Influential Factors in the Efficacy of Hemoporfin-Mediated Photodynamic Therapy for Port-wine Stains.

Hemoporfin-mediated photodynamic therapy (HMME-PDT) is commonly used in the treatment of port-wine stains (PWS). However, the influential factors for the efficacy of the treatment are not well defined. This study intends to observe the influential factors for the efficacy of HMME-PDT in the treatment of port-wine stains (PWS). A total of 551 patients with PWS of head and neck was enrolled in this retrospective study. Further screening the patients of facial PWS, 484 patients were chosen. Patients were treated with HMME-PDT. All patients received 1~3 sessions of treatment with 2~3-month intervals. We photographed the lesions before each session and 2~3 months after the last session. Ages, sessions, lesion subtypes, and previous treatment history were related to the response of HMME-PDT (P =0.032, P<0.001, P=0.012, P=0.003 respectively). Treatment sessions were the independent factor correlated with efficacy after 3 sessions of treatment. Patients with no treatment history targeting PWS showed higher efficacy than those were treated with laser or other photodynamic treatment (P<0.05). The efficacy was higher by increasing the sessions of treatment. The efficacy was higher for lesion on maxillary prominence area and mandibular prominence area that on frontonasal prominence area and optic vesicle area (P<0.05). HMME-PDT is an effective in the treatment of PWS. Patients received no previous treatment for PWS, total treatment sessions and lesion on maxillary prominence area and mandibular prominence area are positive factors.

Targeting critical pathways in ferroptosis and enhancing antitumor therapy of Platinum drugs for colorectal cancer.

Colorectal cancer (CRC) can be resistant to platinum drugs, possibly through ferroptosis suppression, albeit the need for further work to completely understand this mechanism. This work aimed to sum up current findings pertaining to oxaliplatin resistance (OR) or resistance to ascertain the potential of ferroptosis to regulate oxaliplatin effects. In this review, tumor development relating to iron homeostasis, which includes levels of iron that ascertain cells' sensitivity to ferroptosis, oxidative stress, or lipid peroxidation in colorectal tumor cells that are connected with ferroptosis initiation, especially the role of c-Myc/NRF2 signaling in regulating iron homeostasis, coupled with NRF2/GPX4-mediated ferroptosis are discussed. Importantly, ferroptosis plays a key role in OR and ferroptotic induction may substantially reverse OR in CRC cells, which in turn could inhibit the imbalance of intracellular redox induced by oxaliplatin and ferroptosis, as well as cause chemotherapeutic resistance in CRC. Furthermore, fundamental research of small molecules, ferroptosis inducers, GPX4 inhibitors, or natural products for OR coupled with their clinical applications in CRC have also been summarized. Also, potential molecular targets and mechanisms of small molecules or drugs are discussed as well. Suggestively, OR of CRC cells could significantly be reversed by ferroptosis induction, wherein this result is discussed in the current review. Prospectively, the existing literature discussed in this review will provide a solid foundation for scientists to research the potential use of combined anticancer drugs which can overcome OR via targeting various mechanisms of ferroptosis. Especially, promising therapeutic strategies, challenges ,and opportunities for CRC therapy will be discussed.

Effectiveness of Trimetazidine in Patients with Chronic Heart Failure Stratified by the Expression of Soluble Suppression of Tumorigenicity-2 (sST2): A Prospective Cohort Study.

INTRODUCTION: Trimetazidine has been reported to have potential benefits in patients with chronic heart failure (CHF). Soluble suppression of tumorigenicity-2 (sST2) was shown to worsen CHF and, hence, has a diagnostic value in heart failure. The aim of the present study was to evaluate the effectiveness of trimetazidine in patients expressing high and low levels of sST2 compared with their matched placebo. METHODS: In this prospective cohort study, 170 patients were enrolled. Patients expressing more than 35 ng/mL sST2 (S+) were split into a trimetazidine group (group A) and placebo group (group B). Likewise, patients expressing 35 ng/mL or less of sST2 (S-) were divided into a trimetazidine (group C) and placebo group (group D). Patients in both the trimetazidine groups were administered 20-mg twice-a-day doses of trimetazidine. Trimetazidine effectiveness was determined in terms of changes in cardiac function, motor function, and mental status at 1, 3, 6, and 12 months from baseline among the four groups. RESULTS: A total of 158 patients were included for final data analysis (group A, n = 50; group B, n = 57; group C, n = 27; group D, n = 24). On comparing different outcomes between the four groups and across the time points, significant difference was observed between the groups in ejection fraction (EF; P < 0.001), cardiac index (CI; P < 0.001), New York Heart Association score (P < 0.001), 6-min walk test (P < 0.001), Veterans Specific Activity Questionnaire (VSAQ; P < 0.001), Minnesota Living with Heart Failure Questionnaire (MLHFQ; P < 0.001), hospital anxiety and depression scores (P < 0.001), and Copenhagen Burnout Inventory (P < 0.001). Significant difference in systolic blood pressure (P < 0.001), heart rate (P < 0.001), EF (P < 0.001), CI (P < 0.001), VSAQ (P = 0.017), and MLHFQ (P < 0.001) was observed. CONCLUSION: Trimetazidine demonstrated an overall improvement in cardiac function, motor function, quality of life (QoL), and mental status in both S+ and S- patients. Among patients administered trimetazidine, significant changes in maximum outcomes were observed among those expressing higher levels of sST2 compared with placebo.

Solid matrix priming improves cauliflower and broccoli seed germination and early growth under the suboptimal temperature conditions.

Seed priming is an effective method for imparting stress tolerance to plants. This study aimed to analyze the effects of solid matrix priming (SMP) on cauliflower and broccoli seed germination and early seedling growth under suboptimal temperature conditions. The SMP method used in this study included the following steps: (1) mixing seeds with vermiculite and water at a ratio of 2:3:2.5 (w/w/v) and incubating for 2 days in the dark at 20°C; (2) drying the SM-primed seed; (3) germinating the SM-primed and the nonprimed seeds at 10, 15, 20, and 25°C; (4) analyzing the antioxidant enzyme activities of SM-primed and nonprimed germinating broccoli and cauliflower seeds in the early germination stage at 10, 15, 20, and 25°C; and (5) testing the emergence of SM-primed and nonprimed control seeds in the early spring glasshouse. The results showed that the SMP improved seed germination vigor and early seedling growth and increased the activities of peroxidase and ascorbate peroxidase in the germinating cauliflower and broccoli seeds under the suboptimal temperature conditions in the early germination stage compared with nonprimed seeds. It was observed that the suboptimal temperature conditions (i.e., 10 and 15°C) suppressed SM-primed and nonprimed seed germination and early seedling growth of cauliflower and broccoli. Inside a greenhouse, the SMP improved the emergence of cauliflower and broccoli seeds during the early spring season. SMP is an effective method for improving seed germination and the emergence of cauliflower and broccoli under suboptimal temperature conditions.

A case of a death caused by an atrial-oesophageal-thoracic fistula after radiofrequency ablation of atrial fibrillation.

The report is about a 49-year-old man with rheumatic heart disease and atrial fibrillation. He underwent mitral valve replacement, tricuspid valvuloplasty, and atrial fibrillation radiofrequency ablation in the hospital. He vomited blood on the 2nd postoperative day, and the bleeding gradually worsened thereafter. He had to have repeated drainage of large amounts of blood from his right thoracic cavity and digestive tract. He died suddenly after undergoing an oesophageal endoscopy on the 24th postoperative day. The autopsy revealed an atrial-oesophageal-thoracic fistula. By excluding the possibility of the fistula being caused by complications from nasoenteric feeding, tracheal intubation, and a foreign body ingestion, we determined that the atrial-oesophageal-thoracic fistula was a complication after radiofrequency ablation according to the finding of coagulation necrosis of the myocardial cells at the left atrium fistula. In addition, we also performed an elemental analysis on the radiofrequency ablation area and other cardiac tissues by scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS) and found five metal elements, Cr, Cu, Zn, Mn, and Ti, which specifically existed in the radiofrequency ablation area. This finding has the potential to serve as new evidence for radiofrequency ablation and is a worthy direction of research.

Simultaneous determination of 16 important biologically active phytohormones in Dendrobium huoshanese by pressurized capillary electrochromatography.

A rapid pressurized capillary electrochromatography (pCEC) method has been successfully developed for the simultaneous determination of 16 phytohormones in Dendrobium huoshanense. Effects of wavelength, mobile phase, the flow rate, pH value, concentration of buffer and applied voltage were investigated, respectively. The results showed that the 16 phytohormones could be baseline-separated rapidly in less than 21 min on a reversed-phase EP-100-20/45-3-C18 capillary column (total length of 45 cm, effective length of 20 cm, diameter of 100 µm, ODS packing inside for 3 µm) with ACN/5.0 mM ammonium acetate (containing 0.05% formic acid, pH = 3) as the mobile phase using gradient elution mode as follows: 0.1-10.0 min 40%ACN,10-15.0 min 70%ACN, 15.0-20 min 80% ACN, 20-21.0 min 80% ACN at a flow rate of 0.12 mL/min, applied voltage of -5 kV and a UV detection wavelength of 210 nm. The method validation howed that the established method is precise and stability, and the RSDs of intra- and inter-day precision based retention time and peak area were all below 5%. Employed the established method, in our experimental conditions, total 6 endogenous hormones including IAA, IBA, NAA, GA, ABA, t-Z were detected in D. huoshense. However, a relative larger amount of exogenous hormone 2,4-D (25.3 ~ 4.2 µg/kg) and 6-BA (79.5 ~ 35.4 µg/kg) were detected in 1 ~ 4 year old cultivated D. huoshense, suggesting there were still a certain amount of exogenous hormone residue in tissue-cultured D. huoshanese though they had been transplanted to field cultivation from the test-tube plantlets for several years.

IFI27 as a potential indicator for severe Enterovirus 71-infected hand foot and mouth disease.

The pathogenesis of Enterovirus 71 (EV71)-induced severe hand foot and mouth disease (HFMD) has not yet been clearly established. Further study into the mechanisms underlying host immune responses to EV71 infection and identifying important predictors will be crucial to antiviral treatment and early recognition of severe HFMD. The present study establishes that T help (Th)1 type, Th2 type, and Th17 type cytokine levels in serum of peripheral blood from patients with severe HFMD is higher than in peripheral blood from healthy subjects. The most significant increase occurred as the IL-6. In order to identify the important molecules in peripheral blood mononuclear cells (PBMCs) from severe HFMD patients, we performed transcriptome sequencing analysis of PBMC from severe HFMD patients and compared them to healthy controls. Interferon α-inducible protein 27 (IFI27) and cluster of differentiation 27 (CD27) were found to be the most significant differentially expressed gene. Finally, IFI27 was proved to be present at higher levels in patients with severe HFMD than in patients with mild HFMD. Our results suggest that IFI27 may be an indicator of the severity of cases EV71-induced HFMD.

Effects of high-intensity interval training on adipose tissue lipolysis, inflammation, and metabolomics in aged rats.

High-intensity interval training (HIIT) is a time-efficient alternative to moderate-intensity continuous training (MICT) to improve metabolic health in older individuals. However, differences in adipose tissue metabolism between these two approaches are unclear. Here, we evaluated the effects of HIIT and MICT on metabolic phenotypes in aged rats. HIIT significantly decreased fat mass, increased percent lean mass, decreased fat-to-lean ratio, reduced serum high-sensitivity C-reactive protein, increased serum interleukin-10 levels, and decreased perirenal adipose tissue leptin compared with rats in the sedentary (SED) group. HIIT also increased pregnenolone, cortisol, and corticosterone in both adipose tissue and serum. Both exercise modalities enhanced hormone-sensitive lipase and adipose triglyceride lipase expression compared with the SED group and decreased palmitic acid, stearic acid, octadecadienoic acid, urea, 1-heptadecanol, and α-tocopherol. MICT was related to glycerolipid metabolism, whereas HIIT was related to steroid hormone biosynthesis. Overall, HIIT showed more favorable regulation of anti-inflammatory activity than MICT.

Proteomics-based identification of different training adaptations of aged skeletal muscle following long-term high-intensity interval and moderate-intensity continuous training in aged rats.

Aging-associated loss of skeletal muscle mass and force increases the risk of falls, impairs mobility, and leads to a reduced quality of life. High-intensity interval training (HIIT) is superior to moderate-intensity continuous training (MICT) for improving morphological and metabolic adaptations of skeletal muscle in older adults, but the underlying mechanism is unknown. Aged female rats underwent HIIT and MICT for 8 months, and their differential impacts on skeletal muscle proteome were investigated. HIIT resulted in a larger improvement in grip strength and fiber cross-sectional area, with similar increases in inclined plane performance and time to exhaustion. Proteomic analysis showed that common training adaptations of both protocols included changes to muscle contraction, focal adhesion signaling, mitochondrial function, apoptosis and regeneration, and anti-oxidation, whereas protein processing in the endoplasmic reticulum and adipocytokine signaling were specifically altered in the MICT and HIIT groups, respectively. Immunoblotting showed that upregulation of the adiponectin/AMPK signaling pathway may be associated with improvements in autophagy, oxidative stress, mitochondrial function, and apoptosis in aged skeletal muscle following HIIT. Thus, understanding the molecular differences in training adaptations from these two exercise modalities may aid in combatting sarcopenia.

Beneficial Autophagic Activities, Mitochondrial Function, and Metabolic Phenotype Adaptations Promoted by High-Intensity Interval Training in a Rat Model.

The effects of high-intensity interval (HIIT) and moderate-intensity continuous training (MICT) on basal autophagy and mitochondrial function in cardiac and skeletal muscle and plasma metabolic phenotypes have not been clearly characterized. Here, we investigated how 10-weeks HIIT and MICT differentially modify basal autophagy and mitochondrial markers in cardiac and skeletal muscle and conducted an untargeted metabolomics study with proton nuclear magnetic resonance (1H NMR) spectroscopy and multivariate statistical analysis of plasma metabolic phenotypes. Male Sprague-Dawley rats were separated into three groups: sedentary control (SED), MICT, and HIIT. Rats underwent evaluation of exercise performance, including exercise tolerance and grip strength, and blood lactate levels were measured immediately after an incremental exercise test. Plasma samples were analyzed by 1H NMR. The expression of autophagy and mitochondrial markers and autophagic flux (LC3II/LC3-I ratio) in cardiac, rectus femoris, and soleus muscle were analyzed by western blotting. Time to exhaustion and grip strength increased significantly following HIIT compared with that in both SED and MICT groups. Compared with those in the SED group, blood lactate level, and the expression of SDH, COX-IV, and SIRT3 significantly increased in rectus femoris and soleus muscle of both HIIT and MICT groups. Meanwhile, SDH and COX-IV content of cardiac muscle and COX-IV and SIRT3 content of rectus femoris and soleus muscle increased significantly following HIIT compared with that following MICT. The expression of LC3-II, ATG-3, and Beclin-1 and LC3II/LC3-I ratio were significantly increased only in soleus and cardiac muscle following HIIT. These data indicate that HIIT was more effective for improving physical performance and facilitating cardiac and skeletal muscle adaptations that increase mitochondrial function and basal autophagic activities. Moreover, 1H NMR spectroscopy and multivariate statistical analysis identified 11 metabolites in plasma, among which fine significantly and similarly changed after both HIIT and MICT, while BCAAs isoleucine, leucine, and valine and glutamine were changed only after HIIT. Together, these data indicate distinct differences in specific metabolites and autophagy and mitochondrial markers following HIIT vs. MICT and highlight the value of metabolomic analysis in providing more detailed insight into the metabolic adaptations to exercise training.

Research progress in machine learning methods for gene-gene interaction detection.

Complex diseases are results of gene-gene and gene-environment interactions. However, the detection of high-dimensional gene-gene interactions is computationally challenging. In the last two decades, machine-learning approaches have been developed to detect gene-gene interactions with some successes. In this review, we summarize the progress in research on machine learning methods, as applied to gene-gene interaction detection. It systematically examines the principles and limitations of the current machine learning methods used in genome wide association studies (GWAS) to detect gene-gene interactions, such as neural networks (NN), random forest (RF), support vector machines (SVM) and multifactor dimensionality reduction (MDR), and provides some insights on the future research directions in the field.

The clinical significance and underlying correlation of pStat-3 and integrin αvß6 expression in gallbladder cancer.

BACKGROUND: Both phosphorylated signal transducer and activator of transcription 3(pStat-3) and integrin αvß6 can play vital role in the development and progression of cancer. However, little is known about their expression correlation and clinical significance in gallbladder cancer(GBC). OBJECTIVE: The aim of our present study was to investigate the expression of pStat-3 and integrin αvß6, two proteins' correlation and their clinical significance in GBC tissues. RESULTS: The expression of pStat-3 and integrin αvß6 were both significantly associated with T stage, lymph node metastasis status, TNM stage (P=0.008, P=0.000, P=0.000 and P=0.036, P=0.001,P=0.000,respectively). IHC and Western blot showed their expressions in GBC tissues were higher than that in paraneoplastic tissues. Moderate positive correlation existed between the two proteins (r =0.349, P <0.001). The survival analysis by Kaplan-Meier and Cox regression model showed that GBC patients with pStat-3 or integrin αvß6 positive expression had a significantly poorer 2-year survival rate (P = 0.002 and 0.000, the log-rank test, respectively), and either marker could act as unfavorable independent prognostic factors(RR=1.907, P=0.021 and RR=2.046, P=0.038). MATERIALS AND METHODS: The expression levels of pStat-3 and integrin αvß6 were analyzed in GBC cancerous and paraneoplastic tissues of 97 cases via immunohistochemistry(IHC) and further validated by western blot method. Besides, SPSS software was used to observe their clinical significance as well as the two proteins' correlation. CONCLUSION: pStat-3 and integrin αvß6 were indicators of tumor's progression and poor prognosis of patients with GBC. And the further study involving them may provide a helpful therapeutic target in prevention and treatment of GBC patients.

Serum atrial natriuretic peptide (ANP) as an objective indicator for the diagnosis of neurogenic shock: animal experiment and human case report.

In forensic medicine, the diagnosis of death due to neurogenic shock is considered to be an aporia, as lacking objective indicators and presenting atypical symptoms in autopsy. Medico-legal disputes and complaints occasionally result from this ambiguity. To explore potential objective indicators of neurogenic shock, we set up a model of neurogenic shock by applying an external mechanical force on the carotid sinus baroreceptor in rabbits. The serum atrial natriuretic peptide (ANP) level was measured by radioimmunoassay in the control group (n = 8), survival group (n = 15) and death group (n = 5) both before and after the insult. The serum ANP level showed a significant increase after the insult in the death group compared with the serum obtained before the insult (P = 0.006), while the serum ANP level after the insult in the survival group and control group was not statistically significant compared with the serum obtained before the insult (P = 0.332 and P = 0.492, respectively). To verify the repeatability of the model and the postmortem behavior of serum ANP, five healthy adult rabbits underwent the same procedure as the experimental group. The mortality rate was consistent with the former experiment (20 %). There were no significant changes in serum ANP level in vitro and in vivo (within 48 and 24 h, respectively). But there was a significant decrease in serum ANP level at 48 h postmortem in vivo (P = 0.001). A female patient who expired due to neurogenic shock during a hysteroscopy was reported. Neither fatal primary disease nor evidence for mechanical injuries or intoxication was found according to the autopsy. The serum ANP level was assayed as a supplementary indicator and was found to be three-fold higher than the normal maximum limit. Combined with the animal experiment, this case highlights that serum ANP has the potential to be an objective indicator for the diagnosis of death due to neurogenic shock.

[Comparative analysis of malaria detection ability of laboratories in Shanghai City from 2012 to 2015].

OBJECTIVE: To compare the application effects of three methods, namely microscopic examination, antigen detection (RDT) and nucleic acid test (PCR) in malaria detection between municipal and districts/counties centers for disease control and prevention in Shanghai, and analyze the malaria detection ability of the laboratories in Shanghai. METHODS: The blood smears, whole blood samples, case review confirmation records and case data of malaria cases and suspected cases in Shanghai from 2012 to 2015 were collected by Shanghai Municipal Center for Disease Control and Prevention, and the detection results were analyzed and compared. RESULTS: A total of 212 samples with complete data were submitted by all districts (counties) in Shanghai from 2012 to 2015, the samples submitted by Jinshan Districts were the most (41.98%), and among the first diagnosis hospitals, those submitted by the tertiary hospitals were the most (82.07%). The submitted samples in the whole year were increased gradually from January to October. All the 212 samples were detected by three methods (the microscopic examination, RDT and PCR) in the laboratory of Shanghai Municipal Center for Disease Control and Prevention, and 167 were tested and confirmed comprehensively as positives, accounting for 78.77%, and 45 were confirmed as negatives, accounting for 21.23%. The samples were detected by the method of microscopy and domestic RDT in the laboratories of the centers for disease control and prevention at district/county level, totally 153 were tested as positives, accounting for 72.17%, 41 were unclassified, accounting for 19.34%, 53 were negative, accounting for 25.00%, and 6 were undetected, accounting for 2.83%. The coincidence of microscopic examination between the report hospitals and the centers for disease control and prevention at district/county level was 78.16%, and the coincidence between centers for disease control and prevention at district/county level and municipal level was 93.20%. The utilization rate of RDT in the laboratory of district/county level was 73.58%. The coincidence of RDT tests between those domestic and imported was 93.59%. Compared with the detection results by municipal center for disease control and prevention, 37 samples were misjudged by the laboratories of district/county level. Almost all (99.37%) of the confirmed malaria cases were imported overseas, including Africa (85.44%), Asia (13.92%) and America (0.63%). CONCLUSIONS: The surveillance after malaria elimination in Shanghai should be carried out by combining with different detection methods and resource integration.

Oxidored-nitro domain-containing protein 1 expression is associated with the progression of hepatocellular carcinoma.

Hepatocarcinogenesis is a stepwise process during which multiple genes are altered. Understanding the molecular mechanisms that induce hepatocarcinogenesis may improve the screening, prevention and treatment of patients with hepatocellular carcinoma (HCC). In recent years, the oxidored-nitro domain-containing protein 1 (NOR1) gene has been identified to have an important role in the development of HCC in vitro experiments. The current study aimed to examine the expression of NOR1 mRNA and protein expression in specimens of normal liver, hepatitis, cirrhosis and HCC, together representing the process of HCC development. Furthermore, the association between NOR1 expression and clinicopathological parameters of HCC patients was analyzed. Tissue microarrays containing the specimens of human normal liver, hepatitis, cirrhosis and HCC were purchased, and in situ hybridization and immunohistochemistry were used to detect the expression of NOR1 mRNA and protein expression, respectively. It was revealed that the positive rate of NOR1 protein and mRNA expression in the specimens of hepatitis and cirrhosis were not significantly different from that in the normal liver samples. However, the specimens of HCC exhibited an increased positive rate of NOR1 protein and mRNA expression in comparison with the normal liver samples. In addition, a higher positive rate of NOR1 protein expression was observed in HCC patients with a poor pathological differentiation grade and high tumor node metastasis (TNM) stage. In conclusion, the present study provides evidence, for the first time, of the increased expression of NOR1 in human HCC tissues, and its correlation with the pathological stage and TNM status. These findings indicate that NOR1 may be involved in the progression of HCC and it could be employed as a predictive biomarker in HCC development.

[Investigation of two blood parasitic protozoa infection in farmed Macaca fascicularis in Guangxi Zhuang Autonomous Region].

OBJECTIVE: To investigate the infection situation of blood parasitic protozoa in farmed Macaca fascicularis in an animal breeding ground in Nanning, Guangxi Zhuang Autonomous Region, so as to provide the evidence for the prevention and control of human blood parasitic protozoa. METHODS: A total of 993 blood samples from farmed M. fascicularis were collected and stored on FTA cards. Among them, 550 thin blood smears were made. Each 10 samples were mixed in groups, and then the Babesia spp. and Plasmodium spp. in the blood of M. fascicularis were detected by Nest-PCR and PCR, respectively. The positive groups were tested individually. The thin blood smears stained with Giemsa were examined microscopically when PCR reported the samples were positive. RESULTS: When detected by Nest-PCR, the positive rate of Babesia. microti was 6.95% (69/993); only 1 positive sample with Plasmodium inui was detected by PCR. Among the 22 positive thin blood smears detected by PCR, 16 were determined with B. microti by microscopic examinations, on which the ring forms could be observed in the erythrocytes, but no hemozoin. CONCLUSIONS: The positive rate of B. microti in M. fascicularis in Guangxi Zhuang Autonomous Region is high, and the animal may play a role as a reservoir host in the transmission of B. microti. In the screening of B. microti with low infection density, Nest-PCR has a higher sensitivity.

[Cloning and characterization of ß-carbonic anhydrase, a potential drug target of Schistosoma japonicum].

OBJECTIVE: To express the beta carbonic anhydrase (ß-CA) of Schistosoma japonicum, and analyze its catalytic activity. METHODS: The cDNA and amino sequence which may encode ß-CA of S. japonicum were obtained by the bioinformatics-method, and then the cDNA sequence was cloned into prokaryotic expression vector pET-32a (+) and expressed. After examining by SDS-PAGE and Western blotting, the recombinant protein was purified by Ni-affinity chromatography and the catalytic activity was determined. RESULTS: The sequence Sjp_0056790.1 took on the conservative position of ß-CAs. The PCR and restriction enzyme digestion confirmed the construction of recombinant plasmid pET-32a (+) -SjaCA. SDS-PAGE and Western blotting analyses showed that the molecular weight of recombinant protein was about 38 kDa as expected, and it could be recognized by anti-His tag antibody. The catalytic activity determining revealed that the recombinant protein SjaCA owned the carbonic anhydrase activity. CONCLUSIONS: Sjp_0056790.1 encodes the ß-CA of S. japonicum, and the ß-CA with catalytic activity is successfully expressed, so it lays a foundation for the subsequent research of pharmacological inhibition, providing theoretic basis for searching and developing a new feasible anti-schistosome drug.

A salivary sheath protein essential for the interaction of the brown planthopper with rice plants.

Salivary secretions, including gel saliva and watery saliva, play crucial roles in the interaction between the insect and plant during feeding. In this study, we identified a salivary gland-specific gene encoding a salivary sheath protein (NlShp) in Nilaparvata lugens. NlShp has two alternative splicing variants; both are expressed at high levels during the nymph and adult stages. Immunohistochemical staining showed that the NlShp were synthesized in the principal gland cells of the salivary gland. LC-MS/MS and western blot analysis confirmed that NlShp was one of the components of the salivary sheath. Simultaneously knocking down the two NlShp variants by RNA interference inhibited both salivary flange and salivary sheath formation and resulted in a lethal phenotype within four days for the brown planthopper (BPH) feeding on rice plants, indicating that the salivary sheath and salivary flanges were essential for plant-associated feeding. Despite the salivary sheath deficiency, no obvious phenotype was observed in the NlShp-knockdown BPHs fed on artificial diet. The electrical penetration graph (EPG) results showed that salivary sheath-deficient BPHs exhibited a prolonged nonpenetration period, scarce sap period, and increased stylet movement on rice plants and eventually starved to death. Our results provided evidence that the interaction between the salivary sheath and host plant might be a critical step in successful BPH feeding. According to present research, we propose a salivary sheath required feeding model for piercing-sucking insects and provide a potential target for rice planthopper management.

Nucleosome alterations caused by mutations at modifiable histone residues in Saccharomyces cerevisiae.

Nucleosome organization exhibits dynamic properties depending on the cell state and environment. Histone proteins, fundamental components of nucleosomes, are subject to chemical modifications on particular residues. We examined the effect of substituting modifiable residues of four core histones with the non-modifiable residue alanine on nucleosome dynamics. We mapped the genome-wide nucleosomes in 22 histone mutants of Saccharomyces cerevisiae and compared the nucleosome alterations relative to the wild-type strain. Our results indicated that different types of histone mutation resulted in different phenotypes and a distinct reorganization of nucleosomes. Nucleosome occupancy was altered at telomeres, but not at centromeres. The first nucleosomes upstream (-1) and downstream (+1) of the transcription start site (TSS) were more dynamic than other nucleosomes. Mutations in histones affected the nucleosome array downstream of the TSS. Highly expressed genes, such as ribosome genes and genes involved in glycolysis, showed increased nucleosome occupancy in many types of histone mutant. In particular, the H3K56A mutant exhibited a high percentage of dynamic genomic regions, decreased nucleosome occupancy at telomeres, increased occupancy at the +1 and -1 nucleosomes, and a slow growth phenotype under stress conditions. Our findings provide insight into the influence of histone mutations on nucleosome dynamics.