[Advances in the study of the actin nucleation factor Spire].

There are three main classes of actin nucleation factors: Arp2/3 complexes, Spire and Formin. Spire assembles microfilaments by nucleating stable longitudinal tetramers and binding actin to the growing end of the microfilament. As early as 1999, Wellington et al. identified Spire as an actin nucleating agent, however, over the years, most studies have focused on Arp2/3 and Formin proteins; there has been relatively less research on Spire as a member of the actin nucleating factors. Recent studies have shown that Spire is involved in the vesicular transport through the synthesis of actin and plays an important role in neural development. In this paper, we reviewed the structure, expression and function of Spire, and its association with disease in order to identify meaningful potential directions for studies on Spire.

Multi-Omics Research Accelerates the Clarification of the Formation Mechanism and the Influence of Leaf Color Variation in Tea (Camellia sinensis) Plants.

Tea is a popular beverage with characteristic functional and flavor qualities, known to be rich in bioactive metabolites such as tea polyphenols and theanine. Recently, tea varieties with variations in leaf color have been widely used in agriculture production due to their potential advantages in terms of tea quality. Numerous studies have used genome, transcriptome, metabolome, proteome, and lipidome methods to uncover the causes of leaf color variations and investigate their impacts on the accumulation of crucial bioactive metabolites in tea plants. Through a comprehensive review of various omics investigations, we note that decreased expression levels of critical genes in the biosynthesis of chlorophyll and carotenoids, activated chlorophyll degradation, and an impaired photosynthetic chain function are related to the chlorina phenotype in tea plants. For purple-leaf tea, increased expression levels of late biosynthetic genes in the flavonoid synthesis pathway and anthocyanin transport genes are the major and common causes of purple coloration. We have also summarized the influence of leaf color variation on amino acid, polyphenol, and lipid contents and put forward possible causes of these metabolic changes. Finally, this review further proposes the research demands in this field in the future.

Percutaneous Intra-arterial Hyaluronidase Injection for Hyaluronic Acid Filler Embolism Threatening Skin Barrier Integrity: Implementation of a Stepwise Treatment Protocol.

BACKGROUND: Hyaluronic acid (HA) filler-induced vascular embolism that threatens skin integrity is an urgent situation. There is increasing evidence that percutaneous intra-arterial hyaluronidase injection is an effective therapeutic technique for it. However, until now, there is a lack of a unifying protocol about the technique. OBJECTIVES: This study aims to provide a conclusion of percutaneous intra-arterial hyaluronidase injection along with adjunctive measures on the treatment of occlusions precipitated by HA-based filler and develop a stepwise treatment protocol. METHODS: We searched PubMed for peer-reviewed studies, consensus statements, case series, and case reports using a variety of keywords. RESULTS: High-dose, pulsed hyaluronidase is the mainstay for the treatment of HA filler-induced embolism, but percutaneous intra-arterial hyaluronidase injection is a more effective technique. Until now, hyaluronidase is injected into three arteries percutaneously, including facial artery, supratrochlear artery, and superficial temporal artery. Furthermore, the adjunctive measures that may optimize clearance of an occlusion and/or skin barrier repair such as the use of image guidance and CGF should be considered. CONCLUSION: Vascular occlusions that threaten skin integrity are an urgent matter which requires accurate diagnosis and effective intervention. Percutaneous intra-arterial hyaluronidase injection along with adjunctive measures performed in a stepwise manner is key to an optimal outcome. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

7-Methoxy-13-dehydroxypaxilline: New indole diterpenoid from an endophytic fungus Penicillium sp. Nb 19.

ABSTACTA chemical investigation of the endophyte Penicillium sp. Nb 19, isolated from leaves of the traditionally medical plant Baphicacanthus cusia (Nees) Bremek., yielded one new indole diterpenoid, 7-methoxy-13-dehydroxypaxilline (1) together with seven known metabolites (2-8). The obtained structure of compound 1 was elucidated by its spectroscopic data. In addition, the absolute configuration of compound 6 was confirmed by ECD for the first time. Compounds 1-6 were evaluated for antitumor activity against MCF-7, HepG2, and HCCC-9810 cell lines.

Perioperative care of nipple-areola complex-sparing mastectomy and one-stage breast reconstruction via endoscopic axillary approach for ductal carcinoma in situ: A case report.

RATIONALE: Breast cancer represents a prevalent malignancy that primarily impacts women, with pronounced consequences on their overarching health. The major therapeutic approach, encompassing surgical procedures, can often culminate in mastectomy, potentially inciting psychological turmoil and disorders. PATIENT CONCERNS: A patient was admitted to our facility on May 5, 2023, precipitated by the discovery of bilateral breast masses during a routine physical examination conducted 3 days before admission. DIAGNOSIS: The breasts were symmetric, with the right nipple inverted and a palpable mass in the upper outer quadrant of the right breast, measuring approximately 5 cm × 4 cm. The mass was firm with indistinct borders, relatively regular morphology, poor mobility, and no tenderness. Outpatient color Doppler ultrasound revealed heterogeneous echogenicity in the right breast, classified as Breast Imaging Reporting and Data System (BI-RADS) category 0, along with multiple ductal dilatations. The left breast exhibited a hypoechoic area (BI-RADS 3), indicative of proliferative changes. Radiographic mammography confirmed diffuse changes in the right breast (BI-RADS 0) and proliferative signs in the left breast (BI-RADS 2). Biopsy results reveal significant atypical ductal hyperplasia consistent with intermediate-grade ductal carcinoma in situ. This patient was diagnosed as ductal carcinoma in situ of the right breast (cTisN0M0 and Stage 0), accompanied by a left breast mass. INTERVENTIONS: On May 15, 2023, the patient was readmitted for further surgical intervention. Following relevant auxiliary examinations, the patient underwent nipple-areola complex-sparing radical mastectomy for the right breast, sentinel lymph node biopsy in the right axillary area, prosthesis-based breast reconstruction for the right breast, and microrotatotomy of the left breast mass on the left side on May 17. OUTCOMES: The patient made a successful recovery under scrupulous perioperative supervision and was discharged 7 days post-surgery. LESSONS: The axillary approach for endoscopic mammary gland excision and immediate implant reconstruction permits patients to preserve the esthetics of the female form while undergoing conventional medical treatment. This methodology considerably enhances the psychophysical health of the patients, thereby marking it as an advantageous practice worthy of broad dissemination in the medical community.

Factors affecting the accumulation of organotins by wild fish: A case study in the Three Gorges Reservoir, China.

Organotin compounds (OTs) accumulate in fish easily, however, research on their influencing factors is still limited. This study collected 25 species of fish with different diets, habitats, and age from the Three Gorges Reservoir (TGR), the largest deep-water river channel-type reservoir in China, and analyzed the accumulation characteristics of OTs in these fish. The results showed that tributyltin (TBT) and triphenyltin (TPhT) were the dominant OTs in fish from the TGR. The correlation between OTs concentration and age, body length, and body weight varied with fish species. The concentrations of TBT and TPhT in carnivorous fish (mean, 25.78 and 11.69 ng Sn/g dw, respectively) were higher than those in other diet fish (P<0.01), but there was no significant difference in fish at different habitat water layers (P>0.05). In addition, the degradation rates of TBT and TPhT in different fish species were all below 50%. In summary, the accumulation of TBT and TPhT in fish is mainly influenced by diet, and both TBT and TPhT were difficult to degrade in fish. These results reveal the pollution characteristics of OTs in fish from the TGR, and can improve our understanding of the factors influencing TBT and TPhT accumulation in freshwater fish.

M4IDP stimulates ROS elevation through inhibition of mevalonate pathway and pentose phosphate pathway to inhibit colon cancer cells.

Maintaining redox homeostasis is an essential feature of cancer cells, and disrupting this homeostasis to cause oxidative stress and induce cell death is an important strategy in cancer therapy. M4IDP, a zoledronic acid derivative, can cause the death of human colorectal cancer cells by increasing the level of intracellular reactive oxygen species (ROS). However, its potential molecular mechanism is unclear. Our in vitro studies showed that treatment with M4IDP promoted oxidative stress in HCT116 cells, as measured by the decreased ratios of GSH/GSSG and NADPH/NADP+ and increased level of MDA. M4IDP could cause the decrease of GSH content, the increase of GSSG content, the decrease of NADPH content and pentose phosphate pathway flux, the downregulation of G6PD expression, the upregulation of unprenylated Rap1A and total expression of RhoA and CDC42. The increase of ROS and cytotoxicity induced by M4IDP could be reversed by the supplementation of NADPH, the overexpression of G6PD and the supplementation of GGOH. In vivo studies showed that M4IDP inhibited tumor growth in the human colorectal cancer xenograft mouse model, which was accompanied with a decreased [18F]FDG uptake. Collectively, these results provide evidence that M4IDP can promote oxidation in colon cancer cells by inhibiting mevalonate pathway and pentose phosphate pathway and produce therapeutic effect. This study revealed for the first time a possible mechanism of bisphosphonate-induced increase of ROS in malignant tumor cells. This is helpful for the development of new molecular therapeutic targets and can provide new ideas for the combined therapy of bisphosphonates in tumors.

Modulation of gut microbiota alleviates cerebral ischemia/reperfusion injury in rats by inhibiting M1 polarization of microglia.

Gut microbiota affects the gut-brain axis; hence, the modulation of the microbiota has been proposed as a potential therapeutic strategy for cerebral ischemia/reperfusion injury (CIRI). However, the role and mechanism of the gut microbiota in regulating microglial polarization during CIRI remain poorly understood. Herein, using a middle cerebral artery occlusion and reperfusion (MCAO/R) rat model, we evaluated changes in the gut microbiota after CIRI and the potential effects of fecal microbiota transplant (FMT) on the brain. Rats underwent either MCAO/R or sham surgery, and then they received FMT (started 3 days later; continued for 10 days). 2,3,5-Triphenyltetrazolium chloride staining, neurological outcome scale, and Fluoro-Jade C staining showed that MCAO/R induced cerebral infarction, neurological deficits, and neuronal degeneration. In addition, immunohistochemistry or real-time PCR assay showed increased expression levels of M1-macrophage markers-TNF-α, IL-1ß, IL-6, and iNOS-in the rats following MCAO/R. Our finding suggests that microglial M1 polarization is involved in CIRI. 16 S ribosomal RNA gene sequencing data revealed an imbalance in the gut microbiota of MCAO/R animals. In contrast, FMT reversed this MCAO/R-induced imbalance in the gut microbiota and ameliorated nerve injury. In addition, FMT prevented the upregulation in the ERK and NF-κB pathways, which reversed the M2-to-M1 microglial shift 10 days after MCAO/R injury in rats. Our primary data showed that the modulation of the gut microbiota can attenuate CIRI in rats by inhibiting microglial M1 polarization through the ERK and NF-κB pathways. However, an understanding of the underlying mechanism requires further study.

Study on Virulence Genes, Drug Resistance and Molecular Epidemiology of Klebsiella pneumoniae with High Virulence in Inner Mongolia, China.

Objective: The purpose of this study was to analyse the clinical, microbiological and molecular epidemiological characteristics of patients with pyogenic liver abscess (PLA) caused by Klebsiella pneumoniae (KPN) in Inner Mongolia, China. Methods: The KPN isolates from 78 KPN-PLA cases admitted to a tertiary teaching hospital in Baotou, Inner Mongolia, from 2016 to 2019 were studied systematically and described comprehensively. The virulence factors, drug resistance and sequence types of KPN in different samples were identified by a wire-drawing test, polymerase chain reaction, a drug susceptibility test and multi-site sequence typing. Results: There were more male than female KPN-PLA patients (P<0.05). The mortality rate was 2.5%, and KPN-PLA was significantly associated with diabetes mellitus (P<0.05). Most of the KPN isolates in the puncture fluid of patients with KPN-PLA were hypervirulent KPN (HvKP). The positive rate of the KPN-PLA specimens was higher than that of the blood and urine specimens. The KPN isolates of the urine specimens had higher drug resistance than the other two (P<0.05). The hypermucoviscous KPN, aerobic actin (aero) (+), K1 and K2 serotypes accounted for 80.8%, 89.7%, 56.4% and 26.9%, respectively. In addition to ironB (3.8%), the detection rates of virulence factors rmpA, irp2, entB, iucD, aero, wcaG, iutA, kfu, ybtA, iron, fimH and mrkD were higher (69.2%-100.0%). The positive rate of KPN isolates of the KPN-PLA puncture fluid was higher than that of the blood and urine samples (P<0.05). In addition, ST23 was found to be the dominant ST (32.1%) of KPN-PLA in the Baotou region. Conclusion: In the KPN-PLA specimens, the KPN isolates were more virulent than those in the blood and urine specimens, and a carbapenem-resistant HvKP strain emerged. This research will help improve the understanding of HvKP and provide useful suggestions for KPN-PLA treatments.

Granzyme B: A novel therapeutic target for treatment of atopic dermatitis.

Granzyme B is a serine protease that can play multiple roles in intracellular and extracellular perforin-dependent or non-perforin-dependent mechanisms. Granzyme B has been found to be an important factor involved in the pathogenesis of atopic dermatitis and is increased in both skin lesions and peripheral blood of atopic dermatitis patients. In this article, we review the correlation between granzyme B and atopic dermatitis to provide a novel therapeutic targeting option for clinical treatment of the latter.

Non-coding RNAs: Role in diabetic foot and wound healing.

Diabetic foot ulcer (DFU) and poor wound healing are chronic complications in patients with diabetes. The increasing incidence of DFU has resulted in huge pressure worldwide. Diagnosing and treating this condition are therefore of great importance to control morbidity and improve prognosis. Finding new markers with potential diagnostic and therapeutic utility in DFU has gathered increasing interest. Wound healing is a process divided into three stages: Inflammation, proliferation, and regeneration. Non-coding RNAs (ncRNAs), which are small protected molecules transcribed from the genome without protein translation function, have emerged as important regulators of diabetes complications. The deregulation of ncRNAs may be linked to accelerated DFU development and delayed wound healing. Moreover, ncRNAs can be used for therapeutic purposes in diabetic wound healing. Herein, we summarize the role of microRNAs, long ncRNAs, and circular RNAs in diverse stages of DFU wound healing and their potential use as novel therapeutic targets.

Delayed diagnosis and management of necrotizing fasciitis of the left lower leg: A case report.

INTRODUCTION: Necrotizing fasciitis (NF) is a rare, severe soft tissue infection, characterized by rapid and extensive necrosis of the skin, subcutaneous tissue, and superficial and deep fascia. It is frequently misdiagnosed as other infectious diseases, leading to inappropriate treatment and potentially serious consequences. It may be complicated by septic shock and multiple organ failure with a fatal outcome. PATIENT CONCERNS: A 73-year-old woman presented with continuous itching, skin lesions, pain, and swelling of the outer side of her left leg. The patient was diagnosed with septic shock and multiorgan failure caused by left leg NF. DIAGNOSIS: Septic shock and multiorgan failure caused by left leg NF. INTERVENTIONS: Two surgeries were performed on the patient's leg, which effectively treated her septic shock and multiple organ dysfunction. OUTCOMES: The patient was followed up three times after her discharge. She had a good recovery, was generally well with no significant sequelae, and returned to her regular life. CONCLUSION: NF is an acute severe illness with high mortality. It is easily misdiagnosed, leading to delayed or erroneous treatment and serious (or potentially fatal) outcomes. Rapid and accurate diagnosis of NF is essential for patient recovery. In difficult cases, multidisciplinary consultations may be helpful. The management of NF includes early and thorough surgical debridement, antibiotics, and symptomatic treatment.

Successful management of a breastfeeding mother with severe eczema of the nipple beginning from puberty: A case report.

BACKGROUND: Nipple eczema is the most common presentation of atopic dermatitis of the breast, which seriously influences breastfeeding of mothers. We here present a case of severe nipple eczema that started in puberty and received continuous care and interventions during pregnancy. The patient succeeded in breastfeeding after the interventions. CASE SUMMARY: A 36-year-old woman at 16 wk of gestation (gestation 1 parturition 0), visited the breastfeeding consultation clinic, complaining of excessive nipple secretion, severe itching, and concerns about breastfeeding. She was diagnosed with severe nipple eczema. Health education, consultation with dermatologists, topical medication, psychological support, and postpartum care were carried out. Through continuous interventions, her nipple eczema significantly improved, lactating confidence enhanced, anxiety symptoms were reduced, and exclusive breastfeeding was achieved. CONCLUSION: For lactating women with nipple eczema, breastfeeding consultants should play an important role in patient education and provide whole-process and individual guidance.

Effect of ammonia-oxidizing bacterial strain that survives drought stress on corn compensatory growth upon post-drought rewatering.

A pot experiment was performed under rain-shelter conditions to explore the effects of drought stress and post-drought rewatering on the abundance of an ammonia-oxidizing bacteria (AOB) strain in corn (Zea mays L.) rhizosphere soils and the relationship between the AOB strain and corn (Zea mays L.) compensatory growth after drought stress rewatering. Corn seedlings were used as test materials, and one AOB strain was isolated and screened from the soil. The experimental design included six treatments: (1) wet (WT), (2) wet with AOB strain inoculation during wetness (WI), (3) wet with AOB strain inoculation during rewatering (WR), (4) post-drought rewatering (DT), (5) post-drought rewatering with AOB strain inoculation during wetness (DI), and (6) post-drought rewatering with AOB strain inoculation during rewatering (DR). Wetness and drought stress were obtained by keeping the soil water content at 75-80% and 50-55% of the field capacities, respectively. The results showed that the isolated and screened AOB strain (S2_8_1) had 100% similarity to Ensifer sesbaniae. The inoculation of S2_8_1 during the wet period in the DI treatment caused it to colonize the rhizosphere soil. Drought stress decreased its abundance, but rewatering resulted in a great increase. The S2_8_1 in the DI treatment increased the total biomass, water use efficiencies, net photosynthetic rates, rhizosphere soil nitrification rates, leaf cytokinin concentrations, xylem sap cytokinin concentrations, copy number of S2_8_1 in rhizosphere soils, and organic carbon contents in rhizosphere soils by 23, 104, 35, 30, 18, 29, 104, and 23% on day 10 after rewatering compared with WT treatment. In the DI treatment, the increase in rhizosphere soil nitrification rates caused by S2_8_1 during wetness was closely related to the cytokinin delivery from roots to leaves and increased leaf cytokinin concentrations. The increase in leaf cytokinin concentrations improved rewatering corn growth, which caused compensatory growth and increased water use. Compensatory and over-compensatory growths occurred in DT and DR treatments, respectively. Therefore, the coexistence of the strain of AOB with corn in rhizosphere soil increased the corn compensatory growth by regulating soil nitrification and root-induced leaf cytokinin.

Oxidative Damage to BV2 Cells by Trichloroacetic Acid: Protective Role of Boron via the p53 Pathway.

This study aimed to investigate the neurotoxicity induced by trichloroacetic acid (TCA) and the possible protective mechanisms of boron (B). Mouse BV2 cells were treated with TCA (0, 0.39, 0.78, 1.56, 3.12, 6.25, or 12.5 mmol/L) and B (0, 7.8, 15.6, 31.25, 62.5, 125, 500, or 1,000 mmol/L) for 3 h and 24 h, respectively. Then, reactive oxygen species, and supernatant proinflammatory cytokine and protein levels were analyzed after 24 h of combined exposure. Beyond the dose-dependent decrease in the cellular viability, it clearly increased after B supplementation ( P < 0.05). Moreover, B decreased oxidative damage, and significantly down-regulated IL-6 levels and up-regulated TNF-ß production ( P < 0.05). B also decreased apoptosis via the p53 pathway. The present findings indicated that TCA may induce oxidative damage, whereas B mitigates these adverse effects by decreasing cell apoptosis.

Organotins in a food web from the Three Gorges Reservoir, China: Trophic enrichment and potential health risk.

Triphenyltin (TPhT) and tributyltin (TBT) remain widely present in various aquatic environments despite restrictions on their use in many countries for many years. The biomagnification of these compounds in the aquatic food web remains controversial. This study reports the bioaccumulation of TPhT and TBT in aquatic animals in the Three Gorges Reservoir (TGR), a deep-water river channel-type reservoir and the largest reservoir in China. We measured TPhT, TBT and their metabolites in 2 invertebrates, 27 fish and the aquatic environment. The logarithmic bioaccumulation factors of TPhT and TBT were 4.37 and 3.77, respectively, indicating that TPhT and TBT were enriched in organisms of the TGR. Both TPhT and TBT concentrations were significantly and positively correlated with trophic level, with trophic magnification factors of 3.71 and 3.63, respectively, indicating that TPhT and TBT exhibited similar trophic enrichment in the freshwater food web of the TGR. The results of health risk assessment showed that although all hazard index (HI) values were <1, more attention should be paid to the health risk to children associated with consumption of aquatic products (HI = 0.67). This study provides powerful evidence of trophic enrichment of TPhT and TBT in a freshwater food web in a deep-water river channel-type reservoir and provides valuable data regarding organotins in aquatic animals in the TGR.

Occurrence, seasonal variation, potential sources, and risks of organophosphate esters in a cold rural area in Northeast China.

Organophosphate esters (OPEs) in the environment have been the focus of increasing attention due to their ubiquity and potential toxicity. However, there is little information on the occurrence and characteristics of OPEs in rural areas, especially those with cold year-round temperatures and frozen soil in winter. In this study, environmental samples were collected, in summer and winter, from villages and towns in Northeast China differing in the types and intensities of their anthropogenic activities. The samples were analyzed for 12 OPEs. The results showed the widespread presence of alkyl-OPEs, Cl-OPEs, and aryl-OPEs in the water, soil, snow, and ice of the study sites. In summer, tris(1-chloro-2-propyl) phosphate (TCPP) and tris(2-chloroethyl) phosphate (TCEP) were the primary compounds in water and soil, respectively. The ∑12OPE concentration in three villages varied from 46.26 to 257.37 ng/L in water, and from 6.62 to 19.46 ng/g in soils. The ∑12OPE concentrations in water were lower in winter than summer, but conversely, ∑12OPE concentrations in frozen soils in winter were higher than those in soils in summer. In winter, there was a shift in the predominant OPEs in water and frozen soils, with dominance of TCEP and complex compounds, respectively. Obvious seasonal characteristics of the potential sources and ecological risks of OPEs in these areas were also determined, with more complex sources of OPEs seen in summer than winter. In summer, only 2-ethylhexyl diphenyl phosphate (EHDPP) in water posed a potential risk, while in summer and, especially, in winter, EHDPP and tris(2-ethylhexyl) phosphate posed potential risks in soils. The high ∑12OPE concentration in snow (56.77 ng/L) implied that wet deposition can amplify OPEs in other environmental compartments. This is the first systematic report on OPEs in a cold rural area. Our findings highlight the need for seasonal monitoring of OPEs in similar areas.

Efficacy of Percutaneous Intraarterial Facial/Supratrochlear Arterial Hyaluronidase Injection for Treatment of Vascular Embolism Resulting From Hyaluronic Acid Filler Cosmetic Injection.

BACKGROUND: Vascular embolism is a serious complication of hyaluronic acid (HA) filler cosmetic injection, and hyaluronidase injection has been proposed as the treatment. Until now, there has been a lack of adequate clinical evidence regarding the benefits of treatment for HA filler-induced vascular embolism by percutaneous facial or supratrochlear arterial hyaluronidase injection. OBJECTIVES: The authors sough to evaluate the efficacy of percutaneous facial or supratrochlear arterial hyaluronidase injection as a rescue treatment for HA filler-induced vascular embolism. METHODS: We included 17 patients with vascular embolism after facial HA filler injection. Intraarterial injection of 1500 units hyaluronidase was performed via facial artery for 13 cases with skin necrosis and via supratrochlear arterial for 4 cases with severe ptosis and skin necrosis but no visual impairment. Simultaneously, general symptomatic treatment and nutritional therapy were performed. RESULTS: After hyaluronidase injection, facial skin necrosis in all cases was restored and ptosis in the 4 cases was also significantly relieved. Patients were subsequently followed-up for 1 month to 1 year. The skin necrosis in 16 patients completely healed, and only 1 patient had small superficial scars. CONCLUSIONS: It is effective to alleviate skin necrosis and ptosis resulting from HA filler embolism via percutaneous facial or supratrochlear arterial hyaluronidase injection.

Accounting for the cost of ecological degradation in Fuzhou City, China.

Ecological degradation accounting is a critical content of building green GDP and gross economic-ecological product (GEEP) accounting systems. With ecosystems in Fuzhou City as a research object, we built an accounting framework for the cost of ecological degradation according to the unreasonable human activity. Following the accounting framework, we calculated the ecological degradation cost in Fuzhou City of 2015 and 2018. The results showed that the ecological degradation cost in Fuzhou City of 2015 was 9.08 billion yuan, accounting for 1.6% of local GDP. For different ecosystem types, marine ecological degradation cost was the largest, about 6.311 billion yuan, accounting for 69.5% of the total loss, followed by wetland ecosystem, accounting for 28.7%, and then arable land, contributing only 1.8%. In term of functions, the cost of provisioning degradation was the largest, about 6.313 billion yuan, accounting for 69.5% of the total loss, followed by regulation loss, accounting for 27.5%, mainly from the loss of climate regulation and hydrological regulation. In the regional distribution, the ecological degradation cost was mainly concentrated in Lianjiang County, Luoyuan County, Fuqing City, and Minqing County, up to 8.092 billion yuan in total, accounting for 89.7% of total loss. Compared with 2015, the ecological degradation cost in 2018 decreased by 2.608 billion yuan, showing an obvious downward trend, with a decrease rate of 28.7%. The reductions were major in Lianjiang County (86.4%), Luoyuan County (14.8%), Fuqing City (19.9%), and Minqing County (12.6%), and mainly concentrated in marine and wetland ecosystems. Such a result indicated that people's awareness of ecological protection in Fuzhou City was increasing, and that the ecological damage due to human activity in marine and wetland system was obviously decreased. This study provided data support for promoting regional sustainable development and ecological civilization construction.

Developing Two Rapid Protein Extraction Methods Using Focused-Ultrasonication and Zirconia-Silica Beads for Filamentous Fungi Identification by MALDI-TOF MS.

Filamentous fungi identification by Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been challenging due to the lack of simple and rapid protein extraction methods and insufficient species coverage in the database. In this study, we created two rapid protein extraction methods for filamentous fungi: a one-step zirconia-silica beads method (ZSB) and a focused-ultrasonication method (FUS). The identification accuracy of two methods were evaluated with the VITEK MS, as well as number of spectra peaks and signal-to-noise ratio (S/N) with M-Discover 100 MALDI-TOF MS compared to the routine method. The better method was applied to build a filamentous fungi in-house spectra library for the M-Discover 100 MS, and then another one and routine method were performed in parallel to verify the accuracy and commonality of the in-house library. Using the two optimized methods, the dedicated operating time before MALDI-TOF MS analysis was reduced from 30 min to 7 (ZSB) or 5 (FUS) min per sample, with only a few seconds added for each additional strain. And both two methods identified isolates from most mold types equal to or better than the routine method, and the total correct identification rate using VITEK MS was 79.67, 76.42, and 76.42%, respectively. On the other hand, the two rapid methods generally achieved higher maximum and minimum S/N ratios with these isolates tested as compared to the routine method. Besides, the ZSB method produced overall mean of maximum and minimum S/N ratio higher than that by FUS. An in-house library of M-Discover MS was successfully built from 135 isolates from 42 species belonging to 18 genera using the ZSB method. Analysis of 467 isolates resulted in 97.22% correctly identified isolates to the species level by the ZSB method versus 95.50% by the routine method. The two novel methods are time- and cost-effective and allow efficient identification of filamentous fungi while providing a simplified procedure to build an in-house library. Thus, more clinical laboratories may consider adopting MALDI-TOF MS for filamentous fungi identification in the future.