Anti-inflammatory material basis and mechanism of Artemisia stolonifera based on UPLC-Q-TOF-MS combined with network pharmacology and molecular docking / 中国中药杂志

This study aimed to explore the anti-inflammatory material basis and molecular mechanism of Artemisia stolonifera based on the analysis of the chemical components in different extracted fractions of A. stolonifera and their antioxidant and anti-inflammatory effects in combination with network pharmacology and molecular docking. Thirty-two chemical components were identified from A. stolonifera by ultra-performance liquid chromatography coupled to tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). Among them, there were 7, 21 and 22 compounds in water, n-butanol and ethyl acetate fractions, respectively. The antio-xidant capacity of different extracted fractions was evaluated by measuring their scavenging ability against 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl(DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonic acid)(ABTS) free radicals and total antioxidant capacity [ferric reducing antioxidant power(FRAP) assay]. The inflammatory model of RAW264.7 cells was induced by lipopolysaccharide(LPS), and the levels of nitrite oxide(NO), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) in the supernatant and the mRNA expression of related inflammatory factors in cells were used to evaluate the anti-inflammatory effects. The results revealed that ethyl acetate fraction of A. stolonifera was the optimal antioxidant and anti-inflammatory fraction. By network pharmacology, it was found that flavonoids such as rhamnazin, eupatilin, jaceosidin, luteolin and nepetin could act on key targets such as TNF, serine/threonine protein kinase 1(AKT1), tumor protein p53(TP53), caspase-3(CASP3) and epidermal growth factor receptor(EGFR), and regulate the phosphatidylinositol-3-kinase-protein kinase B(PI3K-AKT) and mitogen-activated protein kinase(MAPK) signaling pathways to exert the anti-inflammatory effects. Molecular docking further indicated excellent binding properties between the above core components and core targets. This study preliminarily clarified the anti-inflammatory material basis and mechanism of ethyl acetate fraction of A. stolonifera, providing a basis for the follow-up clinical application of A. stolonifera and drug development.

Analysis on Seed Quality Grading Standard of Belamcanda chinensis / 中国实验方剂学杂志

Objective:To study the quality inspection standard of seeds of <italic>Belamcanda chinensis</italic> and establish the quality grading standard of seeds of <italic>B. chinensis</italic>. Method:The purity of seeds of <italic>B. chinensis</italic> from different producing areas was analyzed，and the 1 000-grain weight，water content，viability and germination rate of seeds of <italic>B. chinensis</italic> with different diameters were determined after diameter classification. K-means was used for cluster analysis to preliminarily formulate the quality grading standard of seeds of <italic>B. chinensis</italic>. Result:There were obvious regional differences in the size of seeds of <italic>B. chinensis</italic>. The particle size of seeds of <italic>B. chinensis</italic> of Xiaochang，Dawu and Huanggang Academy of Agricultural Sciences in Hubei Province was larger and mostly between 4.5 mm to 5.5 mm. The seeds of<italic> B. chinensis </italic>from Tuanfeng，Qujing，Yunnan，and Anguo，Hebei province had smaller particle sizes than those from the other three producing areas，which were mostly between 3.5 mm to 5.0 mm. Arils in seeds of <italic>B. chinensis</italic> were removed by 10% NaClO to effectively break seed dormancy and significantly improve the seed germination rate. On this basis，the seed diameter was taken as the main grading index，and the seed moisture content，viability，germination rate and purity were taken as important reference indexes. The quality of seeds of <italic>B. chinensis</italic> was preliminarily divided into three grades，grade Ⅰ：seed diameter≥5.5 mm，moisture content≤10%，viability≥94%，germination rate≥60%，and cleanliness≥93%；grade Ⅱ：seed diameter≥4.5 mm，moisture content≤10%，viability≥90%，germination rate≥55%，and cleanliness≥85%；and grade Ⅲ：seed diameter≥3.5 mm，moisture content≤10%，viability≥84%，germination rate≥45%，cleanliness≥80%. Conclusion:In this study，the quality grading standard of seeds of <italic>B. chinensis</italic> was preliminarily established to provide reference for the quality evaluation of seeds of <italic>B. chinensis</italic> and the breeding of improved varieties. In addition，the maturity and the storage time of seeds of <italic>B. chinensis</italic> have a greater impact on the quality of seeds，so it is recommended to select fully mature（dark black） seeds and new seeds for production.

Active fractions of Camellia nitidissima inhibit non-small cell lung cancer via suppressing epidermal growth factor receptor / 中国中药杂志

The present study explored the effects and its underlying mechanisms of four active fractions of Camellia nitidissima(leaf polyphenols, leaf saponins, flower polyphenols, and flower saponins in C. nitidissima) in inhibiting the proliferation and migration of non-small cell lung cancer(NSCLC) by suppressing the epidermal growth factor receptor(EGFR). MTT assay was used to detect the effect of four active fractions on the proliferation of NCI-H1975 and HCC827 cells. Wound healing assay and Transwell assay were adopted to evaluate the effect of four active fractions on the migration of NSCLC. The effect of four active fractions on the enzyme activity of EGFR was detected. Molecular docking was carried out to explore the direct action capacity and action sites between representative components of the four active fractions and EGPR. Western blot assay was employed to investigate the effect of four active fractions on the protein expression in EGFR downstream signaling pathways. The results of the MTT assay indicated that the cell viability of NCI-H1975 and HCC827 cells was significantly inhibited by four active fractions at 50, 100, 150, and 200 μg·mL~(-1) in a dose-dependent manner. Wound healing assay and Transwell assay revealed that the migration of NCI-H1975 and HCC827 cells was significantly suppressed by four active fractions. In addition, the results of the protein activity assay showed that the enzyme activity of EGFR was significantly inhibited by four active fractions. The molecular docking results confirmed that various components in four active fractions possessed strong binding activity to EGFR enzymes. Western blot assay revealed that four active fractions down-regulated the protein expression of EGFR and its downstream signaling pathways. It is concluded that the four active fractions of C. nitidissima can inhibit NSCLC. The mechanism may be related to EGFR and its downstream signaling pathways. This study provides a new scientific basis for the clinical treatment of NSCLC with active fractions of C. nitidissima, which is of reference significance for further research on the anti-tumor mechanism of C. nitidissima.

Surgical efficacy after two different surgical methods on visual field and RNFL thickness in patients with glaucoma / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To compare the effects after two different surgical methods on visual field and retinal nerve fiber layer(RNFL)thickness in patients with glaucoma. <p>METHODS: Clinical data of 69 cases patients(82 eyes )with glaucoma between December 2015 to May 2017 in our hospital for anti-glaucoma surgery were collected. According to operation method, they were divided into EP group(EX-PRESS glaucoma shunt implantation)and XQ group(trabeculectomy). Intraocular pressure, anterior chamber depth(ACD), anterior chamber capacity(ACV), anterior chamber angle width(ACA), optical pupil diameter(PD), visual acuity, visual sensitivity index mean defect(MD), visual correction pattern standard deviation(CPSD), mean sensitivity(MS)and retinal nerve fiber layer thickness(RNFLT)were observed in the preoperative, postoperative 1mo and postoperative 3mo, and the postoperative complications were compared. <p>RESULTS: Hospitalization time of 3.08±0.42d in the EP group was significantly lower than 4.53±1.28d in the XQ group(<i>t</i>=6.124, <i>P</i><0.05). The visual recovery time of 3.26±0.30d in the EP group was significantly lower than 4.96±1.36d in the XQ group(<i>t</i>=6.920, <i>P</i><0.05). The ACA, ACD and ACV after surgery of two groups were significantly higher than those before operation, and the PD after surgery was lower than before operation, and the differences were not statistically significant between two groups after surgery(<i>P</i>>0.05).The postoperative RNFLT, MD and CPSD were all decreased in both groups(<i>P</i><0.05), and postoperative MS increased(<i>P</i><0.05); compared with XQ group, the postoperative CPSD after surgery in EP group was significantly reduced(<i>P</i><0.05).The total incidence of postoperative complications of EP group was 38%, which was significantly lower than 70% in XQ group, the difference was statistically significant(<i>χ</i>²=8.094, <i>P</i>=0.004). <p>CONCLUSION: Compared with trabeculectomy, EX-PRESS glaucoma shunt implantation is more effective to slow the progress of vision, and it is more conducive to recovery of the patient's vision after surgery with higher security. The improvement of the thickness of the retinal nerve fibers after EX-PRESS glaucoma shunt implantation is consistent with trabeculectomy.

Construction of recombinant adenovirus containing BMP-7 gene and its expression in proximal tubule epithelial cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus vector containing bone morphogenic protein-7 (BMP-7) gene and to identify its biological activities in proximal tubule epithelial cells (PTECs).</p><p><b>METHODS</b>Forty-six fragments of BMP-7 gene were obtained by PCR method and then were ligated to the full-length gene. The full length sequence of BMP-7 was subcloned into pBluescript II(+) vectors, and confirmed by sequencing. Double digested with Not I and Hind III, BMP-7 gene was inserted into pShuttle-CMV. EcoR I pre-linearized pShuttle plasmid was transformed into competence bacterium BJ5183 to obtain the recombinant adenovirus-BMP-7 by efficient homologous recombination. Then the AdBMP-7 was obtained by packaging Pac I linearized in 293 cells. Adenoviral titer was determined by adenovirus fluorescent detection kit. The protein expression of BMP-7 in PTECs was respectively detected by ELISA and Western blot. RT-PCR method was used for analyzing the alpha-smooth muscle action (alpha-SMA) expression in PTECs, which was treated consecutively with TGF-beta and AdBMP-7.</p><p><b>RESULTS</b>The recombinant plasmid AdBMP-7 was successfully generated, which increased BMP-7 protein expression levels in PTECs, and down-regulated TGF-beta-induced alpha-SMA expression.</p><p><b>CONCLUSION</b>The bioactive recombinant adenovirus AdBMP-7 has been successfully constructed, which may be effective in inhibition of chronic renal fibrosis.</p>

Primary chronic angle-closure glaucoma in younger patients / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To evaluate the clinical outcomes of management in younger patients with primary chronic angle-closure glaucoma (PCACG).METHODS: Thirty-eight patients (50 eyes) aged 40 or younger with confirmed diagnosis of PCACG in advanced or late stage who received surgical treatment in Zhongshan Ophthalmic Center from January 2000 to December 2005were retrospectively investigated. All patients underwent trabeculectomy. The mean follow-up was 23.6±7.5 months.Full ophthalmic examinations were performed. The clinical outcomes including clinical presentations, surgical results and complications were evaluated.RESULTS: The mean age of patients was 33.5±6.1 years old. There was a female preponderance (60.5%). The mean axial length was 22.4±3.5mm with 18.0% short axis of eyeball and 14% nanophthalmos. There was 60.0% fiat anterior chamber depth (＜1.9mm). Ultrasonic Biomicroscopy identified that plateau iris was the most common underlying etiology (80.6%). There was a statistically significant difference in intraocular pressure (IOP) reduction postoperativelyvs preoperatively (P＜0.001). Four eyes failed to control IOP and received second filtration surgery. The main postoperative complications included shallow anterior chamber (20.0%) and malignant glaucoma (12.0%).CONCLUSION: The younger PCACG patients in advanced or late stage can be effectively managed by trabeculectomy.They have more frequency of postoperative sustained shallow anterior chamber and malignant glaucoma. Careful ophthalmic examinations, delicate surgical procedures and well-managed technique of complications were suggested on younger PCACG patients.