Rate Of Staining Research Articles

Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacteriales.

Infections due to carbapenem-resistant Enterobacteriales, which have increased worldwide in recent years, cause concern. This study aimed to rapidly detect carbapenemase gene region by using flow cytometry in Enterobacteriales isolates and to evaluate its efficiency and susceptibility by comparing it with polymerase chain reaction (PCR). In the study, 21 isolates obtained from the blood cultures of patients hospitalized in intensive care units and found to intermediate or resistant to at least one carbapenem in the automated system, and 14 isolates belonging to the carbapenem-susceptible Enterobacteriales family were included. Carbapenemase gene regions were investigated by PCR after their susceptibility was determined by disk diffusion method. Bacterial suspensions were treated with meropenem + specific carbapenemase inhibitors (EDTA or APBA) and Temocillin and stained with thiazole orange (TO) and propidium iodide (PI) to show dead/live cell differentiation. Dead/live cell percentages were calculated after reading on the flow cytometer device. In the ROC analysis of the flow cytometry method, the cut-off value, specificity, and susceptibility of PI staining rates for meropenem were found as 14.37%, 100%, and 65%, respectively. It was found that the flow cytometry method was well-compatible with PCR in the detection of the carbapenemase gene region. Flow cytometry will continue to be a promising method for the detection of antimicrobial susceptibility and resistance due to its rapid analysis of many cells and its high compatibility with PCR results.

Clinicopathological features and outcome in elderly patients with idiopathic membranous nephropathy

Background Idiopathic membranous nephropathy (IMN) is the leading cause of nephrotic syndrome in the elderly. The treatment of idiopathic membranous nephropathy is quite challenging due to the particularity of elderly patients. This study intends to investigate the clinicopathological characteristics and initial therapeutic effect of idiopathic membranous nephropathy among elderly patients. Methods A retrospective study of 67 elderly patients (58.2% male, median age 69.0 years, range, 65–83 years) with biopsy-proven membranous nephropathy was conducted at Guangdong Provincial People’s Hospital from 2016 to 2020. Data on clinicopathological characteristics and initial therapeutic effects were analyzed. Results Of the 67 patients, the mean eGFR of overall patients was 66.49 mL/min/1.73m2 while the median urine protein-to-creatinine ratio (uPCR) and urine albumin-to-creatinine ratio (uACR) was 5676.73 mg/g and 2951.56 mg/g, respectively. Pathological data revealed that the membranous Churg’s stage II was the most frequent (71.64%). Moreover, glomerular PLA2R antigen fluorescence intensity of (+) and IgG4 antigen fluorescence intensity of (++) were detected in 63.6% and 86.4% of all patients, respectively. Overall, 44 patients, accounting for 65.7%, achieved remission including complete remission and partial remission within 1 year after renal biopsy. Compared with a non-remission group, the levels of uPCR (6274.6 vs. 3235.6 mg/g, p = 0.007) and uACR (3433.6 vs. 1773.2 mg/g, p = 0.017) were significantly higher in remission group. The proportion of immunosuppressive therapy in the remission group was also higher (86.4% vs. 30.4%, p < 0.01). Compared with conservative treatment, patients with combined treatment with glucocorticoid and cyclophosphamide (CTX) or glucocorticoid and calcineurin inhibitor (CNIs) achieved higher remission rate (glucocorticoid plus cyclophosphamide vs. conservative treatment, 84.6% vs. 27.3%, p = 0.001; glucocorticoid plus calcineurin inhibitor vs. conservative treatment, 88.0% vs. 27.3%, p < 0.001). Further analysis showed that compared with patients who underwent conservative treatment, the proportion of males, the levels of uPCR, uACR, BUN, Scr, CysC and PLA2R antigen-positive staining rate in kidney biopsy were higher in those who underwent combined treatment with glucocorticoid and CTX, while the levels of eGFR, TP and ALB were lower (p < 0.05). In addition, patients who received combined treatment with glucocorticoid and CNIs had higher levels of uPCR, uACR, TC and lower levels of TP, ALB than those who received conservative treatment (p < 0.05). Moreover, there were no statistically significant differences in the 1-year progression rate in eGFR between the immunosuppressive treatment group and conservative treatment group (3.3 vs. 0.2 ml/min/1.73m2, p = 0.852). Conclusions Most elderly patients diagnosed with IMN had multiple comorbidities, and the membranous Churg’s stage II was most common. The glomerular PLA2R and IgG4 antigen deposition were frequently detected accompanied by glomerulosclerosis and severe tubulointerstitial injury. For high-risk elderly patients with severe proteinuria, early initial immunosuppressive therapy could achieve a higher urinary protein remission rate. Therefore, it is crucial for clinicians to balance the risks and benefits of immunosuppressive therapy based on clinical and pathological characteristics and develop individualized treatment regimens for elderly patients with IMN.

Luteinizing hormone secretion, ovulatory capacity, and oocyte quality in peripubertal Gir heifers.

Induction of puberty in cattle breeds that attain puberty in later stages, such as Gir, allows the earlier beginning of reproductive life and it might increase oocyte quality. Here, the ovulatory capacity of prepuberal Gir heifers was studied and its relationship to follicular growth, luteinizing hormone (LH) secretion and oocyte quality was evaluated. Peripubertal Gir heifers were treated with a progesterone-based protocol and according to ovulatory response were separated into groups: not-ovulated (N-OV) and ovulated (OV). Serial blood samples were taken 24 h after estradiol treatment on day 12 to evaluate LH secretion. Cumulus-oocyte complexes (COCs) were collected using ovum pick-up and assessed for brilliant cresyl blue (BCB) staining rate, IVF-grade oocytes rate, and mean oocyte diameter, in comparison with cow oocytes. Gene expression of developmental competence markers (ZAR1, MATER, and IGF2R) was also analyzed. The largest follicle diameters were similar between N-OV and OV groups on the day of estradiol treatment (d12) and the next day and decreased (P = 0.04) in the N-OV group thereafter. LH pulse secretion was different between groups (N-OV = 3.61 ± 0.34 vs OV = 2.83 ± 0.21 ng/ ml; P = 0.04). COC assessment showed that the number of recovered oocytes, BCB+ rate, IVF-grade oocytes and oocyte size was similar (P > 0.05) among groups, resembling adult cow patterns. ZAR1, MATER and IGF2R gene expression in oocytes were also similar (P > 0.05) in N-OV and OV groups. In conclusion, our results demonstrate a lower LH secretion profile in peripubertal Gir heifers prone to ovulate after induction protocol, and that oocyte quality is not affected on a short-term basis by ovulation itself.

A Stable and Efficient Genetic Transformation Method for Embryogenic Callus of Fraxinus mandshurica

Fraxinus mandshurica is a widely used greening and ornamental tree species. However, its genetic transformation system has been hampered by problems such as low transformation efficiency, among others, which can hinder research related to molecular breeding and the analysis of functional genes. Thus, in this study, a novel genetic transformation method for efficient transformation of the embryonic callus of Fraxinus mandshurica was investigated. The method was optimized in terms of factors such as antibiotics, infection solution concentrations, co-culture time, and somatic embryo maturation. The results indicated that the optimal antibiotic concentration was 10 mg·L−1 of hygromycin (Hyg). At this point, the callus proliferation multiple was only 0.12. The highest transformation efficiency was found to be 93.93% when the absorbance of the infection solution concentration at OD600 was 0.4. Interestingly, transformation efficiency was found to be highest (77.9%) at 48 h of co-culture, with a GUS staining rate of 88.23%. The medium for somatic embryo maturation of transformed callus was half-strength MS medium (MS 1/2) containing 60 g·L−1 polyethylene glycol, 1 mg·L−1 abscisic acid, 400 mg·L−1 casein enzymatic hydrolysate (CH), 20 g·L−1 sucrose, 1 g·L−1 activated charcoal, and 5 g·L−1 gellan gum. The medium for somatic embryo germination was MS ½, containing 0.2 mg·L−1 of N-(Phenylmethyl)-9H-purin-6-amine(6-BA) and 5.0 mg·L−1 of gibberellin (GA). These results are of significance for the verification of the gene function and future genetic improvement of Fraxinus mandshurica.

The critical role of pancreatic stone protein/regenerating protein in sepsis-related multiorgan failure.

Multiple organ dysfunction syndrome (MODS) is common in patients with sepsistic admitted to an intensive care unit (ICU) and greatly increases mortality. Pancreatic stone protein/regenerating protein (PSP/Reg) is a type of C-type lectin protein that is overexpressed during sepsis. This study aimed to evaluate the potential involvement of PSP/Reg in MODS development in patients with sepsis. The relationship between circulating PSP/Reg levels, patient prognosis, and progression to MODS was analyzed in patients with sepsis admitted to the ICU of a general tertiary hospital. Furthermore, to examine the potential involvement of PSP/Reg in sepsis-induced MODS, a septic mouse model was established per the cecal ligation and puncture procedure, randomized into three groups, and subjected to a caudal vein injection of recombinant PSP/Reg at two different doses and phosphate-buffered saline. Survival analyses and disease severity scoring were performed to evaluate the survival status of the mice; enzyme-linked immunosorbent assays were performed to detect the levels of inflammatory factors and organ-damage markers in murine peripheral blood; terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was performed to measure apoptosis levels in lung, heart, liver, and kidney tissue sections and to visualize the degree of organ damage in the mouse model; myeloperoxidase activity assay, immunofluorescence staining, and flow cytometry were performed to detect neutrophil infiltration levels in vital murine organs and the activation indexes of neutrophils. Our findings indicated that Circulating PSP/Reg levels were correlated with patient prognosis and sequential organ failure assessment scores. Furthermore, PSP/Reg administration increased disease severity scores, shortened survival time, increased the TUNEL-positive staining rate, and increased the levels of inflammatory factors, organ-damage markers, and neutrophil infiltration in the organs. Neutrophils can be activated by PSP/Reg to an inflammatory state, both in vivo and in vitro, which is characterized by the increased levels of intercellular adhesion molecule 1 and CD29. Patient prognosis and progression to MODS can be visualized by monitoring PSP/Reg levels upon ICU admission. Additionally, PSP/Reg administration in animal models exacerbates the inflammatory response and severity of multiorgan damage, which may be accomplished by promoting the inflammatory state of neutrophils.

Systemic antibiotic treatment for meibomian gland dysfunction-A systematic review and meta-analysis.

To review the efficacy and safety of oral doxycycline antibiotics versus macrolides in the treatment of meibomian gland dysfunction (MGD). Systematic review and meta-analysis. We performed a systematic search of electronic databases for all peer-reviewed published studies which included clinical outcomes of oral antibiotic MGD treatment. Individual study data were extracted and evaluated in a weighted pooled analysis, including total sign and symptom scores, meibomian gland secretion score, tear break-up time (TBUT), fluorescein staining score and rate of complications. Two thousand nine hundred and thirty-three studies were found, of which 54 were eligible for the systematic review, and six prospective studies were ultimately included for analysis, reporting on 563 cases from three countries. Age of affected patients ranged between 12 and 90 years. Overall, both treatment methods induced improvement in MGD signs and symptoms. In pooled analysis, macrolides were significantly superior in the total signs score (pooled standardized mean difference (SMD) -0.51, 95% confidence interval (CI): -0.99 to -0.03), meibomian gland secretion score (pooled SMD -0.25, 95%CI: [-0.48, -0.03]), TBUT (SMD -0.31, 95%CI: [-0.50, -0.13]) and fluorescein staining score (SMD -1.01, 95%CI: [-1.72, -0.29]). Moreover, while no severe complications were reported for both treatments, the macrolide group exhibited significantly less adverse events (pooled odds ratio 0.24 with a 95% CI of 0.16 to 0.34). Both macrolides and tetracyclines are effective treatments for MGD. In this study, macrolides exhibited better efficacy and safety profile compared to tetracyclines.

Endometrial Staining of CD56 (Uterine Natural Killer), BCL-6, and CD138 (Plasma Cells) Improve Diagnosis and Clinical Pregnancy Outcomes in Unexplained Infertility and Recurrent IVF Failures: Standardization of Diagnosis with Digital Pathology.

In women with unexplained infertility (UI) and recurrent in vitro fertilization (IVF) failures, the etiology is often unclear. Endometrial immune perturbations and the use of immune markers associated with these dysregulations are of great interest in the diagnosis and treatment of UI. However, reliable biomarkers and standardized quantification methods are lacking. Here, to address endometrial immune dysregulation in UI patients with recurrent IVF failures, we performed endometrial tissue sampling and immunostaining of CD56 (uNK), CD138, and BCL-6. Of these cases, 57.9% had positive CD56 in the endometrial stroma, while 46.1% had positive BCL-6 in the glandular epithelium, and 14.5% of the cases were found to be positive for CD138. Combined staining rates were 60.5%, 68.4%, and 71.05% for (CD56 or BCL-6), (CD56 or CD138), and (CD56, BCL-6, or CD138), respectively. There was a significant correlation between CD56 and BCL-6 positivity, while CD138 positivity was an independent parameter. After the recommended targeted therapy, pregnancy rates were found to increase from 58.5% to 61.6% and 73.8% in CD56-positive, (CD56- or BCL-6-positive), and (CD56-, BCL-6-, or CD138-positive) cases, respectively. Notably, a retrospective evaluation of digital pathology and light microscopy results showed a significant correlation. This study suggests that the examination of CD56, BCL-6, and CD138 in the same endometrial sample may be an effective method in determining the etiology of UI and reaching an early diagnosis and treatment options. Moreover, digital pathology can be used in the evaluation of CD56 and BCL-6 to provide objective, rapid, and reliable results.

Assessment of the efficacy and safety of carbon nanoparticles-guided lymph node dissection in gastric cancer surgery: a systematic review and meta-analysis.

To investigate the efficacy and safety of lymph nodes (LNs) dissection guided by carbon nanoparticles (CNs) in gastric cancer (GC) surgery. We searched electronic databases such as PubMed, Web of Science, Embase, Cochrane Library, and Scopus for relevant articles up to September 2022 and collected all studies comparing the CNs group with blank controls group on the efficacy and safety of LN dissection in gastrectomy. A pooled analysis of the collected data was performed, including the number of retrieved LNs, the staining rate of LNs, the number of metastatic LNs dissection, various intraoperative outcomes, and postoperative complications. A total of 9 studies including 1770 participants (502 in the CNs group and 1268 in the control group) were included. As compared to the blank control group, the CNs group detected 10.46 more LNs in each patient (WMD = 10.46, 95% CI: 6.63 ~ 14.28, p < 0.00001, I2 = 91%), and also significantly more metastatic LNs (WMD = 2.63, 95% CI: 1.43 ~ 3.83, p < 0.0001, I2 = 41%). However, there was no significant difference in the rate of metastatic LNs between the CNs and control groups (OR = 1.37, 95% CI: 0.94 ~ 2.00, P = 0.1, I2 = 89%). In addition, there was no increase in operative time, intraoperative blood loss, and postoperative complications associated with CNs-guided gastrectomy. CNs-guided gastrectomy is safe and effective, and can increase the efficiency of LN dissection without increasing the risk of surgery.

Abstract 4824: Intracellular ferritin expression levels regulate growth and resistance to ferroptosis in pancreatic cancer cell lines

Abstract Background: Pancreatic cancer is one of the cancers with the highest mortality rate, and it is important to develop effective treatments and elucidate prognostic factors. It has been reported that elevated serum ferritin is associated with the prognosis of pancreatic cancer. However, there are few reports on how ferritin in pancreatic cancer tissue acts in pancreatic cancer. Ferritin heavy chain (FTH) is a major component of ferritin, which stores iron and plays a role in suppressing iron-induced generation of reactive oxygen species. In this study, we aimed to examine the expression level of FTH in pancreatic cancer cells and its prognostic and cell biological roles. Methods: We performed immunostaining of FTH in pancreatic cancer tissue specimens operated on at our hospital and classified the expression level of FTH by scoring the immunostaining intensity and staining rate. The expression levels and prognosis were then evaluated. The overexpression and knockout strains of FTH were generated using a human pancreatic cancer cell line (MIAPaCa-2), and their proliferative and stress tolerance abilities were compared with those of the control group. Results: Immunostaining of 149 pancreatic cancer surgical specimens showed that the group with high FTH had a significantly shorter overall survival rate than the group with low FTH (5-year survival rate; 16.8% vs. 36.9%, p = 0.018). In multivariate analysis, high FTH expression was also a prognostic factor: in vitro, cell lines overexpressing FTH had enhanced proliferative potential compared to controls, while knockout lines were suppressed. In addition, the knockout lines showed decreased resistance to RSL-3, an inducer of ferroptosis. Additional studies are currently underway to examine resistance to other drugs and the production of reactive oxygen species in the cells. Conclusion: In pancreatic cancer, high ferritin expression is associated with worse prognosis and may enhance cell proliferation and stress tolerance, and might be a new treatment or biomarker. Citation Format: Masataka Maruno, Yo-hei Kanamori, Takashi Matsumoto, Yuta Shiraishi, Toru Takematsu, Tatsunori Miyata, Kosuke Mima, Shigeki Nakagawa, Hidetoshi Nitta, Hiromitsu Hayashi, Toshiro Moroishi, Hideo Baba. Intracellular ferritin expression levels regulate growth and resistance to ferroptosis in pancreatic cancer cell lines. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4824.

Evaluation of the Presence and Importance of PD-L1 Expression in Head-neck Squamosus Cell Carcinomas and Premalign Lesions

Background: Programmed Death-Ligand 1 (PD-L1) is a cell membrane protein found on the surface of cancer cells, immune system cells in the tumor microenvironment, and various healthy tissues. Moreover, it plays a key role in suppressing the immune system. Objectives: We aim to explain the presence of PD-L1 in Head-Neck Squamosus Cell Carcinomas (HNSCC) and premalign lesions by immunohistochemical method. Methods: Our retrospective study included 22 patients with HNSCC (15(68.1%) Oral cavity and 7(31.9%) Oropharynx), 20 patients with oral lichen planus, and 14 patients with normal oral cavity mucosa. In the evaluation of PD-L1 antibodies applied immunohistochemically in patients with HNSCC, the percentage of tumor cells showing membranous staining with PD-L1 antibodies was calculated. Results: The mean age of HNSCC patients participating in the study was 52.24 ± 11.7 years, the mean age of oral lichen planus patients was 34.10 ± 9.8 years, and the control patients' mean age was 31.42 ± 10.6 years. The rate of PD-L1 staining of tumor cells of HNSCC patients was significantly higher than the control group (P=0.001). Conclusion: The importance of PD-L1 expression in HNSCC and precancerous lesions of the oral cavity is remarkable, and the values may be related to the pathophysiology of these diseases.

Is Orthokeratology Treatment Zone Decentration Effective and Safe in Controlling Myopic Progression?

To compare the myopia control efficacy and safety of decentered versus centered positioning of orthokeratology. This is a retrospective intrasubject study, including 46 children with myopia (25 boys, 21 girls; age 11.12±0.33 years) treated for 1 year with OK decentration in one eye (group D) and central location in the other (Group C). Axial length was measured before and at 6 months and 12 months after the initial lens wear, respectively. Corneal topography was measured at baseline and at 1-month after lens wear. The corneal topography obtained from the 1-month visit was used to quantify treatment zone decentration (TZD) for each subject. Cycloplegic refraction was required for all children before fitting the orthokeratology lenses. No differences were found between the groups in the biological ocular parameters ( P ≥0.05 for all). The axial elongation in group D and group C differed after 6 and 12 months ( P <0.001 for all). Similar corneal staining rates ( P =0.06) were noted during follow-up in groups D (n=20; 7.24%) and C (n=10; 3.62%), all of grade I. The uncorrected visual acuity (UCVA) in group D and C differed after 1, 6, and 12 months ( P =0.002, 0.010, 0.044), except 3 months ( P =0.146). Group D (n=32; 17.39%) was more likely to have glare or ghosting (chi-squared test, P <0.001) than group C (n=12; 6.52%) during follow-up visits. Axial elongation was significantly associated with baseline spherical equivalent (SE) in group C ( P =0.019). In group D, axial elongation was significantly associated with SE and TZD ( P <0.05 for all). This intrasubject study showed that when the UCVA was acceptable and there were no apparent complications, orthokeratology decentration may be beneficial in controlling the progression of myopia. Axial elongation became slower in children with a higher SE and a larger TZD, because TZD ranged from 0.5 mm to 1.5 mm.

Ferric Chloride Controls Citrus Anthracnose by Inducing the Autophagy Activity of Colletotrichum gloeosporioides

Colletotrichum gloeosporioides causes citrus anthracnose, which seriously endangers the pre-harvest production and post-harvest storage of citrus due to its devastating effects on fruit quality, shelf life, and profits. However, although some chemical agents have been proven to effectively control this plant disease, little to no efforts have been made to identify effective and safe anti-anthracnose alternatives. Therefore, this study assessed and verified the inhibitory effect of ferric chloride (FeCl3) against C. gloeosporioides. Our findings demonstrated that FeCl3 could effectively inhibit C. gloeosporioides spore germination. After FeCl3 treatment, the germination rate of the spores in the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) groups decreased by 84.04% and 89.0%, respectively. Additionally, FeCl3 could effectively inhibit the pathogenicity of C. gloeosporioides in vivo. Optical microscopy (OM) and scanning electron microscopy (SEM) analyses demonstrated the occurrence of wrinkled and atrophic mycelia. Moreover, FeCl3 induced autophagosome formation in the test pathogen, as confirmed by transmission electron microscopy (TEM) and monodansylcadaverine (MDC) staining. Additionally, a positive correlation was identified between the FeCl3 concentration and the damage rate of the fungal sporophyte cell membrane, as the staining rates of the control (untreated), 1/2 MIC, and MIC FeCl3 treatment groups were 1.87%, 6.52%, and 18.15%, respectively. Furthermore, the ROS content in sporophyte cells increased by 3.6%, 29.27%, and 52.33% in the control, 1/2 MIC, and MIC FeCl3 groups, respectively. Therefore, FeCl3 could reduce the virulence and pathogenicity of C. gloeosporioides. Finally, FeCl3-handled citrus fruit exhibited similar physiological qualities to water-handled fruit. The results show that FeCl3 may prove to be a good substitute for the treatment of citrus anthracnose in the future.

2,2',4,4',5,5'-Hexabromophenyl ether (BDE-153) causes abnormal insulin secretion and disorders of glucose and lipid metabolism in mice.

Environmental polybrominated diphenyl ether (PBDE) exposure may be associated with diabetes and obesity. 2,2',4,4',5,5'-Hexabromodiphenyl ether (BDE-153) is one of the most abundant and widely distributed homologs of PBDEs detected in humans. This study investigated the effects of BDE-153 on the expression of adipokines and glucose and lipid metabolism. Adult male C57BL/6 mice were divided into five BDE-153 groups and one control group. After BDE-153 exposure for 4 weeks, the levels of biochemical indexes and the mRNA and protein expression levels of leptin, adiponectin, peroxisome proliferators activated receptors gamma (PPARγ), and AMPKα were measured. The histomorphological changes of liver and pancreas tissues were observed. After BDE-153 exposure, the weight of mice in the medium-high-dose group at different exposure times was lower than that in the control group ( p all <0.05), and the body weight decreased slightly with the increase of the dose of BDE-153. BDE-153 caused the disorder of glucose and lipid metabolism in mice, the weight of liver and pancreas increased, lipid droplets accumulated in liver cells, and the positive rate of insulin staining increased in a dose-dependent manner. BDE-153 also interfered with the expression of PPARγ, AMPKα, and adipokines. The results of restrictive cubic splines (RCS) showed that there were a nonlinear dose-response relationship between the exposure dose of BDE-153 and the expression levels of PPARγ, AMPKα, and adipokines. Our results suggest that BDE-153 may interfere with the expression of adipokines and the secretion of insulin by affecting the expression of PPARγ and AMPKα, which play a key role in glucose and lipid metabolism, leading to the occurrence of glucose and lipid metabolism disorder.

Effects of NaOH/H2O2/Na2SiO3 Bleaching Pretreatment Method on Wood Dyeing Properties

Bleaching is a common modification method widely used in the industrialization of wood dyeing. Bleaching can regulate the color of the wood, and it also has great effects on the subsequent wood dyeing properties. In this work, three woods, Ayous, Linden, and Poplar, were processed by using the NaOH/H2O2/Na2SiO3 bleach pretreatment method. Then, the pretreated wood and untreated wood were stained separately by means of water bath dyeing with three dyestuffs: blue anthraquinone and red and yellow double-azo dyestuffs. The study mainly focused on the effect of the bleach pretreatment on the color control and dyeing properties of the wood by analyzing the color difference, staining rate, and final dyeing rate of the dyed wood. The results were as follows: For the color difference, the L* and h* values showed increasing tends, while the a*, b*, and c* values showed decreasing trends. For the staining rate, Ayous reached an equilibrium staining rate at 3~4 h, but for the others, this was not obvious. For the dyeing rate, red and blue Linden veneers and blue Ayous veneers were similar to the unbleached ones, while the final dyeing rate of the other veneers was less than that of the original wood, and the staining rate of the Ayous red and yellow veneer, Linden yellow veneer, and Poplar veneer was less than that of the original wood. The final dyeing rate of the Ayous red and yellow veneers, Linden yellow veneer, and Poplar red, yellow and blue veneers decreased by 4.54%, 2.91%, 5.45%, 10.75%, 2.66%, and 9.55%, respectively. In general, the bleach pretreatment made the staining color of the material surface uniform. The dyeing rate increased due to the increase of the veneers’ permeability. Thus, the veneers quickly reached the equilibrium staining rate, but the equilibrium staining rate of some test pieces decreased. This work provides scientific support for the dyeing process.

Establishment of ultrasound-guided stellate ganglion block in rats.

A novel protocol for accurate stellate ganglion block under ultrasound guidance was designed in rats. This technique raises the success rate of stellate ganglion block and reduces the incidence of brachial plexus and vagus nerve block. Fifty-six Sprague-Dawley were randomly divided into an ultrasound-guided group (n = 28) and a blind technique group (n = 28). The rats in the blind technique group were injected with 1.5% lidocaine mixed with methylene blue after signs of brachial plexus stimulation were elicited. The lateral side of the cephalic brachial vein was located under the first rib, where lidocaine was injected into the rats in the ultrasound-guided group. The up-and-down sequential method of Dixon was used to determine the minimum effective volume for stellate ganglion block in rats. Furthermore, we calculated the required operative duration of the two methods and observed the difference in the lidocaine diffusion range between the two groups. The minimum effective volume for stellate ganglion block in the ultrasound-guided group was 0.040 ml, and the 95% CI was 0.026-0.052 ml. In the blind technique group, the minimum effective volume was 0.639 ml, and the 95% CI was 0.490-0.733 ml. Within the 95% CI of the lowest effective volume, the incidence of brachial plexus block as a complication of stellate ganglion block under ultrasound guidance was 10.00%. Stellate ganglion block under ultrasound guidance is more accurate than blind detection, which the incidence of complications of stellate ganglion block under ultrasound guidance was significantly lower than under blind detection; the rate of methylene blue staining in the vagus nerve was significantly lower under ultrasound guidance.

Protective Effect of Buyang Huanwu Decoction on Cerebral Ischemia/Reperfusion Injury by Inhibiting Autophagy Based on AMPK/UlK1 Signaling Pathway

The aim of the paper is to explore whether AMPK/UlK1 signaling pathway is involved in regulating autophagy and alleviating cerebral ischemia/reperfusion (I/R) injury. On this basis, study the regulatory effect of Buyang Huanwu Decoction (BHD) on AMPK/UlK1 signaling pathway, aiming to clarify the molecular mechanism of BHD in alleviating cerebral I/R injury and the protective effect of BHD containing serum (BHDS) on OGD/R neurons. The research group adopted the following methods: Neurons cultured for 7 days in vitro were randomly divided into normal control group, model group (OGD/R), AMPK activator (AICAR) group (3 mmol/L), Buyang Huanwu Decoction medicated serum (BHDS) group (3%), Buyang Huanwu Decoction medicated serum + AMPK activator (BHDS + AICAR) group. The protein expressions of p-AMPK, AMPK, p-UlK1, UlK1, Beclin-1 and LC3B in neurons were detected by Western blot, the fluorescence intensity and apoptosis rate of neuron autophagosome CYTO-ID staining were detected by flow cytometry, the fluorescence intensity and morphological changes of neuron LC3B were detected by immunofluorescence double labeling method, the morphological changes of neurons were observed by inverted microscope, and the survival rate of neurons was detected by CCK-8 method. Neuron OGD/R model and the optimal concentration of BHDS. The neurons in the model group were subjected to hypoxia and glucose deprivation for 1 h, 2 h, 3 h, 4 h and reoxygenation and glucose deprivation for 24 h. The survival rate of neurons and the release rate of l DH to determine whether the model was successfully established. And the results show that BHDS group, the survival rate of OGD/R neurons was significantly increased, and the apoptosis rate was significantly decreased. Among them, 3% BHDS intervention at the same time of hypoxia glucose deprivation for 3 h and reoxygenation glucose for 24 h was the best condition. In BHDS group, the levels of p-AMPK/AMPK, p-UlK1/UlK1, beclin-1, CYTO-ID and LC3B fluorescence intensity, and the transformation from LC3B-Ⅰ to LC3B-Ⅱ of OGD/R neurons were significantly decreased. So we can draw a conclusion that BHD can inhibit autophagy and alleviate cerebral I/R injury through AMPK/UlK1 signaling pathway.

The safety and efficacy of carbon nanoparticle suspension injection versus indocyanine green tracer-guided lymph node dissection during radical gastrectomy (FUTURE-01): A single-center randomized controlled trial protocol.

The use of lymph node (LN) tracers can help obtain a complete dissection of the lymph nodes and increase the detection rate of LNs and metastatic LNs. Carbon nanoparticle suspension injection (CNSI) and indocyanine green (ICG) have been widely used in radical gastrectomy in recent years. Nevertheless, the comparison of their clinical effects has not been studied. The FUTURE-01 trial will be the first randomized, open-label, single-center trial to compare CNSI and ICG. The study started in 2021 and enrolled 96 patients according to a prior sample size calculation. The primary outcome is the number of LNs retrieved. The secondary outcomes are LN staining rate, LN metastasis rate, stained LN metastasis rate, perioperative recovery and survival. By comparing the safety and efficacy of CNSI and ICG tracer-guided LN dissection in patients with gastric cancer, we can determine the most appropriate LN tracer at present. With the help of LN tracers, the operation is simplified, and the prognosis of these patients is improved. Our study is a prospective exploration of the safety, efficacy, and prognosis of CNSI and ICG. https://clinicaltrials.gov/ct2/show/NCT05229874?cond=NCT05229874&draw=2&rank=1, identifier NCT05229874.

The prognostic effect of immunohistochemical staining rates in patients with non-muscle-invasive bladder cancer.

Despite the follow-up protocols developed in non-muscle-invasive bladder cancer patients, progression and recurrence could not be prevented. We aimed to investigate whether proteins such as OCT-4, CD47, p53, Ki-67, and Survivin, which increase in bladder cancer cells, can be used as prognostic markers for patients with non-muscle-invasive bladder cancer. The study included a total of 89 patients with newly diagnosed non-muscle-invasive bladder cancer between January 2015 and December 2020. Levels of OCT-4, CD47, p53, Kİ-67, and Survivin proteins in cancer cells were determined with a semi-quantitative immunohistochemical experiment. Pathological data and survival rates were compared according to the staining rates. Data obtained in the study were analyzed statistically with SPSS 22.0 (SPSS, Chicago, IL, USA). The mean age of the patients was 64.25 ± 9.91 years, and the median follow-up period was 55 months. Recurrence rate was determined to be 36% (n = 32), and the rate of progression at 40.4% (n = 36). The staining rates were stronger for each marker in the progression group and advanced-stage tumors (p < 0.001). The findings of the multivariate analysis carried out as part of the study showed that older age and higher tumor stage were independent risk factors for recurrence-free survival (HR = 1.048 and 7.074, respectively; P = 0.02). Also, higher tumor stages, diameters, and grades were associated with reduced progression-free survival (HR = 0.105, 0.395, 0.225, respectively; P < 0.05). Although immunohistochemical staining rates are promising, it is more appropriate to use tumor characteristics when assessing survival rate in patients with non-muscle-invasive bladder cancer.

Occurrence of Corneal Staining after Cataract Surgery with and without Chitosan-N-Acetylcysteine Eye Drops

Introduction: The objective of this study was to evaluate the prevalence of ocular surface damage assessed by corneal staining scores right after cataract surgery and whether it can be prevented using chitosan-N-acetylcysteine (C-NAC) eye drops. Methods: We included patients scheduled for routine cataract surgery. Each patient was randomly assigned to one of three groups. Patients in group 1 underwent routine cataract surgery with no additional eye drops. In group 2, patients received C-NAC eye drops after cataract surgery, and in group 3, C-NAC was applied both before and after surgery. Both groups continued the treatment once daily for 4 days. Ocular surface alteration was assessed using the National Eye Institute (NEI) score, and the visual analog scale (VAS) was used to evaluate subjective complaints. Results: Thirty-six patients were included in the final analyses. One hour after cataract surgery, a statistically significant increase in corneal fluorescein staining was observed in all groups, which decreased again after 1 week. There was no significant difference between the groups 1 h after cataract surgery, though a tendency toward lower NEI scores was observed during this time point in group 3. Discussion: Cataract surgery induced ocular surface staining and subjective complaints after 1 h. However, the increase in VAS score was small and probably not clinically relevant. The application of perioperative C-NAC eye drops did reduce the rate of corneal staining after cataract surgery in a clinically relevant manner.

Localization of EGFR Mutations in Non-small-cell Lung Cancer Tissues Using Mutation-specific PNA-DNA Probes.

Epidermal growth factor receptor (EGFR) signaling inhibitors are potent therapeutic agents for EGFR-mutant non-small-cell lung cancer, but the effects of such inhibitors on the localization of EGFR mutations in tumor tissues remain to be elucidated. Thus, a simple and efficient technology for the detection of mutations in tumor tissue specimens needs to be developed. Using an EGFR mutation-specific peptide nucleic acid (PNA)-DNA probe, the EGFR mutation-positive part of whole NSCLC tissues was visualized by immunofluorescence. Formalin-fixed paraffin-embedded sections obtained from A549, NCI-H1975, HCC827 and PC-9 tumors transplanted into nude mice were subjected to staining using PNA-DNA probes specific for the mRNA sequences producing the L858R, del E746-A750 and T790M mutations. The probes for the L858R mutation showed intense positive staining in H1975 cells, and the probe for the del E746-A750 mutation exhibited positive staining specifically in HCC827 and PC-9 tumors. On the other hand, A549 tumors without EGFR mutation did not show any significant staining for any PNA-DNA probe. In combination staining, the addition of cytokeratin stain increased the positive staining rate of each PNA-DNA probe. In addition, the positive staining rate of the probes for the L858R mutation was comparable to that of the antibody to EGFR L858R mutated protein. PNA-DNA probes specific for EGFR mutations might be useful tools to detect heterogeneous mutant EGFR expression in cancer tissues and efficiently evaluate the effect of EGFR signaling inhibitors on tissues of EGFR-mutant cancer.