Sequence comparison of the mitochondrial genomes of Plesionika species (Caridea: Pandalidae), gene rearrangement and phylogenetic relationships of Caridea.

Background: Despite the Caridean shrimps' vast species richness and ecological diversity, controversies persist in their molecular classification. Within Caridea, the Pandalidae family exemplifies significant taxonomic diversity. As of June 25, 2023, GenBank hosts only nine complete mitochondrial genomes (mitogenomes) for this family. The Plesionika genus within Pandalidae is recognized as polyphyletic. To improve our understanding of the mitogenome evolution and phylogenetic relationships of Caridea, this study introduces three novel mitogenome sequences from the Plesionika genus: P.  ortmanni, P. izumiae and P. lophotes. Methods: The complete mitochondrial genomes of three Plesionika species were sequenced utilizing Illumina's next-generation sequencing (NGS) technology. After assembling and annotating the mitogenomes, we conducted structural analyses to examine circular maps, sequence structure characteristics, base composition, amino acid content, and synonymous codon usage frequency. Additionally, phylogenetic analysis was performed by integrating existing mitogenome sequences of true shrimp available in GenBank. Results: The complete mitogenomes of the three Plesionika species encompass 37 canonical genes, comprising 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), two ribosomal RNAs (rRNAs), and one control region (CR). The lengths of these mitogenomes are as follows: 15,908 bp for P. ortmanni, 16,074 bp for P. izumiae and 15,933 bp for P. lophotes. Our analyses extended to their genomic features and structural functions, detailing base composition, gene arrangement, and codon usage. Additionally, we performed selection pressure analysis on the PCGs of all Pandalidae species available in Genbank, indicating evolutionary purification selection acted on the PCGs across Pandalidae species. Compared with the ancestral Caridea, translocation of two tRNA genes, i.e., trnP or trnT, were found in the two newly sequenced Plesionika species-P. izumiae and P. lophotes. We constructed a phylogenetic tree of Caridea using the sequences of 13 PCGs in mitogenomes. The results revealed that family Pandalidae exhibited robust monophyly, while genus Plesionika appeared to be a polyphyletic group. Conclusions: Gene rearrangements within the Pandalidae family were observed for the first time. Furthermore, a significant correlation was discovered between phylogenetics of the Caridea clade and arrangement of mitochondrial genes. Our findings offer a detailed exploration of Plesionika mitogenomes, laying a crucial groundwork for subsequent investigations into genetic diversity, phylogenetic evolution, and selective breeding within this genus.

Influence of trimetazidine on myocardial injury in mice with diabetic cardiomyopathy.

INTRODUCTION: The prevalence of diabetes mellitus is increasing year by year globally, and diabetic cardiomyopathy (DCM), as the most common complication of type 2 diabetes mellitus, seriously affects the prognosis of patients. Trimetazidine (TMZ), as a drug affecting myocardial energy metabolism, mainly reduces the oxidation rate of ß-oxidation by inhibiting 3-ketoacyl-CoA thiolase (3-KAT), a key enzyme in ß-oxidation of free fatty acid (FFA), so that the energy metabolism substrate of cardiomyocytes preferentially selects glucose rather than fatty acids, increases the content of intracellular adenosine triphosphate (ATP), enhances the contractile function of cardiomyocytes, and improves the state of cellular ischemia and hypoxia. Previous studies have shown that TMZ is closely related to the activation and induction of apoptosis of the MAPK pathway and AMPK pathway, and plays a role in the treatment of diabetic cardiomyopathy, but the specific mechanism is still unclear. OBJECTIVE: This study aims to investigate the impact of TMZ on myocardial damage in mice exhibiting diabetic cardiomyopathy (DCM), and to furnish a laboratory foundation for the clinical treatment of diabetic cardiomyopathy. METHOD: Male db/db mice (6 weeks old, n = 21) and male wild-type (wt) (6 weeks old, n = 20) mice were selected for the study. The wt mice were randomly assigned to the wt group (n = 10) and wt + TMZ group (n = 10), while the remaining db/db mice were randomly allocated to the db/db group (n = 11) and db/db + TMZ group (n = 10). Following 8 weeks of feeding, the wt + TMZ group and db/db + TMZ group received TMZ via gavage, whereas the remaining groups were administered physiological saline. Periodic measurements of blood glucose, blood lipids, and myocardial enzymes were conducted in mice, with samples obtained after the 12th week for subsequent biochemical analysis, myocardial pathology assessment, immunohistochemistry, western blot analysis, and TUNEL staining (TdT-mediated dUTP Nick-End Labeling). RESULT: GLU, TC, TG, LDL-C, and CK-MB levels were significantly higher in db/db mice compared to wt mice (GLU: M ± SD wt 5.94 ± 0.37, db/db 17.63 ± 0.89, p < 0.05, ES = 0.991; TC: M ± SD wt 3.01 ± 0.32, db/db 6.97 ± 0.36, p < 0.05, ES = 0.972; TG: M ± SD wt 0.58 ± 0.2, db/db 1.75 ± 0.14, p < 0.05, ES = 0.920; LDL-C: M ± SD wt 1.59 ± 0.12, db/db 3.87 ± 0.14, p < 0.05, ES = 0.989; CK-MB: M ± SD wt 0.12 ± 0.01, db/db 0.31 ± 0.04, p < 0.05, ES = 0.928). HDL-C levels were significantly lower in db/db mice (M ± SD wt 1.89 ± 0.08, db/db 0.64 ± 0.09, p < 0.05, ES = 0.963). Histopathological analysis confirmed myocardial damage in db/db mice. Treatment with TMZ reduced GLU, TC, TG, LDL-C, and CK-MB levels (p < 0.05, ES > 0.9) and increased HDL-C levels compared to untreated db/db mice. Additionally, TMZ treatment significantly decreased myocardial cell apoptosis (p < 0.05, ES = 0.980). These results demonstrate the efficacy of TMZ in reversing myocardial injury in DCM mice. CONCLUSION: TMZ can mitigate myocardial damage in db/db mice by downregulating the expression of caspase-12, a protein associated with the endoplasmic reticulum stress (ERS) cell apoptosis pathway, consequently diminishing cell apoptosis. This underscores the protective efficacy of TMZ against myocardial damage in mice afflicted with DCM.

Characterization of C30 carotenoid and identification of its biosynthetic gene cluster in Methylobacterium extorquens AM1.

Methylobacterium species, the representative bacteria distributed in phyllosphere region of plants, often synthesize carotenoids to resist harmful UV radiations. Methylobacterium extorquens is known to produce a carotenoid pigment and recent research revealed that this carotenoid has a C30 backbone. However, its exact structure remains unknown. In the present study, the carotenoid produced by M. extorquens AM1 was isolated and its structure was determined as 4-[2-O-11Z-octadecenoyl-ß-glucopyranosyl]-4,4'-diapolycopenedioc acid (1), a glycosylated C30 carotenoid. Furthermore, the genes related to the C30 carotenoid synthesis were investigated. Squalene, the precursor of the C30 carotenoid, is synthesized by the co-occurrence of META1p1815, META1p1816 and META1p1817. Further overexpression of the genes related to squalene synthesis improved the titer of carotenoid 1. By using gene deletion and gene complementation experiments, the glycosyltransferase META1p3663 and acyltransferase META1p3664 were firstly confirmed to catalyze the tailoring steps from 4,4'-diapolycopene-4,4'-dioic acid to carotenoid 1. In conclusion, the structure and biosynthetic genes of carotenoid 1 produced by M. extorquens AM1 were firstly characterized in this work, which shed lights on engineering M. extorquens AM1 for producing carotenoid 1 in high yield.

Comparison of Mitochondrial Genome Sequences between Two Palaemon Species of the Family Palaemonidae (Decapoda: Caridea): Gene Rearrangement and Phylogenetic Implications.

To further understand the origin and evolution of Palaemonidae (Decapoda: Caridea), we determined the mitochondrial genome sequence of Palaemon macrodactylus and Palaemon tenuidactylus. The entire mitochondrial genome sequences of these two Palaemon species encompassed 37 typical genes, including 13 protein-coding genes (PCGs), 2 ribosomal RNA genes (rRNAs), and 22 transfer RNA genes (tRNAs), and a control region (CR). The lengths of their mitochondrial genomes were 15,744 bp (P. macrodactylus) and 15,735 bp (P. tenuidactylus), respectively. We analyzed their genomic features and structural functions. In comparison with the ancestral Decapoda, these two newly sequenced Palaemon species exhibited a translocation event, where the gene order was trnK-trnD instead of trnD-trnK. Based on phylogenetic analysis constructed from 13 PCGs, the 12 families from Caridea can be divided into four major clades. Furthermore, it was revealed that Alpheidae and Palaemonidae formed sister groups, supporting the monophyly of various families within Caridea. These findings highlight the significant gene rearrangements within Palaemonidae and provide valuable evidence for the phylogenetic relationships within Caridea.

Sodium glucose cotransporter-2-inhibitor dapagliflozin improves nonalcoholic fatty liver disease by ameliorating dipeptidyl-peptidase-4 protein expression in diabetic mice.

INTRODUCTION: Nonalcoholic fatty liver disease (NAFLD) is the leading cause of chronic liver disease worldwide. It can progress from simple steatosis to nonalcoholic steatohepatitis and may even develop into liver fibrosis, hepatocirrhosis, or hepatocellular carcinoma, but there is no effective treatment. MATERIAL AND METHODS: Wild-type (wt) and diabetic (db/db) mouse NAFLD-induced models were used to investigate the hepatoprotective effects and potential mechanisms of dapagliflozin (a new oral hypoglycaemic drug) on type 2 diabetes mellitus (T2DM) complicated with NAFLD, and to establish wt and db/db mouse NAFLD-induced and dapagliflozin treatment models. RESULTS: Dapagliflozin reduces blood glucose, glycosylated haemoglobin, blood lipids, and serum transaminase levels in db/db mice and improves T2DM-related liver injury accompanied by NAFLD; the mechanism may be related to the decrease in dipeptidyl-peptidase-4 (DPP4) protein expression and improvement in liver enzymes. Further mechanism-related studies by our team revealed that dapagliflozin can also downregulate the expression of DPP4 proteins in the liver and reduce serum soluble DPP4 enzyme levels, thereby improving the hepatic steatosis and insulin resistance of NAFLD. CONCLUSION: Dapagliflozin may be an effective drug for the treatment of T2DM-induced NAFLD and NAFLD, providing a reliable laboratory basis and new treatment methods for the clinical treatment of NAFLD.

Label-free monitoring of spatiotemporal changes in the stem cell cytoskeletons in time-lapse phase-contrast microscopy.

Investigation of the dynamic structural changes in the actin cytoskeleton during cell migration provides crucial information about the physiological conditions of a stem cell during in-vitro culture. Here we proposed a quantitative analytical model associated with texture extraction with cell tracking techniques for in situ monitoring of the cytoskeletal density change of stem cells in phase-contrast microscopy without fluorescence staining. The reliability of the model in quantifying the texture density with different orientation was first validated using a series of simulated textural images. The capability of the method to reflect the spatiotemporal regulation of the cytoskeletal structure of a living stem cell was further proved by applying it to a set of 72 h phase-contrast microscopic video of the growth dynamics of mesenchymal stem cells in vitro culture.

Local neural-network-weighted models for occurrence and number of down wood in natural forest ecosystem.

The natural forest ecosystem has been affected by wind storms for years, which have caused several down wood (DW) and dramatically modified the fabric and size. Therefore, it is very important to explain the forest system by quantifying the spatial relationship between DW and environmental parameters. However, the spatial non-stationary characteristics caused by the terrain and stand environmental changes with distinct gradients may lead to an incomplete description of DW, the local neural-network-weighted models of geographically neural-network-weighted (GNNWR) models are introduced here. To verify the validity of models, our DW and environmental factors were applied to investigate of occurrence of DW and number of DW to establish the generalized linear (logistic and Poisson) models, geographically weighted regression (GWLR and GWPR) models and GNNWR (GNNWLR and GNNWPR) models. The results show that the GNNWR models show great advantages in the model-fitting performance, prediction performance, and the spatial Moran's I of model residuals. In addition, GNNWR models can combine the geographic information system technology for accurately expressing the spatial distribution of DW relevant information to provide the key technology that can be used as the basis for human decision-making and management planning.

Study on the Estimation of Forest Volume Based on Multi-Source Data.

We performed a comparative analysis of the prediction accuracy of machine learning methods and ordinary Kriging (OK) hybrid methods for forest volume models based on multi-source remote sensing data combined with ground survey data. Taking Larix olgensis, Pinus koraiensis, and Pinus sylvestris plantations in Mengjiagang forest farms as the research object, based on the Chinese Academy of Forestry LiDAR, charge-coupled device, and hyperspectral (CAF-LiTCHy) integrated system, we extracted the visible vegetation index, texture features, terrain factors, and point cloud feature variables, respectively. Random forest (RF), support vector regression (SVR), and an artificial neural network (ANN) were used to estimate forest volume. In the small-scale space, the estimation of sample plot volume is influenced by the surrounding environment as well as the neighboring observed data. Based on the residuals of these three machine learning models, OK interpolation was applied to construct new hybrid forest volume estimation models called random forest Kriging (RFK), support vector machines for regression Kriging (SVRK), and artificial neural network Kriging (ANNK). The six estimation models of forest volume were tested using the leave-one-out (Loo) cross-validation method. The prediction accuracies of these six models are better, with RLoo2 values above 0.6, and the prediction accuracy values of the hybrid models are all improved to different extents. Among the six models, the RFK hybrid model had the best prediction effect, with an RLoo2 reaching 0.915. Therefore, the machine learning method based on multi-source remote sensing factors is useful for forest volume estimation; in particular, the hybrid model constructed by combining machine learning and the OK method greatly improved the accuracy of forest volume estimation, which, thus, provides a fast and effective method for the remote sensing inversion estimation of forest volume and facilitates the management of forest resources.

Effect of Mechanical Heterogeneity on Strain and Stress Fields at Crack Tips of SCC in Dissimilar Metal Welded Joints.

The crack tip strain and stress condition are one of the main factors affecting stress corrosion cracking (SCC) behaviors in the dissimilar metal welded joint of the primary circuit in the pressurized water reactor. The mechanical property mismatch of base metal and weld metal can significantly affect the stress and strain condition around the crack tip. To understand the effect of different weld metals on strain and stress fields at SCC crack tips, the effects of strength mismatch, work hardening mismatch, and their synergy on the strain and stress field of SCC in the bi-material interface, including plastic zone, stress state, and corresponding J-integral, are investigated in small-scale yielding using the finite element method. The results show a significant effect of the strength mismatch and work hardening mismatch on the plastic zone and stress state in the weld metal and a negligible effect in the base metal. J-integral decreases with the single increase in either strength mismatch or work hardening mismatch. Either the increase in strength mismatch or work hardening mismatch will inhibit the other's effect on the J-integral, and a synthetic mismatch factor can express this synergistic effect.

Coculture, An Efficient Biotechnology for Mining the Biosynthesis Potential of Macrofungi via Interspecies Interactions.

Macrofungi, which are also known as mushrooms, can produce various bioactive constituents and have become promising resources as lead drugs and foods rich in nutritional value. However, the production of these bioactive constituents under standard laboratory conditions is inefficiency due to the silent expression of their relevant genes. Coculture, as an important activation strategy that simulates the natural living conditions of macrofungi, can activate silent genes or clusters through interspecific interactions. Coculturing not only can trigger the biosynthesis of diverse secondary metabolites and enzymes of macrofungi, but is also useful for uncovering the mechanisms of fungal interspecific interactions and novel gene functions. In this paper, coculturing among macrofungi or between macrofungi and other microorganisms, the triggering and upregulation of secondary metabolites and enzymes, the potential medicinal applications, and the fungal-fungal interaction mechanisms are reviewed. Finally, future challenges and perspectives in further advancing coculture systems are discussed.

Role of HPV status and PD-L1 expression in prognosis of laryngeal squamous cell carcinoma.

PURPOSE: Human papillomavirus (HPV) infection has been recognized as a cause of head and neck squamous cell carcinomas (HNSCC). Laryngeal squamous cell carcinoma (LSCC) is one of the most common pathologic types of HNSCC. Clinical trials show that there are differences in response to immunotherapy according to HPV status. It was reported that a high level of programmed cell death-ligand 1 (PD-L1) is correlated with better survival in HPV-positive head and neck cancer. In this study, we investigated the expression of PD-L1 in HPV-positive and HPV-negative LSCC to determine its prevalence and prognostic value. METHODS: 52 cases of LSCC were collected from Tangshan Head and Neck Disease Pathology Research Base. PCR-reverse dot blot hybridization and RNAscope in situ hybridization were used to detect HPV status. PD-L1 expression was evaluated by immunohistochemistry and all cases were followed up for survival. SPSS24.0 was used for data entry and statistical analysis. Kaplan-Meier method and Log-rank time series analysis were used for single factor analysis. Multivariate analysis was performed using Cox proportional hazard regression model, and HR and 95% CI were calculated. RESULTS: Of the 52 LSCC patients, 32.7% (17/52) were HPV-positive by RNAscope in situ hybridization, and 51.9% (27/52) of patients were positive for PD-L1 expression by immunohistochemistry. Regression analysis showed that with a median follow-up period of 69 months, smoking and late stage were associated with poor overall survival (OS), whereas HPV positivity and PD-L1 expression showed a better overall survival outcome. CONCLUSION: Smoking status, tumor stage, HPV status, and PD-L1 expression in tumor cells may represent useful prognostic biomarkers in patients with LSCC.

Histone deacetylase inhibitor valproic acid attenuates high glucose­induced endoplasmic reticulum stress and apoptosis in NRK­52E cells.

Previous studies have demonstrated that valproic acid (VPA), a histone deacetylase inhibitor, alleviates diabetic nephropathy (DN). However, the biological mechanisms underlying this protective effect remains unclear. This study aimed to investigate the effects of histone deacetylase inhibitor VPA on hyperglycemic induction of NRK­52E cell ERS and apoptosis. Endoplasmic reticulum stress (ERS)­related apoptosis is involved in DN, and improving ERS may delay the symptoms of DN. Histone deacetylase regulates gene transcription or expression of ERS­related proteins. The present study established an ERS model by treating the rat renal tubular epithelial cells NRK­52E with high glucose (HG) and investigated the effects of VPA on the apoptosis of the NRK­52E cells. HG stimulation significantly increased the protein levels of the ERS­related proteins including glucose regulated protein 78 (GRP78), activating transcription factor 4 (ATF4), C/EBP homologous protein (CHOP), caspase­12 and phosphorylated (p)­JNK. VPA treatment further upregulated GRP78 expression and attenuated the levels of ATF4, CHOP, caspase­12 and p­JNK. Notably, HG markedly promoted apoptosis of NRK­52E cells by regulating the protein levels of Bax, cleaved caspase­3 and Bcl­2, which was attenuated by simultaneous VPA treatment. Mechanistically, VPA increased the total acetylation levels of histone H4 in NRK­52E cells and increased the histone H4 acetylation of the GRP78 promoter region. In conclusion, VPA attenuated HG­induced ERS and apoptosis in NRK­52E cells, which may be due to the regulation of acetylation levels of ERS­related proteins. In addition, the present study suggested that HDACIs are promising drugs for treating patients with DN.

High risk HPV detection by RNAscope in situ hybridization combined with Cdc2 protein expression by immunohistochemistry for prognosis of oropharyngeal squamous cell carcinoma.

High risk human papillomavirus (HPV) infection is related to the development of head and neck squamous cell carcinoma (HNSCC). Oropharyngeal squamous cell carcinoma (OPSCC) is a common type of HNSCC, and its incidence has increased significantly in recent years. In this study, high risk HPV, the expression of P53, P21, and Cdc2 in OPSCC tissues was detected and the prognostic factors and clinical value of OPSCC were discussed. According to the WHO classification and diagnosis standard for head and neck tumors (2017 Edition), 49 OPSCC cases with complete clinical data were collected from Tangshan Head and Neck Disease Pathology Research Base from January 1, 2012 to December 31, 2018. The E6 and E7 mRNA of HPV 16 and HPV 18 were detected by RNAscope in situ hybridization. The expression of P53, P21, and Cdc2 protein was observed by SP immunohistochemical method and all cases were followed up for survival. Median survival time was analyzed by Kaplan-Meier method. The Log-rank test was used for single factor analysis and Cox regression model was used to analyze multiple prognostic factors. In 49 OPSCC cases the median age was 53 years; 14 were HPV-DNA positive (14/49, 28.6%) while 35 were negative (35/49, 71.4%). E6, E7 mRNA test showed that 20 cases (20/49, 40.8%) were positive for HPV-16. Among them 11 cases were positive for HPV-16 DNA. 2 cases were positive for HPV-18 mRNA (2/49, 4.08%). 27 cases were negative for mRNA16 and 18 (27/49, 55.1%). The prevalence of HPV was 68.8% (11/16) in the non-smoking group, which was higher than that of the smoking group (10/33, 33.3%), (χ2=5.463, P=0.019). There was no significant correlation between HPV detection and gender, age, drinking, tumor differentiation degree, and clinical stage (P > 0.05). The expression rates of P53, P21, and Cdc2 in OPSCC tissues were 63.3% (31/49), 65.3% (32/49), and 67.3% (33/49), respectively. There was no significant correlation between expression of all the three proteins and gender, age, HPV, smoking, drinking, tumor differentiation, and clinical stage (P > 0.05). Cox multifactor regression analysis showed that HPV (HR=0.275, 95% CI: 0.146-0.517), tumor differentiation (HR=1.751, 95% CI: 1.231-2.492), stage (HR=3.268, 95% CI: 1.758-6.074) and expression of Cdc2 protein (HR=1.804, 95% CI: 0.990-3.286) were related to the survival time of patients (P < 0.05). Our findings support that most of the HPV-positive OPSSC patients were non-smokers. The patients with negative HPV, low differentiation, late stage, and Cdc2 positive expression have poor prognosis and need to be followed up.

Quantitative Bioimage Analysis of Passaging Effect on the Migratory Behavior of Human Mesenchymal Stem Cells During Spheroid Formation.

The quality of stem cells obtained through serial subcultivation is the pivotal factor determining the therapeutic effectiveness of regenerative medicine. However, an effective quality monitoring system for cell culture is yet to be established. Detailed parameter studies of the migratory behavior of stem cells at different passages may provide insight into the deterioration of stemness. Thus, this study aimed to evaluate the feasibility of quantitative bioimage analysis for monitoring stem cell quality during in vitro culture and to explore the passaging effects on stem cell migration. An image-based analytical tool using cell tracking, cytometric analyses, and gating with time-lapse microscopy was developed to characterize the migratory behavior of human mesenchymal stem cells (hMSCs) isolated from human adipose tissue (hADAS) and placenta (hPDMC) cultured on chitosan membranes. Quantitative analysis was performed for the single cells and assembled spheroids selected from 15 videos of Passages 3, 5, and 11 for hADAS and those from 12 videos of Passages 7, 11, and 16 for hPDMC. These passages were selected to represent the young, matured, and degenerated stem cells, respectively. Migratory behavior varied with cell passages. The mobility of single hMSCs decreased at degenerated passages. In addition, enhancement of mobility, due to transformation from single cells to spheroids, occurred at each passage. The young hMSCs seemed more likely to move as single cells rather than as aggregates. Once matured, they tended to aggregate with strong 3D spheroid formability and increased mobility. However, the spheroid formability and mobility decreased at late passage. The increase in aggregation rate with passaging may be a compensatory mechanism to enhance the declining mobility of hMSCs through cell coordination. Our findings regarding the passaging effects on stem-cell migratory behavior agree with biochemical reports, suggesting that the developed imaging method is capable of monitoring the cell-culture quality effectively. © 2020 International Society for Advancement of Cytometry.

A dual-label time-resolved fluorescence immunoassay for the simultaneous determination of carcinoembryonic antigen and squamous cell carcinoma antigen.

Among women worldwide, cervical cancer is the second-most common cancer, and cervical smears and DNA detection have low sensitivity or are too expensive. The concentrations of carcinoembryonic antigen (CEA) and squamous cell carcinoma (SCC) in the serum were detected using a sandwich immunoassay. The CEA and SCC in the serum were captured by anti-CEA and anti-SCC antibodies. After combining other anti-CEA- and anti-SCC-labeled antibodies with europium (III) (Eu3+ ) and samarium (III) (Sm3+ ) chelates, CEA and SCC were detected with time-resolved fluorometry (TRF). The linear correlation coefficients (R2 ) of the CEA and SCC standard curves were 0.9997 and 0.9997, respectively. The minimum detection level for CEA was 1.15 ng/mL (the linear dynamic range was 3.24-543.67 ng/mL), and the average recovery was 100.83%. The sensitivity for SCC detection was 0.54 ng/mL (the linear dynamic range was 2.47-96.58 ng/mL), and the average recovery was 101.02%. High R2 between the results of commercial assays and this method were obtained (R2  = 0.9983 for CEA, R2   = 0.9878 for SCC). These findings indicated that the dual-label TRFIA invented in this study has high sensitivity, accuracy, and specificity in clinical analysis, which indicates that this method could be used for the early diagnosis and follow-up surveillance of cervical cancer.

Prognostic value of human papillomavirus infection and p53, p16, epidermal growth factor receptor and p34cdc2 expression in patients with salivary adenoid cystic carcinoma.

The aim of this study was to explore the correlation between HPV infection, p53, p16, epidermal growth factor receptor (EGFR), p34cdc2 protein expression and prognosis in patients with adenoid cystic carcinoma of salivary gland. Totally 78 cases of adenoid cystic carcinoma of salivary gland specimens were selected from January 1, 2004 to December 31, 2013 in Tangshan Union Hospital. PCR-reverse dot blot hybridization was used to detect infection of human papilloma virus (HPV), and SP immunohistochemical method was adopted to detect the expression of p53, p16, EGFR and p34cdc2 protein in the carcinoma tissues. Clinical data were collected and the patients were followed up. Results showed that the infection rate of HPV in adenoid cystic carcinoma tissues was 0% (0/78). The expression rate of p53, p16, EGFR and p34cdc2 protein in carcinoma tissues were 75.6% (59/78), 57.7% (45/78), 60.1% (47/78) and 64.1% (50/78), respectively. Expression of p53, p16, EGFR and p34cdc2 proteins was not significantly correlated with patients' age, gender, disease location, TNM classification and histological type (P > 0.05). Kaplan-Meier analysis showed that EGFR-positive patients had a lower median overall survival than EGFR-negative ones (58 months vs. 75 months, respectively. P = 0.001). The result of median progression-free survival was virtually the same for both EGFR-positive and EGFR-negative patients (43 months vs. 49 months, respectively. P = 0.002). p34cdc2-positive patients had a lower median overall survival than p34cdc2-negative ones (61 months vs. 71 months, respectively. P = 0.027). Median progression-free survival was also almost the same for both p34cdc2-positive and p34cdc2-negative patients (44 months vs. 51 months, respectively. P = 0.011). Cox regression analysis showed that expression of EGFR and p34cdc2 was independent risk factors for the prognosis of patients with adenoid cystic carcinoma of salivary gland (relative risk = 13.199, 11.466, P < 0.001). In conclusion, HPV infection is not detected in salivary adenoid cystic carcinoma tissues. p53, p16, EGFR and p34cdc2 protein are positively expressed in most salivary adenoid cystic carcinoma tissues. p16 is unsuitable as a surrogate for HPV infection status in patients with adenoid cystic carcinoma of salivary gland. Expression of EGFR and p34cdc2 is independent risk factors in the prognosis of patients with salivary gland adenoid cystic carcinoma. Patients with EGFR or p34cdc2 positive expression should be followed up closely.

Expression of p27 and PTEN and clinical characteristics in early laryngeal squamous cell carcinoma and their correlation with recurrence.

Laryngeal squamous cell carcinoma is a common malignant tumor of otolaryngeal region. At present, effective treatment of laryngeal squamous cell carcinoma still depends on surgery and radiotherapy. In recent years, application of CO2 laser resection in the treatment of stage T1 glottic carcinoma can remove the tumor completely and reduce the injury of laryngeal tissues. But recurrence still happened in some postoperative patients. Here, we selected 131 patients to compare the therapeutic effects of CO2 laser resection and traditional split laryngeal surgery on the early laryngeal cancer, examined the expression of p27 and PTEN by immunohistochemistry in early laryngeal squamous cell carcinoma tissues in correlation to clinical outcome. After two years follow-up 14/85 (16.5%) of CO2 laser treatment group presented with local recurrence (recurrent group), while that of split laryngeal surgery group was 6/46 (13.0%). There was no statistical significance in recurrence rate between the two groups (P>0.05). 10 of all the 111 (9.0%) non-recurrent patients did not follow the doctor's advice to quit smoking after the operation, while 12 in the 20 (60.0%) recurrent patients did not; the difference between the two groups was statistically significant (P<0.01). The positive rates of p27 were 80.2% (105/131) and 43.5% (57/131), and that of PTEN were 83.2% (109/131) and 48.9% (64/131) in the cancer adjacent tissues (negative surgical margin tissues) and in laryngeal carcinoma tissues, respectively (P<0.001). The expression rates of p27 and PTEN in laryngeal carcinoma tissues of the recurrent group were 20.0% (4/20), 10.0% (2/20) and that in non recurrent group were 47.7% (53/111) and 55.9% (62/111), respectively, with a significant difference (P<0.001). In addition, the expression of p27 and PTEN in tumor resected marginal tissues of the recurrence group was 50.0% (10/20), 40.0% (8/20) and that in non recurrence group was 85.6% (95/111) and 91.0% (101/111), respectively; the difference was also statistically significant between both groups (P<0.001). In conclusion, there is no statistically significant difference in tumor recurrence rate between CO2 laser surgery and traditional split laryngeal surgery. Postoperative recurrence is closely related to resume smoking. The recurrence rate of p27 and/or PTEN-negative patients was higher than that of the positive ones,that should be followed up closely after treatment.

Over-expression of p53, p21 and Cdc2 in histologically negative surgical margins is correlated with local recurrence of laryngeal squamous cell carcinoma.

The aim of this study was to explore the relationship between the expression of p53, p21 and Cdc2 in the early laryngeal cancer with negative pathological margins and its local recurrence. During 2004-2010, a total of 85 patients with early laryngeal cancer were selected in Tangshan Union Hospital, Hebei, China, and immunohistochemical method was used to detect the expression of p53, p21 and Cdc2 in the negative pathological margin tissues. All patients were followed up for two years to collect pathological data for evaluating the survival and tumor recurrence. Two years after surgery 14 of 85 patients with laryngeal cancer presented with recurrence (recurrent group), while 71 patients without recurrence (none recurrent group). The positive rate of p53, p21 and Cdc2 protein in laryngeal cancer tissues was 60.0% (51/85), 38.8% (33/85) and 70.6% (60/85), respectively, while that of the three proteins in the cancer adjacent tissues was 36.5% (31/85), 21.2% (18/85) and 29.4% (25/85), respectively. The differentiation and TNM stage of tumor had no correlation with the three proteins. The positive rate of p53 in the surgical margin of the recurrent group and non recurrent group was 71.4% (10/14) and 29.6% (21/71) (P = 0.003), that of p21 was 50.0% (7/14) and 15.5% (11/71), (P = 0.011) and Cdc2 was 57.1% (8/14) and 23.9% (17/71) (P = 0.030), respectively. In conclusion, p53, p21 and Cdc2 may be involved in the occurrence, development and recurrence of laryngeal squamous cell carcinoma. Overexpression of p53, p21 and Cdc2 in the surgical margin of early laryngeal cancer is closely related to local recurrence of tumor.

Down-regulation of microRNA-181b is a potential prognostic marker of non-small cell lung cancer.

The aim of this study was to investigate the clinical significance of microRNA-181b (miR-181b) expression in non-small cell lung cancer (NSCLC). MiR-181b expression in 126 pairs of surgically removed NSCLC tissues and their corresponding normal lung tissues was measured by real-time quantitative RT-PCR assay. Additionally, the correlation of miR-181b expression with clinicopathological factors or prognosis of patients was analyzed. At first, miR-181b expression was significantly down-regulated in NSCLC tissues as compared with their normal counterparts (P<0.001). Then, the low miR-181b expression was found to be closely correlated with larger tumor size (P=0.02), higher p-TNM stage (P=0.008) and positive lymph node metastasis (P=0.03) of NSCLC patients. After that, survival analysis found that the overall survival (P=0.001) and disease-free survival (P=0.008) of NSCLC patients with low miR-181b expression were both significantly poorer compared to those patients with high miR-181b expression. Finally, both univariate and multivariate analyses demonstrated that low miR-181b expression may be a poor prognostic marker of NSCLC patients. This is the first study to indicate that down-regulation of miR-181b may be correlated with aggressive disease progression and poor prognosis of NSCLC patients, suggesting that miR-181b might be involved in lung carcinogenesis and become a potential prognostic marker for NSCLC.

[Study of SOD and PON-1 expression in type 2 diabetes mellitus nephropathy and its clinical significance].

AIM: To investigate the clinical value of superoxide dismutase (SOD) and paraoxonase-1 (PON-1) in type 2 diabetes mellitus (T2DM) nephropathy. METHODS: 80 cases of T2DM patients were divided into T2DM nephropathy group (ND group, 40 cases), and non-T2DM nephropathy group (non-ND group, 40 cases), the SOD, PON-1 levels and biochemical levels of the two group were measured, the HOMA-IR was recorded. RESULTS: The SOD and PON-1 levels of the ND group were lower than the non-ND group, compared the difference was significant (P < 0.05). The FINS, FINS levels and HOMA-IR of the ND group were higher than the non-ND group, compared the difference was significant (P < 0.05). The FBG, PBG2h and HbAlc levels were no significantly different (P > 0.05). Serum SOD, PON-1 levels and HOMA-IR were significantly negatively correlated (r = -0.287, -0.509; P = 0.006, 0.000), serum SOD level and PON-1 level was no significantly correlated (P > 0.05). CONCLUSION: The serum SOD and PON-1 involve in the development of T2DM nephropathy. Combined detection of serum SOD and PON-1 have important clinical significance for early diagnosis, prevention, treatment and prognosis of T2DM nephropathy.