Prevalence and risk factors for subclinical hypothyroidism in older patients with major depressive disorder.

BACKGROUND: Subclinical hypothyroidism (SCH) is highly correlated with major depressive disorder (MDD). However, the prevalence and risk factors for SCH in older patients with MDD have rarely been reported in China. METHODS: This cross-sectional study included 266 older MDD patients with SCH was performed. Clinical and anthropometric, biochemical, and thyroid function data were collected. Depression, anxiety, and psychotic symptoms were assessed using the Hamilton Depression Scale, the Hamilton Anxiety Scale, and the Positive and Negative Syndrome Scale positive subscale, respectively. RESULTS: Among older patients with MDD, the prevalence of SCH was 64.7% (172/266). Compared to patients without SCH, older MDD patients with SCH had a longer disease course and higher TSH, A-TG, A-TPO, HDL-C, LDL-C, TC, FPG, and systolic pressure levels (all P ≤ 0.002). Furthermore, disease progression (OR 1.082, 95% CI 1.020-1.147, P = 0.009), A-TG (OR 1.005, 95% CI 1.001-1.009, P = 0.017), TC (OR 2.024, 95% CI 1.213-3.377, P = 0.007), FPG (OR 2.916, 95% CI 1.637-5.194, P < 0.001), systolic pressure (OR 1.053, 95% CI 1.008-1.100, P = 0.022) were independently associated with SCH, in older patients with MDD. CONCLUSIONS: Our findings suggest a high prevalence of SCH in older patients with MDD. Several demographic and clinical variables were independently associated with SCH in older patients with MDD.

[Mechanism of Mailuo Shutong Pills on posterior limb muscle swelling caused by femoral fracture in rats based on network pharmacology and experimental verification].

This study aims to investigate the mechanism of Mailuo Shutong Pills(MLST) on posterior limb muscle swelling caused by femoral fracture(SCFF) through network pharmacology and animal experiments. The plasma components of MLST were analyzed by LC-MS, and the target and signal pathway of SCFF were predicted by network pharmacology and verified by molecular docking. SCFF model rats were established through animal experiments, and different doses of MLST were administered to detect the degree of limb swelling. Hematoxylin-eosin(HE) staining was used to observe pathological changes in muscle tissue, and interleukin-6(IL-6), interleukin-1ß(interleukin-1ß), and tumor necrosis factor-α(TNF-α) in peripheral blood were determined by enzyme-linked immunosorbent assay(ELISA). The expression of relevant signaling pathways was measured by Western blot. Network pharmacological results showed that MLST and SCFF had a total of 153 disease targets, and the key targets were IL-6, TNF, etc., involving mitogen-activated protein kinase(MAPK) signaling pathway, phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT) signaling pathway, etc. The binding energies of the main components and key targets were lower than-7.0 kcal·mol~(-1), indicating that the network analysis results were reliable. The results of animal experiments showed that MLST could reduce the swelling degree and pathological damage of the posterior limb muscles of SCFF rats compared with the model group. ELISA results showed that MLST could reduce the levels of IL-6, IL-1ß, and TNF-α in the serum of SCFF rats. Western blot results showed that MLST can reduce the expression of p-AKT, p-PI3K, p-NF-κB, p-p38 MAPK, and p-ERK in SCFF rats. MLST may reduce the content of inflammatory factors in serum by regulating the expression of PI3K/AKT and MAPK-related signaling pathway protein and improving posterior limb muscle SCFF in rats.

[Establishment of quality evaluation methods for pieces and standard decoction of honey-fried Descurainiae Semen].

To establish a content determination method for quality control of the pieces and standard decoction of honey-fried Descurainiae Semen. Standard decoction of honey-fried Descurainiae Semen was prepared with standardized process, and high performance liquid chromatography coupled with diode-array detector(HPLC-DAD) was used to detect its characteristic fingerprint and determine the content of quercetin-3-O-ß-D-glucose-7-O-ß-D-gentiobioside. In addition, the transfer rate, dry extract rate and pH value were calculated. The results showed that the established method had a high accuracy. The content of quercetin-3-O-ß-D-glucose-7-O-ß-D-gentiobioside in 13 batches of standard decoction was 0.03-0.12 mg·mL~(-1); the transfer rate was 13.4%-23.1%; the rate of extracts was 1.9%-5.5%, and the pH was between 5.4-5.9. The similarity coefficients were all greater than 0.85, indicating good homogeneity for the different batches of decoction. There were 7 common peaks in the characteristic chromatogram, one of which was quercetin-3-O-ß-D-glucose-7-O-ß-D-gentiobioside. In this paper, the established content determination and quality evaluation method for Descurainiae Semen pieces and decoction was simple, rapid and reproducible, providing reference for the quality control of honey-fried Descurainiae Semen pieces, standard decoction and its preparations.

Integrated Exploitation of the Structural Diversity Space of Chemotherapy Drugs to Selectively Inhibit HER2 T798M Mutant in Lung Cancer.

An acquired T798M gatekeeper mutation in human epidermal growth factor receptor 2 (HER2) kinase can cause drug resistance to anti-HER2 chemotherapy drugs in lung cancer. Previously, the reversible pan-kinase inhibitor staurosporine has been found to selectively inhibit the HER2 T798M mutant over wild-type kinase, suggesting that the staurosporine scaffold is potentially to develop mutant-selective inhibitors. Here, we systematically evaluated the chemical space of staurosporine scaffold-based compounds in response to HER2 T798M mutation at structural, energetic and molecular levels by using an integrated analysis strategy. With this strategy, we were able to identify several novel wild-type sparing inhibitors with high or moderate selectivity, which are comparable to or even better than that of the parent compound staurosporine. Molecular modeling and structural analysis revealed that noncovalent contacts can form between the side chain of mutated residue Met798 and selective inhibitor ligands, which may improve the favorable interaction energy between the kinase and inhibitor and reduce the unfavorable desolvation penalty upon the kinase-inhibitor binding.

Fluorescence-Enhanced Sensing of Hypochlorous Acid Based on 2-Pyridylthiazole Unit.

Hypochlorous acid, being one of reactive oxygen species (ROS), is essential to protect the body against invasion of pathogens. Excess of hypochlorous acid (HOCl) is believed to be in tight connection with various inflammation-related diseases. It remains a challenge to detect the ROS in physiological conditions (aqueous buffer and neutral pH) with selectivity. In the presented paper, we have synthesized a ferrocence-modified pyridylthiazole derivatives, 1,4-di{5-[(4'-ferrocenyl-2'-(4"-pyridyl)]thiazinyl}benzene (DFPT). Only HOCl could turn-on the fluorescence of DFPT with enhanced emission at 465 nm. Compared to the other reported HOCl sensors, DFPT could selectively detect HOCl with rapid response (< 60 s) in the aqueous buffer (pH = 7.0). The detection limit at pH = 7.0 was 0.7 µM according to the titration experiment.

Interleukin-17A antagonist attenuates radiation-induced lung injuries in mice.

OBJECTIVE: To investigate the effect of interleukin-17A (IL-17A) antibodies on radiation-induced lung injuries in mice. METHODS: The thorax of 135 mice were divided into Sham (n = 30), radiation control (RC, n = 35), treatment (n = 35, IL-17A-neutralizing antibody, 4 µg/mouse, IV, 4 days per month for 4 months) and placebo group (n = 35) before a single dose irradiation (15 Gy) to the thorax. Inflammation and collagen contents in the lung tissues were examined, and the concentration of IL-17A, TGF-ß1, and IL-6 in bronchoalveolar lavage fluid (BALF) were measured. In another 50 animals, 180-day survival rate following the irradiation and treatment was calculated by Kaplan-Meier method. RESULTS: Sixteen weeks after the irradiation and treatment, there was significant inflammatory cell infiltration and interstitial collagen depositions in the radiation control and placebo groups, whereas these changes were relatively mild in the treatment group. The percentage of grade II and III alveolitis in the treatment group (16%, P < .05) was lower than in the RC (72%) or placebo group (64%). The mean Aschcroft fibrosis scores were 2.8 (treatment group), 5.2 (RC), and 4.8 (placebo group), respectively. The scores of treatment group was lower than that of RC (P < .001) or placebo group (P < .001). The IL-17A, TGF-ß, and IL-6 concentrations in the treatment group were lower than in the RC and placebo group (P < .01) following the irradiation. The 180-day mortality rate in the treatment group was lower than in the RC group 16.7% versus 75.0%, P < .05). CONCLUSION: IL-17A antibody treatment alleviates radiation-induced pneumonitis and subsequent fibrosis, and improvise postirradiation survival.

Expression of angiogenesis regulatory proteins and epithelial-mesenchymal transition factors in platelets of the breast cancer patients.

Platelets play a role in tumor angiogenesis and growth and are the main transporters of several angiogenesis regulators. Here, we aimed to determine the levels of angiogenesis regulators and epithelial-mesenchymal transition factors sequestered by circulating platelets in breast cancer patients and age-matched healthy controls. Platelet pellets (PP) and platelet-poor plasma (PPP) were collected by routine protocols. Vascular endothelial growth factor (VEGF), platelet-derived growth factor BB (PDGF-BB), thrombospondin-1 (TSP-1), platelet factor 4 (PF4), and transforming growth factor-ß1 (TGF-ß1) were measured by enzyme-linked immunosorbent assay. Angiogenesis-associated expression of VEGF (2.1 pg/10(6) platelets versus 0.9 pg/10(6) platelets, P < 0.001), PF4 (21.2 ng/10(6) platelets versus 10.2 ng/10(6) platelets, P < 0.001), PDGF-BB (42.9 pg/10(6) platelets versus 19.1 pg/10(6) platelets, P < 0.001), and TGF-ß1 (15.3 ng/10(6) platelets versus 4.3 ng/10(6) platelets, P < 0.001) differed in the PP samples of cancer and control subjects. In addition, protein concentrations were associated with clinical characteristics (P < 0.05). Circulating platelets in breast cancer sequester higher levels of PF4, VEGF, PDGF-BB, and TGF-ß1, suggesting a possible target for early diagnosis. VEGF, PDGF, and TGF-ß1 concentrations in platelets may be associated with prognosis.

Pegylated filgrastim is comparable with filgrastim as support for commonly used chemotherapy regimens: a multicenter, randomized, crossover phase 3 study.

The purpose of this study was to compare the efficacy and safety of a single subcutaneous injection of pegylated filgrastim with daily filgrastim as a prophylaxis for neutropenia induced by commonly used chemotherapy regimens. Fifteen centers enrolled 337 chemotherapy-naive cancer patients with normal bone marrow function. All patients randomized into AOB and BOA arms received two cycles of chemotherapy. Patients received a single dose of pegylated filgrastim 100 µg/kg in cycle 1 (AOB) or cycle 2 (BOA) and daily doses of filgrastim 5 µg/kg/day in cycle 1 (BOA) or cycle 2 (AOB). Efficacy and safety parameters were recorded. The primary end point was the rate of protection against grade 4 neutropenia after chemotherapy [defined as the rate at which the absolute neutrophil count (ANC) remained >0.5×10(9)/l throughout the entire cycle]. Ninety-four percent of patients receiving pegylated filgrastim or filgrastim did not develop grade 4 neutropenia. The incidence of ANC<1.0×10(9)/l was 16.0% (50/313) after support with either pegylated filgrastim or filgrastim. The incidences of febrile neutropenia and antibiotic administration were similar in both groups. Notably, faster ANC recovery was observed with pegylated filgrastim support. The ANC nadir was also earlier with pegylated filgrastim (day 7) support than with filgrastim support (day 9), although the depth of nadir was not significantly different. A single subcutaneous injection of pegylated filgrastim 100 µg/kg provided adequate and safe neutrophil support comparable with daily subcutaneous injections of unmodified filgrastim 5 µg/kg/day in patients receiving commonly used standard-dose mild-to-moderate myelosuppressive chemotherapy regimens.

Promoter methylation of tumor suppressor genes in esophageal squamous cell carcinoma.

Esophageal squamous cell carcinoma (ESCC) is a prevalent and fatal cancer in China and other Asian countries. Epigenetic silencing of key tumor suppressor genes (TSGs) is critical to ESCC initiation and progression. Recently, many novel TSGs silenced by promoter methylation have been identified in ESCC, and these genes further serve as potential tumor markers for high-risk group stratification, early detection, and prognosis prediction. This review summarizes recent discoveries on aberrant promoter methylation of TSGs in ESCC, providing better understanding of the role of disrupted epigenetic regulation in tumorigenesis and insight into diagnostic and prognostic biomarkers for this malignancy.

E-cadherin promotes proliferation of human ovarian cancer cells in vitro via activating MEK/ERK pathway.

AIM: E-cadherin is unusually highly expressed in most ovarian cancers. This study was designed to investigate the roles of E-cadherin in the carcinogenesis and progression of ovarian cancers. METHODS: Human ovarian adenocarcinoma cell line SKOV-3 was examined. E-cadherin gene CDH1 in SKOV-3 cells was knocked down via RNA interference (RNAi), and the resultant variation of biological behavior was observed using CCK-8 and colony formation experiment. E-cadherin-mediated Ca(2+)-dependent cell-cell adhesion was used to study the mechanisms underlying the effects of E-cadherin on the proliferation and survival of SKOV-3 cells. The expression levels of E-cadherin, extracellular signal-related kinase (ERK), phosphorylated ERK (P-ERK) were measured using Western blot assays. RESULTS: Transfection with CDH1-siRNA for 24-96 h significantly suppressed the growth and proliferation of SKOV-3 cells. E-cadherin-mediated calcium-dependent cell-cell adhesion of SKOV-3 cells resulted in a rapid increase of P-ERK, but did not modify the expression of ERK protein. The phosphorylation of ERK in the cells was blocked by pretreatment with the MEK1 specific inhibitor PD98059 (50 µmol/L), but not by the PI3K inhibitor wortmannin (1 µmol/L) or PKA inhibitor H89 (10 µmol/L). CONCLUSION: E-cadherin may function as a tumor proliferation enhancer via activating the MEK/ERK pathway in development of ovarian epithelial cancers.

FG020326 sensitized multidrug resistant cancer cells to docetaxel-mediated apoptosis via enhancement of caspases activation.

Apoptotic resistance is the main obstacle for treating cancer patients with chemotherapeutic drugs. Multidrug resistance (MDR) is often characterized by the expression of P-glycoprotein (P-gp), a 170-KD ATP-dependent drug efflux protein. Functional P-gp can confer resistance to activate caspase-8 and -3 dependent apoptosis induced by a range of different stimuli, including tumor necrosis and chemotherapeutic drugs such as docetaxel and vincristine. We demonstrated here that comparison of sensitive KB cells, P-gp positive (P-gp(+ve)) KBv200 cells were extremely resistant to apoptosis induced by docetaxel. FG020326, a pharmacological inhibitor of P-gp function, could enhance concentration-dependently the effect of docetaxel on cell apoptosis and sensitize caspase-8, -9 and -3 activation in P-gp overexpressing KBv200 cells, but not in KB cells. Therefore, the enhancement of caspase-8, -9 and -3 activation induced by docetaxel may be one of the key mechanisms of the reversal of P-gp mediated docetaxel resistance by FG020326.

[Promoting of angiogenesis and osteogenesis in radial critical bone defect regions of rabbits with nano-hydroxyapatite/collagen/PLA scaffolds plus endothelial progenitor cells].

OBJECTIVE: To explore the roles of nano-hydroxyapatite/collagen/PLA (nHAC/PLA) plus endothelial progenitor cells (EPCs) in repairing segmental bone defects of rabbit radius and enhancing angiogenesis and new bone formation. METHODS: EPCs isolated from New Zealand white rabbit bone marrow were cultured, identified and seeded into nHAC/PLA scaffolds. And the growth of EPCs in scaffolds was observed under scanning electron microscopy (SEM). Thirty-six were randomly divided into 3 groups to establish segmental bone defect models in radii. Two groups were implanted with EPCs/scaffolds constructs (group A, n = 16) and scaffolds alone (group B, n = 16) respectively. The remaining four rabbits were used as negative control (group C) and nothing was implanted. Animals were sacrificed at different timepoints and radii harvested to undergo radiological examination, histological examination and microvessle density test. RESULTS: These cells isolated from bone marrow were confirmed as EPCs. SEM showed that EPCs attached to the nHAC/PLA scaffolds, grew and proliferated well. Animal experiments revealed that radiological scores (5w: 2.25 ± 0.50 vs 1.00 ± 0.00; 10w: 2.75 ± 0.50 vs 1.75 ± 0.50; 15w: 4.25 ± 0.50 vs 3.0 ± 0.0; each P < 0.05), percentage of new bone formation area in bone defect regions (5w: 29.0% ± 3.5% vs 8.1% ± 0.8%; 10w: 63.4% ± 5.5% vs 16.6% ± 1.3%; 15w: 96.0% ± 4.3% vs 34.0% ± 6.6%; each P < 0.05) and microvessel density (2w: 13.5 ± 0.9 vs 4.3 ± 1.0; 5w:9.8 ± 0.7 vs 4.8 ± 0.3; 10w: 7.0 ± 0.4 vs 4.5 ± 0.4; each P < 0.05) in group A were significantly higher than those in group B. No new bone formation occurred in group C. CONCLUSION: The composite structure of EPCs-nHAC/PLA can enhance angiogenesis and new bone formation in segmental bone defects in rabbit radii. It may become a potential candidate of promoting revascularization of tissue engineering bone and repairing large bone defects.

CCL5-Secreting Virtual Memory CD8+ T Cells Inversely Associate With Viral Reservoir Size in HIV-1-Infected Individuals on Antiretroviral Therapy.

Recent studies highlighted that CD8+ T cells are necessary for restraining reservoir in HIV-1-infected individuals who undergo antiretroviral therapy (ART), whereas the underlying cellular and molecular mechanisms remain largely unknown. Here, we enrolled 60 virologically suppressed HIV-1-infected individuals, to assess the correlations of the effector molecules and phenotypic subsets of CD8+ T cells with HIV-1 DNA and cell-associated unspliced RNA (CA usRNA). We found that the levels of HIV-1 DNA and usRNA correlated positively with the percentage of CCL4+CCL5- CD8+ central memory cells (TCM) while negatively with CCL4-CCL5+ CD8+ terminally differentiated effector memory cells (TEMRA). Moreover, a virtual memory CD8+ T cell (TVM) subset was enriched in CCL4-CCL5+ TEMRA cells and phenotypically distinctive from CCL4+ TCM subset, supported by single-cell RNA-Seq data. Specifically, TVM cells showed superior cytotoxicity potentially driven by T-bet and RUNX3, while CCL4+ TCM subset displayed a suppressive phenotype dominated by JUNB and CREM. In viral inhibition assays, TVM cells inhibited HIV-1 reactivation more effectively than non-TVM CD8+ T cells, which was dependent on CCL5 secretion. Our study highlights CCL5-secreting TVM cells subset as a potential determinant of HIV-1 reservoir size. This might be helpful to design CD8+ T cell-based therapeutic strategies for cure of the disease.

Epimeric monosaccharide-quinone hybrids on gold electrodes toward the electrochemical probing of specific carbohydrate-protein recognitions.

Carbohydrates represent one of the most significant natural building blocks, which govern numerous critical biological and pathological processes through specific carbohydrate-receptor interactions on the cell surface. We present here a new class of electrochemical probes based on gold surface-coated epimeric monosaccharide-quinone hybrids toward the ingenious detection of specific epimeric carbohydrate-protein interactions. Glucose and galactose, which represent a pair of natural monosaccharide C4 epimers, were used to closely and solidly conjugate with the 1,4-dimethoxybenzene moiety via a single C-C glycosidic bond, followed by the introduction of a sulfhydryl anchor. The functionalized aryl C-glycosides were sequentially coated on the gold electrode via the self-assembled monolayer (SAM) technique. X-ray photoelectron spectroscopy (XPS) was used to confirm the SAM formation, by which different binding energies (BE) between the glucosyl and the galactosyl SAMs on the surface, probably rendered by their epimeric identity, were observed. The subsequent electrochemical deprotection process readily furnished the surface-confined quinone/hydroquinone redox couple, leading to the formation of electrochemically active epimeric monosaccharide-quinone SAMs on the gold electrode. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) used for the detection of specific sugar-lectin interactions indicated that the addition of specific lectin to the corresponding monosaccharide-quinone surface, i.e., concanavalin A (Con A) to the glucosyl SAM and peanut agglutinin (PNA) to the galactosyl SAM, resulted in an obvious decrease in peak current, whereas the addition of nonspecific lectins to the same SAMs gave very minor current variations. Such data suggested our uniquely constructed gold surface coated by sugar-quinone hybrids to be applicable as electrochemical probes for the detection of specific sugar-protein interactions, presumably leading to a new electrochemistry platform toward the study of carbohydrate-mediated intercellular recognitions.

[Influence of minor back trauma on surgical outcomes in patients with thoracic ossification of ligamentum flavum].

OBJECTIVE: To retrospectively explore the influences of minor back trauma on surgical outcomes in patients with thoracic ossification of ligamentum flavum (TOLF) and preliminarily detect its possible causes. METHODS: A total of 94 TOLF patients were divided into two groups according to the absence or presence of minor back trauma: MT (minor trauma, n = 16) and NT (no trauma, n = 78). They were compared in terms of gender, age, duration of symptoms, levels of involvement, numbers of involved segments, ratio of intramedullary signal changes (IMSC), pre- & post-operative JOA (Japanese Orthopedic Association) score, recovery rate (RR) at the final follow-up. Multiple regression analysis was employed to elucidate the causes related with the surgical outcomes. The MT group was further divided into two subgroups according to the intervals between trauma and surgery to clarify the influences of surgical timing on the efficacies. RESULTS: The JOA scores were 4.0 ± 1.4 and 8.4 ± 1.7 respectively in MT and NT groups at the final follow-up. The neurological status of patients improved in both groups (MT: P = 0.009, NT: P = 0.000). The patients were younger in MT groups (50 ± 11 years) than those in NT groups (58 ± 8 years) (P = 0.046). The ratio of IMSC was higher in MT groups (75.0%) than that in NT groups (25.6%) (P = 0.000). The pre- & post-operative JOA scores were lower in MT groups than those in NT groups (both P = 0.000). Multiple regression analysis revealed that the postoperative JOA score at the final follow-up was positively related with the preoperative JOA score (r = 0.60, P = 0.000) and negatively with trauma and IMSC (r = -1.82 and r = -1.87, P = 0.000) while the final postoperative RR were negatively related with trauma and IMSC (r = -26.26 and r = -33.70, P = 0.000). The surgical timing after trauma did not influence the efficacies (P = 0.147). CONCLUSION: The TOLF patients with minor back trauma have a worse post-operative recovery. A minor trauma might be a risk factor of adverse surgical outcomes.

Changes of Damage Associated Molecular Patterns in COVID-19 Patients.

Background: The development of severe coronavirus disease 2019 (COVID-19) is associated with systemic hyperinflammation, which drives multi-organ failure and death. Disease deterioration tends to occur when the virus is receding; however, whether other factors besides viral products are involved in the inflammatory cascade remains unclear. Methods: Twenty-eight COVID-19 patients with laboratory-confirmed SARS-CoV-2 infection hospitalized at the Fifth Medical Center of Chinese PLA General Hospital from January 23 to February 20, 2020 and nine healthy donors during the same period were recruited in the study. COVID-19 patients were grouped as mild, moderate, severe based on disease severity. Plasma damage-associated molecular patterns (DAMPs), including high mobility group box 1 (HMGB1), calprotectin (S100A8/A9), surfactant protein A (SP-A), cold-inducible RNA-binding protein (CIRBP), and Histone H4 were detected by ELISA assay, and analyzed in combination with clinical data. Plasma cytokines, chemokines and lymphocytes were determined by flow cytometry. Results: Plasma levels of HMGB1 (38292.3 ±â€Š4564.4 vs. 32686.3 ±â€Š3678.1, P = 0.002), S100A8/A9 (1490.8 ±â€Š819.3 vs. 742.2 ±â€Š300.8, P = 0.015), and SP-A (6713.6 ±â€Š1708.7 vs. 5296.3 ±â€Š1240.4, P = 0.048) were increased in COVID-19 patients compared to healthy donors, while CIRBP (57.4 ±â€Š30.7 vs. 111.9 ±â€Š55.2, P = 0.004) levels decreased. Five DAMPs did not vary among mild, moderate, and severe patients. Moreover, SP-A levels correlated positively with inflammatory cytokines and negatively with time elapsed after symptom onset, whereas CIRBP showed an opposite pattern. Conclusions: These findings suggest SP-A may involve in the inflammation of COVID-19, while CIRBP likely plays a protective role. Therefore, DAMPs represent a potential target in the prevention or treatment of COVID-19.

[Effect of siRNA-mediated beta-catenin gene on Wnt signal pathway in lung adenocarcinoma A549 cell].

OBJECTIVE: To study the effects of siRNA-mediated beta-catenin down-regulation on Wnt/beta-catenin signal transduction pathway in lung adenocarcinoma A549 cells. METHODS: A549 cell was divided into three groups: PGCsil-CTNNB1-siRNA group (transfection with beta-catenin interference plasmids), negative control group (transfection with negative control plasmids) and non-transfection group (blank control). For each group, real-time PCR was employed to detect the expression of beta-catenin and cyclin D1 (a target gene of Wnt/beta-catenin pathway). A MTT cell proliferation assay was performed to study the proliferating capacity. Flow cytometry was used for cell cycle analysis. Clone formation and Millicell chamber experiment were performed to evaluate the clone formation and migration capacities of cells. RESULTS: The expression of beta-catenin and cyclin D1 in PGCsil-CTNNB1-siRNA cell decreased at the mRNA level. It was lower than negative control (0.002+/-0.001 vs 0.023+/-0.002, P<0.01; 0.005+/-0.002 vs 0.040+/-0.020, P<0.05) and blank control groups (beta-catenin mRNA: 0.021+/-0.003, P<0.01; cyclin D1 mRNA: 0.042+/-0.004, P<0.05). Also the proliferating capacity at Days 5-7 was inhibited in comparison with negative control and blank control groups (P<0.05, P<0.01). The cell-doubling time (58.1 h) was markedly longer than those of negative control group (37.9 h, P<0.05) and blank control group (34.2 h, P<0.05). The number of cells in G0-G1 phrase increased in PGCsil-CTNNB1-siRNA group (86.4%+/-2.6%) in comparison with negative control (73.8%+/-0.9%, P<0.01) and blank control groups (75.8%+/-1.5%, P<0.01). In PGCsil-CTNNB1-siRNA group, the clone formation ratio (31.6%+/-7.7%) was lower than negative control (46.9%+/-7.3%, P<0.05) and blank control groups (44.2%+/-2.5%, P<0.05). And the number of cells (16.0+/-3.8) passing through the membrane of Millicell chamber was smaller than negative control (32.7+/-3.1, P<0.01) and blank control groups (33.0+/-2.7, P<0.01). CONCLUSIONS: Knocking down beta-catenin not only inhibits the Wnt/beta-catenin signal transduction pathway in lung adenocarcinoma A549 cells but also decreases cell proliferation, clone formation and migration capacity. Thus it may become a novel target for lung cancer therapy.

Three pairs of luminescent coordination polymers based on CoII and CdII clusters for the detection of antibiotics, pesticides and chiral nitro aromatic compounds.

Three couples of coordination polymers (CPs), namely, [Co((R/S)-Hcna)2] n (1-D/L), [Cd6((1R,2R/1S,2S)-cpba)4(phen)6(H2O)3] n (2-D/L) and [Cd2((1R,2R/1S,2S)-Hcpba)2(phen)2] n (3-D/L) {(R/S)-H2cna = (R/S)-6-(1-carboxyethoxy)-2-naphthoic acid, (1R,2R/1S,2S)-H3cpba = (1R,2R/1S,2S)-2,2'-((5-carboxy-1,3-phenylene)bis(oxy))dipropionic acid, phen = 1,10-phenanthroline} are successfully synthesized under hydrothermal conditions. Structural analysis shows that CP 1 has a 3D 3,6-c net structure with a point symbol of (4·62)2(42·610·83). CPs 2 and 3 are obtained under very similar reaction conditions except using different solvent ratios. The presence of the planar chelating ligand phen in CPs 2 and 3 limited the spatial growth of the structure, resulting in the formation of different 1D structures. All CPs crystallized in the chiral space group P21, CPs 1-3 are all SHG active. Their luminescence sensing activities for organics such as antibiotics, pesticides and nitro aromatics are also investigated. The results showed that CP 1 can effectively identify trace amounts of nitrofurans (NFs) and CP 3 has obvious recognition ability toward nitrofurans (NFs) and nitroimidazoles (NMs). Both CPs 1 and 3 could selectively detect 2,6-dichloro-4-nitroaniline (DCN). The luminescence of CPs 1 and 3 can also be quenched by (D/L)-4-nitrophenylalanine ((D/L)-NPA) and (1R,2R/1S,2S)-2-amino-1-(4-nitrophenyl)propane-1,3-diol ((1R,2R/1S,2S)-ANPO).

Detection of four phenolic oestrogens by a novel electrochemical immunosensor based on a hexestrol monoclonal antibody.

Using a sandwich-type immunoassay, a novel electrochemical immunosensor based on the successful preparation of a hexestrol (HEX) monoclonal antibody was successfully constructed to detect four phenolic oestrogens: HEX, diethylstilboestrol (DES), dienestrol (DE) and bisphenol A (BPA). An innovative strategy to design the HEX monoclonal antibody/mercapto acetic acid (MACA)/nanogold immunosensor was developed by combining a nanosized effect, self-assembly and specific immune technology. The differential pulse voltammetry (DPV) response for four phenolic oestrogens was in the order BPA > DE > HEX > DES with detection limits of 0.037, 0.047, 0.052 and 0.060 ng mL-1, respectively (S/N = 3). This immunosensor exhibits superior electrochemical properties with a wide linear range, low detection limit, excellent reproducibility and high selectivity. Compared with a high-performance liquid chromatography method, a high accuracy of DES detection in beef, duck and milk powder samples was achieved by the immunosensor proposed, which exhibits great potential for the detection of phenolic oestrogens in applications in environmental and food safety.

[Phase III clinical study of zoledronic acid in the treatment of pain induced by bone metastasis from solid tumor or multiple myeloma].

OBJECTIVE: To evaluate the efficacy and safety of zoledronic acid in the treatment of bone pain in patients with bone metastasis from solid tumor or multiple myeloma. METHODS: A randomized, double-blind, double-simulated and multi-center phase III clinical trail with pamidronate as control was conducted. Patients with moderate to severe bone pain (VAS > 50 mm) induced by solid tumor or multiple myeloma were randomized to receive intravenous zoledronic acid 4 mg or pamidronate 90 mg. Then the change of VAS and urinary NTX/Cr and CTX/Cr were observed in two groups. RESULTS: From July 2005 to September 2006, 228 patients with bone pain induced by bone metastasis from 15 cancer centers were randomize into two groups: 116 patients in zoledronic acid group and 112 patients in pamidronate group. The VAS value was decreased gradually after treatment in these two groups. Significant improvement in bone pain after treatment were observed both in zoledronic acid group and the control group when compared with baseline VAS on D8 (-11.77% vs. -10.87%), D15 (-24.60% vs. -21.06%) and D28 (-32.37% vs. -31.26%) (P< or =0.0001), but no significant difference existed between two groups (P =0.6587). Compared with baseline, urine NTX/Cr and CTX/Cr level were decreased rapidly after treatment in both groups, the nadir was on D8, the median decreased on D28, which was -36.9% vs. -32.1% for NTX/Cr (P = 0.7922) and -63.2% vs. -47.9% for CTX/Cr (P =0.834). The frequently observed adverse events were pyrexia (19.0% vs. 31.3%), vomiting (6.0% vs. 8.9%), nausea (4.3% vs. 4.5%), fatigue (3.4% vs. 2.7%) and constipation (2.6% vs. 1.8%) in the two groups. Compared with baseline, the serum creatinine level was not significantly increased throughout the study. CONCLUSION: Intravenous injection of 4 mg zoledronic acid can significantly reduce bone pain and bone resorption marker in urine in the Chinese patients with bone metastasis from solid tumor or multiple myeloma, which is tolerable and also comparable to pamidronate in the efficacy and safety.