A novel synthetic compound shows antioxidant and anti-inflammatory activity and alleviates cognitive deficits in rats for the treatment of Alzheimer's disease.

BACKGROUND: Alzheimer's disease (AD) is a progressive dementia, and ß-amyloid (Aß) accumulation is widely regarded as the primary pathogenesis of AD. A new synthetic compound, 8-hydroxyquinoline-resveratrol derivative (E)-5-(4-hydroxystyryl)quinolin-8-ol (10c) was evaluated as a possible anti-AD agent. METHOD: (I) The total amount of ROS in SH-SY5Y cells was detected by dichlorofluorescein diacetate (DCFH-DA), and the antioxidant activity and neuroprotective effect of 10c in SH-SY5Y cells were evaluated; (II) Griess reagent was used to test the activity of Compound 10c against NO production in LPS-induced BV-2 microglial cells; (III) An automatic digital stereotaxic instrument was used to inject Aß25-35 into the brain to establish an AD animal model to evaluate the protective effect of compound 10c on Aß25-35-induced learning and memory dysfunction in rats. RESULTS: 10c exhibited far more potent antioxidant activity for both exogenous and endogenous reactive oxygen species (ROS) than trolox, resveratrol, and CQ (ROS production: 10c with 26.23% at 1.5 µM; resveratrol with 82.17% at 2.5 µM; CQ with 78.52% at 10 µM). 10c also shows good neuroprotective effects as an endogenous antioxidant in neuroblastoma cells. Moreover, Compound 10c also demonstrated effective inhibition of nitric oxide (NO) production (IC50 =3.10 µM) and IL-1ß production in BV-2 microglial cells which were treated with lipopolysaccharide (LPS). In the water maze test, the numbers of rats who crossed the former platform were increased significantly in both the 10c group (5.7±1.6) and positive control group (CQ, 5.1±1.7). Meanwhile, both 10c (43.8±5.5 s) and CQ (44.1±6.6 s) treatment could significantly prolong the time rats spent in the target quadrant compared to the vehicle-treated model group. These results demonstrated that 10c could alleviate the learning and memory dysfunction of rats induced by Aß25-35 to a certain extent. CONCLUSIONS: Altogether, compound 10c is a promising compound for the treatment of AD.

[Effect and Involved Mechanism of RSL3-induced Ferroptosis in Acute Leukemia Cells MOLM13 and Drug-resistant Cell Lines].

OBJECTIVE: To investigate the effect and involved mechanism of RSL3 on ferroptosis action in acute leukemia cells MOLM13 and its drug-resistant cells. METHODS: After MOLM13 treated with RSL3, CCK-8 assay was performed to detect cell viability, flow cytometry was used to detect the reactive oxygen species (ROS) level of the cells, RT-qPCR and Western blot were used to detect the expression of glutathione peroxidase 4 (GPX4). After MOLM13/IDA and MOLM13/Ara-C, the drug-resistant cell lines were constructed, the ferroptosis induced by RSL3 was observed. Bone marrow samples were collected from patients with acute monocytic leukemia. RT-qPCR and Western blot were performed to detect the expression of related genes and proteins involved in ferroptosis pathway. RESULTS: RSL3 significantly inhibited the cell viability of MOLM13 and increased the intracellular ROS level, which were partially reversed by ferrostatin-1. The mRNA and protein expression of GPX4 decreased in MOLM13 treated with RSL3. RSL3 inhibited the viability of MOLM13/IDA and MOLM13/Ara-C cells more strongly than that of non-drug resistant cells, also increased the intracellular ROS level . The cytotoxic effects were partially reversed by ferrostatin-1. The mRNA and protein expressions of GPX4 in MOLM13/IDA and MOLM13/Ara-C cells were higher than those in non-drug resistant cells. The mRNA and protein levels of GPX4 in bone marrow of relapsed/refractory acute mononuclear leukemia patients were higher than those of ordinary acute mononuclear leukemia patients. CONCLUSION: RSL3 can induce non-drug resistant cells MOLM13 ferroptosis by inhibiting GPX4 activity. MOLM13/IDA and MOLM13/Ara-C are more sensitive to RSL3 compared with non-drug resistant cells MOLM13, which may be caused by the differences in GPX4 expression. The expressions of GPX4 mRNA and protein in relapsed/refractory acute mononuclear leukemia are higher than those in ordinary acute mononuclear leukemia.

Efficacy of transcatheter arterial chemoembolization combined with capecitabine and cetuximab in the treatment of colorectal cancer with liver metastasis.

PURPOSE: The purpose of this study was to investigate the efficacy and safety of transcatheter arterial chemoembolization (TACE) combined with capecitabine and cetuximab in the treatment of colorectal cancer with liver metastasis. METHODS: The colorectal cancer patients with liver metastasis were divided into two groups, namely, Capecitabine group (receiving TACE combined with capecitabine and cetuximab, n=70) and Control group (undergoing TACE combined with cetuximab, n=70). The short-term clinical efficacy, serum tumor markers and liver function indexes were compared. Besides, the survival of patients was analyzed. RESULTS: At 3 months after treatment, the serum levels of carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9) and vascular endothelial growth factor (VEGF) were significantly lower than those before treatment in both group, and they were lower in Capecitabine group than those in Control group after treatment. The liver function indexes, alanine aminotransferase and aspartate aminotransferase, were significantly increased, while the level of albumin was significantly decreased in both groups at 3 d after treatment, and they were improved significantly at 7 d after treatment in contrast with those at 3 d after treatment. After treatment, there were no statistically significant differences in the Karnofsky performance status score and Quality of Life score between Capecitabine group and Control group. The median survival time of patients in Capecitabine group and Control group was 18.1 months and 14.7 months, respectively. There was a statistically significant difference in the 1-year overall survival rate between Capecitabine group and Control group. Moreover, the cumulative survival rate was significantly higher in Capecitabine group than that in Control group. CONCLUSION: The short-term efficacy of TACE combined with capecitabine and cetuximab in treating colorectal cancer with liver metastasis is superior to that of TACE combined with cetuximab.

Safety and effectiveness of argatroban versus heparin for preventing venous thromboembolism after lumbar decompressive surgery.

BACKGROUND: To evaluate the safety and effectiveness of argatroban for the prevention of venous thromboembolism (VTE) after posterior lumbar decompressive surgery. METHODS: Included in this retrospective study were 556 patients who underwent posterior lumbar decompressive surgery for trauma and degenerative diseases. They were divided into two groups: argatroban group (n = 274), and low molecular weight heparin (LMWH) group (n = 282). The occurrence of postoperative venous thrombosis and complications including hemorrhage and allergic reaction was compared between the two groups. Neurological and clinical outcomes in terms of Visual Analogue Scale (VAS) and the Oswestry Disability Index (ODI) were assessed before operation and at 6 and 12 months after operation. RESULTS: Postoperative venous thromboembolism (VTE) occurred in seven patient. No pulmonary embolism (PE) occurred in any patient. Thrombosis occurred in 3 cases (1.0%) and bleeding in 1 case (0.04%) in argatroban group vs. 4 (1.4%) and 4 (1.4%) in LMWH group, showing no significant between the two groups (P > 0.05). There was significant reduction in the severity of back and leg pain (VAS P < 0.05) and significant improvement in the patient quality of life (ODI, P < 0.05) 6 months and 1 year after operation, showing no significant difference between the two groups (P > 0.05). CONCLUSIONS: Argatroban proved to be equally effective as LWMH for anticoagulation therapy. Both drugs exhibited a similar preventive effect against postoperative VTE after posterior lumbar spine surgery, without increasing the risk of postoperative bleeding. The neurological and clinical outcomes are satisfactory and similar between the two pharmacological methods.

Coexistence of two ß-globin gene deletions in a Chinese girl with ß-thalassemia minor.

This study reports a rare case of ß(41/42,Cap)/ß(A) genotype in a girl with ß-thalassemia (ß-thal) minor. The 13-month-old Chinese proband suffered anemia, diarrhea, stunted growth and emaciation. The routine polymerase chain reaction-reverse dot-blot (PCR-RDB) test result for ß-thal mutations indicated that she was a compound heterozygote for ß(41/42) and ß(Cap). However, the complete blood cell (CBC) test gave the following results: mean corpuscular volume (MCV) 79.8 fL, mean corpuscular hemoglobin (MCH) 19.9 pg, with a Hb A2 value of 5.66%, suggesting that the proband also was ß-thal minor. The proband's father showed typical microcytic hypochromic anemia characteristics with a decreased MCV and MCH (63.1 fL and 20.9 pg, respectively) and an increased level of Hb A2 (5.60%), while the proband's mother had normal levels of MCV, MCH and Hb A2. The PCR-RDB test result showed her father was also a compound heterozygote for the ß(41/42) (HBB: c.126_129delCTTT) and ß(Cap) (HBB: c.-11_-8delAAAC) mutations and her mother was normal. Finally, DNA sequencing identified that the ß(41/42) and ß(Cap) mutations of the proband were inherited from her father and located on one ß-globin gene, suggesting that the proband's genotype is ß(41/42,Cap)/ß(A).

The relationship between cancer pain and quality of life in patients newly admitted to Wuhan Hospice Center of China.

To evaluate the relationship between pain and quality of life (QoL) in patients newly admitted to Wuhan Hospice Center, China. A total of 1,634 patients were analyzed in this retrospective study. A Numerical Rating Scale and Chinese-QoL instrument were used to assess pain score and QoL, respectively. Most patients experienced moderate to severe pain, which significantly impaired QoL. The pain was significantly correlated with appetite, mood, sleep, fatigue, pain intensity, daily activity, side effect, general appearance, and support from family. But there was no correlation with support from society, understanding of cancer, or attitude toward treatment. In our study, the relationship between pain and QoL was found to be reciprocal. The staff can offer a multidisciplinary care perspective for improving hospice care for this special group of population.

[Screening and identification of auto-antigen RHDAG1 of rheumatic heart disease].

OBJECTIVE: To identify the candidate auto-antigen of rheumatic heart disease as a molecular marker for this disease. METHODS: The total RNA of the heart tissue of patients with rheumatic heart disease was extracted and reverse-transcribed into long cDNA to construct the phage expression library. The library was screened using the serum from patients with active rheumatic fever, and the positive clone was identified and analyzed by bioinformatics and expressed in vitro. The expressed products were evaluated with Western blotting and its cross-reactivity was assessed. RESULTS: The phage expression library of the heart tissue of patients with rheumatic heart disease was constructed, with the titer of the primary library of 3.3×10(6) pfu/ml, recombinant rate of 99%, and 81% of the inserted segments were larger than 1 kb. An auto-antigen RHDAG1 was identified by screening, which was homologous to keratin 18. RHDAG1 was detected in the serum of patients with active rheumatic fever and of those with rheumatic heart disease, but not in the serum of healthy subjects. CONCLUSION: Phage display library can be an effective strategy to screen the auto-antigens of rheumatic heart disease. The auto-antigen RHDAG1 can be a candidate molecular biomarker of rheumatic heart disease and/or rheumatic fever.

Structural characterization and antioxidant activities of 2 water-soluble polysaccharide fractions purified from tea (Camellia sinensis) flower.

UNLABELLED: The water-soluble crude polysaccharide tea flower polysaccharide (TFP), obtained from tea (Camellia sinensis) flower by boiling-water extraction and ethanol precipitation, was fractionated by Sephadex G-100 column chromatography, giving 2 polysaccharide fractions termed TFP-1 and TFP-2. The structural features of TFP-1 and TFP-2 were investigated by high-performance liquid chromatography (HPLC), gel-permeation chromatography (GPC), rheometer, infrared (IR) spectra, nuclear magnetic resonance (NMR) spectroscopy, atomic force microscope (AFM), and scanning electron microscope (SEM). Results indicated that TFP-1 was composed of glucose: xylose: rhamnose: galactose=1.0:1.2:0.81:0.98 with a molecular weight of 167.5 KDa, while TFP-2 comprised glucose: xylose: rhamnose: arabinose=1.0:0.76:2.3:2.3 with a molecular weight of 10.1 KDa. The 1H NMR revealed that TFP-1 contained α-L-Rhap, α-D-Galp, α-D-GalpNAc, α-D-Xylp, α-D-Glcp, and ß-D-Glcp residues, while TFP-2 was illustrated to have α-L-Rhap, α-L-Arap, α-D-Xylp, α-D-Glcp, and α-D-GlcpNAc residues. Antioxidant activities of these fractions were investigated using various in vitro assay systems compared with ascorbic acid. In conclusion, TFP-2 exhibited the higher antioxidant activities and could be explored as a novel potential antioxidant. PRACTICAL APPLICATION: At present, commonly low-grade tea is preferred to extract the tea polysaccharide, to take full advantage of tea flower resource to extract polysaccharides can greatly reduce the cost of tea products. Thus, the search for plant-derived biomaterials from this study could generate natural value-added products from underutilized tea plant waste and used as a medicinal agent against chronic health problems, such as cancers, aging, and atherosclerosis caused by reactive free radicals that produced from oxidation.

Molecular characterization of a Chinese pedigree with beta-thalassemia intermedia.

Hereditary persistence of fetal hemoglobin (HPFH), often associated with mutations in the beta-globin gene cluster, is normally benign, but a person carrying both HPFH and another beta-thalassemia (beta-thal) mutation will develop serious anemia. These people might be erroneously diagnosed as having homozygous beta-thal with common reverse dot-blot methods. Here we report a 5-year old boy with thalassemia intermedia, who is a compound heterozygote for the rare HPFH-6 deletion with codons 41/42 (-TCTT) beta(0)-thal, who inherited the deletion from his mother and the beta(41/42) mutation from his father.

[Identification and tissue localization of intermediate filament protein in Angiostrongylus cantonensis].

OBJECTIVE: To identify the type of the intermediate filament (IF) protein of Angiostrongylus cantonensis and analyze its tissue localization. METHODS: Recombinant pET-IF of antigen IF was expressed in E.coli with IPTG induction, and the expression products were purified by His.Bind column and identified for determining the type of the IF protein by Western blotting. Anti-IF antibody was prepared by multi-spot subcutaneous injection into mouse and used to detect the tissue slices of A. cantonensis by immunohistochemical analysis. RESULTS: The antigen IF were correctly expressed and purified, and identified as a keratin located in the intestine wall and cytoplusma. CONCLUSION: The antigen IF is distributed in the intestine wall of A. cantonensis.

[Induced differentiation and signaling factor PTEN expression of 3T3-L1 adipocytes].

OBJECTIVE: To optimize the condition for inducing the differentiation of 3T3-L1 preadipocytes into adipocytes and study the expression of PTEN tumor suppression gene in this process, aiming to understand the regulatory role of PTEN in normal adipocyte differentiation and collect laboratory evidence for developing drugs targeting PTEN. METHODS: The differentiation of 3T3-L1 preadipocytes cultured in high-glucose DMEM were induced according to 2 protocols with different combinations of dexamethasone, isobutylmethylxanthine (IBMX) and insulin, and the resultant adipocytes were identified by oil red O staining. The total proteins of 3T3-L1 were extracted and analyzed by Western blotting, and PTEN homology between mice and human was analyzed by bioinformatic method. RESULTS: For optimized 3T3-L1 differentiation, 3T3-L1 cells were initially induced with the combination of 1 micromol/L dexamethasone, 0.5 mmol/L IBMX and 5 microg/ml insulin for 48 h, followed by treatment with 5 microg/ml insulin in 4.5 g/L glucose DMEM for 48 h, which resulted in high differentiation rate of 3T3-L1 cells (up to 90% on the 10th day) with unified morphology and size. PTEN expression varied quantitatively in the process of differentiation, especially low on the 12th day as compared with those measured on days 4, 6 and 9. The mice PTEN mRNA shared 96% homology and PTEN amino acid 100% homology with their human counterparts. CONCLUSION: Endogenous PTEN expression is down-regulated during 3T3-L1 differentiation, suggesting that PTEN may enhance insulin sensitivity and promote adipogenesis under physiological conditions.

[Green fluorescent protein as a tracer of bone marrow stromal cells in bone tissue engineering in rhesus].

OBJECTIVE: To observe the role of green fluorescent protein (GFP) in tracing rhesus bone marrow stromal cells (rBMSCs) during tissue-engineered bone formation in vivo. METHODS: Ad5.CMV-GFP was amplified by infecting QBI-293A cells, and the bone marrow was harvested from the ilium of adult male rhesus to obtain rBMSCs, which were cultured and passaged in vitro. GFP was transfected into the third-passage rBMSCs via adenovirus vector and the labeled cells were inoculated into absorbable HA scaffold and cultured for 3 days, with untransfected rBMSCs as control, before the cell-matrix compounds were implanted into the latissimus dorsi muscles of rhesus. Samples were harvested at 6 week and embedded in paraform, and ground sections of the bone tissue were prepared to observe green fluorescence under laser scanning confocal microscope. Propidium iodide staining of the sections was also performed for observation. RESULTS: The rBMSCs grew well after GFP transfection, and green fluorescence could be seen 24 h after the transfection and became stronger till 48 h, with a positive transfection rate beyond 80%. Six weeks after cell implantation, the rBMSCs labeled by GFP-emitted green fluorescence were detected in the bone tissue under laser scanning confocal microscope. CONCLUSION: GFP can effectively trace BMSCs during bone tissue engineering, and the transplanted BMSCs constitute the main source of bone-forming cells in bone tissue engineering.