Novel Panel of Long Noncoding RNAs as Diagnostic Biomarkers for Amnestic Mild Cognitive Impairment in Peripheral Blood.

Background: Long noncoding RNAs (lncRNAs) regulate the pathogenesis of Alzheimer's disease (AD). Objective: To identify lncRNAs in the peripheral blood as potential diagnostic biomarkers for amnestic mild cognitive impairment. Methods: In the discovery group, a microarray was used to screen for significant differences in lncRNA expression between patients with mild cognitive impairment (MCI) caused by AD and normal controls (NCs) (n = 10; MCI, 5; NC, 5). Furthermore, two analytic groups were assessed (analytic group 1: n = 10; amnestic MCI (aMCI), 5; NC, 5; analytic group 2: n = 30; AD, 10; aMCI, 10; NC, 10) and finalized in the validation group (n = 150; AD, 50; aMCI, 50; NC, 50). In the analytic and validation groups, real-time quantitative reverse-transcription polymerase chain reaction was used to identify differentially expressed lncRNAs between the aMCI and NC groups. Results: We identified 67 upregulated and 220 downregulated lncRNAs among the expression profiles. The panel with lncRNAs T324988, NR_024049, ENST00000567919, and ENST00000549762 displayed the highest discrimination ability between patients with aMCI and NCs. The area under the receiver operating characteristic curve of this combined model was 0.941, with a sensitivity of 92.00% and specificity of 84.00%. Conclusions: This study reports on a panel of four lncRNAs as promising biomarkers to diagnose aMCIs.

Three-dimensional glass scaffolds improve the In Vitro maturation of porcine cumulus-oocyte complexes and subsequent embryonic development after parthenogenetic activation.

In vitro maturation (IVM) methods for porcine oocytes are still deficient in achieving full developmental capacity, as the currently available oocyte in vitro culture systems still have limitations. In vitro embryo production must also improve the porcine oocyte IVM system to acquire oocytes with good developmental potential. Herein, we tested a three-dimensional (3D) glass scaffold culture system for porcine oocyte maturation. After 42 h, we matured porcine cumulus-oocyte complexes (COCs) on either two-dimensional glass dishes (2D-B), two-dimensional microdrops (2D-W), or 3D glass scaffolds. The 3D glass scaffolds were tested for porcine oocyte maturation and embryonic development. Among these culture methods, the extended morphology of the 3D group maintained a 3D structure better than the 2D-B and 2D-W groups, which had flat COCs that grew close to the bottom of the culture vessel. The COCs of the 3D group had a higher cumulus expansion index and higher first polar body extrusion rate, cleavage rate, and blastocyst rate of parthenogenetic embryos than the 2D-B group. In the 3D group, the cumulus-expansion-related gene HAS2 and anti-apoptotic gene Bcl-2 were significantly upregulated (p < 0.05), while the pro-apoptotic gene Caspase3 was significantly downregulated (p < 0.05). The blastocysts of the 3D group had a higher relative expression of Bcl-2, Oct4, and Nanog than the other two groups (p < 0.05). The 3D group also had a more uniform distribution of mitochondrial membrane potential and mitochondria (p < 0.05), and its cytoplasmic active oxygen species content was much lower than that in the 2D-B group (p < 0.05). These results show that 3D glass scaffolds dramatically increased porcine oocyte maturation and embryonic development after parthenogenetic activation, providing a suitable culture model for porcine oocytes.

Effect of two different laparoscopic techniques on post-operative wound complications in patients with benign gynaecological diseases: A meta-analysis.

Single-port laparoscopy (SPL) has existed for several years. This meta-analysis was conducted to evaluate the efficacy of SPL compared with conventional laparoscopy (CL) in the treatment of benign gynecologic adnexal lesions. The purpose of this meta-analysis is to evaluate the superiority of SPL versus CL in the treatment of post-operative wound pain. The study looked for English-language publications from PubMed, Embase, Cochrane Library and the Web of Science until June 2023. The main result was the visual analogue scale (VAS) after 2, 4, 6, 8, 12, 24 and 48 h after operation. The paper contains 10 related papers by means of e-search. Of these, 4 were randomized controlled trials (RCTs), while 6 were non-RCTs. The results indicated that SPL and CL were significantly different after 2, 24 and 48 h after operation. SPL had lower post-operative pain after 2 h compared with CL (MD, -0.6; 95% CI, -0.98, -0.21; p = 0.002). After the operation, SPL also had a lower incidence of post-operative pain after 24 h compared with CL (MD, -0.59; 95% CI, -1.12, -0.06; p = 0.03). And the difference in pain was at 48 h after the most significant (MD, -0.49; 95% CI, -0.75, -0.23; p = 0.0002). But after 6, 8 and 12 h after operation, there was no significant difference in the degree of pain. Thus, SPL operations may result in a lower degree of pain than CL in both the post-operative and far post-operative phase.

Identification of Key Genes during Ca2+-Induced Genetic Transformation in Escherichia coli by Combining Multi-Omics and Gene Knockout Techniques.

The molecular mechanism of the Ca2+-mediated formation of competent cells in Escherichia coli remains unclear. In this study, transcriptome and proteomics techniques were used to screen genes in response to Ca2+ treatment. A total of 333 differentially expressed genes (317 upregulated and 16 downregulated) and 145 differentially expressed proteins (54 upregulated and 91 downregulated) were obtained. These genes and proteins are mainly enriched in cell membrane components, transmembrane transport, and stress response-related functional terms. Fifteen genes with these functions, including yiaW, ygiZ, and osmB, are speculated to play a key role in the cellular response to Ca2+. Three single-gene deletion strains were constructed with the Red homologous recombination method to verify its function in genetic transformation. The transformation efficiencies of yiaW, ygiZ, and osmB deletion strains for different-size plasmids were significantly increased. None of the three gene deletion strains changed in size, which is one of the main elements of microscopic morphology, but they exhibited different membrane permeabilities and transformation efficiencies. This study demonstrates that Ca2+-mediated competence formation in E. coli is not a simple physicochemical process and may involve the regulation of genes in response to Ca2+. This study lays the foundation for further in-depth analyses of the molecular mechanism of Ca2+-mediated transformation. IMPORTANCE Using transcriptome and proteome techniques and association analysis, we identified several key genes involved in the formation of Ca2+-mediated E. coli DH5α competent cells. We used Red homologous recombination technology to construct three single-gene deletion strains and found that the transformation efficiencies of yiaW, ygiZ, and osmB deletion strains for different-size plasmids were significantly increased. These results proved that the genetic transformation process is not only a physicochemical process but also a reaction process involving multiple genes. These results suggest ways to improve the horizontal gene transfer mechanism of foodborne microorganisms and provide new ideas for ensuring the safety of food preservation and processing.

Novel Plasma miRNAs as Biomarkers and Therapeutic Targets of Alzheimer's Disease at the Prodromal Stage.

BACKGROUND: Amnestic mild cognitive impairment (aMCI) is a prodromal stage of Alzheimer's disease (AD) involving imbalanced beta-site amyloid precursor protein cleaving enzyme 1 (BACE1). MicroRNAs (miRNAs) are associated with AD. OBJECTIVE: This study aimed to investigated whether plasma miRNAs can predict prodromal AD or are associated with AD pathology. METHODS: Participants in the discovery set (n = 10), analysis set (n = 30), and validation set (n = 80) were screened from the China Longitudinal Aging Study. RNA was extracted from the participants' plasma. Microarray sequencing provided miRNA profiles and differentially expressed miRNAs (DEmiRNAs) in the discovery set included patients with 18F-Flutemetamol positron emission tomography scan-confirmed aMCI. Potential biomarkers were screened in the analysis set. The predict capability of candidate miRNAs was assessed in the validation set. Candidate miRNAs modulation of BACE1 expression was explored in rat and human hippocampal neurons in vitro. RESULTS: We verified 46 significant DEmiRNAs between the aMCI and NC groups (p < 0.05), among which 33 were downregulated. In the analysis set, miR-1185-2-3p, miR-1909-3p, miR-22-5p, and miR-134-3p levels decreased significantly in the aMCI group. These miRNAs and previously identified miR-107 were selected as potential biomarkers. A prediction model comprising these five miRNAs showed outstanding accuracy (81.25%) to discriminate aMCI at cut-off value of 0.174. Except for miR-134-3p, the other four miRNAs significantly suppressed Bace1 expression in rat hippocampal neurons in vitro. BACE1 modulation of miR-1185-2-3p, miR-1909-3p, and miR-134-3p was confirmed in human hippocampal neurons in vitro. CONCLUSION: A predictive model consisting of five BACE1-related plasma miRNAs could be a novel biomarker for aMCI.

miR-326 inhibits the cell proliferation and cancer stem cell-like property of cervical cancer in vitro and oncogenesis in vivo via targeting TCF4.

BACKGROUND: Cervical cancer ranks as one of the most prevalent female malignancies globally, and its treatment with new targets has been the focus of current research. The present study set out to investigate the function of microRNA-326 (miR-326) in vitro and in vivo and to verify the direct targeting of transcription factor 4 (TCF4) by miR-326. METHODS: The detection of messenger RNA (mRNA) expressing miR-326 and TCF4 in cervical cancer cell lines and tumor samples was conducted using quantitative real-time polymerase chain (qRT-PCR). A dual-luciferase reporter assay was carried out to detect the target relationship of miR-326 with TCF4. A Cell Counting Kit-8 (CCK-8) assay was employed to detect the effect of miR-326 on CasKi cell viability. Flow cytometry and western blotting were employed to examine the effects of miR-326 on cancer stem cell (CSC)-like property. Tumor weight was measured in orthotopic xenograft mouse models. Immunohistochemistry was employed to analyze the protein expression levels of Ki-67, proliferating cell nuclear antigen (PCNA), CD44, and SRY-box 4 (SOX4). RESULT: Downregulation of the mRNA expression levels of miR-326 was observed in cervical cancer cell lines and tumor tissue, while the levels of TCF4 were upregulated. The dual-luciferase reporter assay revealed binding of miR-326 to the three prime untranslated region (3'-UTR) of TCF4. In vitro assays demonstrated that miR-326 inhibited CasKi cell proliferation through regulating TCF4. miR-326 also suppressed the CSC-like property of CasKi cells by targeting TCF4. Furthermore, the protein expression levels of cyclin D1, ß-catenin, and c-Myc were decreased when miR-326 was added to TCF4-transfected cells. In vivo assays demonstrated that miR-326 inhibited tumor weight, growth, and the protein expression levels of Ki-67, PCNA, CD44, SOX4, and ß-catenin. CONCLUSIONS: miR-326 acted in a tumor-suppressive manner through its regulation of TCF4, and has potential as a biomarker or therapeutic target for cervical cancer.

Magnetic Resonance Imaging of Bilateral Hands Is More Optimal Than MRI of Unilateral Hands for Rheumatoid Arthritis.

OBJECTIVE: To explore the advantages of magnetic resonance imaging (MRI) of bilateral hands in rheumatoid arthritis (RA). METHODS: Consecutive patients with active RA were recruited for clinical assessments, radiographs, and MRI of bilateral hands. Bilateral hands were scanned simultaneously on 3.0 T whole-body MRI system and were scored on synovitis, osteitis, and bone erosion according to the RA MRI scoring (RAMRIS) system. RESULTS: Among 120 patients included, wrist bones and metacarpophalangeal joint (MCPJ) 2 proximal showed bone erosion in early RA. The second to fifth metacarpal bases and the second to fourth MCPJ distal showed more bone erosion in mid-stage or late-stage RA. When MRI of dominant unilateral hand was analyzed, MRI synovitis and osteitis in 5% of wrists and 3 MRI features in 5-14% of MCPJ were misdiagnosed (McNemar test, all p < 0.05). There were 46% wrist synovitis, 29-52% MCPJ2-5 synovitis, 45% wrist osteitis, and 20%-34% MCPJ2-5 osteitis not detected by joint tenderness and/or swelling. When the clinically more severe hand was selected for MRI of unilateral hand according to physical examination, MRI synovitis in 5% of wrists and 3 MRI features in 7-15% of MCPJ were misdiagnosed (all p < 0.05). Scatter plots and linear regression analyses were used to illustrate RAMRIS between dominant or selected hand (Y values) and nondominant or nonselected hand (X values). All linear models were markedly different from a Y = X linear model, indicating the dominant or clinically more severe hand could not represent the contralateral hand to evaluate RAMRIS. CONCLUSION: MRI of bilateral hands is more optimal than MRI of the unilateral hand in RA.

An indirect ELISA for detection of Theileria spp. antibodies using a recombinant protein (rTlSP) from Theileria luwenshuni.

Theileria is a tick-borne, intracellular protozoan parasite of worldwide economic and veterinary importance in small ruminants. Here, an enzyme-linked immunosorbent assay (ELISA) was developed based on Theileria luwenshuni recombinant surface protein (rTlSP) and was used in the standardization and validation of an ELISA for the detection of circulating antibodies against ovine and caprine theileriosis. A total of 233 sera samples were used for the calculation of the cut-off value which served as a threshold between the positive and the negative sera. When the positive threshold was chosen as 19% of the specific mean antibody rate, the specificity was 97.9%, and the sensitivity was 97.1%. There was a cross-reaction with sera against Theileria uilenbergi and Theileria ovis, and no cross-reaction with sera against Babesia spp. in the ELISA and Western blotting. Two hundred forty samples collected from sheep in Gansu province were detected with blood smears and ELISA, respectively. The results showed that the positive rate of Theileria infection in Gansu province were 63.75% with rTlSP-ELISA, and 46.67% with blood smears, respectively. Our test proved that the rTlSP ELISA is suitable to diagnose Theileria infection and could be used in serological surveys to map out the prevalence of ovine and caprine theileriosis.

Circumvention of cisplatin resistance in ovarian cancer by combination of cyclosporin A and low-intensity ultrasound.

Cisplatin resistance is a challenge in the treatment of ovarian cancer. The aim of this study was to explore if ultrasound can overcome chemoresistance and enhance chemosensitization due to cyclosporin A. Ultrasound and/or cyclosporin A were employed to overcome cisplatin resistance in human ovarian cancer cell line COC1/DDP. Mechanisms were explored from the perspective of: DNA damage, intracellular platinum level, detoxification, and genes related to drug efflux and DNA repair. In vivo therapeutic efficacy was validated in a short-term model (subrenal cell-clot transplantation) in mice and the survival benefit was investigated in an orthotopic cancer model in mice using HO-8910PM cells. The findings were: (i) ultrasound enhanced the effect of cisplatin leading to a lower cell-survival rate (IC50 decreased from 3.19 to 0.35 µg/ml); (ii) ultrasound enhanced cisplatin via direct (increasing the intercellular level of active platinum) and indirect (decreasing the glutathione level, and expression of LRP and ERCC1 genes) mechanisms that intensified cisplatin-induced DNA damage, thus enhancing cell apoptosis and necrosis; (iii) cisplatin followed by ultrasound led to small tumor sizes in the short-term model without exacerbation of the systemic toxicity, and prolonged the survival times in the orthotopic model; and (iv) ultrasound synergized the sensitization due to cyclosporin A in vitro and in vivo. These data demonstrated that ultrasound combined with cyclosporin A overcame cisplatin resistance in ovarian cancer.

Detection of DNA damage in sonochemotherapy against cisplatin-resistant human ovarian cancer cells using the modified comet assay.

PURPOSE: To explore methodological modifications in the detection of DNA damage in sonochemotherapy for cisplatin-resistant human ovarian cancer cells using the comet assays. MATERIALS AND METHODS: Chemoresistant cells COC1/DDP were subjected to sonochemotherapy and DNA damage detected with the alkaline and neutral comet assays. RESULTS: In the alkaline assay, the percentage of comets formed was less than that of dead cells, and most values for the percentage of comets formed were < 5% when using the default value to identify comets, showing an underestimation. These values were corrected when adjusting the threshold to the 95th percentile in control cells. In the neutral assay, this modification was not needed. Tail length (TL), tail moment (TM) and Olive tail moment (OTM) dramatically varied between comets. The 75th percentiles of TL, TM and OTM in the alkaline assay, and 90th percentiles in the neutral assay, correlated with the percentage of comets formed, thereby reflecting the temporal shift in DNA damage. Quantification of the interaction using the percentage of comets formed was consistent with that using the percentage of dead cells. CONCLUSIONS: The percentage of comets formed can be used to assess DNA damage in sonochemotherapy against chemoresistant cells when adjusting the threshold.

An epidemiological survey of Theileria infections in small ruminants in central China.

Here, we conducted an epidemiological study in five regions in central China to assess the impact of theileriosis on small ruminants. PCR analysis and microscopic evaluations of blood smears to detect ovine and caprine theileriosis was conducted, in which 256 blood samples and 250 ticks were collected from sheep and goats, and tested for Theileria uilenbergi, T. luwenshuni, and T. ovis. The 18S rRNA gene sequences were deduced from positive samples and used for phylogenetic analysis. The results showed that T. luwenshuni was found most frequently in the five investigated regions and the prevalence of T. luwenshuni was found to be very high by PCR analysis. In contrast, T. uilenbergi and T. ovis infections were not detected in these regions. Phylogenetic tree analysis showed that all of the newly isolated Theileria spp. was in the same clade as T. luwenshuni. Haemaphysalis longicornis, which can transmit T. luwenshuni, was also detected in the sampled sheep and goats in these regions. Our results provide important data to increase the understanding of the epidemiology of ovine and caprine theileriosis, and will aid in the implementation of measures to control theileriosis transmission to small ruminants in central China.

Anticancer potency of cytotoxic drugs after exposure to high-intensity focused ultrasound in the presence of microbubbles and hematoporphyrin.

Chemotherapy is undertaken perioperatively to improve the efficacy of high-intensity focused ultrasound (HIFU) for solid tumors. HIFU at a sufficient intensity for tissue ablation has recently been applied for drug delivery; ultrasonic cavitation plays an important part in HIFU and drug delivery. Hematoporphyrin and microbubbles are adjuncts because they aid cavitation. The effect of HIFU (1.0 MHz; 12,999 W/cm(2) in continuous waves), in the presence of hematoporphyrin and/or microbubbles, on the anticancer potency of 5-fluorouracil, cisplatin, paclitaxel, mitomycin C or adriamycin, was investigated. Insonated adriamycin resulted in a lower death rate of human cancer cells HO-8910 (45.85 ± 2.65% vs 34.84 ± 1.21%, p < 0.05), which was exacerbated when employing hematoporphyrin (34.84 ± 1.21% vs 23.09 ± 7.82%, p < 0.05) or hematoporphyrin combined with microbubbles (34.84 ± 1.21% vs. 8.79 ± 3.69%, p < 0.05); the therapeutic activity was not affected when adding microbubbles alone. High-performance liquid chromatography detected a smaller peak area after subjecting adriamycin to HIFU with the use of hematoporphyrin alone or combined with microbubbles. The other drugs were not affected. Hematoporphyrin, microbubbles and adriamycin increased the throughput of hydroxyl radicals resulting from cavitation as determined by iodine and methylene blue assays. These data suggested that the anticancer activity of a drug may be decreased by HIFU exposure (particularly in the presence of hematoporphyrin and microbubbles). Cavitation produced reactive species that attacked drug molecules, thereby decreasing their antitumor potency; this process was enhanced if the drug itself generated free radicals under insonation.

A rare case of ovarian Burkitt lymphoma associated tumor lysis syndrome.

A 21-year-old woman who presented with pelvic mass, fever and cough was admitted. Ultrasonography revealed a large solid mass and serum CA125 was increased. A bilateral salpingo-oophorectomy was performed and pathological diagnosis showed Burkitt lymphoma of bilateral ovaries. Adjuvant chemotherapy was administrated after surgery. However, on the next day, the patient had an unexpectedly high fever, sigh-like breathing, dilated pupils, and died despite rescue. This is the first report on the post-treatment tumor lysis syndrome with ovarian Burkitt's lymphoma. Identifying patients at risk and initiating therapy early are essential to avoid serious complications associated with tumor lysis syndrome.

Concurrent blockade of the NF-kappaB and Akt pathways potently sensitizes cancer cells to chemotherapeutic-induced cytotoxicity.

Nuclear factor-kappaB (NF-kappaB) and Akt are two major cell survival pathways that are often constitutively activated and can be further stimulated by chemotherpeutics in cancer cells. Although individually targeting the NF-kappaB or Akt has been reported to sensitize caner therapy, the effectiveness of concurrent blocking these two pathways for chemosensitizing of cancer cells to genotoxic therapeutics has not been investigated. In the present study, we investigate the activation of the NF-kappaB and Akt pathways by two frontline anticancer drugs cisplatin and etopside in a variety of cancer cell lines. The effects of blocking these two survival pathways individually or concurrently on cisplatin- or etopside-induced cytotoxicity were detected. The results show that cisplatin and etopside activate both NF-kappaB and Akt in cancer cells. Blockade of either of these pathways with chemical inhibitors or siRNA moderately sensitized cancer cells to cisplatin- or etopside-induced cytotoxicity. Strikingly, much more effective potentiation of cytotoxicity to these anticancer drugs was achieved when NF-kappaB and Akt were concurrently blocked. These data suggest that NF-kappaB and Akt cooperatively attenuate therapeutic-induced cytotoxicity and concurrently blocking these pathways is an effective strategy for improving the anticancer efficacy of therapeutics.

Association of constitutive nuclear factor-kappaB activation with aggressive aspects and poor prognosis in cervical cancer.

INTRODUCTION: Although nuclear factor-kappaB (NF-kappaB) is generally believed to be involved in carcinogenesis, the relationship between NF-kappaB activation and progression of cervical cancer in clinical settings has not been reported. In this study, we investigated the association of NF-kappaB activation with aggressive aspects and prognosis in cervical cancer. METHODS: Nuclear factor-kappaB subunits p65 and p50 were detected in 159 paraffin tissues including normal cervical, precancerous (squamous intraepithelial lesions), and cervical carcinoma tissues by immunohistochemistry. Nuclear factor-kappaB nuclear translocation and DNA-binding activity in precancerous or carcinoma tissues were examined by Western blot and electrophoretic mobility shift assay, respectively. RESULTS: A gradual NF-kappaB activation from normal cervical epithelial cells to precancerous and carcinoma cells was detected by immunohistochemistry (nuclear expression of p65 and p50, P < 0.001), Western blot (NF-kappaB nuclear translocation), and electrophoretic mobility shift assay (enhanced DNA-binding activity). In 79 cancer tissues, increased nuclear p65, an active NF-kappaB form, was correlated with poor tumor grade, lymphatic metastasis, interstitial invasion, and larger tumor size (P < 0.05). Similarly, increased nuclear p50 was correlated with poor tumor grade, interstitial invasion, and larger tumor size (P < 0.05). Moreover, increased nuclear p65 was associated with lower survival rate in patients with cancer (P < 0.05). CONCLUSIONS: Constitutive NF-kappaB activation is correlated to tumor progression, aggressive behaviors, and poor prognosis in cervical cancer, suggesting that NF-kappaB is a tumor promoter, a prognostic indicator, and a possible therapeutic target for this malignant disease.

Correlation of inhibitor of differentiation 1 expression to tumor progression, poor differentiation and aggressive behaviors in cervical carcinoma.

OBJECTIVE: To study the correlation of inhibitor of differentiation 1 (Id-1) to tumor invasion and metastasis by examining Id-1 expression levels in different stages of cervical carcinogenesis. METHODS: Id-1 mRNA and protein expression was detected in total of 171 cervical samples including precancerous and cancerous tissues by quantitative RT-PCR (qRT-PCR) and immunohistochemistry (IHC), respectively. Twenty-five normal cervical tissues were used as a normal control. Correlation between Id-1 positive rates and expression levels to cancer progression and clinicopathologic features was statistically analyzed. RESULTS: A gradual increase of Id-1 protein expression associated with cervical cancer progression was detected (4%, 16%, 50% and 75.9% in normal, low squamous intraepithelial lesion (LSIL), high squamous intraepithelial lesion (HSIL) and cancer tissue, respectively, p<0.001). A similar trend of Id-1 mRNA expression was also observed (1.3, 3.4 and 10.4 fold higher than normal tissues in LSIL, HSIL and cancer tissue, respectively, p<0.001). Furthermore, the Id-1 expression level was correlated to tumor grade (p=0.005), lymph node metastasis (p=0.001), interstitial invasive (p<0.001) and tumor size (p<0.001). These results suggest that high Id-1 expression is associated with tumor growth, invasion and metastasis. CONCLUSION: Id-1 expression is correlated to progression and aggressive behaviors in cervical cancer, suggesting a tumor-promoting role for Id-1 in progression of this malignancy.