Search

Alkaline thermal hydrolysis of sewage sludge produces nutrients and biostimulants that enhance plant growth, attracting considerable interest in agriculture. However, the metabolic differences and regulatory mechanisms of sewage sludge-derived biostimulants (SS-BS) on the phenotypic traits, nutritional quality, and safety indicators of harvested crops remain unclear. This study investigates the impact of SS-BS on rice quality on an agricultural production scale. The research reveals that rice treated with SS-BS complies with safety standards comparable to premium rice. SS-BS significantly enhances nutrient enrichment in the endosperm, increasing protein, vitamin B1, dietary fiber, and vitamin E content by 7%, 7.2%, 23.2%, and 42.2%, respectively. Furthermore SS-BS upregulates the FG2 gene,leading to increased Nictoflorin content and activation of the gene expression of UGT73C6 and CYP75A, which catalyze O-glycosylation and promot glycosyl transfer. By inhibiting the synthesis of Trifolin, Scolymoside, and Swertiajaponin, SS-BS favors the synthesis of glycosylated derivatives of Tricin and Luteolin, which exhibit higher anti-inflammatory activity. Additionally, two novel genes, novel.2100 and novel.1300, and an uncharacterized gene, LOC9269295, are closely associated with the production of anti-inflammatory and antioxidant compounds. This study provides new evidence for SS-BS application and insights into their regulatory mechanisms affecting crop quality, contributing to the development of functional foods and sustainable agriculture., 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 © 2024 Elsevier Inc. All rights reserved.)

Limited scientific evidence exists on phosphorus immobilization under autogenetic electrochemical reactions in composting systems. This study exploited a composting procedure using microbial fuel cell (MFC) to ascertain phosphorus redistribution during composting process. Compared to the control without MFC equipment, MFC-assisted treatment yielded a 13 % decrease in phosphorus availability due to the transformation of exchangeable fraction (Ex-P) to aluminum-bound (Al-P) and calcium-bound (Ca-P) fractions. During the composting process, organic humification primarily controlled phosphorus redistribution and immobilization. Biotic factors, including bacterial communities (i.e., Firmicutes, Proteobacteria, Bacteroidota, and Gemmatimonadota) and functional enzymes (i.e., acid phosphatase, alkaline phosphatase, phytase, and C-P lyase), significantly influenced phosphorus availability in the composting systems. Temperature-dependent composting phases restricted microbial actions on phosphorus transformation. These findings highlight the mechanisms underlying phosphorus transformation in composting systems, and provide valuable insights for advancing composting technology and protecting agricultural ecosystems., 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 © 2024 Elsevier Ltd. All rights reserved.)

Little scientific evidence on metal passivation, antibiotic degradation and resistance genes removal, is available under autogenetic electrochemical reactions during composting process. This study established microbial fuel cell (MFC)-assisted composting procedure to ascertain the removal performance and detoxification mechanisms involving metals, antibiotics and their resistance genes. Compared to control treatment, the bioavailability of zinc (Zn) and copper (Cu) in MFC-assisted treatment decreased by 7.8% and 26.9%, while the content of tetracycline (TCL) and oxytetracycline (OCL) reduced by 100% and 89%, respectively. Organics mineralization and humification were responsible for 80% and 70% of the variations in metal passivation and antibiotic degradation during composting process. A decrease of 54% was found for tetW gene, while copA gene increased by 42% in MFC-assisted composting treatment. These findings highlight the detoxification mechanisms underlying metal passivation and antibiotic degradation during composting process, and potentially offer valuable insights for environmental source protection and agricultural sustainable development., 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 © 2024 Elsevier Inc. All rights reserved.)

Background: The post COVID-19 health condition of Chinese residents infected with Omicron is not clear after the change of epidemic prevention policies. This study aimed to clarify the epidemiology and associated factors about health status of rehabilitation patients., Methods: A quick questionnaire study based on C19-YRSm was conducted in mainland China through internet from May 1, 2023, to May 7, 2023. Chinese native speakers infected with Omicron variant agreed to participate were included. Persisting symptom and living habits were simultaneously inquired. Logistic regression analysis was used to identify the associated factors., Results: In this study 753 individuals were included. Of whom 57.90% were males, 89.38% did not seek medical service, 99.47% recovered within less than 120 days. Breathlessness (47.68%), cognitive impairment (44.89%), Anxiety/mood changes (33.20%), pain/discomfort (32.94%), fatigue or tiredness not improved by rest (32.27%) and post-exertional malaise (30.01%) were the top reported key symptoms. Less than 10% respondents reported functional limitations. The prevalence of fever was reported greater than that of other symptoms, with dry eyes at 14.87%, appetite change at 14.34%, and hair loss at 12.22%. Middle age (OR: 2.353, 95%CI: 1.171 ~ 4.729), underlying diseases (OR: 2.293, 95%CI: 1.216 ~ 4.324), severe key symptom (OR: 6.168, 95%CI: 1.376 ~ 27.642) and at least one other symptom (OR: 1.847, 95%CI: 1.225 ~ 2.718)during the recovery were the risk factors of poor overall health after infection (current overall health score <8; 74.10%), while daily exercise in recovery period (OR: 0.457, 95%CI: 0.229 ~ 0.913), a low-fat diet (OR: 0.600, 95%CI: 0.401 ~ 0.898) and the recovery time from 2 to 4 months (OR: 0.639, 95%CI: 0.445 ~ 0.918) were the protective factors., Conclusion: This is the first time to use the C19-YRSm scale to evaluate the health status in China. The study revealed prevalence of persistent symptoms within 120 days after Omicron onset., (© 2024. The Author(s).)

Bone age assessment (BAA) is crucial in various fields, including legal proceedings, athletic competitions, and clinical medicine. However, the use of X-ray methods for age estimation without medical indication is subject to ethical debate, especially in forensic and athletic fields. The application of magnetic resonance imaging (MRI) with non-ionizing radiation can overcome this limitation in BAA. This study aimed to compare the application value of several MRI modalities of proximal humeral in BAA. A total of 468 patients with shoulder MRIs were retrospectively collected from a Chinese Han population aged 12-30 years (259 males and 209 females) for training and testing, including T 1 weighted MRI (T 1 WI), T 2 weighted MRI (T 2 WI), and Proton density weighted MRI (PDWI). Optimal regression models were established for age estimation, yielding mean absolute error (MAE) values below 2.0 years. The MAE values of T 1 WI were the lowest, with 1.700 years in males and 1.798 years in females. The area under the curve (AUC) and accuracy values of different MRI modalities of 16-year and 18-year thresholds were all around 0.9. For the 18-year threshold, T 1 WI outperformed T 2 WI and PDWI. In conclusion, the three MRI modalities of the proximal humerus can serve as reliable indicators for age assessment, while the T 1 WI performed better in age assessment and classification., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Cryptococcal meningitis (CM) is a fatal fungal central nervous system (CNS) infection caused by Cryptococcus infecting the meninges and/or brain parenchyma, with fever, headache, neck stiffness, and visual disturbances as the primary clinical manifestations. Immunocompromised individuals with human immunodeficiency virus (HIV) infection or who have undergone organ transplantation, as well as immunocompetent people can both be susceptible to CM. Without treatment, patients with CM may have a mortality rate of up to 100% after hospital admission. Even after receiving therapy, CM patients may still suffer from problems such as difficulty to cure, poor prognosis, and high mortality. Therefore, timely and effective treatment is essential to improve the mortality and prognosis of CM patients. Currently, the clinical outcomes of CM are frequently unsatisfactory due to limited drug choices, severe adverse reactions, drug resistance, etc. Here, we review the research progress of CM treatment strategies and discuss the suitable options for managing CM, hoping to provide a reference for physicians to select the most appropriate treatment regimens for CM patients., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany.)

Background: COVID-19 caused by SARS-CoV-2 is a great threat to public health. We present the safety and immunogenicity data from a phase I trial in China of an mRNA vaccine (LVRNA009)., Methods: In the single-centre, double-blind, placebo-controlled and dose-escalation study, 72 healthy unvaccinated adults aged 18-59 years were randomized (3:1) to receive LVRNA009 with one of three vaccine dosage (25, 50 and 100 μg) or placebo, to evaluate for the safety, tolerability and immunogenicity of LVRNA009., Results: All these participants received two injections 28 days apart. No adverse events higher than grade 2 were reported during the study. A total of 30 participants (42 %) reported solicited adverse reactions during the first 14 days after vaccinations. Of the events reported, fever (n = 11, 15 %) was the most common systemic adverse reaction, and pain at the injection site (n = 17, 24 %) was the most frequent solicited local adverse reaction. Anti-S-protein IgG and neutralising antibodies were observed to have been induced 14 days after the first dose, significantly increased 7 days after the second dose, and remained at a high level 28 days after the second dose. Specific T-cell responses peaked 7 days and persisted 28 days after second vaccination., Conclusion: LVRNA009 has demonstrated promising results in safety and tolerability at all three dose levels among Chinese adults. LVRNA009 at three dose levels could rapidly induce strong humoral and cellular immune responses, including binding and neutralising antibody production and IFN- γ secretion, which showed good immunogenicity., Clinical Trial Registration Number: Clinicaltrials.gov NCT05364047; Chictr.org.cn ChiCTR2100049349., 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 © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Biomass-derived carbonaceous materials with graphene/graphene-like structures (BGS) have attracted tremendous attention in the field of environmental remediation. The introduction of graphene/graphene-like structures into raw biochars can effectively improve their properties, such as electrical conductivity, surface functional groups, and catalytic activity. In 2021, the International Organization for Standardization defined graphene as a "single layer of carbon atoms with each atom bound to three neighbours in a honeycomb structure". Considering this definition, several studies have incorrectly referred to BGS (e.g., biomass-derived few-layer graphene or porous graphene-like nanosheets) as "graphene". The definitions and classifications of BGS and their applications in environmental remediation have not been assessed critically thus far. Comprehensive analysis and sufficient and robust evidence are highly desired to accurately determine the specific structures of BGS. In this perspective, we provide a systematic framework to define and classify the BGS. The state-of-the-art methods currently used to determine the structural properties of BGS are scrutinized. We then discuss the design and fabrication of BGS and how their distinctive features could improve the applicability of biomass-derived carbonaceous materials, particularly in environmental remediation. The environmental applications of these BGS are highlighted, and future research opportunities and needs are identified. The fundamental insights in this perspective provide critical guidance for the further development of BGS for a wide range of environmental applications.

Objective: To investigate the effects of acupuncture on JNK pathway and autophagy level in rats with intracerebral hemorrhage （ICH） and explore the partial mechanism of acupuncture against ICH., Methods: SD rats were randomly divided into blank group, model group and acupuncture group. Each group was divided into Day 1, Day 3 and Day 7 subgroups respectively, with 5 rats in each group. The autologous blood injection was adopted to duplicate rat model of ICH. In the acupuncture group, the needle was inserted from "Baihui" （GV20） towards "Qubin" （GB7） on the affected side, stimulating for 30 min each time, once daily； the same acupuncture technique was opera-ted in each subgroup for 1, 3 and 7 days, separately. Using Bederson scale, the neurological deficit was evaluated in each group. Western blot was adopted to detect the protein expression levels of Beclin1, LC3Ⅰ/Ⅱ, phosphorylated c-Jun amino-terminal kinase （p-JNK） and the phosphorylated （p）-c-Jun around hematoma lesion of the brain tissue of rats in each group., Results: After treatment, the neurological deficit score of rats in the model group was higher than that of the blank group at each time point （ P <0.05）, and the score of the acupuncture group started declining since the 3 rd day of treatment when compared with the model group （ P <0.05）. At each time point, compared with the blank group, the protein expression levels of LC3Ⅰ/Ⅱ, Beclin1, p-c-Jun and p-JNK was increased （ P <0.01）. Compared with the model group, the protein expression level of LC3Ⅰ/Ⅱ was reduced （ P <0.05）； the protein expression levels of Beclin1, p-c-Jun and p-JNK was increased （ P <0.05, P <0.01） on day 3 and 7 in the acupuncture group., Conclusion: Acupuncture can activate the JNK pathway in the brain tissue of rats with ICH and increase the level of autophagy, thereby improving the neurological function of the rats with ICH.

Background: The genus Cantonius Théry, 1929 is a small group with two subgenera and 12 species. However, the biology of this genus is still unknown., New Information: In this paper, three species of the genus Cantonius Théry, 1929 were found on bamboo leaves, revealing for the first time that Cantonius species are also leaf-miners. Two new species were recorded from Jiangxi Province and are described here: Cantonius (Cantonius) anjiensis sp. n. (host plant: Pleioblastusamarus ) and Cantonius (Procantonius) qiyunensis sp. n. (host plant: Bambusablumeana ) followed by C. (P.) austrisinicus Kalashian, 2021 (host plant: Oligostachyumpaniculatum ) recorded from Guangxi Province. Including habitats, photos of three species together with C. (P.) qiyunensis sp. n. pupa, host plants, and leaf mines of the three species are presented. Moreover, the bionomics and habits of the genus are discussed for the first time, and a hypothesis for the distribution of Cantonius is provided., (Xiao-Di Shi, Zhi-Ming Wu, Xiao-Hua Dai, Jia-Sheng Xu, Hai-Tian Song.)

Bundle sheath cells play a crucial role in photosynthesis in C4 plants, but the structure and function of photosystem II (PSII) in these cells is still controversial. Photoprotective roles of bundle sheath chloroplasts at the occurrence of environmental stresses have not been investigated so far. Non-photochemical quenching (NPQ) of chlorophyll a fluorescence is the photoprotective mechanism that responds to a changing energy balance in chloroplasts. In the present study, we found a much higher NPQ in bundle sheath chloroplasts than in mesophyll chloroplasts under a drought stress. This change was accompanied by a more rapid dephosphorylation of light-harvesting complex II (LHCII) subunits and a greater increase in PSII subunit S (PsbS) protein abundance than in mesophyll cell chloroplasts. Histochemical staining of reactive oxygen species (ROS) suggested that the high NPQ may be one of the main reasons for the lower accumulation of ROS in bundle sheath chloroplasts. This may maintain the stable functioning of bundle sheath cells under drought condition. These results indicate that the superior capacity for dissipation of excitation energy in bundle sheath chloroplasts may be an environmental adaptation unique to C4 plants., 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 Liu, Liu, Chen, Yin, Zhang, Song, Chang, Zhang, Wang, Dai, Wei, Xiong, Yuan and Zhao.)

Uterine receptivity to the embryo is crucial for successful implantation. The establishment of uterine receptivity requires a large amount of energy, and abnormal energy regulation causes implantation failure. Glucose metabolism in the endometrium is tissue specific. Glucose is largely stored in the form of glycogen, which is the main energy source for the endometrium. AMP-activated protein kinase (AMPK), an important energy-sensing molecule, is a key player in the regulation of glucose metabolism and its regulation is also tissue specific. However, the mechanism of energy regulation in the endometrium for the establishment of uterine receptivity remains to be elucidated. In this study, we aimed to investigate the energy regulation mechanism of mouse uterine receptivity and its significance in embryo implantation. The results showed that the AMPK, p-AMPK, glycogen synthase 1, and glycogen phosphorylase M levels and the glycogen content in mouse endometrial epithelium varied in a periodic manner under regulation by the ovarian hormone. Specifically, progesterone significantly activated AMPK, promoted glycogenolysis, and upregulated glycogen phosphorylase M expression. AMPK regulated glycogen phosphorylase M expression and promoted glycogenolysis. AMPK was also found to be activated by changes in the energy or glycogen of the endometrial epithelial cells. The inhibition of AMPK activity or glycogenolysis altered the uterine receptivity markers during the window of implantation and ultimately interfered with implantation. In summary, consistency and synchronization of AMPK and glycogen metabolism constitute the core regulatory mechanism in mouse endometrial epithelial cells involved in the establishment of uterine receptivity.

BACKGROUND The aims of the study were to comprehensively compare the morphology, immunophenotype, proliferation, migration, and regeneration potential of normal dental pulp stem cells (DPSCs) versus inflammatory dental pulp stem cells (iDPSCs). MATERIAL AND METHODS Healthy pulp or inflamed pulp tissue was used to isolate and culture DPSCs and iDPSCs, respectively. These cell populations were characterized by flow cytometry, colony formation assay, transwell assay, and multi-directional differentiation in vitro. RESULTS No difference was observed in the morphology, cell-surface markers, or cell migration between DPSCs and iDPSCs. DPSCs showed a higher colony-forming capacity, proliferative viability, and osteo/dentinogenesis ability compared with iDPSCs. However, iDPSCs demonstrated enhanced neurogenesis, angiogenesis, adipogenesis, and chondrogenesis capacities in comparison to DPSCs. CONCLUSIONS Our data revealed the differences of biological properties between DPSCs and iDPSCs. The highly angiogenic and neurogenic potential of iDPSCs indicate their possible use in the regeneration of the dentin-pulp complex and support the critical role of angiogenesis and neurogenesis in pulp regeneration.

Background: Safe and effective vaccines are urgently needed to end the COVID-19 pandemic caused by SARS-CoV-2 infection. We aimed to assess the preliminary safety, tolerability, and immunogenicity of an mRNA vaccine ARCoV, which encodes the SARS-CoV-2 spike protein receptor-binding domain (RBD)., Methods: This single centre, double-blind, randomised, placebo-controlled, dose-escalation, phase 1 trial of ARCoV was conducted at Shulan (Hangzhou) hospital in Hangzhou, Zhejiang province, China. Healthy adults aged 18-59 years negative for SARS-CoV-2 infection were enrolled and randomly assigned using block randomisation to receive an intramuscular injection of vaccine or placebo. Vaccine doses were 5 μg, 10 μg, 15 μg, 20 μg, and 25 μg. The first six participants in each block were sentinels and along with the remaining 18 participants, were randomly assigned to groups (5:1). In block 1 sentinels were given the lowest vaccine dose and after a 4-day observation with confirmed safety analyses, the remaining 18 participants in the same dose group proceeded and sentinels in block 2 were given their first administration on a two-dose schedule, 28 days apart. All participants, investigators, and staff doing laboratory analyses were masked to treatment allocation. Humoral responses were assessed by measuring anti-SARS-CoV-2 RBD IgG using a standardised ELISA and neutralising antibodies using pseudovirus-based and live SARS-CoV-2 neutralisation assays. SARS-CoV-2 RBD-specific T-cell responses, including IFN-γ and IL-2 production, were assessed using an enzyme-linked immunospot (ELISpot) assay. The primary outcome for safety was incidence of adverse events or adverse reactions within 60 min, and at days 7, 14, and 28 after each vaccine dose. The secondary safety outcome was abnormal changes detected by laboratory tests at days 1, 4, 7, and 28 after each vaccine dose. For immunogenicity, the secondary outcome was humoral immune responses: titres of neutralising antibodies to live SARS-CoV-2, neutralising antibodies to pseudovirus, and RBD-specific IgG at baseline and 28 days after first vaccination and at days 7, 15, and 28 after second vaccination. The exploratory outcome was SARS-CoV-2-specific T-cell responses at 7 days after the first vaccination and at days 7 and 15 after the second vaccination. This trial is registered with www.chictr.org.cn (ChiCTR2000039212)., Findings: Between Oct 30 and Dec 2, 2020, 230 individuals were screened and 120 eligible participants were randomly assigned to receive five-dose levels of ARCoV or a placebo (20 per group). All participants received the first vaccination and 118 received the second dose. No serious adverse events were reported within 56 days after vaccination and the majority of adverse events were mild or moderate. Fever was the most common systemic adverse reaction (one [5%] of 20 in the 5 μg group, 13 [65%] of 20 in the 10 μg group, 17 [85%] of 20 in the 15 μg group, 19 [95%] of 20 in the 20 μg group, 16 [100%] of 16 in the 25 μg group; p<0·0001). The incidence of grade 3 systemic adverse events were none (0%) of 20 in the 5 μg group, three (15%) of 20 in the 10 μg group, six (30%) of 20 in the 15 μg group, seven (35%) of 20 in the 20 μg group, five (31%) of 16 in the 25 μg group, and none (0%) of 20 in the placebo group (p=0·0013). As expected, the majority of fever resolved in the first 2 days after vaccination for all groups. The incidence of solicited systemic adverse events was similar after administration of ARCoV as a first or second vaccination. Humoral immune responses including anti-RBD IgG and neutralising antibodies increased significantly 7 days after the second dose and peaked between 14 and 28 days thereafter. Specific T-cell response peaked between 7 and 14 days after full vaccination. 15 μg induced the highest titre of neutralising antibodies, which was about twofold more than the antibody titre of convalescent patients with COVID-19., Interpretation: ARCoV was safe and well tolerated at all five doses. The acceptable safety profile, together with the induction of strong humoral and cellular immune responses, support further clinical testing of ARCoV at a large scale., Funding: National Key Research and Development Project of China, Academy of Medical Sciences China, National Natural Science Foundation China, and Chinese Academy of Medical Sciences., Competing Interests: C-FQ and BY are co-inventors on pending patent applications related to the ARCoV mRNA vaccine. LW is an employee of Suzhou Abogen Biosciences. S-YY and ZH are employees of Walvax. All other authors declare no competing interests., (© 2022 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY-NC-ND 4.0 license.)

Intracerebral haemorrhage (ICH) can be a catastrophic event; even if the initial stages of the pathology were well-managed, a number of patients experience varied residual neurological deficits following the insult. Ferroptosis is a recently identified type of cell demise which is tightly linked to the neurological impairment associated with ICH. In the current work, the prophylactic impact of scalp acupuncture (SA) therapy on autologous blood injection murine models of ICH was investigated in order to establish whether SA could mitigate the secondary damage arising following ICH by moderating ferroptosis. The pathophysiological mechanisms associated with this process were also explored. Ludmila Belayev tests were utilised for the characterisation of neurological damage. Haematoxylin-eosin staining was employed in order to determine the cerebral impact of the induced ICH. Malondialdehyde (MDA) and iron titres in peri-haemorrhagic cerebral tissues were appraised using purchased assay kits. Transmission electron microscopy delineated mitochondrial appearances within nerve cell bodies from the area of haemorrhage. Western blotting techniques were utilised to assay the degree of protein expression of NeuN, sequestosome 1 (p62), nuclear factor erythroid 2-related factor 2 (Nrf2), Kelch-like ECH-associated protein 1 (Keap1), glutathione peroxidase 4 (GPX4) and ferritin heavy chain 1 (FTH1). The frequencies of Nrf2, GPX4 and FTH1 positive cells, respectively, were documented with immunohistochemical staining. The results demonstrated that therapy with SA after ICH mitigated MDA and iron sequestration, diminished the appearance of contracted mitochondria with increased outer mitochondrial membrane diameter within the nerve cell bodies, and suppressed neuronal ferroptosis. The pathways responsible for these effects may encompass amplified p62, Nrf2, GPX4 and FTH1 expression, together with decreased Keap1 expression. Application of SA reduced identified neurobehavioural abnormalities after ICH; no disparities were observed between the consequences of SA therapy and deferoxamine delivery. It can be surmised that intervention with SA enhanced recovery after ICH by triggering the antioxidant pathway, p62/Keap1/Nrf2, and causing FTH1 and GPX4 upregulation, factors that participate in diminishing excess iron and thus in mitigating lipid peroxidation insults arising from ferroptosis following ICH., (© 2021. The Author(s).)

Objective: To explore the fluid resuscitation strategy in shock stage in severely burned children with different burn areas in different age groups, and to evaluate the curative effect. Methods: A retrospective cohort study was conducted. From January 2015 to June 2020, 235 children with severe and above burns who met the inclusion criteria were hospitalized in the First Affiliated Hospital of Nanchang University, including 150 males and 85 females, aged 3 months to 12 years. After admission, it was planned to rehydrate the children with electrolyte, colloid, and water according to the domestic rehydration formula for pediatric burn shock, and the rehydration volume and speed were adjusted according to the children's mental state, peripheral circulation, heart rate, blood pressure, and urine output, etc. The actual input volume and planned input volume of electrolyte, colloid, water, and total fluid of all the children were recorded during the 8 hours since fluid replacement and the first and second 24 hours after injury. According to urine output during the 8 hours since fluid replacement, all the children were divided into satisfactory urine output maintenance group (119 cases) with urine output ≥1 mL·kg -1 ·h -1 and unsatisfactory urine output maintenance group (116 cases) with urine output <1 mL·kg -1 ·h -1 , and the electrolyte coefficient, colloid coefficient, and water coefficient of the children were calculated during the 8 hours since fluid replacement. According to the total burn area, children aged <3 years (155 cases) and 3-12 years (80 cases) were divided into 15%-25% total body surface area (TBSA) group and >25%TBSA group, respectively. The electrolyte coefficient, colloid coefficient, water coefficient, and urine output of the children were calculated or counted during the first and second 24 hours after injury, and the non-invasive monitoring indicators of body temperature, heart rate, respiratory rate, and percutaneous arterial oxygen saturation and efficacy indicators of hematocrit, platelet count, hemoglobin, albumin, creatinine, and alanine aminotransferase (ALT) of the children were recorded 48 hours after injury. The prognosis and outcome indicators of all the children during the treatment were counted, including complications, cure, improvement and discharge, automatic discharge, and death. Data were statistically analyzed with independent sample or paired sample t test, Mann-Whitney U test, chi-square test, and Fisher's exact probability test. Results: During the 8 hours since fluid replacement, the actual input volume of electrolyte of all the children was significantly more than the planned input volume, and the actual input volumes of colloid, water, and total fluid were significantly less than the planned input volumes ( Z =13.094, 5.096, 13.256, 7.742, P <0.01). During the first and second 24 hours after injury, the actual input volumes of electrolyte of all the children were significantly more than the planned input volumes, and the actual input volumes of water and total fluid were significantly less than the planned input volumes ( Z =13.288, -13.252, 3.867, 13.183, -13.191, 10.091, P <0.01), while the actual input volumes of colloid were close to the planned input volumes ( P >0.05). During the 8 hours since fluid replacement, compared with those in unsatisfactory urine output maintenance group, there was no significant change in electrolyte coefficient or colloid coefficient of children in satisfactory urine output maintenance group ( P >0.05), while the water coefficient was significantly increased ( Z =2.574, P <0.05). Among children <3 years old, compared with those in >25%TBSA group, the electrolyte coefficient and water coefficient of children were significantly increased and the urine output of children was significantly decreased in 15%-25%TBSA group during the first and second 24 hours after injury ( Z =-3.867, -6.993, -3.417, -5.396, -5.062, 1.503, P <0.05 or P <0.01), while the colloid coefficient did not change significantly ( P >0.05); the levels of efficacy indicators of hematocrit, platelet count, and hemoglobin at 48 h after injury were significantly increased, while ALT level was significantly decreased ( Z =-2.720, -3.099, -2.063, -2.481, P <0.05 or P <0.01); the levels of the rest of the efficacy indicators and non-invasive monitoring indicators at 48 h after injury did not change significantly ( P >0.05). Among children aged 3-12 years, compared with those in >25%TBSA group, the electrolyte coefficient and water coefficient of children in 15%-25%TBSA group were significantly increased during the first and second 24 hours after injury, the colloid coefficient during the second 24 h was significantly decreased ( Z =-2.042, -4.884, -2.297, -3.448, -2.480, P <0.05 or P <0.01), while the colloid coefficient during the first 24 hours after injury, urine output during the first and second 24 hours after injury, and the non-invasive monitoring indicators and efficacy indicators at 48 hours after injury did not change significantly ( P >0.05). Complications occurred in 17 children during the treatment. Among the 235 children, 211 cases were cured, accounting for 89.79%, 5 cases were improved and discharged, accounting for 2.13%, 16 cases were discharged automatically, accounting for 6.81%, and 3 cases died, accounting for 1.28%. Conclusions: The electrolyte volume in early fluid resuscitation in severely burned children exceeding the volume calculated by the formula can obtain a good therapeutic effect. Among children <3 years old, the volume of fluid resuscitation should be appropriately increased in children with extremely severe burns compared with children with severe burns during fluid resuscitation; among children aged 3-12 years, the colloid volume should be appropriately increased in children with extremely severe burns compared with children with severe burns during fluid resuscitation; non-invasive monitoring indicators can be used to monitor hemodynamics and guide fluid resuscitation in severely burned children.

To evaluate the "flow void" diameter in patients with pregnancy-related diseases with and without uterine AVMs and assess the diagnostic performance of unenhanced MRI for uterine AVMs. From May 2014 to April 2019, 79 patients with pregnancy-related diseases were included, including 36 with and 43 without uterine AVMs confirmed by DSA. On MRI, the diameter of the most prominent "flow void" (hereinafter referred to as fv-D) was measured and compared between patients with and without uterine AVMs. The diagnostic performance of fv-D was estimated with receiver operating characteristic curves. The "flow void" sign was observed in patients with and without uterine AVMs (P > 0.05). The fv-D was significantly larger in patients with uterine AVMs in the myometrium and parametrium than in patients without uterine AVMs (P < 0.0001). The fv-D achieved a reliable diagnostic performance in the myometrium (sensitivity 80.6%, specificity 60.5%, negative predictive value 78.8%, positive predictive value 63%, AUC 0.727, cut-off: > 1.33 mm) and parametrium (sensitivity 97.2%, specificity 67.4%, negative predictive value 96.7%, positive predictive value 71.4%, AUC 0.881, cut-off > 2.6 mm). On MRI, fv-D could diagnose uterine AVMs. The fv-D had a much higher diagnostic efficiency in the parametrium than in the myometrium. The parametrium fv-D greatly improved the diagnostic sensitivity and provides a more accurate, noninvasive method of investigating possible uterine AVMs., (© 2021. The Author(s).)

Objectives: This study aims to explore the effect of acidic culture conditions on the proliferation, apoptosis, and migration ability of human tongue squamous cell carcinoma SCC15 and CAL27 cells and its potential molecular mechanism., Methods: After acidic culture for different periods, methyl thiazolyl tetrazolium (MTT) method was adop-ted to detect the cell proliferation of SCC15 and CAL27. Flow cytometry was employed to detect the apoptosis level of SCC15 and CAL27 cells. The migration ability of SCC15 and CAL27 after acidic culture was detected by scratch hea-ling test. Real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) was used to detect the mRNA expression of cyclooxygenase 2 (COX-2) and survivin in SCC15 and CAL27 cells after acidic culture., Results: After culture for 24 h under acidic microenvironment, SCC15 and CAL27 cells grew rapidly and reached the stationary phase after adjustment for 3 days. The apoptosis levels of SCC15 and CAL27 cells decreased after acidic culture, but the most significant reduction occurred after 6 h of acidic culture. The scratch healing rates of SCC15 and CAL27 cells increased after acidic culture. The results of FQ-PCR showed that the mRNA expression levels of COX-2 and survivin in SCC15 and CAL27 cells increased after acidic culture., Conclusions: Extracellular acidic microenvironment can inhibit the apoptosis of tongue squamous carcinoma cells, promote their migration, and induce more adaptable and malignant tongue squamous carcinoma cells. The mechanism may be related to COX-2 and survivin and their signal pathways.

Oral squamous cell carcinoma (OSCC) is one of the causes of cancer-related death worldwide. The abnormal proliferation ability of OSCC has become one of the major reasons for its poor prognosis. FK-506 binding protein 11 (FKBP11) is abnormally expressed in malignant tumors and affects many biological processes. The purpose of this study is to investigate the effect of FKBP11 on cell proliferation in OSCC and explore the possible regulatory mechanism. The expression of FKBP11 was detected by western blotting (WB) and/or real-time PCR in OSCC and paracancerous normal tissues in tongue squamous cell carcinoma (TSCC) cell lines, revealing high expression in OSCC and CAL-27 cells. Furthermore, FKBP11 knockdown inhibited the proliferation of CAL-27 cells by CCK-8 and colony formation assays. G 2 /M arrest and induction of apoptosis were observed using flow cytometry, Hoechst 33258 and Calcein-AM/PI staining, accompanied by changes in some cell cycle- and apoptosis-related proteins, including CDK1, Cyclin B1, p21, p27, p53, Bax, Bcl-2 and Caspase-3. Additionally, the expression of these proteins can be reversed by the use of pifithrin-α (PFT-α), a p53 inhibitor. An in vivo xenograft model further confirmed that FKBP11 enhanced OSCC progression. In conclusion, FKBP11 could promote cell proliferation by regulating G 2 /M phase and apoptosis via the p53/p21/p27 and p53/Bcl-2/Bax pathways, respectively, which suggests that it may be a new candidate target for the treatment of OSCC., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2021 Elsevier Inc. All rights reserved.)

GLUT4 is involved in rapid glucose uptake among various kinds of cells to contribute to glucose homeostasis. Prior data have reported that aberrant glucose metabolism by GLUT4 dysfunction in the uterus could be responsible for infertility and increased miscarriage. However, the expression and precise functions of GLUT4 in the endometrium under physiological conditions remain unknown or controversial. In this study, we observed that GLUT4 exhibits a spatiotemporal expression in mouse uterus on pregnant days 1-4; its expression especially increased on pregnant day 4 during the window of implantation. We also determined that estrogen, in conjunction with progesterone, promotes the expression of GLUT4 in the endometrial epithelium in vivo or in vitro . GLUT4 is an important transporter that mediates glucose transport in endometrial epithelial cells (EECs) in vitro or in vivo . In vitro , glucose uptake decreased in mouse EECs when the cells were treated with GLUT4 small interfering RNA (siRNA). In vivo , the injection of GLUT4-siRNA into one side of the mouse uterine horns resulted in an increased glucose concentration in the uterine fluid on pregnant day 4, although it was still lower than in blood, and impaired endometrial receptivity by inhibiting pinopode formation and the expressions of leukemia inhibitory factor (LIF) and integrin ανβ3, finally affecting embryonic development and implantation. Overall, the obtained results indicate that GLUT4 in the endometrial epithelium affects embryo development by altering glucose concentration in the uterine fluid. It can also affect implantation by impairing endometrial receptivity due to dysfunction of GLUT4., 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 © 2021 Long, Wang, Yuan, Dai, Liao, Zhang, Zhang, Ma, Lei, Cui, Zhang, Nie and Yue.)

Abstract: Objective To explore the feasibility of the CT image reconstruction of laryngeal cartilage and hyoid bone in adult age estimation using data mining methods. Methods The neck thin slice CT scans of 413 individuals aged 18 to <80 years were collected and divided into test set and train set, randomly. According to grading methods such as TURK et al., all samples were graded comprehensively. The process of thyroid cartilage ossification was divided into 6 stages, the process of cricoid cartilage ossification was divided into 5 stages, and the synosteosis between the greater horn of hyoid and hyoid body was divided into 3 stages. Multiple linear regression model, support vector regression model, and Bayesian ridge regression model were developed for adult age estimation by scikit-learn 0.17 machine learning kit （Python language）. Leave-one-out cross-validation and the test set were used to further evaluate performance of the models. Results All indicators were moderately or poorly associated with age. The model with the highest accuracy in male age estimation was the support vector regression model, with a mean absolute error of 8.67 years, much higher than the other two models. The model with the highest accuracy in female adult age estimation was the support vector regression model, with a mean absolute error of 12.69 years, but its accuracy differences with the other two models had no statistical significance. Conclusion Data mining technology can improve the accuracy of adult age estimation, but the accuracy of adult age estimation based on laryngeal cartilage and hyoid bone is still not satisfactory, so it should be combined with other indicators in practice., 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© by the Editorial Department of Journal of Forensic Medicine.)

Background: Metatarsalgia refers to localized or generalized forefoot pain in the region of the metatarsal heads. Often this pain is plantar, beneath the metatarsal heads, and arises from either mechanical or iatrogenic causes. The treatment of metatarsalgia remains controversial. A thorough understanding of the biomechanics of the forefoot and the underlying pathology of the particular type of metatarsalgia affecting the patient is a prerequisite to selecting the proper treatment. In recent years, massage therapy has been increasingly accepted by patients due to its lower costs, fewer unwanted side effects, and safety for clinical use. In this systematic review, we aim to evaluate the effectiveness and safety of massage therapy for patients with metatarsal pain., Methods: We will search the following electronic databases for randomized controlled trials to evaluate the effectiveness and safety of massage therapy in treating metatarsal pain: Wanfang and PubMed Database, CNKI, CENTRAL, CINAHL, and EMBASE. Each database will be searched from inception to October 2020. The entire process will include study selection, data extraction, risk of bias assessment, and meta-analyses., Results: This proposed study will evaluate the effectiveness and safety of massage therapy for patients with metatarsal pain. The outcomes will include changes in metatarsal pain relief and adverse effect., Conclusions: This proposed systematic review will evaluate the existing evidence on the effectiveness and safety of massage therapy for patients with metatarsalgia., Dissemination and Ethics: The results of this review will be disseminated through peer-reviewed publication. Because all of the data used in this systematic review and meta-analysis has been published, this review does not require ethical approval. Furthermore, all data will be analyzed anonymously during the review process., Osf Registration Number: DOI 10.17605/OSF.IO/C6KFJ.

Abstract: Objective To establish the orthogonal partial least square （OPLS） model for the estimation of early postmortem interval （PMI） of asphyxial death rats in four ambient temperatures based on gas chromatography-mass spectrometry （GC-MS） metabolomics. Methods The 96 rats were divided into four temperature groups （5 ℃, 15 ℃, 25 ℃ and 35 ℃）. Each temperature group was further divided into 3 h, 6 h, 12 h and 24 h after death, and 6 other rats were taken as the control group. The cardiac blood was collected at the set time points for the four temperature groups and 0 h after death for the control group for the metabolomics analysis by GC-MS. By OPLS analysis, the variable importance in projection （VIP）>1 and the result of Kruskal-Wallis test P<0.001 were used to screen out the differential metabolite related to PMIs in the cardiac blood of rats of different temperature groups. Then OPLS regression models of different temperature groups were established with these metabolites. At the same time, a prediction group for investigating the prediction ability of these models was set up. Results Through the analysis of OPLS, 18, 15, 24 and 30 differential metabolites （including organic acids, amino acids, sugars and lipids） were screened out from the rats in groups of 5 ℃, 15 ℃, 25 ℃ and 35 ℃, respectively. The prediction results of the four temperature group models showed that the prediction deviation of 5 ℃ model was larger than that of other groups. The prediction results of other temperature groups were satisfactory. Conclusion There are some differences in the changes of metabolites in cardiac blood of rats at different ambient temperatures. The influence of ambient temperature should be investigated in the study of PMI estimation by metabolomics, which may improve the accuracy of PMI estimation., 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© by the Editorial Department of Journal of Forensic Medicine.)

Background: Chronic atrophic gastritis (CAG) is an established precursor of gastric carcinoma with high prevalence worldwide. It is a typical complex gastro-intestinal disease with multiple influence factors, of which exact mechanisms remain unelucidated. Therefore, an ideal strategy to relieve CAG is urgently needed. In recent years, massage therapy has been increasingly accepted by CAG patients due to its lower costs, fewer unwanted side effects and safety for clinical use. In this systematic review, we aim to evaluate the effectiveness and safety of massage therapy for patients with chronic atrophic gastritis., Methods: We will search the following electronic databases for randomized controlled trials to evaluate the effectiveness and safety of massage therapy in treating chronic atrophic gastritis: Wanfang and Pubmed Database, China National Knowledge Infrastructure Database, Cochrane Central register of controlled trials, Cumulative Index of Nursing and Allied Health Literature, and Excerpta Medica database. Each database will be searched from inception to September 2020. The entire process will include study selection, data extraction, risk of bias assessment, and meta-analyses., Result: This proposed study will evaluate the effectiveness and safety of massage therapy for patients with chronic atrophic gastritis. The outcomes will include changes in CAG relief and adverse effect., Conclusion: This proposed systematic review will evaluate the existing evidence on the effectiveness and safety of massage therapy for patients with chronic atrophic gastritis., Dissemination and Ethics: The results of this review will be disseminated through peer-reviewed publication. Because all of the data used in this systematic review and meta-analysis has been published, this review does not require ethical approval. Furthermore, all data will be analyzed anonymously during the review process.

Abstract: Adult age determination plays an important role in individual identification, criminal investigation and social welfare. The most popular adult age determination indicators are pubic symphysis, iliac auricular surface, costal cartilage, cranial sutures, teeth, laryngeal cartilage, etc. In recent years, with the progress of CT imaging and 3D reconstruction technology, the adult age determination study gradually has transferred from a time-consuming general observation of bones with complex pre-processing in the past to the non-destructive, convenient, time-saving and easy to store image analysis technology. To explore more accurate, rapid and convenient adult age determination methods, multiple imaging methods and artificial intelligence have been applied in adult age determination. This paper reviews the common methods and research progress of adult age determination at home and abroad, infers the development direction of adult age determination, in order to provide reference for the improvement and optimization of forensic adult age determination., 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© by the Editorial Department of Journal of Forensic Medicine.)

Acupuncture is widely used in the treatment of cerebral hemorrhage, and it improves outcomes in experimental animal models and patients. However, the mechanisms underlying the effectiveness of acupuncture treatment for cerebral hemorrhage are still unclear. In this study, a model of intracerebral hemorrhage was produced by injecting 50 μL autologous blood into the caudate nucleus in Wistar rats. Acupuncture at Baihui (DU20) and Qubin (GB7) acupoints was performed at a depth of 1.0 inch, 12 hours after blood injection, once every 24 hours. The needle was rotated at 200 r/min for 5 minutes, For each 30-minute session, needling at 200 r/min was performed for three sessions, each lasting 5 minutes. For the positive control group, at 6 hours, and 1, 2, 3 and 7 days after induction of hemorrhage, the rats were intraperitoneally injected with 1 mL aniracetam (0.75 mg/mL), three times a day. The Bederson behavioral test was used to assess palsy in the contralateral limbs. Western blot assay was used to examine the expression levels of Nestin and basic fibroblast growth factor in the basal ganglia. Immunohistochemistry was performed to count the number of Nestin- and glial cell line-derived neurotrophic factor-positive cells in the basal ganglia. Acupuncture effectively reduced hemorrhage and brain edema, elevated the expression levels of Nestin and basic fibroblast growth factor in the basal ganglia, and increased the number of Nestin- and glial cell line-derived neurotrophic factor-positive cells in the basal ganglia. Together, these findings suggest that acupuncture promotes functional recovery after cerebral hemorrhage by increasing the expression of neurotrophic factors. The study was approved by the Committee for Experimental Animals of Heilongjiang Medical Laboratory Animal Center (approval No. 2017061001) on June 10, 2017., Competing Interests: None

The anaerobic conversion efficiency of excess sludge in China is significantly lower than that in developed countries. Sludge characteristics are important influencing factors in the anaerobic degradability of sludge. The typical differences between excess sludge in China and in developed countries are mainly reflected in three aspects:long sludge age, high content of micro-sized grits, and high content of metal ions. Therefore, the effects of sludge age, micro-sized grits (represented by silica particles), and metal ions on the anaerobic digestion (AD) of excess sludge were studied systematically. Quantified comparison and targeted enhancement research were also carried out. The results showed that the negative effect of grit in the influent on VS reduction of sludge during AD was not obvious, while sludge age and metal ions were the main inhibitory factors. In addition, the inhibition level of sludge age was significantly higher than that of metal ions ( P <0.05). With an increase in sludge age, the inhibition level of metal ions showed a further strengthening trend. Therefore, it could be concluded that long sludge age was the most important factor limiting the degradability of excess sludge. Moreover, with thermal hydrolysis pretreatment (160℃, 0.55 MPa, 30 min), the cumulative methane yield and VS reduction of sludge with long sludge age (40 d) almost increased to the values of sludge with a sludge age of 5 d. That is, thermal hydrolysis pretreatment has the potential to break through the bottleneck of degradability of sludge caused by long sludge age.

Objective: MicroRNAs (miRNAs) are important regulators in the progression of lung adenocarcinoma (LAD). Moreover, microRNA-149 (miR-149) exhibits different roles in human cancers. Hence, this study mainly focused on the function of miR-149 in LAD., Patients and Methods: Western blot analysis and Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) were used to quantify expression levels. The regulatory mechanism of miR-149/RAP1B was explored by methyl thiazolyl tetrazolium (MTT), transwell, and Dual-Luciferase reporter assays., Results: Downregulation of miR-149 was detected in LAD and predicted worse prognosis in patients with LAD. Functionally, overexpression of miR-149 inhibited cell viability and metastasis in LAD. In addition, miR-149 directly targets RAP1B and restrained its expression in LAD. Furthermore, upregulation of RAP1B attenuated the inhibitory effect of miR-149 on LAD. Besides that, miR-149 blocked epithelial-mesenchymal transition (EMT) and Wnt/β-catenin pathway in LAD., Conclusions: MiR-149 inhibited the progression of LAD by restraining RAP1B/EMT and inactivating Wnt/β-catenin pathway.

The third generation of the CRISPR/Cas9-mediated genome fixed-point editing technology has been widely used in the field of gene editing and gene expression regulation. How to improve the on-target efficiency and specificity of this system, as well as reduce its off-target effects are always the bottleneck in its development. Machine learning provides novel methods to the problems of the CRISPR/Cas9 system, and CRISPR/Cas9-based machine learning has recently become a very hot research topic. In this review, we firstly outline the mechanism of the CRISPR/Cas9 system. Subsequently, we elaborate the current issues of CRISPR/Cas9, including low efficiency and potential off-target effects, and sequence-recognizing limitation from protospacer adjacent motif (PAM). Finally, we summarize the applications of methods within the machine learning framework for optimizing the CRISPR/Cas9 system, such as optimized single-guide RNA (sgRNA) design, CRISPR/Cas9 cleavage efficiency prediction, off-target effects evaluation, gene knock-out as well as high-throughput functional genetic screening and prospects for development.

Baihui (DU20)-penetrating-Qubin (GB7) acupuncture can inhibit inflammatory reactions and activate signaling pathways related to proliferation after intracerebral hemorrhage. However, there is no research showing the relationship between this treatment and cell apoptosis. Rat models of intracerebral hemorrhage were established by injecting 60 μL of autologous blood into the right side of the caudate-putamen. Six hours later, the needle traveled subcutaneously from the Baihui acupoint to Qubin acupoint. The needle was alternately rotated (180 ± 10 turns/min) manually along clockwise and counter-clockwise directions. Stimulation lasted for 7 days, and was performed three times each for 6 minutes with 6-minute intervals between stimulations. Rats intraperitoneally receiving Sonic hedgehog pathway activator, purmorphamine (1 mg/kg per day), served as positive controls. Motor and sensory function were assessed using the Ludmila Belayev test. Extent of pathological changes were measured in the perihemorrhagic penumbra using hematoxylin-eosin staining. Apoptosis was examined by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling assay. Expression of smoothened (Smo) and glioma-associated homolog 1 (Gli1) was determined by western blot assay. Our results showed that Baihui-penetrating-Qubin acupuncture promoted recovery of motor and sensory function, reduced the apoptotic cell percentage in the perihemorrhagic penumbra, and up-regulated Smo and Gli1 expression. We conclude that Baihui-penetrating-Qubin acupuncture can mitigate hemorrhage and promote functional recovery of the brain in a rat model of intracerebral hemorrhage, possibly by activating the Sonic hedgehog pathway., Competing Interests: None declared

Inflammation plays an important role in nerve defects caused by intracerebral hemorrhage. Repairing brain damage by inhibiting the macrophage-inducible C-type lectin/spleen tyrosine kinase (Mincle/Syk) signaling pathway is a potential new target for treating cerebral hemorrhage. In this study, we aimed to determine whether acupuncture through Baihui (DU20) to Qubin (GB7) is an effective treatment for intracerebral hemorrhage through the Mincle/Syk signaling pathway. An intracerebral hemorrhage rat model was established by autologous blood infusion into the caudate nucleus. Acupuncture through Baihui to Qubin was performed for 30 minutes, once every 12 hours, for a total of three times. Piceatannol (34.62 mg/kg), a Syk inhibitor, was intraperitoneally injected as a control. Modified neurological severity score was used to assess neurological function. Brain water content was measured. Immunohistochemistry and western blot assay were used to detect immunoreactivity and protein expression levels of Mincle, Syk, and CARD9. Real-time polymerase chain reaction was used to determine interleukin-1β mRNA levels. Hematoxylin-eosin staining was performed to observe histopathological changes. Our results showed that acupuncture through Baihui to Qubin remarkably improved neurological function and brain water content, and inhibited immunoreactivity and expression of Mincle, Syk, CARD9, and interkeukin-1β. Moreover, this effect was similar to piceatannol. These findings suggest that acupuncture through Baihui to Qubin can improve neurological impairment after cerebral hemorrhage by inhibiting the Mincle/Syk signaling pathway., Competing Interests: None declared

Digested sludge, as the main by-product of the sewage sludge anaerobic digestion process, still contains considerable organic compounds. In this protocol, we report a facile method for preparing digested sludge-derived self-doped porous carbon material for high-performance supercapacitor electrodes via a sustainable pyrolysis/activation process. The obtained digested sludge-derived carbon material (HPDSC) exhibits versatile O-, N-doped hierarchical porous framework, high specific surface area (2103.6 m 2  g -1 ) and partial graphitization phase, which can facilitate ion transport, provide more storage sites for electrolyte ions and enhance the conductivity of active electrode materials. The HPDSC-based supercapacitor electrodes show favourable energy storage performance, with a specific capacitance of 245 F g -1 at 1.0 A g -1 in 0.5 M Na 2 SO 4 ; outstanding cycling stability, with 98.4% capacitance retention after 2000 cycles; and good rate performance (211 F g -1 at 11 A g -1 ). This work provides a unique self-doped three-dimensional hierarchical porous carbon material with a favourable charge storage capacity and at the same time finds a high value-added and environment-friendly strategy for disposal and recycling of digested sludge., Competing Interests: We declare we have no competing interests.

To improve anaerobic digestion biogas yield, the removal of organic-bonding metals by EDTA pretreatment was investigated to strengthen the anaerobic biogas effect. The results showed that after EDTA pretreatment (experimental group), the organic-bonding metals were reduced from (5.09±0.57)% to (1.37±0.20)% (calculated with TS) and dissolved organic matter increased significantly (SCOD improved 627%), suggesting that this method could remove a large amount of organic-bonding metals and enhance the dissolution of organic sludge. According to the results for the apparent activation energy (AAE) of sludge organic matter dissolution, the AAE of the experiment group was 36% lower than the control group, showing that pretreatment could effectively reduce the reaction energy barrier for organic matter dissolution. In a test of anaerobic acid production lasting 16 days, the VFAs of the experiment group were higher than the control group, with the largest upgrade of 42%. In the anaerobic methane production test (BMP) that lasted for 22 days, compared with control group, the accumulated biogas production of the experiment group increased by 48%. Through further research on the dynamics of methane production, the rate-limiting step in the experiment group was methanogenesis, while that of the control group was hydrolysis, indicating that removal of organic-bonding metals by EDTA pretreatment could effectively accelerate the hydrolysis rate.

Objective: To explore nicotinamide phosphoribosyltransferase (NAMPT) expression in dilated cardiomyopathy (DCM) and its initial mechanism in the pathogenesis of DCM., Methods: The peripheral blood of 131 Chinese patients with DCM confirmed by West China Hospital of Sichuan University during 2010-2013 were collected. 137 cases of Chinese Han healthy persons who were randomly selected in the physical examination center of West China Hospital of Sichuan University as the control group. The serum NAMPT levels were measured by ELISA. The NAMPT mRNA levels were determined by RT-PCR. Plasmids over-expressing NAMPT and empty vector were constructed and transfected into H9C2 cells. By using WST-1 technique,cell cycle detection and flow cytometry measurements,the effect of NAMPT on H9C2 proliferation and apoptosis was studied., Results: Serum NAMPT level was significantly higher in the DCM group compared with that of controls and positively associated with the grade of heart failure and the size of left ventricular in DCM patients. The NAMPT mRNA level was significantly lower in the DCM group than that in the control group. The plasmid over-expressing NAMPT promoted H9C2 cells proliferation and increased the proportion of S phase cells compared with that of empty plasmid group. Over-expressing NAMPT increased proportion of the viable cells and reduced the proportion of late apoptotic and necrotic cells than empty plasmid group in the basic situation or after being treated with different concentrations of H 2 O 2 ., Conclusion: The high expression of plasma protein level of NAMPT while low expression of NAMPT mRNA in peripheral blood cells,contributes one of the biological characteristics to DCM. The decrease of intracellular NAMPT may be an important factor in the pathogenesis of DCM., (CopyrightCopyright© by Editorial Board of Journal of Sichuan University (Medical Science Edition).)

Two nanostructured poly(sulfosalicylic acid) (PSA) films were synthesized from room temperature ionic liquid (RTIL) or aqueous solution on a glassy carbon electrode (GCE) via potentiodynamic electropolymerization. The morphology and properties of the PSA films were characterized with scanning electron microscopy (SEM), scanning probe microscopy (SPM), electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). It was found that solvent had a major influence on the morphology and electrochemical properties of the resultant PSA films. The PSA(Ι) film, which was prepared from RTIL, consists of granular particles with cracks, whereas the PSA(II) film prepared from aqueous solution consists of nano-triangles with a more compact surface. The blocking effect of the PSA(Ι) film for the [Fe(CN) 6 ] 3-/4- electrochemical probe is much stronger, and a remarkably enhanced voltammetric response of the [Ru(NH 3 ) 6 ] 3+ electrochemical probe can be observed for the PSA(II) film. When it is used to detect dopamine in the presence of a high concentration of ascorbic acid, PSA(II)/GCE has three linear parts with better discrimination and a detection limit of 0.03μM. For PSA(Ι)/GCE, there are two linear parts with a detection limit of 0.05μM. However, the reproducibility and storage stability of PSA(Ι)/GCE are better than those of PSA(ΙI)/GCE., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Biochemical methane potential experiments were conducted to investigate the effects of organic loading rate on the performance of anaerobic digesters with vinegar residues. According to the comparisons of methane production and liquid phase compositions, as well as thermogravimetry, X-ray Diffraction and infrared spectroscopy analyses, the conclusions could be drawn that:①Lower organic loading rate was better to mitigate the accumulation of VFAs and pH decrease during the hydrolysis and acidification of organic matters, which consequently improved methane production. When the inoculum to substrate ratio was 1:1[organic loading rate of 1.78 g·(L·d) -1 , pH=7.60], the cumulative methane production was the highest, reaching 2249.7 mL, and the performance of the digesters was stable. The VFAs content increased with the increase of the organic loading rate, leading to the suppression and further the stop of methane production. And when the inoculum to substrate ratio was 1:4[organic loading rate of 7.12 g·(L·d) -1 , pH=5.52], the simultaneous generation of acetate and lactic acids could be achieved at 8000 mg·L -1 and 2650 mg·L -1 , respectively. ② As vinegar residues were short-range ordered with microcrystalline structure or mainly contained amorphous substances, they were more biodegradable than feedstocks such as corn stalk. During the anaerobic digestion processes, the degradation rates of lignin, cellulose and hemicellulose increased with the decrease of organic loading rate.

High free ammonia nitrogen (FAN) is regarded as a main factor that inhibits biogas production in high solid anaerobic digestion systems with dewatered sludge. Two mesophilic semi-continuous anaerobic digesters fed with dewatered sludge (input total solids=15%) were operated and their performance under different FAN concentrations by two regulation methods including pH reduction and total ammonia nitrogen (TAN) promotion were investigated. Results showed that when FAN was elevated from (400±174) mg·L -1 to (526±25) mg·L -1 with the increase of TAN, the daily biogas yield declined from (11.9±0.3) L·d -1 to (10.3±0.2) L·d -1 . Meanwhile, volatile solids (VS) reduction rate decreased by 33.7%. When FAN dropped from (330±99) mg·L -1 to (47±13) mg·L -1 due to pH reduction, daily biogas yield declined from (14.4±1.1) L·d -1 to (10.8±0.3) L·d -1 and VS reduction rate decreased by 26.9%. The system performance was not enhanced with the reduction of FAN. With the stop of pH regulation, FAN gradually raised again to 300 mg·L -1 and digester performance recovered to the same as the situation before regulation. The result suggested that inhibition occurred during pH reduction. By 16S rRNA pyrosequencing analysis on bacterial and archaeal diversity and quantitative PCR analysis aiming at methanogens, it was found that bacterial community structure both changed obviously with obvious performance decline in the two digesters. Excessive FAN (>500 mg·L -1 ) could change the main bacteria species which degrade protein from Tepidimicrobium and Proteiniborus to Anaerobranca . On the other hand, some carbohydrate and protein degrading bacteria were also inhibited by pH reduction for FAN control, leading to the decline of hydrolysis rates of organic matters. As a result, the shortage of substrate supply for the methanogens weakened the biogas production of the digestion system.

Objective: To investigate the clinical effects of traditional Chinese medicine (TCM) fumigation on type ⅢA prostatitis with the symptoms of damp heat and blood stasis., Methods: We randomly divided 72 cases of type ⅢA prostatitis with the symptoms of damp heat and blood stasis into an experimental and a control group of equal number, the former treated with TCM fumigation plus oral Levofloxacin while the latter with oral Levofloxacin only. After 4 weeks of treatment, we compared the National Institute of Health Chronic Prostatitis Symptom Index (NIHCPSI) score, TCM symptoms score, maxinum flow rate (Qmax), and average urinary flow rate (Qave) between the two groups of patients., Results: The total effectiveness rate was significantly higher in the experimental than in the control group (91.7% vs 61.1%, P< 0.01), while the NIHCPSI score in the experimental group was markedly decreased after treatment as compared with the baseline (14.5 ± 8.2 vs 26.5 ± 9.3, P< 0.05) and significantly lower than that in the control (14.5 ± 8.2 vs 20.6 ± 7.9, P< 0.05). Qmax was remarkably increased in the experimental group after treatment in comparison with the baseline (［21.2 ± 4.3］ vs ［15.8 ± 3.6］ ml/s, P< 0.05) and the control group, and so was Qave as compared with pretreatment (［16.3 ± 3.5］ vs ［10.5 ± 2.8］ ml/s, P< 0.05) and the control. However, the patients of the control group showed no statistically significant differences before and after treatment either in Qmax (［15.4 ± 3.4］ vs ［16.1 ± 2.9］ ml/s, P>0.05) or in Qave (［10.9 ± 2.4］ vs ［11.1 ± 2.9］ ml/s, P>0.05)., Conclusions: TCM fumigation combined with Levofloxacin is an effective therapy for type ⅢA prostatitis with the symptoms of damp heat and blood stasis, which can significantly improve the symptoms, reduce the NIHCPSI score, and increase the Qmax and Qave of the patient.

Objective: To observe the effect of penetrative needling of "Baihui" (GV 20) to "Qubin" (GB 7) on neurologic functions and expression of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and toll-like receptor 4 (TLR-4, involving in inflammatory reactions) in the tissue around the local cerebral hematoma in rats with intracerebral hemorrhage (ICH), so as to provide evidence for clinical treatment of ICH., Methods: Fifty-four male Wistar rats were randomly divided into sham control, mo-del and acupuncture groups, and then further divided into three time-point subgroups(1,3,7 days after modeling, n =6/subgroup). The ICH model was established by injection of the rat's autoblood (50 μL) into the putaman region (P:0.2 mm, R:3.5 mm) in a stereotaxic apparatus and confirmed by Berderson's neurologic examination grading system (0-3 points). The neurologic function was assessed by using Longa's scoring (5-points) and footfault asymmetry testing[footfault index=(contra faults-ipsi faults)/total steps in 2 min]. For penetrative needling, an acupuncture needle was inserted into GV 20 and controlled to advance to GB 7 on the affected side and retained for 30 min, once daily. The expression of TNF-α, IL-6 and TLR-4 in the cerebral tissue around the putaman was detected by immunohistochemistry., Results: After penetrative needling stimulation, the increased Longa's score and footfault asymmetry score in ICH rats were significantly decreased on day 1, 3 and 7 after modeling ( P <0.01), suggesting an improvement of neurologic function after the treatment. Immunohistochemical staining outcomes of the cerebral tissue surrounding the autoblood injection site showed that the expression levels of TNF-α, IL-6 and TLR-4 proteins on day 1, 3 and 7 were considerably higher in the model group than in the control group ( P <0.01), and markedly lower in the acupuncture group than in the model group ( P <0.01), suggesting a suppression of the proinflammatory factors and TLR-4 levels around the locus of the brain after needling intervention. A positive correlation existed between the expression levels of TLR-4 and IL-6/TNF-α., Conclusions: Penetrative needling stimulation of GV 20 to GB 7 can reduce the levels of proinflammatory factors TNF-α and IL-6, and TLR-4 in the ICH tissues in rats with cerebral hemorrhage, which may contribute to its effect in improving neurological function.

Background: Type 2 diabetes mellitus, obesity and hepatic steatosis showed a strong correlation with metabolic syndrome. However, data on the influence of pancreatic steatosis on metabolic syndrome are lacking., Objective: Our aim is to perform the prevalence of pancreatic steatosis in adults and its association with metabolic syndrome in a Chinese population., Methods: This was a cross-sectional study, randomly selected. A total of 1190 health examination subjects were recruited. Pancreatic steatosis or hepatic steatosis was diagnosed via trans-abdominal sonography. The clinical and metabolic parameters were compared between the two groups, and their associations with pancreatic steatosis were examined., Results: The prevalence of pancreatic steatosis was 30.7%. The presence of pancreatic steatosis was significantly increased by age, gender, central obesity, hepatic steatosis, hypertriglyceridemia and hyperglycemia. In the logistic regression analysis, age (P < 0.05), central obesity (P < 0.01), diabetes (P < 0.05), hypertriglyceridemia (P < 0.05) and hepatic steatosis (P < 0.01) were independently associated with pancreatic steatosis. The number of the parameters of the metabolic syndrome in pancreatic steatosis group was more than that in non-pancreatic steatosis group [(2.5 ± 1.1) vs (1.4 ± 1.2)] (P < 0.01)., Conclusion: The pancreatic steatosis is strongly associated with the parameters of metabolic syndrome, such as central obesity, diabetes, and hepatic steatosis., (Copyright © 2016 IAP and EPC. Published by Elsevier B.V. All rights reserved.)

Developing efficient, low-cost, and stable carbon-based catalysts for oxygen reduction reaction (ORR) to replace the expensive platinum-based electrocatalysts remains a major challenge that hamper the practical application of fuel cells. Here, we report that N, Fe, and S co-doped nanoporous carbon material, derived via a facile one-step pyrolysis of sewage sludge, the major byproduct of wastewater treatment, can serve as an effective electrocatalyst for ORR. Except for the comparable catalytic activity with commercial 20% Pt/C via a nearly four-electron transfer pathway in both alkaline and acid medium, the as-synthesized co-doped electrocatalyst also exhibits excellent methanol crossover resistance and outstanding long-term operation stability. The organic compounds in sewage sludge act as the carbon source and the in-situ N and S dopant in the fabrication, while the inorganic compounds serve as the in-built template and the in-situ Fe dopant. Our protocol demonstrates a new approach in the economic and eco-friendly benign reuse of sewage sludge, and also provides a straightforward route for synthesizing excellent carbon-based electrocatalysts as promising candidates for ORR directly from a type of waste/pollution.

Fluoroquinolone antibiotics (FQs), as the common pharmaceuticals and personal care products (PPCPs), are widespread in the environment. FQs contained in wastewater would be ultimately enriched in sludge, posing a potential threat to the consequent sludge utilization. To optimize the analytical method applicable to the determination of FQs in sludge, the authors selected ofloxacin (OFL), norfioxacin (NOR), ciprofloxacin (CIP) and lomefloxacin (LOM) as the target FQs, and established a method which was based on cell lysis, FQs extraction with triethylamine/methanol/water solution, Solid Phase Extraction (SPE) and HPLC-Fluorescence Detection (FLD) determination. After the investigation, phosphoric acid-triethylamine was decided to be the buffer salt, and methanol was chosen as the organic mobile phase. The gradient fluorescence scanning strategy was proved to be necessary for the optimal detection as well. Furthermore, by the designed orthogonal experiments, the effects of the extraction materials, pH, and the eluents on the efficiency of SPE extraction were evaluated, by which the optimal extraction conditions were determined. As a result, FQs in liquid samples could be analyzed by utilizing HLB extraction cartridge, and the recovery rates of the four FQs were in the range of 82%-103%. As for solid samples, the recovery rates of the four FQs contained reached up to 71%-101%. Finally, the adsorptivity of the sludge from the different tanks ( anaerobic, anoxic and oxic tanks) was investigated, showing gradual decrease in the adsorption capacity, but all adsorbed over 90% of the EQs. This conclusion also confirmed that 50% removal of FQs in the domestic wastewater treatment plant was realized by sludge adsorption.

Objective: To explore the value of estimating chronologic age based on the grades of mandibular third molar development. To evaluate whether mandibular third molar could be used as an indicator for estimating the age under or over 18 years., Methods: The mineralization status of mandibular third molar of 1 845 individuals aged 10 - 30 was graded and marked based on Demirjian's classification of grades reformed by Orhan. Gender difference was examined by t-test. A cubic regression model was established to analyze the correlation between third molar and chronologic age. Each grade of age cumulative distribution diagram and ROC curve was respectively performed to evaluate the relationship between third molar and the age of 18. Using Bayes discriminant analysis, an equation was established for estimating the age of 18., Results: The inner-rater reliability was 0.903. Statistical analysis showed a moderate correlation between age and grade. Significant differences of both genders were found only in grade D and H (P < 0.05). Males at the grades from 1 to D and females at the grades from 1 to C were under 18 years old, and both males and females at grade H were over 18 years old. The area under the ROC curve was 0.797 (P < 0.05)., Conclusion: Third molar development shows a high correlation with age, and combined with other indicators, it can be used to estimate the age of 18.

We investigated gene expression in embryonic stem (ES) cells, induced pluripotent stem (iPS) cells, and fibroblasts. Microarray expression data sets obtained from the Gene Expression Omnibus were analyzed using the Partek software. Human genes from ES cells, iPS cells, and fibroblasts were ranked from low to high according to their expression levels. The gene expression mode in iPS cells was much more like the mode in ES cells, and the expression levels of fibroblast genes fluctuated more dramatically than those of iPS and ES cells. The number of genes with significantly different expression was lower in the iPS and ES cells. Several genes with the expression levels that were significantly different between ES and iPS cells were found, including LEFTY2, DLK1, and NLRP2. Four genes belonged to the low expression category in fibroblasts with the high expression category occurring in ES cells, i.e., HESRG, PROM1, NTS, and LRRN1. Analyzing the expression of these genes is helpful to elucidate the mechanisms of cell fate regulation and efficiently obtain iPS cells.

Identification of epigenetic alterations in tumors has become a common method for identifying genes critical to cancer development and progression. Thus, we identified DNA methylation alterations on the genome scale during lung adenocarcinoma (LADC) progression to understand the carcinogenic process and identify clinically relevant biomarkers. We found that epigenetic alterations in LADC mainly occur during the early stage of LADC progression, and there are no significant methylation differences between early-stage and late-stage LADCs. This suggests that DNA methylation alterations characterize a turning point of early events in LADC progression. By comparing DNA methylation between early-stage LADCs and normal lung tissues, we further identified 940 genes with significant alterations in DNA methylation. Sixty-seven genes were found to exhibit strong correlation between methylation alterations and expression changes, based on associated gene expression data. According to gene ontology analysis, these genes are involved in lung development, respiratory system development, cell cycle, histidine metabolism, the Wnt signaling pathway, and the p53 signaling pathway. We also found that genes on chromosome 18 most frequently showed promoter hypermethylation. Moreover, we found that LADC-associated DNA hypomethylation occurred preferentially at neither histone H3 lysine 4 nor histone H3 lysine 27 mark domains in human embryonic stem cells (NMDs) and that hypomethylation of NMDs was associated with a poor prognostic signature in LADC. Our findings have important implications for LADC progression because of the identification of novel epigenetic biomarkers potentially involved in early-stage LADC and for establishing the importance of NMD DNA hypomethylation for predicting prognosis in LADC.

Notch pathway activation maintains neural stem cells in a proliferating state and increases nerve repair capacity. To date, studies have rarely focused on changes or damage to signal transduction pathways during cerebral hemorrhage. Here, we examined the effect of acupuncture in a rat model of cerebral hemorrhage. We examined four groups: in the control group, rats received no treatment. In the model group, cerebral hemorrhage models were established by infusing non-heparinized blood into the brain. In the acupuncture group, modeled rats had Baihui (DU20) and Qubin (GB7) acupoints treated once a day for 30 minutes. In the DAPT group, modeled rats had 0.15 μg/mL DAPT solution (10 mL) infused into the brain. Immunohistochemistry and western blot results showed that acupuncture effectively inhibits Notch1 and Hes1 protein expression in rat basal ganglia. These inhibitory effects were identical to DAPT, a Notch signaling pathway inhibitor. Our results suggest that acupuncture has a neuroprotective effect on cerebral hemorrhage by inhibiting Notch-Hes signaling pathway transduction in rat basal ganglia after cerebral hemorrhage.

Objective: Nucleosomes are the basic packaging units of chromatin, determinants of nucleosome organization playing a major role in genome packaging. Although a wide variety of nucleosome organization factors have been considered separately across the whole or partial human genomic regions, it is unclarified that what the major determinants and their roles in scale are when being put all together. And it is also unknown that what the similarities and differences of determinants between different genomic features such as genes of different expression levels or genomic regions with different functions., Materials and Methods: We detected commonalities and characteristics of nucleosome positioning determinants in different genes and regions with 1591486 nucleosomes identified by ourselves in human CD4+ cell., Results: It was found that a distinct linear combination of about 20 nucleosome-positioning factors explained nucleosome occupancy for each genomic feature. In those linear combinations, 6 DNA sequence attributes (Roll stiffness and Twist stiffness, CT and AG, CG and shift stiffness) and a histone modification (H4R3me2) are shared. And other factors are varied. Roll stiffness and Twist stiffness are the most important features. They are dominant, alone explaining 96.61-98.45% of the positioning weight in each genomic feature. The characteristic factors in each combination are larger in number, but weaker in power. Numerous histone modifications play a subtle role for nucleosome positioning., Conclusions: The present study provides a more accurate positioning nucleosome-map with higher resolution and a dramatically simplified means to predict and understand intrinsic nucleosome occupancy in different genomic features in human CD4+ cell. Roll stiffness and Twist stiffness are the two most important determinants in all genomic features. They may dominate because they both determine the degree of DNA bending and correlates with many other DNA structural characteristics. Histone modifications play a role of subtle allocation for nucleosome occupancy.

In this research, a new method for sewage sludge (SS) disposal was introduced, by which heavy metals were implanted into sewage sludge before pyrolysis. Cu was adopted as the representative of heavy metals to test this process and was implanted in the form of CuCl2. Effects of Cu implanting concentration and reaction temperature on the residual ratio and immobilization of heavy metals in pyrolysis char were studied. Meanwhile, two leaching methods were employed with the purpose to determine the maximum capacity of heavy metal immobilization in the char. The primary research results showed that when the Cu implanting concentration was 0.5% (mass fraction), more than 90% of Cu remained in the char after carbonization, and the leachability of heavy metals in the char was related to pyrolysis temperature. Cu leaching from the char increased with increasing pyrolysis temperature. There was also a limitation for Cu implanting concentration in the sewage sludge, which was determined by the destination of the pyrolyzed char. If it went to sanitary landfill, the limitation would be 0.5%. The primary results showed that sewage sludge could be kneaded with other wastes containing heavy metals before pyrolysis to achieve co-processing.