pLARmEB: integration of least angle regression with empirical Bayes for multilocus genome-wide association studies.

Multilocus genome-wide association studies (GWAS) have become the state-of-the-art procedure to identify quantitative trait nucleotides (QTNs) associated with complex traits. However, implementation of multilocus model in GWAS is still difficult. In this study, we integrated least angle regression with empirical Bayes to perform multilocus GWAS under polygenic background control. We used an algorithm of model transformation that whitened the covariance matrix of the polygenic matrix K and environmental noise. Markers on one chromosome were included simultaneously in a multilocus model and least angle regression was used to select the most potentially associated single-nucleotide polymorphisms (SNPs), whereas the markers on the other chromosomes were used to calculate kinship matrix as polygenic background control. The selected SNPs in multilocus model were further detected for their association with the trait by empirical Bayes and likelihood ratio test. We herein refer to this method as the pLARmEB (polygenic-background-control-based least angle regression plus empirical Bayes). Results from simulation studies showed that pLARmEB was more powerful in QTN detection and more accurate in QTN effect estimation, had less false positive rate and required less computing time than Bayesian hierarchical generalized linear model, efficient mixed model association (EMMA) and least angle regression plus empirical Bayes. pLARmEB, multilocus random-SNP-effect mixed linear model and fast multilocus random-SNP-effect EMMA methods had almost equal power of QTN detection in simulation experiments. However, only pLARmEB identified 48 previously reported genes for 7 flowering time-related traits in Arabidopsis thaliana.

A case report of fetal malignant immature mediastinal teratoma.

PURPOSE: Fetal immature mediastinal teratoma is a rare disease. The pressure generated by the tumor mass can cause hydrops fetalis, pulmonary hypoplasia, pleural and peritoneal effusion, and polyhydramnios which cause the death of the fetus. Routine prenatal ultrasound has enabled accurate diagnosis. MATERIALS AND METHODS: The authors report a 26-year-old patient, gravida 4 para 1, who was referred to this hospital, carrying a fetus with immature mediastinal teratoma. RESULTS: At 27 weeks of gestation, a routine prenatal ultrasound suggested the fetus had a mass at the anterior mediastinum, accompanied by pulmonary hypoplasia, pleural and peritoneal effusion, subcutaneous edema of head and chest, and polyhydramnios. After the therapeutic abortion, the gross anatomy confirmed the mediastinal mass. The histological examination showed that the mass was a grade 2 immature teratoma. CONCLUSIONS: The mother of the fetus had been exposed to plaster, paint, and paint-thinner in the first trimester of pregnancy, suggesting that these chemical contacts may be one of the causes of the disorder.

[Epidemiological characteristics of mpox epidemic in Guangzhou].

Objective: To understand the epidemiological characteristics of mpox epidemic in Guangzhou and provide scientific evidence for the prevention and control of the disease. Methods: Based on the mpox surveillance system in Guangzhou, suspected mpox cases with fever and rash were reported by local hospitals at all levels to centers for disease control and prevention in Guangzhou for sampling, investigation and diagnosis. Descriptive epidemiological analysis was conducted on the clinical characteristics and treatment of the mpox cases and positive detection rate reported in Guangzhou as of 24:00 on June 23. Whole genome sequencing of the virus isolates was performed using Illumina Miniseq high-throughput sequencing platform. Results: The first mpox case in Guangzhou was reported on June 10 in 2023. As of 24:00 on June 23, a total of 25 confirmed mpox cases were reported. All the mpox cases were men with a M(Q1,Q3) of 32 (26, 36) years, the majority of the cases were MSM (96.0%). The main clinical features were rash (100.0%, 25/25), lymphadenectasis (100.0%, 25/25) and fever (52.0%, 13/25). Rash usually occurred near the genitals (88.0%, 22/25). The close contacts, mainly family members (40.4%, 23/57), showed no similar symptoms, such as fever or rash. The positive rate of mpox virus in household environment samples was 30.5%. The analyses on 3 complete gene sequences of mpox virus indicated that the strains belonged to West African type Ⅱb clade, B.1.3 lineage. Conclusions: Hidden transmission of mpox virus had occurred in MSM in Guangzhou. However, the size of affected population is relatively limited, and the possibility of wide spread of the virus is low.

Activation of imidazoline I-2B receptors by allantoin to increase glucose uptake into C2C12 cells.

Allantoin, an active principle of the yam, belongs to the group of guanidinium derivatives and has been reported to lower plasma glucose in diabetic animals. Recent evidence indicates that activation of the imidazoline I(2B) receptor (I(2B)R) by guanidinium derivatives also increases glucose uptake; however, the effect of allantoin on I(2B)R is still unknown. Glucose uptake into cultured C2C12 cells was determined using 2-[¹4C]-deoxy-D-glucose as a tracer. The changes in 5'-AMP-activated protein kinase (AMPK) expression were also identified by Western blotting analysis. The allantoin-induced glucose uptake action was dose-dependently blocked by BU224, a specific I2R antagonist, in C2C12 cells. Moreover, AMPK phosphorylation by allantoin was found to be dose-dependently increased in C2C12 cells using AICAR treatment as a reference. In addition, both actions of allantoin, the increases in glucose uptake and AMPK phosphorylation, were dose-dependently attenuated by amiloride in C2C12 cells. Moreover, compound C at concentrations sufficient to inhibit AMPK blocked the allantoin-induced glucose uptake and AMPK phosphorylation. Thus, we suggest that allantoin can activate I(2B)R to increase glucose uptake into cells, and propose I(2B)R as a new target for diabetic therapy.

Plasma glucose-lowering action of allantoin is induced by activation of imidazoline I-2 receptors in streptozotocin-induced diabetic rats.

Allantoin, an active principle of yam, is documented to lower plasma glucose in diabetic rats. However, action mechanisms of allantoin remain obscure. It has been indicated that metformin shows ability to activate imidazoline I-2 receptors (I-2R) to lower blood sugar. Allantoin has also a chemical structure similar to metformin; both belong to guanidinium derivative. Thus, it is of special interest to know the effect of allantoin on I-2R. In the present study, the marked plasma glucose-lowering action of allantoin in streptozotocin-induced type-1 like diabetic rats was blocked by specific I-2R antagonist, BU224, in a dose-dependent manner. Also, the increase of ß-endorphin release by allantoin was blocked by BU224 in the same manner. Otherwise, amiloride at the dose sufficient to block I-2AR abolished the allantoin-induced ß-endorphin release and inhibited the blood glucose-lowering action of allantoin markedly but not completely. The direct effect of allantoin on glucose uptake in isolated skeletal muscle was also blocked by BU224. Also, the phosphorylation of AMPK in isolated skeletal muscle was raised by allantoin in a concentration-dependent manner. More-over, insulin sensitivity in diabetic rats was markedly increased by allantoin and this action was also blocked by BU224. These results suggest that allantoin has an ability to activate imidazoline I-2R while I-2AR is linked to the increase of ß-endorphin release and I-2BR is related to other actions including the influence in skeletal muscle for lowering of blood glucose in type-1 like diabetic rats. Thus, allantoin can be developed to treat diabetic disorders in the future.

Efficient inhibition of cisplatin-resistant human ovarian cancer growth and prolonged survival by gene transferred vesicular stomatitis virus matrix protein in nude mice.

BACKGROUND: The vesicular stomatitis virus matrix protein (VSVMP) has been receiving attention as an anticancer agent because of its ability of inducing apoptosis. MATERIALS AND METHODS: Nude mice bearing A2780s and A2780cp ovarian tumors were treated twice weekly with i.v. administration of 50 microg VSVMP/250 mug liposome complex, 50 microg empty plasmid/250 microg liposome complex, 0.9% NaCl solution or weekly with i.p. administration of cisplatin (5 mg/kg) for 3 weeks. Tumor volume and survival time were observed. TUNEL assay and CD34 vessel staining were conducted in tumor tissue. Antiangiogenesis in vivo were determined by sponge assay. Antiproliferative and apoptosis-inducing activities of VSVMP in vitro were tested on MS1 murine endothelial cells and four human ovarian cancer cell lines: A2780s, A2780cp, HO8910 and COC1. RESULTS: Administration of VSVMP resulted in significant inhibition (87%-98% maximum inhibition relative to controls) in the growth of A2780s and A2780cp tumor xenografts, and prolonged the survival of the treated mice. Complete tumor regression happened in VSVMP-treated mice in both tumor models. These antitumor responses were associated with marked increases in tumor apoptosis and reductions in intratumoral microvessel density. CONCLUSIONS: Our data indicate that VSVMP may provide an effective approach to inhibit both cisplatin-sensitive and -resistant human ovarian cancer growth with minimal side-effects.

Development of an indirect enzyme-linked immunosorbent assay (ELISA) to differentiate antibodies against wild-type porcine reproductive and respiratory syndrome from the vaccine strain TJM-F92 based on a recombinant Nsp2 protein.

An accurate ELISA method to differentiate pigs infected with wild-type porcine reproductive and respiratory syndrome (PRRSV) strains from vaccinated ones would help to monitor PRRSV vaccination compliance. The recombinant protein GST-d120aa derived from the continuous deletion of 120 amino acids in the non-structural protein 2 region of the modified-live vaccine strain TJM-F92 was used to develop an indirect enzyme-linked immunosorbent assay (d120-ELISA) for differentiating serum antibodies against TJM-F92 from other PRRSV strains. At the optimized cut-off value which was calculated at an S/P of 0.25, it yielded a sensitivity of 90.7% and a specificity of 95.1%. Cross-reactivity tests suggested that the d120-ELISA was PRRSV-specific. Coefficient of variations of the repeatability tests ranged between 1.41-17.02%. The results suggest that the d120-ELISA is suitable for differentiating animals infected with wild-type strains from those immunized with MLV TJM-F92.

Bioaugmented soil aquifer treatment for P-nitrophenol removal in wastewater unique for cold regions.

P-nitrophenol (PNP) is a toxic and recalcitrant organic pollutant and a usual intermediate in the production of fine chemicals, which has posed a significant threat to subsurface environment safety. Soil aquifer treatment (SAT) is a promising method to remove and remediate contamination in vadose zone with low cost and high efficiency. However, there are still research gaps for the treatment of recalcitrant contaminants by SAT in cold regions, such as un-robust indigenous microbes and low temperature constraint in vadose zone. The bioaugmentation technology was first introduced into SAT in order to enhance the removal ability of PNP by SAT operated in cold regions in this study. A high-efficiency PNP-degrading bacterium was successfully isolated, which can efficiently degrade PNP below 200 mg L-1 with a degradation rate above 99% at 15 °C close to the real subsurface temperature in cold regions, and added into SAT for bioaugmentation. The feasibility of bioaugmented SAT and associated PNP removal process were investigated by laboratory sand columns, along with effects of the SAT operative parameters (namely PNP loading concentration, flow rate and soil saturation level of SAT). Within the range of PNP loading stresses tested (1-200 mg L-1), PNP removal efficiency was optimal at constant flow rate of 219 mL d-1 in unsaturated operating condition of SAT under 15 °C among all the investigated experimental conditions. Longer hydraulic residence time increased the PNP removal rate, although the accumulated mass removed reduced and the removal efficiencies remained constant in unsaturated operating condition of SAT. It is found from the comparison between the PNP removals via both unsaturated and saturated columns that slight difference only in the removal rate of PNP was observed and the highly efficient bioaugmented SAT can completely degrade PNP of 10 mg L-1 within 5 wetting/drying cycles under both scenarios.

Humoral and cellular immunity induced by tumor cell vaccine based on the chicken xenogeneic homologous matrix metalloproteinase-2.

Matrix metalloproteinase-2 (MMP-2) has been used as a target for cancer immunotherapy. The activation of immunization by breaking immune tolerance to self-MMP-2 may be one of the promising approaches for the treatment of MMP-2-positive tumors. In this study, we constructed the xenogeneic tumor cell vaccine c-MMP-2 by transfecting CT26 and LLC cells with chicken MMP-2 cDNA constructs. MMP-2-specific autoantibodies in sera and tumor cells were found in mice immunized with c-MMP-2. Protection against tumor growth was evaluated in respect of the relative contributions of autoantibodies, CD4+, and CD8+ T cells. Treatment with this vaccine (c-MMP-2) also prolonged the survival time of mice bearing cancer. The specific cytotoxic T-cell responses suggested that the treatment increased CD8+ T-cell activity. The antitumor activity of c-MMP-2 was abrogated by in vivo depletion of CD4+ and CD8+ T-lymphocytes and improved by adoptive transfer of CD4+ and CD8+ T-lymphocytes from the mice treated with c-MMP-2. An alternative DNA vaccination strategy for cancer therapy was identified in this study by eliciting humoral and cellular immunoresponse with a crossreacting transfectant.

Increasing upstream capacity in TDM-PON with multiple-wavelength transmission using Fabry-Perot laser diodes.

We propose a new technique for multiple-wavelength upstream transmission in time division multiplexed-passive optical networks using Fabry-Perot laser diodes (FP-LD) at optical network units (ONU). The FP-LD transmits at one of strategically separated seeding wavelengths from the optical line terminal enabling the ONUs to join one of few TDM upstream channels. The scheme increases upstream capacity without the use of costly, higher speed burst mode transceivers. We present experimental results showing that up to 9 upstream channels at 2.5 Gb/s data rate can be achieved with this scheme. The paper presents locking characteristics of the FP-LD relevant for this application such as suppression of other seeding wavelengths, minimum wavelength separation and burst mode operation.

Dynamic progression of an intraperitoneal xenograft model of human ovarian cancer and its potential for preclinical trials.

This study was conducted to establish a feasible intraperitoneal xenograft model in nude mice which mimicked the dynamic progression of human ovarian cancer and to explore its potential for preclinical trials. A human ovarian tumor line SKOV3 was originally injected s.c. to develop tumors; then the tumors were harvested and minced into small particles for i.p. inoculation in three groups of nude mice which would be monitored consecutively. The intraperitoneal carcinomatosis and relevant organs were collected for histopathological dynamic comparison and CA125 immunohistochemical staining 7, 21 and 49 days after inoculation. An additional experiment with cisplatin sensitivity test was performed and tumor tissues were observed for apoptosis-Hoechst assay. The intraperitoneal carcinomatosis had a rapid progression which resulted in extensive dissemination on the peritoneal surfaces and invasion into abdominal lymph nodes, livers, pancreas and spleen. Tumor tissues revealed similar morphological features of primary tumor from which SKOV3 derived and part of in vivo tumor mass was positive for CA125. Cisplatin could significantly inhibit the intraperitoneal carcinomatosis growth. This model may provide a valuable platform to study the biological properties of ovarian cancer as well as to test new therapeutic strategies in preclinical trials.

Systemic inhibition of tumor growth by soluble Flk-1 gene therapy combined with cisplatin.

Soluble Flk-1, a soluble vascular endothelial growth factor (VEGF) receptor, is a potent inhibitor of angiogenesis, which could restrain growth and metastasis of some experimental tumors. However, antiangiogenic agents alone cannot eradicate tumor completely, and should be combined with other therapy to enhance their effects. In this study, we evaluated the antitumor activity of the combination therapy in the immunocompetent BALB/c mice bearing H22 hepatoma and Meth A fibrosarcoma, respectively. Mice were treated with either msFlk-1 i.m. at 100 microg/mouse once every 3 days for four times from day 3 after the tumor cell injection, cisplatin cycled twice (2 mg/kg i.p. on days 4 and 11 after the tumor cell inoculation), or both agents together. Tumor growth and survival time were continually observed. Antiangiogenesis in vivo was determined by CD31 immunohistochemistry. Assessment of apoptotic cells and histological analysis was also conducted in tumor tissues. Our results showed that the combination therapy could evidently improve antitumor efficacy, including tumor growth suppression, mice survival prolongation, tumor cell apoptosis augmentation as well as neovascularization inhibition as compared with controls, without serious adverse effects. Our data suggest that the combination of DDP with msFlk-1 is more effective to suppress tumor growth in mice than either agent alone, and this combination regimen showed its potential for future clinical application.

Idiotypic protein-pulsed adherent peripheral blood mononuclear cell-derived dendritic cells prime immune system in multiple myeloma.

Adherent peripheral blood mononuclear cell-derived dendritic cells pulsed with autologous idiotypic protein (Id) were given to a patient with advanced-stage refractory myeloma. Potentially beneficial antimyeloma Id-specific immune responses were produced, characterized by MHC-dependent T-cell-proliferative responses with cytokine release and the production of anti-Id antibodies. A T-cell line generated after vaccination was also able to lyse autologous Id-pulsed targets and recognize fresh autologous myeloma cells. The immune responses were associated with a transient minor fall in the serum Id level and were not ablated by high-dose myeloablative chemotherapy. This report therefore demonstrates the clinical use of adherent peripheral blood mononuclear cell-derived dendritic cells for vaccination in cancer and the persistence of immune responses after high-dose chemotherapy. Such a therapeutic approach may be useful in reducing the relapse rate in patients who have minimal residual disease after chemotherapy.

New aristolochic acid, aristololactam and renal cytotoxic constituents from the stem and leaves of Aristolochia contorta.

Two novel phenanthrene derivatives, aristololactam IVa (1) and 9-hydroxy aristolochic acid I (2) were isolated from the stem and leaves of Anstolochia contorta Bunge, together with 17 known compounds (3-19). The structures of these compounds were determined by spectroscopic analysis. The phenanthrenes obtained were tested for cytotoxicity against renal proximal tubular epithelial cell line (HK-2). Aristololactam IVa and 7-methoxy aristololactam IV were found to have strong cytotoxic activity against HK-2 cells with a potency similar to or even stronger than those of aristolochic acid I and aristololactam I.

[Application of crossed assay design in test of drug sensitivity in vitro in combined chemotherapy of liver cancer].

A cross assay design has been carried out to test the sensitivity of drugs used in combined chemotherapy of liver cancer using human tumor cloning system in vitro. The reductive rate of the tumor clone survival was synthetically compared and statistically analysed according to calculating formula of crossed assay design. It was shown that the most effective combination of chemotherapeutic scheme could be obtained by fewer testing. Moreover, comparing the tumor response to a given drug tested by the crossed assay design with that of permutation and combinations, the results were clearly correlated (P = 0.005). We would conclude that this assay shows a good reliability and deserves practical application.

VSV-MP gene therapy strategy inhibits tumor growth in nude mice model of human lung adenocarcinoma.

Vesicular stomatitis virus (VSV) matrix protein (MP) can induce in vitro apoptosis of tumor cells in the absence of other viral components. Here, the antitumor activity of VSV-MP against lung adenocarcinoma was investigated in vivo. A pVAX-plasmid DNA encoding VSV-MP and control empty vectors (pVAX) were constructed and wrapped-up with liposome. A549 and Spc-A1 human lung adenocarcinoma cells were transfected with liposomal-VSV-MP (Lip-MP) or Lip-pVAX and then examined for cell viability or apoptosis using Hoechst/propidium iodide staining by flow cytometry, and further demonstrated by caspase/poly ADP-ribose polymerase (PARP) cleavage analysis. For the in vivo study, A549 and Spc-A1 lung carcinoma models in nude mice were established and randomly assigned into three groups to receive eight 2-weekly intravenous administrations of medium alone as control, Lip-pVAX or Lip-MP, respectively. Subsequently, Lip-MP significantly reduced tumor growth and prolonged the survival of tumor-bearing mice compared with Lip-pVAX and control agents (P<0.05), with much higher apoptosis index of both in vivo and in vitro tumor cells, respectively (P<0.05). In addition, in vivo antitumoral effect was associated with natural killer-(NK) cell congregation without evidence of toxicity. These observations suggest that systemically delivering Lip-MP has a specific dual antitumor activity in human lung adenocarcinoma by inducing apoptosis and possibly stimulating NK-cell responses, it may provide a clue for developing new therapeutic approaches against human lung adenocarcinoma.

Plasmid encoding matrix protein of vesicular stomatitis viruses as an antitumor agent inhibiting rat glioma growth in situ.

AIM: Oncolytic effect of vesicular stomatitis virus (VSV) has been proved previously. Aim of the study is to investigate glioma inhibition effect of Matrix (M) protein of VSV in situ. MATERIALS AND METHODS: A recombinant plasmid encoding VSV M protein (PM) was genetically engineered, and then transfected into cultured C6 gliomas cells in vitro. C6 transfected with Liposome-encapsulated PM (LEPM) was implanted intracranially for tumorigenicity study. In treatment experiment, rats were sequentially established intracranial gliomas with wild-typed C6 cells, and accepted LEPM injection intravenously. Possible mechanism of M protein was studied by using Hoechst staining, PI-stained flow cytometric analysis, TUNEL staining and CD31 staining. RESULTS: M protein can induce generous gliomas lysis in vitro. None of the rats implanted with LEPM-treated cells developed any significant tumors, whereas all rats in control group developed tumors. In treatment experiment, smaller tumor volume and prolonged survival time was found in the LEPM-treated group. Histological studies revealed that possible mechanism were apoptosis and anti-angiogenesis. CONCLUSION: VSV-M protein can inhibit gliomas growth in vitro and in situ, which indicates such a potential novel biotherapeutic strategy for glioma treatment.

Mannan-modified adenovirus as a vaccine to induce antitumor immunity.

Tumor vaccine is a useful strategy for cancer therapy. However, priming of the immune system requires the relevant antigen to be presented by antigen-presenting cells (APCs). Here, we employed telomerase reverse transcriptase as a model antigen to explore the feasibility of using mannan-modified adenovirus as a tumor vaccine. We found that tumor immunogene therapy with the vaccine was effective at protective antitumor immunity in mice. The antigen-specific cytotoxic T lymphocytes were found in in vitro cytotoxicity assay. The elevation of the killing activity could be abrogated by anti-CD8 or anti-major histocompatibility complex-I antibodies. Adoptive transfer of purified CD8+ cells, and CD4+ cells to a less extent, was effective at antitumor activity. In vivo antitumor activity could be abrogated by depleting CD4+ T lymphocytes. A possible explanation for the antitumor effects may be the antigen was transferred to APCs in the presence of mannan. These observations provide insights into the design of novel vaccine strategies and might be important for the future application of antigens identified in other diseases.

Combination of MIG (CXCL9) chemokine gene therapy with low-dose cisplatin improves therapeutic efficacy against murine carcinoma.

MIG (monokine induced by interferon-gamma) is a CXC chemokine ligand (CXCL9) that can potently inhibit angiogenesis, and displays thymus-dependent antitumor effects. The effectiveness of a treatment combining gene therapy with plasmid-borne MIG (pORF-MIG) and low-dose cisplatin chemotherapy was determined using colon carcinoma (CT26) and Lewis lung carcinoma (LL/2c) murine models. The program was carried out via intramuscular delivery of pORF-MIG at 100 mug/mouse twice a week for 4 weeks, and/or intraperitoneal delivery of cisplatin at 0.6 mg/kg/mouse every 3 days for 48 days. Tumor volume and survival time were evaluated after treatment. CD31 immunohistochemical staining in tumor tissues and alginate capsule models in vivo was used to evaluate angiogenesis. Induction of apoptosis and cytotoxic T-lymphocyte (CTL) activity were also assessed. The combination of pORF-MIG and low-dose cisplatin produced significant antitumor activity, with complete tumor regression in 4/10 of CT26 colon carcinomas and 3/10 of LL/2c lung carcinomas, low vascularity, in alginate capsules, apparently degraded tumor microvessel density, and increased induction of apoptotic and CTL activities compared with either treatment alone. This study suggests that the combination of pORF-MIG plus cisplatin augments the inhibition of angiogenesis and the induction of apoptosis or CTL activity, all of which enhance antitumor activity. These findings may prove useful in further explorations of the application of combinatorial approaches to the treatment of solid tumors.