Pharmacokinetic study of Strongylocentrotus nudus egg polysaccharide in rats and beagles using a 3H-labeling method.

Strongylocentrotus nudus egg polysaccharide (SEP) extracted from sea urchins has potential anticancer activity. However, little is known about its pharmacokinetic properties. To investigate the pharmacokinetics of SEP, it was radiolabeled with tritium. Furthermore, a sensitive, selective, and rapid liquid scintillation counter (LSC) method for quantifying 3H-SEP in biological matrix was validated. The lower quantification limit of the method was 4 Bq. The relative standard deviations (RSDs) of the intra- and inter-day precision were <3.0% and <3.9%, respectively. 3H-SEP was successfully applied to investigate the pharmacokinetics of SEP after intravenous administration of 20, 40, and 80 mg/kg (40 µCi/kg) in rats and 5, 10, and 20 mg/kg (6 µCi/kg) in beagles. The AUC(0-t) of SEP at three different doses was 487.81 ± 39.99 mg/L*h, 1,003.10 ± 95.94 mg/L*h, and 2,188.84 ± 137.73 mg/L*h in rats and 144.12 ± 3.78 mg/L*h, 322.62 ± 28.03 mg/L*h, and 754.17 ± 37.79 mg/L*h in beagles. The terminal elimination half-life (t1/2) of SEP was longer in beagles (204.29 ± 139.34 h) than in rats (35.48 ± 6.04 h). The concentration of SEP in plasma declined rapidly in both rats and beagles. All the study results provide detailed pharmacokinetic profiles of SEP in two kinds of animals, which will be helpful for further development.

Effect of Surgical Clipping versus Endovascular Coiling on the Incidence of Delayed Cerebral Ischemia in Patients with Aneurysmal Subarachnoid Hemorrhage: A Multicenter Observational Cohort Study with Propensity Score Matching.

OBJECTIVE: The effect of surgical clipping (SC) and endovascular coiling (EC) on the incidence of delayed cerebral ischemia (DCI) in patients with aneurysmal subarachnoid hemorrhage (aSAH) has always been a controversial topic. Hence, it is necessary to reanalyze the effects of the 2 surgical methods on DCI, which determines the choice of the most favorable method for patients who are suitable for both surgical modalities. METHODS: A multicenter retrospective observational cohort study was performed to evaluate all consecutive patients with aSAH admitted to 5 medical centers in China between April 2019 and June 2021. Univariable and multivariable analyses were used to confirm risk factors of DCI after aSAH. A 1:1 propensity score matching model was generated in the EC and SC groups to reduce the influence of all confounding factors on DCI. RESULTS: A total of 412 patients were included, and 115 patients (27.9%) developed DCI. After propensity score matching for controlling demographic information, past medical history, admission clinical status, aneurysm characteristics, and inflammatory factors associated with DCI, 133 patients with SC and 133 patients with EC treatment were matched. The results of the matched cohorts indicate a significantly lower incidence of DCI when patients received EC than SC (31.9% vs. 20%; adjusted odds ratio, 1.87; 95% confidence interval, 1.08-3.29; P = 0.027). CONCLUSIONS: The study found that the patients who received SC treatment had a higher incidence of DCI than did those who received EC and suggested that ruptured intracerebral aneurysm is preferentially coiled rather than clipped if the aneurysm is suitable for both surgical modalities.

Development and validation of a liquid chromatography-tandem mass spectrometry method for CPUL1, a novel antitumor candidate compound, and its application to pharmacokinetic studies.

An active substance of pyrano[3,2-a]phenazine, also called CPUL1, is a synthesized phenazine derivative and displays broad-spectrum anticancer activities. Quantitative assessment of CPUL1 in biological samples has not been well established, hindering pharmaceutical development and application. According to international guidelines, a sensitive and selective liquid chromatography-tandem mass spectrometry method in negative ion mode was developed and validated for quantification of CPUL1 in human plasma, colorectal cancer cell lines, and rat plasma, whereby linearity and accuracy were demonstrated for the range of 1-1000 ng/ml. The validated liquid chromatography-tandem mass spectrometry method was successfully employed in pharmacokinetic studies of CPUL1 in vitro and in vivo. Notably, the cellular pharmacokinetic behavior of CPUL1 varies in colorectal cancer cell lines. Regarding the pharmacokinetic processes in vivo, oral absorption was less effective than an injection, with a bioavailability of 23.66%. CPUL1 was linearly eliminated after a single administration; however, it could accumulate in tissues (heart, liver, spleen, lung, and kidney) after multiple injections. In summary, this study established a capable bioanalytical method for CPUL1 and provided exploratory pharmacokinetic data, paving the way for use of this promising derivative in disease models.

Kiaa0101 serves as a prognostic marker and promotes invasion by regulating p38/snail1 pathway in glioma.

BACKGROUND: Kiaa0101, a regulator of cell proliferation, is overexpressed in many malignant tumors. However, its role in promoting invasion of glioma is poorly understood. Here, we investigated the effects of Kiaa0101 on glioma invasion and elucidated the underlying mechanisms of action. METHODS: We analyzed Kiaa0101 expression using datasets from four public databases, namely TCGA, CGGA, Gravendeel and Rembrandt as well as experimentally on 123 glioma samples via western blot (WB), RT-PCR and immunohistochemistry (IHC). We further quantified migration and invasion using wound healing and transwell assays. WB, IHC and immunofluorescence (IF) were used to detect expression of invasion related markers. Moreover, we detected tumor invasion of glioma cells in vivo in 5-week-old Balb/c nude mice. RESULTS: Kiaa0101 was upregulated in glioma, relative to non-tumor brain tissues, with the expression increasing with increase in glioma grade. Kiaa0101 mRNA expression was especially enriched in isocitrate dehydrogenase (IDH)1 wild-type glioma. Kaplan-Meier analysis, based on the aforementioned datasets, revealed that high Kiaa0101 levels were significantly associated with worse overall survival. Besides, shRNA-mediated Kiaa0101 knockdown inhibited migration and invasion of glioma cells by reducing snail1 expression both in vitro and in vivo, whereas its upregulation enhanced malignant behaviors of these cells. Furthermore, Kiaa0101 regulated snail1 expression by activating the p38MAPK signaling pathway. CONCLUSIONS: Our findings strongly indicate that Kiaa0101 is a prognostic biomarker for malignant tumors, and its inhibition may be an effective strategy for treating glioma.

Antibacterial constituents from Antarctic fungus, Aspergillus sydowii SP-1.

One new alkaloid, named as acremolin C (1), was isolated from static culture of Antarctic fungus, Aspergillus sydowii SP-1, in an investigation of the antimicrobial constituents of this Antarctic microorganism, and its structure was determined by spectroscopic methods. Additionally, four known compounds, cyclo-(L-Trp-L-Phe) (2), 4-hydroxyphenylacetic acid (3), (7S)-(+)-hydroxysydonic acid (4) and (7S, 11S)-(+)-12-hydroxysydonic acid (5), were isolated and identified. Biological studies disclosed that compounds 2, 4 and 5 showed moderate inhibitions against methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus epidermidis (MRSE) as comparing to tigecycline, while compound 1 displayed weak inhibition activities against MRSA and MRSE.

Design, combinatorial synthesis and biological evaluations of novel 3-amino-1'-((1-aryl-1H-1,2,3-triazol-5-yl)methyl)-2'-oxospiro[benzo[a] pyrano[2,3-c]phenazine-1,3'-indoline]-2-carbonitrile antitumor hybrid molecules.

A combinatorial chemical library of fifty-nine novel 3-amino-1'-((1-aryl-1H-1,2,3-triazol-5-yl)methyl)-2'-oxospiro[benzo[a]pyrano[2,3-c]phenazine-1,3'-indoline]-2-carbonitrile, designed as hybrid molecules of phenazine, pyran, indole and 1,2,3-triazole pharmacophores, were constructed in this study. Cytotoxic evaluation indicated that some compounds exhibited moderate cytotoxicity against HCT116, MCF7, HepG2 and A549 cancer cell lines in vitro, in which compound 36 was found to have best antiproliferative activity against the A549 cancer cell line with IC50 value of 5.4 µM. All compounds had low or no effect against L02 and HUVEC non-cancer cell lines. Compound 36 was further confirmed to mainly locate mitochondria in A549 cancer cells via laser-scanning confocal microscopy. Moreover, compound 36 was proved to increase ROS production and induce cell cycle arrest in S phase. Western blot analysis illustrated Bax/Bcl-2 ratio was increased at dose-dependent manner, and both cleaved caspase-3 and cleaved caspase-9 was enhanced by treated with compound 36. All the above evidences in vitro indicated that compound 36 might induce the apoptosis of A549 cancer cells via a mitochondria-dependent pathway.

[Absorption, distribution and elimination of (59)Fe- corn polysaccharide iron complex in rats: a study with radioactivity isotope tracing].

OBJECTIVE: To establish a method for detecting plasma concentration of corn polysaccharide iron complex (CPIC) and investigate its absorption, distribution and elimination in rats. METHODS: Using radioactivity isotope tracing method, we detected the radioactivity of (59)Fe-CPIC in the plasma of rats at different time points by gavages of 3 doses (28.0, 14.0, and 7.0 mg/kg) (59)Fe-CPIC in SD rats. The pharmacokinetic parameters was obtained using DAS 2.0 program for analysis of tissue distribution and elimination of (59)Fe-CPIC. RESULTS: The standard curve was linear within the range of 0.14-141 µg/ml (r=0.9999, n=5). The average recovery was 95% with a relative standard deviation no more than 15%. The pharmacokinetic parameters at 3 doses obtained, namely t1/2 and AUC (0-), were 214∓104, 231∓110, and 181∓81 min, and 1986.3∓513.3, 737.0∓467.0, and 315.1∓226.1 mg·min-1·L(-)1, respectively. (59)Fe-CPIC were detected in all the 13 tissues types examined and high radioactivity intensity was found in the gastrointestinal tract, hematogenic organs and other organs rich in blood. (59)Fe-CPIC was eliminated after intragastric administration primarily via the feces in rats. CONCLUSION: The method we established is easy and specific, and the pharmacokinetic parameters of (59)Fe-CPIC fit the two- compartment open model.

Prediction and identification of potential immunodominant epitopes in glycoproteins B, C, E, G, and I of herpes simplex virus type 2.

Twenty B candidate epitopes of glycoproteins B (gB2), C (gC2), E (gE2), G (gG2), and I (gI2) of herpes simplex virus type 2 (HSV-2) were predicted using DNAstar, Biosun, and Antheprot methods combined with the polynomial method. Subsequently, the biological functions of the peptides were tested via experiments in vitro. Among the 20 epitope peptides, 17 could react with the antisera to the corresponding parent proteins in the EIA tests. In particular, five peptides, namely, gB2(466-473) (EQDRKPRN), gC2(216-223) (GRTDRPSA), gE2(483-491) (DPPERPDSP), gG2(572-579) (EPPDDDDS), and gI2(286-295) (CRRRYRRPRG) had strong reaction with the antisera. All conjugates of the five peptides with the carrier protein BSA could stimulate mice into producing antibodies. The antisera to these peptides reacted strongly with the corresponding parent glycoproteins during the Western Blot tests, and the peptides reacted strongly with the antibodies against the parent glycoproteins during the EIA tests. The antisera against the five peptides could neutralize HSV-2 infection in vitro, which has not been reported until now. These results suggest that the immunodominant epitopes screened using software algorithms may be used for virus diagnosis and vaccine design against HSV-2.

[The relationship between coagulation/anticoagulation imbalance and oxidative stress in patients with chronic obstructive pulmonary disease].

OBJECTIVE: To explore the relationship between coagulation/anticoagulation imbalance and oxidative stress in the patients with chronic obstructive pulmonary disease during acute exacerbation (AECOPD) before and after treatment. METHODS: Plasma tissue factor (TF) and tissue factor pathway inhibitor (TFPI) activity was detected by chromogenic assay in 28 AECOPD patients before and after treatment as well as in 30 healthy controls. The total antioxidative capacity (TAC), malondialdehyde (MDA) and glutathione peroxidase (GSH-PX) in plasma were measured in both groups. RESULTS: The levels of plasma TF and TFPI, and their ratio (TF/TFPI) in AECOPD patients before treatment were significantly higher than those after treatment (all P < 0.01), the latter were still higher than those in the healthy persons (all P < 0.01). The levels of the TAC and GSH-PX in plasma in AECOPD patients before treatment were significantly lower than those after treatment (all P < 0.01), the latter were still lower than those in the healthy persons (all P < 0.01). The plasma MDA in AECOPD patients before treatment was significantly higher than that after treatment (P < 0.01), which was still higher than that in the healthy persons (P < 0.05). There were negative correlations between TF/TFPI ratio and TAC (r = -0.518, P < 0.01), GSH-PX (r = -0.454, P < 0.05), PaO2 (r = -0.511, P < 0.01) respectively and a positive correlation between TF/TFPI ratio and the percentage of neutrophils (r = 0.379, P < 0.05) in AECOPD patients before treatment. There still were negative correlations between TF/TFPI ratio and TAC (r = -0.420, P < 0.05), FEV(1)% to predicted (r = -0.480, P < 0.05) respectively, and a positive correlation between TF/TFPI ratio and MDA (r = 0.451, P < 0.05) in AECOPD patients after treatment. CONCLUSIONS: There existed coagulation/anticoagulation imbalance and oxidation/antioxidation imbalance before and after treatment in AECOPD patients and their relationship was explored.

[Effects of rosiglitazone on peroxisome proliferator activated receptor-γ pathway in patients with chronic obstructive pulmonary disease].

OBJECTIVE: To explore the effects of rosiglitazone on peroxisome proliferator activated receptor-γ (PPAR-γ), nuclear factor-κB and tumor necrosis factor-α (TNF-α) in patients with chronic obstructive pulmonary disease (COPD). METHODS: From Apr. 2010 to Nov. 2010, 30 patients with acute exacerbations of COPD, 22 males and 8 females, age 54 - 87 (mean 72 ± 9) years and 24 healthy controls, 18 males and 6 females, age 52 - 80 (mean 69 ± 10) years were included. The peripheral blood mononuclear cells (PBMCs) were isolated from venous blood and then cultured. On the basis of the treatment given, the PBMCs of COPD patients were divided into 3 groups: non-treatment group, rosiglitazone treatment group (rosiglitazone group) and rosiglitazone and GW9662 treatment group (combined treatment group). Cells from the healthy controls (control group) did not receive any drug treatment. The mRNA expression of PPAR-γ and NF-κB was measured with real-time PCR. The protein expression and nuclear translocation of PPAR-γ and NF-κB were detected using immunofluorescence with laser scanning confocal microscopy. The TNF-α level in culture supernatant was measured with ELISA. One-way ANOVA and LSD-t test and the Pearson correlation coefficient were used for statistical analysis. RESULTS: The mRNA and protein levels of PPAR-γ were lower in the non-treatment group (0.52 ± 0.10, 55 ± 11) than those in the control group (1, 85 ± 9), while the levels of NF-κB mRNA and protein were higher in the non-treatment group (1.69 ± 0.07, 145 ± 17) than those in the control group (1, 118 ± 7). The mRNA and protein levels of PPAR-γ in the rosiglitazone group (4.47 ± 0.11, 204 ± 12) were significantly increased compared with the non-treatment group, while the mRNA and protein levels of NF-κB (0.33 ± 0.04, 59 ± 14) were remarkably decreased compared with the non-treatment group. The mRNA and protein levels of PPAR-γ (2.25 ± 0.31, 142 ± 23) were significantly decreased in the combined treatment group compared to the rosiglitazone group, but higher compared with the non-treatment group, while the mRNA and protein levels of NF-κB (0.64 ± 0.02, 90 ± 10) were increased compared with the rosiglitazone group, but decreased compared to the non-treatment group (F = 29.21 - 567.42, all P < 0.01). The TNF-α level was significantly higher in the non-treatment group (96.2 ± 1.4) µg/L than that in the control group (85.3 ± 1.0) µg/L. The TNF-α level in the rosiglitazone group (63.0 ± 2.5) µg/L was remarkably decreased compared with the non-treatment group, while that in the combined treatment group (83.3 ± 1.9) µg/L was increased compared with the rosiglitazone group, but decreased compared to the non-treatment group (F = 293.72, P < 0.01). The proteins of PPAR-γ and NF-κB were respectively located in cytoplasm and in nucleus in the non-treatment group, meanwhile they were located in both cytoplasm and nucleus in the control group. PPAR-γ protein was translocated from cytoplasm into nucleus and NF-κB protein was translocated from nucleus into cytoplasm in the rosiglitazone group. In the combined treatment group, PPAR-γ protein translocated from nucleus into cytoplasm and NF-κB protein partly translocated from cytoplasm into nucleus. By linear correlation analysis, PPAR-γ protein was negatively correlated with NF-κB protein and TNF-α level (r = -0.935, -0.924, all P < 0.01), while NF-κB protein was positively correlated to TNF-α level (r = 0.846, P < 0.01). CONCLUSIONS: The expression and activity of PPAR-γ were decreased in COPD patients. PPAR-γ agonist rosiglitazone inhibited inflammation in COPD through upregulating the expression and activity of PPAR-γ and inhibition of NF-κB and TNF-α. It suggests that PPAR-γ may play an important role in the inflammation of COPD.

Establishment of a novel model for studying the effects of extracts of Chinese herb medicine on human type II 5alpha-reductase in vitro.

Human steroid 5alpha-reductase type II (hSRD5A2) and dihydrotestosterone (DHT) play important roles in benign prostatic hyperplasia (BPH). The aim of our study was to establish a novel model to investigate the inhibitory effects of extracts and compounds of Chinese herb medicine on hSRD5A2. The gene, hSRD5A2, was artificially synthesized and cloned into pcDNA3.1(+) vector, which was transfected into CHO cells by liposome. Transfected cells were screened through G418 and MTX. The expressed protein of hSRD5A2 by cells was purified and detected by western blotting. A minimum reactive system comprising hSRD5A2 and testosterone (T) as substrate together with NADPH as hydrogen donor was established for screening inhibitors of hSRD5A2. The reaction system was optimized in the concentrations of T, NADPH, and hSRD5A2 and reaction temperature, time, and activity of hSRD5A2 were determined by the production of DHT. Furthermore, we screened some extracts and compounds of Chinese herb medicine using this model. The concentrations of T, NADPH, and hSRD5A2 were 0.02 microM, 0.8 mM, and 0.05 U/microl, respectively, in the model; maximum activity of hSRD5A2 was achieved at 37 degrees C and 60 min reaction, and mangiferin had significant inhibitory effect on the activity of hSRD5A2. The model in this study is convenient and reliable for screening and evaluation of inhibitors of hSRD5A2; mangiferin may be a potential medicine for the treatment of BPH.

Recombinant expression of rt-PA gene (encoding Reteplase) in gametophytes of the seaweed Laminaria japonica (Laminariales, Phaeophyta).

The life cycle of seaweed Laminaria japonica involves a generation alternation between diploid sporophyte and haploid gametophte. The expression of foreign genes in sporophte has been proved. In this research, the recombinant expression in gametophyte was investigated by particle bombardment with the rt-PA gene encoding the recombinant human tissue-type plasminogen activator (Reteplase), which is a thrombolytic agent for acute myocardial infarction (AMI). Transgenic gametophytes were selected by their resistance to herbicide phosphiothricin (PPT), and proliferated in an established bubble column photo-bioreactor. According to the results from quantitative ELISA, Southern blotting, and fibrin agarose plate assay (FAPA) for bioactivity, it was showed that the rt-PA gene had been integrated into the genome of gametophytes of L. japonica, and the expression product showed the expected bioactivity, implying the proper post-transcript modification in haploid gametophyte.