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The objective of this study was to investigate the potential effects from exposure to multiple metals, including cadmium (Cd), chromium (Cr), arsenic (As), mercury (Hg), nickel (Ni) and lead (Pb) that exist in shellfish in Chinese populations, and evaluate the impacts of cumulative exposures among individuals. A total 15,469 individual shellfish samples were analyzed following a protocol for elemental analysis. A total of 25,669 respondents were selected through stratified multi-stage cluster sampling from 31 survey sites in China. Individual shellfish consumption data and body weight were collected and analyzed with a 24-hour dietary recall method on three non-consecutive days. The consumers were divided into two groups: aged 2-12 years group and aged ≥13 years. Exposure levels were estimated based using a semi-probabilistic risk assessment method and cumulative exposures among individual persons using the maximum cumulative ratio (MCR) method. The mean, median and 95th percentile of all metals exposure for age 2-12 years group were higher than age ≥13 years group, but none exceeded the respective reference doses of the metals. The mean hazard indices (HI) of each metal in shellfish for both age groups were less than 1. The MCR values calculated among 1283 consumers ranged from 1.28 to 3.25 with the average MCR value was 2.22. There were 51 (3.98%) consumers in group I, 1186 (92.44%) consumers in group II, 26(2.03%) consumers in group III A, and 20 (1.56%) consumers in group III B. And cadmium (Cd) and lead (Pb) produced the HQ

Assessment of contaminated food through the dietary intake is essential for human health. To investigate the health risk of rare earth element (REE) exposure to fruits and vegetables in mining areas in China, we collected 288 fruit samples and 942 vegetable samples from four representative mining points (Bayan Obo in Inner Mongolia, Weishan in Shandong, Maoming in Guangdong, Longnan in Jiangxi) and their control areas. The content of REEs was determined by inductively coupled plasma-mass spectrometry (ICP-MS). The total REEs in fruits from mining and control areas were 12.90 μg kg

Rare earth elements (REEs) are widely found in foods. A high intake of REEs may have associations with adverse effects on human health. This study aimed to investigate the concentrations of REEs in foods in China and to assess the risk of dietary REEs exposure in the Chinese population. The mean concentrations of the total REEs in 27,457 food samples from 11 food categories ranged from 0.04 to 1.41 mg/kg. The daily mean dietary exposure of the total REEs was 1.62 μg/kg BW in the general Chinese population and ranged from 1.61 to 2.80 μg/kg BW in different sex–age groups. The high consumer exposure (95th percentile, P95) was 4.83 μg/kg BW, 9.38% of the temporary ADI (tADI) of REEs (51.5 μg/kg BW). None of the P95 exposure exceeded the tADI in all of the sub-groups. Lanthanum, cerium, and yttrium accounted for approximately 63% of the total exposure of the 16 REEs. The hazard index of 16 REEs was far below 1. Therefore, the health risk of dietary REEs exposure in the general Chinese population was low. No cumulative risk was found for the 16 REEs in China. The results indicate there was no need to stipulate the limits of REEs in foods.

Funder: French National Cancer Institute, DNA double strand breaks (DSBs) are induced by external genotoxic agents (ionizing radiation or genotoxins) or by internal processes (recombination intermediates in lymphocytes or by replication errors). The DNA ends induced by these genotoxic processes are often not ligatable, requiring potentially mutagenic end-processing to render ends compatible for ligation by non-homologous end-joining (NHEJ). Using single molecule approaches, Loparo et al. propose that NHEJ fidelity can be maintained by restricting end-processing to a ligation competent short-range NHEJ complex that 'maximizes the fidelity of DNA repair'. These in vitro studies show that although this short-range NHEJ complex requires DNA ligase IV (Lig4), its catalytic activity is dispensable. Here using cellular models, we show that inactive Lig4 robustly promotes DNA repair in living cells. Compared to repair products from wild-type cells, those isolated from cells with inactive Lig4 show a somewhat increased fraction that utilize micro-homology (MH) at the joining site consistent with alternative end-joining (a-EJ). But unlike a-EJ in the absence of NHEJ, a large percentage of joints isolated from cells with inactive Lig4 occur with no MH - thus, clearly distinct from a-EJ. Finally, biochemical assays demonstrate that the inactive Lig4 complex promotes the activity of DNA ligase III (Lig3).

Background: Chromosome instability (CIN), characterized by frequent and ongoing loss or gain of chromosome number, is commonly observed in tumor cells. Although long recognized as a vulnerability of cancer cells, potential CIN-selective therapeutic targets have only recently been discovered. Genetic studies from multiple groups have identified the mitotic kinesin, KIF18A, as selectively essential for the proliferation of CIN and aneuploid cells. By targeting KIF18A genetically and with novel small molecule inhibitors here we present data supporting KIF18A as therapeutic target in CIN tumors. Methods and Materials: To explore the therapeutic potential of KIF18A, we evaluated the effects of KIF18A genetic depletion and small molecule inhibition in both CIN-positive and CIN-negative cell lines. Biochemical, cell proliferation and phenotypic assays were used to characterize the potency and cellular activity of reference and novel KIF18A small molecule inhibitors. Anti-tumor activity of KIF18A inhibitors was assessed in CIN-positive cell line xenograft models. Results: In triple negative breast and colorectal CIN positive cancer cell lines siRNA mediated KIF18A knockdown and small molecule inhibition of the KIF18A ATPase activity both lead to a reduction in proliferation associated with an increase in mitotic index and multi-polar spindles. Consistent with KIF18A knockdown, KIF18A inhibition results in increased spindle length and chromosome alignment defects. Importantly, these effects are not observed with KIF18A knockdown or KIF18A inhibition in non-transformed, near diploid cells. In vivo, treatment of CIN-positive xenograft tumors with potent, novel KIF18A inhibitors results in robust anti-tumor activity with minimal impact on body weight. Conclusions: Collectively our data support KIF18A as a therapeutic target and provide rationale for the continued development of potent selective small molecule KIF18A inhibitors for the treatment of CIN positive cancers. Citation Format: James D. Joseph, Katherine Schutt, Kira Fisher, Katelyn Queen, Olivia Budington, Weifeng Mao, Wei Liu, Zhengqing Zhu, Xiangping Zhang, Yisong Xiao, Kunmin Lai, Xiaohui Gu, Jason Stumpff, Fred Aswad. Targeting the mitotic kinesin, KIF18A, in chromosomally unstable cancers. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4965.

To analyze the eating out of home behavior of urban adults in China.Samples were chosen from China Food Consumption Survey in 2017. A total of 17 234 participants aged 18 and above were included in the final analysis. The food frequency questionnaire were used to collect eating out of home status in the past week. χ~2 test was used to compare the difference in the rate of eating out of home and dining places among different groups. Non-parametric test was used to compare the differences in dining out times.The overall prevalence of eating out of home was 55.6% in urban adults aged in 2017. The average number of eating out of home was 2.8 times. The proportion of eating in hotels and restaurants was 36.0%. The proportion of eating in the canteen of school, workplace and other places was 19.8%. The rate of eating out of home and dining out times were higher among male, 18-44 years old, people with higher educational level and higher household income. The proportion of students eating in canteen was higher. The proportion of professional technicians and service staff eating in hotels and restaurants were higher.Eating out of home is more common among urban adults aged 18 and above in China. Young people aged 18-44 years old eat out more often in the past week. The proportion of people eating in hotels and restaurants is higher.

Radiation therapy plays a vital role in the treatment of tumours. In particular, the occurrence of the "abscopal effect" brings about a favourable turn for the treatment of patients with advanced metastatic malignant tumours. Because of the abscopal effect, non-irradiated areas are also treated. However, the abscopal effect occurs by chance, not through seeking. Although the abscopal effect has been studied enthusiastically, the desired result does not appear to be achieved. Moreover, its combination with immunotherapy appears to be overwhelming. There is an opinion that abscopal effect is difficult to achieve by irradiation of a single tumour, and irradiation of multiple or total lesions is advocated to increase the possibility of obtaining clinically meaningful outcomes. Obviously, there are still questions about the mechanism, condition and possibility underlying the occurrence of the abscopal effect. Can the abscopal effect truly change the future treatment strategy as the researchers expect? What are the current problems? This article reviewed the research in recent years to explore the progress and controversy surrounding the abscopal effect of radiation therapy.

The economical production of biodiesel from lignocellulose requires an effective co-fermentation of lignocellulose-derived sugars, such as glucose and xylose. Our present work discovered that Lipomyces starkeyi AS 2.1560 could produce microbial lipid effectively from co-fermentation of glucose and xylose in a two-stage culture mode under non-sterile conditions. High lipid production was achieved by a feed-batch fermentation process without sterility in the second fermentation stage. After incubation for 46 h in a bioreactor utilizing unsterile glucose and xylose solution, the cell biomass, lipid and lipid content reached 98.3 g L−1, 62.7 g L−1 and 63.8%, respectively. The results showed that the two-stage fermentation method should be a promising process to produce lipid from lignocellulose hydrolysates.

Pregnant women are susceptible to adverse health effects associated with phthalate acid esters (PAEs) and polycyclic aromatic hydrocarbons (PAHs), and diet is a significant exposure source. Little is known about the contributions of dietary patterns during pregnancy to the exposure variability of these environmental contaminants.To identify dietary patterns in relation to PAEs and PAHs exposure in the Chinese pregnant population.Dietary data and urinary concentrations of environmental pollutants were obtained from 1190 pregnant women in the Tongji Birth Cohort (TJBC). PAEs and PAHs were measured in spot urine samples. Food intake was assessed using a food-frequency questionnaire. Dietary patterns were constructed by principal component analysis (PCA). Through PCA, we also extracted three chemical mixture scores that represent different co-exposure patterns of PAEs and PAHs. Multiple linear regression models were adopted to identify predictors of PAEs and PAHs exposure.Four dietary patterns were identified by PCA that explained 44.9 % of the total variance of food intake. We found egg-dairy products pattern, whole grain-tuber crop pattern, and meat-aquatic products pattern were positively associated with specific pollutants exposure. In contrast, fruit-nut-vegetable pattern was negatively correlated with PAEs and PAHs exposure. Every SD increase in this pattern score was associated with 14.36 % reduced mono(2-ethyl-5-oxohexyl) phthalate (MEOHP) (95 % CI: -24.50 ~ -2.96, p-trend = 0.01), 10.86 % reduced 2-hydroxynaphthalene (2-OHNap) (95 % CI: -20.07 ~ -0.60, p-trend = 0.04), 19.35 % reduced 9-hydroxyphenanthrene (9-OHPhe) (95 % CI: -34.49 ~ -0.70, p-trend = 0.01), and 8.33 % reduced scores of PAHs group (95 % CI: -15.97 ~ -0.10, p-trend = 0.02). In addition, disposable tableware usage and passive smoking were suggested as potentially modifiable sources of PAEs and PAHs exposure, respectively.Adhering to egg-dairy products pattern, whole grain-tuber crop pattern, and meat-aquatic products pattern may be related to increased PAEs and PAHs exposure, while following fruit-nut-vegetable pattern seems to correlate with a lower burden of such exposure.

Background Rare earth element (REE) have been considered to have potential effects on human health. However, little is known about the health risk of REEs exposure among tea drinkers in China. Objectives To characterize the concentration of REEs in different categories of tea and assess the health risk of REEs of tea consumers in China. Methods 4326 tea samples(including oolong tea, dark tea, black tea, white tea, green tea and flowering tea from markets of 24 provinces, autonomous regions and municipalities in China, were analyzed for sixteen kinds of rare earth elements. Combining the REE concentration data with tea consumption data, derived from the China National Food Consumption Survey 2013–2014, the REE exposure level through tea for adults was estimated. Calculate the hazard quotient, and judge the health risk of the population based on the result value Results The average concentrations of rare earth elements in Oolong tea and black tea were the highest (1611.8µg/kg and 1435.2µg/kg), and in green tea and flower tea were the lowest (735.1µg/kg and 564.1µg/kg). The content values of light rare earth elements (LREEs) were dominant in 94.53% of tea samples, cerium (431.5µg/kg) was the most abundant, followed by lanthanum (267.3µg/kg). The average daily dose of REEs in tea intake for Chinese adults was 0.0328µg/kg BW. There are statistical differences in REEs exposure among different genders of tea drinkers(p

The identification and lead optimization of a series of pyrazolo[3,4-d]pyridazinone derivatives are described as a novel class of potent irreversible BTK inhibitors, resulting in the discovery of compound 8. Compound 8 exhibited high potency against BTK kinase and acceptable PK profile. Furthermore, compound 8 demonstrated significant in vivo efficacy in a mouse-collagen-induced arthritis (CIA) model.

Berberine (BBR) is a natural isoquinoline alkaloid, which is used in traditional medicine for its anti‐microbial, anti‐protozoal, anti‐diarrhoeal activities. Berberine interacts with DNA and displays anti‐cancer activities, yet its effects on cellular DNA repair and on synthetic treatments with chemotherapeutic drugs remain unclear. In this study, we investigated the effects of BBR on DNA repair and on sensitization of breast cancer cells to different types of DNA damage anti‐tumoural drugs. We found BBR arrested cells in the cell cycle S phase and induced DNA breaks. Cell growth analysis showed BBR sensitized MDA‐MB‐231 cells to cisplatin, camptothecin and methyl methanesulfonate; however, BBR had no synergistic effects with hydroxurea and olaparib. These results suggest BBR only affects specific DNA repair pathways. Western blot showed BBR down‐regulated XRCC1 expressions, and the rescued XRCC1 recovered the resistance of cancer cells to BBR. Therefore, we conclude that BBR interferes with XRCC1‐mediated base excision repair to sensitize cancer cells to chemotherapeutic drugs. These finding can contribute to understanding the effects of BBR on cellular DNA repair and the clinical employment of BBR in treatment of breast cancer.

Background Malathion has been wildly used in agriculture and household pest control for years and can affect human health through drinking water and dietary exposure. This study was to investigate the immunotoxicity of malathion (MLT）on BALA/c mice for the purpose of providing a scientific basis for pesticides registration management of MLT and other pesticides. Methods Female mice were randomly divided into five groups. Both the negative control and the positive control groups of mice were daily gavage with corn oil, and those in the positive control group were intraperitoneal injected with 200 mg/kg bw of cyclophosphamide (CY) 24 h before the end of the study. In the dosage groups, MLT were daily gavage at doses of 16, 65 and 258 mg/kg bw for 30 days, respectively. After exposure, the body weight, the absolute and relative weight of organs, the hematology and clinical chemistry parameters, the splenocyte number, the bone marrow cell classification, histopathology, as well as the cellular immunity, humoral immunity, and non-specific immunity functions of mice were studied. Comparisons between multiple groups were carried out by using one-way ANOVA followed by Bonferroni post hoc comparisons tests when equal variances assumed and Dunnett’s T3 post hoc tests when equal variances assumption was not met.ResultsCompared with the negative control group, the low dose of MLT significantly reduced the levels of IL-10, neutrophil, polychromatic erythroblast and monocyte series (PPP

This study was aimed to evaluate the toxic effects of different concentrations (23, 90, 360 mg/kg BW) of atrazine (ATZ) on immune function in BALB/c mice. Some parameters of general immunotoxicity, humoral immunity, cellular immunity, and non-specific immunity were tested. The studies showed that the high-dose ATZ induced a significant reduction in the final body weight of mice, the absolute and relative weights of spleen, the counts of white blood cell (WBC), lymphocyte (LYM), monocyte (MON), and the number of splenocyte. An increase in the level of alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and cholesterol (CHO) in the high-dose ATZ group was observed. Pathological examination showed that the medium- and high-doses of ATZ caused atrophy and destruction of thymus, spleen, and hepatorenal toxicity. The serum interleukin-5(IL-5) level of mice and the number of plaque-forming cell (PFC) in spleen cells in the high-dose ATZ group decreased significantly while there was a significant increase of the serum immunoglobulin G (IgG) in the high-dose ATZ group when compared to the negative control group. In the high-dose ATZ group, the proliferation ability of T and B lymphocytes as well as the delayed-type hypersensitivity (DTH) response were significantly decreased. The low-dose ATZ (23 mg/kg BW) caused a significant decrease in the number of WBC and neutrophil (NEUT), as well as the proportion of polychromatic and normoblast. In summary, we thought the low-dose ATZ has a slight effect on the immune system; it can be preliminarily concluded that the lowest observed adverse effect level (LOAEL) of atrazine is 23 mg/kg BW in mice. Atrazine can cause immunotoxicity mainly through cellular and humoral immunity pathways.

Berberine (BBR) is an isoquinoline alkaloid isolated from Cotridis rhizoma and exhibits multiple biological roles including anti-microbe, anti-inflammation and anti-tumor activities. In this study, two triple-negative breast cancer cell (TNBC) lines, MDA-MB-231 and BT549, were used to investigate the effect of BBR on growth of TNBC in vitro and in vivo. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate the viability of cells treated with BBR. After 48h treatments, a 50% inhibitory concentration (IC50) of BBR to BT549 and MDA-MB-231 cells are at 16.575±1.219μg/ml and 18.525±6.139μg/ml respectively. BBR reduced colony formation of BT549 and MDA-MB-231 cells. The wound-healing assay showed BBR decreased breast cancer cell migrations (P

PTEN is a tumor suppressor gene well characterized as a phosphatase. However, more evidences demonstrate PTEN functions in DNA repair independent of its phosphatase activity, which affects the efficacy of DNA damage anti-tumoral drugs in treating cancer cells with PTEN variations. Using BT549 breast cancer cells, we studied the roles of PTEN in DNA repair and in sensitization of breast cancer cells to olaparib, a poly(ADP-ribose) polymerase (PARP) inhibitor. Comet assay showed PTEN promoted DNA repair. PTEN-deficient BT549 cells are sensitive to olaparib, which shows the synthetic lethality between PTEN and PARP1. We expressed PTEN in BT549 cells and found PTEN-proficient BT549 cells resist to olaparib. Western blot showed that PTEN up-regulated Rad51 expression, suggesting PTEN promotes DNA repair through Rad51-dependnent homologous recombination. We used 5μM olaparib or 5μM RI-1, a Rad51 inhibitor, to treat PTEN-proficient BT549 cells respectively. The immunofluorescent analysis showed the combination of olaparib and RI-1 induced more than 4-fold of γH2AX foci than either of them. MTT assay showed 5μM RI-1 did not change the survival of PTEN-proficient BT549 cells, however, this dose of RI-1 sensitized PTEN-proficient BT549 cells to olaparib. Consequently, these results demonstrate that inhibition of Rad51 can sensitize BT549 cells with wild type PTEN to olaparib, which would contribute to using PARP inhibitors in individual treatment of breast cancer patients with PTEN variations.

Triptolide is the primary compound isolated from Tripterygium wilfordii, which has been reported to inhibit nucleotide excision repair as well as exhibit anti-inflammatory and antitumor activities. However, the action of triptolide in DNA breaks remains unknown. The present study investigated the effects of triptolide in the induction of DNA breaks and apoptosis in a murine B-cell lymphoma cell line, CH12F3. An MTT assay revealed that X-ray repair cross-complementing protein 1 (XRCC1)−/− CH12F3 cells were more sensitive to 6 nM triptolide compared with the wild-type CH12F3 cells, which suggests that low levels of triptolide induce DNA breaks in a manner that is dependent on the XRCC1-mediated repair pathway. Flow cytometric analysis identified that 50 nM triptolide increased the phospho-histone H2AX level, demonstrating that triptolide induces double-strand breaks. Western blot analysis revealed that triptolide up-regulated Rad51 and nuclear proliferating cell nuclear antigen. Annexin V/propidium iodide staining identified that triptolide promoted apoptosis and western blot analysis confirmed that triptolide activated caspase-3-dependent apoptosis. The results of the present study also demonstrated that 5 nM triptolide sensitized CH12F3 lymphoma cells to poly(ADP-ribose) polymerase 1 and phosphoinositide 3-kinase inhibitors, suggesting that triptolide may be a potent antitumor drug for sensitizing lymphoma cells to chemotherapeutic agents.

A high-throughput screening (HTS) of the Genentech/Roche library identified a novel, uncharged scaffold as a KDM5A inhibitor. Lacking insight into the binding mode, initial attempts to improve inhibitor potency failed to improve potency, and synthesis of analogs was further hampered by the presence of a C-C bond between the pyrrolidine and pyridine. Replacing this with a C-N bond significantly simplified synthesis, yielding pyrazole analog 35, of which we obtained a co-crystal structure with KDM5A. Using structure-based design approach, we identified 50 with improved biochemical, cell potency and reduced MW and lower lipophilicity (LogD) compared with the original hit. Furthermore, 50 showed lower clearance than 9 in mice. In combination with its remarkably low plasma protein binding (PPB) in mice (40%), oral dosing of 50 at 5mg/kg resulted in unbound Cmax ∼2-fold of its cell potency (PC9 H3K4Me3 0.96μM), meeting our criteria for an in vivo tool compound from a new scaffold.

Introduction of a Michael acceptor on a flexible scaffold derived from pan-FGFR inhibitors has successfully yielded a novel series of highly potent FGFR4 inhibitors with selectivity over FGFR1. Due to reduced lipophilicity and aromatic ring count, this series demonstrated improved solubility and permeability. However, plasma instability and fast metabolism limited its potential for in vivo studies. Efforts have been made to address these problems, which led to the discovery of compound (-)-11 with improved stability, CYP inhibition, and good activity/selectivity for further optimization.

[Image: see text] The identification and lead optimization of a series of pyrazolo[3,4-d]pyridazinone derivatives are described as a novel class of potent irreversible BTK inhibitors, resulting in the discovery of compound 8. Compound 8 exhibited high potency against BTK kinase and acceptable PK profile. Furthermore, compound 8 demonstrated significant in vivo efficacy in a mouse-collagen-induced arthritis (CIA) model.

Context: Atherosclerosis is a chronic inflammatory disease in which the plaques were built up inside of the artery. Interleukin-8 (IL-8, CXCL8) is an inflammatory factor, known to play an important...

Pancreatic cancer is devastating cancer worldwide with few if any truly effective therapies. Pancreatic cancer has an increasing incidence and may become the second leading cause of death from cancer. Novel, more effective therapeutic approaches are needed as pancreatic cancer patients usually survive for less than a year after being diagnosed. Control of blood sugar levels by the prescription drug metformin in diseases such as diabetes mellitus has been examined in association with pancreatic cancer. While the clinical trials remain inconclusive, there is hope that certain diets and medications may affect positively the outcomes of patients with pancreatic and other cancers. Other natural compounds may share some of the effects of metformin. One “medicinal” fruit consumed by millions worldwide is berberine (BBR). Metformin and BBR both activate AMP-activated protein kinase (AMPK) which is a key mediator of glucose metabolism. Glucose metabolism has been shown to be very important in cancer and its significance is increasing. In the following studies, we have examined the effects of metformin, BBR and a panel of modified BBRs (NAX compounds) and chemotherapeutic drugs on the growth of four different human pancreatic adenocarcinoma cell lines (PDAC). Interestingly, the effects of metformin could be enhanced by BBR and certain modified BBRs. Upon restoration of WT-TP53 activity in MIA-PaCa-2 cells, an altered sensitivity to the combination of certain NAX compounds and metformin was observed compared to the parental cells which normally lack WT-TP53. Certain NAX compounds may interact with WT-TP53 and metformin treatment to alter the expression of key molecules involved in cell growth. These results suggest a therapeutic approach by combining certain pharmaceutical drugs and nutraceuticals to suppress the growth of cancer cells.

To investigate the effect of CXCL12 on regeneration of radial glia like cells after traumatic brain injury (TBI). We randomly divided 48 rats into 4 groups: (1) the sham group, rats were performed craniotomy only, (2) the control group, saline were injected into the ipsilateral cortex after TBI, (3) the CXCL12 group, CXCL12 were injected, and (4) the CXCL12 + AMD3100 group, a mixture of CXCL12 and AMD3100 were injected. Seven days after TBI, the brain tissues were subjected to immunofluorescence double-labeled staining of BrdU/Nestin, BLBP/Nestin, BLBP/Vimentin, BLBP/SOX2, BLBP/CXCR4, BLBP/DCX. Western Blot assay was used to measure the levels of Nestin, BLBP, and Vimentin. Compared with the control group, CXCL12 treatment significantly increased the number of cells stained with BrdU/Nestin, BLBP/Nestin, and BLBP/Vimentin around the injured cortex and corpus callosum areas. CXCL12 + AMD3100 treatment significantly decreased the number of these cells compared with the CXCL12 treatment and control group. The protein levels of Nestin, BLBP, and Vimentin had the same change trends as those of the immunofluorescence staining. The BLBP/Vimentin positive cells presented with the astrocyte pattern around the injured cortex area but with the RGCs pattern around the injured corpus callosum area. The BLBP positive cells also expressed CXCR4 and SOX2. Altogether, CXCL12 promotes the proliferation of neural precursor cells after TBI by combing to its receptor, CXCR4. The proliferating neural precursor cells presents radial glial cell like cells. The RGCs-like cells can differentiate into immature neurons and promote the migration of immature neurons.

Mutations at the TP53 gene are readily detected (approximately 50–75%) in pancreatic ductal adenocarcinoma (PDAC) patients. TP53 was previously thought to be a difficult target as it is often mutated, deleted or inactivated on both chromosomes in certain cancers. In the following study, the effects of restoration of wild-type (WT) TP53 activity on the sensitivities of MIA-PaCa-2 pancreatic cancer cells to the MDM2 inhibitor nutlin-3a in combination with chemotherapy, targeted therapy, as well as, nutraceuticals were examined. Upon introduction of the WT-TP53 gene into MIA-PaCa-2 cells, which contain a TP53 gain of function (GOF) mutation, the sensitivity to the MDM2 inhibitor increased. However, effects of nutlin-3a were also observed in MIA-PaCa-2 cells lacking WT-TP53, as upon co-treatment with nutlin-3a, the sensitivity to certain inhibitors, chemotherapeutic drugs and nutraceuticals increased. Interestingly, co-treatment with nutlin-3a and certain chemotherapeutic drug such as irinotecan and oxaliplatin resulted in antagonistic effects in cells both lacking and containing WT-TP53 activity. These studies indicate the sensitizing abilities that WT-TP53 activity can have in PDAC cells which normally lack WT-TP53, as well as, the effects that the MDM2 inhibitor nutlin-3a can have in both cells containing and lacking WT-TP53 to various therapeutic agents.

Berberine (BBR) is a common nutraceutical consumed by millions worldwide. BBR has many different effects on human health, e.g., diabetes, diarrhea, inflammation and now more recently it has been proposed to have potent anti-cancer effects. BBR has been shown to suppress the growth of cancer cells more than normal cells. BBR has been proposed to exert its growth-inhibitory effects by many different biochemical mechanisms including: suppression of cell cycle progression, induction of reactive oxygen species, induction of apoptosis and autophagy and interactions with DNA potentially leading to DNA damage, and altered gene expression. Pancreatic cancer is a leading cancer worldwide associated with a poor prognosis. As our population ages, pancreatic cancer has an increasing incidence and will likely become the second leading cause of death from cancer. There are few truly-effective therapeutic options for pancreatic cancer. Surgery and certain chemotherapeutic drugs are used to treat pancreatic cancer patients. Novel approaches to treat pancreatic cancer patients are direly needed as they usually survive for less than a year after being diagnosed. In the following manuscript, we discuss the abilities of BBR and certain chemically- modified BBRs (NAX compounds) to suppress growth of pancreatic cancer cells.

Efficient and economical production of renewable fuels and chemicals requires the co-utilization of sugars derived from lignocellulosic biomass as feedstock. Herein, Rhodosporidium toruloides cells were employed to produce microbial lipid via co-fermentation of lignocellulosic biomass-derived non-sterile sugars. In this study, we found that Rhodosporidium toruloides was very effective lipid biosynthesis yeast for co-fermenting of lignocellulosic biomass-derived sugars through the two-phase whole-cell biocatalytic process. Moreover, under the two-phase biosynthesis condition, the lipid production of non-sterile fermentation was higher than that of sterile fermentation. Accordingly, the two-phase whole-cell biocatalytic process for lipid biosynthesis can reduce the energy consume for the sterilization. Especially high lipid productivity was achieved from mixed sugars solution without sterility in bioreactor. After incubation for 47 h in bioreactor, the biomass, lipid and lipid contents were achieved in 56.8 gL-1, 92.7 gL-1 and 61.3%, respectively. Co-fermenting of lignocellulosic biomass-derived sugars may reduce the cost of lipid production. Fermentation of non-sterile condition can further decrease the cost of sterilization steam and cooling water by 357 RMB/ton (about 55 USD/ton). The composition of fatty acid was consistent with common plant oils and was suitable as feedstock for biodiesel production. The results of the present work show that the two-phase whole-cell biocatalytic process has a great potential in industrial lipid biosynthesis for biodiesel production.

Starting with a lead [1,5-a]pyrimidin-7(4H)-one-containing molecule (1), we generated potent, selective and orally bioavailable KDM5 inhibitors. Using structure- and property-based approaches, we designed 48 with improved cell potency (PC9 H3K4Me3 EC50=0.34μM). Furthermore, 48 maintained suitable physiochemical properties and displayed an excellent pharmacokinetic (PK) profile in mice. When dosed orally in mice at 50mg/kg twice a day (BID), 48 showed an unbound maximal plasma concentration (Cmax) >15-fold over its cell EC50, thereby providing a robust chemical probe for studying KDM5 biological functions in vivo.

To investigate the effect of CXCL12 on migration of neural precursor cells after traumatic brain injury (TBI). We randomly divided 48 rats into four groups: (1) the sham group, rats were performed craniotomy only, (2) the control group, saline were injected into the ipsilateral cortex after TBI, (3) the CXCL12 group, CXCL12 were injected into the ipsilateral cortex after TBI, and (4) the CXCL12 + AMD3100 group, CXCL12 and AMD3100 were mixed together and injected into the ipsilateral cortex after TBI. At 7 days after TBI, the brain tissues were subjected to immunofluorescent double-labeled staining with the antibodies of CXCR4/DCX, MMP-2/DCX, MMP-2/GFAP, MMP-2/NeuN. Western blot assay was used to measure the protein levels of MMP-2. Compared with the control group, the number of CXCR4/DCX and MMP-2 positive cells around the injured corpus callosum area were significantly increased in the CXCL12 treatment group. The area occupied by these cells expanded and the shape changed from chain distribution to radial. CXCL12 + AMD3100 treatment significantly decreased the number and distribution area of CXCR4/DCX and MMP-2 positive cells compared with the CXCL12 treatment and control group. The DCX positive cells could not form chain or radial distribution. The protein expressions of MMP-2 had the similar change trends as the results of immunofluorescent staining. MMP-2 could be secreted by DCX, GFAP and NeuN positive cells. CXCL12/CXCR4 axis can improve the migration of the neuroblasts along the corpus callosum by stimulating the MMP-2 secretion of different types of cells.

Radiation is a current standard treatment of glioma. The fractionated radiotherapy with low dose of radiation over weeks has been employed in glioma patients, while radiotherapy can only offer palliation due to the radioresistance. We cumulatively radiated a glioblastoma cell line, U87MG, and screened radioresistant glioma cells. A transcriptome sequencing was performed to analyze the transcription differences between the raidoresistant and control cells, which showed the mitochondria NADH–ubiquinone oxidoreductase (Complex I) subunits were up-regulated in the radioresistant cells. The copy numbers of mitochondria were increased in the radioresistant glioma cells. After using mitochondria Complex I inhibitors, rotenone and metformin, to treat glioma cells, we found the resistant glioma cells re-sensitized to radiation. These results demonstrate that Complex I is associated with the fractioned radiation-induced radioresistance of glioma and would be a potent target for clinical radiotherapy of glioma.

Glycogen synthase kinase-3β (GSK-3β) is an evolutionarily conserved serine/threonine kinase, functioning in numerous cellular processes including cell proliferation, DNA repair, cell cycle, signaling and metabolic pathways. GSK-3β is implicated in different diseases including inflammation, neurodegenerative disease, diabetes and cancers. GSK-3β is involved in biological processes of tumorigenesis, therefore, it is rational that GSK-3β inhibitors were employed to target malignant tumors. The effects of GSK-3β inhibitors in combination of radiation and chemotherapeutic drugs have been reported in various types of cancers, suggesting GSK-3β would play important roles in cancer treatments. GSK-3β is involved in multiple signal pathway including Wnt/β-catenin, PI3K/PTEN/AKT and Notch. GSK-3β also functions in DNA repair through phosphorylation of DNA repair factors and affecting their binding to chromatin. This review focuses on the molecular mechanism of GSK-3β in DNA repair, special in base excision repair and double-strands break repair, the roles of GSK-3β in inhibition of apoptosis through activation of NF-κB, and the effects of GSK-3β inhibitors on radio- and chemosensitization of various types of cancers. This article is part of a Special Issue entitled: GSK-3 and related kinases in cancer, neurological and other disorders edited by James McCubrey, Agnieszka Gizak and Dariusz Rakus.

Triptolide is a natural compound isolated from the Tripterygium wilfordii, which possesses anti-inflammatory and anti-tumor activities. Triptolide reportedly inhibits RNA polymerase II-mediated transcription and ATM activities to interfere with DNA repair. However, the roles of triptolide in DNA repair are still largely unknown. Triple negative breast cancer cells (TNBC) are insensitive to targeted anti-tumoral drugs, thus DNA damage chemotherapeutic drugs are the available treatments used in clinic, while the drug resistance of TNBC causes the challenge for successful cure. In this study, we investigated the efficiency of cisplatin in combination with triptolide in treatment of TNBC. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay shows triptolide suppresses the growth of two triple-negative breast cancer cells, BT549 and MDA-MB-231. Triptolide induces DNA breaks and arrests TNBC in the cell cycle S phase, and sensitizes TNBC to cisplatin. Western blot analysis shows triptolide down-regulated the levels of PARP1 and XRCC1, and slightly decreases the levels of RAD51. The results demonstrate triptolide interferes with single strand-break and base excision repair. The over-expressed PARP1/XRCC1 help the TNBC to resist triptolide. Based on these results, we conclude triptolide confers sensitization of TNBC to cisplatin via interference with XRCC1/PARP1-mediated base excision repair.

Curcumin is a natural compound isolated from the rhizome of Curcuma longa. It possesses anti-tumor activity through arresting cell cycles and promoting cell apoptosis. However, the effect of curcumin on DNA damage is not well defined. In this study, we investigated the effect of curcumin on inducing DNA damage and on sensitizing lymphoma cells to anti-tumoral DNA damage drugs. Western blot showed curcumin induced γ-H2AX foci in CH12F3 lymphoma cells, which suggests curcumin induces DNA breaks. In addition, curcumin decreased the expression of Rad51, which suggests curcumin induces DNA damage through regulating Rad51-dependant homologous recombination. Rad51-dependant homologous recombination is a vital DNA repair pathway for cancer cells to resist anti-tumoral DNA damage drugs, therefore, we studied the effect of curcumin on the sensitizing lymphoma cells to various chemotherapeutic drugs. We found low level of curcumin (5μM) sensitized lymphoma cells to anti-tumoral DNA damage agents including cisplatin, methyl methanesulfonate, hydroxyurea and camptothecin. We also found curcumin sensitized CH12F3 lymphoma cells to DNA-PK and PARP inhibitors. Flow cytometry analysis showed curcumin promoted apoptosis and western blot analysis confirmed curcumin activated caspase3-dependent apoptosis. Taken together, these results demonstrate that curcumin induces DNA damage through regulating Rad51-dependant homologous recombination and triggers caspase3-dependent apoptosis, more importantly, curcumin sensitizes lymphoma cells to various DNA damage drugs. Consequently, curcumin would be a potent agent for sensitizing lymphoma cells to anti-tumoral chemotherapeutic agents.

Cadmium, a toxic heavy metal, is widely present in food. It has been reported that chronic cadmium exposure is associated with kidney disease, osteoporosis, cardiovascular disease and cancer. The aim of this study was to assess the dietary cadmium exposure and potential health risk in different age-sex groups of the Chinese population (children aged 4–11 years, young people aged 12–17 years and adults aged over 18 years), and in the southern and northern population using a semi-probabilistic method. Cadmium was detected in 228,687 food samples from 32 food categrories by graphite furnace atomic absorption spectrometry. The dietary cadmium exposures were estimated by combining the cadmium concentration data with food consumption data derived from the China National Nutrient and Health Survey 2002, and evaluated against the Provisional Tolerable Monthly Intake (PTMI) of 25 μg/kg BW/month established by the Joint FAO/WHO expert committee on food additives (JECFA). The mean dietary cadmium exposure of the general Chinese population (15.3 μg/kg BW/month) was below the PTMI. The high consumer exposures (95th percentile, P95) for the general population and different sub-groups were higher than the PTMI. The dietary cadmium exposure of the southern population was apparently higher than that of the northern population. Rice was the most important contributor to cadmium exposure for Chinese people, especially those living in the southern areas of China. These findings indicated that the health risk from dietary cadmium exposure of the general Chinese people was low, but the health risk of cadmium exposure of certain sub-groups should be of concern.

Objective: Non-small-cell lung cancer (NSCLC) patients with epidermal growth factor receptor-activating mutations (EGFRm+) have a much higher risk of developing CNS metastases. The cumulative incidence of BM and LM in these patients is more than 50%. These patients have poor prognosis as there are no approved drugs for these indications. Currently approved drugs can neither effectively treat brain metastases nor prevent development of the brain metastases due to their limited blood-brain-barrier (BBB) penetration. TUL-001 is a potent oral EGFR inhibitor, which has excellent potency and BBB (blood brain barrier) penetration ability, it can potently induces tumor regression both in the brain metastases and subcutaneous animal models. Methods: The anti-proliferative activity of TUL-001 was evaluated in PC-9 (EGFRExon19 Del), and A431 (EGFRWT) NSCLC cell lines. The anti-tumor activity was evaluated in PC-9-luc Lung cancer brain metastases model, which was monitored by an IVIS Xenogen imaging system. The in vivo anti-tumor activity also evaluated in PC-9 subcutaneous transplantation tumor model. Effects on EGFR signaling pathway were assessed by western blot analysis of the downstream effector proteins p-EGFR (Y1068), p-AKT (Ser473) and p-ERK (Thr202/Tyr204). Results: TUL-001 displayed potent anti-proliferative activity in the EGFR mutant cell line PC-9 (IC50 =3.74 nM), and less potent activity in the A431 (EGFRWT) cell line (IC50 = 54.1 nM), it has about 14 fold selectivity for EGFRExon19 Del over EGFRWT. TUL-001 significantly inhibited tumor growth at all doses tested in the PC-9-luc brain metastases model, and significantly prolong the overall survival for the test animals. In the PC-9 subcutaneous transplantation tumor model, TUL-001 can also shrinkage the tumors, and the TGI for the lower dose group (2.5mpk) was more than 120%. Western blot analysis of the tumor samples in the brain metastases model study showed that TUL-001 inhibited p-EGFR (Y1068), p-AKT (Ser473), and p-ERK (Thr202/Tyr204), indicating that the tumor growth inhibition was through inhibition of the resistant mutant EGFR. Conclusion: TUL-001 is a potent brain penetrant EGFR inhibitor, and showed excellent antitumor activity in preclinical tumor models. These results warrant TUL-001 going to the clinic study for exploration of its potential for treating EGFRm+ NSCLC cancer with and without brain metastasis. Citation Format: Xile Liu, Liwei Mu, Bin Liu, Shouting Wu, Degang Wang, Lu Zhang, Charles Z Ding, Lihong Hu, Weifeng Mao, Zhenzhen Zhu, Chen Chen, Yuanfeng Xia, Chichung Chan, Shuhui Chen. Preclinical evaluation of TUL-001, an EGFR inhibitor for the treatment of NSCLC with brain metastasis (BM) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1316.

CXCL12 and its physiologic receptor CXCR4 are involved in controlling cell survival, proliferation and migration in adult tissues. This study aimed to investigate the effects of CXCL12 on cortical neuron apoptosis in rats after traumatic brain injury (TBI) and the potential mechanisms involved. At 3 days after TBI, in situ terminal transferase d-UTP nick-end labeling assay (TUNEL) showed that the apoptotic index (AI) deceased significantly in the CXCL12 treatment group compared with the control group (p < 0.05). Immunofluorescence double-labeled staining revealed that most of the TUNEL positive cells were NeuN positive neurons. The change trends of active caspase-3 expression were similar as those of the AI. The Bcl-2:Bax ratio was upregulated in the CXCL12 group compared with the control group. However, the effect of CXCL12 could be partially reverted by the additional use of AMD3100 (a kind of antagonist of CXCR4) (p < 0.05). Our results indicated that after TBI in rats CXCL12 combing CXCR4 receptors could inhibit the caspase-3 pathway by upregulating Bcl-2:Bax ratio, which protect neurons from apoptosis.

PTEN is a tumor suppressor gene characterized as a phosphatase that antagonizes the phosphatidylinositol 3-kinase signaling pathway in the cytoplasm. Nuclear PTEN plays roles in chromosomal stability, in which the double-strand breaks (DSB) repair mediated by homologous recombination (HR) and non-homologous end joining (NHEJ) is critical. Herein, the role of nuclear PTEN in DSB repair and the underlying molecular mechanism was investigated in this study. Using human breast cancer BT549 and MDA-MB-231 cell lines, we reveal a specific feature of PTEN that controls poly(ADP-ribosyl)ation of Ku70 and interferes with binding of Ku70 at DSB. Plasmid-based end joining and reporter assays showed that nuclear PTEN restrained NHEJ efficacy. Electrophoretic mobility shift assays showed that nuclear PTEN impaired Ku70 complex binding to DSB by 3-fold. Co-immunoprecipitation assay showed PTEN regulated poly(ADP-ribosyl)ation of Ku70 instead of directly interacting with Ku70, while PTEN promoted the poly(ADP-ribosyl)ation of PARP1 and induced the degradation of PARP1 in PTEN-WT cells exposed to DSB agents. Of note, the role of PTEN in DSB repair mostly depends on its nuclear localization rather than its phosphatase activity. As a result, the absence of nuclear PTEN rather than phosphatase-negative PTEN confers cell hypersensitivity to anti-tumor DNA damage drugs. This finding contributes to understanding the effect of PTEN in repair of DSB and using defined anti-tumor DSB drugs to treat tumor cells with aberrant PTEN.

This communication describes the identification and optimization of a series of pan-KDM5 inhibitors derived from compound 1, a hit initially identified against KDM4C. Compound 1 was optimized to afford compound 20, a 10 nM inhibitor of KDM5A. Compound 20 is highly selective for the KDM5 enzymes versus other histone lysine demethylases and demonstrates activity in a cellular assay measuring the increase in global histone 3 lysine 4 tri-methylation (H3K4me3). In addition compound 20 has good ADME properties, excellent mouse PK, and is a suitable starting point for further optimization.

DNA double-strand breaks (DSBs) are essential intermediates in Ig gene rearrangements: V(D)J and class switch recombination (CSR). In contrast to V(D)J recombination, which is almost exclusively dependent on nonhomologous end joining (NHEJ), CSR can occur in NHEJ-deficient cells via a poorly understand backup pathway (or pathways) often termed alternative end joining (A-EJ). Recently, several components of the single-strand DNA break (SSB) repair machinery, including XRCC1, have been implicated in A-EJ. To determine its role in A-EJ and CSR, Xrcc1 was deleted by targeted mutation in the CSR proficient mouse B-cell line, CH12F3. Here we demonstrate that XRCC1 deficiency slightly increases the efficiency of CSR. More importantly, Lig4 and XRCC1 double-deficient cells switch as efficiently as Lig4-deficient cells, clearly indicating that XRCC1 is dispensable for A-EJ in CH12F3 cells during CSR.

Objective: ALK is a tyrosine kinase belonging to the insulin receptor super-family. The constitutively active ALK fusions are oncogenic and linked with multiple types of human cancer. The EML4-ALK fusion has been proven to be the driver of approximately 5% of NSCLC patients. We disclose here for the first time a novel ALK inhibitor WX-0593 and its preclinical evaluation data in preclinical models of ALK driven NSCLC. Method: The enzyme activities of WX-0593 for ALKWT, ALKL1196M, ALKC1156Y and EGFRL858R/T790M were measured with corresponding kinase assays. The anti-proliferative activity was evaluated in Karpas-299, Ba/F3, Ba/F3 (EML4-ALK-WT), Ba/F3 (EML4-ALK-L1196M), Ba/F3 (EML4-ALK-C1156Y), and NCI-H1975 cell lines. The specific effect (PD) of WX-0593 on ALK signaling was analyzed in NCI-H3122 cells. Antitumor activity of WX-0593 was evaluated in three patient-derived xenograft (PDX) models of NSCLC (LU-01-0015, LU-01-0319, and crizotinib resistant LU-01-0319R) and one cell-derived xenograft (CDX) model of NSCLC (NCI-H3122). PKPD was assessed by western blot analysis of the treated tumor tissues for p-ALK (Tyr1604), p-STAT3 (Tyr705), p-STAT5 (Tyr694), p-AKT, and p-ERK. Result: WX-0593 displayed potent enzymatic activities for ALKWT, ALKL1196M, ALKC1156Y and EGFRL858R/T790M with IC50 of 5.38, 9.26, 9.28, and 16.74 nM respectively. It also showed expected anti-proliferative activity in six cell lines, Karpas-299, Ba/F3, Ba/F3 (EML4-ALK-WT), Ba/F3 (EML4- ALK-L1196M), Ba/F3 (EML4-ALK-C1156Y), and NCI-H1975, with IC50 of 4.5, 2208, 4, 9.5, 9, and 508 nM, respectively. Western blot study in NCI-H3122 cells showed that WX-0593 could fully inhibit p-ALK, p-ERK,p-STAT5 at 11.1 nM and p-AKT at 100 nM. WX-0593 significantly inhibited tumor growth at all doses tested (P Conclusion: We have identified a potent orally active second generation ALK inhibitor, WX-0593. It can inhibit the activity of both wide type and crizotinib resistant ALK and showed strong antitumor activity in in vitro and in vivo preclinical models. These results are considered highly promising and warrant moving the compound forward to clinical investigation. Citation Format: Liu Xile, Charles Z Ding, Weifeng Mao, Yuxin Qin, Shansong Zheng, Yingying Yang, Qingmei Zheng, Long Zhang, Shuyong Zhao. Preclinical evaluation of WX-0593, a potent orally active ALK inhibitor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4794.