Involvement of Three Esterase Genes from Panonychus citri (McGregor) in Fenpropathrin Resistance.

The citrus red mite, Panonychus citri (McGregor), is a major citrus pest with a worldwide distribution and an extensive record of pesticide resistance. However, the underlying molecular mechanism associated with fenpropathrin resistance in this species have not yet been reported. In this study, synergist triphenyl phosphate (TPP) dramatically increased the toxicity of fenpropathrin, suggesting involvement of carboxylesterases (CarEs) in the metabolic detoxification of this insecticide. The subsequent spatiotemporal expression pattern analysis of PcE1, PcE7 and PcE9 showed that three CarEs genes were all over-expressed after insecticide exposure and higher transcripts levels were observed in different field resistant strains of P. citri. Heterologous expression combined with 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetra-zolium bromide (MTT) cytotoxicity assay in Spodoptera frugiperda (Sf9) cells revealed that PcE1-, PcE7- or PcE9-expressing cells showed significantly higher cytoprotective capability than parental Sf9 cells against fenpropathrin, demonstrating that PcEs probably detoxify fenpropathrin. Moreover, gene silencing through the method of leaf-mediated dsRNA feeding followed by insecticide bioassay increased the mortalities of fenpropathrin-treated mites by 31% (PcE1), 27% (PcE7) and 22% (PcE9), respectively, after individual PcE gene dsRNA treatment. In conclusion, this study provides evidence that PcE1, PcE7 and PcE9 are functional genes mediated in fenpropathrin resistance in P. citri and enrich molecular understanding of CarEs during the resistance development of the mite.

Functional analysis of a chitinase gene during the larval-nymph transition in Panonychus citri by RNA interference.

Chitinases are hydrolytic enzymes that are required for chitin degradation and reconstruction in arthropods. In this study, we report a cDNA sequence encoding a putative chitinase (PcCht1) from the citrus red mite, Panonychus citri. The PcCht1 (564 aa) possessed a signal peptide, a conserver domain, and a chitin-binding domain. Structural and phylogenetic analyses found that PcCht1 had high sequence similarity to chitinases in Tetranychus urticae. Real-time quantitative PCR analyses showed that the transcript levels of PcCht1 peaked periodically in larval and nymph stages. Moreover, significant increase of PcCht1 transcript level in the larvae was observed upon the exposure of diflubenzuron. In contrast, exposures of the larvae to diflubenzuron resulted in the decreased chitin content. Furthermore, through a feeding-based RNA interference approach, we were able to reduce the PcCht1 transcript level by 59.7 % in the larvae, and consequently the treated larvae showed a very low molting rate compared with the control. Our results expanded the understanding of the important role of PcCht1 in the growth and development of P. citri.

Involvement of acid-sensing ion channel 1α in hepatic carcinoma cell migration and invasion.

An acidic microenvironment promotes carcinoma cell proliferation and migration. Acid-sensing ion channels (ASICs) are H(+), Ca(2+), and Na(+)-gated cation channels that are activated by changes in the extracellular pH, and ASIC1α may be associated with tumor proliferation and migration. Here, we investigated the role of ASIC1α in hepatocellular carcinoma (HCC) migration and invasion. The expression of ASIC1α was examined in 15 paired HCC and adjacent non-tumor tissues by immunohistochemistry. Reverse transcription (RT)-PCR and Western blotting were used to assess ASIC1α messenger RNA (mRNA) and protein expression in the HCC cell line SMMC-7721 cultured in different pH media or transfected with short hairpin RNA (shRNA) against ASIC1α. Cell migration ability was detected by wound healing and Transwell assays. ASIC1α expression was significantly higher in tumor tissues than in non-tumor tissues, and it was higher in HCC with postoperative metastasis than in that without metastasis. ASIC1α mRNA and protein expression was significantly higher in SMMC-7721 cells cultured at pH 6.5 than in those cultured at pH 7.4 and 6.0. shRNA-mediated silencing of ASIC1α significantly downregulated ASIC1α mRNA and protein expression compared with negative control or untransfected cells and inhibited HCC cell migration and invasion. ASIC1α is overexpressed in HCC tissues and associated with advanced clinical stage. A moderately acidic extracellular environment promoted ASIC1α expression, and silencing of ASIC1α expression inhibited the migration and invasion of HCC cells. Suppression of ASIC1α expression by RNAi attenuated the malignant phenotype of HCC cells, suggesting a novel approach for anticancer gene therapy.

[Non-covalent albumin conjugates of silicon (IV) phthalocyanines axially substituted with nucleoside: preparation and in vitro photodynamic activities].

The interactions of bovine serum albumin (BSA) with five novel silicon (N) phthalocyanines(SiPcl-5) axially modified by nucleosides (cytidine, 5-N-cytidine, methyl cytidine, uridine and methyl uridine) derivatives were studied by fluorescence spectroscopy. The results show that there are strong interactions between these silicon phthalocyanines and BSA with a binding constant of (4.90-83.18) x 10(5) mol(-1) x L. Therefore, the non-covalent BSA conjugate of bis(2', 3'-O-isopropyl-cytidine-oxy) phthalocyaninatosilicon(IV) (SiPc1) was further been prepared. The molar ratio of phthalocyanine to albumin was found to be 1:1 for the obtained SiPcl-BSA conjugate. The absorption spectra of SiPc1 and SiPc1-BSA in the visible region have no significant difference, both showing an Q-band maximum at about 686 nm. It indicates that the spectroscopic characteristics of SiPc1 are not affected by binding to albumin. The SiPcl-BSA conjugate exhibits high photodynamic activity against human hepatoma cell line HepG2 with an IC50 value of 3.0 x 10(-7) mol x L(-1). By comparsion, SiPc1-BSA has a higher photodynamic activity than SiPc1 (in PBS formation, IC50 = 7.0 x 10(-7) mol x L(-1)), which can be attributed to its higher cellular uptake.

[Synthesis and photodynamic anticancer activity of silicon phthalocyanine axially modified by nucleoside derivatives].

A new axially modified silicon phthalocyanine, di [5'-(2', 3'-O-isopropyl)-5-methyl cytidineoxy] silicon phthalocyanine (SiPcG), was prepared and characterized by 1H NMR and HRMS. This compound is essentially nonaggregated in N,N-dimethyformamide and 1% cremophor EL aqueous solution. It shows a Qband at 676 nm and fluorescence emission at 685 nm in DMF, and exhibits a Q-band at 679 nm and fluorescence emission at 689 nm in 1% cremophor EL aqueous solution. The SiPcG shows a high photodynamic activity against human hepatoma cells HepG2 with an IC50 value down to 7.8 x 10(-8) mol x L(-1). Fluorescence confocal microscopy study indicated that the SiPcG locates preferentially in the mitochondria of cells. The research results show that the SiPcG is highly potential as a new anti-cancer photosensitizer.

[Spectral properties, photodynamic anticancer activity and the interaction with albumin of silicon phthalocyanine axially modified by pyrimidine derivatives].

Photophysical photochemical properties, in vitro photodynamic anticancer activity and interaction with albumin of a new axially modified silicon phthalocyanine, i. e. di (2-amino-6-trifluoromethyl-4-pyrimidinyloxy) silicon phthalocyanine (SiPcF), were studied in the present paper. The Q band maximum absorption of SiPcF located at 686 nm with the molar absorption coefficient of 2.3 x 10(5) mol(-1) * L * cm(-1). The fluorescence emission of SiPcF peaked at 694 nm with a fluorescence quantum yield of 0.46. Its singlet oxygen quantum yield produced by photosensitization is 0.38, suggesting that SiPcF is an effective 1O2 photosensitizer. There is strong interaction between SiPcF and bovine serum albumin (BSA). The binding constant is 4.33 x 10(5) mol X L(-1) and the number of binding sites is 1. In vitro experiments show that SiPcF had a high photodynamic anticancer activity against human hepatoma cells HepG2 with an IC50 value down to 5 x 10(-7) mol X L(-1).

Acute brucellosis with myelodysplastic syndrome presenting as pancytopenia and Fever of unknown origin.

OBJECTIVE: To report a rare case of brucellosis with myelo-dysplastic syndrome (MDS). CLINICAL PRESENTATION AND INTERVENTION: A 70-year-old woman presented with pancytopenia and fever of unknown origin (FUO). The initial diagnosis was brucellosis; the woman was treated with doxycycline and rifampin against Brucella melitensis but was later diagnosed as suffering from MDS. She was immediately transferred to the Department of Hematology for further evaluation. CONCLUSION: This study highlights the rarity of brucellosis with MDS, and we recommend that brucellosis with MDS be considered in patients presenting with pancytopenia and FUO.

[Effect of electroacupuncture on the multifidus muscle in patients with lumbar disc herniation].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on lumbar dysfunction and multifidus muscle characteristics in patients with lumbar disc herniation (LDH). METHODS: Sixty patients with LDH were randomly divided into an observation group and a control group, 30 cases in each group. The patients in the control group were treated with symptomatic treatment. On the basis of the treatment of the control group, the patients in the observation group were treated with acupuncture at L3-S1 Jiaji (Ex-B 2) and Dachangshu (BL 25), and the ipsilateral L3 and L5 Jiaji point were connected with EA (continuous wave, frequency of 20 Hz, and the intensity was appropriate to the patient's tolerance). The needles were retained for 20 min, once every other day, and 10 times were taken as a course of treatment. A total of 2 courses of treatment was given. The modified Oswestry disability index (ODI) as well as the physical component summary (PCS) and mental component summary (MCS) of the 36-item short form health survey (SF-36) were compared between the two groups before and after treatment. Lumbar MRI was performed before and after treatment to measure the cross-sectional area (CSA), fatty infiltration (FI) and T2 values of multifidus muscle at the lower edge of L4 and L5 vertebral bodies. RESULTS: After treatment, the ODI, PCS and MCS socres in the two groups were improved compared with those before treatment (P<0.05), and the ODI and PCS socres in the observation group were better than those in the control group (P<0.05). After treatment, the FI and value of T2 in the observation group were lower than those before treatment (P<0.05), and lower than those in the control group (P<0.05). CONCLUSION: EA could improve lumbar dysfunction, relieve edema and fatty infiltration of multifidus muscle in patients with LDH.

Antioxidant Role of PcGSTd1 in Fenpropathrin Resistant Population of the Citrus Red Mite, Panonychus citri (McGregor).

The citrus red mite, Panonychus citri, a major citrus pest distributed worldwide, has evolved severe resistance to various classes of chemical acaricides/insecticides including pyrethroids. It is well known that the resistance to pyrethroids is mainly caused by point mutations of voltage-gated sodium channel gene in a wide range of pests. However, increasing number of evidences support that pyrethroids resistance might also be resulted from the integrated mechanisms including metabolic mechanisms. In this study, firstly, comparative analysis of RNA-seq data showed that multiple detoxification genes, including a GSTs gene PcGSTd1, were up-regulated in a fenpropathrin-resistant population compared with the susceptible strain (SS). Quantitative real time-PCR results showed that the exposure of fenpropathrin had an induction effect on the transcription of PcGSTd1 in a time-dependent manner. In vitro inhibition and metabolic assay of recombinant PcGSTd1 found that fenpropathrin might not be metabolized directly by this protein. However, its antioxidant role in alleviating the oxidative stress caused by fenpropathrin was demonstrated via the reversely genetic experiment. Our results provide a list of candidate genes which may contribute to a multiple metabolic mechanisms implicated in the evolution of fenpropathrin resistance in the field population of P. citri. Furthermore, during the detoxification process, PcGSTd1 plays an antioxidant role by detoxifying lipid peroxidation products induced by fenpropathrin.

Prediction of Lymph Node Metastases in Gastric Cancer by Serum APE1 Expression.

Aims: To investigate the functional role of serum Human apurinic/apyrimidinic endonuclease 1 (APE1) in prediction of lymph node metastasis in gastric cancer patients. Materials and methods: Serum samples were pre-operational collected from 86 patients with gastric cancer from Tianjin Medical University Cancer Institute and Hospital from March 2016 to August 2016. The serum of APE1 was measured by ELISA development kit and other CA242, CA724, CA199 and CEA levels by electrochemiluminescence assay. Results: The total of 86 patients with gastric cancer was classified into two groups (lymph node positive and negative groups). Using ELISA assay, we found out that the concentration of serum APE1 was higher in lymph node positive group than that of lymph node negative group. The receiver operating characteristic (ROC) curve was performed to analyze, indicating that area under the ROC curve of serum APE1 were better than those of each regular markers (CEA+CA199+CA242+CA724) or combination of these markers. Additionally, the APE1 overexpression was uncovered in tissue of gastric cancer patients with lymph nodes metastases, which is correlation with results of serum APE1. Conclusion: Serum APE1 was identified as a valuable marker for prediction of lymph node metastases in patients with gastric cancer.

Over-expression of ASIC1a promotes proliferation via activation of the ß-catenin/LEF-TCF axis and is associated with disease outcome in liver cancer.

Acid-sensing ion channels 1a (ASIC1a) has been reported to promote migration and invasion in liver cancer. However, the clinical significance and molecular mechanism of ASIC1a in liver cancer remain unknown. In the study, we found that ASIC1a is frequently up-regulated in liver cancer tissues. The over-expression of ASIC1a is associated with advanced clinical stage and poor prognosis. The pro-proliferative of ASIC1a is pH dependent. Knockout of ASIC1a by CRISPR/CAS9 inhibited liver cancer cell proliferation and tumorigenicity in vitro and in vivo through ß-catenin degradation and LEF-TCF inactivation. Our results indicated a potential diagnostic marker and chemotherapeutic target for liver cancer.

Identification of responsive proteins in Panonychus citri exposed to abamectin by a proteomic approach.

Abamectin is a microbial-derived pesticide widely used for control of agricultural pests. However, sustained use of abamectin has led to the development of resistance in some target species. Previous studies on arthropod resistance to abamectin have mainly used traditional biochemical and molecular approaches. To understand the responses of citrus red mite, Panonychus citri, exposed to abamectin, comparative proteomic analysis was conducted using two-dimensional electrophoresis (2-DE). A total of 26 distinct protein spots were present in response to abamectin exposure. Tandem mass spectrometry (MS/MS) identified 16 proteins that were mainly involved in energy metabolism and detoxification. Some remaining proteins were not identifiable, suggesting that they may be novel. The expression levels of transcripts associated with proteins were analyzed by quantitative reverse transcription PCR (qRT-PCR). Furthermore, to validate the proteomic data obtained in the present study, Western-blot experiment was performed and the expression of sHsp and PcE1 proteins were confirmed, respectively. BIOLOGICAL SIGNIFICANCE: The citrus red mite has developed resistance to many acaricides, including abamectin. In the current study, we used the proteomic approaches involving 2-DE, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF), and MS/MS to document changes in adult P. citri during 24h of abamectin exposure. Abamectin stress induced a total of 16 differentially regulated proteins. The proteomic results were validated in mRNA expression patterns using qRT-PCR. This is the first analysis of differentially expressed proteins in P. citri exposed to abamectin. The results help clarify the physiological mechanisms of P. citri responses to abamectin exposure.

Silicon(IV) phthalocyanines substituted axially with different nucleoside moieties. Effects of nucleoside type on the photosensitizing efficiencies and in vitro photodynamic activities.

A series of new silicon(IV) phthalocyanines (SiPcs) di-substituted axially with different nucleoside moieties have been synthesized and evaluated for their singlet oxygen quantum yields (ΦΔ) and in vitro photodynamic activities. The adenosine-substituted SiPc shows a lower photosensitizing efficiency (ΦΔ=0.35) than the uridine- and cytidine-substituted analogs (ΦΔ=0.42-0.44), while the guanosine-substituted SiPc exhibits a weakest singlet oxygen generation efficiency with a ΦΔ value down to 0.03. On the other hand, replacing axial adenosines with chloro-modified adenosines and purines can result in the increase of photogenerating singlet oxygen efficiencies of SiPcs. The formed SiPcs 1 and 2, which contain monochloro-modified adenosines and dichloro-modified purines respectively, appear as efficient photosensitizers with ΦΔ of 0.42-0.44. Both compounds 1 and 2 present high photocytotoxicities against HepG2 and BGC823 cancer cells with IC50 values ranging from 9nM to 33nM. The photocytotoxicities of these two compounds are remarkably higher than the well-known anticancer photosensitizer, chlorin e6 (IC50=752nM against HepG2 cells) in the same condition. As revealed by confocal microscopy, for both cell lines, compound 1 can essentially bind to mitochondria, while compound 2 is just partially localized in mitochondria. In addition, the two compounds induce cell death of HepG2 cells likely through apoptosis.

The first silicon(IV) phthalocyanine-nucleoside conjugates with high photodynamic activity.

A series of novel silicon(IV) phthalocyanines conjugated axially with different nucleoside moieties (uridine, 5-methyluridine, cytidine, and 5-N-cytidine derivatives) have been synthesized and evaluated for their photodynamic activities. The uridine-containing compound 1 exhibits the highest photocytotoxicity against HepG2 human hepatocarcinoma cells with an IC50 value as low as 6 nM, which can be attributed to its high cellular uptake and non-aggregated nature in the biological media. This compound shows high affinity toward the mitochondria of HepG2 cells and causes cell death mainly through apoptosis upon illumination. The result indicates that 1 is a highly promising photosensitizer for photodynamic therapy.

Diagnostic value of fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography in patients with fever of unknown origin.

BACKGROUND: While fever of unknown origin (FUO) remains a challenging problem in clinical practice, fluorine-18 fluorodeoxyglucose ((18)F-FDG) positron emission tomography/computed tomography (PET/CT) has been considered helpful in diagnosing its cause. The present study is set to evaluate the diagnostic value of PET/CT for patients with FUO. METHODS: We analyzed the records of 48 patients with FUO (34 men and 14 women; mean age of 57-year-old with a range between 24- and 82-year-old). The patients were examined by (18)F-FDG PET/CT and the results were compared to a final diagnosis that was established by additional procedures. RESULTS: A final diagnosis was established for 36 patients (75%). Among them, 15 patients had infectious diseases, 12 patients had malignancies, and 9 patients had non-infectious inflammatory diseases. Thirty-two abnormal PET/CT results correctly revealed the source of fever (true-positives). Abnormal PET/CT results were considered false-positives for 8 patients without diagnoses. Normal PET/CT results in 4 patients with no diagnoses were classified as true-negatives. Four patients with normal PET/CT results with diagnosed cause for FUO were considered false-negatives. Therefore, PET/CT had a positive predictive value of 80%, a negative predictive value of 50%, a sensitivity of 89%, and a specificity of 33% in patients with FUO. CONCLUSIONS: Our study demonstrated that FDG-PET/CT is a valuable imaging tool for the identification of the etiology in patients with FUO. The results suggest that this procedure may be considered as a second-line test, especially when conventional structural imaging was normal or unable to distinguish lesions from benign and malignant.