[Incidence and related factors of chronic neuropathic pain in elderly patients after video-assisted thoracoscopic surgery].

Objective: To investigate the incidence and related factors of chronic neuropathic pain (CNP) in elderly patients after thoracoscopic surgery. Methods: A total of 463 elderly patients (aged≥60 years) who underwent elective video-assisted thoracoscopic surgery from November 2020 to May 2021 at Peking Union Medical College Hospital were prospectively recruited. Among them, 283 were males and 180 were females, with an average age of (66.6±4.8) years. Chronic postsurgical pain (CPSP) was assessed by telephone at 6 months after surgery, and then the patients with CNP were screened using the ID-pain scale. Multivariable logistic regression was used to analyze the related factors for CNP in elderly patients after thoracoscopic surgery. Results: The incidence of CPSP was 41.9% (194/463), and the incidence of CNP was 18.8% (87/463). Multivariable logistic regression analysis showed that incision number<3 (OR=0.385, 95%CI: 0.156-0.949, P=0.038) and intraoperative N2O inhalation (OR=0.506, 95%CI: 0.304-0.842, P=0.009) were protective factors for CNP in elderly patients after thoracoscopic surgery, but high numeric rating scale (NRS) score on the first day after surgery (OR=1.180, 95%CI: 1.056-1.318, P=0.003) was a risk factor. Conclusions: The incidence of CNP in elderly patients after thoracoscopic surgery is 18.8%. Incision number<3 and intraoperative N2O inhalation are protective factors for CNP, but high NRS score on the first day after surgery is a risk factor.

LncRNA MIR503HG regulated cell viability, metastasis and apoptosis of cervical cancer via miR-191/CEBPB axis.

OBJECTIVE: The long non-coding RNA MIR503 host gene (MIR503HG) plays a role in suppressing or promoting cancer in many types of human malignant tumors. The role of MIR503HG in cervical cancer is still unknown. PATIENTS AND METHODS: The expression level of MIR503HG in cervical cancer tissues and cell lines was accessed using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay. The Cell Counting Kit-8 (CCK-8) assay and flow cytometric analysis were performed to assess cell proliferation and apoptosis in cervical cancer. The nude mouse xenograft experiment was used to examine the ability of MIR503HG in tumor formation. In our study, we found that the expression of MIR503HG was significantly reduced in cervical cancer tissues and cell lines. In vitro studies have shown that MIR503HG inhibited cell proliferation and invasion, and enhanced cell apoptosis in cervical cancer through the miR-191/CEBPB axis. MIR503HG regulated the expression of miR-191 via directly binding to miR-191. RESULTS: The expression of MIR503HG had a negative correlation with miR-191 expression in cervical cancer tissues. MiR-191 regulated the expression of CEBPB by directly targeting 3'-UTR of CEBPB mRNA. Overexpression of MIR503HG inhibited cell proliferation, invasion and apoptosis in vitro, and inhibited tumor growth in vivo. CONCLUSIONS: MIR503HG plays a role in suppressing tumors in cervical cancer and is a long-term non-coding RNA.

[Comparison of efficacy between sorafenib and sunitinib as first-line therapy for metastatic renal cell carcinoma and analyze prognostic factors for survival].

Objective: To investigate the efficacy and drug related adverse reactions of sorafenib and sunitinib as first-line tyrosine-kinase inhibitors (TKIs) for patients with metastatic renal cell carcinoma (mRCC) and analyze the clinical prognostic factor for survival. Methods: The data of 271 patients with metastatic renal cell carcinoma who had complete clinicopathological data were retrospectively analyzed, including 174 cases in sorafenib group and 97 cases in sunitinib group, to access patients' overall survival (OS) and progression-free survival (PFS). Prognostic values of all characteristics were determined by using univariate and multivariate Cox regression models. Results: The objective response rates (ORR) of the sorafenib and sunitinib groups were 14.9% and 19.6%, respectively, and the disease control rates (DCR) were 85.1% and 88.6%, respectively. No significant difference was found between the sorafenib and sunitinib group in ORR (P=0.325) or DCR (P=0.408). The most common grade 3 to 4 adverse events in the sorafenib group were hand-foot syndrome (6.7%), diarrhea (2.3%), and rash (2.3%). The most common grade 3 to 4 adverse events in the sunitinib group were neutropenia (6.2%), hand-foot syndrome (6.2%), and thrombocytopenia (4.6%). During the follow-up, 97 cases death occurred and 81 cases disease progression occurred in sorafenib group. The median PFS was 12 months (95% CI: 9-15 months), and the median OS was 25 months (95% CI: 21-29 months) in sorafenib group. While 74 cases death occurred and 40 cases disease progression occurred in sunitinib group, the median PFS was 12 months (95% CI: 10-12 months) and the median OS was 23 months (95% CI: 20-32 months) in sunitinib group. No significant difference was found between the sorafenib and the sunitinib group in PFS (P=0.771) or OS (P=0.548). Multivariate analysis showed Fuhrman grades (HR=1.358, 95%CI: 1.004-1.835), number of metastatic sites (HR=1.550, 95%CI: 1.143-2.101) and MSKCC risk grade (Intermediate risk group: HR=1.621, 95%CI: 1.117-2.232; Poor risk group: HR=2.890, 95%CI: 1.942-4.298) were independent prognostic factors for PFS. Fuhrman grades (HR=2.135, 95%CI: 1.533-2.974), number of metastatic sites (HR=1.774, 95%CI: 1.279-2.461) and MSKCC risk grade (Intermediate risk group: HR=1.415, 95%CI: 1.002-1.998; Poor risk group: HR=3.161, 95%CI: 2.065-4.838) were independent prognostic factors for OS. Conclusions: The results of this study indicate that sorafenib and sunitinib are both effective as the first-line TKIs for mRCC patients and sorafenib has comparable efficacy to sunitinib. But they have differences in the incidence of adverse effects. Fuhrman grades, number of metastatic sites and MSKCC risk grade are independent prognostic factors for mRCC patients.

Genetic diversity and genetic structure of consecutive breeding generations of golden mandarin fish (Siniperca scherzeri Steindachner) using microsatellite markers.

In this study, 12 polymorphic microsatellites were inves-tigated to determine the genetic diversity and structure of 5 consecu-tive selected populations of golden mandarin fish (Siniperca scherzeri Steindachner). The total numbers of alleles, average heterozyosity, and average polymorphism information content showed that the genetic diversity of these breeding populations was decreasing. Additionally, pairwise fixation index FST values among populations and Da values in-creased from F1 generation to subsequent generations (FST values from 0.0221-0.1408; Da values from 0.0608-0.1951). Analysis of molecular variance indicated that most genetic variations arise from individuals within populations (about 92.05%), while variation among populations accounted for only 7.95%. The allele frequency of the loci SC75-220 and SC101-222 bp changed regularly in the 5 breeding generations. Their frequencies were gradually increased and showed an enrichment trend, indicating that there may be genetic correlations between these 2 loci and breeding traits. Our study indicated that microsatellite markers are effective for assessing the genetic variability in the golden mandarin fish breeding program.

Widespread exon skipping triggers degradation by nuclear RNA surveillance in fission yeast.

Exon skipping is considered a principal mechanism by which eukaryotic cells expand their transcriptome and proteome repertoires, creating different splice variants with distinct cellular functions. Here we analyze RNA-seq data from 116 transcriptomes in fission yeast (Schizosaccharomyces pombe), covering multiple physiological conditions as well as transcriptional and RNA processing mutants. We applied brute-force algorithms to detect all possible exon-skipping events, which were widespread but rare compared to normal splicing events. Exon-skipping events increased in cells deficient for the nuclear exosome or the 5'-3' exonuclease Dhp1, and also at late stages of meiotic differentiation when nuclear-exosome transcripts decreased. The pervasive exon-skipping transcripts were stochastic, did not increase in specific physiological conditions, and were mostly present at less than one copy per cell, even in the absence of nuclear RNA surveillance and during late meiosis. These exon-skipping transcripts are therefore unlikely to be functional and may reflect splicing errors that are actively removed by nuclear RNA surveillance. The average splicing rate by exon skipping was ∼ 0.24% in wild type and ∼ 1.75% in nuclear exonuclease mutants. We also detected approximately 250 circular RNAs derived from single or multiple exons. These circular RNAs were rare and stochastic, although a few became stabilized during quiescence and in splicing mutants. Using an exhaustive search algorithm, we also uncovered thousands of previously unknown splice sites, indicating pervasive splicing; yet most of these splicing variants were cryptic and increased in nuclear degradation mutants. This study highlights widespread but low frequency alternative or aberrant splicing events that are targeted by nuclear RNA surveillance.

The FEN1 E359K germline mutation disrupts the FEN1-WRN interaction and FEN1 GEN activity, causing aneuploidy-associated cancers.

Polymorphisms and somatic mutations in Flap Endonuclease 1 (FEN1), an essential enzyme involved in DNA replication and repair, can lead to functional deficiencies of the FEN1 protein and a predisposition to cancer. We identified a FEN1 germline mutation that changed residue E359 to K in a patient whose family had a history of breast cancer. We determined that the E359K mutation, which is in the protein-protein domain of FEN1, abolished the interaction of FEN1 with Werner syndrome protein (WRN), an interaction that is critical for resolving stalled DNA replication forks. Furthermore, although the flap endonuclease activity of FEN1 E359K was unaffected, it failed to resolve bubble structures, which require the FEN1 gap-dependent endonuclease activity. To determine the etiological significance of E359K, we established a mouse model containing this mutation. E359K mouse embryonic fibroblasts (MEF) were more sensitive to DNA crosslinking agents that cause replication forks to stall. Cytological analysis suggested that the FEN1-WRN interaction was also required for telomere stability; mutant cell lines had fragile telomeres, increased numbers of spontaneous chromosomal anomalies and higher frequencies of transformation. Moreover, the incidence of cancer was significantly higher in mice homozygous for FEN1 E359K than in wild-type mice, suggesting that the FEN1 E359K mutation is oncogenic.

LaSSO, a strategy for genome-wide mapping of intronic lariats and branch points using RNA-seq.

Both canonical and alternative splicing of RNAs are governed by intronic sequence elements and produce transient lariat structures fastened by branch points within introns. To map precisely the location of branch points on a genomic scale, we developed LaSSO (Lariat Sequence Site Origin), a data-driven algorithm which utilizes RNA-seq data. Using fission yeast cells lacking the debranching enzyme Dbr1, LaSSO not only accurately identified canonical splicing events, but also pinpointed novel, but rare, exon-skipping events, which may reflect aberrantly spliced transcripts. Compromised intron turnover perturbed gene regulation at multiple levels, including splicing and protein translation. Notably, Dbr1 function was also critical for the expression of mitochondrial genes and for the processing of self-spliced mitochondrial introns. LaSSO showed better sensitivity and accuracy than algorithms used for computational branch-point prediction or for empirical branch-point determination. Even when applied to a human data set acquired in the presence of debranching activity, LaSSO identified both canonical and exon-skipping branch points. LaSSO thus provides an effective approach for defining high-resolution maps of branch-site sequences and intronic elements on a genomic scale. LaSSO should be useful to validate introns and uncover branch-point sequences in any eukaryote, and it could be integrated into RNA-seq pipelines.

Development and characterization of novel SSR markers in Siniperca kneri Garman.

In this study, 37 transcriptome-derived simple sequence repeat (SSR) markers and 18 genomic SSR markers were developed and characterized in the Chinese perch, Siniperca kneri Garman. The average allele number per locus was 5.1 (range: 2-8) for transcriptome-derived SSRs and 3.8 (range: 2-5) for genomic SSRs. The average observed and expected heterozygosities were 0.666 (range: 0.000-1.000) and 0.692 (range: 0.230-0.857) for transcriptome-derived SSRs, respectively. These values were 0.380 (range: 0.000-1.000) and 0.527 (range: 0.201-0.799) for genomic SSRs, respectively. The average polymorphic information content was 0.638 (range: 0.215-0.824) for transcriptome-derived SSRs and 0.477 (range: 0.183-0.752) for genomic SSRs. Seven of these loci exhibited departure from Hardy-Weinberg equilibrium after sequential Bonferroni's correction for multiple tests, and no significant deviation was observed for the linkage disequilibrium. These developed and characterized markers are anticipated to be useful for studies on population genetics, conservation genetics, and the fishery management of this species.

New microsatellite loci for the mandarin fish Siniperca chuatsi and their application in population genetic analysis.

The mandarin fish is a popular fresh water food fish in China. Fifty-three polymorphic microsatellite markers were isolated through construction of an enriched library of genomic DNA of Siniperca chuatsi (Percichthyidae). We found 2 to 7 alleles per locus. The observed and expected heterozygosity values varied from 0.059 to 1.000 and from 0.305 to 0.818, respectively. The polymorphic information content value varied from 0.255 to 0.782. Twelve microsatellite loci deviated significantly from Hardy-Weinberg equilibrium after Bonferroni's correction. These markers were evaluated in five species of sinipercine fish; 98% of the 265 locus/taxon combinations tested gave cross-amplification. Eight polymorphic microsatellite markers were randomly selected for genetic characterization of three S. chuatsi populations. The Ganjiang River and Yuanjiang River populations had moderate levels of genetic diversity, while the Mudanjiang River population had a relatively low level genetic diversity. Genetic distance-based cluster analysis showed clustering of the Ganjiang River and Yuanjiang River populations in a single group and the Mudanjiang River population in a separate group. Based on these results, we suggest that S. chuatsi from the Yangtze River watershed are distinct from the Mudanjiang River population. These SSR markers will be useful for diversity, mapping and marker assisted studies of S. chuatsi and other sinipercine fishes.

First Report of a Leaf Spot of Radermachera sinica in China Caused by Bacillus megaterium.

Radermachera sinica is widely planted as an ornamental plant in homes, offices, and malls in China. A leaf spot of R. sinica occurred in Luoyang, China, from 2013 to 2014. Lesions mostly occurred in wounds and were irregular with light brown centers and purple borders. One or more lesions on a leaf sometimes covered the entire blade. Eighty plants were surveyed in Luoyang, with disease incidence of 17%. Five millimeter pieces from the borders of lesions were surface-disinfected with 75% ethanol for 30 s, 1% sodium hypochlorite for 5 min, washed three times in sterilized distilled water, placed on nutrient agar (NA) medium at 25°C in darkness, and incubated for 24 to 48 h. Four white, round, smooth, and shiny colonies were selected for further identification. All strains were gram-positive, aerobic rods with many peritrichous flagella, and could grow in medium containing 5% NaCl. The strains were positive for catalase, starch hydrolysis, liquefaction of gelatin, reduction of nitrate, acid production from glucose, mannitol, maltose, lactose, xylose, and pectinose. The strains were positive for phenylalanine deaminase, decomposition of tyrosine, and utilization of citrate. The strains were identified by biochemical tests as Bacillus megaterium (1). To confirm pathogenicity, the strains were grown on NA for 48 h and suspended in sterile distilled water to produce a suspension with a final concentration of 108 CFU/ml. Healthy leaves of biennial R. sinica plants were sterilized with 75% ethanol and washed three times with sterilized distilled water. Fresh wounds were made with a sterile needle on the healthy leaves. Each of four strains was tested by spray inoculation with a bacterial suspension on three leaves. Sterile distilled water was used as negative control. Plants were enclosed in plastic bags and placed in a growth chamber at 28°C with 80% relative humidity. After 5 days, water-soaked lesions were observed. Two weeks later, lesions 4 mm in diameter turned light brown with purple borders, and most of lesions occurred in puncture wounds. Symptoms similar to those observed on field plants developed on all inoculated leaves, while no symptoms appeared on the control leaves. B. megaterium was re-isolated from the lesions of inoculated leaves, but not from the control leaves. To confirm the bacterial identification, PCR was performed on the 16S rDNA gene with P1/P2 (P1: CAGAGTTTGATCCTGGCT, P2: AGGAGGTGATCCAGCCGCA) (2) and 1,463 bp of the 16S rDNA gene (GenBank Accession No. KJ789369) showed 100% sequence identity to B. megaterium DSM 319 (NC_014103.1). To our knowledge, this is the first report of a leaf spot of R. sinica caused by B. megaterium in China as well as anywhere in the world. References: (1) P. Vos et al. Bergey's Manual of Systematic Bacteriology. Vol 3: The Firmicutes. Springer, 2009. (2) W. G. Weisbury et al. J. Bacteriol. 173:697, 1991.

Nonmuscle myosin IIA is associated with poor prognosis of esophageal squamous cancer.

Nonmuscle myosin IIA (myosin IIA) is a force-producing protein involved in the process of cell migration. Its expression has been considered as a bad prognostic indicator in stage I lung adenocarcinoma. However, the expression and clinical significance of myosin IIA in esophageal cancer has not been explored. In this study, we investigate the expression level of myosin IIA in 50 esophageal squamous cancer and 30 adjacent normal esophageal tissues by immunohistochemical staining and correlated its expression with clinicopathological features. Myosin IIA was expressed in all esophageal squamous cancer tissues (100%) and 8 of 30 adjacent normal tissues (26.7%, P = 0.000). In cancer tissues, elevated myosin IIA expression level was significantly correlated with increasing metastatic lymph nodes, poorer cancer differentiation, and advanced tumor stage. Further univariate analysis suggested that strong myosin IIA expression was associated with a significantly shorter overall survival (P = 0.021). In addition, MYH9 SiRNA was transfected into esophageal squamous cancer cell line (KYSE-510) to study the role of myosin IIA in cell migration. SiRNA-mediated depletion of myosin IIA in KYSE-510 cells significantly increased cell-matrix adhesion and attenuated cell migration ability (P = 0.000). In conclusion, these findings indicate that overexpression of myosin IIA may contribute to the progression and poor prognosis of esophageal squamous cancer, and this effect may be associated with increased cancer cell migration.

Modified thoracoscopic versus minimally invasive oesophagectomy in curative resection of oesophageal cancer.

Conventional thoracoscopic oesophagectomy is time-consuming and requires sophisticated endoscopic skills. To reduce these problems we have modified the operating procedure, first by anastomosis of the oesophagus with the tubular stomach pulled up via the retrosternal route, followed by thoracoscopic oesophagectomy (modified thoracoscopic oesophagectomy). Outcomes were compared between the modified procedure and minimally invasive oesophagectomy. There were no significant differences in general preoperative clinical characteristics between the two patient groups. The modified thoracoscopic oesophagectomy group had significantly lower hospitalization expenses, significantly shorter operation times and significantly more lymph nodes removed compared with the minimally invasive oesophagectomy group, but there were no significant group differences in lengths of hospital and intensive care unit stays, morbidity and mortality. These results indicate that modified thoracoscopic oesophagectomy is feasible, simplifies operating procedures and reduces hospitalization expenses with acceptable morbidity.

FZD7 has a critical role in cell proliferation in triple negative breast cancer.

Breast cancer is genetically and clinically heterogeneous. Triple negative breast cancer (TNBC) is a subtype of breast cancer that is usually associated with poor outcome and lack of benefit from targeted therapy. We used microarray analysis to perform a pathway analysis of TNBC compared with non-triple negative breast cancer (non-TNBC). Overexpression of several Wnt pathway genes, such as frizzled homolog 7 (FZD7), low density lipoprotein receptor-related protein 6 and transcription factor 7 (TCF7) was observed in TNBC, and we directed our focus to the Wnt pathway receptor, FZD7. To validate the function of FZD7, FZD7shRNA was used to knock down FZD7 expression. Notably, reduced cell proliferation and suppressed invasiveness and colony formation were observed in TNBC MDA-MB-231 and BT-20 cells. Study of the possible mechanism indicated that these effects occurred through silencing of the canonical Wnt signaling pathway, as evidenced by loss of nuclear accumulation of ß-catenin and decreased transcriptional activity of TCF7. In vivo studies revealed that FZD7shRNA significantly suppressed tumor formation, through reduced cell proliferation, in mice bearing xenografts without FZD7 expression. Our findings suggest that FZD7-involved canonical Wnt signaling pathway is essential for tumorigenesis of TNBC, and thus, FZD7 shows promise as a biomarker and a potential therapeutic target for TNBC.

MILDOS uranium milling dose assessment code update.

The MILDOS-AREA code was developed to estimate radiological doses and risks from uranium milling activities. The code has been used for demonstrating radiological compliance regarding the U.S. Nuclear Regulatory Commission's licensing requirements for uranium milling activities. The code was recently updated with an enhanced software package to address the following four areas: regulatory changes, in-situ leaching extraction technologies, software user interfaces, and software distribution technologies via the internet. Users can now specify in-situ leaching processes through a Windows object-based Geographic information System interface with incorporated updated regulation methodologies. The code and documentation are freely distributed through the Internet.

Conformational flexibility of a ubiquitin conjugation enzyme (E2).

Ubiquitination plays important roles in a variety of biological processes, such as DNA repair, cell cycle regulation, and p53-dependent processes. Despite intensive studies in ubiquitination, the mechanism of substrate recognition is still not well understood. Each E2 has its own substrate specificity, yet substrate proteins recognized by each E2 are highly diverse. To better understand how E2 proteins confer both substrate specificity and diversity, we have studied conformational flexibility of an E2, UBC9, using nuclear magnetic resonance 15N relaxation and hydrogen-deuterium exchange measurements. Two regions in human UBC9 show higher mobility over a wide range of time scales. Combined with previous biochemical studies, both regions are likely to be important for protein-protein recognition in the ubiquitin pathway. The region near the N-terminus may be important for interactions with the E1-UBL1 conjugate. The region near the C-terminus, which undergoes conformational exchange may be important for substrate binding and catalytic activity. Since E2 enzymes share high homology in primary sequences and three-dimensional structures, the conformational flexibility of UBC9 may represent a general feature of E2 enzymes. This study provides a new perspective for further studies of protein-protein recognition in ubiquitination.

A novel DNA-binding motif shares structural homology to DNA replication and repair nucleases and polymerases.

A novel class of DNA-binding domains has been established from at least sixteen recently identified DNA-binding proteins. The three-dimensional structure of one of these domains, Mrf-2, has been solved using NMR methods. This structure is significantly different from known DNA-binding domain structures. The mechanism of DNA recognition by this motif has been suggested based on conserved residues, surface electrostatic potentials and chemical shift changes. This new DNA-binding motif shares structural homology with T4 RNase H, E. coli endonuclease III and Bacillus subtilis DNA polymerase I. The structural homology suggests a mechanism for substrate recognition by these enzymes.

Diagnostic value of occult fecal blood testing for colorectal cancer screening.

AIM: To evaluate the diagnostic value of occult fecal blood testing in mass colorectal cancer screening. METHODS: A reverse passive hemagglutination reaction fecal occult blood test (RPHA-FOBT) and colorectal cancer risk factor quantitative method were used as preliminary screening for colorectal cancer. A 60-cm fiber optic colonoscopy was used to validate the preliminary screen and was used to detect colorectal cancer in a community of 75813 subjects. RESULTS: Compared to the 60-cm fiber optic colonoscopy as a standard reference, FOBT has a sensitivity of 41.9%, specificity of 95.8%, Youden's index of 0.38, and positive predictive value of 0.68%. These results increased with subject age from the first detection. A 3-year follow up in the target mass showed that all new cases had initially been FOBT-negative. CONCLUSION: The value of FOBT as an indicator of colorectal cancer in mass screening is limited.