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The relative conservation of the influenza hemagglutinin (HA) stem compared to that of the immunodominant HA head makes the HA stem an attractive target for broadly protective influenza vaccines. Here we report the first-in-human, dose-escalation, open-label trial (NCT04579250) evaluating an unadjuvanted group 2 stabilized stem ferritin nanoparticle vaccine based on the H10 A/Jiangxi-Donghu/346/2013 influenza HA, H10ssF, in healthy adults. Participants received a single 20 mcg dose (n = 3) or two 60 mcg doses 16 weeks apart (n = 22). Vaccination with H10ssF was safe and well tolerated with only mild systemic and local reactogenicity reported. No serious adverse events occurred. Vaccination significantly increased homologous H10 HA stem binding and neutralizing antibodies at 2 weeks after both first and second vaccinations, and these responses remained above baseline at 40 weeks. Heterologous H3 and H7 binding antibodies also significantly increased after each vaccination and remained elevated throughout the study. These data indicate that the group 2 HA stem nanoparticle vaccine is safe and induces stem-directed binding and neutralizing antibodies., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Purpose: As a major structural component of the outer membrane of Gram-negative bacteria, lipopolysaccharide (LPS) has been detected in the blood circulation and tissues in patients with chronic diseases and cancers, which plays a critical role in the tumor formation and progression. However, the biological role of LPS in human intrahepatic cholangiocarcinoma remains unclear. The aims of this study were to investigate the role of LPS in the malignant progression of intrahepatic cholangiocarcinoma., Methods: The cell migration and invasion capacities of cholangiocarcinoma cell lines were evaluated by Boyden chamber assays. Expression levels of the key molecules involved in the PI3K/AKT signaling and METTL3 were detected by qPCR and western blot. The molecular mechanism by which LPS promotes the malignant behaviors was investigated by using siRNAs, plasmids and small molecule inhibitors., Results: In vitro experiments showed that exogenous LPS treatment promoted cell migration and invasion capacities in both QBC939 and HUCCT1 cell lines, while did not affect cell proliferation and apoptosis. Mechanistically, exogenous LPS treatment had been proved to induce the increased expression of METTL3 and activate the downstream PI3K/AKTsignaling pathway. In addition, suppression of METTL3 expression reduced cell proliferation, migration and invasion capacities in both cell lines. Furthermore, inhibition of METTL3 expression or inhibition of PI3K/AKT signaling decreased LPS-induced cell migration and invasion capacities. Moreover, knockdown of METTL3 or inhibition of METTL3 significantly inhibited LPS-induced activation of the PI3K/AKT signaling., Conclusion: In general , these results suggest that the LPS-METTL3-PI3K/AKT signal axis promotes cell migration and invasion in ICC, which contributes to a reduced overall survival in patients with ICC. It may broaden the horizon of cancer therapy with potential therapeutic targets., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Objectives: To investigate the effect of electroacupuncture(EA) at "Changbing Decoction" on alleviating ulcerative colitis (UC) and regulating the polarization of colonic macrophages in rats, so as to explore its mechanisms underlying improvement of UC., Methods: Twenty-six male SD rats were randomly divided into 4 groups：normal group(6 rats), model group(8 rats), EA group(6 rats), and western medication group(6 rats). The rat model of UC was established by using 5% dextran sulfate sodium (DSS) solution drinking water for 7 days, followed by drinking 1% DSS solution during treatment period. After 7-day model establishment, EA treatment(10 Hz/50 Hz, 20 min) was applied to "Zhongwan"(CV12), bilateral "Tianshu"(ST25) and "Shangjuxu"(ST37) for 3 d, and rats in the western medication group were given mesalazine suspension(200 mg/kg) by gavage for 3 d. The body weight, spleen weight and colon length of rats were measured. The disease activity index (DAI) score was evaluated. The morphological changes and inflammatory cell infiltration of colon were detected after HE staining and pathological scores were eva-luated. The contents of tumor necrosis factor α(TNF-α), interleukin(IL)-1β, IL-2 and IL-10 in serum were detected by ELISA. The protein expressions of M1 and M2 macrophage markers nitric oxide synthase (iNOS) and arginase 1(Arg1) were detected by fluorescence double staining and Western blot, respectively. Quantitative real-time PCR was used to detect iNOS and Arg1 mRNA expressions., Results: Compared with the normal group, rats in the model group had increased pathological damage degree and inflammatory cell infiltration in the colon tissue, slowed-down body weight gain, decreased colon length, spleen weight, serum anti-inflammatory factors IL-2 and IL-10 contents, colonic Arg1/CD68 fluorescence positive expression, and Arg1 protein and mRNA expressions( P <0.01, P <0.05), as well as increased DAI scores, colon histopathological scores, contents of serum pro-inflammatory factors TNF-α and IL-1β, colonic iNOS/CD68 fluorescence positive expression, iNOS protein and mRNA expressions( P <0.01). Compared with the model group, the above indicators were significantly improved in rats of the EA group and the western medication group( P <0.01, P <0.05)., Conclusions: EA of "Changbing Decoction" can improve UC of rats by regulating the polarization of colonic macrophages, inhibiting the generation of M1 macrophages and promoting the generation of M2 macrophages.

Objective: To observe the effect of electroacupuncture （EA） at "Zhongwan" （CV12）, "Tianshu" （ST25） and "Shangjuxu" （ST37） （an acupoint prescription "Changbingfang" for treatment of intestinal disorders） on autophagy and expression of AMPK/mTOR signaling pathway in rats with ulcerative colitis （UC）, so as to explore its mechanism underlying improvement of UC., Methods: Thirty-two male SD rats were randomly divided into control, model, medication and EA groups, with 8 rats in each group. The UC model was established by free drinking of 5% dextran sulfate sodium salt solution for 7 days. EA stimulation （10 Hz/50 Hz） was delivered to CV12, ST25 and ST37 for 20 min, once a day for 3 consecutive days. Rats of the medication group received gavage of mesalazine suspension （200 mg/kg） once a day, 3 times in total. The rats' general conditions were recorded for calculating the disease activity index （DAI） score （0-4 points）. Histomorphological changes of colon were observed via HE staining. The levels of serum interleukin 6 （IL-6）, tumor necrosis factor- α （TNF- α ） and IL-10 were measured by ELISA. The mRNA expressions of LC3B and p62 were tested by fluorescence quantitative PCR. Western blot was used to detect the expression levels of LC3B, p62 and AMPK/mTOR pathway related proteins in colon tissues., Results: Compared with the control group, the DAI score, contents of serum IL-6 and TNF- α , the expression levels of p62 protein and mRNA, ratio of p-mTOR/mTOR were significantly increased （ P <0.01）； while the content of serum IL-10, the expression levels of LC3B mRNA, ratio of LC3BⅡ/LC3BⅠ and p-AMPK/AMPK were decreased （ P <0.01, P <0.05） in the model group. Relevant to the model group, modeling-induced increases of DAI score, serum IL-6, TNF- α and IL-10 contents, expressions of p62 protein and mRNA, LC3B mRNA, ratio of p-mTOR/mTOR, LC3BⅡ/LC3BⅠ and p-AMPK/AMPK were reversed in both medication and EA groups （ P <0.01, P <0.05）. The effect of EA was apparently superior to that of mesalazine in up-regulating ratio of LC3BⅡ/LC3BⅠ and p-AMPK/AMPK, p62 mRNA expression （ P <0.01, P <0.05）, and in down-regulating ratio of p-mTOR/mTOR （ P <0.05）. H.E. staining showed severe damage of the colonic mucosal barrier with infiltration of a large number of inflammatory cells in the model group, which was milder in medication and EA groups., Conclusion: EA of acupoint recipe "Changbingfang" can improve the symptoms in UC rats, which may be related to its functions in promoting colonic autophagy, increasing AMPK phosphorylation level, and decreasing mTOR phosphorylation level.

Background: Human monoclonal antibodies might offer an important new approach to reduce malaria morbidity and mortality. In the first two parts of a three-part clinical trial, the antimalarial monoclonal antibody CIS43LS conferred high protection against parasitaemia at doses of 20 mg/kg or 40 mg/kg administered intravenously followed by controlled human malaria infection. The ability of CIS43LS to confer protection at lower doses or by the subcutaneous route is unknown. We aimed to provide data on the safety and optimisation of dose and route for the human antimalaria monoclonal antibody CIS43LS., Methods: VRC 612 Part C was the third part of a three-part, first-in-human, phase 1, adaptive trial, conducted at the University of Maryland, Baltimore Center for Vaccine Development and Global Health, Baltimore, MD, USA. We enrolled adults aged 18-50 years with no previous malaria vaccinations or infections, in a sequential, dose-escalating manner. Eligible participants received the monoclonal antibody CIS43LS in a single, open-label dose of 1 mg/kg, 5 mg/kg, or 10 mg/kg intravenously, or 5 mg/kg or 10 mg/kg subcutaneously. Participants underwent controlled human malaria infection by the bites of five mosquitoes infected with Plasmodium falciparum 3D7 strain approximately 8 weeks after their monoclonal antibody inoculation. Six additional control participants who did not receive CIS43LS underwent controlled human malaria infection simultaneously. Participants were followed-up daily on days 7-18 and day 21, with qualitative PCR used for P falciparum detection. Participants who tested positive for P falciparum were treated with atovaquone-proguanil and those who remained negative were treated at day 21. Participants were followed-up until 24 weeks after dosing. The primary outcome was safety and tolerability of CIS43LS at each dose level, assessed in the as-treated population. Secondary outcomes included protective efficacy of CIS43LS after controlled human malaria infection. This trial is now complete and is registered with ClinicalTrials.gov, NCT04206332., Findings: Between Sept 1, 2021, and Oct 29, 2021, 47 people were assessed for eligibility and 31 were enrolled (one subsequently withdrew and was replaced) and assigned to receive doses of 1 mg/kg (n=7), 5 mg/kg (n=4), and 10 mg/kg (n=3) intravenously and 5 mg/kg (n=4) and 10 mg/kg (n=4) subcutaneously, or to the control group (n=8). CIS43LS administration was safe and well tolerated; no serious adverse events occurred. CIS43LS protected 18 (82%) of 22 participants who received a dose. No participants developed parasitaemia following dosing at 5 mg/kg intravenously or subcutaneously, or at 10 mg/kg intravenously or subcutaneously. All six control participants and four of seven participants dosed at 1 mg/kg intravenously developed parasitaemia after controlled human malaria infection., Interpretation: CIS43LS was safe and well tolerated, and conferred protection against P falciparum at low doses and by the subcutaneous route, providing evidence that this approach might be useful to prevent malaria across several clinical use cases., Funding: National Institute of Allergy and Infectious Diseases, National Institutes of Health., Competing Interests: Declaration of interests AHI and RAS are listed as inventors on a pending patent application describing CIS43 and related antibodies. All other authors declare no competing interests., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Background: New approaches for the prevention and elimination of malaria, a leading cause of illness and death among infants and young children globally, are needed., Methods: We conducted a phase 1 clinical trial to assess the safety and pharmacokinetics of L9LS, a next-generation antimalarial monoclonal antibody, and its protective efficacy against controlled human malaria infection in healthy adults who had never had malaria or received a vaccine for malaria. The participants received L9LS either intravenously or subcutaneously at a dose of 1 mg, 5 mg, or 20 mg per kilogram of body weight. Within 2 to 6 weeks after the administration of L9LS, both the participants who received L9LS and the control participants underwent controlled human malaria infection in which they were exposed to mosquitoes carrying Plasmodium falciparum (3D7 strain)., Results: No safety concerns were identified. L9LS had an estimated half-life of 56 days, and it had dose linearity, with the highest mean (±SD) maximum serum concentration (C max ) of 914.2±146.5 μg per milliliter observed in participants who had received 20 mg per kilogram intravenously and the lowest mean C max of 41.5±4.7 μg per milliliter observed in those who had received 1 mg per kilogram intravenously; the mean C max was 164.8±31.1 in the participants who had received 5 mg per kilogram intravenously and 68.9±22.3 in those who had received 5 mg per kilogram subcutaneously. A total of 17 L9LS recipients and 6 control participants underwent controlled human malaria infection. Of the 17 participants who received a single dose of L9LS, 15 (88%) were protected after controlled human malaria infection. Parasitemia did not develop in any of the participants who received 5 or 20 mg per kilogram of intravenous L9LS. Parasitemia developed in 1 of 5 participants who received 1 mg per kilogram intravenously, 1 of 5 participants who received 5 mg per kilogram subcutaneously, and all 6 control participants through 21 days after the controlled human malaria infection. Protection conferred by L9LS was seen at serum concentrations as low as 9.2 μg per milliliter., Conclusions: In this small trial, L9LS administered intravenously or subcutaneously protected recipients against malaria after controlled infection, without evident safety concerns. (Funded by the National Institute of Allergy and Infectious Diseases; VRC 614 ClinicalTrials.gov number, NCT05019729.)., (Copyright © 2022 Massachusetts Medical Society.)

Background: Acute-on-chronic liver failure (ACLF) patients have high mortality in a short period of time. This study aimed to compare the prognosis of transplanted ACLF patients to that of nontransplanted ACLF patients and decompensated cirrhosis recipients., Methods: Clinical data of 29 transplanted ACLF patients, 312 nontransplanted ACLF patients, and 60 transplanted decompensated cirrhosis patients were retrospectively collected. Propensity score matching (PSM) analysis was used to match patients between different groups., Results: After PSM, the 90-day and 1-year survival of transplanted ACLF patients was significantly longer than that of nontransplant controls. Although the 90-day survival and 1-year survival of ACLF recipients was similar to that of decompensated cirrhosis controls, ACLF recipients were found to have longer mechanical ventilation, longer intensive care unit (ICU) stay, longer hospital stay, higher incidence of tracheotomy, higher expense, and higher morbidity of complication than matched decompensated cirrhosis controls. The 90-day and 1-year survival of transplanted ACLF grade 2-3 patients was also significantly longer than that of nontransplanted controls., Conclusions: Liver transplantation can strongly improve the prognosis of ACLF patients. Despite having more burdens (including longer mechanical ventilation, longer ICU stay, higher incidence of tracheotomy, longer hospital stay, higher hospitalization expense, and higher complication morbidity), ACLF recipients can obtain similar short-term and long-term survival to decompensated cirrhosis recipients. For severe ACLF patients, liver transplantation can also significantly improve their short-term and long-term survival., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Chen, Wu, Huang, Wang, Zheng, Yu, Wang, Hou, Ye, Zhang and Zhao.)

Background: The studies of laparoscopic-assisted transhiatal gastrectomy (LTG) in patients with Siewert type II adenocarcinoma of the esophagogastric junction (AEG) are scarce., Aim: To compare the surgical efficiency of LTG with the open transhiatal gastrectomy (OTG) for patients with Siewert type II AEG., Methods: We retrospectively evaluated a total of 578 patients with Siewert type II AEG who have undergone LTG or OTG at the First Medical Center of the Chinese People's Liberation Army General Hospital from January 2014 to December 2019. The short-term and long-term outcomes were compared between the LTG ( n = 382) and OTG ( n = 196) groups., Results: Compared with the OTG group, the LTG group had a longer operative time but less blood loss, shorter length of abdominal incision and an increased number of harvested lymph nodes ( P < 0.05). Patients in the LTG group were able to eat liquid food, ambulate, expel flatus and discharge sooner than the OTG group ( P < 0.05). No significant differences were found in postoperative complications and R0 resection. The 3-year overall survival and disease-free survival performed better in the LTG group compared with that in the OTG group (88.2% vs 79.2%, P = 0.011; 79.7% vs 73.0%, P = 0.002, respectively). In the stratified analysis, both overall survival and disease-free survival were better in the LTG group than those in the OTG group for stage II/III patients ( P < 0.05) but not for stage I patients., Conclusion: For patients with Siewert type II AEG, LTG is associated with better short-term outcomes and similar oncology safety. In addition, patients with advanced stage AEG may benefit more from LTG in the long-term outcomes., Competing Interests: Conflict-of-interest statement: All authors declare that there is no conflict of interest., (©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.)

Background: Diagnosis of patients with hymenoptera venom hypersensitivity consists of elucidating clinical symptoms suggestive of systemic reaction (SR) and then confirmation of sensitization via intradermal skin testing (IDST) first and serum IgE assays such as ImmunoCAP (ICAP) as a complementary modality of diagnosis., Objective: Determine the concordance between ICAP and IDST in patients with a clinical history suggestive of hymenoptera venom SR. Determine whether venom immunotherapy would change on the basis of IDST versus ICAP results., Methods: A prospective diagnostic study was designed to test the concordance between IDST and ICAP venom testing in the diagnosis of hymenoptera venom hypersensitivity. This study entailed testing both IDST and ICAP for 5 hymenoptera venoms (honey bee, wasp, yellow jacket, yellow hornet, and white-faced hornet) in both a case group with SR to hymenoptera venom (N = 70) and a control group without SR (N = 51)., Results: Significant discordance was observed between positive IDST and ICAP results for any of the 5 hymenoptera venoms (McNemar test, P = .001). In the case group, there was significant discordance for wasp (P < .0001), yellow jacket (P = .002), and white-faced hornet (P = .02). More than 47% of the case patients would have different venom immunotherapy prescriptions if ICAP and IDST had been performed during initial diagnosis versus IDST alone., Conclusions: Our study shows significant discordance between IDST and ICAP; however, they are complementary. On the basis of our data, we propose ICAP testing first followed by IDST for ICAP-negative venoms as an alternative and efficient diagnostic strategy., (Published by Elsevier Inc.)

Acute-on-chronic liver failure(ACLF) is the most severe form of acute decompensation that develops in patients with chronic liver disease or liver cirrhosis,and is always accompanied by one or more extrahepatic organ failure, and has an extremely poor short-term prognosis. The causes triggering ACLF are complex and diverse,and the clinical stage and the type and the definition of organ failure differ greatly from one another. Therefore, a universally accepted diagnostic criteria for ACLF is not to be defined, and the epidemiological data and patient outcomes on ACLF are not easy to predict and compare among different regions. Accumulating evidence has shown that liver transplantation(LT) plays a significant role in the surgical treatment of patients with ACLF,but its clinical value is still controversial. The specific management and treatment strategy after the admission of patients with ACLF has not yet formed a unified and standardized process or opinions, which includes the monitoring in the ICU,the support and maintenance of organ functions, the selection of the surgical indication and the timing for LT and so on. Moreover, there still exists many controversies concerning, for example, whether patients with ACLF should receive greater priority for organ allocation compared to other potential candidates on the waiting list. Besides, more prospective controlled studies are urgently needed to investigate the role of the artificial liver support system in the bridging therapy to LT. The aim of this article is to review the indication selection of patients with ACLF suitable for LT,the survival outcomes and prognostic factors after LT, the selection of timing, the organ allocation policy and the bridging therapy to LT, which intends to provide new direction for designing the future clinical studies on LT in patients with ACLF.

Background: Additional interventions are needed to reduce the morbidity and mortality caused by malaria., Methods: We conducted a two-part, phase 1 clinical trial to assess the safety and pharmacokinetics of CIS43LS, an antimalarial monoclonal antibody with an extended half-life, and its efficacy against infection with Plasmodium falciparum . Part A of the trial assessed the safety, initial side-effect profile, and pharmacokinetics of CIS43LS in healthy adults who had never had malaria. Participants received CIS43LS subcutaneously or intravenously at one of three escalating dose levels. A subgroup of participants from Part A continued to Part B, and some received a second CIS43LS infusion. Additional participants were enrolled in Part B and received CIS43LS intravenously. To assess the protective efficacy of CIS43LS, some participants underwent controlled human malaria infection in which they were exposed to mosquitoes carrying P. falciparum sporozoites 4 to 36 weeks after administration of CIS43LS., Results: A total of 25 participants received CIS43LS at a dose of 5 mg per kilogram of body weight, 20 mg per kilogram, or 40 mg per kilogram, and 4 of the 25 participants received a second dose (20 mg per kilogram regardless of initial dose). No safety concerns were identified. We observed dose-dependent increases in CIS43LS serum concentrations, with a half-life of 56 days. None of the 9 participants who received CIS43LS, as compared with 5 of 6 control participants who did not receive CIS43LS, had parasitemia according to polymerase-chain-reaction testing through 21 days after controlled human malaria infection. Two participants who received 40 mg per kilogram of CIS43LS and underwent controlled human malaria infection approximately 36 weeks later had no parasitemia, with serum concentrations of CIS43LS of 46 and 57 μg per milliliter at the time of controlled human malaria infection., Conclusions: Among adults who had never had malaria infection or vaccination, administration of the long-acting monoclonal antibody CIS43LS prevented malaria after controlled infection. (Funded by the National Institute of Allergy and Infectious Diseases; VRC 612 ClinicalTrials.gov number, NCT04206332.)., (Copyright © 2021 Massachusetts Medical Society.)

Immune checkpoint inhibitors are increasingly used in the treatment of various solid tumors, including hepatocellular carcinoma (HCC). For liver transplant recipients, the safety of using immune checkpoint inhibitors before or after transplantation remains to be further explored. Former reports were mainly about posttransplant use of immune checkpoint inhibitors resulting in allograft rejection. Here we present one HCC patient who received toripalimab (an immune checkpoint inhibitor currently in phase 3 clinical trial for HCC) therapy before undergoing liver transplantation. He finally suffered fatal acute hepatic necrosis which is likely to be related to the acute immune rejection caused by the pretransplant use of toripalimab., (Copyright © 2021. Published by Elsevier B.V.)

Objectives: The aim of this study was to investigate the association between survival of anti-MDA5 autoantibody-positive/negative patients with myositis-associated interstitial lung disease (MA-ILD) and neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), monocyte-lymphocyte ratio (MLR), C-reactive protein-albumin ratio (CAR), and erythrocyte sedimentation rate-albumin ratio (EAR). Method: The study included 104 patients diagnosed with MA-ILD between January 2017 and February 2019 at the First Affiliated Hospital, University of Science and Technology of China. The clinical and laboratory results were compared between survivors and non-survivors in anti-MDA5 autoantibody-positive and anti-MDA5 autoantibody-negative patients. Cox proportional hazard models were used for univariable and multivariate analyses to determine survival-related factors. A logistic regression model was used to establish a joint diagnosis, and the feasibility of the combined diagnosis to evaluate the prognosis of MA-ILD was explored. Results: Among 47 anti-MDA5-positive patients with MA-ILD, EAR was an independent predictor of survival. When separated into high and low subgroups, high MLR (> 0.604) and EAR (> 1.458) were predictive of survival (p < 0.05). High MLR, high EAR, and age combined with lactate dehydrogenase were the highest (0.886) in predicting the prognosis of MA-ILD, and were higher than the area under the curve diagnosed separately. In 57 anti-MDA5-negative patients with MA-ILD, NLR and high EAR (> 0.872) were independent predictors of survival (p < 0.05). Conclusion: MLR and EAR are associated with prognosis in anti-MDA5-positive patients. NLR and EAR are associated with prognosis in anti-MDA5-negative patients. Using NLR, MLR, and EAR, inflammatory conditions of MA-ILD can be predicted and possible outcomes estimated.

In this work, a rhamnolipid (RL) pretreatment technology was proposed to promote methane production from two-phase anaerobic digestion of waste activated sludge. In the first phase (i.e., acidogenic phase), the WAS hydrolysis and acidogenesis were significantly enhanced after RL pretreatment for 4 day, under which the concentration of soluble protein and the short-chain fatty acids (SCFA) in the presence of RL at 0.04 g/g TSS was respectively 2.50 and 5.02 times higher than that without RL pretreatment. However, methane production was inhibited in the presence of RL. In the second phase (i.e., methanogenic phase), batch biochemical methane potential tests suggested that the addition of RL is effective in promoting anaerobic methane production. With an increase of RL dosage from 0 to 0.04 g/g TSS, the cumulative methane yield increased from 100.42 ± 3.01 to 168.90 ± 5.42 mL. Although the added RL could be utilized to produce methane, it was not the major contributor to the enhancement of methane yield. Further analysis revealed that total cumulative yield from the entire two-phase anaerobic digestion (sum of the yield of the acidogenic phase and methanogenic phase) increased from 113.42 ± 3.56 to 164.18 ± 5.20 mL when RL dosage increased from 0 to 0.03 g/g TSS, indicating that the addition of RL induced positive effect on the methane production of the entire two-phase anaerobic digestion. The enzyme activity analysis showed that although higher dosages of RL still inhibited the microorganisms related to methanogenesis to some extends in the methanogenic phase, the inhibitory effect was significantly weakened compared to the acidogenic phase. Microbial analysis revealed that RL reduced the abundance of Candidatus&#95;Methanofastidiosum sp. while increased the abundance of Methanosaeta sp., causing the major methanogenesis pathway to change from hydrogenotrophic to aceticlastic. Moreover, the community of hydrolytic microbes and acidogens was shifted in the direction that is conducive to hydrolysis-acidogenesis. The findings reported not only expand the application field of RL, but also may provide supports for sustainable operation of wastewater treatment plants (WWTPs)., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier Ltd.)

Neonicotinoid insecticides (NNIs) are the most widely used insecticides in China and worldwide. Continuous use of NNIs can lead to their accumulation in soil, causing potential ecological risks due to their relatively long half-life. We used liquid chromatography-tandem mass spectrometry (LC-MS/MS) to investigate the residual levels of nine neonicotinoids in greenhouse soils in Shouguang, East China, at different soil depths and with different crops (tomato and cucumber) after varying periods of cultivation. Seven neonicotinoids were detected in the soils of the tomato greenhouses and six were detected in the soils of the cucumber greenhouses, with total concentrations ranging from 0.731 to 11.383 μg kg -1 and 0.363 to 19.224 μg kg -1 , respectively. In all samples, the neonicotinoid residues in the soils cultivated for 8-9 years were lower than in those cultivated for 2 years and 14-17 years. In the tomato greenhouse soils, the residual levels of NNIs were highest in the topsoil, with progressively lower concentrations found with depth. Under cucumber cultivation, the NNI residue levels were also highest in the topsoil but there was little difference between the middle and lower soil layers. Total organic carbon (TOC) decreased with soil depth while pH showed the opposite trend, showing a significant negative correlation in both types of soils (tomato soils ρ = -0.900, p = .001; cucumber soils ρ = -0.883, p = .002). Furthermore, TOC was significantly positively correlated, and pH was negatively correlated, with total NNI concentrations in both types of soils (TOC: tomato soils ρ = 0.800, p = .010; cucumber soils ρ = 0.881, p = .004; pH: tomato soils ρ = -0.850, p = .004; cucumber soils ρ = -0.643, p = .086). The results of an ecological risk analysis showed that acetamiprid represents a particularly high toxicity risk in these soils. Based on our analysis, NNI residues in the soils of tomato greenhouses and their associated ecological risks deserve more attention than those of cucumber greenhouse soils., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The understanding of soil microbiome is important for sustainable cultivation, especially under greenhouse conditions. Here, we investigated the changes in soil pesticide residues and microbial diversity and community structure at different cultivation years under a greenhouse system. The 9-to-14 years sites were found to have the least diversity/rich microbial population as compared to sites under 8 years and over 16 years, as analyzed with alpha diversity index. In total, 42 bacterial phyla were identified across soils with different pesticide residues and cultivation ages. Proteobacteria, Acidobacteria, and Bacteroidetes represented the dominant phyla, that accounted for 34.2-43.4%, 9.7-19.3% and 9.2-16.5% of the total population, respectively. Our data prove that certain pesticides contribute to variation in soil microbial community and that soil bacteria respond differently to cultivation years under greenhouse conditions. Thus, this study provides an insight into microbial community structure changes by pesticides under greenhouse systems and natural biodegradation may have an important part in pesticides soil decontamination., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Objective: To evaluate the early predictive value of immature granulocytes(IGs) in postoperative infection in patients with spinal injury., Methods: The clinical data of 78 patients with spinal cord injury underwent surgery from October 2016 to October 2018 were collected. There were 57 males and 21 females, aged from 45 to 63 years old with an average of (55.2±6.1) years. The most common cause of injury was spinal trauma or degeneration. According to the American Spinal Cord Injury Association (ASIA) classification of spinal cord injury, 37 cases were complete injury(grade A and B) and 41 cases were incomplete injury (grade C and D). All the 78 patients showed different degrees of deep and shallow sensory disturbances, and underwent CT and MRI examinations after admission. According to whether the secondary infection occurred within 1 week after surgery, the patients were divided into infected group and non-infected group. At the time of admission and 1, 3, 6 days after surgery, the immature granulocyte absolute value(IG#), immature granulocyte percentage(IG%), C-reactive protein(CRP), procalcitonin(PCT), interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-α (TNF-α) were detected. Using ROC curve to compare the predictive value of IG%, IG#, and PCT in early detection of postoperative infection. The correlation between IG%, IG# and PCT was analyzed using the Spearman correlation test., Results: At a week after operation, 33 patients occurred infection and 45 patients occurred no infection(infected group and non-infected group). All patients were followed up to no serious complications and deaths at the time of discharge. The proportion of complete injury in the infected group was significantly higher than that in the non-infected group( t= 3.979, P= 0.046), and the cervical and thoracic vertebrae were more common in the infected group, and the non-infected group was mostly lumbar injury( t= 6.226, P= 0.044). One day after surgery, PCT, IG%, IG# were resectively(0.71±0.10) ng/ml, 1.08±0.10, 0.20±0.05 in infected group, while in non-infected group were(0.51±0.08) ng/ml, 0.82±0.13, 0.13±0.04, there was significant difference between two groups( P <0.001). At 3 days postoperatively, CRP, PCT, IL-6, IL-1β, TNF-α, IG%, IG# were resectively (80.47±15.74) mg/L, (2.39±0.27) ng/ml, (15.74±3.85) pg/ml, (16.47±4.75) pg/ml, (2.18±0.57) ng/ml, 0.28±0.10, 1.38±0.54 in injected group, while in non-infected group were(62.42±14.68) mg/L, (0.89±0.21) ng/ml, (13.10±3.87)pg/ml, (14.57±3.35) pg/ml, (1.63±0.37) ng/ml, 0.09±0.01, 0.83±0.24, there was significant difference between two groups( P <0.001). At 6 days postoperatively, the laboratory parameters of the infected group were significantly higher than those of the non-infected group( P <0.001). ROC curve analysis showed that PCT, IG%, and IG# could predict early infection after spinal cord injury( P <0.001), and the area under the curve (AUC) of IG# was significantly lower than PCT and IG%(respectively 0.847, 0.947, 0.934, P <0.05). Correlation analysis showed that IG%, IG# and PCT were significantly correlated, and the correlation coefficients were 0.724 and 0.472, respectively( P <0.001)., Conclusions: The elevated levels of IG%, IG# and PCT in patients with spinal cord injury within 24 hours after surgery have high sensitivity and specificity for predicting early secondary infection. Postoperative quantification of these indicators helps early identification of patients with high risk of infection., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© 2019 by the China Journal of Orthopaedics and Traumatology Press.)

The binding constant (K doc ) of organic pollutants (OPs) with dissolved organic matter (DOM) is an important parameter in determining the partitioning of OPs in the aquatic environment. Most estimation models have focused on calculating the K doc of a specific group of OPs but failed to obtain K doc values of different OPs effectively over the last three decades. In this study, we attempted to build a general-applicable K doc model based on various organic compounds' K doc values from the literature since 1973. Two multiple linear regression models, a DOM nonspecific model and an Aldrich HA model, were developed based on two solid and easy to access parameters-molecular connectivity indices (MCI) and polarity correction factors (PCF). In addition, the models' corresponding K ow -K doc models, which were mostly used in previous model studies, were developed for comparison. The adjusted determining coefficient (adj-R 2 ) and standard error of the estimate (SEE) of the DOM nonspecific MCI-PCF-K doc model were 0.815 and 0.579, respectively, whereas the adj-R 2 and SEE for the MCI-PCF-K doc model of Aldrich HA reached 0.907 and 0.438, respectively. The Aldrich HA model showed higher pertinence to the nonspecific model. Furthermore, both models exhibited better fit than the K ow -K doc models. The dipole moment modification attempts did not significantly improve either MCI-PCF-K doc models; hence, the two models were not altered with the dipole moment. The robustness tests by a Jackknifed method showed that the two MCI-PCF-K doc models exhibited higher robustness than the K ow -K doc . Of all of the OPs, the phenols contributed the most to their robustness. Furthermore, a sensitivity analysis showed that the two MCI-PCF-K doc models were sensitive to the robust parameters., (Copyright © 2019 Elsevier B.V. All rights reserved.)

On the basis of tight-binding approximation, the energy absorption of 2D atomic clusters is calculated by the linear response theory. Through the energy-absorption peaks in the presence of different external potentials, various types of plasmon modes are specified in clusters with dozens to hundreds atoms, such as transverse dipole plasmon, longitudinal dipole plasmon, transverse quadrupole plasmon, and longitudinal quadrupole plasmon. Moreover, the transformation of plasmon from quantum to classical mode is observed in clusters with different shape and different electron density. The particular transformation process demonstrate that: there are only a few modes of plasmon in clusters with few-atoms; as the number of atoms in cluster is increased, the number of plasmon modes increases, the gaps between plasmon frequencies become smaller, the quantum modes of plasmon gradually evolve into continuous modes, and the dispersion of quantum-mode plasmon gradually transforms into the one of classical 2D plasmon. Such process reveals the size effect of plasmon in 2D clusters, which can be explained by the fact that the energy levels near the Fermi energy are denser and more compact in larger-size clusters.

Objective: To explore the function and role of innate lymphoid cells in the pathogenesis of systemic lupus erythematosus (SLE) at different disease activity levels. Methods: From Nov 2017 to May 2018, 40 patients with SLE and 15 age-matched healthy non-immune-related diseases controls were enrolled from Anhui provincial hospital. According to the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI)-2K, the patients were divided into active group ( n= 20) and remission group ( n= 20). The frequency of ILCs, B cells, CD4+T and CD8+T cells from peripheral blood mononuclear cells (PBMCs) was detected by flow cytometry. The subsets of ILCs in each group were compared with the subsets of B cells and T cell respectively. The levels of IL-4, IL-33 and IFN-γ in each group were tested by ELISA. Result: Compared with the control group, ILC1 percentage was significantly increased in SLE active group [(22.33%±2.52%) vs (14.56%±1.28%), P= 0.018 1]; ILC2 percentage was decreased significantly in both remission group [(19.67%±1.83%) vs (42.48%±3.46%), P< 0.000 1] and active group [(8.67%±0.83%) vs (19.67%±1.83%), P< 0.000 1]; ILC3 percentage was decreased significantly in active group [(5.72%±1.08%) vs (14.35%±2.40%), P= 0.001 3]. SLEDAI score was negatively correlated with the percentage of ILC2 ( P= 0.023 9) in all patients. The percentage of ILCs in the remission group ( P= 0.046 2) and activity group ( P= 0.003 7) were both increased significantly. Moreover, the percentage of ILC2 in active group was negatively correlated with CD4+T cells ( P= 0.030 8), and the serum IgG was negatively correlated with ILC2% in all patients ( P= 0.013 8). Compared with control group or remission group, the levels of IFN-γ ( F= 10.91, P= 0.000 1) and IL-4 ( F= 6.046, P= 0.004 7) in active group were remarkable higher. However, IL-33 was significantly reduced in active group ( F= 6.645, P= 0.002 7). The percentage of ILC2 ( r= 0.154 3, P= 0.028 8) and ILC3 ( r= 0.313 6, P= 0.001 1) in all patients with SLE were positively correlated with the level of IL-4. Conclusion: The percentage of ILCs is related to disease activity, and ILCs play a "double-edged" role in the pathogenesis of SLE. Its function and mechanism are worth further exploration.

BMP4/7-dependent expression of inhibitor of differentiation/DNA binding (Id) proteins 1 and 3 has been implicated in tumor progression and poor prognosis of malignant melanoma patients. Hyaluronic acid (HA), a pericellular matrix component, supports BMP7 signalling in murine chondrocytes through its receptor CD44. However, its role in regulating BMP signalling in melanoma is not clear. In this study we found that depletion of endogenously-produced HA by hyaluronidase treatment or by inhibition of HA synthesis by 4-methylumbelliferone (4-MU) resulted in reduced BMP4/7-dependent Id1/3 protein expression in mouse melanoma B16-F10 and Ret cells. Conversely, exogenous HA treatment increased BMP4/7-dependent Id1/3 protein expression. Knockdown of CD44 reduced BMP4/7-dependent Id1/3 protein expression, and attenuated the ability of exogenous HA to stimulate Id1 and Id3 expression in response to BMP. Co-IP experiments demonstrated that CD44 can physically associate with the BMP type II receptor (BMPR) ACVR2B. Importantly, we found that coordinate expression of Id1 or Id3 with HA synthases HAS2, HAS3, and CD44 is associated with reduced overall survival of cutaneous melanoma patients. Our results suggest that HA-CD44 interactions with BMPR promote BMP4/7-dependent Id1/3 protein expression in melanoma, contributing to reduced survival in melanoma patients.

Objective: To analyze the effect of warming needle moxibustion of Jiaji (EX-B 2), Huantiao (GB 30), etc. on lumbago and other symptoms of lumbar disc herniation (LDH) patients and changes of plasma β-endorphin (β-EP) content., Methods: A total of 60 LDH patients were equally randomized into warming needle moxibustion group and conventional acupuncture group. Patients of the conventional acupuncture group were treated by puncturing lumbar Jiaji (EX-B 2), Huantiao (GB 30), Weizhong (BL 40), and Yanglingquan (GB 34) with filiform acupuncture needles. Patients of the warming needle moxibustion group were treated by puncturing the same 4 acupoints, and with the acupuncture needle in lumbar EX-B 2 attached an ignited moxa-stick segment. The treatment in both groups lasted for 30 min every time, once every other day for 15 times, with 5 times being a therapeutic course and two days' rest between every two courses. The therapeutic effect was evaluated according to the "Criteria for Evaluating the Therapeutic Effect of Disorders of Traditional Chinese Medicine". The lumbago severity was assessed using visual analogue scale (VAS), and the lumbar functional activity assessed using modified Oswestry dysfunction (in objects-lifting, sitting, standing, walking, sleeping, pain, activity of daily living, social activity, outing, etc.) index (ODI). The plasma β-EP content was assayed using ELISA., Results: After the treatment, of the two 30 cases in the conventional acupuncture and warming needle moxibustion groups, 8 (26.67%) and 12 (40.00%) were under control, 8 (26.67%) and 10 (33.33%) had a marked improvement, 3 (10.00%) and 4 (13.33%) were effective, and 11 (36.66%) and 4 (13.33%) ineffective, with the effective rates being 63.34% and 86.66%, respectively. The effective rate of the warming needle moxibustion was significantly higher than that of the conventional acupuncture ( P <0.05). The VAS scores and ODI values of the two groups were gradually decreased along with the prolongation of treatment, and significantly lower in the warming needle moxibustion group than in the acupuncture group at the 1 st , 2 nd and 3 rd course of treatment ( P <0.01). The levels of plasma β-EP were gradually increased in the two groups at the 1 st , 2 nd and 3 rd course of treatment, and were obviously higher in the warming needle moxibustion group than in the conventional acupuncture group at each of the 3 courses ( P <0.01). No obvious adverse reactions were observed in the two groups., Conclusion: Warming needle moxibustion has a better therapeutic effect in relieving lumbago and lumbar dysfunction, and can up-regulate blood β-EP level.

Based on the self-consistent linear response theory, the plasmon-energy absorption in linear atomic chain are studied by using the tight-binding approximation. Results indicate that the eigen-frequency of the plasmon is uninfluenced by the external electric potential, but the plasmon modes excited by various electric potentials are obviously different. Each mode of plasmon corresponds to one kind of eigen-charge distribution. When the plasmon mode is excited, the resonant charge will show a distribution characteristic the same as the one of eigen charge. And the plasmon mode can be precisely controlled by external electric potential if the eigen-charge distribution at such plasmon is known. The relationship between plasmon-energy absorption and atom number are also affected by the external electric potential. However, most of the other studies only show the normal case that the plasmon-energy absorption increases with the atom number increasing. Here, we demonstrate that the normal case commonly occurs under monotone increasing potential. And abnormal case may occur under monotone decreasing potential, ie, the plasmon-energy absorption will decrease with the atom number increasing. But, in the presence of arbitrary potential applied to the same atomic chain, the plasmon-energy absorption will always increase with the electron number increasing.

Objective: To investigate the roles of BDNF/TrkB neurotrophic signaling in hippocampal injury for fatigue rats induced by incremental load exercise and the protective effects and mechanism of spirulina supplement., Methods: Sixty SD rats were randomly divided into normal control group (NC), normal plus spirulina group(NS), exercise model group (EM), exercise plus spirulina group (ES), and positive control group (PC), 12 rats in each group.Group EM, Group ES and Group PC were applied by treadmill running with high-intensity increasing for three weeks, and Group NC had not any intervention measures.Group ES and Group NS were treated with spirulina at a dose of 300 mg/kg.bw.by intragastric administration.Group PC was gavaged at the same volume of ginseng extract of 1.92 g/kg for three weeks.The expressions of brain-derived neurotrophic factor (BDNF), tyrosine kinase recptor (TrkB), phospho-tyrosine kinase recptor (p-TrkB) were tested by Western blot and immunohistochemical method, and micromorphology changes of hippocampal CA1 were observed by light microscope at the end of the experiment.The general situations of rats such as body weights were recorded during the experiment., Results: Compared with Group NC, Group EM showed significantly decrease in body weight and hippocampal CA1 neurons of the group loosely arrayed and disarrayed and some neurons were shrinked, and even some neurons disappeared.The expressions of BDNF, TrkB and p-TrkB in group EM were increased significantly( P <0.01).Compared with Group EM, body weight of Group ES was increased significantly, and the above mentioned injuries of neurons were improved significantly:the number of neurons and nissl bodies were significantly increased and the neurons arrayed regularly and its morphology was more complete.The expressions of BDNF, TrkB and p-TrkB in the group were increased significantly( P <0.05 or P <0.01).And there was no difference between Group ES and Group PC., Conclusions: BDNF/TrkB neurotrophic signal pathway could be involved in the repair process of hippocampal nervous damage caused by incremental load exercise for fatigue rats.Spirulina supplement had a protective effect on the damaged nervous through increasing the expressions of BDNF, TrkB and p-TrkB.

Cytochrome P450 14α-sterol demethylase (CYP51), the key enzyme in sterol biosynthesis pathway, is an important target protein of cholesterol-lowering agents, antifungal drugs, and herbicides. CYP51B enzyme is one of the CYP51 family members. In the present study, we have chosen four representative inhibitors of CYP51B: diniconazole (Din), fluconazole (Flu), tebuconazole (Teb), and voriconazole (Vor), and launched to investigate the binding features of CYP51B-inhibitor and gating characteristics via molecular docking and molecular dynamics (MD) simulations. The results show that the ranking of binding affinities among these inhibitors is Din > Teb > Vor > Flu. Din shows the strongest binding affinity, whereas Flu shows the weakest binding affinity. More importantly, based on the calculated binding free energy results, we hypothesize that the nonpolar interactions are the most important contributors, and three key residues (Thr77, Ala258, and Lys454) play crucial role in protein-inhibitor binding. Besides, residue Phe180 may play a molecular switch role in the process of the CYP51B-Teb and CYP51B-Vor binding. Additionally, Tunnel analysis results show that the major tunnel of Din, Flu, and Teb is located between helix K, FG loop, and β4 hairpin (Tunnel II).The top ranked possible tunnel (Tunnel II) corresponds to Vor exits through helix K, F and helix J. This study further revealed the CYP51B relevant structural characteristics at the atomic level as well as provided a basis for rational design of new and more efficacious antifungal agents.

The present study aims to explore the effectiveness of decompressive craniectomy with bifrontal coronal incision in the management of severe contusion and laceration of bilateral fronto-temporal lobes, as well as the outcomes of early cranioplasty. The authors performed the bifrontal decompressive craniectomy on 56 patients with contusion and laceration of bilateral frontal and temporal lobes, and their follow-up treatment outcomes were tracked within 6 months using Glasgow Outcome Scale. The results showed that 33 patients (out of 56, 58.9%) have recovered, 12 patients (out of 56, 21.4%) have moderate defects, 5 patients (out of 56, 8.9%) have severe defects, 3 patients (out of 56, 5.3%) stayed in persistent vegetative status, and the remaining 3 patients (out of 56, 5.3%) have been dead. There was no persistent temporal hollowing. No patients required revision surgery with modified titanium mesh in this study. Particularly, 28 patients have successfully accepted the early cranioplasty with bone flap or computer-assisted design titanium mesh, and showed good recovery. These results together indicated that the decompressive craniectomy with bifrontal coronal incision in the management of severe contusion and laceration of bilateral fronto-temporal lobes can significantly relieve the comorbidity of intracranial hypertension, and improve the prognosis obviously, thus finally increasing the probability of successful rescue and decreasing the probability of mortality and disability.

Increasing sophistication in methods used to account for human polymorphisms in susceptibility to drug metabolism has been one of the success stories in the prevention of adverse drug reactions. Genetic polymorphisms in drug-metabolizing enzymes can affect enzyme activity and cause differences in treatment response or drug toxicity. CYP2C19 is one of the most highly polymorphic CYP enzymes and acts on 10-15% of drugs in current clinical use. Despite the number of experimental analyses carried out for this system, the detailed structural basis for altered catalytic properties of polymorphic CYP2C19 variants remains unexplained at the atomic level. To this end, we have investigated the mutation effects on structural characteristics and tunnel geometry upon single point mutations to elucidate the underlying molecular mechanism for the enzymatic activity deficiencies by using the fully atomistic molecular dynamics (MD) simulations in their native, membrane-bound cellular environment. The obtained results demonstrate how significant sequence divergence causes heterogeneous variations, and further affects the shape and chemical properties of the substrate binding site. Principal component analysis (PCA) results combined with free energy calculations have revealed distinct mechanisms for different peripheral variants, implying a more complicated process for the decrease/loss of enzymatic activity upon the introduction of point mutations in CYP2C19 rather than simply structural changes of the region where the mutation is located. Overall, our present study provides important insights into the current pharmacogenetic knowledge of human drug-metabolizing CYP2C19 to understand the large inter-individual variability in drug clearance. The knowledge of heterogeneous variations in structural features could guide future experimental and computational work on efficient and safe drug treatment with better pharmacokinetic properties based on the common variant alleles of CYP genes, which varies among different ethnic populations.

The mechanisms underlying diarrhea-predominant irritable bowel syndrome (IBS-D) are poorly understood, but increased intestinal permeability is thought to contribute to symptoms. A recent clinical trial of gluten-free diet (GFD) demonstrated symptomatic improvement, relative to gluten-containing diet (GCD), which was associated with reduced intestinal permeability in non-celiac disease IBS-D patients. The aim of this study was to characterize intestinal epithelial tight junction composition in IBS-D before and after dietary gluten challenge. Biopsies from 27 IBS-D patients (13 GFD and 14 GCD) were examined by H&E staining and semiquantitative immunohistochemistry for phosphorylated myosin II regulatory light chain (MLC), MLC kinase, claudin-2, claudin-8 and claudin-15. Diet-induced changes were assessed and correlated with urinary mannitol excretion (after oral administration). In the small intestine, epithelial MLC phosphorylation was increased or decreased by GCD or GFD, respectively, and this correlated with increased intestinal permeability (P<0.03). Colonocyte expression of the paracellular Na + channel claudin-15 was also markedly augmented following GCD challenge (P<0.05). Conversely, colonic claudin-2 expression correlated with reduced intestinal permeability (P<0.03). Claudin-8 expression was not affected by dietary challenge. These data show that alterations in MLC phosphorylation and claudin-15 and claudin-2 expression are associated with gluten-induced symptomatology and intestinal permeability changes in IBS-D. The results provide new insight into IBS-D mechanisms and can explain permeability responses to gluten challenge in these patients., Competing Interests: The authors declare no conflict of interest.

Purpose: Hyaluronan (HA), an extracellular and peri-cellular glycosaminoglycan with a large molecular weight, plays an important role in cancer growth and metastasis. The aim of this study was to summarize the biological roles and regulation of HA and small HA fragments, and their metabolismn enzymes and receptors in human digestive cancers., Methods: A systematic literature search mainly focusing on the biological roles of HA in the development and progression of human digestive cancers was performed using electronic databases., Results: The correlation between HA accumulation and tumor progression has been shown in various digestive cancers. HA and HA fragment-tumor cell interaction could activate the downstream signaling pathways, promoting cell proliferation, adhesion, migration and invasion, and inducing angiogenesis, lymphangiogenesis, epithelial-mesenchymal transition, stem cell-like property, and chemoradioresistance in digestive cancers., Conclusions: A better insight into the mechanism of HA and HA fragment involvement in digestive cancer progression might be useful for the development of novel biomarkers and therapeutic strategies.

This study aims to develop a method for isolating and purifying protoplasts/vacuoles from fresh leaves of the Cd hyperaccumulator plant species, Sedum alfredii. The results revealed that preheating cellulase and macerozyme at 50 °C for 5 min significantly accelerated the cell wall degradation. For the most optimal conditions for mesophyll protoplast isolation, the mixture of fresh leaves and cell lysates was followed by a 2-h-long vibration. The protoplast lysate for vacuole isolation was diluted, and 0.675 mmol/L was identified as the most appropriate 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonic acid (CHAPS) level, in which S. alfredii large vacuoles are characterized by a high metal and malic acid content. For the best vacuole purification results, we established that 0.8 mol/L was the most optimal mannitol level in the vacuole buffer in terms of vacuole protection during centrifugation, whereas a Ficoll concentration of 0.10 g/ml was adopted in the density-gradient centrifugation., Competing Interests: Compliance with ethics guidelines: Xiao-yu GAO, Xing-cheng LIAO, Ruo-lai WU, Ting LIU, Hai-xing WANG, and Ling-li LU declare that they have no conflict of interest. This article does not contain any studies with human or animal subjects performed by any of the authors.

Aim: The treatment of large (>5 cm) hepatocellular carcinoma (HCC) remains controversial. The aim of this study was to report short and long term outcomes and analyze the factors associated with long term survival for patients who underwent hepatic resection for large HCC., Methods: All patients who underwent hepatic resection for large HCC at the department of Hepato-Pancreato-Biliary Surgery of the First Affiliated Hospital of Anhui Medical University between August 2005 and December 2011 were identified and included for analysis. Demographic and operative data, pathological findings and post-operative outcomes were entered into a computer database. Prognostic factors were analyzed by univariate and multivariate analysis., Results: Ninety-nine patients were included for analysis. Two patients died within 30 days of surgery secondary to hepatic failure. The 1-, 3-, 5-year disease-free survival and overall survival rates following hepatic resection were 67%, 49%, 37% and 77%, 56%, 43%, respectively. Poor histological grade was the only independent predictor of a reduced 5-year disease-free survival. Spontaneous tumor rupture and tumor recurrence were independent predictors of a reduced 5-year overall survival., Conclusions: For selected patients with large HCC, hepatic resection can be performed safely and effectively with moderate expectation of long term survival. True cure however remains rare., (Copyright © 2016 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Two new isoquinoline alkaloids 1-2 and seven known compounds 3-9 were isolated from the ethanol extract of centipede Scolopendra subspinipes mutilans L. Koch. The structures were elucidated by a combination of spectroscopic analyses including 1D and 2D NMR, and HRESIMS. Compounds 1-2 exhibited good cytotoxicity with IC50 values ranging from 1.19 to 31.28μM against six human cancer cell lines and low cytotoxicity against human normal liver L-02 cell lines, suggesting that compounds 1-2 had high specific cytotoxicity on human cancer cell lines. Further analyses showed that compounds 1-2 inhibited U87 cells proliferation by arresting cell cycle progress at G0/G1 phase and inducing apoptosis through loss of mitochondrial membrane potential (MMP), activation of caspase 9/3 and down-regulation of the Bcl-2/Bax protein ratio. The results suggest that compounds 1-2 induce apoptosis in U87 cells through the mitochondria apoptosis pathway, and they deserve further research as potential anti-glioma cancer agents., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Gastric cancer (GC) is one of the most common malignancies worldwide with poor prognosis for lack of early detection and effective treatment modalities. The significant influence of tumor microenvironment on malignant cells has been extensively investigated in this targeted-therapy era. Epithelial-mesenchymal transition (EMT) is a highly conserved and fundamental process that is critical for embryogenesis and some other pathophysiological processes, especially tumor genesis and progression. Aberrant gastric EMT activation could endow gastric epithelial cells with increased mesenchymal characteristics and less epithelial features, and promote cancer cell stemness, initiation, invasion, metastasis, and chemo-resistance with cellular adhesion molecules especially E-cadherin concomitantly repressed, which allows tumor cells to disseminate and spread throughout the body. Some pathogens, stress, and hypoxia could induce and aggravate GC via EMT, which is significantly correlated with prognosis. GC EMT is modulated by diverse micro-environmental, membrane, and intracellular cues, and could be triggered by various overexpressed transcription factors, which are downstream of several vital cross-talking signaling pathways including TGF-β, Wnt/β-catenin, Notch, etc. microRNAs also contribute significantly to GC EMT modulation. There are currently some agents which could suppress GC EMT, shedding light on novel anti-malignancy strategies. Investigating potential mechanisms modulating GC cell EMT and discovering novel EMT regulators will further elucidate GC biology, and may provide new biomarkers for early GC detection and potentially efficient targets for preventative and curative anti-GC intervention approaches to prevent local and distant invasions.

Objective: Mucinous cystadenocarcinoma of appendix is a rare entity. Differentiating mucinous cystadenocarcinoma from mucinous cystadenoma is very challenging and depends on establishing the presence of malignant cells in the appendix wall. The invasion may be very difficult to assess in some cases, especially in early stages of the disease, which could have devastating prognostic effects on patients. Therefore, it is necessary to develop an ancillary test that can differentiate the mucinous cystadenocarcinoma from mucinous cystadenoma. So far, there is no report available about the role of differentially expressed miRNAs in the diagnosis of appendiceal mucinous cystadenocarcinoma., Materials and Methods: Six confirmed mucinous appendiceal cystadenocarcinoma and twelve mucinous appendiceal cystadenoma cases were selected. The total RNAs were extracted from the formalin-fixed paraffin-embedded specimen of these cases. The comprehensive miRNA microarray expression profiling from pooled aliquots of RNA samples from these two entities were analyzed to detect the differentially expressed miRNAs in mucinous cystadenocarcinoma. The best seven differentially expressed miRNAs were validated in individual cases by quantitative reverse transcriptase PCR (qRT-PCR)., Results: The microarray miRNA expression profiling analysis revealed 646 miRNAs that were differentially expressed in the mucinous cystadenocarcinoma. Among these differentially expressed miRNAs, the expression of 80 miRNAs showed statistical difference (p<0.01). The quantitative RT-PCR validated that the expression of miR-1, miR-4328 was significantly down regulated in mucinous cystadenocarcinoma compared to the mucinous cystadenoma (p<0.05). On the other hand, the expression of miR-200b, miR-200c, miR-451, miR-223 and miR-21 were significantly upregulated in mucinous cystadenocarcinoma (p<0.05)., Conclusion: The expression levels of miRNAs tested were significantly altered in the appendiceal mucinous cystadenocarcinoma samples compared to the mucinous cystadenoma. These data suggest that the miRNA expression in mucinous appendiceal neoplasm may help to supplement the morphological evaluation in distinguishing benign from malignant tumors.

Objective: There is an increased risk of developing ovarian cancer (OC) in patients with endometriosis. Hence, development of new biomarkers may provide a positive clinical outcome for early detection. MicroRNAs (miRNAs) are small non-coding RNAs that play an important role in biological and pathological process and are currently used as diagnostic and prognostic markers in various cancers. In the current study, we assessed the differential expression of miRNAs from 19 paired ovarian cancer and its associated endometriosis tissue samples. In addition we also analyzed the downstream targets of those miRNAs., Methods: Nineteen paired cases of ovarian cancer and endometriosis foci were identified by a gynecologic pathologist and macro-dissected. The total RNAs were extracted and subjected to comprehensive miRNA profiling from the pooled samples of these two different entities using microarray analysis. Later, the abnormal expressions of few selected miRNAs were validated in individual cases by quantitative real-time PCR (qRT-PCR). Ingenuity pathway analysis revealed target mRNAs which were validated by qRT-PCR., Results: The miRNA profiling identified deregulation of greater than 1156 miRNAs in OC, of which the top seven were further validated by qRT-PCR. The expression of miR-1 , miR-133a , and miR-451 were reduced significantly (p<0.0001) in the OC patients compared to its associated endometriosis. In contrast, the expression of miR-141 , miR-200a , miR-200c , and miR-3613 were elevated significantly (p<0.05) in most of the OC patients. Furthermore, among the downstream mRNAs of these miRNAs, the level of PTEN expression was significantly (p<0.05) reduced in OC compared to endometriosis while no significant difference was observed in NF-κB expression., Conclusion: The expression of miRNAs and mRNAs in OC were significantly different compared to its concurrent endometriosis. These differential expressed miRNAs may serve as potential diagnostic and prognostic biomarkers for OC associated with endometriosis.

In order to study the influencing factors on Raman spectroscopy, we research a series of comparative Raman spectroscopy of multi-walled carbon nanotubes (MWCNT) with different tube diameter and length. The results suggest that the G peak and D peak of MWCNT are all red-shifted as compared to that of polycrystalline graphite; In the same conditions, the peak intensity (G peak and D peak) is directly proportional to the diameter of the MWCNT, and inversely proportional to the length of the MWCNT; G peak frequency shift is closely related to the MWCNT diameter and length, which are inversely proportional to the diameter (with identical results of the single-walled carbon nanotube radial breathing modes) and direct proportional to the length. While, the influences of the diameter and length on D peak frequency shift are weak, and future analysis for the reason of this kind of phenomenon is as follows. Subsequently, we investigated the relation between D peak frequency shift and MWCNT aspect ratio, the relationship between G peak frequency shift and aspect ratio is nearly linear increase. Using the same analysis method, we plotted the different graphs of G peak and D peak intensity vs the aspect ratio of MWCNT, respectively. As the expected, the linear degression relation are existent in the two relationships.

Premise of the Study: Microsatellite loci were developed for Prunus sibirica to investigate genetic diversity, population genetic structure, and marker-assisted selection of late-blooming cultivars in the breeding of P. sibirica. •, Methods and Results: Using a magnetic bead enrichment strategy, 19 primer pairs were developed and characterized across 40 individuals from three P. sibirica wild populations and six individuals of P. armeniaca. The number of alleles per locus varied from three to 11 and the observed and expected heterozygosities ranged from 0.063 to 0.917 and 0.295 to 0.876, respectively, in the three P. sibirica wild populations. All primer pairs could be successfully amplified in six individuals of P. armeniaca. •, Conclusions: These microsatellite primer pairs should be useful for population genetics, germplasm identification, and marker-assisted selection in the breeding of P. sibirica and related species.

Background: Errors that increase the risk of wrong-side/-site procedures not only occur the day of surgery but also are often introduced much earlier during the scheduling process. The frequency of these booking errors and their effects are unclear., Methods: All surgical scheduling errors reported in the institution's medical event reporting system from January 1, 2011, to July 31, 2011, were analyzed. Focus groups with operating room nurses were held to discuss delays caused by scheduling errors., Results: Of 17,606 surgeries, there were 151 (.86%) booking errors. The most common errors were wrong side (55, 36%), incomplete (38, 25%), and wrong approach (25, 17%). Focus group participants said incomplete and wrong-approach bookings resulted in the longest delays, averaging 20 minutes and costing at least $320., Conclusions: Although infrequent, scheduling errors disrupt operating room team dynamics, causing delays and bearing substantial costs. Further research is necessary to develop tools for more accurate scheduling., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Treacher Collins syndrome (TCS) is a craniofacial syndrome that is both phenotypically variable and heterogeneous, caused by mutations in the TCOF1, POLR1C, and POLR1D genes. We examined attitudes towards TCS prenatal genetic testing among affected families using a telephone questionnaire. Participants were 31 affected adults and relatives recruited primarily through families cared for in the mid-Atlantic region. Nineteen participants (65%) reported that they would take a TCS prenatal genetic test which could not predict degree of disease severity. Interest in TCS genetic testing was associated with higher income, higher concern about having a child with TCS, lower religiosity, lower concern about genetic testing procedures, and having a sporadic rather than familial mutation. Over half reported that their decision to have TCS genetic testing would be influenced a great deal by their desire to relieve anxiety and attitudes toward abortion. Ten participants (32%) reported that they would be likely to end the pregnancy upon receiving a positive test result; this was lower amongst TCS affected individuals and higher amongst participants with children with TCS. Genetics healthcare providers need to be aware of affected individuals' and families' attitudes and interest in prenatal genetic testing for TCS, and the possible implications for other craniofacial disorders, so that patients' information needs can be met., (Copyright © 2012 Wiley Periodicals, Inc.)

A structure-activity relationship study was undertaken to address the lack of oral exposure of the H3 antagonist 1, which incorporated an arylketone. Among a number of sub-series, the 4H-pyrido[1,2-a]pyrimidin-4-one analog 21 showed an improved PK profile in rat and mouse and was active in an obesity model. The pyrimidin-4-one proved to be a novel and useful ketone bioisostere., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The synthesis and SAR of a novel series of 4-azabenzoxazole histamine H(3) antagonists is described. Introduction of substituted phenyl, pyridyl and fused heterocyclic groups to the 6-position of the 4-azabenzoxazole core gave a series of compounds with good H(3) antagonist activity in both ex vivo and in vivo assays., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

A series of novel 2-piperidinopiperidine thiadiazoles were synthesized and evaluated as new leads of histamine H3 receptor antagonists. The 4-(5-([1,4'-bipiperidin]-1'-yl)-1,3,4-thiadiazol-2-yl)-2-(pyridin-2-yl)morpholine (5u) displayed excellent potency and ex vivo receptor occupancy. Compound 5u was also evaluated in vivo for antidiabetic efficacy in STZ diet-induced obesity type 2 diabetic mice for 2 or 12 days. Non-fasting glucose levels were significantly reduced as compared with vehicle-treated mice. In addition, 5u dose dependently blocked the increase of HbA1c after 12 days of treatment.

Background: Patient-reported outcomes in cleft lip and palate treatment are critical for patient care. Traditional surgical outcomes focused on objective measures, such as photographs, anatomic measurements, morbidity, and mortality. Although these remain important, they leave many questions unanswered. Surveys that include aesthetics, speech, functionality, self-image, and quality of life provide more thorough outcomes assessment. It is vital that reliable, valid, and comprehensive questionnaires are available to craniofacial surgeons., Methods: The authors performed a literature review to identify questionnaires validated in cleft lip and palate patients. Qualifying instruments were assessed for adherence to guidelines for development and validation by the scientific advisory committee and for content., Results: The authors identified 44 measures used in cleft lip and palate studies. After 15 ad hoc questionnaires, eight generic instruments, 11 psychiatric instruments, and one non-English language questionnaire were excluded, nine measures remained. Of these, four were never validated in the cleft population. Analysis revealed one craniofacial-specific measure (Youth Quality of Life-Facial Differences), two voice-related measures (Patient Voice-Related Quality of Life and Cleft Audit Protocol for Speech-Augmented), and two oral health-related measures (Child Oral Health Impact Profile and Child Oral Health Quality of Life). The Youth Quality of Life-Facial Differences, Child Oral Health Impact Profile, and Child Oral Health Quality of Life questionnaires were sufficiently validated. None was created specifically for clefts, resulting in content limitations., Conclusions: There is a lack of comprehensive, valid, and reliable questionnaires for cleft lip and palate surgery. For thorough assessment of satisfaction, further research to develop and validate cleft lip and palate surgery-specific instruments is needed.

Great progress has been made in genetic dissection of quantitative trait variation during the past two decades, but many studies still reveal only a small fraction of quantitative trait loci (QTLs), and epistasis remains elusive. We integrate contemporary knowledge of signal transduction pathways with principles of quantitative and population genetics to characterize genetic networks underlying complex traits, using a model founded upon one-way functional dependency of downstream genes on upstream regulators (the principle of hierarchy) and mutual functional dependency among related genes (functional genetic units, FGU). Both simulated and real data suggest that complementary epistasis contributes greatly to quantitative trait variation, and obscures the phenotypic effects of many 'downstream' loci in pathways. The mathematical relationships between the main effects and epistatic effects of genes acting at different levels of signaling pathways were established using the quantitative and population genetic parameters. Both loss of function and "co-adapted" gene complexes formed by multiple alleles with differentiated functions (effects) are predicted to be frequent types of allelic diversity at loci that contribute to the genetic variation of complex traits in populations. Downstream FGUs appear to be more vulnerable to loss of function than their upstream regulators, but this vulnerability is apparently compensated by different FGUs of similar functions. Other predictions from the model may account for puzzling results regarding responses to selection, genotype by environment interaction, and the genetic basis of heterosis.

A series of 2-piperidinopiperidine-5-arylthiadiazoles was synthesized and subjected to a structure-activity relationship (SAR) investigation. The potency of this series was improved to the single digit nanomolar range. The key analogs were shown to be free of P450 issues, and they also maintained good ex vivo activity and brain penetration., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels mediate the hyperpolarization-activated current I(h) and thus play important roles in the regulation of brain excitability. The subcellular distribution pattern of the HCN channels influences the effects that they exert on the properties and activity of neurons. However, little is known about the mechanisms that control HCN channel trafficking to subcellular compartments or that regulate their surface expression. Here we studied the dynamics of HCN channel trafficking in hippocampal neurons using dissociated cultures coupled with time lapse imaging of fluorophore-fused HCN channels. HCN1-green fluorescence protein (HCN1-GFP) channels resided in vesicle-like organelles that moved in distinct patterns along neuronal dendrites, and these properties were isoform-specific. HCN1 trafficking required intact actin and tubulin and was rapidly inhibited by activation of either NMDA or AMPA-type ionotropic glutamate receptors in a calcium-dependent manner. Glutamate-induced inhibition of the movement of HCN1-GFP-expressing puncta was associated with increased surface expression of both native and transfected HCN1 channels, and this surface expression was accompanied by augmented I(h). Taken together, the results reveal the highly dynamic nature of HCN1 channel trafficking in hippocampal neurons and provide a novel potential mechanism for rapid regulation of I(h), and hence of neuronal properties, via alterations of HCN1 trafficking and surface expression.

A series of 2-(1,4'-bipiperidine-1'-yl)thiazolopyridines was synthesized and evaluated as a new lead of non-imidazole histamine H(3) receptor antagonists. Introduction of diversity at the 6-position of the pyridine ring was designed to enhance in vitro potency and decrease hERG activity. The structure-activity relationships for these new thiazolopyridine antagonists are discussed.