Salidroside attenuates hypoxia-induced pulmonary arterial smooth muscle cell proliferation and apoptosis resistance by upregulating autophagy through the AMPK-mTOR-ULK1 pathway.

BACKGROUND: Recent studies have shown that both adenosine monophosphate activated protein kinase (AMPK) and the mammalian target of rapamycin (mTOR) are energy sensors and are related to autophagy. Our recent reports have shown that salidroside can exert protective effects against hypoxia-induced pulmonary arterial smooth muscle cell (PASMC) proliferation and apoptosis resistance through the AMPK pathway. This study aims to explore the relationship among AMPK, mTOR and ULK1 in PASMCs under hypoxic conditions and to investigate whether the protective effects of salidroside are related to the autophagic cell death pathway. METHODS: Rat PASMCs were cultured and divided into five groups: the normoxia, hypoxia, hypoxia + MHY1485 (mTOR agonist), hypoxia + rapamycin (mTOR inhibitor) and hypoxia + salidroside groups. Hypoxic cells were treated as indicated for 24 h. Cell viability was evaluated by the CCK-8 assay. Cell apoptosis was measured by the TUNEL assay. The autophagy flux of PASMCs was evaluated with tandem mRFP-GFP fluorescence microscopy. Autophagosomes were detected by electron microscopy. Protein expression of LC3, p62, AMPK, P-AMPK (Thr 172), P-ULK1 (Ser 555 and Ser 317), mTOR, P-mTOR (Ser 2448), ULK1 and P-ULK1 (Ser 757) was detected by western blot assay. RESULTS: PASMC proliferation and apoptosis resistance were observed under hypoxic conditions. Autophagy flux, the number of autophagosomes and the LC3II/LC3I ratio were increased in the hypoxia group compared with the normoxia group, whereas p62 expression was decreased. Treatment with rapamycin or salidroside reversed hypoxia-induced PASMC proliferation and apoptosis resistance and further increased autophagy flux, autophagosome levels and the LC3II/LC3I ratio but decreased p62 expression. Treatment with MHY1485 reversed hypoxia-induced PASMC apoptosis resistance and decreased autophagy flux as well as increased autophagosome levels, the LC3II/LC3I ratio and p62 expression. P-AMPK (Thr 172) and P-ULK1 (Ser 555) of the AMPK-ULK1 pathway were increased in the hypoxia group and were further increased in the salidroside group. Rapamycin and MHY1485 had no effect on either P-AMPK (Thr 172) or P-ULK1 (Ser 555). Phosphorylation of ULK1 at serine 317 did not significantly affect the five groups. Furthermore, P-mTOR (Ser 2448) and P-ULK1 (Ser 757) of the AMPK-mTOR-ULK1 pathway were decreased in the hypoxia group and were further decreased in the salidroside group. MHY1485 increased the expression of both P-mTOR(Ser 2448) and P-ULK1(Ser 757), whereas rapamycin had the opposite effect. CONCLUSIONS: Salidroside might inhibit hypoxia-induced PASMC proliferation and reverse apoptosis resistance via the upregulation of autophagy through both the AMPKα1-ULK1 and AMPKα1-mTOR-ULK1 pathways.

Determination of arsenic species in Solanum Lyratum Thunb using capillary electrophoresis with inductively coupled plasma mass spectrometry.

A simple and highly efficient interface to couple capillary electrophoresis with inductively coupled plasma mass spectrometry by a microflow polyfluoroalkoxy nebulizer and a quadruple ion deflector was developed in this study. By using this interface, six arsenic species, including arsenite, arsenate, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, and arsenocholine, were baseline-separated and determined in a single run within 11 min under the optimized separation conditions. The instrumental detection limit was in the range of 0.02-0.06 ng/mL for the six arsenic compounds. Repeatability expressed as the relative standard deviation (n = 5) of both migration time and peak area were better than 2.5 and 4.3% for six arsenic compounds. The proposed method, combined with a closed-vessel microwave-assisted extraction procedure, was successfully applied for the determination of arsenic species in the Solanum Lyratum Thunb samples from Anhui province in China with the relative standard deviations (n = 5) ≤4%, method detection limits of 0.2-0.6 ng As/g and a recovery of 98-104%. The experimental results showed that arsenobetaine was the main speciation of arsenic in the Solanum Lyratum Thunb samples from different provinces in China, with a concentration of 0.42-1.30 µg/g.

Baicalin attenuates bleomycin-induced pulmonary fibrosis via adenosine A2a receptor related TGF-ß1-induced ERK1/2 signaling pathway.

BACKGROUND: Baicalin has been reported to have anti-fibrosis effect; however, its mechanism still remains to be elucidated. Adenosine A2a receptor (A2aR) is a novel inflammation regulator, and transforming growth factor-ß1 (TGF-ß1)-induced extracellular signal regulated kinase1/2 (ERK1/2) signaling pathway plays an important role in idiopathic pulmonary fibrosis (IPF). This study was to explore the relationship of A2aR and TGF-ß1-induced ERK1/2 in bleomycin (BLM)-induced pulmonary fibrosis in mice, and to investigate whether A2aR mediate the anti-fibrosis effect of Baicalin on BLM-induced pulmonary fibrosis. METHODS: The A2aR-/- and A2aR+/+ mice were respectively divided into three groups: control group, model group, baicalin group. Pulmonary fibrosis was induced in mice of model groups by intratracheal instillation of bleomycin, and baicalin was administered in mice of baicalin groups daily for 28 days. Histopathological and ultrastructural changes of lung tissues were evaluated. Lung coefficient and the levels of hydroxyproline (HYP) in lung tissues were measured at the same time. The levels of serum TGF-ß1 were measured by ELISA. The expression of TGF-ß1, ERK1/2, p-ERK1/2 and A2aR were detected by western blot and immunohistochemical staining techniques. RESULTS: Severe lung fibrosis was observed in the bleomycin-treated mice on day 28. The histopathological findings and collagen content of lung tissues were much severer/higher in A2aR-/- mice than in A2aR+/+ mice. We also showed that TGF-ß1 and p-ERK1/2 were upregulated in bleomycin-treated mice and expressed higher in A2aR-/- mice compared to A2aR+/+ mice. Besides, bleomycin-treated A2aR+/+ mice had increased A2aR level in lungs. However, long-term treatment with baicalin in A2aR-/- and A2aR+/+ mice significantly ameliorated the histopathological changes in lungs. Moreover, Increased TGF-ß1 and p-ERK1/2 expressions in bleomycin-treated A2aR-/- and A2aR+/+ mice were obviously diminished by baicalin. The baicalin-treated A2aR-/- mice had severer lung fibrosis and higher expressions of TGF-ß1 and p-ERK1/2 than A2aR+/+ mice. Baicalin has also upregulated the expression of A2aR in A2aR+/+ mice. CONCLUSIONS: Genetic inactivation of A2aR exacerbated the pathological processes of bleomycin-induced pulmonary fibrosis. Together, baicalin could inhibit BLM-induced pulmonary fibrosis by upregulating A2aR, suggesting A2aR as a therapeutic target of baicalin for the treatment of pulmonary fibrosis.

Effect of arsenite-oxidizing bacterium B. laterosporus on arsenite toxicity and arsenic translocation in rice seedlings.

Arsenite [As (III)] oxidation can be accelerated by bacterial catalysis, but the effects of the accelerated oxidation on arsenic toxicity and translocation in rice plants are poorly understood. Herein we investigated how an arsenite-oxidizing bacterium, namely Brevibacillus laterosporus, influences As (III) toxicity and translocation in rice plants. Rice seedlings of four cultivars, namely Guangyou Ming 118 (GM), Teyou Hang II (TH), Shanyou 63 (SY) and Minghui 63 (MH), inoculated with or without the bacterium were grown hydroponically with As (III) to investigate its effects on arsenic toxicity and translocation in the plants. Percentages of As (III) oxidation in the solutions with the bacterium (100%) were all significantly higher than those without (30-72%). The addition of the bacterium significantly decreased As (III) concentrations in SY root, GM root and shoot, while increased the As (III) concentrations in the shoot of SY, MH and TH and in the root of MH. Furthermore, the As (III) concentrations in the root and shoot of SY were both the lowest among the treatments with the bacterium. On the other hand, its addition significantly alleviated the As (III) toxicity on four rice cultivars. Among the treatments amended with B. laterosporus, the bacterium showed the best remediation on SY seedlings, with respect to the subdued As (III) toxicity and decreased As (III) concentration in its roots. These results indicated that As (III) oxidation accelerated by B. laterosporus could be an effective method to alleviate As (III) toxicity on rice seedlings.

Elastic and hydrostatic behaviour of a zinc dietary supplement, zinc glycinate hydrate.

Coordination polymer based dietary supplement tablets are commonly consumed in our daily life and play an important role in the pharmaceutical industry. To fully understand their tableting process, their mechanical properties need to be comprehensively studied. In this work, the elastic and hydrostatic behaviour of a zinc supplement, zinc glycinate hydrate (Zn[O2CCH2NH2]2·H2O), have been studied via density functional theory (DFT) calculations and high-pressure synchrotron powder X-ray diffraction. This material has a pseudo-layered structure and can be successfully exfoliated into nanosheets. The DFT calculated elastic moduli along the principal axes (13.84-36.11 GPa) indicate a significant elastic anisotropy of ZnG as expected for a layered system, and the directional dependent elastic modulus can be corroborated with the underlying atomic structure. In addition, the calculated B/G ratios (1.30-3.83) according to Pugh's criterion reveal that ZnG could be brittle under uniaxial stress (B and G are bulk modulus and shear modulus, respectively). Furthermore, the measured B is ∼31 GPa, which lies in the middle of the values between inorganic dietary supplements and small organic drug crystals. These results provide some quantitative information about the tableting process of the hybrid dietary supplement which could be different from their inorganic and organic pharmaceutical counterparts.

Expression of autophagy-related factor p62 for lung cancer diagnosis and prognosis: A systematic review and meta-analysis.

p62/SQSTM1 is the scaffold protein implicated in selective autophagy, which is induced by cellular stress. Research has shown that p62 is highly expressed in cancer. Moreover, p62 can easily promote tumor metastasis. However, studies have not reached a consensus on the relationship of p62 expression with the diagnosis and prognosis of lung cancer. We conducted a systematic review and meta-analysis of studies on p62 expression in the prognosis and clinical-pathological parameters of lung cancer patients. Literature search was performed with PubMed, Web of Science, EMBASE, Cochrane Library, and SpringerLink databases. Fixed-effects and random-effects models were used to study the relationship of p62 expression with patients' overall survival (OS) and clinical-pathological parameters. I2 was used to test for heterogeneity. Egger's test was used to assess publication bias. The meta-analysis collected and considered 13 articles, which included 1393 lung cancer patients. The studies show that the high expression of p62 is associated with poor OS in lung cancer patients. The clinical-pathological parameters of patients show that p62 is more highly expressed in high TNM stage (II + III + IV VS. I), Lymph node metastasis (N1 VS. N0), and distant metastases (D1 VS. D0). However, there is no correlation between the p62 expression and the Beclin 1 and LC3B in lung cancer patients. In conclusion, the over-expression of p62 is associated with poor OS in lung cancer patients and can be used as a biomarker for lung cancer diagnosis and prognosis.

Bis{2-[(E)-benzyl-imino-meth-yl]-4,6-dibromo-phenolato-κN,O}cobalt(II).

In the title compound, [Co(C(14)H(10)Br(2)NO)(2)], the Co(II) ion is coordinated by an O and an N atom from two equivalent 2-[(E)-benzyl-imino-meth-yl]-4,6-dibromo-phenolate ligands, displaying a distorted tetra-hedral geometry. The Co(II) ion occupies a special position on a twofold rotation axis and thus the mol-ecular symmetry of the complex is C(2). The two phenolate rings are perpendicular [89.8 (3)°].

Jahn-Teller Effect on Framework Flexibility of Hybrid Organic-Inorganic Perovskites.

Here we study the Jahn-Teller (JT) effect on framework flexibility of two analogous hybrid organic-inorganic perovskites, [C(NH2)3][Zn(HCOO)3] (1-Zn) and [C(NH2)3][Cu(HCOO)3] (2-Cu). Single-crystal nanoindentation measurements show that the elastic moduli and hardnesses of 1-Zn are up to ∼52.0% and ∼25.0% greater than those of the JT active 2-Cu. Temperature-dependent X-ray diffraction measurements indicate that the thermal expansion along the b-axis is switched from negative to positive by replacing Zn2+ with Cu2+ on the B-site. These stark distinctions in framework flexibility are primarily attributed to the ∼10.0% elongation of Cu-O bonds induced by the JT effect and associated alterations in octahedral tilting and hydrogen-bonding. Our results demonstrate the prominence of the JT effect in the emerging hybrid perovskites and highlight the possibilities of tuning materials' properties using orbital order.

A Cd(II)-based metal-organic framework as a luminance sensor to nitrobenzene and Tb(III) ion.

A Cd(II)-based metal-organic framework, [Cd2(DPDC)2(BTB)]∞ (Cd-MOF, DPDC = 2,2'-diphenyldicarboxylate and BTB = 1,4-bis(1,2,4-triazol-1-yl)butane) was successfully constructed via a hydrothermal reaction. Structural analysis shows that the synthesized Cd-MOF is a three-dimensional (3D) architecture crystallized in the hexagonal system with a chiral space group P61. Powder X-ray diffraction experiments and thermogravimetric analysis reveal that the constructed Cd-MOF has a high chemical and thermal stability. A study of additional mechanical properties indicates that it exhibits a moderate stiffness with the average values of Young's modulus (E) and H as 11.3(2) and 0.9(7) GPa, respectively. The luminescence properties of the Cd-MOF were further studied. The result shows that it could be an effective sensor to the organic nitrobenzene molecule via a strong quenching effect, and also to the inorganic Tb(III) ion by a strong green emission effect. Moreover, when loading bimetal ions (Eu(III) and Tb(III) into the Cd-MOF/methanol suspension, tunable visible luminescence can also be achieved by carefully adjusting the excitation wavelengths.

[Study on the interface of human hepatocyte/micropore polypropylene ultrafiltration membrane].

OBJECTIVE: To found a new interface of human hepatocyte/micropore polypropylene ultrafiltration membrane (MPP) with good cytocompatibility so as to construct bioartificial bioreactor with polypropylene hollow fibers in future. METHODS: MPP ultrafiltration membrane underwent chemical grafting modification through ultraviolet irradiation and Fe(2+) reduction. The contact angles of MPP and the modified MPP membranes were measured. Human hepatic cells L-02 were cultured. MPP and modified MPP membranes were spread on the wells of culture plate and human hepatic cells and cytodex 3 were inoculated on them. Different kinds of microscopy were used to observe the morphology of these cells. RESULTS: The water contact angle of MPP and the modified MPP membranes decreased from 78 degrees +/- 5 degrees to 27 degrees +/- 4 degrees (P < 0.05), which indicated that the hydrophilicity of the membrane was improved obviously after the grafting modification. Human hepatocyte L-02 did not adhere to and spread on the modified MPP membrane surface, and only grew on the microcarrier cytodex 3 with higher density and higher proliferation ratio measured by MTT. CONCLUSION: Grafting modification of acrylamide on MPP membrane is a good method to improve the human hepatocyte cytocompatibility with MPP ultrafiltration membrane.

[Retrospective analysis on 682 cases of liver cancer after surgical treatment].

OBJECTIVE: To investigate the factors of long-turn survival of liver cancer after surgical treatment. METHODS: Five hundred and twenty-two cases of liver cancer that received surgical treatment in 14 years were analyzed retrospectively. RESULTS: Comparison between the small liver cancer (< 5 cm) and the greater one (> 10 cm) revealed that the small liver cancer had a higher survival rates than the greater one [3 year (61.25 +/- 4.41)% versus (45.90 +/- 6.98)%; 5 year (53.84 +/- 5.68)% versus (30.21 +/- 10.23)%]. There were same results between single-nodule and two or more than two nodule [3 year (61.86 +/- 3.69)% versus (38.31 +/- 4.97)%; 5 year (55.40 +/- 4.91)% versus (28.01 +/- 6.31)%], between child I and child II or more than II [3 year (60.68 +/- 3.68)% versus (49.88 +/- 4.13)%; 5 year (50.99 +/- 5.10)% versus (36.39 +/- 7.58)%], and between single segmentectomy of the liver and two or more than two segmentectomy [3 year (68.65 +/- 4.95)% versus (49.88 +/- 4.13)%; 5 year (65.38 +/- 5.69)% versus (37.98 +/- 5.70)%]. CONCLUSIONS: Small liver cancer, single-nodule, good hepatic function and minor resection were important factors to prolong survival further.

[Study on the interface of human hepatocyte L-02 polypropylene:simple culture method of human hepatocyte with spheroidal aggregate culture].

OBJECTIVE: To found new interface of human hepatocyte/poly propylene with good cytocompatibility for made polypropylene hollow fibers bioreactor of bioartificial liver in future. METHODS: Using the macromolecular hydroperoxide groups on the polypropylene membrane surface as initiators, acrylamides were polymerized on the polypropylene membranes, under induction by both UV irradiation and Fe2+ reduction. Growth characteristics of human hepatocyte L-02 were detected when it was cultured on polystyrene, polypropylene and modified polypropylene membrane surface. RESULTS: Water contact angle measurement of the polypropylene and the modified polypropylene membranes decreased from (72 +/- 5) degrees to (30 +/- 4) degrees , which indicated that the hydrophilicity of the membrane was improved obviously after the grafting modification. Human hepatocyte L-02 could not adhere and spread on modified polypropylene membrane surface, and grown in spheroidal aggregate with higher density and higher proliferation ratio measured by MTT method. CONCLUSIONS: Acrylamide polymerized on the polypropylene membranes is a good method which not only improved human hepatocytes cytocompatibility but also found a new simple culture method with spheroidal aggregate culture of human hepatocyte.

[Randomized controlled trial of two kinds of home-produced docetaxel in China for advanced breast cancer].

BACKGROUND & OBJECTIVE: Two kinds of home-produced docetaxel in China, injection Yiyoutasai and injection Aisu, have the same structure. Data from preclinical study had shown that injection Yiyoutasai has the same pharmacokinetics and toxicity as injection Aisu. This study was to evaluate the efficacy and toxicity of injection Yiyoutasai in treating advanced breast cancer. METHODS: Eligible breast cancer patients were enrolled and randomly assigned to study group and control group, and received injection of 75 mg/m(2) Yiyoutasai or Aisu, respectively. The injections were repeated every 3 weeks. All patients received at least 2 cycles. The efficacy of Yiyoutasai and Aisu were evaluated after treatment. RESULTS: A total of 67 patients were enrolled: 33 in study group, and 34 in control group. Of the 31 evaluable cases in study group, 1 achieved complete remission (CR), 9 achieved partial remission (PR), 11 had stable disease (SD), and 10 had progressive disease (PD); the total response rate was 22.22%. There were 1 CR, 5 PR, 19 SD, and 9 PD in control group; the total response rate was 15.15%. There was no significant difference between the 2 groups (P=0.662). The median follow-up was 16.5 months (8-28 months). In study group, the median progression-free survival time was 6.2 months (2-12 months), the 1-year survival rate was 68.51%, and the 2-year survival rate was 40.12%; in control group, the median progression-free survival time was 7.1 months (2.3-11 months), the 1-year survival rate was 65.23%, and the 2-year survival rate was 39.71%. There was no significant difference between the 2 groups (P=0.102, 0.098, 0.089, respectively). Common adverse events were myelosuppression, transient transaminase elevation, and alopecia. One patient in study group suffered from severe allergic reaction after infusion, 1 in control group suffered from whole body edema. CONCLUSION: Yiyoutasai and Aisu have similar efficacy on and toxicity to advanced breast cancer patients.

[A randomized control clinical trial of Euruikang (docetaxel) in treatment of advanced non-small cell lung cancer (NSCLC)].

BACKGROUND & OBJECTIVE: Docetaxel is one of the major drugs for advanced non-small cell lung cancer (NSCLC) treatment in clinical use. It is reported that the response rate of docetaxel alone is about 20% and its major toxicity is myelosuppression. The study was to evaluate the efficacy and tolerability of docetaxel (made in Beijing) in the treatment of advanced NSCLC. METHODS: 77 patients were randomized into two groups. In the study group, patients were received docetaxel (made in Beijing) 75 mg/m(2) in one hour plus cisplatin 70 mg/m(2); while in the control group, docetaxel (made in Jiangsu) 75 mg/m(2) plus cisplatin 70 mg/m(2) were administrated. Treatment in the two groups was repeated every 3 weeks. All patients had to receive at least two cycles of chemotherapy. RESULTS: The objective responses in the study and control group were 28.95% and 27.03% respectively, with no statistical difference (P>0.5). Common toxicities of docetaxel (made in Beijing) were grade II-III myelosuppression, grade I-II transaminase elevation, alopecia and hypodynamia. CONCLUSION: Docetaxel (made in Beijing) is an effective chemotherapy drug in advanced NSCLC treatment, which has the similar efficacy and toxicity to docetaxel (made in Jiangsu).