Combinatorial optimization and spatial remodeling of CYPs to control product profile.

Activating inert substrates is a challenge in nature and synthetic chemistry, but essential for creating functionally active molecules. In this work, we used a combinatorial optimization approach to assemble cytochrome P450 monooxygenases (CYPs) and reductases (CPRs) to achieve a target product profile. By creating 110 CYP-CPR pairs and iteratively screening different pairing libraries, we demonstrated a framework for establishing a CYP network that catalyzes six oxidation reactions at three different positions of a chemical scaffold. Target product titer was improved by remodeling endoplasmic reticulum (ER) size and spatially controlling the CYPs' configuration on the ER. Out of 47 potential products that could be synthesized, 86% of the products synthesized by the optimized network was our target compound quillaic acid (QA), the aglycone backbone of many pharmaceutically important saponins, and fermentation achieved QA titer 2.23 g/L.

Changes of arachnoid granulations after subarachnoid hemorrhage in cynomolgus monkeys.

This research explores ultrastructural changes of arachnoid granulations associated with hydrocephalus after subarachnoid hemorrhage in cynomolgus monkeys. It provides a theoretical basis for further study of the etiology and prevention of hydrocephalus. Female cynomolgus monkeys about one-year-old were selected. The position range of arachnoid granulations in superior sagittal sinus and transverse sinus was determined in a randomly selected control monkey. The morphology of normal arachnoid granulations in cynomolgus monkeys was observed under a transmission electron microscope. A primate model of subarachnoid hemorrhage was established by injecting autologous blood into cisterna magna. Vomiting, movement disorder, and reduced level of consciousness were gradually observed in monkeys. Computed tomography and magnetic resonance imaging scan results confirmed subarachnoid hemorrhage and hydrocephalus, and the morphology of arachnoid granulations in hydrocephalus was observed under a transmission electron microscope. Extensive fibrosis of arachnoid granulations was observed under a transmission electron microscope in cynomolgus monkeys with hydrocephalus after subarachnoid hemorrhage.

Bacterial Single Cell Whole Transcriptome Amplification in Microfluidic Platform Shows Putative Gene Expression Heterogeneity.

Single cell RNA sequencing is a technology that provides the capability of analyzing the transcriptome of a single cell from a population. So far, single cell RNA sequencing has been focused mostly on human cells due to the larger starting amount of RNA template for subsequent amplification. One of the major challenges of applying single cell RNA sequencing to microbial cells is to amplify the femtograms of the RNA template to obtain sufficient material for downstream sequencing with minimal contamination. To achieve this goal, efforts have been focused on multiround RNA amplification, but would introduce additional contamination and bias. In this work, we for the first time coupled a microfluidic platform with multiple displacement amplification technology to perform single cell whole transcriptome amplification and sequencing of Porphyromonas somerae, a microbe of interest in endometrial cancer, as a proof-of-concept demonstration of using single cell RNA sequencing tool to unveil gene expression heterogeneity in single microbial cells. Our results show that the bacterial single-cell gene expression regulation is distinct across different cells, supporting widespread heterogeneity.

Effects of over-expression of SOD2 in bone marrow-derived mesenchymal stem cells on traumatic brain injury.

Intravenous administration of bone marrow-derived mesenchymal stem cells (BM-MSCs) has been shown to promote nerve cell regeneration following traumatic brain injury (TBI). As the anti-oxidant defense systems in neuronal tissue including superoxide dismutase 2 (SOD2) are crucial to defend cell against oxidative stress. We proposed a new stratege to increase the therapeutic effect of MSCs by preventing cells death from oxidative stress. We overexpressed SOD2 in BM-MSCs, transplanted these MSCs into TBI model mice, assessed the protective effect of SOD2 against oxidation-induced apoptosis in BM-MSCs both in vitro and in vivo, evaluated brain functional recovery by the rotarod behavioral test, and tested the oxidation status of TBI mice brain after BM-MSCs transplantation by monitoring the superoxide dismutase, glutathione and malonaldehyde level. We found over-expression of SOD2 protected BM-MSCs from H2O2-induced cell apoptosis. Injection of SOD2 over-expressed BM-MSCs attenuated neuro-inflammation in the ipsilateral cortex of TBI mice, and protected TBI mice against loss of blood-brain barrier integrity. Furthermore, the rotarod behavioral test showed functional recovery of TBI mice after MSC treatment. Our experiments indicated that SOD2-over-expressed BM-MSCs have an improved therapeutic effect on brain injury treatment in TBI mice.

Role of Peroxiredoxin 2 in the Protection Against Ferrous Sulfate-Induced Oxidative and Inflammatory Injury in PC12 Cells.

Peroxiredoxin 2 (Prdx2) is a ubiquitous antioxidant enzyme in mammalian brain. Although a protective role of Prdx2 has been established in cerebral ischemia and several neurodegenerative diseases, its contribution against iron-induced neurocytotoxicity still remains to be determined. Accordingly, in this study, we aimed to investigate the effects of Prdx2 on iron-induced cytotoxicity using an in vitro model in which PC12 cells are exposed to ferrous sulfate (FS). The FS treatment increased Prdx2 expression, and promoted lactate dehydrogenase (LDH) release and cell apoptosis in PC12 cells, accompanied by the increase in the Bax/Bcl2 ratio, cytochrome c release, and caspase-3 cleavage. FS exposure also increased the malondialdehyde content (lipid peroxidation), 3'-nitrotyrosine expression (protein nitration), γ-H2A.X formation (DNA oxidation), and promoted nuclear factor kappa B nuclear translocation, cyclooxygenase-2 expression, and release of tumor necrosis factor-α and interleukin-1ß. Lentivirus-mediated Prdx2 knockdown intensified the FS-induced LDH release and cell apoptosis by aggravating the oxidative and inflammatory damage. In conclusion, our findings demonstrated that Prdx2 played a vital role in the protection against iron-induced cytotoxicity in PC12 cells.

CacyBP/SIP inhibits the migration and invasion behaviors of glioblastoma cells through activating Siah1 mediated ubiquitination and degradation of cytoplasmic p27.

Calcyclin-binding protein or Siah-1-interacting protein (CacyBP/SIP) has been reported to be up-regulated and plays an important role in promoting cell proliferation in human glioma. However, the effect of CacyBP/SIP on glioma cell motility is still unclear. Here, to our surprise, CacyBP/SIP was found to inhibit the migration and invasion of glioma cells U251 and U87. Silencing of CacyBP/SIP significantly promoted the migration and invasion behaviors of glioma cells. On the contrary, overexpression of CacyBP/SIP obviously suppressed them. Further investigation indicated that silencing of CacyBP/SIP significantly reduced the interaction between Siah1 and cytoplasmic p27, which in turn attenuated the ubiquitination and degradation of cytoplasmic p27. In contrast, overexpression of CacyBP/SIP promoted the interaction between Siah1 and cytoplasmic p27, which in turn increased the ubiquitination and degradation of cytoplasmic p27. Importantly, the degradation of p27 could be blocked by Siah1 knockdown. Finally, we found that CacyBP/SIP was reversely related to cytoplasmic p27 in human normal brain tissues and glioma tissues. Taken together, these results suggest that CacyBP/SIP plays an important role in inhibiting glioma cell migration and invasion through promoting the degradation of cytoplasmic p27.

Increased Expression of T Cell Immunoglobulin and Mucin Domain 3 on CD14+ Monocytes Is Associated with Systemic Inflammatory Reaction and Brain Injury in Patients with Spontaneous Intracerebral Hemorrhage.

OBJECTIVE: To study the expression of T cell immunoglobulin and mucin domain 3 (Tim-3) on peripheral blood immunocytes, and the relationship between Tim-3 and the systemic inflammatory response or brain injury in patients with intracerebral hemorrhage (ICH). METHODS: According to the volume of hematoma at 12 hours after onset of ICH, 60 newly diagnosed patients with ICH were divided into the small (volume of hematoma <30 mL, 30 cases) and large (volume of hematoma ≥30 mL, 30 cases) ICH groups. The expression of Tim-3 on peripheral blood immunocytes was analyzed by flow cytometry. Real-time reverse transcriptase polymerase chain reaction was used to detect Tim-3 mRNA on peripheral blood mononuclear cells (PBMCs). Meanwhile, tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), and S-100B protein were measured by enzyme-linked immunosorbent assay. Glasgow outcome scale (GOS) score at Day 30 was used to estimate prognosis of patients. RESULTS: The leukocyte count, neutrophil count, monocyte count, TNF-α, IL-1ß, and S-100B protein increased remarkably after ICH. However, all of them in the large ICH group increased more obviously, and there were significant differences when compared with those in the small ICH group (P < .01). The expression of Tim-3 mRNA on PBMCs in the large ICH group increased remarkably, peaked at Day 3, and was positively associated with the concentrations of TNF-α, IL-1ß, and S-100B protein (P < .01). Tim-3 was predominantly expressed itself on CD14+ monocytes. There was a negative correlation between GOS score and Tim-3 mRNA, TNF-α, IL-1ß, or S-100B protein. CONCLUSIONS: The expression of Tim-3 on CD14 + monocytes involves in systemic inflammatory reaction after ICH and may be a novel treatment target.

RBM5 inhibits tumorigenesis of gliomas through inhibition of Wnt/ß-catenin signaling and induction of apoptosis.

BACKGROUND: Gliomas are one of the most common malignant brain tumors and bring a big threat to human life as traditional therapy is unsatisfactory. RBM5 was a RNA-binding motif protein and was reported as a tumor suppressor. But the role of RBM5 in gliomas was unknown. METHODS: The mRNA level of RBM5 was determined in gliomas tissues and cell lines by real-time quantitative PCR (qRT-PCR) assay while the association of RBM5 expression with prognosis was analyzed by Kaplan-Meier method and compared by log-rank test. Lentivirus was used to overexpress RBM5 in gliomas cells. MTT and BrdU incorporation assay were used to determine cell proliferation and DNA synthesis when the ability of cell migration and invasion was analyzed by transwell assay with/without Matrigel. Cell apoptosis rate was determined with fluorescence-activated cell sorting (FACS) method. Then, expression of apoptosis molecules and critical members in Wnt/ß-catenin pathway were detected by western blot analysis. RESULTS: RBM5 was shown to be downregulated in gliomas tissues and gliomas cell lines. And decreased RBM5 expression was clinically correlated with tumor stage, patient age, and poor prognosis of gliomas patients. The proliferation and DNA synthesis was dramatically inhibited when RBM5 was overexpressed in SHG44 or U251 cells. Also, the ability of cell migration and invasion was disrupted. Then, the level of ß-catenin and Cyclin D1 significantly decreased when DKK1 and P-GSK-3ß increased reversely in SHG44 cells, which suggested that RBM5 inhibited canonical Wnt/ß-catenin signaling. Meanwhile, we demonstrated that caspase3-mediated apoptotic pathway was activated by RBM5 as Bax, TNF-α, and cleaved caspase3 were greatly upregulated while antiapoptotic molecule Bcl-2 was downregulated. Additionally, that apoptotic rate increased significantly from less than 1 to 32% in RBM5-overexpressed SHG44 cells further supported the pro-apoptosis role of RBM5 in gliomas cells. CONCLUSIONS: RBM5 plays a suppressor role in human gliomas by inhibiting Wnt/ß-catenin signaling and inducing cell apoptosis. This study improves our knowledge about the carcinogenesis and progression of human gliomas, which would greatly contribute to the therapy for gliomas patients.

Nuclear factor-κB activation in perihematomal brain tissue correlates with outcome in patients with intracerebral hemorrhage.

BACKGROUND: Nuclear factor-κB (NF-κB) plays an important role in the inflammatory response after intracerebral hemorrhage (ICH). We therefore proposed that NF-κB activation in perihematomal brain tissue might correlate with clinical outcome in patients with ICH. To confirm this, we studied clinical data of 45 patients with ICH and NF-κB activation in perihematomal brain tissue and analyzed predictors of clinical outcome as well as the predictive value of NF-κB activation. METHODS: Forty-five patients with spontaneous basal ganglia hemorrhage were prospectively investigated. The clinical data were collected, which include demographics, alcohol and tobacco abuse, stroke risk factors, neuroimaging variables at presentation, Glasgow Coma Scale (GCS) at admission, number of days in hospital, mechanical ventilation, pneumonia, and outcome. Clinical outcome was assessed by the modified Rankin Scale at 6 months after ICH. Perihematomal brain tissue was collected, and NF-κB activation was detected using immunohistochemistry. Univariate analysis and multivariate logistic regression analysis were performed to determine predictors of the poor outcome. RESULTS: Immunohistochemical detection showed that NF-κB p65 was expressed in the nuclei of neurons and glial cells in all patients. The number of nuclear NF-κB p65-positive cells was 54 ± 21. Six months after ICH, 18 (40%) patients achieved a favorable functional outcome (mRS ≤ 3) while 27 (60%) had a poor functional outcome (mRS 4 to 6). In univariate analysis, predictors of poor functional outcome were lower GCS score on admission (P = 0.004), larger hematoma volume (P = 0.004), intraventricular extension (P = 0.047), midline shift (P = 0.005), NF-κB activation (P < 0.0001), mechanical ventilation (P = 0.018), and co-morbidity with pneumonia (P = 0.002). In multivariate logistic regression analysis, NF-κB activation was the only independent predictor of poor outcome at 6 months after ICH. CONCLUSIONS: NF-κB activation is closely related to clinical outcome 6 months after ICH in humans. Therefore, it could be useful to predict prognosis of ICH accurately and should be further evaluated as a target for therapeutic strategies of ICH in the future.

Chronic subdural haematoma evolving from traumatic subdural hydroma.

OBJECTIVE: This study aimed to investigate the incidence and clinical characteristics of chronic subdural haematoma (CSDH) evolving from traumatic subdual hydroma (TSH). METHODS: The clinical characteristics of 44 patients with CSDH evolving from TSH were analysed retrospectively and the relevant literature was reviewed. RESULTS: In 22.6% of patients, TSH evolved into CSDH. The time required for this evolution was 14-100 days after injury. All patients were cured with haematoma drainage. CONCLUSIONS: TSH is one possible origin of CSDH. The clinical characteristics of TSH evolving into CSDH include polarization of patient age and chronic small effusion. The injuries usually occur during deceleration and are accompanied by mild cerebral damage.

MiR-7-5p is frequently downregulated in glioblastoma microvasculature and inhibits vascular endothelial cell proliferation by targeting RAF1.

The aberrant expression of microRNAs (miRNAs) is always associated with tumor development and progression. Microvascular proliferation is one of the unique pathologic features of glioblastoma (GBM) . In this study, the microvasculature from GBM or normal brain tissue derived from neurosurgeries was purified and total RNA was isolated from purified microvasculature. The difference of miRNA expression profiles between glioblastoma microvasculature and normal brain capillaries was investigated. It was found that miR-7-5p in GBM microvessels was significantly reduced compared with that in normal brain capillaries. In the in vitro experiments, overexpression of miR-7-5p significantly inhibited human umbilical vein endothelial cell proliferation. Forced expression of miR-7-5p in human umbilical vein endothelial cells in vitro significantly reduced the protein level of RAF1 and repressed the activity of the luciferase, a reporter vector carrying the 3'-untranslated region of RAF1. These findings indicate that RAF1 is one of the miR-7-5p target genes. Furthermore, a significant inverse correlation between miR-7-5p expression and RAF1 protein level in GBM microvasculature was found. These data suggest that miR-7-5p functions as a tumor suppressor gene to regulate GBM microvascular endothelial cell proliferation potentially by targeting the RAF1 oncogene, implicating an important role for miR-7-5p in the pathogenesis of GBM. It may serve as a guide for the antitumor angiogenesis drug development.

NF-κB activation and cell death after intracerebral hemorrhage in patients.

Nuclear factor-κB (NF-κB) plays an important role in secondary damage after intracerebral hemorrhage (ICH). We explored NF-κB activation and the relationship between NF-κB and cell death in the perihematomal brain tissue of patients after ICH. According to the interval between onset of hemorrhage and specimen collection, 53 cases of patients with basal ganglia hemorrhage were divided into six experimental groups: 0-6, 7-12, 13-24, 25-48, 49-96, and >96 h group. Brain tissues of the experimental groups and control group were collected. IL-1ß, TNF-α, and NF-κB p65 expressions at the protein level were detected by immunohistochemistry. Cell death was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay. All of the detection items of immunohistochemistry and TUNEL showed significant differences between the experimental groups and control group. At the protein level, nuclear NF-κB p65, IL-1ß, and TNF-α achieved maximum values at 13-48, 0-24, and 13-48 h, respectively. Maximum cell death was reached at 13-48 h. NF-κB activation increased dramatically in perihematomal brain tissue after ICH. NF-κB activation was closely related with cell death and had an important function in secondary brain damage after ICH in patients.

[Clinical characteristics of primary extradural meningiomas in head].

OBJECTIVE: To explore the clinical characteristics, efficacy and prognosis of primary extradural meningiomas (PEMs) in head. METHODS: A total of 19 treated surgically case of PEMs at Qilu Hospital, Shandong University between May 1994 and December 2012 were analyzed retrospectively with respects to their demographic features, presenting symptoms & duration, tumor location & type, imaging features, surgical results, pathological grade, histological subtype and follow-up outcomes. RESULTS: PEMs in head accounted for 1.2% of all meningiomas in this group. The male-to-female ratio was 1: 0.7. The mean age was 36.6 years with a bimodal distribution of age. The common presenting symptoms included nasal obstruction and a painless and gradually expanding mass in the region of lesion. The average duration of symptom was 3.07 years. The skull convexities, paranasal sinuses and nasal cavity, orbit and epidural space were common tumor sites. The most common type was typeII. The rate of total tumor removal was 100%. And no perioperative mortality occurred in this series. Benign and atypical meningiomas accounted for 94.7% and 6.3% respectively. Meningothelial and psammomatous meningiomas were common histopathological subtypes. There was one case of tumoral recurrence. And no mortality was reported during a mean follow-up period of 2.43 (0.25-8.5) years. CONCLUSION: PEMs in head have some marked clinical characteristics compared with primary intradural meningiomas. Total tumor removal together with a wide excision of all involved tissue followed by the reconstruction of tissue defects is the best surgical option. The prognosis is excellent in most cases after complete resection.

A digital microfluidic device integrated with electrochemical sensor and 3D matrix for detecting soluble PD-L1.

PD1/PD-L1 checkpoint inhibitors are at the forefront of cancer immunotherapies. However, the overall response rate remains only 10-30%. Even among initial responders, drug resistance often occurs, which can lead to prolonged use of a futile therapy in the race with the fatal disease. It would be ideal to closely monitor key indicators of patients' immune responsiveness, such as circulating PD-L1 levels. Traditional PD-L1 detection methods, such as ELISA, are limited in sensitivity and rely on core lab facilities, preventing their use for the regular monitoring. Electrochemical sensors exist as an attractive candidate for point-of-care tool, yet, streamlining multiple processes in a single platform remains a challenge. To overcome this challenge, this work integrated electrochemical sensor arrays into a digital microfluidic device to combine their distinct merits, so that soluble PD-L1 (sPD-L1) molecules can be rapidly detected in a programmed and automated manner. This new platform featured microscale electrochemical sensor arrays modified with electrically conductive 3D matrix, and can detect as low as 1 pg/mL sPD-L1 with high specificity. The sensors also have desired repeatability and can obtain reproducible results on different days. To demonstrate the functionality of the device to process more complex biofluids, we used the device to detect sPD-L1 molecules secreted by human breast cancer cell line in culture media directly and observed 2X increase in signal compared with control experiment. This novel platform holds promise for the close monitoring of sPD-L1 level in human physiological fluids to evaluate the efficacy of PD-1/PD-L1 immunotherapy.

Increased expression of T cell immunoglobulin and mucin domain 3 aggravates brain inflammation via regulation of the function of microglia/macrophages after intracerebral hemorrhage in mice.

BACKGROUND: Microglia/macrophages are known to play important roles in initiating brain inflammation after spontaneous intracerebral hemorrhage (ICH). T cell immunoglobulin and mucin domain-3 (Tim-3) have been proven to play a critical part in several inflammatory diseases through regulation of both adaptive and innate immune responses. Tim-3 can be expressed by microglia/macrophages and regulates their function in the innate immune response. However, the effect of Tim-3 on inflammatory responses following ICH is unclear. METHODS: In this study, we investigated Tim-3 expression, the inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß), and brain water content in peri-hematomal brain tissue at 12 hours and at 1, 3, 5, and 7 days post-ICH in wild type (WT) ICH and Tim-3-/- ICH mice. The numbers of Tim-3 positive cells,astrocytes, neutrophils and microglia/macrophages were detected using immunofluorescence staining. Cytokines were measured by ELISA. Double immunofluorescence labeling was performed to identify the cellular source of Tim-3 expression. Mouse neurological deficit scores were assessed through animal behavior. RESULTS: Expression of Tim-3 increased early in mouse peri-hematomal brain tissue after autologous blood injection, peaked at day 1, and was positively correlated with the concentrations of TNF-α, IL-1ß, and brain water content. Tim-3 was predominantly expressed in microglia/macrophages. Compared with WT mice, Tim-3-/- mice had reduced ICH-induced brain inflammation with decreased TNF-α and IL-1ß, cerebral edema and neurological deficit scores. Moreover, Tim-/- inhibited activation of microglia/macrophages. The number of activated microglia/macrophages in Tim-3-/- ICH mice was much lower than that in WT ICH mice. CONCLUSIONS: Our findings demonstrate that Tim-3 plays an important role in brain inflammation after ICH, and may be a potential treatment target.

Precise antibacterial therapeutics based on stimuli-responsive nanomaterials.

Bacterial infection refers to the process in which bacteria invade, grow, reproduce, and interact with the body, ultimately causing a series of pathological changes. Nowadays, bacterial infection remains a significant public health issue, posing a huge threat to human health and a serious financial burden. In the post-antibiotic era, traditional antibiotics are prone to inducing bacterial resistance and difficulty in removing bacterial biofilm. In recent years, antibacterial therapy based on nanomaterials has developed rapidly. Compared with traditional antibiotics, nanomaterials effectively remove bacterial biofilms and rarely result in bacterial resistance. However, due to nanomaterials' strong permeability and effectiveness, they will easily cause cytotoxicity when they are not controlled. In addition, the antibacterial effect of non-responsive nanomaterials cannot be perfectly exerted since the drug release property or other antibacterial effects of these nano-materials are not be positively correlated with the intensity of bacterial infection. Stimuli-responsive antibacterial nanomaterials are a more advanced and intelligent class of nano drugs, which are controlled by exogenous stimuli and microenvironmental stimuli to change the dosage and intensity of treatment. The excellent spatiotemporal controllability enables stimuli-responsive nanomaterials to treat bacterial infections precisely. In this review, we first elaborate on the design principles of various stimuli-responsive antibacterial nanomaterials. Then, we analyze and summarizes the antibacterial properties, advantages and shortcomings of different applied anti-bacterial strategies based on stimuli-responsive nanomaterials. Finally, we propose the challenges of employing stimuli-responsive nanomaterials and corresponding potential solutions.

Protocol for photoelectrocatalytic synthesis of aromatic azo compounds from aromatic amines.

Photoelectrocatalytic (PEC) strategy has emerged as a promising approach to drive organic reactions under mild conditions. Here, we present a protocol for PEC oxidative coupling of aromatic amines to produce aromatic azo compounds over a porous BiVO4 nanoarray (BiVO4-NA) photoanode. We describe the fabrication of BiVO4-NA photoanode and the detailed steps for the PEC oxidative coupling reaction, including key performance data of the BiVO4-NA photoanode for synthesizing azobenzene from aniline. For complete details on the use and execution of this protocol, please refer to Luo et al. (2022).1.

Current Status and Emerging Trends in Colorectal Cancer Screening and Diagnostics.

Colorectal cancer (CRC) is a prevalent and potentially fatal disease categorized based on its high incidences and mortality rates, which raised the need for effective diagnostic strategies for the early detection and management of CRC. While there are several conventional cancer diagnostics available, they have certain limitations that hinder their effectiveness. Significant research efforts are currently being dedicated to elucidating novel methodologies that aim at comprehending the intricate molecular mechanism that underlies CRC. Recently, microfluidic diagnostics have emerged as a pivotal solution, offering non-invasive approaches to real-time monitoring of disease progression and treatment response. Microfluidic devices enable the integration of multiple sample preparation steps into a single platform, which speeds up processing and improves sensitivity. Such advancements in diagnostic technologies hold immense promise for revolutionizing the field of CRC diagnosis and enabling efficient detection and monitoring strategies. This article elucidates several of the latest developments in microfluidic technology for CRC diagnostics. In addition to the advancements in microfluidic technology for CRC diagnostics, the integration of artificial intelligence (AI) holds great promise for further enhancing diagnostic capabilities. Advancements in microfluidic systems and AI-driven approaches can revolutionize colorectal cancer diagnostics, offering accurate, efficient, and personalized strategies to improve patient outcomes and transform cancer management.

[Expression and significance of MMP-9, PR, Ki-67 and Survivin in multiple meningiomas].

OBJECTIVE: To detect the expressions of matrix metalloproteinase-9(MMP-9), progesterone receptor(PR), Ki-67 and Survivin in the multiple meningiomas (MMs) and explore its genesis, diagnosis and bionomics features. METHODS: A total of 66 cases with histological sections of meningiomas retrieved from the archives of Department of Pathology of our hospital, including 30 cases of solitary meningiomas (SMs) and 36 cases MMs, were regrouped as benign, atypical and malignant by hematoxylin and eosin staining according to the World Health Organization classification of nervous system tumors. Immunohistochemistry was performed to detect the expressions of MMP-9, PR, Ki-67 and Survivin in 36 cases of MMs and 30 cases of SMs. And normal brain tissue was selected as a control group. The staining intensity was analyzed quantitatively for the differential expressions of MMP-9, PR, Ki-67 and Survivin between SMs and MMs. RESULTS: No expression of MMP-9, PR, Ki-67 and Survivin was detected in 5 normal brain tissues, but the expression rates were 100%, 53%, 23% and 88% respectively for significant difference comparing with normal tissue. The result of statistical analysis showed that there was significant difference in the expression intensity of MMP-9 and PR between two groups. The expression intensity MMP-9 in multiple group was significantly higher in MMs than that in SMs (P < 0.01) while PR was lower in MMs than that in SMs (P < 0.05). But no significant difference was found for the expression of Ki-67 or Survivin between two groups. CONCLUSION: The detections of MMP-9, PR, Ki-67 and Survivin are helpful in the clinical diagnosis and early detection of meningioma.

[Aberrant methylation of NF2, TIMP-3 and THBS1 genes and their diagnostic values in meningiomas].

OBJECTIVE: To explore the functions of NF2, TIMP-3 and THBS1 genes in the tumorigenesis or progression of meningiomas and analyze the values of these genes in early diagnosis, therapy and prognostic evaluation in meningiomas. METHODS: A total of 66 cases with histological sections of meningiomas, including solitary (SMs, n = 30) and multiple meningiomas (MMs, n = 36), were retrieved from our departmental archives. All cases were regrouped as benign, atypical and anaplastic (malignant) by hematoxylin & eosin staining according to the recently published WHO classification of nervous system tumors. Genomic DNA was extracted from tumor sections and methylation-specific polymerase chain reaction (MSP) performed to detect the CpG methylation status. Normal brain tissue was used as the control group. And then the differences of methylation rate between SMs and MMs tissues and among different subgroups were analyzed by statistical analyses. RESULTS: The results of methylation in different types of meningiomas demonstrated that the rates of NF2, TIMP-3 and THBS1 methylation were 26.7% (8/30), 16.7% (5/30) and 36.7% (11/30) in 30 SMs tissues and 30.6% (11/36), 22.2% (8/36) and 22.2% (8/36) in 36 MMs tissues respectively. But no aberrant methylation of NF2, TIMP-3 and THBS1 genes was found in normal brain tissue. No significant differences in three types of gene methylation rates existed between SMs and MMs in the I-III grade meningiomas. Nevertheless, there was great difference between grades I, II and III in SMs and MMs while no significant difference was found between grades II and III. CONCLUSION: The methylation of NF2, TIMP-3 and THBS1 is correlated with the tumorigenesis of meningiomas (grade II and III). As an important pathogenetic cause of meningiomas, it may be used as a clinical tool for an early diagnosis of meningiomas.