Involvement of circRNA Regulators MBNL1 and QKI in the Progression of Esophageal Squamous Cell Carcinoma.

OBJECTIVES: To investigate the role of circRNA regulators MBNL1 and QKI in the progression of esophageal squamous cell carcinoma. BACKGROUND: MBNL1 and QKI are pivotal regulators of pre-mRNA alternative splicing, crucial for controlling circRNA production - an emerging biomarker and functional regulator of tumor progression. Despite their recognized roles, their involvement in ESCC progression remains unexplored. METHODS: The expression levels of MBNL1 and QKI were examined in 28 tissue pairs from ESCC and adjacent normal tissues using data from the GEO database. Additionally, a total of 151 ESCC tissue samples, from stage T1 to T4, consisting of 13, 43, 87, and 8 cases per stage, respectively, were utilized for immunohistochemical (IHC) analysis. RNA sequencing was utilized to examine the expression profiles of circRNAs, lncRNAs, and mRNAs across 3 normal tissues, 3 ESCC tissues, and 3 pairs of KYSE150 cells in both wildtype (WT) and those with MBNL1 or QKI knockouts. Transwell, colony formation, and subcutaneous tumorigenesis assays assessed the impact of MBNL1 or QKI knockout on ESCC cell migration, invasion, and proliferation. RESULTS: ESCC onset significantly altered MBNL1 and QKI expression levels, influencing diverse RNA species. Elevated MBNL1 or QKI expression correlated with patient age or tumor invasion depth, respectively. MBNL1 or QKI knockout markedly enhanced cancer cell migration, invasion, proliferation, and tumor growth. Moreover, the absence of either MBNL1 or QKI modulated the expression profiles of multiple circRNAs, causing extensive downstream alterations in the expression of numerous lncRNAs and mRNAs. While the functions of circRNA and lncRNA among the top 20 differentially expressed genes remain unclear, mRNAs like SLCO4C1, TMPRSS15, and MAGEB2 have reported associations with tumor progression. CONCLUSIONS: This study underscores the tumor-suppressive roles of MBNL1 and QKI in ESCC, proposing them as potential biomarkers and therapeutic targets for ESCC diagnosis and treatment.

Baicalein triggers ferroptosis in colorectal cancer cells via blocking the JAK2/STAT3/GPX4 axis.

Colorectal cancer (CRC) is a prevalent form of gastrointestinal malignancy with challenges in chemotherapy resistance and side effects. Effective and low toxic drugs for CRC treatment are urgently needed. Ferroptosis is a novel mode of cell death, which has garnered attention for its therapeutic potential against cancer. Baicalein (5, 6, 7-trihydroxyflavone) is the primary flavone extracted from the dried roots of Scutellaria baicalensis that exhibits anticancer effects against several malignancies including CRC. In this study, we investigated whether baicalein induced ferroptosis in CRC cells. We showed that baicalein (1-64 µM) dose-dependently inhibited the viability of human CRC lines HCT116 and DLD1. Co-treatment with the ferroptosis inhibitor liproxstatin-1 (1 µM) significantly mitigated baicalein-induced CRC cell death, whereas autophagy inhibitor chloroquine (25 µM), necroptosis inhibitor necrostatin-1 (10 µM), or pan-caspase inhibitor Z-VAD-FMK (10 µM) did not rescue baicalein-induced CRC cell death. RNA-seq analysis confirmed that the inhibitory effect of baicalein on CRC cells is associated with ferroptosis induction. We revealed that baicalein (7.5-30 µM) dose-dependently decreased the expression levels of GPX4, key regulator of ferroptosis, in HCT116 and DLD1 cells by blocking janus kinase 2 (JAK2)/STAT3 signaling pathway via direct interaction with JAK2, ultimately leading to ferroptosis in CRC cells. In a CRC xenograft mouse model, administration of baicalein (10, 20 mg/kg, i.g., every two days for two weeks) dose-dependently inhibited the tumor growth with significant ferroptosis induced by inhibiting the JAK2/STAT3/GPX4 axis in tumor tissue. This study demonstrates that ferroptosis contributes to baicalein-induced anti-CRC activity through blockade of the JAK2/STAT3/GPX4 signaling pathway, which provides evidence for the therapeutic application of baicalein against CRC.

Observation of Robust and Long-Ranged Superperiodicity of Electronic Density Induced by Intervalley Scattering in Graphene/Transition Metal Dichalcogenide Heterostructures.

Two-dimensional (2D) h-BN and transition metal dichalcogenides (TMDs) are widely used as substrates of graphene because they are insulating, atomically flat, and without dangling bonds. Usually, it is believed that such insulating substrates will not affect the electronic properties of graphene, especially when the moiré pattern generated between them is quite small. Here, we present a systematic study of the electronic properties of graphene/TMD heterostructures with the period of the moiré pattern <1 nm, and our results reveal an unexpected sensitivity of electronic properties in graphene to the 2D insulating substrates. We demonstrate that there is a robust and long-ranged superperiodicity of electronic density in graphene, which arises from the scattering of electrons between the two valleys of graphene in the graphene/TMD heterostructures. By using scanning tunneling microscope and spectroscopy, three distinct atomic-scale patterns of the electronic density are directly imaged in every graphene/TMD heterostructure.

Chemotherapy-induced metastasis: molecular mechanisms and clinical therapies.

Chemotherapy, the most widely accepted treatment for malignant tumors, is dependent on cell death induced by various drugs including antimetabolites, alkylating agents, mitotic spindle inhibitors, antitumor antibiotics, and hormonal anticancer drugs. In addition to causing side effects due to non-selective cytotoxicity, chemotherapeutic drugs can initiate and promote metastasis, which greatly reduces their clinical efficacy. The knowledge of how they induce metastasis is essential for developing strategies that improve the outcomes of chemotherapy. Herein, we summarize the recent findings on chemotherapy-induced metastasis and discuss the underlying mechanisms including tumor-initiating cell expansion, the epithelial-mesenchymal transition, extracellular vesicle involvement, and tumor microenvironment alterations. In addition, the use of combination treatments to overcome chemotherapy-induced metastasis is also elaborated.

The Histone H3K4 Demethylase JMJ16 Represses Leaf Senescence in Arabidopsis.

Leaf senescence is governed by a complex regulatory network involving the dynamic reprogramming of gene expression. Age-dependent induction of senescence-associated genes (SAGs) is associated with increased levels of trimethylation of histone H3 at Lys4 (H3K4me3), but the regulatory mechanism remains elusive. Here, we found that JMJ16, an Arabidopsis (Arabidopsis thaliana) JmjC-domain containing protein, is a specific H3K4 demethylase that negatively regulates leaf senescence through its enzymatic activity. Genome-wide analysis revealed a widespread coordinated upregulation of gene expression and hypermethylation of H3K4me3 at JMJ16 binding genes associated with leaf senescence in the loss-of-function jmj16 mutant as compared with the wild type. Genetic analysis indicated that JMJ16 negatively regulates leaf senescence, at least partly through repressing the expression of positive regulators of leaf senescence, WRKY53 and SAG201 JMJ16 associates with WRKY53 and SAG201 and represses their precocious expression in mature leaves by reducing H3K4me3 levels at these loci. The protein abundance of JMJ16 gradually decreases during aging, which is correlated with increased H3K4me3 levels at WRKY53 and SAG201, suggesting that the age-dependent downregulation of JMJ16 is required for the precise transcriptional activation of SAGs during leaf senescence. Thus, JMJ16 is an important regulator of leaf senescence that demethylates H3K4 at SAGs in an age-dependent manner.

Tunable Sample-Wide Electronic Kagome Lattice in Low-Angle Twisted Bilayer Graphene.

Overlaying two graphene layers with a small twist angle θ can create a moiré superlattice to realize exotic phenomena that are entirely absent in a graphene monolayer. A representative example is the predicted formation of localized pseudo-Landau levels (PLLs) with kagome lattice in tiny-angle twisted bilayer graphene (TBG) with θ<0.3° when the graphene layers are subjected to different electrostatic potentials. However, this was shown only for the model of rigidly rotated TBG, which is not realized in reality due to an interfacial structural reconstruction. It is believed that the interfacial structural reconstruction strongly inhibits the formation of the PLLs. Here, we systematically study electronic properties of the TBG with 0.075°≤θ<1.2° and demonstrate, unexpectedly, that the PLLs are quite robust for all the studied TBG. The structural reconstruction suppresses the formation of the emergent kagome lattice in the tiny-angle TBG. However, for the TBG around the magic angle, the sample-wide electronic kagome lattices with tunable lattice constants are directly imaged by using a scanning tunneling microscope. Our observations open a new direction to explore exotic correlated phases in moiré systems.

JMJ27-mediated histone H3K9 demethylation positively regulates drought-stress responses in Arabidopsis.

Dimethylation of histone H3 at lysine 9 (H3K9me2) is associated with heterochromatinization and transcriptional gene silencing in plants. The activation of sets of genes by drought stress is correlated with reduced H3K9me2 levels, but the role of H3K9 methylation in the regulation of drought-stress responses remains elusive. Here, we show that the Jumonji domain-containing H3K9 demethylase JMJ27 positively regulates drought-stress responses through its histone demethylase activity. RNA-seq analysis identified JMJ27-regulated genes, including positive regulators of drought stress GALACTINOL SYNTHASE 2 (GOLS2) and RESPONSE TO DESICCATION 20 (RD20). Genetic analysis demonstrated that JMJ27 positively regulates drought-stress responses at least partly through GOLS2 and RD20. JMJ27 directly associated with GOLS2 and RD20, and protected these loci from silencing by reducing H3K9me2 levels under normal conditions. REGULATORY PARTICLE NON-ATPASE 1a (RPN1a), a subunit of the 26S proteasome, interacted with JMJ27 and negatively regulated JMJ27 accumulation. Drought stress diminished RPN1a abundance, resulting in increased JMJ27 abundance. The drought stress-promoted occupancy of JMJ27 at GOLS2 and RD20 chromatin may reinforce their transcriptional induction by locally reducing the H3K9me2 levels. These results indicate that the RPN1a-JMJ27 module precisely regulates dynamic H3K9me2 deposition plasticity, ensuring proper adaptation to drought stress in Arabidopsis.

Oscillations of the Spacing between van Hove Singularities Induced by sub-Ångstrom Fluctuations of Interlayer Spacing in Graphene Superlattices.

Physical properties of two-dimensional van der Waals (vdWs) structures depend sensitively on both stacking orders and interlayer interactions. Yet, in most cases studied to date, the interlayer interaction is considered to be a "static" property of the vdWs structures. Here we demonstrate that applying a scanning tunneling microscopy (STM) tip pulse on twisted bilayer graphene (TBG) can induce sub-Ångstrom fluctuations of the interlayer separation in the TBG, which are equivalent to dynamic vertical external pressure of about 10 GPa on the TBG. The sub-Ångstrom fluctuations of the interlayer separation result in large oscillations of the energy separations between two van Hove singularities (VHSs) in the TBG. The period of the oscillations of the VHSs spacing is extremely long, about 500-1000 sec, attributing to tip-induced local stress in the atomic-thick TBG. Our result provides an efficient method to tune and measure the physical properties of the vdWs structures dynamically.

The development of a Chinese Healthy Eating Index for School-age Children and its Application in children from China Health and Nutrition Survey.

This study aimed to develop a Chinese Healthy Eating Index for School-age Children (CHEI-SC), apply it in the 2011 China Health and Nutrition Survey (CHNS) to assess dietary quality, and compared it with our former developed index named CHEI. Data of 3-day 24-hour diet recalls and household food inventory survey from 1600 school-age children in CHNS-2011 were used to develop the CHEI-SC, using the methods of standard portion size, energy-density-based approach, and least restrictive approach. The CHEI-SC included 19 components with a total score (T-score) ranging from 0 to 100. The investigated children had a median score of 49.6. Children with a higher T-score were more likely to have higher social economic status (SES), higher level of urbanisation, fewer family size, and regularly attending school. The CHEI-SC was able to assess dietary quality of Chinese school-age children, was sensitive to demographics, and more comprehensive and accurate than the CHEI.

Tunable Lattice Reconstruction, Triangular Network of Chiral One-Dimensional States, and Bandwidth of Flat Bands in Magic Angle Twisted Bilayer Graphene.

The interplay between interlayer van der Waals interaction and intralayer lattice distortion can lead to structural reconstruction in slightly twisted bilayer graphene (TBG) with the twist angle being smaller than a characteristic angle θ_{c}. Experimentally, the θ_{c} is demonstrated to be very close to the magic angle (θ≈1.08°). Here we address the transition between reconstructed and unreconstructed structures of the TBG across the magic angle by using scanning tunneling microscopy (STM). Our experiment demonstrates that both structures are stable in the TBG around the magic angle. By using a STM tip, we show that the two structures can be changed to each other and a triangular network of chiral one-dimensional states hosted by domain boundaries can be switched on and off. Consequently, the bandwidth of the flat band, which plays a vital role in the emergent strongly correlated states in the magic angle TBG, is tuned. This provides an extra control knob to manipulate the exotic electronic states of the TBG near the magic angle.

Health information needs regarding diabetes mellitus in China: an internet-based analysis.

BACKGROUND: Today,. most people use the Internet to seek online health-related information from general public health-related websites and discussion groups. However, there are no Internet-based analyses of health information needs pertaining to diabetes in China until now. With the development of artificial intelligence,we can analyzed these online health-related information and provide references for health providers to improve their health service. METHODS: We have done a study of statistically analyzing the questions about diabetes collected from 39 health website, the number of which is 151,589. We have divided these questions into 9 categories using a convolutional neural network. RESULTS: The diabetes problems of consumer are presented as follows, diagnosis: 34.95%, treatment: 25.17%, lifestyle: 21.09%, complication: 8.00%, maternity-related:5.00%, prognosis: 2.59%, health provider choosing: 1.40%, prevention: 1.23%, others: 0.58%, The elderly are more concerned about the treatment and complications of diabetes, while the young are more concerned about the maternity-related and prognosis of diabetes. The diabetes drugs most frequently mentioned by consumers are insulin, metformin and Xiaoke pills, The most concerned complication is caidiovascular disease and diabetic eye disease. CONCLUSION: Diabetes health education should focus on how to prevent diabetes and the contents of health education should be different for differernt age groups;on diabetes treatment, the use of insulin and oral hypoglycemic drugs education should be strengthened.

A novel messenger RNA and long noncoding RNA signature associated with the progression of nonmuscle invasive bladder cancer.

AIM: To explore the molecular mechanism of nonmuscle invasive bladder cancer (NMIBC), matched normal, and cancer tissues of 10 NMIBC were examined for RNA sequencing. METHODS: We profiled the messenger RNA (mRNA) and long noncoding RNA (lncRNA) expression of patients with NMIBC. Differentially expressed mRNAs and lncRNAs were screened between cancer and normal tissues and validated by quantitative polymerase chain reaction (qPCR), and lncRNA-mRNA-miRNA interaction network was constructed. RESULTS: A total of 91 upregulated and 190 downregulated genes and 34 upregulated and 58 downregulated lncRNAs were screened from the sequencing result. The differentially expressed mRNAs were enriched in focal adhesion, rap1 signaling pathway, Hippo signaling pathway, PI3K-Akt signaling pathway, extracellular matrix (ECM)-receptor interaction, Ras signaling pathway, and mitogen-activated protein kinases signaling pathway, of which some pathways were involved in the cancer development. In the RNA sequencing, KIT and laminin subunitγ γ3 (LAMC3) were significantly downregulated in the NMIBC group compared with the normal group. The results of quantitative reverse transcription PCR showed that the expression of LAMC3 and KIT were significantly decreased in the NMIBC group compared with the normal group. The lncRNA-mRNA-miRNA interaction network was constructed by Cytoscape software to further investigate the interaction correlations. The results implied that KIT and LAMC3 might regulate the lncRNAs (such as ENST00000445707, ENST00000501122, ENST00000505254, ENST00000528986, ENST00000557661, ENST00000602964, ENST00000614517, ENST00000620864, and ENST00000623414) by the miRNAs (such as hsa-let-7f-2-3p, hsa-miR-125a-3p, hsa-miR-134-3p, hsa-miR-191-5p, hsa-miR-210-5p, hsa-miR-30a-5p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-92a-2-5p, and hsa-miR-95-3p), and finally played a role in the development of NMIBC cancer. CONCLUSION: Altogether, our study preliminarily indicated that KIT and LAMC3 might play a crucial role in the development of NMIBC cancer via a complex mRNA-lncRNA-miRNA regulatory network.

[Evaluation and Comparison of SHA.LIN,S.T.O.N.E.Nephrolithometry Scoring System,and Clinical Research Office of the Endourological Society Nephrolithometry Nomogram for Predicting Stone Free Rate and Postoperative Outcomes after Percutaneous Nephrolithotomy].

To compare the accuracy of SHA.LIN,S.T.O.N.E.nephrolithometry scoring system,and Clinical Research Office of the Endourological Society(CROES)nephrolithometry nomogram in predicting percutaneous nephrolithotomy(PCNL)outcomes including stone free rate(SFR)and perioperative status. Methods The clinical data of 90 patients with nephrolithiasis undergoing PCNL in department of urology,China-Japan Friendship Hospital from January 2015 to March 2018 were retrospectively analyzed.The general data,stone characteristics,operation approaches,and perioperative variables were recorded.SHA.LIN,S.T.O.N.E.score,and CROES nomogram were assigned according to the computed tomography(CT)findings before surgery.Stone free status was evaluated by kidney-ureter-bladder one month after PCNL.The relationships of SHA.LIN score,S.T.O.N.E.score,and CROES score with SFR,postoperative complications,operation time(OT),length of hospital stay(LOS),estimated blood loss(EBL),and decrease of hemoglobin was evaluated.Receiver operating characteristic(ROC)curves were used to analyze the predictive accuracy. Results The SFR was 72.2%(65/90)and postoperative complications occurred in 33 cases(36.7%).The mean OT was(103.1±39.6)min,the mean EBL was(46.1±53.0)ml,the mean LOS was(15.3±5.2)d,the mean postoperative LOS was(8.5±3.4)d,and the mean decrease of hemoglobin was(16.1±10.2)g/L.Stone-free patients had significantly lower SHA.LIN score(8.23 vs. 10.36,P=0.000)and S.T.O.N.E.score(7.05 vs.8.16,P=0.000)and significantly higher CROES score(188.50 vs. 143.89,P=0.000)compared to patients with residual fragments.All these scores were not significantly associated with complications(P>0.05).On the other hand,all these scores were significantly correlated with OT,EBL,and decrease of hemoglobin(SHA.LIN:POT=0.006,PEBL=0.028,Pdecrease of hemoglobin=0.014;S.T.O.N.E.:POT=0.012,PEBL=0.047,Pdecrease of hemoglobin=0.011;and CROES:POT=0.040,PEBL=0.045,Pdecrease of hemoglobin=0.013).SHA.LIN(P=0.001)and S.T.O.N.E.(P=0.005)scores were associated with LOS.Logistic regression analysis revealed that SHA.LIN(OR=2.491),S.T.O.N.E.(OR=3.030),and CROES(OR=0.973)scores were significantly associated with stone-free status.ROC curves in predicting SFR showed that there was significant difference in the areas under the curves(AUC)for the SHA.LIN vs. S.T.O.N.E.score [0.808(95% CI=0.711-0.905)vs. 0.748(95% CI=0.632-0.864),P=0.047].AUC for the CROES score [0.770(95% CI=0.664-0.877)] showed no significantly different for the SHA.LIN score or the S.T.O.N.E.score(P>0.05). Conclusions All these three scoring systems have good predictive accuracy for SFR.SHA.LIN is more precise than S.T.O.N.E.in predicting SFR.However,they can not predict postoperative complications.

Solitary Fibrous Tumor of the Kidney Treated with Laparoscopic Partial Nephrectomy: A Case Report.

We herein reported a 27-year-old woman with a right renal mass for two years. She underwent laparoscopic partial nephrectomy. Immunohistochemical examination of the specimen confirmed the diagnosis of solitary fibrous tumor by revealing its positive staining for cluster of differentiation (CD)34, epithelial membrane antigen (EMA), B-cell lymphoma-2 (Bcl-2) and CD99 in the tumor cells. No adjuvant treatment was carried out. The patient was in good health without local recurrence or metastasis during 2 years of follow-up. Laparoscopic partial nephrectomy for renal solitary fibrous tumor is an alternative treatment to radical nephrectomy. It can provide a good outcome. However, further follow-up and more cases of renal solitary fibrous tumor treated with laparoscopic partial nephrectomy are necessary to compare the oncological outcome with radical nephrectomy.

SUMO E3 Ligases GmSIZ1a and GmSIZ1b regulate vegetative growth in soybean .

SIZ1 is a small ubiquitin-related modifier (SUMO) E3 ligase that mediates post-translational SUMO modification of target proteins and thereby regulates developmental processes and hormonal and environmental stress responses in Arabidopsis. However, the role of SUMO E3 ligases in crop plants is largely unknown. Here, we identified and characterized two Glycine max (soybean) SUMO E3 ligases, GmSIZ1a and GmSIZ1b. Expression of GmSIZ1a and GmSIZ1b was induced in response to salicylic acid (SA), heat, and dehydration treatment, but not in response to cold, abscisic acid (ABA), and NaCl treatment. Although GmSIZ1a was expressed at higher levels than GmSIZ1b, both genes encoded proteins with SUMO E3 ligase activity in vivo. Heterologous expression of GmSIZ1a or GmSIZ1b rescued the mutant phenotype of Arabidopsis siz1-2, including dwarfism, constitutively activated expression of pathogen-related genes, and ABA-sensitive seed germination. Simultaneous downregulation of GmSIZ1a and GmSIZ1b (GmSIZ1a/b) using RNA interference (RNAi)-mediated gene silencing decreased heat shock-induced SUMO conjugation in soybean. Moreover, GmSIZ1RNAi plants exhibited reduced plant height and leaf size. However, unlike Arabidopsis siz1-2 mutant plants, flowering time and SA levels were not significantly altered in GmSIZ1RNAi plants. Taken together, our results indicate that GmSIZ1a and GmSIZ1b mediate SUMO modification and positively regulate vegetative growth in soybean.

CYCLIN H;1 regulates drought stress responses and blue light-induced stomatal opening by inhibiting reactive oxygen species accumulation in Arabidopsis.

Arabidopsis (Arabidopsis thaliana) CYCLIN-DEPENDENT KINASE Ds (CDKDs) phosphorylate the C-terminal domain of the largest subunit of RNA polymerase II. Arabidopsis CYCLIN H;1 (CYCH;1) interacts with and activates CDKDs; however, the physiological function of CYCH;1 has not been determined. Here, we report that CYCH;1, which is localized to the nucleus, positively regulates blue light-induced stomatal opening. Reduced-function cych;1 RNA interference (cych;1 RNAi) plants exhibited a drought tolerance phenotype. CYCH;1 is predominantly expressed in guard cells, and its expression was substantially down-regulated by dehydration. Transpiration of intact leaves was reduced in cych;1 RNAi plants compared with the wild-type control in light but not in darkness. CYCH;1 down-regulation impaired blue light-induced stomatal opening but did not affect guard cell development or abscisic acid-mediated stomatal closure. Microarray and real-time polymerase chain reaction analyses indicated that CYCH;1 did not regulate the expression of abscisic acid-responsive genes or light-induced stomatal opening signaling determinants, such as MYB60, MYB61, Hypersensitive to red and blue1, and Protein phosphatase7. CYCH;1 down-regulation induced the expression of redox homeostasis genes, such as LIPOXYGENASE3 (LOX3), LOX4, ARABIDOPSIS GLUTATHIONE PEROXIDASE 7 (ATGPX7), EARLY LIGHT-INDUCIBLE PROTEIN1 (ELIP1), and ELIP2, and increased hydrogen peroxide production in guard cells. Furthermore, loss-of-function mutations in CDKD;2 or CDKD;3 did not affect responsiveness to drought stress, suggesting that CYCH;1 regulates the drought stress response in a CDKD-independent manner. We propose that CYCH;1 regulates blue light-mediated stomatal opening by controlling reactive oxygen species homeostasis.

Visualizing a single wavefront dislocation induced by orbital angular momentum in graphene.

Phase singularities are phase-indeterminate points where wave amplitudes are zero, which manifest as phase vertices or wavefront dislocations. In the realm of optical and electron beams, the phase singularity has been extensively explored, demonstrating a profound connection to orbital angular momentum. Direct local imaging of the impact of orbital angular momentum on phase singularities at the nanoscale, however, remains challenging. Here, we study the role of orbital angular momentum in phase singularities in graphene, particularly at the atomic level, through scanning tunneling microscopy and spectroscopy. Our experiments demonstrate that the scatterings between different orbital angular momentum states, which are induced by local rotational symmetry-breaking potentials, can generate additional phase singularities, and result in robust single-wavefront dislocations in real space. Our results pave the way for exploring the effects of orbital degree of freedom on quantum phases in quasiparticle interference processes.

Design and application of specific 16S rDNA-targeted primers for assessing endophytic diversity in Dendrobium officinale using nested PCR-DGGE.

Novel specific 16S rDNA-targeted primers were successfully designed and applied to the characterization of endophytic diversity in Dendrobium officinale. Using the popular universal bacterial primers 27f/1492r, the fragments of chloroplast and mitochondrion 16S/18S rDNA were amplified from D. officinale. They shared high nucleotide identity with the chloroplast 16S rDNAs (99-100 %) and with the mitochondrion 18S rDNAs (93-100 %) from various plants, respectively, and both shared 73-86 % identities with the bacterial 16S rDNA sequences in GenBank. The current bacterial universal primers, including 27f/1492r, match well with the chloroplast and mitochondrion 16S/18S rDNAs, which accordingly renders these primers not useful for endophytic diversity analysis. Novel 16S rDNA-targeted primers fM1 (5'-CCGCGTGNRBGAHGAAGGYYYT-3') and rC5 (5'-TAATCCTGTTTGCTCC CCAC-3') were designed, which show good specificity compared to the 16S/18S rDNAs of D. officinale, and perfect universality within bacteria except for Cyanobacteria. The primers fM1/rC5, together with 515f-GC/rC5, which overlaps the whole V4 region of 16S rDNA, were subjected to nested polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) to analyze the diversity of endophytic bacteria in D. officinale from three different sources in China. The results showed diversities in roots and stems of the plants from all three locations. Altogether, 29 bands were identified as bacteria, with the dominant group being Proteobacteria and the dominant genus being Burkholderia, some of which commonly has the function of nitrogen fixation and thus may play potentially important roles in D. officinale. Therefore, the nested PCR-DGGE method based on the novel primers provides a good alternative for investigating the communities and roles of endophytes in D. officinale.

Nitrogen nutrition diagnosis and estimation of seed cotton yield based on critical nitrogen concentration for drip irrigation cotton in Southern Xinjiang, China.

The stability and adaptability of the critical nitrogen concentration in drip-irrigated cotton fields in Southern Xinjiang were investigated by measuring the nitrogen uptake (Nuptake), nitrogen nutrition index (NNI), and accumulated nitrogen deficit (Nand). A two-year field study was conducted using five nitrogen levels (0, 75, 150, 300, and 450 kg·hm-2), with cotton cultivars 'Xinluzhong55' and 'Xinluzhong78' as the test materials. We analyzed the effects of nitrogen addition on plant dry matter (PDM) and critical nitrogen concentration dilution curves (PNCc). The Nuptake, NNI, and Nand models as well as a model of the relationships between NNI, Nand, and the relative seed cotton yield (RY) were established based on the PNCc for both cultivars. The results showed that the PNCc, maximum nitrogen concentration (PNCmax), and minimum nitrogen concentration (PNCmin) dilution models in drip-irrigated cotton showed power function relationships with plant dry matter (PDM), with determination coefficients (R2) of 0.905 and 0.960, 0.919 and 0.950, 0.934 and 0.969 for Xinluzhong55 and Xinluzhong78, respectively. Model validation with independent experimental data showed that the critical nitrogen concentrations in cotton grown under drip-irrigated conditions were simulated satisfactorily. The Xinluzhong55 and Xinluzhong78 1:1 line R2, root mean square error (RMSE) and relative estimation error (RE) of the simulated and observed critical nitrogen concentrations were 0.987, 0.051 g·kg-1, 1.95% and 0.984, 0.044 g·kg-1, 1.91%, respectively, showing good stability and adaptability between the two years. According to the Nuptake, NNI, and Nand model, we concluded that 300 kg·hm-2 was the optimum level of nitrogen application in drip-irrigated cotton in Southern Xinjiang. Based on the fitting results of nitrogen level and seed cotton yield, the theoretical appropriate nitrogen applications for Xinluzhong55 and Xinluzhong78 were 322.7 and 336.4 kg·hm-2, respectively. The R2 of NNI, Nand and RY in different growth stages were greater than 0.900, the NNI, Nand and RY estimation model of seed cotton yield based on 1:1 lines of R2, RMSE, and RE were 0.899-0.989, 0.05-0.13, and 4.1%-9.9% and 0.902-0.981, 0.04-0.12, and 5.1%-9.5%, all of which were highly significant. These results could provide new directions for the evaluation and diagnosis of optimum nitrogen application levels and nitrogen status, as well as the estimation of seed cotton yield in drip-irrigated cotton in Southern Xinjiang.

Dynamic alteration and prognostic significance of tumor-associated CD68+ and CD68+ PD-L1- macrophages in muscle-invasive bladder cancer treated with neoadjuvant chemotherapy.

BACKGROUND: The current study aimed to investigate the dynamic alteration and prognostic significance of tumor-infiltrating lymphocytes (TILs), tumor-associated macrophages (TAMs), and PD-L1 status of immune cells in muscle-invasive bladder cancer (MIBC) treated with neoadjuvant chemotherapy (NAC). METHODS: Multiplex immunofluorescence staining was performed to examine CD68+ TAM, CD4+ T cell, CD8+ T cell, FOXP3+ Treg cell, and PD-L1 expression in paired MIBC tissues (n = 54) before and after NAC. Patients were then divided into definite responders (DR), (≤pT1) and incomplete responders (IR). RESULTS: There was no significant difference between DR and IR cohorts for the immune cell infiltration levels at the baseline status. Tobacco history was identified to be associated with worse NAC efficacy. CD68+ (stroma area: p = 0.025; tumor area: p = 0.028; total area: p = 0.013) and CD68+ PD-L1- (stroma area: p = 0.035; tumor area: p = 0.013 total area: p = 0.014) TAMs infiltration levels decreased significantly after NAC, while there was no significant difference of CD68+ PD-L1+ and TILs. The infiltration of CD68+ (p = 0.033), CD68+ PD-L1- (p = 0.033), and CD68+ PD-L1+ (p < 0.001) TAMs in stroma area were significantly associated with poorer disease-free survival rate (DFS) of MIBC patients. CONCLUSION: CD68+ and CD68+ PD-L1- TAMs infiltration levels decreased significantly after NAC and pre-treatment TAM infiltration levels were independent prognostic factors for MIBC patients. While there was no sufficient evidence demonstrating that pre-treatment TILs or TAMs could predict response to NAC in MIBC patients.