibly resulting in the better As(V) adsorption performance.

SLC12A8 plays a key role in bladder cancer progression and EMT.

Bladder cancer is the most common malignant tumor of the urinary system. The intention of the present research is to explore the prognostic value and biological function of solute carrier family 12 member 8 (SLC12A8) in bladder cancer. The analysis based on the TCGA and ONCOMINE database revealed that the expression of SLC12A8 in bladder cancer was notably increased compared with the normal group. SLC12A8 expression was notably correlated with the age, pathological stage, T-stage, and lymph node metastasis of bladder cancer patients. Moreover, the patients' overall survival was notably shorter in the high SLC12A8 group. Compared with the control, SLC12A8 upregulation enhanced the proliferative, invasive, and migratory capacities of bladder cancer cells and promoted the expression of epithelial-mesenchymal transition (EMT) protein markers including ß-catenin, vimentin, snail, and slug, while reduced the expression of E-cadherin. In the case of downregulated SLC12A8 expression, the proliferative, invasive, and migratory capacities of bladder cancer cells and the expression of EMT protein markers presented the opposite trend. This study demonstrated that SLC12A8 was highly correlated with oncogenesis and progression of bladder cancer, indicating that SLC12A8 may be a meaningful biomarker for initial diagnosis and early treatment of bladder cancer.

Transmembrane protein ADAM29 facilitates cell proliferation, invasion and migration in clear cell renal cell carcinoma.

Abnormal expression of ADAM29 has been frequently reported in several cancers, however, its role in clear cell renal cell carcinoma (ccRCC) has not evaluated in detail. Herein, we attempt to determine the biological role and the action mechanism of ADAM29 in ccRCC. Bioinformatics analysis based on the ccRCC RNA-Seq dataset from TCGA database revealed that ADAM29 was up-expressed in ccRCC tissues by comparison with normal tissues. And a significant increase of ADAM29 expression was also observed in 3 ccRCC cell lines (UT33A, Caki-1, and786-O) in comparison with normal cell line. Besides, high level of ADAM29 was found to be connected with the poor prognosis and could be considered as an independent prognosticator for patients with ccRCC. Furthermore, functional experiments in vitro demonstrated that ADAM29 promoted the growth, invasion and migration of ccRCC cells. Moreover, Western blot assays indicated that ADAM29 was positively correlated with the level of proliferation-related proteins Cyclin D1 and PCNA and motion-related proteins MMP9 and Snail. Our data indicate that ADAM29 acts as an oncogene that increases tumour cells proliferation, invasion and migration partly by regulating the expression of Cyclin D1/PCNA/MMP9/Snail, suggesting that ADAM29 may become a prognosticator and therapeutic candidate for ccRCC.

Validation of the current prognostic models for nonmetastatic renal cell carcinoma after nephrectomy in Chinese population: a 15-year single center experience.

OBJECTIVES: To explore the applicability of the current prognostic models for nonmetastatic renal cell carcinoma in the Chinese population based on a single center experience. METHODS: Clinical and pathological variables of 653 nonmetastatic renal cell carcinoma patients were retrospectively reviewed. Seven models were used to predict the prognosis, including the Yaycioglu model, the Cindolo model, the University of California Los Angeles Integrated Staging System model, the stage, size, grade, and necrosis model, the Kattan nomogram, the Sorbellini nomogram and the Karakiewicz nomogram. Three different end-points were used for validation, including overall survival, cancer-specific survival, and recurrence-free survival. Survival was estimated using the Kaplan-Meier method. Discriminating ability was assessed using the Harrell's concordance-index. RESULTS: At the last follow up, 159 patients had died due to various causes, and disease recurrence occurred in 156 patients. The discriminating ability of all models was confirmed in the Chinese population. Nomograms discriminate better than algorithms, regardless of end-points. The Kattan nomogram was the most accurate, with the highest concordance-indexes of 0.752, 0.793 and 0.841 for overall survival, cancer-specific survival, and recurrence-free survival, respectively. CONCLUSIONS: The current prognostic models were developed and validated entirely based on Caucasian populations. This study defines the general applicability of the models for Chinese patients with nonmetastatic renal cell carcinoma treated with nephrectomy. The Kattan model was found to be the most accurate. The Cindolo model performed well in some situations, although only including clinical presentation and size of tumor. Therefore, models should be chosen according to different environments and purposes.

Comparison Between Thromboelastography and Conventional Coagulation Tests in Surgical Patients With Localized Prostate Cancer.

We aimed to examine hypercoagulable and hypocoagulable conditions in patients with prostate cancer using thromboelastography (TEG) and correlate TEG parameters with conventional coagulation test. The t test was used for comparing TEG parameters and routine coagulation results. Spearman rank-order correlation was used to describe the relationship of TEG and conventional tests. Sensitivity, specificity, positive predictive values, and negative predictive values were determined for bleeding and thrombosis. Totally, 20 patients had active bleeding postoperatively, 16 of whom showed hypocoagulation on TEG test and 9 of whom showed hypocoagulation by routine coagulation test ( P = .024). Overall, 60 patients did not have active bleeding postoperatively, 51 of whom showed hypercoagulation detected by TEG test and 42 of whom showed hypercoagulation found by routine coagulation test ( P = .040). Remarkably, patients had a little higher fibrinogen (FIB) compared to controls. There was no statistical difference in any of the conventional coagulation indexes between the groups. Correlation analysis showed that reaction time (R) and coagulation time (K) were positively correlated with the prothrombin time-international normalized ratio (PT-INR) and negatively correlated with FIB ( P < .001). Contrarily, α-angle and maximum amplitude (MA) were negatively correlated with PT-INR but positively correlated with FIB. Significantly, MA showed the strongest correlation with FIB and R exhibited the strongest correlation with PT-INR. Sensitivity and specificity for bleeding and thrombosis in TEG were higher than those in conventional coagulation test. Accordingly, TEG might be superior in evaluating hypercoagulation and detecting the risk of bleeding in patients with prostate cancer.

[An analysis for the phenotype and genotype of autosomal dominant polycystic kidney disease from two Chinese families].

OBJECTIVE: To scan for mutations of polycystic kidney disease 1 gene (PKD1) in Chinese population in order to find some features about Chinese patients and a better approach to detect mutations. METHODS: Twenty-five PKD-affected individuals from twenty-one unrelated genealogies and sixteen controls participated in the study. Thirty-five blood samples and six tissues were obtained after receiving informed consent and were in accordance with institutional ethical guidelines. Genomic DNA was isolated from peripheral blood using standard procedures. PCR amplification of genomic DNA was performed to generate the aimed fragments. Amplified fragments were analyzed by denaturing gradient gel electrophoresis (DGGE). A GC clamp was attached to the 5' primer. After that, the abnormal fragments were sequenced on freshly amplified specific PCR products with the dideoxynucleotide chain termination method. Sequencing was performed for all samples to evaluate DGGE. RESULTS: Aimed fragments of exons 44 and 45 were amplified. DGGE detected eleven abnormal PCR fragments. Two novel mutations were identified by sequencing, included one nonsense mutation (C12217T) and one frameshift (12431delCT). In addition, one polymorphism (A50747C) was identified. The mutation detection rate is 8% in our study. CONCLUSION: Two novel pathogenic mutations were detected, including one nonsense mutation (C12217T) and one frameshift (12431delCT).

Bone mesenchymal stem cells differentiate into myofibroblasts in the tumor microenvironment.

The aim of the present study was to investigate the tropism of mesenchymal stem cells (MSCs) to the tumor microenvironment, and to evaluate the feasibility of bone marrow mesenchymal stem cells differentiating into myofibroblasts in vitro. A total of 1 ml bone marrow was extracted from the greater trochanter of one male New Zealand rabbit, and MSCs were obtained by density gradient centrifugation and cultured routinely. The surface markers were analyzed by flow cytometry. A VX2 tumor was aseptically excised from another male New Zealand rabbit and primary cultured. The tropism of MSCs for 30% and 50% VX2 conditioned medium was determined by using Transwell migration assays. MSCs were incubated in 30% VX2 conditioned medium for 7 or 14 days. The messenger (m)RNA levels and protein expression of α-smooth muscle actin (α-SMA) and vimentin were measured by reverse transcription-polymerase chain reaction and western blotting. MSCs were observed to have a spindle shape. The cultured MSCs were cluster of differentiation (CD)44+, CD105+, CD106+ and CD34-. VX2 cells demonstrated a spindle or polygon shape. In the Transwell assay, it was observed that the migrated cells appeared more frequently in the 30% VX2 conditioned medium group compared with the other groups when microscopically examined, which was additionally confirmed by the results of a colorimetric assay. The mRNA levels and protein expression of α-SMA and vimentin significantly increased in the test group compared with the control group at 7 days (P<0.01), and further increased in the test group at 14 days (P<0.01). The results of the present study demonstrated that MSCs have tropism for the tumor microenvironment and furthermore, may differentiate into myofibroblasts in the tumor microenvironment in vitro. The present study suggested that MSCs may migrate to the tumor and subsequently differentiate into myofibroblasts due to the tumor microenvironment, which may lead to promotion of the growth of the tumor. The present study additionally suggested that MSCs may be the precursors of tumor/carcinoma-associated myofibroblasts.

The expression of Eg5 predicts a poor outcome for patients with renal cell carcinoma.

Eg5 is a member of the kinesin family of proteins, which associates with bipolar spindle formation in dividing tumor cells during mitosis. The aim of our study is to investigate the prognostic role of Eg5 expression in patients with renal cell carcinoma (RCC). RCC tissue specimens from 164 consecutively treated patients who underwent surgery between 2005 and 2011 were evaluated. The Eg5 expression was determined by immunohistochemistry, and correlated with clinicopathological parameters. The prognostic significance of Eg5 expression was explored using the univariate and multivariate survival analysis of 164 patients who were followed; one hundred and sixty-four tissue specimens "of patients" who were regularly followed with the mean 35.8 months (from 5 to 80 months). The expression of Eg5 was significantly associated with tumor nuclear grade (P = 0.019) and stage (P = 0.007), as well as tumor size (P = 0.033). In univariate analysis, Eg5 overexpression showed unfavorable influence on recurrence-free survival with statistical significance (P = 0.003). Clinical stage, nuclear grade and tumor size also showed strong statistical relation with adverse recurrence-free survival (P < 0.001). Multivariate analysis revealed that tumor stage, nuclear grade and Eg5 reactivity (P < 0.001, P = 0.002, P = 0.032) were identified as independent prognostic factors for recurrence-free survival in patients with RCC. In our opinion, the result of this study proved the relationship between Eg5 expression and worse clinical outcome in RCC. This finding suggested that Eg5 served as a prognostic factor, which could be useful to predict cancer evolution and provide appropriate treatments for RCC patients.

Tumor-infiltrating regulatory T cells are positively correlated with angiogenic status in renal cell carcinoma.

BACKGROUND: Immune cells within a tumor microenvironment have shown modulatory effects on tumor angiogenic activity. Renal cell carcinoma (RCC) is a hypervascular tumor that reportedly increases the frequency of regulatory T cells (Tregs) in tumor tissues. This study investigated the correlation between Tregs infiltration and angiogenic status in RCC. METHODS: Thirty-six patients with RCC were enrolled in the present study, and twenty age-matched healthy donors were included as the control. Tregs were defined as CD4(+)CD25(high)CD127(low/-) T cells. The frequency of Tregs in peripheral blood and tumor infiltrating lymphocytes (TILs) were determined by flow cytometry. The expression of vascular endothelial growth factor (VEGF) in surgical resection specimens were measured with a commercial enzyme-linked immunosorbent assay (ELISA) kit. Microvessel density (MVD) was calculated on slides stained with CD34 antibody. Spearman's rank correlation was performed to evaluate the correlation between the frequencies of Tregs in TILs and VEGF values, as well as between frequencies of Tregs and MVD determinations. RESULTS: Compared to healthy controls, the frequency of peripheral blood Tregs was significantly increased in patients with RCC (P < 0.05). The percentage of tumor-infiltrating Tregs was higher than that of peripheral blood Tregs in patients with RCC (P < 0.01). In addition, the frequency of tumor-infiltrating Tregs was shown to significantly correlate with the pathological stage (P < 0.05) and nuclear grade (P < 0.01). Importantly, a significant positive correlation was observed between the frequency of tumor-infiltrating Tregs and VEGF protein expression (r = 0.51, P < 0.05), as well as between frequencies of Tregs and MVD score (r = 0.39, P < 0.05). CONCLUSIONS: These observations suggest that the high pro-angiogenic status of RCC may be associated with the accumulation of Tregs in the local microenvironment. Angiogenesis networks may be connected with immune tolerance units and cooperate with each other to facilitate tumor growth and progression.