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SignificanceThe adult Drosophila mushroom body (MB) is one of the most extensively studied neural circuits. However, how its circuit organization is established during development is unclear. In this study, we provide an initial characterization of the assembly process of the extrinsic neurons (dopaminergic neurons and MB output neurons) that target the vertical MB lobes. We probe the cellular mechanisms guiding the neurite targeting of these extrinsic neurons and demonstrate that Semaphorin 1a is required in several MB output neurons for their dendritic innervations to three specific MB lobe zones. Our study reveals several intriguing molecular and cellular principles governing assembly of the MB circuit.

Professor LU Fan adheres to the principle in clinical practice, "the needling principle concentrated on regulating qi ". She takes the advantages of shallow needling technique of acupuncture in treatment of various diseases, e.g. exogenous disease, initial onset of disorder, chronic bi disorder, intractable diseases, disorder of yang nature, disorder of heat nature, thin body, pediatric diseases, disorders on the unilateral side of the body and acute diseases. Besides in compliance with classics, she has broadened the application scope of shallow needling technique of acupuncture and improved the clinical therapeutic effect.

Oxidative stress is closely related to the occurrence of depression. Acupuncture has been proved to be an effective method for treating depression. In order to explore the mechanism of the antidepressant effect of acupuncture, this study performed acupuncture prevention on chronic unpredictable mild stress (CUMS) depression model rats, and observed the effect of acupuncture on hippocampal oxidative stress and Nrf2 signaling pathway. Male SD rats were randomly divided into control group, CUMS group, acupuncture group, and fluoxetine group ( n = 10/group). Fluoxetine, a common antidepressant, was used as a positive control drug in this study. In the fluoxetine group, rats were given fluoxetine (2.1 mg/kg) intragastrically once a day for 28 days. The acupoints of Shangxing (GV23) and Fengfu (GV16) were applied in acupuncture group, once every other day for 14 times in total. Behavioral tests and biological detections were used to evaluate the effects of the interventions and the changes of factors related to oxidative stress, Nrf2 pathway, and neuronal apoptosis. The results showed that both acupuncture and fluoxetine could increase sugar preference rate in SPT and decrease immobility time in FST in depression model rats. It also significantly decreased oxidative stress products such as ROS and H 2 O 2 , and elevated the protein and mRNA expressions of Nrf2 and HO-1. From Nissl's staining, there were more abundant nerve cells in two intervention groups compared with CUMS group. Plus, acupuncture down-regulated the expression levels of Bax and caspase-3 and up-regulated the expression of Bcl-2. Our findings indicate that acupuncture improved depression-like behaviors of CUMS rats. And CUMS-induced depression-like behaviors in rats were related to oxidative stress and neuronal apoptosis in hippocampus. Acupuncture showed antidepressant effects in reducing oxidative stress products via regulating the Nrf2/HO-1 signaling pathway so that prevented neuronal apoptosis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Cheng, Li, Huang, Huang, Chen, Chen, Shen, Chen, Long and Meng.)

Selective hydrogenation plays an important role in the chemical industry and has a wide range of applications, including the production of fine chemicals and petrochemicals, pharmaceutical synthesis, healthcare product development, and the synthesis of agrochemicals. Pd-based catalysts have been widely applied for selective hydrogenation due to their unique electronic structure and ability to adsorb and activate hydrogen and unsaturated substrates. However, the exclusive and comprehensive summarization of the size, composition, and surface and interface effect of metal Pd on the performance for selective hydrogenation is still lacking. In this perspective, the research progress on selective hydrogenation using Pd-based catalysts is summarized. The strategies for improving the catalytic hydrogenation performance over Pd-based catalysts are investigated. Specifically, the effects of the size, composition, and surface and interfacial structure of Pd-based catalysts, which could influence the dissociation mode of hydrogen, the adsorption, and the reaction mode of the catalytic substrate, on the performance have been systemically reviewed. Then, the progress on Pd-based catalysts for selective hydrogenation of unsaturated alkynes, aldehydes, ketones, and nitroaromatic hydrocarbons is revealed based on the fundamental principles of selective hydrogenation. Finally, perspectives on the further development of strategies for chemical selective hydrogenation are provided. It is hoped that this perspective would provide an instructive guideline for constructing efficient heterogeneous Pd-based catalysts for various selective hydrogenation reactions., Competing Interests: The authors declare no competing financial interest., (© 2021 The Authors. Published by American Chemical Society.)

Proper development of mammalian skeletal muscle relies on precise gene expression regulation. Our previous studies revealed that muscle development is regulated by both mRNA and long non-coding RNAs (lncRNAs). Accumulating evidence has demonstrated that N 6 -methyladenosine (m 6 A) plays important roles in various biological processes, making it essential to profile m 6 A modification on a transcriptome-wide scale in developing muscle. Patterns of m 6 A methylation in lncRNAs in developing muscle have not been uncovered. Here, we reveal differentially expressed lncRNAs and report temporal m 6 A methylation patterns in lncRNAs expressed in mouse myoblasts and myotubes by RNA-seq and methylated RNA immunoprecipitation (MeRIP) sequencing. Many lncRNAs exhibit temporal differential expression, and m 6 A-lncRNAs harbor the consensus m 6 A motif "DRACH" along lncRNA transcripts. Interestingly, we found that m 6 A methylation levels of lncRNAs are positively correlated with the transcript abundance of lncRNAs. Overexpression or knockdown of m 6 A methyltransferase METTL3 alters the expression levels of these lncRNAs. Furthermore, we highlight that the function of m 6 A genic lncRNAs might correlate to their nearby mRNAs. Our work reveals a fundamental expression reference of m 6 A-mediated epitranscriptomic modifications in lncRNAs that are temporally expressed in developing muscle., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Xie, Tao, Diao, Li, Chen, Zhou, Hu, Hou, Lei, Xu, Chen, Peng, Zhang and Xiao.)

To evaluate the application of outcome indicators in randomized controlled trials(RCTs) concerning the treatment of tension-type headache(TTH) with traditional Chinese medicine(TCM) in recent five years, so as to provide a basis for the study of core outcome set(COS) for TCM intervention in TTH. The RCTs on TCM treatment of TTH in recent five years were systematically retrieved from CNKI, Wanfang, VIP, CBM, EMbase, PubMed, Cochrane Library, Web of Science, ClinicalTrials.gov and China Clinical Trial Registry. After literature screening, data extraction and evaluation of the risk of bias, the outcome indicators in the included RCTs were subjected to qualitative analysis. The preliminary search yielded 19 042 articles, and 10 983 were left after the elimination of duplication. Finally, 52 RCTs(48 in Chinese and 4 in English) were included for qualitative analysis. The outcome indicators of RCTs included in this study were classified into seven domains: TCM syndrome, symptom and sign, physical and chemical detection, quality of life, long-term prognosis, economic evaluation, and safety event. The findings demonstrated that headache characteristic index in the symptom and sign domain was the index with the highest reporting frequency and reporting rate. Seventeen RCTs used TCM syndrome score as the outcome indicator. Further analysis revealed that there existed such problems in research design as non-distinction between primary and secondary outcome indicators, great difference in the adopted measurement tools for outcome indicators, and the neglect of measurement time of outcome indicators. Moreover, the syndrome indicators reflecting TCM advantages, objective evaluation indicators, safety and health-economic indicators were lacking. These limitations have affected the quality and reliability of RCTs on TTH treatment with TCM. It is suggested that the efficacy and characteristics of TCM should be combined into current clinical research, and the COS in RCTs regarding TCM treatment of TTH should be established according to internationally recognized standard procedures.

Purpose: Breast cancer is a common malignant tumor in women with a poor prognosis. This study aimed to investigate angiogenesis subtypes of breast cancer and unveil the etiology and molecular features of breast cancer., Methods: Based on the angiogenesis gene set derived from AmiGO2, and breast cancer data in the Cancer Genome Atlas (TCGA), we define a novel cluster of angiogenesis subtypes for patients by consensus clustering. The gene regulation, immune landscape, molecular characteristics, and clinical features as well as enrichment pathways were explored in the angiogenesis subtypes of breast cancer., Results: Two angiogenesis subtypes were established through consensus clustering, among which subtype1 included 275 patients and subtype2 included 813 patients. A total of 643 differential expressed genes and 109 miRNAs were found between the two subtypes. The gene set enrichment analysis showed that the enriched hallmark pathways in subtype2 were related to the cancer tumorigenesis and breast cancer progression, including estrogen response early estrogen response late, epithelial-mesenchymal transition (EMT), especially angiogenesis. The mutant-allele tumor heterogeneity and tumor mutation burden of non-angiogenesis subtype were significantly higher than that in the angiogenesis subtype. The stroma score, immune score and ESTIMATE score were significantly higher in angiogenesis subtype, while the tumor purity in angiogenesis subtype was considerably lower. Finally, most immune checkpoints were expressed higher in the angiogenesis subtype., Conclusions: The omics analysis has established a novel angiogenesis subtype of breast cancer and identified the characteristics of the immune microenvironment and genomic alteration of breast cancer. Thus, this angiogenesis subtype might provide new evidence for inhibiting the progression and immunotherapy response in breast cancer.

This study aims to systematically evaluate the clinical efficacy and safety of Toutongning Capsules in the treatment of tension-type headache(TTH), so as to provide a corresponding basis for clinical treatment. Eight commonly used medical research databases and two clinical trial registration systems were retrieved with the time interval from the establishment of the database or system to November 2020. The randomized controlled trials of Toutongning Capsules in the treatment of TTH were screened out according to the pre-set criteria. The quality of the included papers was evaluated by the bias risk assessment tool in Cochrane Reviewers Handbook 6.1 and the data were statistically analyzed by RevMan v5.4 provided by Cochrane collaboration. A total of 13 studies were included and the quality of methodology was generally low. Meta-analysis showed that Toutongning Capsules assisted with western medicine therapy can effectively reduce the pain intensity(MD_(VAS)=-1.94,95%CI[-2.50,-1.38],P<0.000 01;MD_(NRS)=-0.83,95%CI[-0.86,-0.80],P<0.000 01), headache duration(SMD=-0.98,95%CI[-1.17,-0.79],P<0.000 01), headache frequency(MD=-1.01,95%CI[-1.16,-0.85],P<0.000 01), headache index(MD=-11.13,95%CI[-12.10,-10.16],P<0.000 01), anxiety and depression scale score(MD_(HAMA)=-4.02,95%CI[-6.58,-1.46],P=0.002;MD_(HAMD)=-2.67,95%CI[-4.04,-1.29],P=0.000 1), while Toutongning Capsules as monotherapy only reduced the headache score(MD=-2.24,95%CI[-2.97,-1.51],P<0.000 01). The available clinical studies demonstrate that Toutongning Capsules combined with western medicine in the treatment of TTH can improve the related outcome indicators, but the clinical safety and efficacy of Toutongning Capsules alone remain unclear. Due to the small number and low quality of the included studies, large-sample, multi-center, high-quality and strictly designed randomized controlled trials are still needed to verify the clinical efficacy in the future.

The efficacy of gastrodin as a Chinese herbal medicine extract in the treatment of tension-type headache has been confirmed. This paper systematically reviewed the efficacy and safety of gastrodin in the treatment of tension-type headache, aiming to provide a new choice for the treatment of this disease. In this study, four Chinese databases, four English databases and two trial registries were searched from the date of establishment to September 2020. The related randomized controlled trials(RCTs) were screened out according to the predetermined criteria. The bias risk assessment tool developed by Cochrane collaboration was used to evaluate the quality of the reports. RevMan 5.4.1 was used for Meta-analysis, and GRADE system for the evidence-based evaluation on the quality of outcome indicators. A total of 177 articles were retrieved and 8 articles were finally included for analysis, with a total sample size of 1 091 cases, which included 565 cases in the treatment group and 526 cases in the control group. The overall quality of included stu-dies was not high. The results of Meta-analysis are as follows:(1)In terms of headache frequency, gastrodin group was better than wes-tern medicine group(MD=-2.90, 95%CI[-3.76,-2.03], P<0.000 01).(2)In terms of number of abnormal blood vessels in TCD, gastrodin group was better than western medicine group(MD=-88.96, 95%CI[-102.36,-75.55], P<0.000 01).(3)In terms of effective rate, gastrodin group was better than western medicine group(RR=1.47, 95%CI[1.29, 1.68], P<0.000 01). The results of subgroup analysis are as follows:(1)Effective rate based on age, for the patients upper age limit 40-46 years old, gastro-din group was better than western medicine group(RR=1.69, 95%CI[1.50, 1.90], P<0.000 01); for the patients upper age limit 55-60 years old, gastrodin group was better than western medicine group(RR=1.27, 95%CI[1.16, 1.38], P<0.000 01).(2)Effective rate based on dosage form, both the gastrodin capsules and injection groups were better than western medicine group(RR_(capsules)=1.42, 95%CI[1.08, 1.88], P=0.01; RR_(injection)=1.50, 95%CI[1.26, 1.77], P<0.000 01). GRADE evaluation showed that the above outcomes had low quality of evidence. Only one article detailed the occurrence of adverse reactions and thus the present study cannot make a positive conclusion on the safety of gastrodin in the treatment of tension-type headache. The small number and low quality of the included reports affected the reliability of the results. In the future, more high-quality randomized controlled trails are needed to improve the evaluation on the efficacy and safety of gastrodin in the treatment of tension-type headache.

Background: Liquid biopsy is gaining increasing popularity in cancer screening and diagnosis. However, there is no relatively mature DNA isolation method or commercial kit available that is compatible with different LB sample types. This study developed a PAN-sample DNA isolation method (PAN method) for liquid biopsy samples., Methods: The PAN method has two key steps, including biosample-specific pretreatments for various LB sample types and high concentration guanidine thiocyanate buffer for lysis and denaturation procedure. Subsequently, the performance of PAN method was validated by a series of molecular analyses., Results: The PAN method was used to isolate DNA from multiple sample types related to LB, including plasma, serum, saliva, nasopharyngeal swab, and stool. All purified DNA products showed good quality and high quantity. Comparison of KRAS mutation analysis using DNA purified using PAN method versus QIAamp methods showed similar efficiency. Epstein-Barr virus DNA was detected via Q-PCR using DNA purified from serum, plasma, nasopharyngeal swab, and saliva samples collected from nasopharyngeal carcinoma patients. Similarly, methylation sequencing of swab and saliva samples revealed good coverage of target region and high methylation of HLA-DPB1 gene. Finally, 16S rDNA gene sequencing of saliva, swab, and stool samples successfully defines the relative abundance of microbial communities., Conclusions: This study developed and validated a PAN-sample DNA isolation method that can be used for different LB samples, which can be applied to molecular epidemiological research and other areas., (© 2021 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC.)

Cost-efficient and environmentally friendly treatment of hydraulic fracturing effluents is of great significance for the sustainable development of shale gas exploration. We investigated the synergistic effects of plant-microbial treatment of shale gas fracturing waste fluid. The results showed that illumination wavelength and temperature are direct drivers for microbial treatment effects of COD Cr and BOD 5 , while exhibit little effects on nitrogen compounds, TDS, EC, and SS removals as well as microbial species and composition. Plant-microbial synergism could significantly enhance the removal of pollutants compared with removal efficiency without plant enhancement. Additionally, the relative abundance and structure of microorganisms in the hydraulic fracturing effluents greatly varied with the illumination wavelength and temperature under plant-microbial synergism. 201.24 g water dropwort and 435 mg/L activated sludge with illumination of 450-495 nm (blue) at 25 °C was proved as the best treatment condition for shale gas fracturing waste fluid samples, which showed the highest removal efficiency of pollutants and the lowest algal toxicity in treated hydraulic fracturing effluents. The microbial community composition (36.73% Flavobacteriia, 25.01% Gammaproteobacteria, 18.55% Bacteroidia, 9.3% Alphaproteobacteria, 4.1% Cytophagia, and 2.83% Clostridia) was also significantly different from other treatments. The results provide a potential technical solution for improved treatment of shale gas hydraulic fracturing effluents.

Objective: Plasma Epstein-Barr virus (EBV) DNA is considered a biomarker for nasopharyngeal carcinoma (NPC). However, its long-term role in NPC development is unclear., Materials and Methods: A total of 1363 participants seropositive for EBV VCA-IgA and EBNA1-IgA in a community-based NPC screening program in southern China were tested for plasma EBV DNA levels by real-time qPCR between 2008 and 2015. New NPC cases were confirmed by active follow-up approach and linkage to local cancer registry through the end of 2016. Cox proportional hazards regression analysis was performed to calculate the hazard ratios (HRs) for NPC risk with plasma EBV DNA., Results: Thirty patients were newly diagnosed during a median 7.5 years follow-up. NPC incidence increased with the plasma EBV DNA load ranging from 281.46 to 10,074.47 per 100,000 person-years in participants with undetectable and ≥ 1000 copies/ml levels; the corresponding cumulative incidence rates were 1.73 and 50%. Furthermore, plasma EBV DNA loads conferred an independent risk for NPC development after adjustment for other risk factors, with HRs of 7.63 for > 3-999 copies/ml and 39.79 for ≥1000 copies/ml. However, the HRs decreased gradually after excluding NPC cases detected in the first 2 to 3 years and became statistically nonsignificant by excluding cases detected during the first 4 years., Conclusion: Elevated plasma EBV DNA can predict NPC risk over 3 years. Monitoring plasma EBV DNA can be used as a complementary approach to EBV serological antibody-based screening for NPC.

Despite evidence suggesting the utility of Epstein-Barr virus (EBV) markers to stratify individuals with respect to nasopharyngeal carcinoma (NPC) risk in NPC high-risk regions, no validated NPC risk prediction model exists. We aimed to validate an EBV-based NPC risk score in an endemic population undergoing screening for NPC. This prospective study was embedded within an ongoing NPC screening trial in southern China initiated in 2008, with 51 235 adult participants. We assessed the score's discriminatory ability (area under the receiver-operator-characteristics curve, AUC). A new model incorporating the EBV score, sex and family history was developed using logistic regression and internally validated using cross-validation. AUCs were compared. We also calculated absolute NPC risk combining the risk score with population incidence and competing mortality data. A total of 151 NPC cases were detected in 2008 to 2016. The EBV-based score was highly discriminating, with AUC = 0.95 (95% CI = 0.93-0.97). For 90% specificity, the score had 87.4% sensitivity (95% CI = 81.0-92.3%). As specificity increased from 90% to 99%, the positive predictive value increased from 2.4% (95% CI = 1.9-3.0%) to 12.5% (9.9-15.5%). Correspondingly, the number of positive tests per detected NPC case decreased from 272 (95% CI = 255-290) to 50 (41-59). Combining the score with other risk factors (sex, first-degree family history of NPC) did not improve AUC. Men aged 55 to 59 years with the highest risk profile had the highest 5-year absolute NPC risk of 6.5%. We externally validated the discriminatory accuracy of a previously developed EBV score in a high-risk population. Adding nonviral risk factors did not improve NPC prediction., (© 2020 UICC.)

Background: Colorectal cancer (CRC), one of the most common malignancies worldwide, is associated with poor survival and has a high mortality rate. Taxol is a chemotherapeutic agent that has been clinically applied as a first-line drug against diverse cancers. Yet, development of drug resistance has become the major challenge for anti-cancer treatments. F-box and WD40 domain protein 7 ( Fbxw7 ) is a known tumor suppressor which is frequently downregulated in cancers. However, the biological roles and mechanisms of Fbxw7 in Taxol resistance are still under investigation., Methods: We report that Fbxw7 is significantly inactivated in CRC tumors and cell lines compared with normal tissues and colon cells. Expressions of Fbxw7 and Nox1 were detected from human colon tumors and cells by qRT-PCR and Western blot. Glycolysis rate was assessed by glucose uptake and lactate product assay. Interactions between Fbxw7 and Nox1 were determined by co-immunoprecipitation (Co-IP). Chemosensitivity and resistance of colon cancer cells were determined by MTT assay and Annexin V-FITC assay., Results: Overexpression of Fbxw7 sensitized colon cancer cells to Taxol. Moreover, we observed a negative correlation between Fbxw7 and glucose metabolism. From the established Taxol-resistant (TR) cell line from HCT-116, Fbxw7 was found to be markedly downregulated in HCT-116 TR cells. We detected that NADPH oxidase 1 ( Nox1 ), a superoxide-generating NADPH oxidase, is negatively regulated by Fbxw7 . The Co-IP assay showed that Fbxw7 interacted with Nox1 , which was observed to be significantly upregulated in CRC tissues. Nox1 therefore promotes the Taxol resistance and glucose metabolism of colon cancer cells. Finally, rescue experiments demonstrated that the Fbxw7 -promoted Taxol sensitivity was partially through the Nox1 -glycolysis axis. Restoration of Nox1 in Fbxw7 -overexpressed TR colon cancer cells significantly recovered the Taxol resistance, which could be further overridden by glycolysis inhibition., Conclusions: Collectively, this study uncovered that targeting the Fbxw7 - Nox1 -glucose metabolism axis could be an effective strategy against chemoresistant colon cancer., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at http://dx.doi.org/10.21037/atm-21-2076). The authors have no conflicts of interest to declare., (2021 Annals of Translational Medicine. All rights reserved.)

Nodal lines are degeneracies formed by crossing bands in three-dimensional momentum space. Interestingly, these degenerate lines can chain together via touching points and manifest as nodal chains. These nodal chains are usually embedded in two orthogonal planes and protected by the corresponding mirror symmetries. Here, we propose and demonstrate an in-plane nodal chain in photonics, where all chained nodal lines coexist in a single mirror plane instead of two orthogonal ones. The chain point is stabilized by the intrinsic symmetry that is specific to electromagnetic waves at the Г point of zero frequency. By adding another mirror plane, we find a nodal ring that is constructed by two higher bands and links with the in-plane nodal chain. The nodal link in momentum space exhibits non-Abelian characteristics on a C 2 T - invariant plane, where admissible transitions of the nodal link structure are determined by generalized quaternion charges. Through near-field scanning measurements of bi-anisotropic metamaterials, we experimentally mapped out the in-plane nodal chain and nodal link in such systems.

A defect pyrochlore-type Sn 1.06 Nb 2 O 5.59 F 0.97 (SnNbOF) nano-octahedron is used as a redox-active support for fabricating Au@SnO 2 core-shell and SnO 2 quantum dots at room temperature without the use of organic species or foreign reducing reagents. Gold (Au) and SnO 2 components were obtained through an in situ redox reaction between the HAuCl 4 and reductive Sn 2+ ions incorporated in SnNbOF. The composition and morphology of the resulting nanocomposites (denoted as Au-SnNbOF) could be controlled by adjusting the Au/SnNbOF ratio. The Au-SnNbOF nanocomposites exhibited efficient photoactivities for methyl orange (MO) degradation under the visible light irradiation ( λ > 420 nm), during which the MO was almost completely degraded within 8 min. Among all the samples, the 5wt% Au-SnNbOF nanocomposite had the highest rate constant (0.43 min -1 ), which was 40 times higher than that of the blank SnNbOF., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Background: Increased risk and cancer-related mortality is observed in pancreatic cancer (PC) patients with diabetes mellitus (DM). Whether using metformin as glucose-lowering therapy can result in survival benefit in this group of patients is still unclear., Methods: A meta-analysis of 21 studies that including 38,772 patients was performed to investigate the association between metformin and overall survival in patients with PC and concurrent DM., Results: A significant survival benefit was observed in metformin treatment group compared with non-metformin group (hazard ratio [HR] = 0.83, 95% confidence interval [CI]: 0.74-0.91). These associations were observed in both subgroups of Asian countries (HR = 0.69, 95% CI: 0.60-0.79) and Western countries (HR = 0.86, 95% CI: 0.76-0.95), the former was more obvious. Survival benefit was gained for patients at early stage (HR = 0.75, 95% CI: 0.64-0.85) and mixed stage (HR = 0.81, 95% CI: 0.70-0.91), but not for patients at advanced stage (HR = 0.99, 95% CI: 0.74-1.24). Similarly, survival benefit was also observed in patients receiving surgery (HR = 0.82, 95% CI: 0.69-0.94) and comprehensive treatment (HR = 0.85, 95% CI: 0.77-0.93), but not in chemotherapy group (HR = 0.99, 95% CI: 0.67-1.30). No obvious benefit was suggested when pooled by time-varying COX model (HR = 0.94, 95% CI: 0.86-1.03)., Conclusions: These results suggest that metformin is associated with survival benefit in patients with PC and concurrent DM. Further randomized controlled trials and prospective studies with larger sample sizes are required to confirm our findings.

H 2 S is abundantly available in nature, and it is a common byproduct in industries. Molybdenum sulfides have been proved to be active in the catalytic decomposition of hydrogen sulfide (H 2 S) to produce hydrogen. In this study, density functional theory (DFT) calculations are carried out to explore the reaction mechanisms of H 2 S with MS 3 (M = Mo, W) clusters. The reaction mechanism of H 2 S with MoS 3 is roughly the same as that of the reaction with WS 3 , and the free-energy profile of the reaction with MoS 3 is slightly higher than that of the reaction with WS 3 . The overall driving forces (-Δ G ) are positive, and the overall reaction barriers (Δ G b ) are rather small, indicating that such H 2 productions are product-favored. MS 3 (M = Mo, W) clusters have clawlike structures, which have electrophilic metal sites to receive the approaching H 2 S molecule. After several hydrogen-atom transfer (HAT) processes, the final MS 4 ·H 2 (IM-4) complexes are formed, which could desorb H 2 at a relatively low temperature. The singlet product MS 4 clusters contain the singlet S 2 moiety, similar to the adsorbed singlet S 2 on the surface of sulfide catalysts. The theoretical results are compared with the experiments of heterogeneous catalytic decomposition of H 2 S by MoS 2 catalysts. Our work may provide some insights into the optimal design of the relevant catalysts., Competing Interests: The authors declare no competing financial interest., (Copyright © 2020 American Chemical Society.)

Background: Pulmonary malignant neoplasms have a high worldwide morbidity and mortality, so the study of these malignancies using microRNAs (miRNAs) has attracted great interest and enthusiasm. The aim of this study was to determine the clinical effect of hsa-microRNA-204-5p (miR-204-5p) and its underlying molecular mechanisms in non-small cell lung cancer (NSCLC)., Methods: Expression of miR-204-5p was investigated by real-time quantitative PCR (RT-qPCR). After data mining from public online repositories, several integrative assessment methods, including receiver operating characteristic (ROC) curves, hazard ratios (HR) with 95% confidence intervals (95% CI), and comprehensive meta-analyses, were conducted to explore the expression and clinical utility of miR-204-5p. The potential objects regulated and controlled by miR-204-5p in the course of NSCLC were identified by estimated target prediction and analysis. The regulatory network of miR-204-5p, with its target genes and transcription factors (TFs), was structured from database evidence and literature references., Results: The expression of miR-204-5p was downregulated in NSCLC, and the downtrend was related to gender, histological type, vascular invasion, tumor size, clinicopathologic grade and lymph node metastasis (P<0.05). MiR-204-5p was useful in prognosis, but was deemed unsuitable at present as an auxiliary diagnostic or prognostic risk factor for NSCLC due to the lack of statistical significance in meta-analyses and absence of large-scale investigations. Gene enrichment and annotation analyses identified miR-204-5p candidate targets that took part in various genetic activities and biological functions. The predicted TFs, like MAX, MYC, and RUNX1, interfered in regulatory networks involving miR-204-5p and its predicted hub genes, though a modulatory loop or axis of the miRNA-TF-gene that was out of range with shortage in database prediction, experimental proof and literature confirmation., Conclusions: The frequently observed decrease in miR-204-5p was helpful for NSCLC diagnosis. The estimated target genes and TFs contributed to the anti-oncogene effects of miR-204-5p.

In attempts to delay tumor progression after surgery or minimally invasive local treatments, multidisciplinary strategies have been broadly studied in patients with hepatocellular carcinoma (HCC). The objective of this present study was to evaluate the efficacy of autologous transplantations of cytokine-induced killer (CIK) cells as an adjuvant therapy for patients with HCC. A total of 264 patients with HCC were enrolled in this retrospective study. Of these patients, 165 received either CIK cell therapy alone or as adjuvant therapy to surgery, transcatheter arterial chemoembolization (TACE), or TACE-based comprehensive treatments (CT). The remaining 99 patients received only surgery or TACE. Kaplan-Meier analysis and the Chi-squared test were used to analyze the overall survival (OS), progression-free survival (PFS), and clinical characteristics of the patients in the different treatment subgroups. Kaplan-Meier analysis suggested that patients in the Surgery+CIK group had a significantly improved OS compared with those in the other three groups (P < 0.001). Furthermore, patients who developed a fever after the CIK cell treatments manifested a likely better OS (P = 0.028). Subgroup analysis indicated that patients in the Surgery+CIK group likely had an improved PFS but a similar OS compared with the patients in the Surgery-alone group (P = 0.055 for PFS, and P = 0.746 for OS). Further subgroup analysis showed that the OS in both the TACE+CIK and CT+CIK groups was prolonged significantly compared with that in the TACE-alone group (P = 0.015 and P = 0.018, respectively). However, similar OS was observed between the TACE+CIK and CT+CIK groups (P = 0.686). Autologous transplantation of CIK cells as an adjuvant therapy was associated with better survival for patients with HCC, especially for those who had also undergone TACE. A fever reaction might be a potential event for assessing the curative effect of the CIK treatment., Competing Interests: None.

In the current study, we proposed to explore the potential role and related signaling pathways of Homobox A10 (HOXA10) in non-small cell lung cancer (NSCLC). HOXA10 levels in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) were detected by qRT-PCR and the expression of HOXA10 was significantly up-regulated in the NSCLC tissue of all 55 pairs (P = 0.037). Overexpression of HOXA10 was closely correlated with the clinical stage of LUSC (P = 0.011). HOXA10 expression in RNA sequencing data based on 1, 077 cases exhibited concordant significant up-regulation in NSCLC, LUAD and LUSC (P < 0.001). In NSCLC, HOXA10 expression was closely correlated to patient T stage (P = 0.006). In LUAD, HOXA10 expression was compactly correlated to patient N stage (P = 0.02). Some of the microarrays from Gene Expression Omnibus (GEO) and ArrayExpress showed consistent over-expression of HOXA10 levels in NSCLCs. More importantly, the combined SMD value was 0.052 (95% CI: 0.29-0.75, P < 0.001) generated by meta-analysis from 47 datasets based on 4, 616 cases of NSCLC. The area under the curve (AUC) of SROC supported the over-expression of HOXA10 in NSCLC as being 0.88 (95% CI: 0.81-0.93), with sensitivity and specificity of 0.88 (95% CI: 0.81-0.93) and 0.56 (95% CI: 0.44-0.66), respectively. In addition, 111 co-expressed genes were collected from cBioPortal and enriched in "cell cycle", "cell adhesion molecules", "p53 signaling", and "adherens junction". Interestingly, an up-regulation trend of HOXA10 protein expression was also observed in NSCLC through tissue chips and immunohistochemistry. In conclusion, the overexpression of HOXA10 may play a pivotal role in the tumorigenesis of NSCLC, and this effect is observed more obviously in LUSC than in LUAD., Competing Interests: None.

Density functional theory (DFT) calculations have been performed to explore the gas-phase hydrolysis reaction of mononuclear thorium halide clusters ThX4 (X = F, Cl). We have found that the hydrolysis of ThCl4 is easier than that of ThF4. Furthermore, their hydrolysis reactions favor pathways of direct dehydration of Th(OH)4 instead of further hydrolysis of ThOX2. There are some differences between the hydrolysis of ThCl4 and that of MCl4 (M = Ti, Zr and Hf). The X-HY (X = F, Cl; Y = F, Cl and OH) hydrogen bonds play an important role in the hydrogen transfer process of the hydrolysis reaction. The differences in the steric effects and bonding may be important factors that are related to the disparities in the hydrolysis of the above-mentioned metal halides.

In this article, we have presented a computational analysis on the structure and bonding of [Pd 2 Sn 18 ] 4- and illustrated that it serves as an interesting example of OMO-UMO mutual delocalisation with two identical [PdE 9 ] 2- fragments. We have also illustrated the alternative roles that could be played by an [ME 9 ] 2- /[E 9 ] 2- fragment, a simple L-type donor and a lone-pair acceptor.

Aim: To evaluate the impact of recombinant Bacteroides fragilis enterotoxin-2 (BFT-2, or Fragilysin) on colorectal tumorigenesis in mice induced by azoxymethane/dextran sulfate sodium (AOM/DSS)., Methods: Recombinant proBFT-2 was expressed in Escherichia coli strain Rosetta (DE3) and BFT-2 was obtained and tested for its biological activity via colorectal adenocarcinoma cell strains SW-480. Seventy C57BL/6J mice were randomly divided into a blank (BC; n = 10), model (AD; n = 20), model + low-dose toxin (ADLT; n = 20, 10 μg), and a model + high-dose toxin (ADHT; n = 20, 20 μg) group. Mice weight, tumor formation and pathology were analyzed. Immunohistochemistry determined Ki-67 and Caspase-3 expression in normal and tumor tissues of colorectal mucosa., Results: Recombinant BFT-2 was successfully obtained, along with its biological activity. The most obvious weight loss occurred in the AD group compared with the ADLT group (21.82 ± 0.68 vs 23.23 ± 0.91, P < 0.05) and the ADHT group (21.82 ± 0.68 vs 23.57 ± 1.06, P < 0.05). More tumors were found in the AD group than in the ADLT and ADHT groups (19.75 ± 3.30 vs 6.50 ± 1.73, P < 0.05; 19.75 ± 3.30 vs 6.00 ± 2.16, P < 0.05). Pathology showed that 12 mice had adenocarcinoma and 6 cases had adenoma in the AD group. Five mice had adenocarcinoma and 15 had adenoma in the ADLT group. Four mice had adenocarcinoma and 16 had adenoma in the ADHT group. The incidence of colorectal adenocarcinoma in both the ADHT group and the ADHT group was reduced compared to that in the AD group ( P < 0.05, P < 0.05). The positive rate of Ki-67 in the ADLT group and the ADHT group was 50% and 40%, respectively, both of which were lower than that found in the AD group (94.44%, P < 0.05, P < 0.05). Caspase-3 expression in the ADLT group and the ADHT group was 45% and 55%, both of which were higher than that found in the BC group (16.67%, P < 0.05, P < 0.05)., Conclusion: Oral administration with lower-dose biologically active recombinant BFT-2 inhibited colorectal tumorigenesis in mice., Competing Interests: Conflict-of-interest statement: To the best of our knowledge, no conflict of interest exists.

Taking four wheat varieties developed by Northwest Institute of Plateau Biology, Chinese Academy of Sciences, as test materials, with the measurement of content of photosynthetic pigments, leaf area, fresh and dry mass of flag leaf, the PS II photochemistry efficiency of abaxial and adaxial surface of flag leaf and its adaptation to strong solar radiation during the period of heading stage in Xiangride region were investigated with the pulse-modulated in-vivo chlorophyll fluorescence technique. The results indicated that flag leaf angle mainly grew in horizontal state in Gaoyuan 314, Gaoyuan 363 and Gaoyuan 584, and mainly in vertical state in Gaoyuan 913 because of its smaller leaf area and larger width. Photosynthetic pigments were different among the 4 varieties, and positively correlated with intrinsic PS II photochemistry efficiencies (Fv/Fm). In clear days, especially at noon, the photosynthetic photoinhibition was more serious in abaxial surface of flag leaf due to directly facing the solar radiation, but it could recover after reduction of sunlight intensity in the afternoon, which meant that no inactive damage happened in PS II reaction centers. There were significant differences of PS II actual and maximum photochemical efficiencies at the actinic light intensity (ΦPS II and Fv'/Fm') between abaxial and adaxial surface, and their relative variation trends were on the contrary. The photochemical and non-photochemical quenching coefficients (qP and NPQ) had a similar tendency in both abaxial and adaxial surfaces. Although ΦPS II and qP were lower in adaxial surface of flag leaf, the Fv'/Fm' was significantly higher, which indicated that the potential PS II capture efficiency of excited energy was higher. The results demonstrated that process of photochemical and non-photochemical quenching could effectively dissipate excited energy caused by strong solar radiation, and there were higher adaptation capacities in wheat varieties natively cultivated in Qinghai-Tibetan Plateau area.

Density functional theory (DFT) and coupled cluster theory (CCSD(T)) calculations are carried out to investigate the electronic and structural properties of a series of bimetallic oxide clusters MW2O9(-/0) (M=V, Nb, Ta). Generalized Koopmans' theorem is applied to predict the vertical detachment energies (VDEs) and simulate the photoelectron spectra (PES). Theoretical calculations at the B3LYP level yield singlet and doublet ground states for the bimetallic anionic and neutral clusters, respectively. All the clusters present the six-membered ring structures with different symmetries, except that the TaW2O9(-) cluster shows a chained style with a penta-coordinated tantalum atom. Spin density analyses reveal oxygen radical species in all neutral clusters, consistent with their structural characteristics. Moreover, additional calculations are performed to study the oxidation reaction of CO molecule with the W3O9(+) cation and the isoelectronic VW2O9 cluster, and results indicate that the introduction of vanadium at tungsten site can efficiently improve the oxidation reactivity., (Copyright © 2013 Elsevier B.V. All rights reserved.)

We investigated the structures and bonding of two series of early transition-metal oxide clusters, M(2)O(n)(-) and M(2)O(n) (M = Nb, Ta; n = 5-7) using photoelectron spectroscopy (PES) and density-functional theory (DFT). The stoichiometric M(2)O(5) clusters are found to be closed shell with large HOMO-LUMO gaps, and their electron affinities (EAs) are measured to be 3.33 and 3.71 eV for M = Nb and Ta, respectively; whereas EAs for the oxygen-rich clusters are found to be much higher: 5.35, 5.25, 5.28, and 5.15 eV for Nb(2)O(6), Nb(2)O(7), Ta(2)O(6), and Ta(2)O(7), respectively. Structural searches at the B3LYP level yield triplet and doublet ground states for the oxygen-rich neutral and anionic clusters, respectively. Spin density analyses reveal oxygen radical, diradical, and superoxide characters in the oxygen-rich clusters. The M(2)O(7)(-) and M(2)O(7) clusters, which can be viewed to be formed by M(2)O(5)(-/0) + O(2), are utilized as molecular models to understand dioxygen activation on M(2)O(5)(-) and M(2)O(5) clusters. The O(2) adsorption energies on the stoichiometric M(2)O(5) neutrals are shown to be surprisingly high (1.3-1.9 eV), suggesting strong capabilities to activate O(2) by structural defects in Nb and Ta oxides. The PES data also provides valuable benchmarks for various density functionals (B3LYP, BP86, and PW91) for the Nb and Ta oxides.

Over-activation of the local chymase-angiotensin II (Ang II) system has a dominant role in diabetic cardiomyopathy. Astragalus polysaccharides (APS) are used in traditional Chinese medicine to boost immunity. In this study, we investigated the effects of APS treatment on cardiac function, myocardial collagen expression, cardiac ultrastructure, cardiac matrix metalloproteinase (MMP) activity, levels of plasma glycosylated serum protein (GSP), and myocardial enzymes, and the expression of Ang II, chymase, and angiotensin-converting enzyme (ACE) in the diabetic hamster myocardium. Diabetes was induced by a single injection of streptozotocin (60 mg/kg ip). The experimental groups consisted of normal control (n = 15), diabetic (n = 15), insulin-treated diabetic (n = 15, NPH 1-2 U/day ip), and APS-treated diabetic (n = 30, APS 1-2 g/kg/day orally for 10 weeks) hamsters. Diabetic hamsters treated with insulin or APS exhibited significantly decreased blood glucose, plasma GSP, and myocardial enzymes, as well as improvements in cardiac function and cardiac ultrastructure. Compared with insulin treatment, APS treatment significantly reduced myocardial collagen (type I and III) expression and lowered cardiac MMP-2 activity, myocardial Ang II levels, myocardial chymase expression, and p-ERK1/2 kinase expression. In diabetic hamsters, myocardial ACE expression and plasma Ang II levels was not altered by insulin or APS treatment. These results indicate that treatment of diabetic hamsters with APS inhibited the local chymase-Ang II system and improved markers of diabetic cardiomyopathy.

The electronic and structural properties of a series of triniobium oxide clusters, Nb(3)O(n)(-) and Nb(3)O(n) (n = 3-8), are investigated using photoelectron spectroscopy (PES) and density functional theory (DFT) calculations. PES spectra are obtained for Nb(3)O(n)(-) (n = 3-8) at various photon energies and are used to compare with the DFT calculations. A trend of sequential oxidation is observed as a function of O content until Nb(3)O(8)(-), reaching the highest oxidation state of Nb. DFT calculations are performed to search for the lowest energy structures for both the anionic and neutral clusters. For Nb(3)O(3)(-), the three O atoms are shown to prefer the bridging sites of a triangular Nb(3), leading to two nearly degenerate cyclic structures of C(s) symmetry. The next three O atoms from n = 4-6 each occupy a terminal site directly bonded to Nb, resulting in a symmetric Nb(3)O(6)(-) with C(3v) symmetry and a low-lying isomer of C(s) symmetry. The seventh O atom is bonded to two Nb atoms forming a double bridge, whereas the eighth O atom is bonded to a terminal site so that in Nb(3)O(8)(-) each Nb atom reaches its maximum oxidation state of +5. The structures and electronic states for the triniobium oxide clusters are significantly different from the corresponding tritantalum oxide clusters, in particular, for Nb(3)O(3)(-), Nb(3)O(5)(-), and Nb(3)O(7)(-). Molecular orbital analyses are performed to elucidate the chemical bonding and the electronic and structural evolution in these triniobium oxide clusters.

Density functional theory (DFT) calculations are carried out to investigate the electronic and structural properties of a series of tetratungsten oxide clusters, W(4)O(n)(-/0) (n = 10-13). Generalized Koopmans' theorem is applied to predict the vertical detachment energies and simulate the photoelectron spectra (PES). A large energy gap (approximately 2.9 eV) is observed for the stoichiometric W(4)O(12) cluster, which reaches the bulk value. The calculations suggest that W(4)O(12)(-/0) have the planar eight-membered ring structures, in which each tungsten atom is tetrahedrally coordinated with two bridging O atoms and two terminal O atoms. W(4)O(10)(-/0) and W(4)O(11)(-) can be viewed as removing two and one terminal O atoms from W(4)O(12)(-/0), respectively. The W(4)O(11) neutral is an interesting species, which possesses the pentabridged structure. We show that W(4)O(11)(-) contains a localized W(3+) site, which can readily react with O(2) to form the W(4)O(13)(-) cluster, whereas the corresponding neutral W(4)O(13) can be viewed as replacing a terminal oxygen in W(4)O(12) by a peroxo O(2) unit. Molecular orbital analyses are performed to analyze the chemical bonding in the tetratungsten oxide clusters and to elucidate their electronic and structural evolution.

Objective: To evaluate the therapeutic effect of a new recombinant immunotoxin mMIP-1alpha-DT390 on experimental autoimmune encephalomyelitis (EAE)., Methods: EAE was induced in the low-sensitive strain C57BL/6 mice with intraperitoneal injection of myelin basic protein (MBP) to simulate the human disease multiple sclerosis, followed by intramuscular injection of cationic liposome carrying the plasmid DNA SRalpha-mMIP-1alpha-DT390 in the leg muscle to elicit resistance to EAE development. The mice were then examined daily for clinical signs of EAE by an observer blind to the treatment protocol. For immunohistochemistry the mice were anesthetized and perfused with sterile PBS and paraformaldehyde, and the cerebrum, cerebellum, medulla and spinal cord were removed for preparation of serial sections. The mononuclear cells (MNCs) from the EAE mouse spleens were prepared for three-color flow cytometry analysis of the surface markers with appropriate antibodies following the BD Pharmingen cytokine staining protocol., Results: EAE model was successfully established by active MBP immunization in C57BL/6 mice. Administration of the immunotoxin mMIP-1alpha-DT390 significantly delayed the disease onset and lowered the mean clinical score for EAE as compared with the control mice. Immunohistochemistry demonstrated much less CCR5(+) infiltrating cells in the central nervous system in mMIP-1alpha-DT390-treated mice than in the control. The treatment also eliminated reactive T cells in the periphery blood without affecting the number of B cells., Conclusion: The immunotoxin mMIP-1alpha-DT390 can attenuate the disease activity of EAE in mice, suggesting its potential use in the treatment of other autoimmune disorders.

Aim: To construct a novel eukaryotic expression plasmid including the recombinant immunotoxin DT390-mRantes and treat experimental autoimmune encephalomyelitis (EAE) in mice., Methods: EAE in C57BL/6 mice were induced by the extracted MBP. The mRantes fragment was inserted into the eukaryotic expression plasmid SRalpha containing DT390. Then cationic liposome-embedded plasmid DNA was injected into the muscles of the hind-limbs in mice. The effect of DT390-mRantes was evaluated by observing clinical symptoms, pathological changes of brain, relative cytokine of peripheral blood, and the proportion of T cells and B cells., Results: The recombinant immunotoxin DT390-mRantes was successfully constructed. Compared the mice in treated group with those in untreated group the clinical symptoms of EAE were alleviated, the infiltration of inflammatory cells were decreased, the IFN-gamma level was fallen, and the ratio of T/B cells was decreased., Conclusion: The recombinant immunotoxin DT390-mRantes has distinct effects on EAE in mice, which may be used for beneficial reference to the therapy of MS.

Objective: To construct a novel recombinant immunotoxin (IT) expression vector by fusing mouse macrophage inflammatory protein-1alpha gene (mMIP-1alpha) into a truncated diphtheria toxin gene (DT390), and examine the expression of mMIP-1alpha-DT390 fusion protein in NIH3T3 cells., Methods: mMIP-1alpha cDNA was cloned from mouse liver tissue through reverse transcription-polymerase chain reaction (RT-PCR), and inserted in the expression plasmid SRalpha, that includes the DT390 gene, to form the recombinant vector SRa-mMIP-1alpha-DT390. The positive recombinant plasmid was identified by PCR, the restriction endonucleases digestion and DNA sequencing, and then by liposome protocol, the identified positive plasmid was transferred into NIH3T3 cells for observing the fusion protein expression by immunofluorescence, with detecting the activities of the immunotoxin in vitro through MTT., Results: We have successfully constructed a recombinant immunotoxin expression vector named as SRalpha-mMIP-1alpha-DT390. The immunofluorescence photograph sourced from fluorescence immunocytochemical method showed that the fusion gene could express in the cytomembrane and cytoplasm of NIH3T3 cells. In the bioactivity detection assay, the supernatant of the transfected cell culture was observed to have the obvious cytotoxic activity to activated T-lymphocyte., Conclusion: The SRalpha-mMIP-1alpha-DT390 recombinant immunotoxin expression vector will provide the basis of studying the targeted cytotoxic activity to inflammatory cells, and may have some potential value for clinical application.

Objective: To investigate the expression of galectin-3 in benign and malignant thyroid nodules., Methods: Galectin-3 expression was assayed with immunohistochemical and RT-PCR methods in thyroid specimens., Results: The majority of the specimens of thyroid carcinomas expressed galectin-3 with immunohistochemical and RT-PCR methods (89.7% and 91.7%) and the expression was significantly in different histological types. Papillary and follicular thyroid malignancies showed high expression of galectin-3 (100% positive). Comparing the specimens of thyroid carcinomas with metastases and those without metastases, the expression of galectin-3 was significantly higher in the former than in the latter. Among 25 specimens of benign adenomas and 13 specimens of nodular hyperplasia, only 3 were weakly positive and other were negative. Neither 10 specimens of normal thyroid tissue nor 25 specimens of paracarcinomatous thyroid tissue expressed galectin-3., Conclusion: Galectin-3 may be used as a marker for differentiating benign and malignant thyroid nodules.

Aim: To construct a new recombinant eukaryotic expression plasmid of murine IFN-gamma-inducible protein 10 (mIP10) gene and a truncated diphtheria toxin (DT390) gene and investigate its biological function., Methods: mIP10 cDNA was cloned from murine liver tissue by RT-PCR and was inserted into eukaryotic expression plasmid SRalpha containing DT390 gene to construct plasmid SRalpha-DT390-mIP10. After being identified with restriction endonucleases digestion analysis and DNA sequencing, the recombinant plasmid was transfected into NIH3T3 cells through PolyFect. The expression of recombinant plasmid was detected by immunofluorescence assay and the bioactivity of mIP10-DT390 in cultured supernatant of the transfected NIH3T3 cells was detected by MTT colorimetry., Results: A new recombinant immunotoxin expression plasmid SRalpha-DT390-mIP10 was constructed successfully and was expressed in NIH3T3 cells. The expressed DT390-mIP10 killed the activated T cells effectively., Conclusion: A new recombinant eukaryotic expression plasmid SRalpha-DT390-mIP10 has been constructed and expressed successfully, which is useful in further studying the treatment of cancer and autoimmune diseases.

Objective: To obtain the recombinant fusion proteins GST-Rta185 and GST-Rta150 from EB virus and prepare two Rta protein specific polyclonal antibodies, respectively., Methods: Plasmids pGEX-R1501, pGEX-R1851 and pGEX-5X-3 were separately transformed into Escherichia coli BL21 (DE3). Expressions of the recombinant proteins R150-GST, R185-GST and free GST were induced by 0.1 mmol/L IPTG in LB medium. The expressed proteins were purified from lysates with Glutathione Sepharose 4B. Purified proteins were mixed with Freund's adjuvant and then were used to immunize rabbits., Results: High levels of expression of target proteins were detected in the lysates and the purified proteins were obtained by affinity chromatography with Glutathione Sepharose 4B. Western blot and ELISA analysis suggested that the polyclonal antibodies against GST-R185 and GST-R150 were specific., Conclusion: The antiserums have good specificity. They are important for the research on Rta fusion proteins from EB virus and for the diagnosis or treatment of EB virus associated diseases.

Unlabelled: This prospective study investigates the relationship between glucose transporter-1 (Glut-1) expression and PET images using (18)F-FDG and its uptake and compares them with the tumor status (primary vs. recurrent or persistent), initial grade of histologic differentiation, and International Federation of Gynecologic Obstetrics (FIGO) staging for cervical cancer patients., Methods: A dual-phase (18)F-FDG PET scan was performed on 51 participants within the 2 wk before surgery or biopsy. (18)F-FDG uptake was quantified by calculating standardized uptake values (SUVs). After (18)F-FDG PET scanning, 51 histologically proven squamous cell carcinoma specimens were examined to determine their degree of differentiation, using hematoxylin and eosin staining, and the expression of Glut-1 by an immunohistochemical stain. Twenty normal cervical and 20 cervical intraepithelial neoplasia (CIN) sets of tissue were also used to compare the results of Glut-1 expression in these tissues. The expression of Glut-1 was the product of (the intensity [with grades 0-3, defined qualitatively]) with (percentages of the lesion area that were positive). The results of Glut-1 expression were analyzed in combination with the SUVs (SUV1 was that at 40 min and SUV2 was that at 3 h), tumor status, initial cell differentiation, and FIGO staging., Results: Significant overexpression of Glut-1 was noted in 48 of the 51 (94.1%) cancer specimens. None or only minimal expression of Glut-1 was observed in basal layers of normal and CIN tissues. Significant positive correlation was observed between Glut-1 expression and the SUVs in cervical cancer specimens (r = 0.74, P < 0.000 for SUV1 and r = 0.65, P < 0.000 for SUV2). In recurrent or persistent tumor, tumor size was significantly associated with both Glut-1 expression (r = 0.508, P = 0.011) and SUV1 (r = 0. 456, P = 0.025). For recurrent or persistent tumor, only SUV1 reached statistical significance when compared with lymph node metastasis (P = 0.0226)., Conclusion: Glut-1 expression was related to (18)F-FDG uptake in cervical cancer patients. Recurrent or persistent cervical cancer tumor had significantly higher Glut-1 expression than metastatic lymph nodes. The values of SUV and the expression of Glut-1 did not correlate with the initial grade of histologic differentiation and FIGO staging.