CD4+ T cells apoptosis and conversion of Th1/Th2-type cytokines promote the progress of sporotrichosis.

Sporotrichosis is a fungal disease of the human and other mammals, caused by a complex of Sporothrix schenckii. The disease follows the traumatic inoculation to lead to fixed lesions, regional lymphangitic lesions, or even disseminated lesions including internal involvement, which depends on host immunological status and strain virulence. In this work, we observed the role of CD4+ T cells apoptosis and conversion of Th1/Th2-type cytokines in the cellular immunity regulation on mice model sporotrichosis. The experiments showed that there was more CD4+ T cells apoptosis, by endogenous apoptosis signaling pathway (P < .05), and more conversions of Th1/Th2-type cytokines in more severe and longer duration groups (P < .05). Meanwhile, the trends of the conversions of Th1/Th2-type cytokines were almost consistent with the CD4 + T cell's apoptosis in the corresponding groups. These findings suggest that CD4+ T cells apoptosis and conversion of Th1/Th2-type cytokines are contributing to promoting the progress of sporotrichosis.

AgNAC1, a celery transcription factor, related to regulation on lignin biosynthesis and salt tolerance.

The NAC transcription factor participates in various biotic and abiotic stress responses and plays a critical role in plant development. Lignin is a water-insoluble dietary fiber, but it is second only to cellulose in abundance. Celery is the main source of dietary fiber, but its quality and production are limited by various abiotic stresses. Here, AgNAC1 containing the NAM domain was identified from celery. AgNAC1 was found to be a nuclear protein. Transgenic Arabidopsis thaliana plants hosting AgNAC1 have longer root lengths and stomatal axis lengths than the wide type (WT). The evidence from lignin determination and expression levels of lignin-related genes indicated that AgNAC1 plays a vital role in lignin biosynthesis. Furthermore, the results of the physiological characterization and the drought and salt treatments indicate that AgNAC1-overexpressing plants are significantly resistive to salt stress. Under drought and salt treatments, the AgNAC1 transgenic Arabidopsis thaliana plants presented increased superoxide dismutase (SOD) and peroxidase (POD) activities and decreased malondialdehyde (MDA) content and size of stomatal apertures relatively to the WT plants. The AgNAC1 served as a positive regulator in inducing the expression of stress-responsive genes. Overall, the overexpressing AgNAC1 enhanced the plants' resistance to salt stress and played a regulatory role in lignin accumulation.

[Serum exosomes of rats with hepatic ischemia-reperfusion injury induce the damage of hippocampus and cerebral cortex].

The paper was aimed to explore the role of serum exosomes induced by hepatic ischemia/reperfusion (I/R) injury in the damage of hippocampus and cerebral cortex of rats. The male Sprague-Dawley (SD) rats were randomly divided into 4 groups: sham operation group (S), hepatic I/R injury group (I/R), serum exosomes from S group treatment group (ES) and serum exosomes from I/R group treatment group (EI). In ES group and EI group, 100 µL serum exosomes from S group and I/R group were injected into the normal rats through tail vein respectively. Another three normal rats were injected intravenously with serum exosomes labeled with PKH26 red fluorescence, and then the expression of fluorescence in the brain tissues was observed by immunofluorescence microscope. The morphology and size of exosomes were observed by transmission electron microscope, the expression of exosomes markers CD63 and CD9 was detected by Western blot, and the damage of liver and brain, levels of apoptosis and oxidative stress response in hippocampus and cerebral cortex were observed by serological and histological indexes. The results showed that the exosomes were a group of round or ovoid membranous vesicles, sized in 30-100 nm. Compared with that in S group, the content of serum exosomes in I/R group was increased (P < 0.05). Moreover, serum exosomes could go through the blood-brain barrier and enter the brain tissue freely through blood circulation. The index of liver function in I/R group was significantly higher than that in S group (P < 0.05). There was no significance in the degree of brain damage, apoptosis and oxidative stress in hippocampus and cerebral cortex between S group and ES group. Compared with those in S group and ES group, the serum levels of brain injury markers, apoptosis index (AI) and oxidative stress in hippocampus and cerebral cortex increased in I/R group and EI group (P < 0.05). Whereas, compared with those in I/R group, the above indicators in EI group decreased (P < 0.05). Therefore, hepatic I/R injury can lead to the damage of hippocampus and cerebral cortex, and the increased serum exosomes induced by hepatic I/R plays an important role.

The changes of intraoperative body temperature in adult liver transplantation: A retrospective study.

BACKGROUND: Body temperature is poorly regulated in patients with end-stage liver disease. Due to the prolonged surgery time and anhepatic time as well as the complex surgical procedures performed in liver transplantation, the body temperature fluctuates greatly. This study investigated the effect of intraoperative body temperature fluctuations on the prognosis of liver recipients. METHODS: The body temperatures of liver recipients recorded from the induction of anesthesia (T0) until the end of surgery (T14) were retrieved. The patients were divided into two groups: the hypothermia group (< 35 °C and ≥ 5 min) and the normothermia group (≥ 35 °C or < 35 °C but < 5 min). Intraoperative and postoperative variables were compared between the two groups, and the correlations between the duration of hypothermia and the medical variables were analyzed. RESULTS: Of the 107 patients, 67 patients were in the normothermia group, and 40 in the hypothermia group. The lowest body temperature was at 5 min after reperfusion for the whole cohort. Compared with the normothermia group, patients in the hypothermia group were more prone to bleeding, had a longer intubation time and increased rates of bacterial infection and acute pulmonary edema after liver transplantation (P < 0.05). Hypothermia time was positively correlated with bleeding volume, intubation time, units of blood transfusions and intensive care stay, but negatively correlated with urine output. CONCLUSIONS: The intraoperative body temperature exhibited a graphical "V" trend, and the lowest temperature was at 5 min after reperfusion. The longer the duration of hypothermia, the more unfavourable the prognosis.

Design of pH-sensitive peptides from natural antimicrobial peptides for enhancing polyethylenimine-mediated gene transfection.

BACKGROUND: Poor endosomal release is a major barrier of polyplex-mediated gene transfection. Antimicrobial peptides (AMPs) are commonly used to improve polyethylenimine (PEI)-mediated gene transfection by increasing endosomal release. In the present study, we designed novel pH-sensitive peptides that highly enhance transfection efficiency compared to their parent peptides. METHODS: Two analogues of melittin (Mel) and RV-23 (RV) were synthesized by replacing the positively-charged residues in their sequences with glutamic acid residues. The pH-sensitive lysis ability of the peptides, the effect of the peptides on physicochemical characteristics, the intracellular trafficking, the transfection efficiency, and the cytotoxicity of the polyplexes were determined. RESULTS: The acidic peptides showed pH-sensitive lytic activity. The hemolytic activity of acidic peptides at pH 5.0 was higher than that at pH 7.4. The incorporation of acidic peptides did not affect the DNA binding ability of PEI but affected the physicochemical characteristics of the PEI/DNA polyplexes, which may be beneficial for endosomal release and gene transfection. The incorporation of acidic peptides into PEI/DNA polyplexes enhanced the PEI-mediated transfection efficiency corresponding to up to 42-fold higher luciferase activity compared to that of PEI alone. CONCLUSIONS: The results of the present study indicate that replacement of positively-charged residues with glutamic acid residues in the AMP sequence yields pH-sensitive peptides, which enhance the transfection efficiency of PEI/DNA polyplexes in various cell lines.

Wuzi Yanzong pill, a Chinese polyherbal formula, alleviates testicular damage in mice induced by ionizing radiation.

BACKGROUND: Chinese medicine Wuzi Yanzong pill (WZYZP) was firstly documented in ancient Chinese medical works "She Sheng Zhong Miao Fang" by Shi-Che Zhang in 1550 AD. The traditional herbal formula is widely used in treating nephrasthenia lumbago, prospermia, erectile dysfunction and male sterility. The present study was to explore the effects of WZYZP on ionizing irradiation-induced testicular damage in mice. METHODS: The pelvic region of male mice was exposed to X-rays for inducing testicular damage. The effects of WZYZP on testicular damage were evaluated in terms of testes weight, sperm quantity and motility, testes oxidative status and serum hormone levels. The alterations in testicular structure were examined by hematoxylin-eosin staining. Additionally, changes in proliferating cell nuclear antigen (PCNA) expression of testes were explored by western blot. RESULTS: Pelvic exposure to x-ray induced reduction in testes weight and sperm quality, along with oxidative stress and abnormal testicular architecture in testes. Oral administration of WZYZP for 3 weeks markedly increased testes weight, sperm quantity and motility, and attenuated testicular architecture damage. Meanwhile, WZYZP treatment significantly reversed the reduction of serum testosterone, and decreased testes malondialdehyde (MDA) and Oxidative stress index (OSI) relative to the radiated mice. Additionally, WZYZP effectively prevented the downregulation of PCNA expression in testes induced by x-ray irradiation. CONCLUSION: These findings suggest WZYZP exhibits ameliorating effects against ionizing irradiation-induced testicular damage in mice, which may be related to its antioxidation.

Changes in central corneal thickness and refractive error after thin-flap laser in situ keratomileusis in Chinese eyes.

BACKGROUND: Refractive stability is influenced by alterations in corneal curvature and corneal thickness after laser in situ keratomileusis (LASIK). The aim of this study was to analyze the changes of central corneal thickness (CCT) and refractive error following thin-flap LASIK surgery in Chinese eyes. METHODS: One hundred and fifty-eight myopic patients (302 eyes) who underwent thin-flap LASIK surgery were prospectively evaluated. CCT was measured by non-contact specular microscopy before, and 1 day, 1 week, and 1, 3, and 6 months following surgery. Age, refractive error, and optic zone diameter were also recorded. RESULTS: Preoperatively, the mean CCT was 531.6 ± 24.3 µm. At 1 day, 1 week, and 1, 3, and 6 months after surgery, mean CCTs were 431.4 ± 38.4 µm, 422.6 ± 3 7.8 µm, 427.2 ± 38.0 µm, 434.4 ± 38.2 µm, and 435.6 ± 38.0 µm, respectively. Significant changes were detected in CCT values at each time point after thin-flap LASIK treatment (P < 0.05). The mean preoperative spherical equivalent (SE) was -5.73 ± 2.30 diopters (D). At 1 day, 1 week, and 1, 3, and 6 months after surgery, it was 0.26 ± 0.58 D, 0.54 ± 0.52 D, 0.49 ± 0.53 D, 0.45 ± 0.49 D, and 0.37 ± 0.42 D, respectively. The spherical equivalent refraction at 6 months postsurgery was close to the predicted value (0.34 ± 0.30 D). The changes in CCT within 6 months (4.06 ± 9.99 µm) were negatively correlated with age, preoperative refractive error, and optical zone diameter, respectively (r = -0.180, P < 0.05; r = -0.187, P < 0.001; r = -0.171, P < 0.05, respectively). No significant correlation was found between CCT changes and SE changes at different time points, postoperatively. CONCLUSIONS: CCTs decreased significantly at 1 day after surgery, and continued to decline at 1 week after surgery, then increased over time. From postoperative 1 week, SE over time continually shifted to the myopic side.

[Research on the characteristics of Ulva prolifera in Shandong Peninsula during 2008-2012 based on MODIS data].

In the present study, the MODIS data were used to monitor the situation of Ulva prolifera in the Shandong Peninsula waters during the period of 2008-2012. Those studies mainly calculate the area of NDVI, and get the information of the time, area , scope , floating path of Ulva prolifera by using threshold segmentation method. The feasibility of monitoring Ulva prolifera information based on MODIS data and the macroscopic regularity of the outburst of Ulva prolifera was elementally studied. The results showed that Ulva prolifera first generated in the middle of May or early June, the time, area, scope of Ulva prolifera reached a maximum, but the relative crowding density was earlier or later when Ulva prolifera developed into a outburst. Finally, Ulva prolifera died away after existing for 71 days in the late July or the early August. Wholly, the floating path moved to the northwest from off the coast to offshore. Based on those aspects above, the outburst of Ulva prolifera in 2008 and 2009 was more serious than others.

Glyphosate exposure affected longevity-related pathways and reduced survival in asian honey bees (Apis cerana).

Glyphosate (N-(phosphonomethyl)glycine, GLY) ranks among the most extensively used and effective herbicides globally. However, excessive GLY utilization poses a substantial threat to the survival of honey bees (Apis cerana). Here we monitored the survival status of A. cerana treated with GLY, and conducted transcriptome sequencing of the bee gut and head to further explore potential GLY influences at the molecular level. We observed that the mortality rate of bees increased as GLY concentration escalated. Pivotal pathways emerged in response to the GLY treatment, with a substantial number of differentially expressed genes enriched in the longevity regulating pathway - multiple species. This strongly suggested that GLY may influence the physiological behavior of bees by impacting this particular pathway. Moreover, our analysis revealed a notable reduction in the enzymatic activities of CYP450 and AChE in both the bee head and intestines of when exposed to GLY. Conversely, the enzymatic activity of superoxide dismutase (SOD) in the head remained unaffected, whereas in the intestines, it exhibited a significant increase. Additionally, prophenol oxidase (PPO) and glutathione-S-transferases (GSTs) displayed contrasting trends in enzymatic activity in both organs. This study offers valuable insights into how GLY impacted the survival of A. cerana.

Congenital absence of left atrial appendage in a patient with ischemic stroke and atrial fibrillation.

Left atrial appendage (LAA) is a finger-like muscular extension of the left atrium, and it is the most prominent site for cardiac thrombus in patients with atrial fibrillation. Congenital absence of LAA could be incidentally detected in patient with ischemic stroke and atrial fibrillation. Although it is considered to be an extremely rare cardiac anomaly, its clinical significance remains unknown and there is no clear consensus in the management strategy in those patients. Therefore, we report a case of an incidentally noted congenital absence of LAA in a 68-year-old woman being planned for LAA closure.

Bowman capsule rupture in children with myeloperoxidase-antineutrophil cytoplasmic antibody-associated glomerulonephritis predicts poor renal survival.

Background: Recent developments indicated that Bowman capsule rupture (BCR) is observed in antineutrophil cytoplasmic antibody (ANCA)-associated glomerulonephritis (AAGN). We aimed to explore the relationship between BCR and clinical manifestations, pathological changes, and prognosis in children with myeloperoxidase (MPO)-AAGN. Methods: A total of 56 children with MPO-AAGN were divided into BCR (+) and BCR (-) groups according to the status of Bowman's capsule. Clinical and histological features and renal outcomes were compared, and the predictive value of BCR for end-stage kidney disease (ESKD) of MPO-AAGN was evaluated. Results: After retrospective analysis of the data, 24 children (42.9%) were found to have BCR. The results showed that BCR positively correlated with intrarenal immune cell infiltrates, obsolescence and crescents in glomeruli, tubulointerstitial inflammation, tubulitis, and tubular atrophy negatively correlated with normal glomeruli and immunoglobulin G deposition in the kidney. The clinical features and kidney pathological changes were more severe in the BCR (+) group than BCR (-) group, and the renal survival rate was significantly poorer in the BCR (+) group than BCR (-) group (χ2 = 5.45, p = 0.02). Moreover, estimated glomerular filtration rate (≤15 mL/min/1.73 m2), BCR and ANCA renal risk score (ARRS) were independent risk factors for the development of ESKD in children with MPO-AAGN. After combining BCR with the Berden classification and ARRS, our data suggested that the Berden classification + BCR and ARRS + BCR showed better predictive values for ESKD than those of the Berden classification and ARRS, respectively. Conclusion: BCR is an important pathological lesion that correlates with severe clinical manifestations, pathological changes, and poor prognosis in children with MPO-AAGN.

[Detection and application of PIS genetic deficiency gene in dairy goat].

The purpose of this study was to develop a molecular method for detecting polled intersex syndrome (PIS) genetic deficiency gene in dairy goat. Three pairs of primers, PIS-, PIS+, and NEI were designed based on PIS gene sequence (AF404302) to identify the PIS genetic deficiency genotype. For the normal phenotype, the fragments of 141 and 300 bp were obtained for the genotype PIS-PIS-, and 141, 449, and 300 bp for the genotype PIS-PIS+. For the PIS goat with the genotype PIS+PIS+, 449 and 300 bp were obtained. Two hundred and twenty-four dairy goats in one population were tested based on this method. The results showed that there were 150 PIS-PIS+, 70 PIS -PIS-, and 4 PIS+PIS+. The genotype frequency of PIS-PIS+ was 66.9%, and the gene frequency of PIS+ was 35.3% in the population. Therefore, the frequency of PIS offspring was over 12%. This study developed a method to detect PIS genetic deficiency dairy goat. The method could identify buck genotype accurately to avoid the occurrence of PIS genetic deficiency. The ease and accuracy show a strong potential of the method for use in marker assisted selection of dairy goats and healthy development of dairy goat industry.

Effective treatment of high-voltage pulsed radiofrequency combined with oxygen-ozone injection in acute zoster neuralgia.

INTRODUCTION: Postherpetic neuralgia (PHN) is the most common and severe complication of acute herpes zoster. Early treatment of herpes zoster neuralgia is of great significance to reduce the incidence of PHN. This retrospective study evaluated the efficacy and safety of the combination of high-voltage pulsed radiofrequency (PRF) and oxygen-ozone(O2-O3) injection in patients with acute zoster neuralgia (AZN) who failed to respond to conservative treatment. METHODS: One-hundred patients diagnosed with AZN were classified into two groups (high-voltage PRF group [HP group, n = 50] and high-voltage PRF combined with O2-O3 injection group [HPO group, n = 50]) based on different treatment methods. Therapeutic effectiveness was assessed using a numerical rating scale (NRS) and the Pittsburgh Sleep Quality Index (PSQI). The dosages of gabapentin and tramadol (mg/d) before treatment and after 1 week and 1, 3, and 6 months of treatment were measured. The incidence of clinically meaningful PHN after treatment was also recorded. RESULTS: Pain intensity and sleep quality in both groups at all time points improved after treatment compared to before treatment (P < 0.05). After 1 week and 1 month of treatment, NRS and PSQI scores in both groups decreased, but the differences were not statistically significant (P > 0.05). NRS, PSQI scores, and the dosages of gabapentin and tramadol decreased more significantly in the HPO group than those in the HP group after 3 months (P < 0.05). The incidence of PHN was significantly lower in the HPO group than in the HP group (P < 0.05). There were no significant differences in adverse events between the groups. CONCLUSIONS: High-voltage PRF is a safe and effective method for treating AZN. The combination of high-voltage PRF and O2-O3 injection is superior to high-voltage PRF alone for treating late-stage AZN. This approach could be recommended as an alternative treatment for patients with refractory AZN and could significantly reduce the risk of PHN.

[Clinical observation of highly myopic eyes with retinoschisis after phacoemulsification].

OBJECTIVE: To evaluate the changes of the retinal thickness in the highly myopic eyes with retinoschisis after phacoemulsification by optical coherence tomography (OCT) and to analyze their postoperative visual acuity. METHODS: It was a retrospective case series study. This study included 25 (35 eyes) highly myopic patients with retinoschisis. One week, 1 and 3 months after phacoemulsification, the best corrected visual acuity (BCVA) was measured and OCT was performed to evaluate the central foveal thickness (CFT) as well as the maximum neurosensory thickness (MNT). These 35 eyes were divided into low and high accumulated energy complex parameter (AECP) groups, and ≤ 28.00 mm, > 28.00 and ≤ 30.00 mm, > 30.00 mm groups according to the axial length respectively. Variance analysis of data obtained from repeated measurement, Wilcoxon rank sum test, linear correlation and one-way ANOVA were used to analyze the data. RESULTS: According to the location of schisis, the 35 eyes were classified as foveal schisis (20 eyes) and nonfoveal schisis (15 eyes). Among the 35 eyes, there were 30 eyes with outer retinoschisis and 5 with compound schisis. The mean BCVA (logMAR) preoperatively and 1 week, 1 and 3 months postoperatively was 0.23 ± 0.24, 0.59 ± 0.38, 0.57 ± 0.38 and 0.60 ± 0.36 respectively (F = 31.15, P = 0.000). CFT preoperatively and 1 week, 1 and 3 months postoperatively was (255.46 ± 197.57) µm, (256.80 ± 199.88) µm, (274.37 ± 246.87) µm, and (268.60 ± 238.41) µm (F = 0.99, P = 0.420); while MNT was (473.31 ± 175.52) µm, (477.71 ± 188.71) µm, (486.60 ± 229.17) µm and (482.63 ± 208.82) µm respectively (F = 0.26, P = 0.857). BCVA in eyes with foveoschisis and in eyes with nonfoveal schisis was statistically significantly different at 1 week, 1 and 3 months postoperatively (Z = -0.580, P = 0.048; Z = -2.147, P = 0.030 and Z = -2.099, P = 0.034). BCVA (0.43 ± 0.28) was inversely correlated with CFT (497.42 ± 281.49) µm in the eyes with foveoschisis unaccompanied with a macular lamellar hole at 3 months postoperatively (r = -0.667, P = 0.018). There were no significant differences in the changes of the thickness of the retina between the groups with different AECP or among the groups with different axial lengths (Z = -0.314 - 1.290, P = 0.192−0.741 and F = 0.15−0.62, P = 0.545−0.859). CONCLUSIONS: Cataract surgery has no significant effect on thickness of the retina in the highly myopic eyes with retinoschisis. The patients with retinoschisis could achieve good visual recovery after phacoemulsification. Postoperative visual acuity is related to the location and degree of retinoschisis. Besides, patients with non-foveal schisis could obtain better visual acuity than those with foveoschisis. Further investigation on the development of retinoschisis and the complications after surgery is required.

A Machine Learning Study on the Thermostability Prediction of (R)-ω-Selective Amine Transaminase from Aspergillus terreus.

Artificial intelligence technologies such as machine learning have been applied to protein engineering, with unique advantages in protein structure, function prediction, catalytic activity, and other issues in recent years. Screening better mutants is still a bottleneck in protein engineering. In this paper, a new sequence-activity relationship method was analyzed for its application in improving the thermal stability of Aspergillus terreus (R)-ω-selective amine transaminase. The experimental data from 6 single-point mutated enzymes were used as a learning dataset to build models and predict the thermostability of 26 mutants. Based on digital signal processing (DSP), this method digitized the amino acid sequence of proteins by fast Fourier transform (FFT) and then established the best model applying partial least squares regression (PLSR) to screen out all possible mutants, especially those with high performance. In protein engineering, the innovative sequence activity relationship (ISAR) method can make a reasonable prediction using limited experimental data and significantly reduce the experimental cost. The half-life (T 1/2) of (R)-ω-transaminase was fitted with the amino acid sequence by the ISAR algorithm, resulting in an R 2 of 0.8929 and a cvRMSE of 4.89. At the same time, the mutants with higher T 1/2 than the existing ones were predicted, laying the groundwork for better (R)-ω-transaminase in the later stage. The ISAR algorithm is expected to provide a new technique for protein evolution and screening.

The Accumulation of Lutein and ß-Carotene and Transcript Profiling of Genes Related to Carotenoids Biosynthesis in Yellow Celery.

Carotenoids are the general term of natural pigments. The formation of plant color is probably related to the components of carotenoids. As the yellow variety of celery, it is rich in the composition and content of carotenoids. However, the transcript profiling and roles of the genes related to carotenoids biosynthesis in yellow celery remain unclear. In this study, three yellow celery cultivars at different growth stages were used to analyze the content and composition of carotenoids and transcriptional changes of carotenoid biosynthesis-related genes. The lutein and ß-carotene were detected in yellow celery cultivar, while α-carotene and lycopene were not detected. The contents of lutein and ß-carotene were higher in leaf blades than in petioles. During the growth and development, the contents of lutein and ß-carotene gradually decreased in celery. Compared with the other two cultivars, the contents of lutein and ß-carotene were the highest in 'Huangtaiji' of 65 days after sowing (DAS) and 85 DAS and 'Liuhehuangxinqin' of 105 DAS, respectively. The expression levels of AgLCYB and AgPSY2 genes were significantly correlated with lutein and ß-carotene contents. This work provided a reference for the further study on carotenoid metabolisms in yellow celery and also made sense on the way of cultivating yellow celery with high carotenoids content.

AgZDS, a gene encoding ζ-carotene desaturase, increases lutein and ß-carotene contents in transgenic Arabidopsis and celery.

ζ-Carotene desaturase (ZDS) is one of the key enzymes regulating carotenoids biosynthesis and accumulation. Celery transgenic efficiency is low and it is difficult to obtain transgenic plants. The study on ZDS was limited in celery. Here, the AgZDS gene was cloned from celery and overexpressed in Arabidopsis thaliana and celery to verify its function. The AgZDS has typical characteristic of ZDS protein and is highly conserved in higher plants. Phylogenetic analysis showed that AgZDS has the closest evolutionary relationship with ZDSs from Solanum lycopersicum, Capsicum annuum and Tagetes erecta. Overexpression of AgZDS gene in A. thaliana and celery resulted in increased accumulations of lutein and ß-carotene and up-regulated the expression levels of the genes involved in carotenoids biosynthesis. The contents of lutein and ß-carotene in two lines, AtL1 and AgL5, were the highest in transgenic A. thaliana and celery, respectively. The relative expression levels of 5 genes (AtPDS, AtZISO, AtZEP, AtNCED3, and AtCCD4) were up-regulated compared to the wild type plants. The relative expression levels of most genes in carotenoids biosynthesis pathway, such as AgPDS, AgCRTISO1, and AgZISO, were up-regulated in transgenic celery plants. The antioxidant capacity of A. thaliana and photosynthetic capacity of celery were also enhanced. This research is the first report on the function of structure gene related to carotenoid biosynthesis in transgenic celery plants. The findings in this study demonstrated the roles of AgZDS in regulating carotenoids metabolism of celery, which laid a potential foundation for quality improvement of celery.

[Cerebral lymphatic blockage aggravates apoptosis of cortical neurons after subarachnoid hemorrhage in rats in vivo].

This present study was performed to investigate the influence of cerebral lymphatic blockage (CLB) on apoptosis of cortical neurons after subarachnoid hemorrhage (SAH) in rats in vivo. Healthy adult Wistar rats were randomly divided into normal control group, SAH group and SAH+CLB group. SAH model was made by double injection of autologous blood into the cisterna magna. On the third day after the second cisternal injection, morphological changes of cortical cells were observed by hematoxylin-eosin (HE) combined with propidium iodide (PI) staining. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method was applied to determine in situ apoptosis in the cerebral cortex. Immunohistochemistry was conducted to detect the expression of Caspase-3 and Bcl-2 in cortical neurons. HE and PI staining showed that cortical neurons of SAH rats were partly shrinkable; the nuclei showed wavy, folded or wrinkled appearance, and some nuclei had the shape of crescent. The cortical neurons in SAH+CLB group distributed sparsely and the nuclear fragmentation, apoptotic bodies were found, surrounded by the formation of vacuoles. The numbers of TUNEL-positive cells in SAH group and SAH+CLB group were higher than that in the normal control group, while the number in SAH+CLB group was significantly higher than that in the SAH group. Caspase-3 expressions in SAH group and SAH+CLB group were higher than that in the normal control group, while the expression in SAH+CLB group was significantly higher than that in the SAH group. Bcl-2 expressions in SAH group and SAH+CLB group were higher than that in the normal control group, while the expression in the SAH+CLB group was significantly lower than that in SAH group. The results obtained suggest that CLB exacerbates the apoptosis of cortical neurons in rats after SAH by up-regulating Caspase-3 expression and down-regulating Bcl-2 expression.

An alternative pathway for cellular protection in BRAF inhibitor resistance in aggressive melanoma type skin cancer.

Oncogenic alterations in the BRAF gene are identified in an estimate of 50% of melanomas and cause melanoma development. BRAF kinase inhibitors (BRAFi), including vemurafenib and dabrafenib, were discovered and used in the clinical treatment of BRAF-mutant metastatic melanoma. Though, BRAFi's therapeutic advantages are short term and short-lived associated with drug resistance. Although a few pathways of developed BRAFi resistance have also been established, in approximately 40% of melanomas, the cause for inherited resistance remains unclear. Recognizing a new process of developed BRAFi resistance might provide new possibilities to successfully treat BRAF mutant melanoma. In this study, we are exploring the compensatory alternative pathway followed by BRAFi/MEKi treated resistant cell for maintaining the long-term integrity and survival.

Protective effects of green tea extracts on photoaging and photommunosuppression.

OBJECTIVES: This study aimed to investigate whether the sunscreen-containing 2-5% green tea extracts (GTEs) protect ultraviolet irradiation (UVR)-induced photoaging and photoimmunosuppression. MATERIALS AND METHODS: Twenty volunteers were exposed to repetitive solar-simulated UVR (ssUVR) on the upper back at a dosage of 1.5 minimal erythema doses (MED) per day for four consecutive days. Thirty minutes before each UVR and 6, 24, and 48 h after the last UV exposure, the products containing vehicle, and 2-5% GTEs were applied onto five sites on the dorsal skin, respectively. The skin biopsies were obtained 72 h after the last UVR. The thickness of the stratum corneum and epidermis was measured under the microscope and the expression of cytokeratins (CK)-5/6, CK16, metalloproteinases (MMP)-2, MMP-9, and the CD1a(+) Langerhans cells (LCs) were determined using immunohistochemistry. RESULTS: Our results showed that UVR substantially induced cutaneous erythema, thickening of the epidermis, overexpression of CK5/6, CK16, MMP-2, MMP-9, and depletion of CD1a(+) LCs. The sunscreens containing different concentrations of GTEs conferred significant protection against the photoaging and photoimmunology-related biological events. Interestingly, the protective effects were not parallel to the concentrations of GTEs, with 2% and 3% GTEs showing the most efficacious photoprotection. CONCLUSIONS: GTEs-containing sunscreens have potential photoprotective effects on UVR-induced photoaging and photoimmunosuppression.