BRCA2, PALB2, RECQL4 Germline Pathogenic Variants, and Somatic TP53 Mutation in Triple Metachronous Malignancies: A Case Report and Literature Review.

Background: Multiple primary cancer (MPC) refers to the presence of more than one cancer in an individual. Triple primary malignancies are uncommon. Case: We report the case of a 50-year-old postmenopausal woman in our gynecology department, diagnosed with endometrial cancer, ovarian cancer, and unilateral breast cancer. She carried germline mutations in BRCA2, PALB2, and RECQL4, along with a somatic pathogenic variant in TP53. Endometrial cancer patients harboring germline pathogenic variants in BRCA2 exhibit a heightened risk of ovarian and breast cancer. BRCA2 is known to play a role in the development of ovarian and breast cancer, while PALB2 is identified as a gene associated with breast cancer susceptibility. RECQL4 has been linked to breast cancer, cervical cancer, and other tumors. Conclusion: Genetic testing may be imperative for identifying MPC in endometrial cancer patients. For individuals with BRCA2 and other gene pathogenic variants, routine examination and monitoring of the endometrium, ovaries, breasts, and other sites prone to polygenic cancer are recommended.

Brain-specific loss of Abcg1 disturbs cholesterol metabolism and aggravates pyroptosis and neurological deficits after traumatic brain injury.

Based on accumulating evidence, cholesterol metabolism dysfunction has been suggested to contribute to the pathophysiological process of traumatic brain injury (TBI) and lead to neurological deficits. As a key transporter of cholesterol that efflux from cells, the ATP-binding cassette (ABC) transporter family exerts many beneficial effects on central nervous system (CNS) diseases. However, there is no study regarding the effects and mechanisms of ABCG1 on TBI. As expected, TBI resulted in the different time-course changes of cholesterol metabolism-related molecules in the injured cortex. Considering ABCG1 is expressed in neuron and glia post-TBI, we generated nestin-specific Abcg1 knockout (Abcg1-KO) mice using the Cre/loxP recombination system. These Abcg1-KO mice showed reduced plasma high-density lipoprotein cholesterol levels and increased plasma lower-density lipoprotein cholesterol levels under the base condition. After TBI, these Abcg1-KO mice were susceptible to cholesterol metabolism turbulence. Moreover, Abcg1-KO exacerbated TBI-induced pyroptosis, apoptosis, neuronal cell insult, brain edema, neurological deficits, and brain lesion volume. Importantly, we found that treating with retinoid X receptor (RXR, the upstream molecule of ABCG1) agonist, bexarotene, in Abcg1-KO mice partly rescued TBI-induced neuronal damages mentioned above and improved functional deficits versus vehicle-treated group. These data show that, in addition to regulating brain cholesterol metabolism, Abcg1 improves neurological deficits through inhibiting pyroptosis, apoptosis, neuronal cell insult, and brain edema. Moreover, our findings demonstrate that the cerebroprotection of Abcg1 on TBI partly relies on the activation of the RXRalpha/PPARgamma pathway, which provides a potential therapeutic target for treating TBI.

Interleukin-13 Affects the Recovery Processes in a Mouse Model of Hemorrhagic Stroke with Bilateral Tibial Fracture.

As one form of stroke, intracerebral hemorrhage (ICH) is a fatal cerebrovascular disease, which has high morbidity and mortality and lacks effective medical treatment. Increased infiltration of inflammatory cytokines coupled with pyroptotic cell death is involved in the pathophysiological process of ICH. However, little is known about whether concomitant fracture patients have the same progression of inflammation and pyroptosis. Hence, we respectively established the mouse ICH model and ICH with bilateral tibial fracture model (MI) to explore the potential cross-talk between the above two injuries. We found that MI obviously reversed the expressions of pyroptosis-associated proteins, which were remarkably up-regulated at the acute phase after ICH. Similar results were observed in neuronal expressions via double immunostaining. Furthermore, brain edema was also significantly alleviated in mice who suffered MI, when compared with ICH alone. To better clarify the potential mechanisms that mediated this cross-talk, recombinant mouse interleukin-13 (IL-13) was used to investigate its effect on pyroptosis in the mouse MI model, in which a lower level of IL-13 was observed. Remarkably, IL-13 administration re-awakened cell death, which was mirrored by the re-upregulation of pyroptosis-associated proteins and PI-positive cell counts. The results of hemorrhage volume and behavioral tests further confirmed its critical role in regulating neurological functions. Besides, the IL-13-treated MI group showed poor outcomes of fracture healing. To sum up, our research indicates that controlling the IL-13 content in the acute phase would be a promising target in influencing the outcomes of brain injury and fracture, and meanwhile, provides new evidence in repairing compound injuries in clinics.

[Pollution Characteristics and Risk Assessment of Nitrated Polycyclic Aromatic Hydrocarbons in the Atmosphere of Guangdong-Hong Kong-Macao Greater Bay Area].

To investigate the pollution characteristics and sources of nitrated polycyclic aromatic hydrocarbons (NPAHs) in Guangdong-Hong Kong-Macao Greater Bay Area (GBA), 44 ambient air samples were collected using the active sampling method, which were then determined via gas chromatography-triple quadrupole tandem mass spectrometry. The main results showed that filters, polyurethane foam, and XAD-2 resin were the essential materials for sampling NPAHs in ambient air in order to characterize the pollution status accurately. The levels of ρ(Σ18NPAHs) in ambient air at GBA ranged from 162 pg·m-3 to 2094 pg·m-3, and the average levels of ρ(Σ18NPAHs) were (675±430) pg·m-3 in summer and (637±349) pg·m-3 in winter. NPAHs were widely found in the ambient air of GBA and were dominated by 1-nitronaphthalene (220 pg·m-3), 2-nitronaphthalene (146 pg·m-3), 9-nitroanthracene (105 pg·m-3), and 2-nitrofluoranthene (72 pg·m-3). The congener profile characteristics of NPAHs in summer and winter were similar. The gas/particle partitioning characteristics of NPAHs revealed that dicyclic and tricyclic NPAHs tend to occur in the gas phase, and tetracyclic NPAHs tend to be adsorbed in the particle phase. The fraction of NPAHs concentrations in the particulate fraction of their total atmospheric concentrations increased with the increase in their molecular weight. In winter, NPAHs tend to be adsorbed in the particle phase, whereas in summer, NPAHs tend to exist in the gas phase. Based on the ratios of characteristic pollutants, in both the summer and winter season, photochemical reactions were the main source of NPAHs in the atmosphere of GBA and were primarily generated by the reaction of the hydroxyl radical in the daytime. The carcinogenic risk value calculation showed that the current carcinogenic risk of NPAHs in the ambient air of GBA was controllable.

Restraint Stress Delays the Recovery of Neurological Impairments and Exacerbates Brain Damages through Activating Endoplasmic Reticulum Stress-mediated Neurodegeneration/Autophagy/Apopotosis post Moderate Traumatic Brain Injury.

Based on accumulating evidence, patients recovering from mild and moderate traumatic brain injury (TBI) often experience increased sensitivity to stressful events. However, few studies have assessed on the effects and pathophysiological mechanisms of stress on TBI. In the current study, using a mouse model of moderate TBI, we investigated whether restraint stress (RS) regulates secondary neurodegeneration and neuronal cell death, which are commonly associated with neurological dysfunctions. Our data showed that RS significantly reduced body weight recovery, delayed the recovery of neurological functions (motor function, cognitive function and anxiety-like behavior) and exacerbated the brain lesion volume after moderate TBI. Immunofluorescence results indicated that moderate TBI-induced cell insults and blood-brain barrier leakage were aggravated by RS. Further Western blotting experiments showed that RS activated endoplasmic reticulum (ER) stress excessively after moderate TBI and decreased the number of NeuN-positive cells, but increased the number of CHOP/NeuN-co-positive cells by performing immunostaining in the injured cortex after moderate TBI. Moreover, RS increased the ratios of CHOP/Aß and CHOP/p-Tau co-positive cells in the injured cortex after moderate TBI. However, blocking ER stress with the classic ER stress inhibitor salubrinal remarkably decreased apoptosis and the levels of autophagy-related proteins in the mouse model of moderate TBI plus RS. Collectively, RS delays the recovery of neurological function and deteriorates morphological damage by excessively activating ER stress-mediated neurodegeneration, apoptosis and autophagy after moderate TBI. Thus, monitoring stress levels in patients recovering from non-severe TBI may merit consideration in the future.

Ruxolitinib exerts neuroprotection via repressing ferroptosis in a mouse model of traumatic brain injury.

Traumatic brain injury (TBI) is a major cause of death and disability worldwide. Various forms of cells death are involved in the pathological process of TBI, without exception to ferroptosis, which is mainly triggered by iron-dependent lipid peroxidation. Although there have been studies on ferroptosis and TBI, the effect of ruxolitinib (Ruxo), one type of FDA approved drugs for treating myelofibrosis, on the process of ferroptosis post-TBI is remained non-elucidated. Therefore, using a controlled cortical impact device to establish the mouse TBI model, we examined the effect of Ruxo on TBI-induced ferroptosis, in which the inhibitor of ferroptosis, Ferrostatin-1 (Fer-1) was used as a positive control. Moreover, we also respectively explored the effects of these two interventions on neurological deficits caused by TBI. We firstly examined the expression patterns of ferroptosis-related markers at protein level at different time points after TBI. And based on the expression changes of these markers, we chose 12 h post-TBI to prove the effect of Ruxo on ferroptosis. Importantly, we found the intensely inhibitory effect of Ruxo on ferroptosis, which is in parallel with the results obtained after Fer-1-treatment. In addition, these two treatments both alleviated the content of brain water and degree of neurodegeneration in the acute phase of TBI. Finally, we further confirmed the neuroprotective effect of Ruxo or Fer-1 via the wire-grip test, Morris water maze and open field test, respectively. Thereafter, the lesion volume and iron deposition were also measured to certificate their effects on the long-term outcomes of TBI. Our results ultimately demonstrate that inhibiting ferroptosis exerts neuroprotection, and this is another neuroprotective mechanism of Ruxo on TBI.

Autophagy Activation Represses Pyroptosis through the IL-13 and JAK1/STAT1 Pathways in a Mouse Model of Moderate Traumatic Brain Injury.

The newly highlighted research into programmed cell death (PCD), autophagy dependent cell death and pyroptotic cell death, has shown that these processes are both strongly correlated with the pathological progression of traumatic brain injury (TBI). However, their cross-talk in TBI remains unclear. Here, a moderate TBI model was established to explore the relationship between autophagy and pyroptosis. Rapamycin was used to activate the process of autophagy, which was impaired in the moderate TBI model, and this treatment reversed the expression of pyroptosis associated proteins, interleukin-13 (IL-13) and the pJAK-1 pathway, which were upregulated significantly after TBI. The level of IL-13 was downregulated, and the JAK-1 pathway was blocked to reveal the molecular mechanisms by which autophagy inhibits pyroptosis; these two treatments reduced the expression levels of pyroptosis associated proteins. In addition, these three interventions reduced the formation of neuronal NLRP3, the extent of brain edema, and the degree of cortical neuron degeneration. Furthermore, the deficit in motor function post-TBI was also markedly alleviated. Collectively, our results demonstrated that autophagy activation exerts a neuroprotective effect by inhibiting pyroptotic cell death in the moderate TBI model, and the inhibitory effect was dependent on the downregulation of IL-13 and repression of the JAK-1-STAT-1 signaling pathway.

Inhibiting ER Stress Weakens Neuronal Pyroptosis in a Mouse Acute Hemorrhagic Stroke Model.

Intracerebral hemorrhage (ICH) is a form of stroke, characterized by high morbidity and mortality and currently lacks specific therapy. ICH leads to endoplasmic reticulum (ER) stress, which can induce neurological impairment through crosstalk with programmed cell death (PCD). Pyroptosis, a newly discovered form of PCD, has received attention because of its close relationship with some certain diseases, such as traumatic brain injury and ischemic and hemorrhagic stroke. However, the relationship between ER stress and pyroptosis in ICH remains unclear. In this study, we investigated the role of ER stress in evoking neuronal pyroptosis and related mechanisms in a mouse ICH model. We used tauroursodeoxycholic acid (TUDCA) to inhibit ER stress and observed that TUDCA reduces neuronal pyroptosis and has a neuroprotective role. We explored the potential mechanisms underlying the regulation of neuronal pyroptosis by ER stress through testing the expression of interleukin-13 (IL-13). We found that ER stress inhibition alleviates neuronal pyroptosis through decreasing the expression of IL-13 after ICH. In summary, this study revealed that IL-13 is involved in ER stress-induced neuronal pyroptosis after ICH, pointing to IL-13 as a novel therapeutic target for ICH treatment.

High Capacity of Nutrient Accumulation by Invasive Solidago canadensis in a Coastal Grassland.

BACKGROUND: Solidago canadensis is a notorious invasive species from North America that is spreading across East China. It is invading some coastal grasslands and replacing native grass species. The effects of the S. canadensis invasion on soil nutrient cycling in the grasslands remain unclear. This study examined the effects of the invasion of S. canadensis on macronutrient accumulation in species aboveground part and soil. METHODS: Aboveground biomass, macronutrient (N, P, and K) pools in biomass, litter mass and decomposition rates, soil macronutrient availability and soil microbial biomass and enzyme activity that were related to nutrient transformation were compared between plots invaded by S. canadensis and uninvaded plots dominated by three different native grass species: Phacelurus latifolius, Phragmites australis, and Imperata cylindrica. RESULTS: S. canadensis had higher aboveground biomass, higher leaf N, P, and K concentrations, and consequently, a larger macronutrient pool size in the standing biomass. S. canadensis also produced more litter with higher N, P, and K concentrations and faster decomposition rates. The S. canadensis invasion did not change the total N, P, and K concentration in the topsoil (0-10 cm), but the invasion did increase their availability. The S. canadensis invasion did not increase the total soil organic matter (TSOM) content but did increase the soil microbial biomass and the activities of urease, alkaline phosphatase, invertase, amylase, and glucosidase in the topsoil. CONCLUSION: The invasion of S. canadensis accelerates the macronutrient cycling rate via increases in aboveground productivity and nutrient accumulation in standing biomass, faster nutrient release from litter and higher soil microbial activity. An enhanced nutrient cycling rate may further enhance its invasiveness through a positive feedback on soil processes.

Synthesis of Molecularly Imprinted Cryogels to Deplete Abundant Proteins from Bovine Serum.

Molecularly imprinted polyacrylamide cryogels were synthesized with pending templates (bovine serums of different concentrations). As the serum concentrations increased in the monomer solutions, the resulting cryogels could adsorb and deplete more proteins from serum samples. Due to the addition of vinyltriethoxysilane (VTEOS) in the prepolymerizing solutions, the polymers came as organic⁻inorganic hybrid materials. It endued the silica-modified amphoteric polyacrylamide cryogels with improved mechanical strengths. Scanning electron micrography (SEM), Infrared (IR) spectrometry, thermogravimetry-differential thermal analysis (TG-DTA), and X-ray photoelectron spectroscopy (XPS) were carried out to characterize these macroporous polymers. Amphoteric cryogels proved to be favorable materials recognizing and binding proteins. When used as liquid chromatography stationary phases, they were capable of simultaneously adsorbing various serum proteins. Electrophoresis showed that abundant proteins were gradually depleted by the cryogels prepared from increased ratios of bovine serums in the monomer solutions. As abundant proteins are always imprinted first, this sample per se imprinting method provides an effective and convenient way to deplete abundant proteins from complex samples such as serums, meanwhile concentrating and collecting scarce species therein.

Ice Squeezing Induced Multicolor Fluorescence Emissions from Polyacrylamide Cryogels.

Being short of conventional chromophores, polyacrylamide is generally not regarded as a fluorescent material. Exactly the polymerization of dilute solutions of acrylamide and N,N'-methylenebisacrylamide led to thick liquids at 60 °C, showing no fluorescence. Things changed when the phase transition of water was involved. The squeezing effect of ice crystals not only created polymeric solids (cryogels) at - 20 °C, but also endowed them unexpected fluorescence emissions. The macroporous cryogels are mainly blue fluorescent polymers. However yellow and red fluorescence were also achieved by changing the ingredient ratios. A series of instrumental detections revealed that the multicolor fluorescence were based on exquisite amido stacking induced from ice squeezing. If people make good use of the squeezing effect of the heaven-sent molecule to manipulate the interactions of monomer functionalities, cryogenic polymerization can be a promising method to produce diverse polymeric materials.

[Systematic Evaluation and Meta-analysis on Acupuncture for Peptic Ulcer].

OBJECTIVE: To systematically evaluate the clinical efficacy of acupuncture for peptic ulcer. METHODS: Randomized controlled trials of acupuncture for peptic ulcer were searched from China National Knowledge Infrastructure (CNKI), WanFang Database, Chinese Scientific and Technological Journals (VIP), China Biomedicine (CBM), PubMed and the Cochrane Library from the establishment time of databases to September, 2016. Data extraction and quality evaluation were implemented for the literature which met the inclusive criteria. The RevMan 5.3 software was used to make Meta-analysis. RESULTS: Sixteen papers including 1 570 patients of peptic ulcer were included. The results of Meta-analysis showed that there was no statistical significance between acupuncture and western medicine in the effective rate, the healing rate of ulcer area and the HP negative rate (all P>0.05); the recurrence rate of acupuncture was significantly lower than that of western medicine[RR=0.35, 95%CI (0.14, 0.84), P<0.05]. Acupuncture plus western medicine was significantly different from simple western medicine in the effective rate, the healing rate of ulcer area and the recurrence rate[RR=1.20,95% CI (1.04, 1.38), P=0.01; RR=1.29, 95% CI (1.06, 1.58), P=0.01;RR=0.27, 95% CI (0.16, 0.45), P<0.00001]. The analysis of evidence grade (GRADE) pre-sented that the healing rate of ulcer area and the HP negative rate of acupuncture were "low grade", and others were "extremely low grade". CONCLUSIONS: Acupuncture combined with western medicine has some advantages for peptic ulcer compared with the conventional western medicine, which needs further confirmation due to the lower evidence grade. Larger samples, randomized controlled trials with high quality are highly recommended.

Depletion of abundant human serum proteins by per se imprinted cryogels based on sample heterogeneity.

Macroporous cryogels were prepared and used to deplete abundant proteins. It was accomplished based on the sample heterogeneity rather than any exogenous assistance. Human serum was added in monomer solutions to synthesize molecularly imprinted polymers; therein some abundant proteins were imprinted in the polyacrylamide cryogels. Meanwhile the rare components remained aqueous. Chromatography and electrophoresis showed that albumin, serotransferrin, and most globulins were depleted by columns packed with the molecularly imprinted polymers. After the depletion, lower abundance proteins were revealed by SDS-PAGE, peptide fingerprint analysis, and identified by MALDI-TOF-MS. This is an example that a "per se imprint" protocol enables to gradually dimidiate proteomes, simplify sample complexities, and facilitate further proteome profiling or biomarker discovery.

[Effects of Direct Moxibustion of "Ganshu"(BL 18) on the Contents of T cells in Peripheral Blood in Rats with Precancerous Lesion of Primary Hepatocellular Carcinoma].

OBJECTIVE: To observe the effect of moxibustion stimulation of "Ganshu"(BL 18) region on contents of T cells in the peripheral blood in rats with Diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC), so as to explore its effective in improving immunoregulatory function. METHODS: Seventy male Wistar rats were randomly divided into control group (n=10), model group(n=15), direct moxibustion-15 s group(n=15), direct moxibustion-30 s group (n=15) and ginger-separated moxibustion group(n=15). The primary HCC precancerous lesion model was established by intraperitoneal injection of DEN (50 mg/kg), once every 3 days for 10 weeks. Moxibustion was applied to bilateral "Ganshu"(BL 18) for about 15 min (3 moxa-cones), or 30 min (six moxa-cones), with or without ginger-slice separation, once every other day for 10 consecutive weeks. The contents of T cells of CD 3+, CD 4+, CD 8+ and CD 4+/CD 8+ ratio in the peripheral blood were detected with Flow Cytometey(FCM), pathological changes of liver were observed by light microscope after hematoxylin-eosin(HE) stain. RESULTS: Compared to the control group, the contents of blood CD 3+ and CD 4+ T cells and ratio of CD 4+/CD 8+ were significantly down-regulated, while that of CD 8+T cells was obviously increased in the model group(P<0.05,P<0.01). After moxibustion intervention, the decreased CD 3+ and CD 4+T cells and CD 4+/CD 8+ levels and the increased CD 8+T cell contents were reversed in all the 3 moxibustion groups (P<0.05, P<0.01), except CD 3+ in the ginger-separated moxibustion group (P>0.05). There were no significant differences among the three moxibustion groups in the CD 3+, CD 4+, CD 8+ and CD 4+/CD 8+ levels (P>0.05). In addition, the pathological changes of liver tissue as central vein deviation or absence, disordered arrangement of hepatic cords, narrowing of the hepatic sinusoid, hyperplasia of collagen fibers, formation of tuberosis, unevenness of liver cells with nuclear anachromasis and higher heteromorphism, and macronucleus oncocytes in primary HCC rats were not observed or milder after moxibustion intervention. CONCLUSIONS: Direct moxibustion and ginger-separated moxibustion can improve pathological changes of hepatic cells in rats with HCC, which may be associated with its actions in raising blood CD 3+ and CD 4+ T cell contents and reducing CD 8+ T levels to improve immune function.

[Effects of acupuncture stimulation of different acupoint groups on sleeping latency, serum and hippocampal TNF-α and IL-25 contents in rats with gastric mucosal injury].

OBJECTIVE: To observe the effect of acupuncture intervention on gastric ulcer (GU) and sleeping quality from the viewpoint of brain-gut axis which plays an important role in the regulation of many vital functions in the body. METHODS: Forty male Wistar rats were randomized into normal control, GU model, acupuncture of "Zhongwan"(CV 12)-"Zusanli"(ST 36, gastric function regulating acupoints), acupuncture of "Shenmai" (BL 62)-"Zhaohai" (KI 6, sleep-promotion acupoints), and acupuncture of CV 12-ST 36+ BL 62-KI 6 (combined treatment) groups, with 8 rats in each group. GU model was established by intragastric perfusion of dehydrated alcohol (1 mL/rat), and sleep model established by intraperitoneal injection of pentobarbital sodium (40 mg/kg) after the last treatment. The abovementioned acupoints were punctured with filiform needles and stimulated by manipulating the needle for about 30 s, once every 5 min during 20 min of needle retention. The treatment was conducted once daily for five days. The contents of tumor necrosis factor-alpha (TNF-α) and interleukin-25(IL-25) in the serum and hippocampal tissues were detected by ELISA. RESULTS: Compared with the normal control group, the gastric ulcer index score, barbiturate-induced sleeping time, and TNF-α and IL-25 contents in both serum and hippocampus were significantly increased in the model group (P < 0.01). Following acupuncture treatment, in comparison with the model group, the gastric ulcer index score, barbiturate-induced sleeping time, and TNF-α and IL-25 contents in both serum and hippocampus were significantly down-regulated in the CV 12-ST 36, BL 62-KI 6 and combined treatment groups (P < 0.01, P < 0.05). The effects of the CV 12-ST 36 and combined treatment groups were remarkably superior to those of the BL 62-KI 6 group in down-regulating ulcer index score, serum IL-25, and hippocampal TNF-α and IL-25 contents (P < 0.01, P < 0.05). In addition, the effects of the BL 62-KI 6 and combined treatment groups was considerably better than that of the CV 12-ST 36 group in shortening barbiturate-induced sleeping time (P < 0.01, P < 0.05). The effect of the combined treatment group was markedly better than that of the CV 12-ST 36 and BL 62-KI 6 groups in lowering serum TNF-α content (P < 0.05). CONCLUSION: Acupuncture stimulation of CV 12, ST 36, KI 6 and BL 62 can relieve the gastric mucosal lesion, and shorten barbiturate-induced sleeping time in gastric ulcer rats, which may be related to its effects in reducing TNF-α and IL-25 contents in the serum and hippocampus tissues, suggesting a correlation between the gastrointestinal disorder and sleeping.