Effects of Natural Products on Enzymes Involved in Ferroptosis: Regulation and Implications.

Ferroptosis is a form of regulated cell death that is characterized by the accumulation of iron-dependent lipid peroxides. The regulation of ferroptosis involves both non-enzymatic reactions and enzymatic mechanisms. Natural products have demonstrated potential effects on various enzymes, including GPX4, HO-1, NQO1, NOX4, GCLC, and GCLM, which are mainly involved in glutathione metabolic pathway or oxidative stress regulation, and ACSL3 and ACSL4, which mainly participate in lipid metabolism, thereby influencing the regulation of ferroptosis. In this review, we have provided a comprehensive overview of the existing literature pertaining to the effects of natural products on enzymes involved in ferroptosis and discussed their potential implications for the prevention and treatment of ferroptosis-related diseases. We also highlight the potential challenge that the majority of research has concentrated on investigating the impact of natural products on the expression of enzymes involving ferroptosis while limited attention is given to the regulation of enzyme activity. This observation underscores the considerable potential and scope for exploring the influence of natural products on enzyme activity.

Serum zinc-α2-glycoprotein levels are elevated and correlated with thyroid hormone in newly diagnosed hyperthyroidism.

BACKGROUND: Zinc-α2-glycoprotein (ZAG) is a recently novel lipolytic adipokine implicated in regulation of glucose and lipid metabolism in many metabolic disorders. In vitro and animal studies suggest that thyroid hormones (TH) up-regulates ZAG production in hepatocytes. However, there is no data evaluating the possible relationship between ZAG and TH in a human model of hyperthyroidism. The objective of the present study is to assess the association of serum ZAG levels with TH and lipid profile in patients with hyperthyroidism before and after methimazole treatment. METHODS: A total of 120 newly diagnosed overt hyperthyroidism and 122 healthy control subjects were recruited. Of them, 39 hyperthyroidism patients were assigned to receive methimazole treatment as follow-up study for 2 months. RESULTS: The clinical consequence showed that serum ZAG levels were elevated in patients with hyperthyroidism (P < 0.01). Adjust for age, gender and BMI, serum ZAG levels were positively related with serum free T3 (FT3), free T4 (FT4) levels and negatively correlated with serum total cholesterol (TC), low density lipoprotein cholesterol (LDLC) levels in hyperthyroidism subjects (all P < 0.01). After methimazole treatment, serum ZAG levels were decreased and the decline was associated with decreased FT3, FT4 and increased TC levels (all P < 0.001). CONCLUSION: We conclude that ZAG may be involved in the pathogenesis of lipid metabolism disorder in patients with hyperthyroidism. TRIAL REGISTRATION: ChiCTR-ROC-17012943 . Registered 11 October 2017, retrospectively registered.

[Ultrasound characteristics of testicular Leydig cell tumors , , , 1.]

OBJECTIVE: To explore the color Doppler ultrasonic characteristics of testicular Leydig cell tumors (LCT) and improve the clinical diagnosis of the disease. METHODS: We retrospectively analyzed 4 cases of testicular LCT diagnosed and treated in our hospital and summarized the experience in the ultrasonic diagnosis of LCT with a review of the relevant literature. RESULTS: All the 4 testicular LCTs were solitary and quasi-round, 1 in the left and 3 in the right. The smallest mass was 1.8 × 1.5 cm and the largest 3.1 × 2.5 cm, and 2 were complicated by hydrocele of tunica vaginalis. The margins of tumors were distinct in 2 cases and indistinct in 1, and changed from distinct to indistinct in another during the follow-up. Hypoechoes were revealed in all the 4 cases in ultrasonography, 2 with abundant internal blood flow, 1 with abundant peripheral blood flow, and the other with abundant internal blood flow changed from circular blood flow surrounding the mass. CONCLUSIONS: A typical sporadic LCT was ultrasonically manifested as an isolated hypoechoic infracentimetric mass with a clear demarcation from the adjacent pulp. It exhibited intrinsic hypervascularization associated with a typical peripheral rim pattern. Larger lesions more often presented a lobulated shape and intense hypervascularization. Although these ultrasonic characteristics do not reveal the nature of LCT with certainty, they can help the surgeon with the decision on testis-sparing surgery or perhaps even on the active monitoring for the smallest lesions in a population with impaired fertility.

Nrf2 protects human lens epithelial cells against H2O2-induced oxidative and ER stress: The ATF4 may be involved.

Our previous study has shown heme oxygenase-1 (HO-1) protects human lens epithelial cells (LECs) against H2O2-induced oxidative stress and apoptosis. Nrf2, the major regulator of HO-1, is triggered during the mutual induction of oxidative stress and ER stress. In response to ER stress, unfolded protein response (UPR) serves as a program of transcriptional and translational regulation mechanism with PERK involved. Both Nrf2 and ATF4 are activated as the downstream effect of PERK signaling coordinating the convergence of dual stresses. However, the ways in which Nrf2 interacting with ATF4 regulates deteriorated redox state have not yet been fully explored. Here, the transfected LECs with Nrf2 overexpression illustrated enhanced resistance in morphology and viability upon H2O2 treatment condition. Intracellular ROS accumulation arouses ER stress, initiating PERK dependent UPR and inducing the downstream signal Nrf2 and ATF4 auto-phosphorylation. Further, converging at target promoters, ATF4 facilitates Nrf2 with the expression of ARE-dependent phase II antioxidant and detoxification enzymes. According to either Nrf2 or ATF4 gene modification, our data suggests a novel interaction between Nrf2 and ATF4 under oxidative and ER stress, thus drives specific enzymatic and non-enzymatic reactions of antioxidant mechanisms maintaining redox homeostasis. Therapies that restoring Nrf2 or ATF4 expression might help to postpone LECs aging and age-related cataract formation.

Untargeted lipidomic analysis and network pharmacology for parthenolide treated papillary thyroid carcinoma cells.

BACKGROUND: With fast rising incidence, papillary thyroid carcinoma (PTC) is the most common head and neck cancer. Parthenolide, isolated from traditional Chinese medicine, inhibits various cancer cells, including PTC cells. The aim was to investigate the lipid profile and lipid changes of PTC cells when treated with parthenolide. METHODS: Comprehensive lipidomic analysis of parthenolide treated PTC cells was conducted using a UHPLC/Q-TOF-MS platform, and the changed lipid profile and specific altered lipid species were explored. Network pharmacology and molecular docking were performed to show the associations among parthenolide, changed lipid species, and potential target genes. RESULTS: With high stability and reproducibility, a total of 34 lipid classes and 1736 lipid species were identified. Lipid class analysis indicated that parthenolide treated PTC cells contained higher levels of fatty acid (FA), cholesterol ester (ChE), simple glc series 3 (CerG3) and lysophosphatidylglycerol (LPG), lower levels of zymosterol (ZyE) and Monogalactosyldiacylglycerol (MGDG) than controlled ones, but with no significant differences. Several specific lipid species were changed significantly in PTC cells treated by parthenolide, including the increasing of phosphatidylcholine (PC) (12:0e/16:0), PC (18:0/20:4), CerG3 (d18:1/24:1), lysophosphatidylethanolamine (LPE) (18:0), phosphatidylinositol (PI) (19:0/20:4), lysophosphatidylcholine (LPC) (28:0), ChE (22:6), and the decreasing of phosphatidylethanolamine (PE) (16:1/17:0), PC (34:1) and PC (16:0p/18:0). Four key targets (PLA2G4A, LCAT, LRAT, and PLA2G2A) were discovered when combining network pharmacology and lipidomics. Among them, PLA2G2A and PLA2G4A were able to bind with parthenolide confirmed by molecular docking. CONCLUSIONS: The changed lipid profile and several significantly altered lipid species of parthenolide treated PTC cells were observed. These altered lipid species, such as PC (34:1), and PC (16:0p/18:0), may be involved in the antitumor mechanisms of parthenolide. PLA2G2A and PLA2G4A may play key roles when parthenolide treated PTC cells.

Unveiling 2,000 years of differentiation among Tungusic-speaking populations: a revised phylogeny of the paternal founder lineage C2a-M48-SK1061.

Previous studies demonstrated Y chromosome haplogroup C2a-M48-SK1061 is the only founding paternal lineage of all Tungusic-speaking populations. To infer the differentiation history of these populations, we studied more sequences and constructed downstream structure of haplogroup C2a-M48-SK1061 with better resolution. In this study, we generated 100 new sequences and co-analyzed 140 sequences of C2a-M48-SK1061 to reconstruct a highly revised phylogenetic tree with age estimates. We also performed the analysis of the geographical distribution and spatial autocorrelation of sub-branches. Dozens of new sub-branches were discovered, many sub-branches were nearly unique for Ewenki, Evens, Oroqen, Xibe, Manchu, Daur, and Mongolian. The topology of these unique sub-branches is the key evidence for understanding the complex evolutionary relationship between different Tungusic-speaking populations. The revised phylogeny provided a clear pattern for the differentiation history of haplogroup C2a-M48-SK1061 in the past 2,000 years. This study showed that the divergence pattern of founder lineage is essential to understanding the differentiation history of populations.

Association of Peripheral Blood Cell Profile With Alzheimer's Disease: A Meta-Analysis.

Background: Inflammation and immune dysfunction play significant roles in the pathogenesis of Alzheimer's disease (AD)-related dementia. Changes in peripheral blood cell profiles are a common manifestation of inflammation and immune dysfunction and have been reported in patients with AD or mild cognitive impairment (MCI). We systematically evaluated the association of peripheral blood cell counts and indices with AD or MCI through a meta-analysis. Methods: We electronically searched sources to identify all case-control trials comparing peripheral blood cell counts and/or lymphocyte subsets between patients with AD or MCI and healthy controls (HCs). Meta-analyses were used to estimate the between-group standardized mean difference (SMD) and 95% confidence interval (CI). Results: A total of 36 studies involving 2,339 AD patients, 608 MCI patients, and 8,352 HCs were included. AD patients had significantly decreased lymphocyte counts (SMD -0.345, 95% CI [-0.545, -0.146], P = 0.001) and significantly increased leukocyte counts (0.140 [0.039, 0.241], P = 0.006), neutrophil counts (0.309 [0.185, 0.434], P = 0.01), and neutrophil-lymphocyte ratio (NLR) (0.644 [0.310, 0.978], P < 0.001) compared to HCs. Similarly, significantly increased leukocyte counts (0.392 [0.206, 0.579], P < 0.001), NLR (0.579 [0.310, 0.847], P < 0.001), and neutrophil counts (0.248 [0.121, 0.376], P < 0.001) were found in MCI patients compared with HCs. A significantly decreased percentage of B lymphocytes (-1.511 [-2.775, -0.248], P = 0.019) and CD8+ T cells (-0.760 [-1.460, -0.061], P = 0.033) and a significantly increased CD4/CD8 ratio (0.615 [0.074, 1.156], P = 0.026) were observed in AD patients compared to HCs. Furthermore, significant changes in hemoglobin level and platelet distribution width were found in patients with AD or MCI compared with HCs. However, no significant difference was found between AD or MCI patients and HCs in terms of platelet counts, mean corpuscular volume, red cell distribution width, mean platelet volume, and CD4+ T, CD3+ T, or natural killer cell counts. Conclusion: Changes in peripheral blood cell profiles, particularly involving leukocyte, lymphocyte, neutrophil, and CD8+ T cell counts, as well as the NLR and the CD4/CD8 ratio, are closely associated with AD. The diagnostic relevance of these profiles should be investigated in future.

The effect of dentin surface treatment with disinfectant on the shear bonding strength of luting cements.

Background/purpose: Few studies have comprehensively assessed the shear bonding strength of the luting cements between abutments and fixed partial dentures after dentin surface treatment with disinfectants. The purpose of the study was to evaluate the effect of three commonly used disinfectants (2.5% sodium hypochlorite, 0.2% chlorhexidine, and 0.2% benzalkonium chloride) on the shear bonding strength of four luting cements. Materials and methods: Teeth were mounted on Teflon cylinders and prepared for dentin exposure. Three different disinfectants were used to treat the dentin surface. Nickel-chromium posts were cemented with resin cement, glass ionomer cement, polycarboxylate cement, or zinc phosphate cement. The shear bonding strength of the cement was examined using an Instron testing machine. Two-way analysis of variance (ANOVA) was used to examine the differences in shear bonding strength between the cements. If a statistically significant difference was found through ANOVA, a post hoc test with Tukey's honest significant difference was conducted. Results: Disinfectants significantly decreased the shear bonding strength of resin cement, with 2.5% sodium hypochlorite causing the most substantial decrease. The zinc phosphate cement group displayed minimal shear bonding strength regardless of the disinfectant used. Conclusion: The presence of 2.5% sodium hypochlorite significantly reduced the shear bonding strength of resin cements. During permanent cementation of indirect restorations, the choice of luting cement paired with the proper disinfectant is of utmost importance to maintain the shear bonding strength.

Expression profiles of tRNA-derived fragments and their potential roles in lung adenocarcinoma.

Background: Transfer RNA-derived fragments (tRFs) and transfer RNA halves (tiRNAs) have been shown to play crucial roles in gene regulation. This study aims to reveal the expression profiles of tRFs and tiRNAs and their possible biological roles in lung adenocarcinoma (LUAD). Methods: Five paired clinical lung adenocarcinoma tissues (LAT) and adjacent normal lung tissues (ANLT) were selected to analyze the expression of tRFs and tiRNAs. Six significantly expressed tRFs and tiRNAs were selected and validated by Quantitative Real-time PCR (qPCR). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed. Results: The sequencing results showed that 109 tRFs and tiRNAs were differentially expressed between LAT and ANLT, out of which 60 were upregulated and 49 were downregulated. Compared with ANLT, lower expression levels of 3 tRF-1s (tRF-Ser-TGA-010, tRF-Arg-CCT-018, and tRF-Val-CAC-017) in LAT were verified by qPCR. Subsequently, the putative target genes of tRF-1s were analyzed by computational prediction and the top 10 significant results of GO and KEGG pathway enrichment analysis were presented. Conclusions: This study has revealed the landscape of tRF and tiRNA expression profiles in LUAD. Three newly found differentially expressed downregulated tRF-1s may be involved in the pathogenesis of LUAD and may serve as potential diagnostic biomarkers, or otherwise reconcile target genes for drug development.

Cell wall composition contributes to the control of transpiration efficiency in Arabidopsis thaliana.

To identify loci in Arabidopsis involved in the control of transpirational water loss and transpiration efficiency (TE) we carried out an infrared thermal imaging-based screen. We report the identification of a new allele of the Arabidopsis CesA7 cellulose synthase locus designated AtCesA7(irx3-5) involved in the control of TE. Leaves of the AtCesA7(irx3-5) mutant are warmer than the wild type (WT). This is due to reduced stomatal pore widths brought about by guard cells that are significantly smaller than the WT. The xylem of the AtCesA7(irx3-5) mutant is also partially collapsed, and we suggest that the small guard cells in the mutant result from decreased water supply to the developing leaf. We used carbon isotope discrimination to show that TE is increased in AtCesA7(irx3-5) when compared with the WT. Our work identifies a new class of genes that affects TE and raises the possibility that other genes involved in cell wall biosynthesis will have an impact on water use efficiency.

Escherichia coli induces apoptosis in human monocytic U937 cells through the Fas/FasL signaling pathway.

Apoptosis is a genetically regulated cellular suicide mechanism that plays an essential role in development and in defense of multicellular organism. Escherichia coli (E. coli) can induce monocyte apoptosis; however, the mechanism is not clear. This study determines if Fas/FasL regulates E. coli-induced human monocyte line U937 cell apoptosis. We found that infection of U937 cells with E. coli induced rapid cell death in a dose- and time-dependent manner displaying the characteristic features of apoptosis. Moreover, opsonized E. coli induced U937 apoptosis with a higher apoptotic rate (53.29 ± 5.83%) than non-opsonized E. coli (19.37 ± 2.56%). Studying the underlying mechanisms we found that the E. coli-induced apoptosis was associated with a more prominent induction expression of Fas/FasL in a time- and dose-dependent manner. Furthermore, E. coli treatment resulted in a significant increase in the levels of DR5, TRAIL, and FADD, but exerted no statistically significant effects on the levels of DR4. The activity of caspase-8 enzyme increased in infection groups, positively correlated with apoptosis rate. Taken together, these results clearly indicate that receptor-mediated phagocytosis of E. coli induces apoptosis. Moreover, our findings suggest a possible regulatory role of Fas/FasL in the pathway of E. coli infection.

[P-selectin gene -2123 polymorphism in children with Henoch-Sch-nlein purpura].

OBJECTIVE: To investigate whether P-selectin gene -2123 polymorphism is associated with the pathogenesis of Henoch-Sch-nlein purpura (HSP) in children. METHODS: Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) is used to identify the distribution of allele and genotype frequencies of P-selectin gene promoter -2123 polymorphism in 86 children with HSP (including 40 cases of purpura nephritis) and 70 healthy controls. RESULTS: Compared with the healthy controls, the frequencies of GG genotype and G allele of P-selectin promoter -2123 in children with HSP increased significantly (P<0.05). There were no significant differences in P-selectin promoter -2123 genotype and allele frequencies between the patients with and without nephritis. CONCLUSIONS: P-selectin gene promoter -2123 polymorphism appears to be associated with the pathogenesis of HSP in children.

Comparison of the internal fit of metal crowns fabricated by traditional casting, computer numerical control milling, and three-dimensional printing.

This experimental study aimed to compare the internal fit (marginal fit and internal discrepancy) of metal crowns fabricated by traditional casting and digital methods (computer numerically controlled (CNC) milling and three-dimensional [3D] printing). Thirty standard master abutment models were fabricated using a 3D printing technique with digital software. Metal crowns were fabricated by traditional casting, CNC milling, and 3D printing. The silicon replica method was used to measure the marginal and internal fit. A thin layer of low-viscosity polyvinyl siloxane material was placed inside each crown and on the die (like a seat) until the material was set. Replicas were examined at four reference points under a microscope: the central pit (M1), cusp tip (M2), axial wall (M3), and margin (M4). The measured data were analyzed using a one-way analysis of variance (ANOVA) to verify statistical significance, which was set at p < 0.05. In the traditional casting group, the minimum distance measured was at M3 (90.68 ± 14.4 µm) and the maximum distance measured was at M1 (145.12 ± 22 µm). In the milling group, the minimum distance measured was at M3 (71.85 ± 23.69 µm) and the maximum distance measured was at M1 (108.68 ± 10.52 µm). In the 3D printing group, the minimum distance measured was at M3 (100.59 ± 9.26 µm) and the maximum distance measured was at M1 (122.33 ± 7.66 µm). The mean discrepancy for the traditional casting, CNC milling, and 3D printing groups was 120.20, 92.15, and 111.85 µm, respectively, showing significant differences (P < 0.05). All three methods of metal crown fabrication, that is, traditional casting, CNC milling, and 3D printing, had values within the clinically acceptable range. The marginal and internal fit of the crown was far superior in the CNC milling method.

Prevalence of congenital dental anomalies in the primary dentition in Taiwan.

PURPOSE: The purpose of this study was to investigate the prevalence of congenital anomolies in the primary dentition in preschool Taiwanese children compared to Caucasion populations. METHODS: The study group comprised 2,611 children (1,442 boys ond 1,169 girls) between 2 and 6 years-old in the primary dentition phase using a random sampling technique. The presence of hypodontia, hyperdontia, and double teeth (fusion ond gemination) were recorded ond analyzed using Fisher's exact test. RESULTS: The prevalence of hypodontia was approximately 2%. Unilaterol missing teeth were more frequently observed than bilateral missing teeth. The primary mandibular right lateral incisor was the most frequently missing tooth. The prevalence af hyperdontia was less than 1%. All of the supernumerary teeth were located in the primary maxillary incisor area. The prevalence of double teeth was approximately 3%. All double teeth were located in the anterior region, and most of them were found in the mandibular lateral incisor and canine area. There were no differences in the prevalence of anomalies between the sexes. CONCLUSIONS: The prevalence of hypodontia and double teeth, but not hyperdontia, was significantly higher in Taiwanese children vs Caucasian children. Early detection of primary dental anomalies may help prevent oral disease in children.

[Origins determination of Herba Abri cantoniensis and Herba Abri mollis with FTIR combined with fuzzy cluster and curve-fitting].

The methods of fuzzy cluster and curve-fitting combined with FTIR were used to determine the origins of Herba Abri cantoniensis and Herba Abri mollis. The spectra of Herba Abri cantoniensis and Herba Abri mollis are similar, both with typical spectral shapes. The two spectra can be divided into 3 parts: the 1st is 3 500-2 800 cm(-1), containing stretching bands of -OH, N-H, and CH2 ; the 2nd is 1 800-800 cm(-1), containing stretching bands of ester carbonyl group and indican C-O(H), vibrational bands of C=C and benzene ring; The 3rd is 800-400 cm(-1), containing skeletal vibration and scissoring vibration of molecular. The recorded FTIR spectral data were processed by 9-point-smoothing, 1st derivative, SNV and fuzzy cluster analysis sequentially. The fuzzy cluster analysis was carried out by similarity or dissimilarity matrix, and two matrices are computed with Manhattan and Euclidean distance. The results indicated that the optimization used Manhattan and dissimilarity matrix, and 5 origins of Herba Abri cantoniensis were perfectly discriminated, but 2 origins of Herba Abri mollis were mixed and identified from the other 3 origins. So the characterized bands at 1 034 cm(-1) of the average 1-D spectra of Herba Abri cantoniensis and Herba Abri mollis were fitted combining 2nd derivative for further distinguishing their spectral characteristic. The results of curve-fitting showed that the bands of wild Herba Abri cantoniensis and the other origin ones were decomposed to 11 and 9 component bands respectively, but the bands of Shanglin and the other origins Herba Abri mollis were decomposed to 9 and 8 component bands dissimilarly, and the locations and normalized densities of these component bands were different. From this, together with the results of fuzzy cluster analysis, it is concluded that the combination of two methods may identify the origins of Herba Abri cantoniensis and Herba Abri mollis availably.

[Comparative study on the infrared fingerprint of Abrus cantoniensis based on the methods of sequential analysis of dual-indexes and cluster analysis].

The methods of sequential analysis of dual-indexes and cluster analysis were utilized to investigate the infrared fingerprints of A. cantoniensis planted in different years and different places in Guangxi, China. The results showed that 6 samples were able to be completely separated only through 13 point smoothing, when the dual-indexes analysis was applied in the present research, and the accurate relationship between these samples could be inspected and expressed by quantitative relationships under 6-dimensional spaces; however, the effect of cluster was bad only through 13 point smoothing of raw spectra, and it was very difficult to find out the regular sequences while the cluster analysis was applied. Furthermore, the 6 samples were able to be completely separated if raw spectra were dealed with 1st derivative after 13 point smoothing, and the clustering effects were more obvious and 6 samples of A. cantoniensis were completely separated. The above two methods could be used to evaluate the quality of Chinese medicinal materials easily when the sample was not excessive quantitatively, but the method of dual-indexes analysis was more difficult than the clustering analysis if the sample size was too large, since a mass of data such as common peak ratio and variation peak ratio of the IR fingerprint spectra were processed and analyzed statistically, while this method could accurately find out the closest relationship between any samples through comparing the quantitative relationships of common peak ratio and variation peak ratio of each sample under 6-dimensional space; the precision of cluster analysis was less than dual-indexes analysis, but it was more convenient than dual-indexes analysis when large sample data were analysed. Finally the above two methods all showed that the chemical composition of the A. cantoniensis was similar in the same cultivated area, but the difference in chemical composition of A. cantoniensis in different years was distinct even they were in the same place.

Primary anorectal malignant melanoma: a case report.

Anorectal melanoma (ARM) is rare and lethal. We report a case of a 48-year-old woman with 9 months of rectal swelling and bleeding. Physical examination revealed a mass about 5 × 6 cm on the anterior wall of the rectum, 3 cm from the anal verge, and the patient underwent abdominoperineal resection (APR). After hematoxylin-eosin staining and immunohistochemical staining, it was considered an ARM, which is an aggressive disease with a poor survival. Immunohistochemical staining showed the tumor to be positive for S-100, Melan A, Ki67 proliferative index of 70%, and negative for HMB45. The melanoma had infiltrated the adventitia and metastasized to the (intestinal) 16/16 lymph nodes with cancerous nodule formation. There were multiple organs with metastasis (liver, spleen, pancreas, lung and subcutaneous soft tissue) three months after operation. Overall, pre-operative biopsy may be insufficient to make a definite diagnosis, and immunohistochemistry is necessary. Therefore, the gold standard treatment for ARM is oncological radical surgical resection.

Benazepril, an angiotensin-converting enzyme inhibitor, alleviates renal injury in spontaneously hypertensive rats by inhibiting advanced glycation end-product-mediated pathways.

1. Advanced glycation end-products (AGE) and their receptors (RAGE) have been implicated in renal damage in diabetes. The aim of the present study was to investigate the effects of benazepril, an angiotensin-converting enzyme inhibitor (ACEI), on the formation of AGE, the expression RAGE and other associated components in the oxidative stress pathway in spontaneously hypertensive rats (SHR). 2. Groups of SHR were treated with or without 10 mg/kg per day benazepril for 12 weeks. Systolic blood pressure (SBP) and angiotensin (Ang) II levels were evaluated in SHR and control Wistar-Kyoto (WKY) rats. Renal function was investigated by determining levels of proteinuria and glomerulosclerosis. Furthermore, reactive oxygen species (ROS) in the rat renal cortex were analysed using an H(2)O(2)-based hydroxyl radical-detection assay and the renal content of AGE, RAGE, NADPH oxidase p47phox, nuclear factor (NF)-kappaB p65, phosphorylated (p-) NF-kappaB p65, vascular cell adhesion molecule (VCAM)-1 and transforming growth factor (TGF)-beta1 was determined by immunohistochemistry, quantitative real-time polymerase chain reaction and western blot analysis. 3. Treatment with benazepril inhibited the formation of AngII, reduced SBP and alleviated renal lesions in SHR compared with both untreated SHR and control WKY rats. Benazepril treatment significantly suppressed the accumulation of AGE and expression of RAGE in the kidney of SHR. In addition, benazepril treatment reduced the upregulation of NADPH oxidase p47phox, ROS generation and NF-kappaB p65, p-NF-kappaB p65, VCAM-1 and TGF-beta1 expression in the kidney of SHR compared with both untreated SHR and control WKY rats. 4. The results of the present study provide new insights into the regulation by the renin-angiotensin system of AGE-RAGE, oxidative stress and nephropathy, increasing our understanding of the role of the RAS in nephropathy.

[Application of carbocyanines fluorescent dye for tracing fetal fibroblast transplant: experiment with rats].

OBJECTIVE: To investigate the feasibility of applying carbocyanines fluorescent dye chloromethyl-benzamidodialkyl carbocyanine (CM-Dil) in tracing fetal fibroblast transplant. METHODS: Sixteen-day-gestation fetal rat skin tissue samples were harvested and digested with trypsin to isolate fibroblasts. The fibroblasts proliferated in logarithmic phase and were incubated with CM-Dil for 15 minutes. Flow cytometry was used to calculate the proliferate index (PI) so as to determine the effect of CM-Dil on cell proliferation. Labeled fibroblasts were transplanted to the deep partial thickness burn wounds in the back of allogenic rats. Samples were harvested on 0, 1, 3, 7, 14, and 28 days post burns (DPB) and observed under fluorescence microscope in 6 microm section. RESULTS: The cultured fibroblasts were labeled with CM-Dil within 15 minutes, appearing as red fluorescent discs with bilayer membrane structure. The PI of the labeled fibroblasts was (76.9 +/- 2.8), not significant different from that of the cells not labeled (75.8 +/- 2.0, t = 0.80, P > 0.05). The area with fluorescence markers was small on 1 DPB, began to expand and be distributed in a beaker shape on 3 DPB, and then the bottom of the "beaker" broadened indicating cell proliferation significantly on 7 DPB. Fluorescence signals determined were moderate on 14 DPB, and can be still determined feebly on 28 DPB. CONCLUSION: Proper concentration of CM-Dil shows no cytotoxicity to fetal fibroblasts. CM-Dil is an ideal fluorescent label to fetal fibroblasts for its fairly stable fluorescent tracing and slow quenching in skin tissue within 4 week's duration.

[Using Raman spectroscopy to analyze apoptosis of gastric cancer cells induced by cisplatin].

Apoptosis of gastric cancer cells induced by cisplatin was investigated using laser Raman spectroscopy. Gastric cancer cells (SGC7901) were treated with 10 microg x mL(-1) cisplatin for 24, 48 and 72 hours, then were divided into two parts, one for fluorescence staining, the other for collection of Raman spectra by means of scanning. The acquired spectra were then preprocessed by background elimination, smoothing, normalization, baseline correction, and peak fitting. Fluorescence staining result showed that the nucleuses from untreated group were uniformly stained, while those from the group treated for 72 hours were densely stained and broken. The spectra results revealed that the intensity of peaks associated with nucleic acid and protein decreased after the cells were incubated with cisplatin for 24, 48 and 72 hours. The intensity of peaks at 783, 1002 and 1343 cm(-1) respectively fell to 52, 64 and 76 percent of the original value after 72 h of treatment, which indicated that cisplatin could induce apoptosis of gastric cancer cells and reduce the amount of nucleic acid and protein in the cells. The above results suggest that Raman spectra can provide abundant information about the changes in materials in cells and serve as an effective method for real time measurement of apoptosis.