Identification of tumor-suppressive miRNAs that target amino acid transporter LAT1 and exhibit anti-proliferative effects on cholangiocarcinoma cells.

Amino acid transporter LAT1 is highly upregulated in various cancer types, including cholangiocarcinoma (CHOL), and contributes to the rapid proliferation of cancer cells and disease progression. However, the molecular mechanisms underlying the pathological upregulation of LAT1 remain largely unknown. This study pursued the possibility of miRNA-mediated regulation of the LAT1 expression in CHOL cells. Using online target prediction methods, we extracted five candidate miRNAs commonly predicted to regulate the LAT1 expression. Three of them, miR-194-5p, miR-122-5p, and miR-126-3p, were significantly downregulated in CHOL cancer compared to normal tissues. Correlation analysis revealed weak-to-moderate negative correlations between the expression of these miRNAs and LAT1 mRNA in CHOL cancer tissues. We selected miR-194-5p and miR-122-5p for further analyses and found that both miRNAs functionally target 3'UTR of LAT1 mRNA by a luciferase-based reporter assay. Transfection of the miRNA mimics significantly suppressed the LAT1 expression at mRNA and protein levels and inhibited the proliferation of CHOL cells, with a trend of affecting intracellular amino acids and amino acid-related signaling pathways. This study indicates that the decreased expression of these LAT1-targeting tumor-suppressive miRNAs contributes to the upregulation of LAT1 and the proliferation of CHOL cells, highlighting their potential for developing novel cancer therapeutics and diagnostics.

Mapping multi-seasonal habitats of giant pandas to identify seasonal shifts.

As a flagship species of biodiversity conservation globally, the giant panda has seasonal migration to cope with seasonal changes in available resources. Here, we have mapped the spatial distribution of multi-seasonal habitats of the giant panda across the Baishuijiang reserve in China. Results show that the spatial patterns are different in different seasons, generally, large patches are observed in the western part, while staggered clusters occur in the middle and eastern parts. That is, suitable habitats for giant pandas are mostly distributed in the west part. More than 75% of the predicted suitable habitats are within the core zone of the reserve year-round, indicating the core zone essentially meet giant panda's ecological needs, although this range could potentially be expanded. This study provides valuable insights into the spatiotemporal migration patterns of endangered species and helps to guide conservation planning.

An Improved Model of Single-Event Transients Based on Effective Space Charge for Metal-Oxide-Semiconductor Field-Effect Transistor.

In this paper, a single-event transient model based on the effective space charge for MOSFETs is proposed. The physical process of deposited and moving charges is analyzed in detail. The influence of deposited charges on the electric field in the depletion region is investigated. The electric field decreases in a short time period due to the neutralization of the space charge. After that, the electric field increases first and then decreases when the deposited charge is moved out. The movement of the deposited charge in the body mainly occurs through ambipolar diffusion because of its high-density electrons and holes. The derivation of the variation in electric field in the depletion region is modeled in the physical process according to the analysis. In combination with the ambipolar diffusion model of excessive charge in the body, a physics-based model is built to describe the current pulse in the drain terminal. The proposed model takes into account the influence of multiple factors, like linear-energy transfer (LET), drain bias, and the doping concentration of the well. The model results are validated with the simulation results from TCAD. Through calculation, the root-mean-square error (RMSE) between the simulation and model is less than 3.7 × 10-4, which means that the model matches well with the TCAD results. Moreover, a CMOS inverter is simulated using TCAD and SPICE to validate the applicability of the proposed model in a circuit-level simulation. The proposed model captures the variation in net voltage in the inverter. The simulation result obviously shows the current plateau effect, while the relative error of the pulse width is 23.5%, much better than that in the classic model. In comparison with the classic model, the proposed model provides an RMSE of 7.59 × 10-5 for the output current curve and an RMSE of 0.158 for the output voltage curve, which are significantly better than those of the classic model. In the meantime, the proposed model does not produce extra simulation time compared with the classic double exponential model. So, the model has potential for application to flow estimation of the soft error rate (SER) at the circuit level to improve the accuracy of the results.

A High-Reliability 12T SRAM Radiation-Hardened Cell for Aerospace Applications.

The static random-access memory (SRAM) cells used in the high radiation environment of aerospace have become highly vulnerable to single-event effects (SEE). Therefore, a 12T SRAM-hardened circuit (RHB-12T cell) for the soft error recovery is proposed using the radiation hardening design (RHBD) concept. To verify the performance of the RHB-12T, the proposed cell is simulated by the 28 nm CMOS process and compared with other hardened cells (Quatro-10T, WE-Quatro-12T, RHM-12T, RHD-12T, and RSP-14T). The simulation results show that the RHB-12T cell can recover not only from single-event upset caused by their sensitive nodes but also from single-event multi-node upset caused by their storage node pairs. The proposed cell exhibits 1.14×/1.23×/1.06× shorter read delay than Quatro-10T/WE-Quatro-12T/RSP-14T and 1.31×/1.11×/1.18×/1.37× shorter write delay than WE-Quatro-12T/RHM-12T/RHD-12T/RSP-14T. It also shows 1.35×/1.11×/1.04× higher read stability than Quatro-10T/RHM-12T/RHD-12T and 1.12×/1.04×/1.09× higher write ability than RHM-12T/RHD-12T/RSP-14T. All these improvements are achieved at the cost of a slightly larger area and power consumption.

Combination effects of amino acid transporter LAT1 inhibitor nanvuranlat and cytotoxic anticancer drug gemcitabine on pancreatic and biliary tract cancer cells.

BACKGROUND: Cytotoxic anticancer drugs widely used in cancer chemotherapy have some limitations, such as the development of side effects and drug resistance. Furthermore, monotherapy is often less effective against heterogeneous cancer tissues. Combination therapies of cytotoxic anticancer drugs with molecularly targeted drugs have been pursued to solve such fundamental problems. Nanvuranlat (JPH203 or KYT-0353), an inhibitor for L-type amino acid transporter 1 (LAT1; SLC7A5), has novel mechanisms of action to suppress the cancer cell proliferation and tumor growth by inhibiting the transport of large neutral amino acids into cancer cells. This study investigated the potential of the combined use of nanvuranlat and cytotoxic anticancer drugs. METHODS: The combination effects of cytotoxic anticancer drugs and nanvuranlat on cell growth were examined by a water-soluble tetrazolium salt assay in two-dimensional cultures of pancreatic and biliary tract cancer cell lines. To elucidate the pharmacological mechanisms underlying the combination of gemcitabine and nanvuranlat, we investigated apoptotic cell death and cell cycle by flow cytometry. The phosphorylation levels of amino acid-related signaling pathways were analyzed by Western blot. Furthermore, growth inhibition was examined in cancer cell spheroids. RESULTS: All the tested seven types of cytotoxic anticancer drugs combined with nanvuranlat significantly inhibited the cell growth of pancreatic cancer MIA PaCa-2 cells compared to their single treatment. Among them, the combined effects of gemcitabine and nanvuranlat were relatively high and confirmed in multiple pancreatic and biliary tract cell lines in two-dimensional cultures. The growth inhibitory effects were suggested to be additive but not synergistic under the tested conditions. Gemcitabine generally induced cell cycle arrest at the S phase and apoptotic cell death, while nanvuranlat induced cell cycle arrest at the G0/G1 phase and affected amino acid-related mTORC1 and GAAC signaling pathways. In combination, each anticancer drug basically exerted its own pharmacological activities, although gemcitabine more strongly influenced the cell cycle than nanvuranlat. The combination effects of growth inhibition were also verified in cancer cell spheroids. CONCLUSIONS: Our study demonstrates the potential of first-in-class LAT1 inhibitor nanvuranlat as a concomitant drug with cytotoxic anticancer drugs, especially gemcitabine, on pancreatic and biliary tract cancers.

Homeodomain-interacting protein kinase 2 regulates NLRP3 inflammasome activation through endoplasmic reticulum stress in septic liver injury.

OBJECTIVE: Septic liver injury is a major burden for the clinical management of sepsis. Hepatocyte cell death plays a crucial pathophysiological role in sepsis. A recent study proposed that NLRP3 inflammasome-mediated pyroptosis participates in septic liver injury. Therefore, investigating the mechanism controlling this process may help manage sepsis. METHODS: We investigated the role of homeodomain-interacting protein kinase 2 (HIPK2) in regulating the NLRP3 inflammasome in vivo using mouse models and in vitro in primary hepatocytes. RESULTS: HIPK2 could improve liver injury and survival in a mouse model of sepsis. Overexpression of HIPK2 could suppress NLRP3 and caspase-1-p20 expression, while HIPK2 knockdown led to higher levels of these two molecules. Importantly, HIPK2 could suppress endoplasmic reticulum (ER) stress. Pharmacologically inhibiting ER stress could abolish activation of the NLRP3 inflammasome in hepatocytes with HIPK2 knockdown. CONCLUSION: HIPK2 can regulate ER stress and NLRP3 inflammasome activation in the liver during sepsis, and HIPK2-mediated suppression of ER stress participates in regulating NLRP3 inflammasome activation. The present study highlights the role of HIPK2 in regulating the inflammasome in septic liver injury, which may serve as a target for managing sepsis.

Critical role of caveolin-1 in intestinal ischemia reperfusion by inhibiting protein kinase C ßII.

Intestinal ischemia reperfusion (I/R) is a common clinical pathological process. We previously reported that pharmacological inhibition of protein kinase C (PKC) ßII with a specific inhibitor attenuated gut I/R injury. However, the endogenous regulatory mechanism of PKCßII inactivation is still unclear. Here, we explored the critical role of caveolin-1 (Cav1) in protecting against intestinal I/R injury by regulating PKCßII inactivation. PKCßII translocated to caveolae and bound with Cav1 after intestinal I/R. Cav1 was highly expressed in the intestine of mice with I/R and IEC-6 cells stimulated with hypoxia/reoxygenation (H/R). Cav1-knockout (KO) mice suffered from worse intestinal injury after I/R than wild-type (WT) mice and showed extremely low survival due to exacerbated systemic inflammatory response syndrome (SIRS) and remote organ (lung and liver) injury. Cav1 deficiency resulted in excessive PKCßII activation and increased oxidative stress and apoptosis after intestinal I/R. Full-length Cav1 scaffolding domain peptide (CSP) suppressed excessive PKCßII activation and protected the gut against oxidative stress and apoptosis due to I/R injury. In summary, Cav1 could regulate PKCßII endogenous inactivation to alleviate intestinal I/R injury. This finding may represent a novel therapeutic strategy for the prevention and treatment of intestinal I/R injury.

[Double-channel expansion of the subcutaneous lower abdominal tunnel for sequential inguinal lymph node dissection in penile cancer - Report of six cases].

OBJECTIVE: To investigate the safety and efficacy of the two-channel dilatation procedure for subcutaneous tunneling in the lower abdomen during pelvic lymph node dissection for penile cancer. METHODS: A retrospective analysis was conducted on the clinical data of 6 patients treated from January 2020 to December 2022 using the dual-channel expansion technique for penile cancer lymph node dissection. RESULTS: All 6 cases ( 12 sides) successfully underwent prophylactic inguinal lymph node dissection. The average laparoscopic dissection time was ( 82.50 ± 12.08) minutes per side, with an average blood loss of （28.33 ± 10.95） ml. The number of lymph nodes dissected was （11.16 ± 1.02） for the superficial group and ( 0.67 ± 0.74 ) for the deep group. Postoperative pathology was negative in all cases. The average postoperative hospital stay was （7.33 ± 1.60 ） days, with a catheter removal time of （12.00 ± 2.06）days. Postoperative complications included abnormal skin sensations in 5 sides, lower limb edema in 3 sides, lymphedema in 3 sides, and cellulitis in 1 side. During a follow-up period of （20.60 ± 12.51）months, there were no instances of tumor recurrence or metastasis in the inguinal region among the patients. CONCLUSION: The dual-channel expansion technique for inguinal lymph node dissection via a subcutaneous tunnel is a safe and feasible treatment for penile cancer. It has a low complication rate, allows for thorough dissection of inguinal lymph nodes, and offers advantages in terms of surgical time.

Logical Resolving-Based Methodology for Efficient Reliability Analysis.

With the CMOS technology downscaling to the deep nanoscale, the aging effects of devices degrade circuit performance and even lead to functional failure. The stress analysis is critical to evaluate the influence of aging effects on digital circuits. Some related analytical work has recently focused on reliability-aware circuit analysis. Nevertheless, the aging dependence among different devices is not considered, which will induce errors of degradation evaluation in the digital circuit. In order to improve the accuracy of reliability-aware static timing analysis, an improved analytical method is proposed by employing logical resolving. Experimental results show that the proposed method has a better evaluation accuracy of aging path delay than traditional strategies. For aging timing evaluation on aging paths, excessive pessimism can be reduced by employing the proposed method. And, a 378× speedup is achieved while having a 0.56% relative error compared with precise SPICE simulation. Moreover, the circuit performance sacrifice of an aging-aware synthesis flow with the proposed method can be decreased. Due to the high efficiency and high accuracy, the proposed method can meet the speed demands of large-scale digital circuit reliability analysis while achieving transistor simulation accuracy.

[Application of groin flap combined with medial plantar artery perforator flap for degree â ¢- â £ defects of multiple fingers].

Objective: To investigate the effectiveness of groin flap combined with medial plantar artery perforator flap (MPAP) for degree Ⅲ-Ⅳ defects of multiple fingers. Methods: Between January 2018 and June 2019, 12 patients with degree Ⅲ-Ⅳ defects of multiple fingers caused by crushing were admitted. There were 9 males and 3 females with a median age of 29 years (range, 16-42 years). The mean interval between the injury and admission was 3 hours (range, 1-9 hours). The injured fingers of 7 cases were index and middle fingers, 4 cases were middle and ring fingers, and 1 case was index, middle, and ring fingers. All fingers were taken thorough debridement and covered by the vacuum sealing drainage device during the emergency operation. The mean interval between the debridement and flap repairing was 18 hours (range, 12-36 hours). During the first-stage operation, the iliac bone graft was used to reconstruct bone frame, and the proximal interphalangeal (PIP) joint from the foot was transferred as the digital PIP joint, then the thin groin flap and MPAP were tailored to cover the dorsal and palmar defects, respectively. The size of the groin flap was 7.0 cm×4.5 cm-14.0 cm×9.0 cm, and the size of the MPAP was 8.0 cm×4.5 cm-14.0 cm×6.5 cm. The abdominal donor site was directly sutured, and the foot was repaired with full-thickness skin grafting. The flaps were separated into the finger shape at the second-stage. Results: All the flaps survived, and the wounds healed by first intention; the incisions in the donor site healed by first intention, and the skin grafts survived completely. All patients were followed up 12-18 months (mean, 16 months). At last follow-up, the injured finger was similar to the contralateral one in terms of texture, appearance, and color. The mean two-point discrimination was 8 mm (range, 6-10 mm), and the sensate level recovered to the S 3-S 4. According to the Michigan Hand Outcomes Questionnaire (MHQ), the reconstructed hand function was excellent in 8 cases and good in 4 cases. There was no complication in the donor sites. Conclusion: The degree Ⅲ-Ⅳ defects of multiple fingers were repaired by the groin flap and MPAP, and the reconstructed fingers can perform good texture and motion with being sensate, with less sacrifice on the foot.

Bioinformatics analysis reveals the clinical significance of GIPC2/GPD1L for colorectal cancer using TCGA database.

Background: Colorectal cancer (CRC) causes 700,000 deaths annually and is the fourth deadliest cancer in the world after lung, liver, and stomach cancer. Since CRC is difficult to detect early and has a poor prognosis, it is critical to develop novel biomarkers for its diagnosis, prognosis, and treatment. Methods: The GIPC2 expression in colorectal cancer was examined by the TCGA database analysis, IHC from the human protein atlas and qRT-PCR tests. GO and KEGG enrichment analyses were performed for genes that were both correlated with the expression of GIPC2 and GPD1L. The receiver operating characteristic curve (ROC) analysis and Kaplan-Meier (KM) survival analysis were applied to analyze the prognostic value of GIPC2 and GPD1L for overall survival (OS) and progress free interval (PFI) of CRC patients. Results: We found that GIPC2 was low expressed in colorectal cancer and highly related with the CRC clinical-stage grade and TNM stage. Furthermore, GPD1L is correlated with GIPC2 via the correlation analysis in CRC and they were associated with several important cancer-related pathways. GIPC2 and GPD1L exhibited good diagnostic and prognostic predictive ability for patients with CRC. Conclusions: These results revealed new biomarkers in CRC, we proposed that the GIPC2/GPDL1 might be potential diagnostic and prognostic indicators for CRC, which provides a theoretical basis for our subsequent cellular and animal experiments, so as to reveal the occurrence and development mechanism of CRC more comprehensively.

Systemic Immune-Inflammation Index (SII) Can Be an Early Indicator for Predicting the Severity of Acute Pancreatitis: A Retrospective Study.

OBJECTIVE: Systemic immune-inflammation index (SII) is a new systemic inflammatory prognostic indicator associated with outcomes in patients with different tumors. Studies have shown an association between SII and many chronic/acute inflammatory diseases. This study aimed at exploring whether SII can be used as an effective parameter for predicting the severity of acute pancreatitis (AP). METHODS: A total of 101 acute pancreatitis patients were enrolled in this study (mild acute pancreatitis (MAP): n = 73 and severe acute pancreatitis (SAP): n = 28). Patient demographics and SII were analyzed using the chi-square test, Student's t-test, and Mann-Whitney U-test. A receiver operating characteristic curve was generated to test the potential of using neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), and SII to predict AP's severity. Logistic regression analysis was performed to determine major risk factors. RESULTS: Patients with SII value ≥2207.53 had a higher probability of having SAP (sensitivity = 92.9%, specificity = 87.7%, and AUC = 0.920), and SII was a significantly better predictive value than PLR and NLR. Logistic regression analysis results showed SII could differentiate MAP from SAP as a major risk factor. CONCLUSION: This study has shown that SII is a potential indicator for predicting the severity of acute pancreatitis. The findings suggested that SII is more sensitive and specific than NLR and PLR in predicting the severity of acute pancreatitis.

Logic-signal-based multiplex detection of MiRNAs with high tension hybridization and multiple signal amplification.

MicroRNAs (miRNAs) are crucial regulators of gene expression. The abnormal expression of miRNA is often closely related to many diseases. However, the accurate clinical profiling of miRNA expression remains a great challenge due to the high similarity and wide variety of miRNA sequence structures. Here, we report a highly specific and sensitive multiplex miRNA detection scheme with high tension hybridization and dual signal amplification based on the recyclable autocatalytic DNAzyme and a light harvesting conjugated polymer. Multiple signals can be read out simultaneously by single excitation through the efficient multiple fluorescence resonance energy transfer (FRET) between the conjugated polymer and different small molecule dyes. In addition, different types of logic gates can also be operated by observing the emission intensities of the labeling dyes with miRNAs as inputs, thus giving rise to a new way for the specific detection of certain miRNAs according to the logic signals. Furthermore, we successfully applied the strategy for multiple miRNA detection in cell lysates and the results agree well with those of qRT-PCR. Thus, we believe that this platform holds great potential for miRNA detection in biological samples.

Metabolomics-Based Discovery of Molecular Signatures for Triple Negative Breast Cancer in Asian Female Population.

Triple negative breast cancer (TNBC) is a devastating cancer disease characterized by its poor prognosis, distinct metastatic patterns, and aggressive biological behavior. Research indicates that the prevalence and presentation of TNBC varies among races, with Asian TNBC patients more commonly presenting with large invasive tumors, high node positivity, and high histologic grade. In this work, we applied ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS)-based metabolomics to discover metabolic signatures in Asian female TNBC patients. Serum samples from 31 TNBC patients and 31 healthy controls (CN) were involved in this study. A total of 2860 metabolic features were detected in the serum samples. Among them, 77 metabolites, whose levels were significantly different between TNBC with CN, were confirmed. Using multivariate statistical analysis, literature mining, metabolic network and pathway analysis, we performed an in-depth study of the metabolic alterations in the Asian TNBC population. In addition, we discovered a panel of metabolic signatures that are highly correlated with the 5-year survival rate of the TNBC patients. This metabolomic study provides a better understanding of the metabolic details of TNBC in the Asian population.

Tofu intake is inversely associated with risk of breast cancer: A meta-analysis of observational studies.

Observational studies on the association between tofu intake and breast cancer incidence have reported inconsistent results. We reviewed the current evidence and quantitatively assessed this association by conducting a dose-response meta-analysis. The electronic databases PubMed and EMBASE were searched for relevant studies published up to August, 2018. We included epidemiological studies that reported relative risks (RRs) or odds ratios (ORs) with 95% confidence intervals (CIs) for the association between tofu intake and breast cancer risk. A total of 14 studies (2 cohort studies, 12 case-control studies) were included in the meta-analysis. The overall OR of breast cancer for highest vs lowest intake of tofu was 0.78 (95% CI 0.69-0.88), with moderate heterogeneity (P = 0.011, I2 = 49.7%). Dose-response analysis based on 5 case-control studies revealed that each 10 g/d increase in tofu intake was associated with 10% reduction in the risk of breast cancer (95% CI 7%-13%, P = 0.037, I2 = 40.8%). In summary, our findings suggest an inverse dose-response association between tofu intake and risk of breast cancer. However, owing to the limitations of case-control studies, more properly designed prospective studies are warranted to confirm this association.

Boron delivery for boron neutron capture therapy targeting a cancer-upregulated oligopeptide transporter.

Boron neutron capture therapy (BNCT) is a radiotherapy utilizing the neutron capture and nuclear fission reaction of 10B taken up into tumor cells. The most commonly used boron agent in BNCT, p-borono-l-phenylalanine (BPA), is accumulated in tumors by amino acid transporters upregulated in tumor cells. Here, by using dipeptides of BPA and tyrosine (BPA-Tyr and Tyr-BPA), we propose a novel strategy of selective boron delivery into tumor cells via oligopeptide transporter PEPT1 upregulated in various cancers. Kinetic analyses indicated that BPA-Tyr and Tyr-BPA are transported by oligopeptide transporters, PEPT1 and PEPT2. The intrinsic oligopeptide transport activity in tumor cells clearly correlated with PEPT1 protein expression level but not with PEPT2, suggesting that PEPT1 is the predominant oligopeptide transporter at least in tumor cell lines. Furthermore, using BPA-Tyr and Tyr-BPA, boron was successfully delivered into PEPT1-expressing pancreatic cancer AsPC-1 cells via a PEPT1-mediated mechanism. Intravenous administration of BPA-Tyr into the mice bearing AsPC-1 xenograft tumors resulted in significant boron accumulation in the tumors. It is proposed that the oligopeptide transporters, especially PEPT1, are promising candidates for molecular targets of boron delivery in BNCT. The BPA-containing dipeptides would have a potential for the development of novel boron carriers targeting PEPT1.

Evaluating giant panda as a surrogate species for conservation co-occurring species in the Baishuijiang National Nature Reserve.

The establishment of nature reserves is a key approach for biodiversity conservation worldwide. However, the effectiveness of nature reserves established by protecting the habitat needs of surrogate species is questioned. In this study, the Baishuijiang National Nature Reserve (Baishuijiang NNR), located in the Minshan Mountains, China, which is established mainly for the conservation of giant panda (a surrogate for the conservation of other endangered species) was selected. We quantitatively evaluated the conservation effectiveness of the reserve for giant panda and co-occurring species (here, seven protected species) using a maximum entropy model (Maxent), and analyzed spatial congruence between giant panda and other seven species. Results shown that the habitat of giant panda generally included the habitat of other seven protected species, suggesting that conservation of giant panda habitat also allows the conservation for the habitat of almost co-occurring species. Hence, the natural reserve established for giant panda as a surrogate species has a relatively high effectiveness. A high proportion of the suitable habitat for six species is inside the core zone, but a high proportion of the suitable habitat for two species is located in the experimental and buffer zones. Thus, the two species are affected by human activities. To improve the conservation effectiveness of the nature reserve, the management zones need to be amended. The result of the study will be beneficial for future conservation and management of the reserve. This study provides an effective method for evaluating the conservation effectiveness of nature reserves in other area of the worldwide.

Evaluation and efficacy of metal oxides in terms of antibacterial activity and toxic chemical degradation.

Inorganic metal oxides Ag2O, CuO and ZnO were examined using SEM, XRD, TGA and ICP spectroscopy to analyze their structures and physical properties in terms of resistance to germs and toxic chemicals. Zone of inhibition testing and the plate-counting method were used in this study to examine the antibacterial activities of the metal oxides against Gram-negative Escherichia coli (E. coli) and Pseudomonas aeruginosa (P. aeruginosa), and Gram-positive Staphylococcus aureus (S. aureus) and Bacillus subtilis (B. subtilis). Furthermore, 2­chloro­ethyl ethyl sulfide (2­CEES) was used to study the degradation efficacy of the metal oxides by the NMR method. The objective of the study was to develop and evaluate metal oxides that are able to protect against chemical and biological warfare agents. Excellent antibacterial and catalytic toxic chemical degradation properties were obtained.

Fish oil alleviates liver injury induced by intestinal ischemia/reperfusion via AMPK/SIRT-1/autophagy pathway.

AIM: To evaluate whether fish oil (FO) can protect liver injury induced by intestinal ischemia/reperfusion (I/R) via the AMPK/SIRT-1/autophagy pathway. METHODS: Ischemia in Wistar rats was induced by superior mesenteric artery occlusion for 60 min and reperfusion for 240 min. One milliliter per day of FO emulsion or normal saline was administered by intraperitoneal injection for 5 consecutive days to each animal. Animals were sacrificed at the end of reperfusion. Blood and tissue samples were collected for analyses. AMPK, SIRT-1, and Beclin-1 expression was determined in lipopolysaccharide (LPS)-stimulated HepG2 cells with or without FO emulsion treatment. RESULTS: Intestinal I/R induced significant liver morphological changes and increased serum alanine aminotransferase and aspartate aminotransferase levels. Expression of p-AMPK/AMPK, SIRT-1, and autophagy markers was decreased whereas tumor necrosis factor-α (TNF-α) and malonaldehyde (MDA) were increased. FO emulsion blocked the changes of the above indicators effectively. Besides, in LPS-stimulated HepG2 cells, small interfering RNA (siRNA) targeting AMPK impaired the FO induced increase of p-AMPK, SIRT-1, and Beclin-1 and decrease of TNF-α and MDA. SIRT-1 siRNA impaired the increase of SIRT-1 and Beclin-1 and the decrease of TNF-α and MDA. CONCLUSION: Our study indicates that FO may protect the liver against intestinal I/R induced injury through the AMPK/SIRT-1/autophagy pathway.

An H-NS Family Protein, Sfh, Regulates Acid Resistance by Inhibition of Glutamate Decarboxylase Expression in Shigella flexneri 2457T.

The glutamate-dependent acid-resistance system is the most effective acid tolerance pathway in Shigella, allowing survival in extremely acidic environments. However, the regulation of this system in Shigella remains elusive. In the current study, we identified significant differences in the levels of glutamate decarboxylase between three Shigella flexneri strains with different levels of acid resistance using blue native-polyacrylamide gel electrophoresis (PAGE) and isoelectric focusing (IEF)/sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The results showed that the degree of acid resistance and the levels of GadA/B were significantly lower in strain 2457T compared with two other S. flexneri strains. It has been reported that plasmid pSf-R27 is expressed in strain 2457T but not in the other 142 sequenced S. flexneri isolates. pSf-R27 encodes protein Sfh, which belongs to a family of histone-like nucleoid-structuring (H-NS) proteins that participate in the transcriptional control of glutamate-dependent acid resistance, implicating pSf-R27 in the lower acid resistance of strain 2457T. Transformation of pSf-R27 or sfh alone into strain 301 resulted in decreased expression of GadA/B in the recombinant strains. Thus, we confirmed that H-NS family protein Sfh, bound to the gadA/B regulatory region and regulates the expression of glutamate decarboxylase at the transcriptional level. We also examined the acid tolerance of the wild-type and recombinant strains using flow cytometry and determined that the acid tolerance of S. flexneri is closely related to the expression of GadA/B. These findings further our understanding of the acid tolerance of S. flexneri, especially via the glutamate-dependent pathway.