Homologous polydopamine ameliorates haemolysis of melittin for enhancing its anticancer efficacy.

Despite exhibiting potent anticancer activity, the strong hemolytic properties of melittin (MEL) significantly restrict its delivery efficiency and clinical applications. To address this issue, we have devised a strategy wherein homologous dopamine (DA), an essential component of bee venom, is harnessed as a vehicle for the synthesis of MEL-polydopamine (PDA) nanoparticles (MP NPs). The ingenious approach lies in the fact that MEL is a basic polypeptide, and the polymerization of DA is also conducted under alkaline conditions, indicating the distinctive advantages of PDA in MEL encapsulation. Furthermore, MP NPs are modified with folic acid to fabricate tumor-targeted nanomedicine (MPF NPs). MPF NPs can ameliorate the hemolysis of MEL in drug delivery and undergo degradation triggered by high levels of reactive oxygen species (ROS) within solid tumors, thereby facilitating MEL release and subsequent restoration of anticancer activity. After cellular uptake, MPF NPs induce cell apoptosis through the PI3K/Akt-mediated p53 signaling pathway. The tumor growth inhibitory rate of MPF NPs in FA receptor-positive 4T1 and CT26 xenograft mice reached 78.04% and 81.66%, which was significantly higher compared to that in FA receptor-negative HepG2 xenograft mice (45.79%). Homologous vehicles provide a new perspective for nanomedicine design.

Causal Relationship Between Endometriosis and Pelvic Inflammatory Diseases: Mendelian Randomization Study.

Objective: This study explores the causal relationship between endometriosis and pelvic inflammatory diseases (PID). Methods: The study utilized genome-wide association study (GWAS) datasets for endometriosis ("finn-b-N14_ENDOMETRIOSIS") and PID ("finn-b-N14_OTHFEMPELINF"). Subsequently, two-sample Mendelian randomization (MR) analyses were conducted using inverse variance weighting (IVW), Egger regression (MR-Egger), and weighted median (WM) methods. Heterogeneity was evaluated using Cochran's Q test, and in case of detected outliers, they were removed for re-evaluation of MR causality. Results: From the endometriosis GWAS dataset, 33 single nucleotide polymorphisms (SNPs) were selected as instrumental variables. All three methods, IVW (OR = 1.39, P < 1×10-8), MR-Egger (OR = 1.41, P = 0.003), and WM (OR = 1.37, P = 1.16×10-5) confirmed a causal relationship between endometriosis and PID. The association between endometriosis and pelvic inflammation remained unaffected by the exclusion of individual SNPs. Lastly, Cochran's Q test and funnel plots showed no evidence of SNP asymmetry. Conclusion: The results of the MR analysis support a potential causal relationship between endometriosis and an increased risk of PID.

Enhanced Piezocatalytic Performance of Li-doped BaTiO3 Through a Facile Sonication-Assisted Precipitation Approach.

Piezocatalysis-induced dye degradation has garnered significant attention as an effective method for addressing wastewater treatment challenges. In our study, we employed a room-temperature sonochemical method to synthesize piezoelectric barium titanate nanoparticles (BaTiO3: BTO) with varying levels of Li doping. This approach not only streamlined the sample preparation process but also significantly reduced the overall time required for synthesis, making it a highly efficient and practical method. One of the key findings was the exceptional performance of the Li-doped BTO nanoparticles. With 20 mg of Li additive, we achieved 90 % removal of Rhodamine B (RhB) dye within a relatively short timeframe of 150 minutes, all while subjecting the sample to ultrasonic vibration. This rapid and efficient dye degradation was further evidenced by the calculated kinetic rate constant, which indicated seven times faster degradation rate compared to pure BTO. The enhanced piezoelectric performance observed in the Li-doped BTO nanoparticles can be attributed to the strategic substitution of Li atoms, which facilitated a more efficient transfer of charge charges at the interface. Overall, our study underscores the potential of piezocatalysis coupled with advanced materials like Li-doped BTO nanoparticles as a viable and promising solution for wastewater treatment, offering both efficiency and environmental sustainability.

Correlation Analysis between Intrarenal Small Artery Intimal Thickening and Clinicopathological Features and Prognosis in Primary Membranous Nephropathy Patients.

BACKGROUND: Primary membranous nephropathy (PMN) is the most common pathological type of nephrotic syndrome in adults. Intrarenal small artery intimal thickening can be observed in most renal biopsies. The purpose of this study was to investigate the association between intrarenal small artery intimal thickening and clinicopathological features and prognosis in PMN patients. METHODS: Data were continuously collected from patients who were diagnosed with PMN in Shenzhen Second People's Hospital (The First Affiliated Hospital of Shenzhen University) from 2008 to 2021 for a retrospective cohort study. Regression analysis and survival analysis were used to analyze the relationship between intrarenal small artery intimal thickening and renal prognosis in PMN patients. RESULTS: 300 PMN patients were enrolled in this study, including 165 patients (55%) with intrarenal small artery intimal thickening. Patients with intimal thickening were older, with higher BMI, systolic blood pressure and diastolic blood pressure, serum uric acid, a higher proportion of hypertension, acute kidney injury, nephrotic syndrome, more urine protein, and lower eGFR. Multivariate Cox regression analysis showed that after adjusting for age, gender, hypertension, BMI, urine protein, eGFR, and the use of ACEI/ARB and hormone immunosuppressants, intimal thickening was a risk factor for renal prognosis in PMN patients (HR = 3.68, 95% CI 1.36-9.96, p < 0.05). Kaplan-Meier survival curve analysis showed that the incidence of reaching the renal composite outcome was higher in the intimal thickening group (p < 0.05). CONCLUSION: The prognosis of PMN patients with intrarenal small artery intimal thickening is worse, so early intervention is very important for these patients.

Importance of controlling mesocotyl elongation in the development of rice seedlings intended for mechanical transplantation.

The mesocotyl facilities the emergence of deep-sown rice. However, the effects of mesocotyl elongation on mechanically transplanted rice seedlings remain unclear. In this study, the indica three-line hybrid rice Chuanyou 6709 (CY6709) and the indica conventional rice Guichao II (GCII) were selected as experimental materials. The seedlings were grouped based on mesocotyl lengths of 1.0 and 2.0 cm (M1 and M2, respectively), and seedlings without mesocotyl elongation were used as a control (M0). Seedling morphology, root morphology and physiology, and dynamic changes in soluble sugar and protein, malondialdehyde (MDA), and antioxidant enzyme activity in the mesocotyl were evaluated. The results showed that the elongation of mesocotyl is not conducive to improving the quality of mechanically transplanted seedlings, resulting in weak seedlings and poor root coiling force. The mesocotyl lengths of the seedlings showed a single peak with increasing seedling age, which gradually disappeared. The longer the mesocotyls, the slower their senescence. The MDA content of M2 was significantly lower than that of M1, and the activities of soluble sugar, soluble protein, and antioxidant enzymes of M2 were higher than those of M1, implying that seedlings with longer mesocotyls yielded lower-quality seedlings, reducing their suitability for mechanized transplantation. Compared with those of M0, the root-shoot ratio, stem base width, leaf age, leaf area, white root number, root coiling force, root length, root surface area, and root volume of M1 and M2 were reduced. Therefore, in the raising of rice seedlings, excessive elongation of the rice mesocotyl is not conducive to optimum root growth and development of aboveground structures for seedlings that are suitable for mechanized transplantation. Controlling the mesocotyl elongation can facilitate the cultivation of high-quality mechanically transplanted seedlings.

Combination of Biochar and Trichoderma harzianum Can Improve the Phytoremediation Efficiency of Brassica juncea and the Rhizosphere Micro-Ecology in Cadmium and Arsenic Contaminated Soil.

Phytoremediation is an environment-friendly method for toxic elements remediation. The aim of this study was to improve the phytoremediation efficiency of Brassica juncea and the rhizosphere soil micro-ecology in cadmium (Cd) and arsenic (As) contaminated soil. A field experiment was conducted with six treatments, including a control treatment (CK), two treatments with two contents of Trichoderma harzianum (T1: 4.5 g m-2; T2: 9 g m-2), one biochar treatment (B: 750 g m-2), and two combined treatments of T1B and T2B. The results showed Trichoderma harzianum promoted the total chlorophyll and translocation factor of Brassica juncea, while biochar promoted plant biomass compared to CK. T2B treatment showed the best results, which significantly increased Cd accumulation by 187.49-308.92%, and As accumulation by 125.74-221.43%. As a result, the soil's total Cd content was reduced by 19.04% to 49.64% and total As contents by 38.76% to 53.77%. The combined amendment increased the contents of soil available potassium, phosphorus, nitrogen, and organic matter. Meanwhile, both the activity of glutathione and peroxidase enzymes in plants, together with urease and sucrase enzymes in soil, were increased. Firmicutes (dominant bacterial phylum) and Ascomycota (dominant fungal phylum) showed positive and close correlation with soil nutrients and plant potentially toxic elements contents. This study demonstrated that phytoremediation assisted by biochar and Trichoderma harzianum is an effective method of soil remediation and provides a new strategy for enhancing plant remediation efficiency.

Biancaea decapetala (Roth) O.Deg. extract exerts an anti-inflammatory effect by regulating the TNF/Akt/NF-κB pathway.

BACKGROUND: Biancaea decapetala (Roth) O.Deg. (Fabaceae) is used to treat colds, fever, and rheumatic pain caused by inflammation. However, the mechanism underlying its anti-inflammatory properties remains unclear. PURPOSE: This study aimed to evaluate the anti-inflammatory activity of Biancaea decapetala extract (BDE) in vitro and in vivo and explore the possible underlying mechanism and potential targets. METHODS: The release of nitric oxide (NO) and inflammatory cytokines in LPS-stimulated RAW264.7 cells and rats were measured using Griess reagent and enzyme-linked immunosorbent assay (ELISA). Hematoxylin and eosin (H&E) staining was employed to examine the pathology of animal tissues. Transcriptome analysis was performed to screen the pathways related to BDE-mediated inhibition of inflammation, and the expression of related proteins was measured using real-time quantitative polymerase chain reaction (RT-qPCR), western blotting, ELISA, and immunofluorescence methods. Surface Plasmon Resonance (SPR) and the Drug Affinity Reaction Target Stability (DARTS) method were used to verify whether BDE binds to TNF-α target protein, while a L929 cell model and NF-κB gene reporter systematic method were used to investigate the inhibitory effect of BDE on the activity of TNF-α protein. RESULTS: BDE inhibited the expression of TNF-α, IL-1ß, IL-6, and NO in RAW264.7 cells and rats, and improved the pathological changes in lung tissue. RNA-seq showed that BDE may regulate the TNF/Akt/NF-κB pathway to inhibit inflammation onset. BDE significantly downregulated the mRNA expression of TNF-α, IL-6, IL-1ß, and that of relevant proteins, including TNF-α, p-p65, p-Akt, p-IκBα. Furthermore, BDE inhibited the nuclear translocation of NF-κB (p65) and the activation of the Akt pathway by SC79. The L929 cell model, luciferase reporter gene analysis, DARTS, and SPR experiments showed that BDE may bind to TNF-α and inhibit the TNF-α-NF-κB pathway. CONCLUSION: BDE may target TNF-α to inhibit the TNF/Akt/NF-κB pathway, thereby attenuating inflammation. These findings reveal the anti-inflammatory effects and mechanisms of BDE and provide a theoretical basis for the further development and utilization of BDE.

Fucoxanthin triggers ferroptosis in glioblastoma cells by stabilizing the transferrin receptor.

Glioblastoma (GBM) is the most common and lethal tumor in the world, possessing high stemness, aggressiveness and resistance. Fucoxanthin is a bio-active compound extracted from seaweeds that shows anti-tumor effects to different types of tumors. Here, we show that fucoxanthin inhibits the survival of GBM cells by triggering ferroptosis, a ferric ion and reactive oxygen species (ROS) dependent cell death and ferrostatin-1 could block it. Furthermore, we identified that fucoxanthin targets the transferrin receptor (TFRC). Fucoxanthin is able to block degradation and maintain high levels of TFRC, and similarly inhibits the growth of GBM xenografts in vivo, downregulates the expression of proliferating cell nuclear antigen (PCNA) and upregulates the levels of TFRC in tumor tissues. In conclusion, we demonstrate that fucoxanthin has a significant anti-GBM effect by triggering ferroptosis.

Establishment of a nomogram model to predict the risk of macrosomia in patients with gestational diabetes mellitus.

BACKGROUND: To establish and verify a nomogram model that can predict the risk of macrosomia in patients with gestational diabetes mellitus (GDM). METHODS: Data of patients with GDM who delivered their babies in Shanxi Bethune Hospital between November 2020 and February 2022 were analyzed. Multifactor logistic regression analysis was used to screen the independent risk factors for macrosomia. The model was constructed by R software. The area under the receiver operating characteristic curve (AUC) and goodness-of-fit analysis were used to evaluate its efficiency and accuracy. The clinical application value was evaluated using the decision curve analysis (DCA). RESULTS: A total of 991 patients with GDM were enrolled for modeling. Multigravida, pre-pregnancy body mass index, family history of hypertension, abdominal circumference, and biparietal diameter were independent risk factors for macrosomia, and the prediction model was established. The AUC in the training and test set were 0.93 (0.89-0.97) and 0.90 (0.84-0.96), respectively, and the difference was not statistically significant. The DCA suggested that the model has a high clinical application value. CONCLUSION: The nomogram model for predicting macrosomia in patients with GDM was established. The model has certain accuracy and is expected to be a quantitative tool to guide clinical decision of delivery timing, individualized labor monitoring, and delivery mode.

Investigating the molecular weight distribution of atmospheric water-soluble brown carbon using high-performance size exclusion chromatography coupled with diode array and fluorescence detectors.

Atmospheric brown carbon (BrC) contain amounts of organic species, but their molecular weight (MW) distributions is still poorly understood. This study applied high-performance size exclusion chromatography (HPSEC) coupled with a diode array detector (DAD) and fluorescence detector (FLD) to characterize the MW distributions of typical chromophores and fluorophores within water-soluble BrC. The investigation focused on the spring season, encompassing both typical urban and rural aerosols. Our results showed that chromophores (at 254 and 365 nm), and humic-like and protein-like fluorophores identified by excitation-emission matrix parallel factor analysis (EEM-PARAFAC) within BrC were broadly distributed along the MW continuum (∼50-20,000 Da). This suggests that BrC mainly comprises complex chromophores and fluorophores with heterogeneous molecular sizes. High-MW (HMW, >1 kDa) species (66%-74%) dominated the chromophores at 254 and 365 nm. However, the latter chromophores were enriched with more HMW species. This result suggested that the HMW chromophores might contribute more to BrC absorption at longer wavelengths. The PARAFAC-derived fluorescent components also exhibited different MW distributions. Three humic-like substances (HULIS) were all dominated by HMW fractions (51%-74%), but protein-like fluorescent component (PLOM) enriched low-MW (LMW, <1 kDa) species (60%-66%). Furthermore, the molecular size (i.e., weight-averaged and number-averaged MW) and the ratios between HMW and LMW species decreased in the order highly-oxygenated HULIS > less-oxygenated HULIS > PLOM, indicating that the fluorophores with longer Em were generally related to larger MW. To our knowledge, this is the first report on the molecular size of individual fluorescent components within aerosol BrC. The results obtained here enhanced our knowledge of heterogeneous composition, complex physicochemical properties, and potential atmospheric fates of aerosol BrC.

Novel insights into the angiogenic function of JMJD2B in diabetic hind limb ischemia: involvement of activating Wnt/ß-catenin pathway.

Critical limb ischemia (CLI) is a major health problem, in which diabetes is a risk factor. Lysine Demethylase 4B (JMJD2B) is a histone demethylase. Diabetic CLI model was established in mice by streptozotocin injection and femoral artery ligation. Reduced expression of JMJD2B in lower limb muscles was observed in CLI mice with or without diabetes, accompanied by impaired blood perfusion and mobility. Adenovirus-mediated JMJD2B overexpression improved blood perfusion and angiogenesis as indicated by the alternation in CD31, α-SMA, and VEGFA expression in the lower limb of diabetic mice with CLI. In vitro, JMJD2B expression and the proliferation and tube formation ability were inhibited by high glucose and ischemic conditions in HMEC-1 cells. Overexpressed-JMJD2B contributed to angiogenesis by promoting cell proliferation, migration, and tube formation of HMEC-1 cells, as well as increasing VEGFA and SDF-1 expression. Mechanism study indicated that JMJD2B overexpression activated the Wnt/ß-catenin pathway by promoting ß-catenin nuclear translocation and the expression. This might lead to stimulated angiogenesis, as demonstrated by the Wnt/ß-catenin inhibitor XAV-939. Overall, our study revealed that JMJD2B was down-regulated in CLI mice with diabetes and JMJD2B overexpression promoted angiogenesis probably via the activation of Wnt/ß-catenin pathway.

Quantification of dissipative effects in a complex Ginzburg-Landau equation governed laser system by tracing soliton dynamics.

The concept of dissipative solitons has provided new insight into the complex pulse dynamics in mode-locked lasers and stimulated novel laser cavity designs. However, most of these studies are restricted to qualitative regimes, because it is difficult to quantify dissipative effects in a mode-locked laser. Meanwhile, the quantification of dissipative effects is a general problem that can be also encountered in other dissipative systems. In this paper, we demonstrate a method for quantifying dissipative effects in a mode-locked laser based on analyzing the soliton dynamics traced by time-stretch dispersive Fourier transform. As a result, we are able to quantitatively reproduce the evolution of the pulse that seeds mode-locking through simulations and gain a deeper understanding of the whole process. The obtained physical picture of mode-locking allows us to propose a simple method to quantify the energy threshold for mode-locking buildup and the stability of mode-locked states. A parameter is introduced to evaluate mode-locking conditions, which can serve as a criterion for designing mode-locked lasers. This work opens up new possibilities in the diagnosis and improvement of mode-locked lasers and studies of soliton physics.

Overexpression of the autophagy-related gene SiATG8a from foxtail millet (Setaria italica L.) in transgenic wheat confers tolerance to phosphorus starvation.

In plants, autophagy plays an important role in regulating intracellular degradation and amino acid recycling in response to nutrient starvation, senescence, and other environmental stresses. Foxtail millet (Setaria italica) shows strong resistance to various abiotic stresses; however, current understanding of the regulation network of abiotic stress resistance in foxtail millet remains limited. In this study, we aimed to determine the autophagy-related gene SiATG8a in foxtail millet. We found that SiATG8a was mainly expressed in the stem and was induced by low-phosphorus (LP) stress. Overexpression of SiATG8a in wheat (Triticum aestivum) significantly increased the grain yield and spike number per m2 under LP treatment compared to those in the WT varieties S366 and S4056. There was no significant difference in the grain P content between SiATG8a-overexpressing wheat and WT wheat under normal phosphorus (NP) and LP treatments. However, the phosphorus (P) content in the roots, stems, and leaves of transgenic plants was significantly higher than that in WT plants under NP and LP conditions. Furthermore, the expression of P transporter genes, such as TaPHR1, TaPHR3, TaIPS1, and TaPT9, in SiATG8a-transgenic wheat was higher than that in WT under LP. Collectively, overexpression of SiATG8a increases the P content of roots, stems, and leaves of transgenic wheat under LP conditions by modulating the expression of P-related transporter gene, which may result in increased grain yield; thus, SiATG8a is a candidate gene for generating transgenic wheat with improved tolerance to LP stress in the field.

Foxtail millet MYB-like transcription factor SiMYB16 confers salt tolerance in transgenic rice by regulating phenylpropane pathway.

R2R3-MYB transcription factors play an important role in the synthesis of phenylpropanoid-derived compounds, which in turn provide salt tolerance in plant. In this study, we found that the expression of foxtail millet R2R3-MYB factor SiMYB16 can be induced by salt and drought. SiMYB16 is localized in the nucleus and acts as a transcriptional activator. Phylogenetic analysis indicates that SiMYB16 belongs to the R2R3-MYB transcription factor family subgroup 24. Transgenic rice expressing SiMYB16 (OX16) had a higher survival rate, lower malondialdehyde content, and heavier fresh weight compared with type (WT) under salt stress conditions. The transgenic plants also had a higher germination rate in salt treatment conditions and higher yield in the field compared with wild-type plants. Transcriptome analysis revealed that the up-regulated differential expression genes in the transgenic rice were mainly involved in phenylpropanoid biosynthesis, fatty acid elongation, phenylalanine metabolism, and flavonoid biosynthesis pathways. Quantitative real-time PCR analysis also showed that the genes encoding the major enzymes in the lignin and suberin biosynthesis pathways had higher expression level in SiMYB16 transgenic plants. Correspondingly, the content of flavonoid and lignin, and the activity of fatty acid synthase increased in SiMYB16 transgenic rice compared with wild-type plants under salt stress treatment. These results indicate that SiMYB16 gene can enhance plant salt tolerance by regulating the biosynthesis of lignin and suberin.

Oxygen-evolving hollow polydopamine alleviates tumour hypoxia for enhancing photodynamic therapy in cancer treatment.

Hypoxia, a characteristic hallmark of solid tumours, restricts the therapeutic effect of photodynamic therapy (PDT) for cancer treatment. To address this issue, a facile and nanosized oxygen (O2) bubble template is established by mixing oxygenated water and water-soluble solvents for guiding hollow polydopamine (HPDA) synthesis, and O2 is encapsulated in the cavity of HPDA. HPDA with abundant catechol is designed as a carrier for zinc phthalocyanine (ZnPc, a boronic acid modified photosensitizer) via borate ester bonds to fabricate nanomedicine (denoted as HZNPs). The in vitro and in vivo results indicate that O2-evolving HZNPs could alleviate tumour hypoxia and enhance PDT-anticancer efficiency. Melanin-like HPDA with a photothermal conversion rate (η) of 38.2% shows excellent synergistic photothermal therapy (PTT) efficiency in cancer treatment.

Simple and cost-effective broad bandwidth fiber chirped pulse amplification laser system seeded by a nonlinear amplifier.

In this paper, we demonstrate a simple and cost-effective fiber chirped pulse amplification (CPA) laser system, where a nonlinear amplifier is employed to generate broadband seeding pulses. The nonlinear amplifier can generate stable pulses with 50 nm spectral bandwidth and linear chirp. With such a seeding configuration being adapted into a fiber CPA laser system, the output bandwidth can be expanded from 7 nm to 20 nm, with only minor changes to a standard industrial fiber CPA system. The increased bandwidth allows for pulse durations of less than 100 fs, which is significantly shorter than the original configuration's 250 fs. When combined with a Fourier pulse shaper, such a fiber laser system is expected to produce pulses with energy exceeding 100 µJ and duration shorter than 100 fs.

Effectiveness of Patient-Controlled Intravenous Analgesia (PCIA) with Sufentanil Background Infusion for Post-Cesarean Analgesia: A Randomized Controlled Trial.

Purpose: To investigate the effectiveness of sufentanil patient-controlled intravenous analgesia pump (PCIA) and background infusion in patients of post-cesarean analgesia. Patients and Methods: This trial compared two groups of women undergoing cesarean section and receiving PCIA: no background infusion group (n=30), 6-min lockout time, and background infusion group (n=30), 2 mL/h infusion, 10-min lockout time. Both groups with 2 µg/kg sufentanil was diluted to 100 mL with normal saline. VAS scores at rest at 36 h was the primary endpoint. The secondary endpoints were the VAS scores at rest at 6, 12, and 24 h, the total amount of sufentanil consumed, the Ramsay sedation score (RSS) assessed at the same time points, postpartum bleeding within 24 h, the injection/attempt (I/A) ratio, BP and HR, PONV, side effects of sufentanil. Results: Compared with the no background infusion group, the background infusion group showed lower VAS pain scores at 6, 12, and 24 h (P<0.01), but no differences at 36 h (95% CI = -0.5-0.8. P>0.05). Attempts, injections, and total sufentanil consumption were significantly different between the two groups (P<0.001), but without difference in I/A. Bleeding was less in the background infusion group at 1 h (P=0.03). The minimal respiration rates were not significantly different between groups. Conclusion: Background infusion increased the total consumption of sufentanil within 36 h after cesarean section. Although it did not reduce uterine contraction pain and wound pain at 36 h, it significantly reduced the pain at 6, 12, and 24 h after cesarean section. It improved patient satisfaction and reduced the amount of bleeding after 1 h. Importantly, it did not increase the incidence of hypertension, PONV and respiratory depression.

Energy deposition and incubation effects of nonlinear absorption of ultrashort laser pulses in dielectrics.

Although ultrashort laser has been widely employed in micromachining thanks to its excellent processing precision, one of the main challenges it faces when applied to 3D modification inside dielectrics is its processing efficiency. Many applications require multiple pulses to achieve significant modification to create structure such as microlenses. We report incubation experiments on energy deposition and the control of material modification in fused silica. This allows us to develop a practical incubation model by taking account different ionization mechanisms, in which coefficients relating to multiphoton and avalanche ionization change with laser shots due to accumulating defects. We then extend our study to the scheme where a pre-pulse is used to limit the absorption volume through pre-seeding. Both experiments and simulations show that the efficiency of laser processing can be significantly improved without sacrificing the spatial resolution with this method, especially for longer pulses.