Targeted Delivery of Salusin-α Into Rabbit Carotid Arterial Endothelium Using SonoVue.

OBJECTIVES: A new method based on the adhesion of SonoVue to plasmids was assessed to achieve targeted gene delivery into the vascular endothelium. METHODS: pEGFP-Salusin-α and pcDNA3.1-Salusin-α plasmids were transfected into the arterial endothelium of different rabbit groups. Western blotting was performed to analyze the expression of EGFP and salusin-α in the common carotid arteries of rabbits from different groups, and ELISA was performed to detect plasma salusin-α levels in rabbits from each group; simultaneously, blood parameters of different groups of rabbits were measured. RESULTS: Green fluorescence was observed in the right common carotid artery of rabbits transfected with pEGFP-Salusin-α, but not in the endothelial cells of not-transfected control rabbits. The expression of salusin-α in the transfected animals was higher than that in the control not-transfected animals (P < .05). In rabbits transfected with pcDNA3.1-Salusin-α plasmid, salusin-α expression was higher than in the not-transfected control animals (P < .05). However, there was no significant difference in plasma salusin-α levels between transfected animals and controls (P > .05). Blood parameters were also measured in both groups. CONCLUSIONS: Our data confirm the establishment of a new method using SonoVue for targeted gene delivery into the arterial endothelium. Our study outcomes propose a new method of intervention in atherosclerosis and a new tool for targeted gene delivery.

Causal association between blood leukocyte counts and vascular dementia: a two-sample bidirectional Mendelian randomization study.

While previous observational studies have suggested a link between leukocyte counts and vascular dementia (VD), the causal relationship between leukocyte counts and various subtypes of VD remains elusive. This study aimed to investigate the causal relationship between five types of leukocyte counts and VD, with the goal of improving prevention and treatment strategies. In this study, leukocyte counts were used as the exposure variable, with genome-wide association study (GWAS) data sourced from both the UK Biobank and the Blood Cell Consortium. Additionally, GWAS data for five subtypes of vascular dementia were obtained from the FinnGen database. We conducted rigorous statistical analysis and visualization using Mendelian randomization (MR) to elucidate the potential causal relationship between leukocyte counts and vascular dementia. This study, utilizing MR analysis with data from the UK Biobank and Blood Cell Consortium, identified significant causal associations between increased lymphocyte counts and VD. Specifically, lymphocyte counts were found to be causally related to multiple and mixed VD subtypes. Sensitivity analyses, including MR-Egger regression and MR-PRESSO tests, confirmed the robustness of these findings, with no evidence of reverse causality or significant horizontal pleiotropy detected. The results underscore a potential inflammatory or immunological mechanism in the pathogenesis of VD, highlighting lymphocytes as a key component in their etiology. This investigation establishes a robust association between elevated lymphocyte and leukocyte counts and an increased risk of VD, emphasizing the roles of inflammation, immune activation, and hematological factors in disease pathogenesis.

Ancient genomes illuminate the demographic history of Shandong over the past two millennia.

Shandong province, located in the Lower Yellow River, is one of the birthplaces of ancient Chinese civilization. However, the comprehensive genetic histories of this region have remained largely unknown until now due to a lack of ancient human genomes. Here, we present 21 ancient genomes from Shandong dating from the Warring States period to the Jin-Yuan Dynasties. Unlike the early Neolithic samples from Shandong, the historical samples are most closely related to post-Late Neolithic populations of the Middle Yellow River Basin, suggesting a population turnover in Shandong from the Neolithic Age to the Historical era. In addition, we detect a close genetic affinity between the historical samples in Shandong and present-day Han Chinese, showing long-term genetic stability in Han Chinese at least since the Warring States period.

Animals in Mortuary Practices of Bronze-Age Pastoral Societies: Caprine Use at the Site of Dunping in Northwestern China.

The late second and first millennium BC witnessed extensive economic, cultural, and political exchanges between pastoralists and sedentary farming states in East Asia. Decades of archaeological fieldwork across northern China have revealed a large number of burial sites associated with pastoralists during the first millennium BC. These sites were characterized by the inhumation of specific animal parts in burials, predominantly the skulls and hooves of sheep, goats, cattle, and horses. However, the selection preference for these animals and how they were integrated into the mortuary contexts of these pastoral societies remain poorly investigated. Here, we report a preliminary analysis of caprine remains from 70 burials at the site of Dunping in the southern Gansu region of northwestern China, dated to approximately the seventh to fourth centuries BC. Based on an examination of species composition, post-depositional effects, traces of human alteration, skeletal element representation, and age at death, we discussed the selection, slaughtering, and inhumation of caprines concerning the mortuary practices at the site. Comparisons between Dunping and several other contemporaneous burial sites in neighboring regions, specifically in terms of the mortality profiles, further highlight distinct patterns in the selection of caprines for mortuary purposes among pastoral societies. These differences suggest varying degrees of emphasis placed on the economic and social significance attributed to caprines. Our findings provide new insights into the roles that caprines played in both ritual performances and subsistence practices among pastoralists in East Asia during the first millennium BC.

Construction and Verification of a Combined Hypoxia and Immune Index for Clear Cell Renal Cell Carcinoma.

Clear cell renal cell carcinoma (ccRCC) is one of the most aggressive malignancies in humans. Hypoxia-related genes are now recognized as a reflection of poor prognosis in cancer patients with cancer. Meanwhile, immune-related genes play an important role in the occurrence and progression of ccRCC. Nevertheless, reliable prognostic indicators based on hypoxia and immune status have not been well established in ccRCC. The aims of this study were to develop a new gene signature model using bioinformatics and open databases and to validate its prognostic value in ccRCC. The data used for the model structure can be accessed from The Cancer Genome Atlas database. Univariate, least absolute shrinkage and selection operator (LASSO), and multivariate Cox regression analyses were used to identify the hypoxia- and immune-related genes associated with prognostic risk, which were used to develop a characteristic model of prognostic risk. Kaplan-Meier and receiver-operating characteristic curve analyses were performed as well as independent prognostic factor analyses and correlation analyses of clinical characteristics in both the training and validation cohorts. In addition, differences in tumor immune cell infiltrates were compared between the high and low risk groups. Overall, 30 hypoxia- and immune-related genes were identified, and five hypoxia- and immune-related genes (EPO, PLAUR, TEK, TGFA, TGFB1) were ultimately selected. Survival analysis showed that the high-risk score on the hypoxia- and immune-related gene signature was significantly associated with adverse survival outcomes. Furthermore, clinical ccRCC samples from our medical center were used to validate the differential expression of the five genes in tumor tissue compared to normal tissue through quantitative real-time polymerase chain reaction (qRT-PCR). However, more clinical trials are needed to confirm these results, and future experimental studies must verify the potential mechanism behind the predictive value of the hypoxia- and immune-related gene signature.

Optimization of whole-cell vaccines with CpG/αOX40/cGAMP to strengthen the anti-tumor response of CD4+ T cells in melanomas.

METHODS: In this study, we developed a strategy for the prevention and therapy of melanoma using a whole-cell vaccine combined with a CpG/αOX40/cGAMP triple adjuvant. The CpG/αOX40/cGAMP triple adjuvant was used to co-culture melanoma cells in vitro to induce immunogenic death of tumor cells. The mixture of inactivated tumor cells and the triple drug was an optimized tumor whole-cell vaccine, which was injected subcutaneously into mice for tumor prevention and therapy. Furthermore, we analyzed the changes of immune cells in spleen and tumor by flow cytometry and immunohistochemistry, and detected the changes of cytokines after vaccine application by cytometric bead array to explore the specific mechanism of vaccine. RESULTS: In vaccine prevention and therapy experiments, it was observed that the tumor growth was significantly inhibited in the whole-cell vaccine group, and the survival time of mice was significantly prolonged. Flow cytometry results showed that the proportion of CD4+ T cells and CD8+ T cells in tumor of mice in vaccine group was higher than that in control group, especially the CD4+ T cells. CONCLUSION: The optimized vaccine has the unique ability to amplify tumor-specific CD4+ T cells, which improves antitumor sensitivity, and has a significant effect on the prevention and therapy of melanoma mice.

Detection of IgG anti-A/B must be essential for safe transfusion support in patients undergoing ABO incompatible allogeneic HSCT.

Recipients of ABO incompatible allogeneic HSCT present with unusual ABO groups and crossmatch problems accompanied by change of the ABO group from that of the host to the donor. We report on a patient undergoing major ABO incompatible allogeneic HSCT (donor/recipient: A/O) whose blood group was wrongly established to have completely switched to blood group A when using a routine tube method and micro gel column blood grouping card on the 68th day post transplantation. The major crossmatch test with added antiglobulin and blood group A red cells was still positive. After further investigation, an explanation was found because we could not detect IgG anti-A in the serum. At the same time, anti-A coated the patient's RBCs and could be identified using a heat elution method although the DAT was negative. We also found an obvious mixed field with the LISS-IAT gel card. Hence, routine methods of ABO grouping are unfit for ABO incompatible allogeneic HSCT patients and a micro column neutral gel card is recommended for forward typing especially to detect mixed fields and a LISS-IAT gel card or IAT tube method for detecting IgG anti-A/B in reverse typing.

The investigation of platelet transfusion refractory in 69 malignant patients undergoing hematopoietic stem cell transplantation.

BACKGROUND: Platelet transfusion refractoriness (PTR) is thought to be associated with HPA and HLA incompatibility but be unrelated to ABO incompatibility in practice. In this study we aim to investigate the incidence of PTR and potential risks in patients undergoing hematopoietic stem cell transplantation (HSCT). METHOD: A total of 69 HSCT patients were involved with their basic characteristics recorded. PTR was identified by 24h corrected count increment (24h-CCI) post platelet transfusion and we transformed it to the corrected platelet increment (CPI), which was compared between the PTR and non PTR groups. Age, gender, ABO incompatibility, disease and CPI were analyzed by Logistic regression analysis for PTR incidence. We searched our medical records to find possible risks of PTR with different levels of CPI. RESULTS: There was no significant difference of platelet engraftment (PE) (P=0.271) and platelet requirement (PR) (P=0.333) between patients with ABO matched and mismatched transplants. Thirteen patients experienced PTR but with a varied refractory ratio (20-95%). Logistic regression analysis showed that the CPI <10×10(9)/l (P=0.006) was dramatically related to the PTR while ABO incompatibility (P=0.319), MDS (P=0.552), PLT count pre-transplantation <50×10(9)/l (P=0.998) were of no statistical significance. There were 142U (32.7%) of platelets transfused with unsatisfactory CPIs, mainly (48.6%) within the second week since transplantation. From the investigation of medical records, infections, fever and bleeding were the most common reasons for PTR. CONCLUSIONS: PTR is a common phenomenon but more associated with non-immune causes in HSCT. The quantification of CPI may imply potential risks of PTR and help clinicians to better use platelet apheresis.

Everolimus vs. rapamycin for treating diabetic nephropathy in diabetic mouse model.

In order to evaluate the effectiveness of everolimus vs. rapamycin in the treatment of diabetic nephropathy, 8-week old diabetic (db/db) mice received everolimus (2 mg/kg every day) or rapamycin (2 mg/kg every day) for 4 weeks or 12 weeks respectively. Blood and 24-h urine samples were collected for biochemical tests. One kidney from each mouse was homogenized for protein analysis and the other was removed for histological analysis. The expression levels of transforming growth factor-ß1 (TGF-ß1)and phospho-p70s6k were detected by using ELISA and Western blot, respectively in the renal tissue as well as in mesengial cell culture samples. Everolimus was significantly more effective than rapamycin in improving indexes of renal function and glomerular hypertrophy, and in decreasing accumulation and expansion of the extracellular matrix. However, everolimus inhibited TGF-ß1 secretion and p70s6k phosphorylation induced by high glucose in vitro less efficiently than rapamycin at the same dose. Everolimus was more effective than rapamycin in preventing diabetic nephropathy in vivo, which may be contributed to the fact that everolimus has better bioavailability and a higher oral absorption rate.

Predictive Risk Factors for Early Recurrence of Stage pIIIA-N2 Non-Small Cell Lung Cancer.

PURPOSE: Inflammatory biomarkers and clinical pathological factors have been reported to predict survival of patients with non-small cell lung cancer (NSCLC). The goal of this study was to identify risk factors for early recurrence in patients with pIIIA-N2 NSCLC who had undergone radial resection. METHODS: A retrospective analysis was conducted on 238 patients with pIIIA-N2 NSCLC who underwent surgical treatment at the First Affiliated Hospital of Wenzhou Medical University between December 2006 and August 2018. The early recurrence (ER) group included patients who recurred within one year of curative resection, while the non-early recurrence (NER) group included patients who did not recurrence or recurrence beyond one year. The univariate and multivariate Cox proportional risk analyses were used to identify prognostic factors associated with early recurrence, while the chi-square test was used for categorical data. Overall survival and recurrence-free survival were assessed by Kaplan-Meier estimates. RESULTS: A total of 69 patients experienced an early recurrence, while the remaining 169 patients did not relapse within one year. ER patients had a much worse prognosis than NER patients, with median survival times of 20.6 and 83.1 months, respectively. Multivariate analysis showed that smoking status, tumor size, metastatic lymph node ratio (LNR) and platelet-to-lymphocyte ratio (PLR) were independent risk factor of early recurrence. Patients with early recurrence were more likely to develop bone metastases. CONCLUSION: Smoking history, large tumour size, and elevated LNR and PLR values in pIIIA-N2 NSCLC patients after complete resection may have a significant risk of early recurrence. Based on these independent risk indicators, this prediction model may successfully predict early recurrence and advise individual treatment.

PCA3 gene expression in prostate cancer tissue in a Chinese population: quantification by real-time FQ-RT-PCR based on exon 3 of PCA3.

Prostate cancer (PCa) is the second most common cancer in men, and its incidence is still increasing. PCA3 is the most prostate cancer specific biomarker. Here we confirmed that both exon 3 and exon 4 are in the prostate-specific region of the PCA3 gene, and established the methodology of real-time fluorescent quantitative RT-PCR (FQ-RT-PCR) detecting PCA3 mRNA with primer spanning exons 1 and 3, and evaluated its clinical utility in a Chinese population. What disclosed that PCA3 mRNA is prostate cancer specific and shows increased expression in prostate cancer. It could be a reliable molecular marker in prostate cancer diagnosis. Exon 3-based real-time FQ-RT-PCR may prove useful in prostate cancer diagnosis, given that the associated primer would span only exons 1 and 3, relative to other models spanning exons 1 to 4. A shorter amplicon would not only enhance the efficiency of real-time FQ-RT-PCR, but may also simplify the quantification of PCA3 mRNA.

miR-205-5p Mediated Downregulation of PTEN Contributes to Cisplatin Resistance in C13K Human Ovarian Cancer Cells.

Cisplatin resistance is a major cause of treatment failure in advanced ovarian cancer. The limited evidence shows the paradoxical regulation of miR-205 on chemotherapy resistance in cancer. Herein, we found that miR-205-5p was enormously increased in cisplatin-resistant C13K ovarian cancer cells compared with its cisplatin-sensitive OV2008 parental cells using miRNA microarrays, which was further verified by quantitative PCR. Furthermore, we confirmed that inhibition of miR-205-5p upregulated PTEN and subsequently attenuated its downstream target p-AKT, which inversed C13K cells from cisplatin resistance to sensitivity. Our data suggest that miR-205-5p contributes to cisplatin resistance in C13K ovarian cancer cells may via targeting PTEN/AKT pathway.

[Quantitative detection of DD3 mRNA in prostate cancer tissues by real-time fluorescent quantitative reverse transcription polymerase chain reaction].

OBJECTIVE: To analyze the expression of DD3 mRNA in the prostate tissues. METHODS: DD3 mRNA was detected by realtime fluorescent quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR) based on the Taqman technique in the tissues of 27 patients with non-prostate cancer( NPCa), 21 prostate cancer( PCa), 39 benign prostatic hyperplasia (BPH) and 15 normal prostate (NP). The ROC curve was used to evaluate the diagnostic value of DD3 mRNA. RESULTS: DD3 mRNA expression was not detected in the NPCa tissues. The median expressions of DD3 mRNA in PCa, BPH and NP tissues were 7. 2 x 10(6), 2. 5 x 10(4) and 1.5 x 10(4) copies/mg tissue, respectively. The DD3 mRNA expression levels were significantly different between nonmalignant and malignant tissues (P < 0.01). No significant differences in DD3 mRNA expression were detected between the NP and BPH tissues and no significant correlation was found between the DD3 mRNA expression and clinical pathological parameters. The AUC-ROC was 0.937 (95% CI: 0.879 - 0.995) at cutoff value 1.4 x 10(5) copies/mg tissue. The sensitivity, specificity, accuracy, positive predictive value, negative predictive value, positive likelihood ratio and negative likelihood ratio for DD3 were 90.5%, 85.0%, 86.7%, 76.0%, 94.3%, 6.03 and 0.11 respectively. CONCLUSION: The DD3 mRNA expression is confined to prostate tissues and highly upregulated in PCa tissues. It has a potential application value in the early diagnosis of prostate cancer and the follow-up of the patient.

[Quantitative detection of differential display code 3 mRNA in peripheral blood of patients with prostate cancer].

OBJECTIVE: To investigate the expression of DD3 mRNA in the peripheral blood and its value in diagnostic of prostate cancer (Pca). METHODS: Thirty-five untreated Pca patients, aged 72 (53 - 83), 58 Pca patients treated with endocrinotherapy, aged 74 (53 - 86), and 59 patients with benign prostatic hyperplasia, aged 71 (48 - 85), underwent peripheral blood sample collection one week before or after digital examination. RT-PCR was used to examine the level of DD3 mRNA. The level of prostate specific antigen (PSA) was detected too. Ten healthy male frontiers, aged 33 (21 - 38), were used as controls. Receiver operating curve (ROC) was drawn to evaluate the diagnostic performance of DD3 mRNA. RESULTS: The DD3 mRNA level of the untreated Pca patients was 2741 copies/ml, significantly higher than those of the Pca patients treated with endocrinotherapy, patients with benign hyperplasia, and healthy persons (all < 24 copies, all P < 0.001). The DD3 mRNA level of the endocrinotherapy-treated Pca patients was significantly higher than those of the patients with benign hyperplasia, and healthy persons (both P < 0.001). ROC analysis showed that when the critical value was 846 copies/ml the area under curve of ROC (AUC-ROC) was 0.8233 (95% CI: 0.725 - 0.910). and the sensitivity was 74.34%, the specificity was 89.8%. The DD3 mRNA in the peripheral blood increased along with the increase of clinical staging (P < 0.01). CONCLUSION: The level of DD3 mRNA in the peripheral blood is an excellent marker for diagnosis pf Pca, and may help in monitoring of the curative effects of endocrinotherapy.

Stepwise discriminant function analysis for rapid identification of acute promyelocytic leukemia from acute myeloid leukemia with multiparameter flow cytometry.

Diagnosis of acute promyelocytic leukemia (APL) has been accelerated by multiparameter flow cytometry (MFC). However, diagnostic interpretation of MFC readouts for APL depends on individual experience and knowledge, which inevitably increases the risk of arbitrariness. We appraised the feasibility of using stepwise discriminant function analysis (SDFA) based on MFC to optimize the minimal variables needed to distinguish APL from other acute myeloid leukemia (AML) without complicated data interpretation. Samples from 327 patients with APL (n = 51) and non-APL AML (n = 276) were randomly allocated into training (243 AML) and test sets (84 AML) for SDFA. The discriminant functions from SDFA were examined by correct classification, and the final variables were validated by differential expression. Finally, additional 20 samples from patients with atypical APL and AML confusable with APL were also identified by SDFA method and morphological analysis. The weighed discriminant function reveals seven differentially expressed variables (CD2/CD9/CD11b/CD13/CD34/HLA-DR/CD117), which predict a molecular result for APL characterization with an accuracy that approaches 99% (99.6 and 98.8% for AML samples in training and test sets, respectively). Furthermore, the SDFA outperformed either single variable analysis or the more limited 3-component analysis (CD34/CD117/HLA-DR) via separate SDFA, and was also superior to morphological analysis in terms of diagnostic efficacy. The established SDFA based on MFC with seven variables can precisely and rapidly differentiate APL and non-APL AML, which may contribute to the urgent initiation of all-trans-retinoic acid-based APL therapy.

Diagnosis of Drug Resistance to Fluoroquinolones, Amikacin, Capreomycin, Kanamycin and Ethambutol with Genotype MTBDRsl Assay: a Meta-Analysis.

BACKGROUND: The Genotype MTBDRsl is a new-generation PCR-based line-probe assay for rapid identification of the resistance to the second-line antituberculosis drugs with a single strip. The aim of this meta-analysis was to evaluate the performance of Genotype MTBDRsl in detecting drug resistance to fluoroquinolones, amikacin, capreomycin, kanamycin and ethambutol in comparison with the phenotypic drug susceptibility test. DESIGN: We searched Pubmed, Embase and the Cochrane Library and calculated the sensitivity, the specificity, the positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR), corresponding 95% confidence interval (CI), and the area under the summary receiver operating characteristic (SROC) curves (AUC), and tested heterogeneity in accuracy estimates with the Spearman correlation coefficient and Chi-square. RESULTS: The summarized sensitivity (95% CI), specificity (95% CI), and AUC (standard error) were 0.869 (0.847-0.890), 0.973 (0.965-0.979) and 0.9690 (0.0188) for fluoroquinolones, 0.868 (0.829-0.900), 0.998 (0.994-0.999) and 0.9944 (0.0050) for amikacin, 0.879 (0.838-0.914), 0.970 (0.958-0.978) and 0.9791 (0.0120) for capreomycin, 0.501 (0.461-0.541), 0.991 (0.983-0.996) and 0.9814 (0.0114) for kanamycin and 0.686 (0.663-0.709), 0.871 (0.852-0.888) and 0.7349 (0.0639) for ethambutol, respectively. CONCLUSION: The genotype MTBDRsl demonstrate excellent accuracy for detecting drug resistance to fluoroquinolones, amikacin, and capreomycin, but it may not be an appropriate choice for detection of kanamycin and ethambutol.

IL-1ß+3953C/T, -511T/C and IL-6 -174C/G polymorphisms in association with tuberculosis susceptibility: A meta-analysis.

BACKGROUND: Studies of the association between the interleukin-1ß gene (IL-1ß) (+3953C/T, -511T/C) and interleukin-6 gene (IL-6) (-174G/C) polymorphisms and susceptibility to tuberculosis (TB) have yielded inconsistent results. The aim of this study was to investigate the relationship between these polymorphisms and TB risk by this meta-analysis. METHODS: We systematically searched published literatures on IL-1ß gene and IL-6 gene polymorphisms and tuberculosis risk by the PubMed, Medline, Embase, Web of Science, Elsevier Science Direct and Cochrane Library databases, and identified outcome data from all articles. Statistical analysis was performed using Stata 12.0. A total of 15 studies comprising 3983TB patients and 3996 controls were included in the present study. RESULTS: We found that the IL-6 -174C/G polymorphism might be associated with decreased risk of TB (C vs. G: OR=0.77, 95% CI=0.64-0.91; CG vs. GG: OR=0.72, 95% CI=0.57-0.90; CC+CG vs. GG: OR=0.71, 95% CI=0.57-0.88). In the stratified analysis by ethnicity, significantly decreased risk were observed for IL-1ß -511T/C polymorphism in Africans (C allele vs. T allele: OR=0.86, 95% CI=0.74-0.99; CC vs. TT: OR=0.74, 95% CI=0.55-0.99). Moreover, the IL-6 -174 C/G polymorphism was also associated with decreased risk of TB in Asians (C vs. G: OR=0.71, 95% CI=0.54-0.93; CG vs. GG: OR=0.61, 95% CI=0.44-0.85; CC+CG vs. GG: OR=0.63, 95% CI=0.46-0.86). We also performed the analyses by sample types in IL-6 -174 C/G polymorphism, and significant decreased TB risk was observed in pulmonary tuberculosis group (C allele vs. G allele: OR=0.69, 95% CI=0.52-0.93; CG vs. GG: OR=0.69, 95% CI=0.52-0.91; CC+CG vs. GG: OR=0.71, 95% CI=0.55-0.93), and pulmonary tuberculosis plus extra-pulmonary tuberculosis mixed group (CC vs. GG: OR=0.44, 95% CI=0.22-0.88; CC vs. CG+GG: OR=0.48, 95% CI=0.25-0.94). CONCLUSIONS: This meta-analysis indicates that the IL-1ß -511T/C polymorphism is associated with TB decreased risk in Africans, and IL-6 -174C/G polymorphism in Asians. Further well-designed, large scale studies are required to confirm this conclusion.

microRNA-218 expression and its association with the clinicopathological characteristics of patients with cervical cancer.

The aim of the present study was to investigate the expression of microRNA-218 (miRNA-218) in the serum and cervical tissue and its association with the clinicopathological features of cervical cancer (CC). The expression of miRNA-218 was detected in the serum and cervical tissue of 112 patients with CC and 50 age-matched hysteromyoma patients via the reverse transcription-quantitative polymerase chain reaction. The clinical data were collected and the association between the expression of miRNA-218 and the clinicopathological characteristics of the patients was analyzed. The expression of miRNA-218 in the cancer group was significantly decreased in the cervical tissue and serum compared with that in the control group (P<0.001). The decreased expression of miRNA-218 was associated with a later International Federation of Gynecology and Obstetrics stage, a more invasive pathological type and lymphatic node metastasis but not with age, age at menarche, menopausal status, number of pregnancies and deliveries, family history of cancer or tumor size. In conclusion, miRNA-218 was found to be downregulated in the cancer tissue and serum of the patients with CC. The decreased expression of miRNA-218 in CC was associated with the invasiveness of the tumor.

ABO chimerism determined by real-time polymerase chain reaction analysis after ABO-incompatible haematopoietic stem cell transplantation.

BACKGROUND: The dynamic alteration of ABO blood group in ABO-incompatible haematopoietic stem cell transplantation can be well defined by ABO chimerism analysis. In view of the intrinsic difference in ABO phenotypes or genotypes between donor and recipient in ABO incompatible transplantation, ABO allele-associated nucleotide polymorphic sites could theoretically be used as available informative markers for chimerism analysis. MATERIALS AND METHODS: We chose nucleotide polymorphism sites (261, 467, 802, 803 and 1,061) of common ABO alleles to use as markers from 76 randomly chosen donors and assessed the limit of linear detection of a specifically designed real-time quantitative polymerase chain reaction using SYBR Green I dye with these sites for a chimerism assay. RESULTS: We successfully established the real-time quantitative polymerase chain reaction for detecting 261, 467(mutated) and 803 sites and their limits of linear detection, which were at least 10(-3), 10(-2) and 10(-2), respectively. The limits of linear detection between heterozygous DNA and homozygous DNA with 261 or 803 sites were not significantly different. DISCUSSION: ABO chimerism can be well analysed by real-time polymerase chain reaction with ABO gene-associated nucleotide polymorphic sites. This strategy could be very beneficial for the early and accurate judgement of the crucial point of transition in order to plan a safe transfusion strategy following ABO-incompatible transplantation.