Detection of pulmonary actinomycosis by metagenomic next-generation sequencing in a cancer patient receiving maintenance olaparib: A review and case report.

A 52-year-old female patient receiving olaparib maintenance treatment was admitted to hospital with a low fever and chest tightness. A CT scan of the patient's chest showed diffuse ground glass shadow or miliary nodular shadow in the bilateral lungs. Her inflammatory biomarkers were almost normal, except the slightly elevated C-reactive protein. Moreover, lymphocytes count obviously decreased. Empirical treatment did not relieve her symptoms, while traditional testing developed negative results. The results of metagenomic next-generation sequencing (mNGS) revealed the presence of a potential pathogen, Actinomyces odontolyticus (A. odontolyticus), in bronchoalveolar lavage fluid (BLAF). Once large-dosed penicillin G was administered, the fever returned to normal and chest tightness disappeared. Reexamination of chest CT revealed that the pulmonary lesions was almost absorbed. Our case demonstrated that mNGS is a novel approach to identify pathogens sensitively and accurately, especially for uncommon and atypical infection.

Mechanism of Hirudin-Mediated Inhibition of Proliferation in Ovarian Cancer Cells.

To investigate the inhibitory effect of hirudin on the cell proliferation of human ovarian cancer A2780 cells by preventing thrombin and its underlying molecular mechanism. Cell Counting Kit-8 (CCK-8) method was used to detect the effect of different concentrations of hirudin and thrombin on the cell proliferation of A2780 cells. PAR-1 wild-type overexpression plasmid was constructed utilizing enzyme digestion identification, and it was transferred to A2780 cells. Sequencing and Western blot were used to detect the changes in PAR-1 protein expression. Western blot detection of PKCα protein phosphorylation in A2780 cells was performed. We also implemented quantitative PCR to detect the mRNA expression levels of epithelial-mesenchymal transition (EMT)-related genes, CDH2, Snail, and Vimentin, in A2780 cells. 1 µg/ml hirudin treatment maximally inhibited the promotion of A2780 cell proliferation by thrombin. Hirudin inhibited the binding of thrombin to the N-terminus of PAR-1, hindered PKCα protein phosphorylation in A2780 cells, and downregulated the mRNA expression levels of CDH2, Snail, and Vimentin. In conclusion, hirudin inhibits the cell proliferation of ovarian cancer A2780 cells, and the underlying mechanism may be through downregulating the transcription level of EMT genes, CDH2, Snail, and Vimentin. This study indicates that hirudin may have a therapeutic potential as an anti-cancer agent for ovarian cancer.

Systemic immune-inflammation index predicts prognosis and responsiveness to immunotherapy in cancer patients: a systematic review and meta­analysis.

The systemic immune-inflammation index (SII) is a significant prognostic factor in some cancer types. However, the prognostic role of SII in cancer patients with immunotherapy remains uncertain. We aimed to evaluate the relationship between pretreatment SII and clinical survival outcomes for advanced-stage cancer patients treated with immune checkpoint inhibitors (ICIs). A comprehensive literature search was performed to identify eligible studies concerning the association between pretreatment SII and survival outcomes in advanced cancer patients treated with ICIs. The data were extracted from publications and used to calculate the pooled odds ratio (pOR) for objective response rate (ORR), disease control rate (DCR), and pooled hazard ratio (pHR) for overall survival (OS), progressive-free survival (PFS), along with 95% confidence intervals (95% CIs). Fifteen articles with 2438 participants were included. A higher level of SII indicated a lower ORR (pOR = 0.73, 95% CI 0.56-0.94) and worse DCR (pOR = 0.56, 95% CI 0.35-0.88). High SII was associated with a shorter OS (pHR = 2.33, 95% CI 2.02-2.69) and unfavorable PFS (pHR = 1.85, 95% CI 1.61-2.14). Therefore, high SII level might be a non-invasive and efficacious biomarker of poor tumor response and adverse prognosis of advanced cancer patients with immunotherapy.

Reaction Analysis and Process Optimization with Online Infrared Data Based on Kinetic Modeling and Partial Least Squares Quantitation.

In-situ Fourier transform infrared (FT-IR) spectroscopy has been recognized as an important technology for online monitoring of chemical reactions. However, analysis of the real-time IR data for identification and quantification of uncertain reactants or intermediates is often ambiguous and difficult. Here, we propose an analysis algorithm based on reaction kinetic modeling and the chemometric method of partial least squares (PLS) to comprehensively and quantitatively study reaction processes. Concentration profiles and apparent kinetic parameters can be simultaneously calculated from the spectral data, without the demand of complicated analysis on characteristic absorbance peaks or tedious sampling efforts for multivariate modeling. Paal-Knorr reactions and glyoxylic acid synthesis reactions were selected as typical reactions to validate the algorithm. A lack of fit of the Paal-Knorr reaction spectra was less than 2.5% at various conditions, and the absolute errors between the predicted values and HPLC measurement of glyoxylic acid synthesis were less than 6% during the reaction process. Moreover, the reaction kinetic models extracted from FT-IR data were used to simulate reaction processes and optimize the conditions in order to maximize product yields, which proved that this analysis method could be used for process optimization.

Prognostic Nutritional Index Predicts Response and Prognosis in Cancer Patients Treated With Immune Checkpoint Inhibitors: A Systematic Review and Meta-Analysis.

Objective: To investigate the association between pretreatment prognostic nutritional index (PNI) and clinical survival outcomes for advanced-stage cancer patients treated with immune checkpoint inhibitors (ICIs). Methods: We conducted a comprehensive literature search to identify eligible studies concerning the relationship between pretreatment PNI and survival outcomes in advanced cancer patients treated with ICIs. Published data were extracted and pooled odds ratio (pOR) for objective response rate (ORR), disease control rate (DCR), and pooled hazard ratio (pHR) for overall survival (OS), progressive-free survival (PFS), along with 95% confidence intervals (95% CIs) were calculated. Results: Twelve studies with 1,359 participants were included in our study. A higher level of PNI indicated a greater ORR (pOR = 2.17, 95% CI = 1.52-3.10) and favorable DCR (pOR = 2.48, 95% CI = 1.87-3.29). Low PNI was associated with a shorter OS (pHR = 2.24, 95% CI = 1.57-3.20) and unfavorable PFS (pHR = 1.61, 95% CI = 1.37-1.88). Conclusion: Low PNI might be an effective biomarker of poor tumor response and adverse prognosis of advanced cancer patients with ICIs. Further studies are needed to verify the prognostic value of PNI in clinical practice.

Mechanism and kinetic study of Paal-Knorr reaction based on in-situ MIR monitoring.

An in-depth understanding of reaction processes is beneficial to the development and quality control of chemical products. In this work, the mechanism and kinetics of the Paal-Knorr reaction for pyrrole derivatives are thoroughly studied using in-situ Fourier transform infrared (FTIR) spectroscopy. The hemiacetal amine intermediate, reactants, and products were identified and quantified by the treatment of real-time infrared spectra via chemometrics method and two-dimensional correlation spectroscopy (2DCOS) technique. Based on the IR quantitative models, influences of operating conditions on reaction processes were investigated, and the reaction kinetic model was built with kinetic parameters of two rate-limiting reaction steps calculated. This approach of analysis on the in-situ FTIR data demonstrated the ability to extract useful information on reaction components, especially the intermediate spectrum, from the confounding real-time IR data. The in-situ FTIR monitoring combined with the IR analysis methods is proved as a powerful tool for revealing the reaction mechanism and kinetics.

Prognostic value of platelet-to-lymphocyte ratio in neoadjuvant chemotherapy for solid tumors: A PRISMA-compliant meta-analysis.

INTRODUCTION: Previous research indicates that the platelet-to-lymphocyte ratio (PLR) may be an indicator of poor prognosis in many tumor types. However, the PLR is rarely described in patients undergoing neoadjuvant chemotherapy (NAC) for solid tumors. Thus, we performed a meta-analysis to investigate the prognostic value of this ratio for patients with solid tumors treated by NAC. METHODS: A comprehensive search of the literature was conducted using the PubMed, EMBASE, Cochrane Library, and Web of Science databases, followed by a manual search of references from the retrieved articles. Pooled hazard ratios (HRs) with 95% confidence interval (CIs) were used to evaluate the association between PLR and 3 outcomes, namely, overall survival, disease-free survival, and pathological complete response rate after NAC. RESULTS: Eighteen studies published no earlier than 2014 were included in our study. A lower PLR was associated with better overall survival (HR = 1.46, 95% CI, 1.11-1.92) and favorable disease-free survival (HR = 1.81, 95% CI, 1.27-2.59). A PLR that was higher than a certain cutoff was associated with a lower pathological complete response rate in patients with cancer who received NAC (Odds ratio = 1.93, 95% CI, 1.40-2.87). CONCLUSION: Elevated PLR is associated with poor prognosis in various solid tumors. PLR may be a useful biomarker in delineating those patients with poorer prognoses who may benefit from neoadjuvant therapies.

MiR-221-3p-mediated downregulation of MDM2 reverses the paclitaxel resistance of non-small cell lung cancer in vitro and in vivo.

MicroRNAs (miRNAs) are involved in the initiation and development of cancer and participate in drug resistance. Paclitaxel (PTX) is a first-line chemotherapy drug for advanced non-small cell lung cancer (NSCLC). The abnormal miRNA expression in NSCLC and its association with chemotherapy drug resistance remains largely unknown. The study aimed to investigate the aberrant expression of miR-221-3p in NSCLC and to elucidate its molecular mechanisms in relation to PTX resistance. PTX increased miR-221-3p expression and regulated MDM2/P53 expression in the PTX-sensitive NSCLC strain (A549). Meanwhile, miR-221-3p was rarely expressed and not interfered by PTX in PTX-resistant A549 cells (A549/Taxol). Dual-luciferase reporter assay confirmed that miR-221-3p specifically binds to MDM2 messenger RNA and inhibited MDM2 expression. The expression of MDM2 and P53 showed a negative correlation in NSCLC cell lines. MiR-221-3p down-regulation reduced the sensitivity of A549 cells to PTX, whereas its up-regulation partially reversed the A549/Taxol cells resistance to PTX and increased the chemosensitivity of A549/Taxol cells to PTX in xenograft models. Quantitative polymerase chain reaction analysis revealed that miR-221-3p expression increased, whereas the MDM2 level decreased in human NSCLC tumor tissues. Moreover, Western bolt analysis showed that P53 was lowly expressed in tumor tissues with MDM2 overexpression. Low expression of miR-221-3p in NSCLC tissues might indicate a poor T staging. In conclusion, miR-221-3p overexpression could regulate MDM2/p53 signaling pathway to reverse the PTX resistance of NSCLC and induce apoptosis in vitro and vivo.

Overexpression of hsa_circ_0002874 promotes resistance of non-small cell lung cancer to paclitaxel by modulating miR-1273f/MDM2/p53 pathway.

BACKGROUND: This study aimed to investigate the aberrant expression of hsa_circ_0002874 in non-small cell lung cancer (NSCLC) and elucidate associated molecular mechanisms that influence apoptosis and induce paclitaxel (PTX) resistance. METHODS: Inhibitors were used to downregulate circRNA or miRNA expression. pCDNA plasmid transfection and mimics were used to upregulate circRNA or miRNA expression. Dual-luciferase reporter assays were conducted to evaluate interactions between miR1273f and MDM2. Xenograft tumor models were used to assess the effect of hsa_circ_0002874 and miR1273f on tumor growth. NSCLC tissues and matched non-cancerous tissues were also collected for correlation analysis. RESULTS: hsa_circ_0002874 acts as a sponge for miR1273f which targets MDM2/P53. The stability of the hsa_circ_0002874/miR1273f/MDM2/P53 pathway was verified by upregulating and downregulating the expression of hsa_circ_0002874 and miR1273f. hsa_circ_0002874 downregulation or miR1273f upregulation reversed the resistance of the A549/Taxol cells in xenograft models. The expression of hsa_circ_0002874 was high, and the level of MDM2 was low in NSCLC tissues. P53 was only weakly expressed in NSCLC tissues with high expression of MDM2. CONCLUSIONS: hsa_circ_0002874 is strongly expressed in NSCLC tissues and maybe a potential marker for PTX resistance. hsa_circ_0002874 downregulation could regulate miR1273f/MDM2/P53 signaling pathway to reverse the PTX resistance of NSCLC and induce apoptosis in vitro and vivo.

A Microfluidic Sensor for Continuous, in Situ Surface Charge Measurement of Single Cells.

Cell surface charge has been recognized as an important cellular property. We developed a microfluidic sensor based on resistive pulse sensing to assess surface charge and sizes of single cells suspended in a continuous flow. The device consists of two consecutive resistive pulse sensors (RPSs) with identical dimensions. Opposite electric fields were applied on the two RPSs. A charged cell in the RPSs was accelerated or decelerated by the electric fields and thus exhibited different transit times passing through the two RPSs. The cell surface charge is measured with zeta potential that can be quantified with the transit time difference. The transit time of each cell can be accurately detected with the width of pulses generated by the RPS, while the cell size can be calculated with the pulse magnitude at the same time. This device has the ability to detect surface charges and sizes of individual cells with high tolerance in cell types and testing solutions compared with traditional electrophoretic light scattering methods. Three different types of cells including HeLa cancer cells, human dermal fibroblast cells, and human umbilical vein endothelial cells (HUVECs) were tested with the sensor. Results showed a significant difference of zeta potentials between HeLa cells and fibroblasts or HUVECs. In addition, when HeLa cells were treated with various concentrations of glutamine, the effects on cancer cell surface charge were detected. Our results demonstrated the great potential of using our sensor for cell type sorting, cancer cell detection, and cell status analysis.

Prognostic values of pretreatment neutrophil-to-lymphocyte and platelet-to-lymphocyte ratios in endometrial cancer: a systematic review and meta-analysis.

PURPOSE: Elevated inflammatory markers, including neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR), have been identified as poor predictors of survival in several malignancies. This meta-analysis was performed to quantify the effect of pretreatment NLR and PLR on the survival of patients with endometrial cancer (EC). METHODS: This review systematically searched for relevant publications in databases of PubMed, Embase, and the Cochrane Library. Pooled hazard ratios (pHRs) with 95% confidence intervals (95% CIs) were determined and used to explore the association between inflammatory markers and overall survival (OS) and disease-free survival (DFS) in a random-effects model. Subgroup analysis, sensitivity analysis, and publication bias were also conducted in this meta-analysis. RESULTS: Nine articles comprising 3390 patients were included. NLR higher than the cutoff was associated with a shorter OS (pHR = 2.22, 95% CI 1.77-2.78) and poorer PFS (pHR = 1.81, 95% CI 1.35-2.41). Patients with elevated PLR had high risk of decreased OS (pHR = 1.99, 95% CI = 1.51-2.61) and unfavorable PFS (pHR = 2.02, 95% CI 1.45-2.80). CONCLUSIONS: Elevated NLR and PLR during pretreatment are biomarkers of poor prognosis in patients with EC.

Enabling single cell electrical stimulation and response recording via a microfluidic platform.

Electrical stimulation (ES) has been recognized to play important roles in regulating cell behaviors. A microfluidic device was developed for the electrical stimulation of single cells and simultaneous recording of extracellular field potential (EFP). Each single cell was trapped onto an electrode surface by a constriction channel for ES testing and was then driven to the outlet by the pressure afterward. This design allows the application of ES on and detection of EFP of single cells continuously in a microfluidic channel. Human cardiomyocytes and primary rat cortex neurons were tested with specific ES with the device. Each cell's EFP signal was detected and analyzed during the ES process. Results have shown that after applying specific ES on the excitable single cells, the cells evoked electrical responses. In addition, increased secretion of glutamic acid was detected from the stimulated neurons. Altogether, these results indicated that the developed device can be used to continuously apply ES on and accurately determine cell responses of single cells with shorter probing time. The throughput of the measurement can achieve 1 cell per minute, which is higher than the traditional ES methods that need culturing cells or manually positioning the cells onto the electrode surface. Before and after the application of ES, the cell viability had no significant change. Such a device can be used to study the biological process of various types of cells under electrical stimulation.

Mortality of site-specific cancer in patients with schizophrenia: a systematic review and meta-analysis.

BACKGROUND: Numerous studies have reported contradicting results on the relationship between cancer mortality and schizophrenia. Our aim is to quantify the mortality rate of common site-specific cancers among patients with schizophrenia and to synthesize the available research evidence. METHODS: We performed a systemic search of the PubMed, EMBASE and Web of Science databases. Studies reporting the mortality rate of different cancer in patients with schizophrenia were included. A random-effects model was applied to calculate the pooled relative risks (RRs) with 95% confidence intervals (95%CIs). RESULTS: Seven studies consisting of 1,162,971 participants with schizophrenia were included in this meta-analysis. Data regarding mortality risk of breast, colon, lung and prostate cancer among schizophrenia patients were subjected to quantitative analysis. Pooled results showed significant increases in mortality risk of breast cancer (RR = 1.97, 95%CI 1.38-2.83), lung cancer (RR = 1.93, 95%CI 1.46-2.54) and colon cancer (RR = 1.69, 95%CI 1.60-1.80) in patients with schizophrenia compared with those in the general population or control group. The mortality risk of prostate cancer increased in male patients, although no significant difference was detected (RR = 1.58, 95% CI 0.79-3.15). Increased risks of mortality from lung and colon cancer were observed in female patients (RR = 2.49, 95%CI 2.40-2.59 and RR = 2.42, 95%CI 1.39-4.22, respectively) and elevated risks of mortality from lung and colon cancer in male patients (RR = 2.40, 95%CI 2.30-2.50 and RR = 1.90, 95%CI 1.71-2.11, respectively) were detected. CONCLUSIONS: Individuals with schizophrenia have a significantly high risk of mortality from breast, colon, and lung cancer.

Anticancer effect of berberine based on experimental animal models of various cancers: a systematic review and meta-analysis.

BACKGROUND: Numerous studies have explored the anti-tumor effect of berberine (BBR), but little clinical evidence guides the use of BBR in cancer patients. OBJECTIVES: Our aim was to investigate the impact of BBR on various cancers in healthy animals to promote the transformation from bench to bed. SEARCH METHODS: PubMed, Embase, Springer, and Cochrane databases were searched from January 2000 to October 2018 for relevant articles. SELECTION CRITERIA: Only published studies focusing on the relationship between BBR and various cancers in vivo were qualified. Two review authors independently assessed the risk of bias for each study, and any disagreement was resolved by discussion or by involving a third assessor. RESULTS: A total of 26 studies from 2000 to 2018, focusing on various cancer types, including breast cancer, liver cancer, colorectal cancer, nasopharyngeal carcinoma, lung cancer, gastric cancer, neuroepithelial cancer, endometrial carcinoma, esophageal cancer, tongue cancer, cholangiocarcinoma, and sarcoma were included. Overall, BBR reduced tumor volume (SMD =3.72, 95% CI: 2.89, 4.56, Z = 8.73, p < 0.00001) and tumor weight (SMD =2.35, 95% CI: 1.51, 3.19, Z = 5.50, p < 0.00001) in a linear The dose-response relationship (Pearson r = - 0.6717, p < 0.0001 in tumor volume analysis; Pearson r = - 0.7704, p < 0.0005 in tumor weight analysis). BBR inhibited angiogenesis in tumor tissues (SMD = 4.29, 95% CI: 2.14, 6.44, Z = 3.92, p < 0.00001), but it had no significant effect on the body weight of experimental animals (SMD = 0.11, 95% CI: - 0.70, 0.92, Z = 0.27, p = 0.78). Publication bias was not detected. CONCLUSION: BBR exerted anti-tumor effects in a variety of tumors in vivo, especially breast cancer and lung cancer, and the evidence was still insufficient in colorectal cancer and gastric cancer.

A microfluidic device for noninvasive cell electrical stimulation and extracellular field potential analysis.

We developed a device that can quickly apply versatile electrical stimulation (ES) signals to cells suspended in microfluidic channels and measure extracellular field potential simultaneously. The device could trap cells onto the surface of measurement electrodes for ES and push them to the downstream channel after ES by increasing pressure for continuous measurement. Cardiomyocytes, major functional cells in heart, together with human fibroblast cells and human umbilical vein endothelial cells, were tested with the device. Extracellular field potential signals generated from the cells were recorded. We found that under electrical stimulation, cardiomyocytes were triggered to alter their field potential, while non-excitable cells were not triggered. Hence this device can noninvasively distinguish electrically excitable cells from electrically non-excitable cells. Results have also shown that increased cardiomyocyte cell number led to increased magnitude and occurrence of the cell responses. This relationship could be used to detect the viable cells in a cardiac tissue. Application of variable ES signals on different cardiomyocyte clusters has shown that the application of ES clearly boosted cardiomyocytes electrical activities according to the stimulation frequency. In addition, we confirmed that the device can apply ES onto and detect the electrical responses from a mixed cell cluster; the responses from the mixed cluster is dependent on the ratio of cardiomyocytes. These results demonstrated that our device could be used as a tool to optimize ES conditions to facilitate the functional engineered cardiac tissue development.

Efficacy and safety of liposome-paclitaxel/liposome-paclitaxel combined with S-1 in 17 advanced gastric cancer patients with poor performance status.

BACKGROUND: To evaluate the efficacy and safety of liposome-paclitaxel (L-PTX)/L-PTX plus S-1 in advanced gastric cancer patients with poor performance status (PS). METHODS: We performed this retrospective study on 17 advanced gastric cancer patients with poor PS [rated as ≥2 based on the Eastern Cooperative Oncology Group (ECOG) scale] who underwent the following chemotherapy regimen: (I) L-PTX single-agent: L-PTX 60-80 mg/m2 given on days 1 and 8, in a 21-day cycle; (II) timed sequential (TS) regimen: L-PTX 60-80 mg/m2 given on days 1 and 8. S-1, 40-60 mg/m2 twice a day on days 1-14, in a 21-day cycle. Initially, some patients could not tolerate the 2-drug combination chemotherapy regimen, only L-PTX single-agent was given. After the patient's physical condition was improved, plus S-1 was also given. RESULTS: A total of 17 patients were studied. No complete response (CR) or partial response (PR) were observed in six patients, accounting for 35.29% (6/17). Stable disease (SD) was observed in five patients, accounting for 29.41% (5/17), and progressive disease (PD) in 6, accounting for 35.29% (6/17). The objective response and disease control rates were 35.29% (6/17) and 64.71% (11/17), respectively. The median progression-free survival (PFS) and median overall survival (OS) were 6.50 months [95% confidence interval (CI): 4.81-8.20] and 13.00 months (95% CI: 0.00-33.65), respectively. The most common hematological toxicities were neutropenia and anemia. CONCLUSIONS: L-PTX/L-PTX plus S-1 in the treatment of advanced gastric cancer patients with poor PS can prolong the patients' PFS and OS, and the toxicity is tolerable.

A microfluidic competitive immuno-aggregation assay for high sensitivity cell secretome detection.

We report a high-sensitivity cell secretome detection method using competitive immuno-aggregation and a micro-Coulter counter. A target cell secretome protein competes with anti-biotin-coated microparticles (MPs) to bind with a biotinylated antibody (Ab), causing decreased aggregation of the functionalized MPs and formation of a mixture of MPs and aggregates. In comparison, without the target cell secretome protein, more microparticles are functionalized, and more aggregates are formed. Thus, a decrease in the average volume of functionalized microparticles/aggregates indicates an increase in cell secretome concentration. This volume change is measured by the micro-Coulter counter, which is used to quantitatively estimate the cell secretome concentration. Vascular endothelial growth factor (VEGF), one of the key cell secretome proteins that regulate angiogenesis and vascular permeabilization, was used as the target protein to demonstrate the sensing principle. A standard calibration curve was generated by testing samples with various VEGF concentrations. A detection range from 0.01 ng/mL to 100.00 ng/mL was achieved. We further demonstrated the quantification of VEGF concentration in exogenous samples collected from the secretome of human mesenchymal stem cells (hMSCs) at different incubation times. The results from the assay agree well with the results of a parallel enzyme-linked immunoabsorbent assay (ELISA) test, indicating the specificity and reliability of the competitive immuno-aggregation assay. With its simple structure and easy sample preparation, this assay not only enables high sensitivity detection of VEGF but also can be readily extended to other types of cell secretome analysis as long as the specific Ab is known.

Lab-on-a-chip electrical multiplexing techniques for cellular and molecular biomarker detection.

Signal multiplexing is vital to develop lab-on-a-chip devices that can detect and quantify multiple cellular and molecular biomarkers with high throughput, short analysis time, and low cost. Electrical detection of biomarkers has been widely used in lab-on-a-chip devices because it requires less external equipment and simple signal processing and provides higher scalability. Various electrical multiplexing for lab-on-a-chip devices have been developed for comprehensive, high throughput, and rapid analysis of biomarkers. In this paper, we first briefly introduce the widely used electrochemical and electrical impedance sensing methods. Next, we focus on reviewing various electrical multiplexing techniques that had achieved certain successes on rapid cellular and molecular biomarker detection, including direct methods (spatial and time multiplexing), and emerging technologies (frequency, codes, particle-based multiplexing). Lastly, the future opportunities and challenges on electrical multiplexing techniques are also discussed.

Alanine and arginine rich domain containing protein, Aard, is directly regulated by androgen receptor in mouse Sertoli cells.

Alanine and arginine rich domain containing protein (Aard) is specifically expressed in Sertoli cells (SCs) of mouse testis and the expression increases in an age­dependent manner. A number of previous studies have indicated that androgen and androgen receptor (AR) signaling pathways are particularly important for spermatogenesis in mouse SCs, however, the association between Aard and AR remain to be elucidated. The present study identified Aard as a gene that is directly regulated by AR in mouse SCs, which is important in spermatogenesis. The expression of AARD was significantly downregulated in the testes of Sertoli cell­selective AR knockout mice compared with wild­type mice as analyzed by western blotting and immunofluorescence analyses. Quantitative polymerase chain reaction and western blotting indicated that AARD was predominantly expressed in adult mouse testis and its expression was increased in an age-dependent manner. In addition, AARD expression was upregulated by testosterone in primary SCs in vitro, which was confirmed by bioinformatics analysis and a dual­luciferase reporter assay. Finally, chromatin immunoprecipitation and electrophoretic mobility shift assays indicated that the ligand­bound AR activated Aard transcription via directly binding to the androgen­responsive element of the Aard promoter. To the best of our knowledge, the present study is the first to document that Aard is directly regulated by AR in mouse Sertoli cells.

A Novel Testis-Specific Gene, Ccdc136, Is Required for Acrosome Formation and Fertilization in Mice.

Testis-specific genes are essential for the spermatogenesis in mammalian male reproduction. In this study, we have identified a novel testis-specific gene, Ccdc136 (coiled-coil domain containing 136), from the results of high-throughput gene expression profiling in the developmental stage of mouse testes. Ccdc136 was conserved across species in evolution. Quantitative real-time polymerase chain reaction and Western blot analyses showed that Ccdc136 messenger RNA and protein were extraordinarily expressed in mouse testes, which was first presented at postnatal 3 week and increased in an age-dependent manner before adulthood. Immunofluorescence staining revealed that CCDC136 protein was most abundantly located in the acrosome of round spermatids and elongating spermatids within seminiferous tubules of the adult mouse testes. To investigate the function of Ccdc136 in mouse testes, we generated the Ccdc136-knockout mice using Cas9/RNA-mediated gene targeting technology. Interestingly, we found Ccdc136(-/-) males were infertile, due to severe defect of disrupting acrosome formation. The expression levels of proteins (SPACA1 and PICK1) involved in acrosome formation were significantly downregulated in the testes of Ccdc136(-/-) mice than wide-type mice. Moreover, in vitro fertilization assay revealed that anti-CCDC136 antibody could remarkably inhibit fertilization, suggesting CCDC136 also plays an important role in fertilization. All of these demonstrated the essential role of CCDC136-mediated acrosome formation in spermatogenesis and fertilization, which might also provide new insight into the genetic causes of human infertility.