Biomarker Screening by LCMS and Liquid Chip Technology in Acute Aortic Dissection.

BACKGROUND: Aortic dissection (AD) is a serious disease. Previous study, the use of peripheral blood biomarkers to diagnose AD showed strong clinical feasibility, but the possible molecular mechanism is unclear. METHODS: Sera from 79 healthy subjects, 73 patients with well-established AD, and 74 patients with well-established acute myocardial infarction (AMI) were investigated by Liquid Chromatograph-Mass Spectrometer to detect metabolites (AFMK, Glycerophosphocholine, Inosine, SPH). The cell factor expression in the 3 group were detected by Liquid Chip Technology. RESULTS: The serum content trends of 4 metabolic indexes in patients with AMI and AD group were used as the diagnostic models, and the effective diagnosis rate was 97.8%. The diagnosis rate is 89.8% in distinguishing patients with AMI from patients with AD. The expression in serum of the 3 groups showed that there were significant differences in the expression of 23 cytokines. By correlation analysis, it was found that miP-1, IL-7, MIP-1ß, EGF and other cytokines were significantly correlated with the 4 metabolic molecules. CONCLUSIONS: AFMK, Glycerophosphocholine, Inosine, Sphingfungin B (SPH) metabolites are potential biomarkers for AD, and the influence of related metabolic process may be related to the expression of miP-1, IL-7, MIP-1ß, EGF, and other cytokines.

Retracted: Long Noncoding RNA TP73-AS1 Targets MicroRNA-329-3p to Regulate Expression of the SMAD2 Gene in Human Cervical Cancer Tissue and Cell Lines.

The Editors of Medical Science Monitor wish to inform you that the above manuscript has been retracted from publication due to concerns with the credibility and originality of the study, the manuscript content, and the Figure images. Reference: Mei Mei Guan, Qun Xian Rao, Miao Ling Huang, Li Juan Wang, Shao Dan Lin, Qing Chen, Chang Hao Liu. Long Noncoding RNA TP73-AS1 Targets MicroRNA-329-3p to Regulate Expression of the SMAD2 Gene in Human Cervical Cancer Tissue and Cell Lines. Med Sci Monit, 2019; 25: 8131-8141. DOI: 10.12659/MSM.916292.

Are Early Embryo Cleavage Kinetics Affected by Energy Substrates in Different Culture Media?

OBJECTIVE: This study aimed to investigate the influence of different culture media on early embryonic cleavage kinetics using time-lapse analysis and to determine the possible relationships between energy substrates in culture media and the cleavage kinetics. METHODS: A total of 10 021 embryos from 1310 couples were cultured in time-lapse incubators. Embryos cultured in Vitrolife media were allocated to group I, and those in COOK media to group II. Embryo cleavage time points up to the 8-cell stage (t2-t8) were observed after pronuclei fading. RESULTS: The baseline demographic features, in vitro fertilization indications, ovarian stimulation protocol, oocyte-cumulus complexes, fertilization rate, together with pregnancy and perinatal outcomes were similar (P>0.05) between groups I and II. According to the time-lapse analysis, all embryos in group I showed significantly faster cleavage speed than those in group II (P<0.05). Furthermore, there was better synchrony in division (s3) and a longer length of the third cell cycle duration (cc3) in group II. Interestingly, implanted embryos in group II showed faster cleavage speed than those in group I, especially at t4 and t7. The glucose contents and multiple major amino acids were similar between the two groups. Lactic and pyruvic acid contents were generally higher in group I than those in group II. CONCLUSION: Because different commercial culture media may influence cleavage kinetics of embryos, it is essential for embryologists to take culture media into consideration in selecting a potential embryo when using a time-lapse system before implantation.

Transformer Winding Deformation Detection Based on BOTDR and ROTDR.

In order to realize distributed measurement of transformer winding temperature and deformation, a transformer winding modification scheme with a built-in distributed optical fiber was designed. By laying a single-mode fiber and a multi-mode fiber on the transformer winding, the Brillouin optical time domain reflection technique (BOTDR) and the Raman optical time domain reflection technique (ROTDR) are used to measure the strain and temperature of the winding to complete the more accurate winding deformation detection. The accuracy of strain and temperature sensing of this scheme was verified by simulation. Then, according to the scheme, a winding model was actually wound, and the deformation and temperature rise tests were carried out. The test results show that this scheme can not only realize the deformation detection and positioning of the winding, but can also realize the measurement of the winding temperature; the temperature measurement accuracy reached ±0.5 °C, the strain measurement accuracy was 200 µÎµ, and spatial resolution was up to 5 m. In this experiment, the deformation location with the precision of 2 turns was realized on the experimental winding.

Long Noncoding RNA TP73-AS1 Targets MicroRNA-329-3p to Regulate Expression of the SMAD2 Gene in Human Cervical Cancer Tissue and Cell Lines.

BACKGROUND Worldwide, mortality from cervical cancer in women remains high. This study aimed to investigate the expression of long noncoding RNA (lncRNA) TP73-AS1, microRNA-329-3p (miRNA-329-3p), and the SMAD2 gene and their regulatory relationships in human cervical cancer tissue and cervical cancer cell lines. MATERIAL AND METHODS Cervical cancer tissue samples (n=30) and normal control cervical tissues were studied. Cell proliferation and migration were investigated in HeLa and SiHa human cervical cancer cells using the MTT assay, crystal violet staining, wound healing assay, and the transwell assay. Expression of lncRNA TP73-AS1 and the SMAD2 gene were detected using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Enrichment of miR-329-3p was measured using the RNA immunoprecipitation assay (RIPA). Targeting relationships between TP73-AS1, miR-329-3p, and SMAD2 were identified using the dual-luciferase reporter assay. A subcutaneous xenograft model was established, tumor size was measured, and SMAD2 expression was detected using immunohistochemistry. RESULTS LncRNA TP73-AS1 was overexpressed in cervical cancer tissues and cells and was associated with reduced expression of miR-329-3p. Down-regulation of lncRNA TP73-AS1 inhibited cell proliferation, migration and invasion and increased miR-329-3p expression. Expression of SMAD2 down-regulated miR-329-3p and was associated with increased expression of TP73-AS1. LncRNA TP73-AS1 knockdown resulted in miR-329-3p silencing. In tumor xenografts, expression of TP73-AS1 reduced the tumor volume and down-regulated the expression levels of the SMAD2 gene. CONCLUSIONS LncRNA TP73-AS1 promoted proliferation of cervical cancer cell lines by targeting miR-329-3p to regulate the expression of the SMAD2 gene. A regulatory network was formed between lncRNA TP73-AS1, miR-329-3p, and SMAD2.

The added value of fasting blood glucose to serum squamous cell carcinoma antigen for predicting oncological outcomes in cervical cancer patients receiving neoadjuvant chemotherapy followed by radical hysterectomy.

OBJECTIVE: To determine the combination of fasting blood glucose (FBG) with squamous cell carcinoma antigen (SCCA) assessments in the prediction of tumor responses to chemotherapy and pretreatment prognostication among patients receiving neoadjuvant chemotherapy (NACT) for locally advanced cervical cancer (LACC). METHODS: Data of 347 LACC patients were retrospectively reviewed. Receiver operating characteristic (ROC) curves were constructed, and areas under the curves (AUCs) were compared to evaluate the ability to predict complete response (CR) following NACT. Patients were stratified into groups with low and high levels of SCCA and FBG and combined into low- or high-SCCA and low- or high-FBG groups. Cox regression analysis was performed to identify determinants of recurrence-free survival (RFS) and overall survival (OS). RESULTS: The AUCs were 0.70, 0.68, and 0.66 for SCCA, FBG, and a combination of SCCA and FBG for predicting CR following NACT, respectively; however, the differences among AUCs were not significant (P = .496). Pretreatment SCCA and FBG levels were identified as independent predictors of RFS and OS. The high-SCCA/high-FBG group showed significantly worse prognosis than the low-SCCA/low-FBG group. After adjusting for other variables, high-SCCA/high-FBG remained independently associated with an increased risk of tumor recurrence and death. CONCLUSION: SCCA, FBG, and a combination of SCCA and FBG could acceptably predict CR following NACT. Pretreatment SCCA and FBG levels were independent prognostic factors. The combination of SCCA and FBG levels refined the prognostic stratification of LACC patients, which allowed the group of patients with the highest risk of recurrence and death to be identified.

Associations of human leukocyte antigen and interleukin-18 gene polymorphisms with viral load in patients with hepatitis B infection.

This study aimed to assess the associations of human leukocyte antigen (HLA)-DR and interleukin (IL)-18 gene polymorphisms with hepatitis B virus (HBV).Clinical data were retrospectively reviewed between December 2006 and December 2015 at Xiangyang Central Hospital. HBV patients were assigned to the high and low viral load groups, respectively, according to HBV copies. HLA-DRB1*03 polymorphisms and IL-18 polymorphisms were detected by sequence-specific primer-polymerase chain reaction (PCR-SSP) and PCR-ligase detection reaction (PCR-LDR), respectively. T cell subgroups were identified by flow cytometry, and IL-18, IL-12, interferon-γ (IFN-γ), IL-4, and IL-10 expression levels were assessed by ELISA. A total of 630 subjects were included in the analysis.Compared with healthy controls, the chronic HBV group showed significantly lower IL-18 (P < .001), IL-12 (P < .001), and IFN-γ (P < .001) expression levels, and markedly increased IL-4 (P < .001) and IL-10 (P < .001) amounts. Th2 cytokine expression was high in HLA-DRB1*03 positive (+) HBV patients, with low Th1 cytokine levels. The ratios of CD4+/CD8+ and Th1/Th2 cells decreased with increasing HBV DNA levels. The chronic HBV group showed a relatively high frequency of -137G in the IL-18 gene, while IL-18 expression was low in homozygous GG genotype individuals.Polymorphisms in the HLA-DRB1*03 and IL-18 genes are associated with viral load in HBV. HLA-DRB1 and IL-18 gene polymorphisms are involved in the regulation of the Th1/Th2 balance and expression of relevant cytokines that influence immune responses in HBV.

Leinal polypeptide promotes NK cells to suppress PCa survival in vitro.

As an important component in the innate immune system, natural killer (NK) cells have been demonstrated to be clinically associated with prostate cancer (PCa) progression and castration resistance. Therefore, the development of novel agents that may enhance the cytotoxicity of NK cells possesses promising therapeutic applications. In the present study, leinal polypeptide (LP) solution was supplemented into a co-culture system of NK and PCa cells, as it was previously demonstrated that LP are able to activate NK cells, which kill PCa cells based on an MTT cell viability assay. Mechanistic dissection demonstrated that LP enhanced androgen receptor degradation, which resulted in an upregulation of MHC class I polypeptide-related sequence A (MICA) and MICB. In turn, the induced expression of MICA and MICB was able to further trigger NK cell activation, forming a positive loop between NK cells and PCa cells in the presence of LP solution.

Pretreatment glycemic control status is an independent prognostic factor for cervical cancer patients receiving neoadjuvant chemotherapy for locally advanced disease.

BACKGROUND: To investigate whether poor glycemic control status has a negative impact on survival outcomes and tumor response to chemotherapy in patients receiving neoadjuvant chemotherapy (NACT) for locally advanced cervical cancer (LACC). METHODS: A retrospective cohort study was conducted to examine LACC patients undergoing NACT and radical hysterectomy between 2002 and 2011. Patients were divided into three groups: patients without diabetes mellitus (DM), diabetic patients with good glycemic control, and diabetic patients with poor glycemic control. Hemoglobin A1c (HbA1c) levels were used to indicate glycemic control status. Recurrence-free survival (RFS), cancer-specific survival (CSS) and overall survival (OS) were analyzed using log-rank tests and Cox proportional hazards models. RESULTS: In total, 388 patients were included and had a median follow-up time of 39 months (range: 4-67 months). Diabetes mellitus (DM) was diagnosed in 89 (22.9%) patients, only 35 (39.3%) of whom had good glycemic control prior to NACT (HbA1c < 7.0%). In survival analysis, compared with patients with good glycemic control and patients without DM, patients with poor glycemic control (HbA1c ≥ 7.0%) exhibited decreased recurrence-free survival (RFS), cancer-specific survival (CSS) and overall survival (OS). In multivariate analysis, HbA1c ≥ 7.0% was identified as an independent predictor for decreased RFS (hazard ratio [HR] = 3.33, P < 0.0001), CSS (HR = 3.60, P < 0.0001) and OS (HR = 4.35, P < 0.0001). In the subgroup of diabetic patients, HbA1c ≥ 7.0% prior to NACT had an independent negative effect on RFS (HR = 2.18, P = 0.044) and OS (HR = 2.29, P = 0.012). When examined as a continuous variable, the HbA1c level was independently associated with decreased RFS (HR = 1.39, P = 0.002), CSS (HR = 1.28, P = 0.021) and OS (HR = 1.27, P = 0.004). Both good (odds ratio [OR] = 0.06, P < 0.0001) and poor glycemic control (OR = 0.04, P < 0.0001) were independently associated with a decreased likelihood of complete response following NACT. CONCLUSIONS: Poor glycemic control is an independent predictor of survival and tumor response to chemotherapy for patients receiving NACT for LACC.

miR-34C Disrupts the Stemness of Purified CD133+ Prostatic Cancer Stem Cells.

OBJECTIVE: To find the potential micro-RNA (miRNA) that could determine the fate of prostate cancer stem cells. MATERIALS AND METHODS: We compared miRNA expression between our purified CD133+ prostatic cancer stem cells (PCSCs) and CD133- cells. Sphere formation assay and matrigel-based cell invasion assay were applied to determine the stemness of CD133+ PCSCs after our manipulation of miRNA using miRNA mimic or miRNA inhibitor. RESULTS: In this study, we identified that miR-34C was under-expressed in the purified CD133+ PCSCs and enforced introduction of miR-34C attenuated the stemness of CD133+ PCSCs. Clinically, we also observed a negative correlation between miR-34C and CD133. CONCLUSION: Our data strongly suggest that miR-34C may play essential role in conferring castration resistance by equilibrating PSCS population.

Recruited mast cells in the tumor microenvironment enhance bladder cancer metastasis via modulation of ERß/CCL2/CCR2 EMT/MMP9 signals.

Early clinical studies suggested that infiltrating mast cells could be associated with a poor outcome in bladder cancer (BCa) patients. The mechanisms of how mast cells influence the BCa progression, however, are unclear. Using the human clinical BCa sample survey and in vitro co-culture systems, we found BCa cells could recruit more mast cells than the surrounding non-malignant urothelial cells. The consequences of this better recruitment of mast cells toward BCa cells could then enhance BCa cell invasion. Mechanism dissection revealed that the enhanced BCa cell invasion could function via up-regulation of the estrogen receptor beta (ERß) in both mast cells and BCa cells, which resulted in the increased CCL2/CCR2/EMT/MMP9 signals. Using the pre-clinical mouse BCa model, we further validated the mast cell-promoted BCa invasion. Interruption of the newly identified ERß/CCL2/CCR2/EMT/MMP9 pathway via either ERß-siRNA, ERß antagonist PHTPP, or CCR2 antagonist can effectively reverse the mast cell-enhanced BCa cells invasion. Together, our finding could lead to the development of an alternative new therapeutic approach to better treat BCa metastasis.

Androgen receptor (AR) suppresses miRNA-145 to promote renal cell carcinoma (RCC) progression independent of VHL status.

Mutational inactivation of the VHL tumor suppressor plays key roles in the development of renal cell carcinoma (RCC), and mutated VHL-mediated VEGF induction has become the main target for the current RCC therapy. Here we identified a signal pathway of VEGF induction by androgen receptor (AR)/miRNA-145 as a new target to suppress RCC progression. Mechanism dissection revealed that AR might function through binding to the androgen receptor element (ARE) located on the promoter region of miRNA-145 to suppress p53's ability to induce expression of miRNA-145 that normally suppresses expression of HIF2α/VEGF/MMP9/CCND1. Suppressing AR with AR-shRNA or introducing exogenous miRNA-145 mimic can attenuate RCC progression independent of VHL status. MiR-145 mimic in preclinical RCC orthotopic xenograft mouse model revealed its efficacy in suppression of RCC progression. These results together identified signals by AR-suppressed miRNA-145 as a key player in the RCC progression via regulating HIF2α/VEGF/MMP9/CCND1 expression levels. Blockade of the newly identified signal by AR inhibition or miRNA-145 mimics has promising therapeutic benefit to suppress RCC progression.

[The prediction value of noninvasive bioreactance-based passive leg raising test in assessing fluid responsiveness in elderly patients with sepsis].

OBJECTIVE: To investigate the value of bioreactance-based passive leg raising (PLR) test predicting fluid responsiveness of elderly patients with sepsis. METHODS: This prospective and self-controlled clinical study included 31 elderly patients with sepsis in the Department of Intensive Care Medicine of Zhejiang Hospital. Hemodynamic parameters including cardiac output (CO), stroke volume variation (SVV) were continuously recorded by bioreactance-based device (noninvasive cardiac output monitoring, NICOM) before and after PLR and volume expansion (VE) test. Patients were defined as responders if CO increased ≥ 10% after VE. RESULTS: A total of 100 PLR and VE tests in these 31 patients were evaluated.In 28 responders, CO[(5.11 ± 2.10) L/min vs (5.91 ± 2.45) L/min, P < 0.05; (5.06 ± 2.06) L/min vs (5.77 ± 2.47) L/min, P < 0.05] and SV [(59.61 ± 18.22) ml vs (69.29 ± 21.32) ml, P < 0.05; (60.10 ± 15.95) ml vs (70.06 ± 17.96) ml, P < 0.05] were obviously increased both after PLR and VE. The ΔCO after PLR (ΔCOPLR) and ΔCOVE was highly correlated (r = 0.819, P = 0.001) while the SVV before VE and Δ COVE was uncorrelated (r = -0.218, P = 0.059). The areas under the ROC curve of ΔCOPLR, SVV predicting fluid responsiveness were 0.859 and 0.459 respectively. The ΔCOPLR ≥ 10% was found to predict fluid responsiveness with a sensitivity and specificity of 85% and 83% respectively. CONCLUSION: Compared with SVV, PLR test is a simple, effective method for accurately predicting fluid responsiveness of elderly patients with sepsis.

[Effects of postoperative restrictive fluid management on recovery of gastrointestinal function after elective colonic resection].

OBJECTIVE: To investigate the effect of postoperative restrictive fluid management by ensuring adequate tissue perfusion on the recovery of gastrointestinal function after elective colonic resection. METHODS: Thirty patients suffered with elective colonic resection, after 6 hours of anesthesia recovery, were randomly divided into restrictive fluid management group (restrictive group, n=15) and traditional fluid management group (control group, n=15). From the surgery day to the 4th postoperative day, patients in restrictive group and control group received the total fluids of 25-35 ml×kg(-1)×d(-1) or 40-50 ml×kg(-1)×d(-1) respectively. Fluid balance, tissue perfusion, gastrointestinal function recovery time and the imbalance of fluid and electrolyte were recorded. RESULTS: The total fluid input and net fluid balance in restrictive group were significantly fewer than those in control group (total fluid input: 1782.56±258.38 ml/d vs. 2707.50±294.64 ml/d, net fluid balance: 316.67±202.86 ml/d vs. 623.33±244.38 ml/d, both P<0.05), and central venous pressure (CVP) was significantly lower than that in control group (4.03±1.81 mm Hg vs. 6.47±3.09 mm Hg, P<0.05). There were no differences in heart rate (HR) and mean arterial pressure (MAP) between two groups (HR: 85.03±13.49 bpm vs. 81.44±12.49 bpm, MAP: 80.65±11.39 mm Hg vs. 82.38±8.28 mm Hg, both P>0.05). The lactate clearance rate of the first postoperative 24 hours in restrictive group was higher than that in control group [35 (17, 53)% vs. 17 (-6, 33)%, P<0.05]. The times of bowel sounds recovery, the first flatus and stool passed in restrictive group were shorter than those in control group (bowel sounds: 37.43±24.97 hours vs. 46.36±19.34 hours, flatus: 53.63±12.78 hours vs. 75.43±20.07 hours, stool: 78.73±46.48 hours vs. 93.40±41.08 hours, all P<0.05). Vomiting was reduced in the restrictive group compared with control group (2 vs. 7, P<0.05). There were no differences in the occurrences of electrolyte imbalance (5 vs. 3), fluid insufficient (2 vs. 0) and fluid overload (0 vs. 1) between the two groups. CONCLUSION: The postoperative restrictive fluid management by ensuring tissue perfusion can shorten the gastrointestinal function recovery time after elective colonic resection, and may not increase the incidence of water and electrolyte disorders.

Expression and functional role of ALDH1 in cervical carcinoma cells.

Tumor formation and growth is dictated by a very small number of tumor cells, called cancer stem cells, which are capable of self-renewal. The genesis of cancer stem cells and their resistance to conventional chemotherapy and radiotherapy via mechanisms such as multidrug resistance, quiescence, enhanced DNA repair abilities and anti-apoptotic mechanisms, make it imperative to develop methods to identify and use these cells as diagnostic or therapeutic targets. Aldehyde dehydrogenase 1 (ALDH1) is used as a cancer stem cell marker. In this study, we evaluated ALDH1 expression in CaSki, HeLa and SiHa cervical cancer cells using the Aldefluor method to isolate ALDH1-positive cells. We showed that higher ALDH1 expression correlated with significantly higher rates of cell proliferation, microsphere formation and migration. We also could demonstrate that SiHa-ALDH1- positive cells were significantly more tumorigenic compared to SiHa-ALDH1-negative cells. Similarly, SiHa cells overexpressing ALDH1 were significantly more tumorigenic and showed higher rates of cell proliferation and migration compared to SiHa cells where ALDH1 expression was knocked down using a lentivirus vector. Our data suggested that ALDH1 is a marker of cervical cancer stem cells and expand our understanding of its functional role.

A modified triple incision technique for women with locally advanced vulvar cancer: a description of the technique and outcomes.

OBJECTIVE: Women with locally advanced vulvar carcinoma have an excellent chance of a cure by undergoing a radical vulvectomy with an "en bloc" inguinofemoral lymphadenectomy, but the morbidity associated this surgical approach is substantial. To achieve an outcome comparable with the traditional radical method in terms of oncologic safety, and an improved post-operative quality of life, we modified the classic triple-incision technique and suggested it as an alternative for these patients. The aim of this study was to report this new technique. STUDY DESIGN: Between January 2004 and November 2009, 24 patients with clinical stage T2 (≥ 4 cm) or T3 invasive vulvar cancer underwent surgical treatment with our modified triple incision technique. Their clinical and surgical complications and follow-up data were retrospectively reviewed. RESULTS: The post-surgical complications were as follows: lymphoedema in 45.8%, wound breakdown in 20.8% and cellulitis in 8.3%. After a median follow-up of 35.5 months, three (12.5%) patients developed a recurrence in the skin bridge (2/24, 8.3%) or lungs (1/24, 4.2%). All patients suffering from skin bridge recurrences were salvaged by local re-resection. Four (16.7%) cases of death were noted: three (12.5%) patients died of non-cancer-related diseases and one (4.2%) died from a multifocal pulmonary metastasis; no evidence of vulvar or groin disease was observed at these patients' last follow-up. CONCLUSION: The modified triple-incision technique described in this preliminary study appears to be safe, feasible and tolerable for patients with a locally advanced vulvar cancer, and offers an acceptable morbidity.

In-plane orientation effect on the melting behaviour of YBCO thin film.

By means of high-temperature optical microscopy (HTOM), a 60 °C gap in initial melting temperature between two YBa2Cu3O(x) (Y123) thin films was found in situ. Using these two films as seeds, liquid phase epitaxy (LPE) dipping experiments showed the same tendency in the melting behaviour. The in-plane orientation was detected by x-ray diffraction (XRD) pole figure. On the basis of results from HTOM, LPE and XRD, it was unveiled that the interface structure has a predominant influence on the melting mode. A semi-coherent interface suppresses not only the melting growth but also the melting nucleation, while an incoherent interface encourages both of them. (In this work, melting of YBCO refers to the peritectic decomposition of Y123.).