A highly sensitive Golgi-targeted fluorescent probe for the simultaneous detection of malondialdehyde and formaldehyde in living systems and foods.

Malondialdehyde (MDA) and formaldehyde (FA) are highly active carbonyl substances widely present in both biological and abiotic systems. The detection of MDA and FA is of great significance for disease diagnosis and food safety monitoring. However, due to the similarity in structural properties between MDA and FA, very few probes for synergistically detecting MDA and FA were reported. In addition, functional abnormalities in the Golgi apparatus are closely related to MDA and FA, but currently there are no fluorescent probes that can detect MDA and FA in the Golgi apparatus. Therefore, we constructed a simple Golgi-targetable fluorescent probe GHA based on hydrazine moiety as the recognition site to produce a pyrazole structure after reaction with MDA and to generate a CN double bond after reaction with FA, allowing MDA and FA to be distinguished due to different emission wavelengths during the recognition process. The probe GHA has good specificity and sensitivity. Under the excitation of 350 nm, the blue fluorescence was significantly enhanced at 424 nm when the probe reacted with MDA, and the detection limit was 71 nM. At the same time, under the same excitation of 350 nm, the reaction with FA showed a significant enhancement of green fluorescence at 520 nm, with a detection limit of 12 nM for FA. And the simultaneous and high-resolution imaging of MDA and FA in the Golgi apparatus of cells was achieved. In addition, the applications of the probe GHA in food demonstrated it can provide a powerful method for food safety monitoring. In summary, this study offers a promising tool for the synergistic identification and determination of MDA and FA in the biosystem and food, facilitating the revelation of their detailed functions in Golgi apparatus and the monitoring of food safety.

Dissecting lysosomal viscosity fluctuations in live cells and liver tissues with an ingenious NIR fluorescent probe.

Viscosity is a pivotal component in the cell microenvironment, while lysosomal viscosity fluctuation is associated with various human diseases, such as tumors and liver diseases. Herein, a near-infrared fluorescent probe (BIMM) based on merocyanine dyes was designed and synthesized for detecting lysosomal viscosity in live cells and liver tissue. The increase in viscosity restricts the free rotation of single bonds, leading to enhanced fluorescence intensity. BIMM exhibits high sensitivity and good selectivity, and is applicable to a wide pH range. BIMM has near-infrared emission, and the fluorescent intensity shows an excellent linear relationship with viscosity. Furthermore, BIMM possessing excellent lysosomes-targeting ability, and can monitor viscosity changes in live cells stimulated by dexamethasone, lipopolysaccharide (LPS), and nigericin, and differentiate between cancer cells and normal cells. Noticeably, BIMM can accurately analyze viscosity changes in various liver disease models with HepG2 cells, and is successfully utilized to visualize variations in viscosity on APAP-induced liver injury. All the results demonstrated that BIMM is a powerful wash-free tool to monitor the viscosity fluctuations in living systems.

A ratiometric fluorescent probe for imaging the fluctuation of HOBr during endoplasmic reticulum stress.

Endoplasmic reticulum (ER) stress is closely associated with cell apoptosis, autophagy, DNA damage, metabolism, and migration. When ER stress occurs, a large number of reactive oxygen species, including hypobromous acid (HOBr), are generated. The degree of ER stress can be understood by accurately detecting the HOBr concentration in the ER. Unfortunately, no ER-targetable probes for detecting HOBr have been reported to date. To solve this problem, we developed a naphthalimide-based fluorescent probe (ER-NABr) for imaging HOBr in the ER. Upon reaction with HOBr, a red shift in the fluorescence spectrum occurs due to the difference in the molecular conjugation between the original ER-NABr and the reaction product. ER-NABr showed a fast response (within 30 s) and high selectivity towards HOBr, with a ratiometric quantitative response (5-40 µM) and high sensitivity (138 nM). With its excellent biocompatibility and remarkable ER-targetable ability, ER-NABr was successfully utilized to ratiometrically image intracellular HOBr, particularly during ER stress, which is beneficial for revealing the role of HOBr in ER-associated diseases.

A turn-on fluorescent probe for detecting and bioimaging of HOCl in inflammatory and liver disease models.

Hypochlorous acid (HOCl) is a common reactive oxygen species (ROS) associated with the development of liver, tumor, inflammatory, and other diseases. In this work, the turn-on fluorescent probe named (WZ-HOCl) with a naphthalimide structure was designed and synthesized to detect endogenous HOCl in disease models. WZ-HOCl can achieve a fast response to HOCl with good linearity in the range of 0-45 µM (LOD = 147 nM). The application of WZ-HOCl in bioimaging was investigated by constructing a series of cellular disease models, and the results showed that WZ-HOCl could sensitively detect endogenous HOCl in inflammatory and liver disease models. It can also be used to differentiate between hepatocytes and hepatoma cells. WZ-HOCl will provide new methods and ideas for fluorescent probes in detecting drug-induced liver injury, alcoholic and non-alcoholic steatohepatitis, and some inflammation-related diseases.

A thiomorpholine substituted malonyl-coumarin dye for discriminative detection of hydrazine and strong acidity.

Detection of toxic hydrazine and harmful strong acidity is of great importance for survival of organisms. In the present paper, a new thiomorpholine substituted malonyl-coumarin dye was synthesized for discriminative detection of hydrazine and strong acidity. At pH 7.4, the fluorescence at 560 nm decreased and that at 496 nm increased upon reaction with hydrazine, which was used for on-site detection of hydrazine vapor and endogenous hydrazine in live cells. From pH 2.0 to 1.2, the fluorescence at 563 nm increased greatly, which could be ascribed to the PET process from thiomorpholine to malonyl-coumarin. The probe was desirable for discriminative detection of toxic hydrazine and strong acidity.

Tibial plateau fracture and RNA sequencing with osteogenesis imperfecta: a case report.

Osteogenesis imperfecta (OI) is a hereditary skeletal dysplasia with an incidence of approximately 1:15,000 to 20,000. OI is usually caused by the mutation of COL1A1 and COL1A2, which would encode the α-chain of type I collagen. OI is clinically characterized by decreased bone mass, increased risk of bone fragility, blue sclerae, and dentinogenesis. Case presentation: A 29-year-old male patient was diagnosed with right tibial plateau fracture caused by slight violence. Physical examination revealed the following: height, 140 cm; weight, 70 kg; body mass index (BMI), 35.71 kg/m2; blue sclera and barrel chest were observed. X-ray examination showed left convex deformity of the thoracic vertebrae with reduced thoracic volume. Laboratory examinations revealed a decrease in both vitamin D and blood calcium levels. Bone mineral density (BMD) was lower than the normal range. After the preoperative preparation was completed, the open reduction and internal fixation of the right tibial plateau fracture were performed. Meanwhile, whole blood samples of this OI patient and the normal control were collected for RNA transcriptome sequencing. The RNA sequence analysis revealed that there were 513 differentially expressed genes (DEGs) between this OI patient and the normal control. KEGG-enriched signaling pathways were significantly enriched in extracellular matrix (ECM)-receptor interactions. Conclusion: In this case, DEGs between this OI patient and the normal control were identified by RNA transcriptome sequencing. Moreover, the possible pathogenesis of OI was also explored, which may provide new evidence for the treatment of OI.

Evolved Biosensor with High Sensitivity and Specificity for Measuring Cadmium in Actual Environmental Samples.

Bacterial biosensors have great potential in contaminant detection for sensitivity, specificity, cost-effectiveness, and easy operation. However, the existing cadmium-responsive bacterial biosensors cannot meet the real-world detection requirements due to lack of sensitivity, specificity, and anti-interference capability. This study aimed to develop a bacterial biosensor for detecting the total and extractable cadmium in actual environmental samples. We constructed the cadmium-responsive biosensor with the regulatory element (cadmium resistance transcriptional regulatory, CadR) and the reporting element (GFP) and improved its performance by directed evolution. The mutant libraries of biosensors were generated by error-prone PCR and screened by continuous five-round fluorescence-activated cell sorting (FACS), and a bacteria variant epCadR5 with higher performance was finally isolated. Biosensor fluorescence intensity was measured by a microplate reader, and results showed that the evolved cadmium-responsive bacterial biosensor was of high sensitivity and specificity in detecting trace cadmium, with a detection limit of 0.45 µg/L, which is 6.8 times more specific to cadmium than that of the wild-type. Furthermore, microscopic qualitative analysis results showed that the bacteria could produce fluorescence response in a cadmium-contaminated soil matrix, and quantitative analysis results showed that the values of cadmium from epCadR5 bacteria were close to that from inductively coupled plasma-mass spectrometry. These results suggest that the biosensor may have a broad application prospect in the detection of cadmium-contaminated soil and water.

Advances in the research of osteosarcoma stem cells and its related genes.

Osteosarcoma is a malignant tumor that often occurs in adolescents. There is an urgent need for new treatment options for osteosarcoma due to its poor prognosis after metastasis. Cancer stem cell (CSC) theory states that CSCs represent a small proportion of cancer cells. These CSC have self-renewal ability and are closely associated with cancer growth and metastasis as well as chemotherapy resistance. Similarly, osteosarcoma stem cells (OSCs) play an important role in the growth, metastasis, and chemotherapy resistance of osteosarcoma cells. Targeting OSCs may represent a future treatment of osteosarcoma. Furthermore, some genes have been shown to regulate the growth, metastasis, and chemotherapy resistance of osteosarcoma cells by altering the stemness of OSCs. Targeting these genes may help in the treatment of osteosarcoma. This review mainly discusses recent advances in the research of OSCs and their related genes.

Family sex education and associated factors among preschoolers in Wuhu / 中国学校卫生

Objective@#To understand the status of family sex education and associated factors among preschoolers in Wuhu, providing scientific basis for child family sex education.@*Methods@#From July to August 2020, random cluster sampling was used to select 481 children from 5 kindergartens in Wuhu. A questionnaire survey among parents of these preschoolers regarding family sex education status and its influencing factors.@*Results@#A total of 285(59.25%) parents reported family sex education for children, and 196(40.75%) did not practice family sex education for their children. Multivariate Logistic regression analysis showed parental awareness of sex education content(OR=3.06, 95%CI=1.95-4.78), parental anxiety for child sexual assault (OR=1.82, 95%CI=1.11-2.99) were associated with higher rate of family sex education.@*Conclusion@#Family sex education among preschoolers in Wuhu should be further promoted. Sex education training towards parents might help improve children s family sex education.

miR-1225-5p Functions as a Tumor Suppressor in Osteosarcoma by Targeting Sox9.

microRNA plays an important role in the development of tumors, including osteosarcoma. However, the role of miR-1225-5p in osteosarcoma is currently unclear. First, we found that miR-1225-5p was downregulated in osteosarcoma cells relative to its levels in normal bone tissue by analyzing GSE28423 data in the GEO database. Using GSE39040, we found that low miR-1225-5p expression is associated with poor prognosis in patients with osteosarcoma, and we also found low miR-1225-5p expression in patients with recurrent osteosarcoma. We later demonstrated that osteosarcoma cell lines transfected with miR-1225-5p mimic had decreased ability to proliferate, migrate, and invade relative to control cells. To elucidate the mechanism by which miR-1225-5p inhibits the development of osteosarcoma, we identified Sox9 as a target gene of miR-1225-5p using the TargetScan website. We confirmed that Sox9 is the target gene of miR-1225-5p using the luciferase reporter assay. We then found that Sox9 is highly expressed in osteosarcoma by analyzing the GSE16088 and GSE42352 datasets and that high expression of Sox9 is associated with poor prognosis in patients with osteosarcoma using the R2 database. Further analysis using the TARGET database uncovered that high Sox9 expression is associated with a high recurrence rate in patients with osteosarcoma. Transfection of Sox9 siRNA inhibited the proliferation, migration, and invasiveness of osteosarcoma cells. We transfected miR-1225-5p together with Sox9 siRNA into osteosarcoma cells, observing strong inhibition of proliferation, migration, and invasiveness. Finally, exogenous expression of Sox9 partially reversed the anticancer effects of miR-1225-5p in osteosarcoma cells. Taken together, our findings suggest that miR-1225-5p functions as a tumor suppressor in osteosarcoma by targeting Sox9, thereby revealing new therapeutic targets for osteosarcoma.

Kyphosis correction and height restoration effects of percutaneous vertebroplasty.

BACKGROUND AND PURPOSE: Percutaneous vertebroplasty is known for its pain-relieving effect. Our purpose was to evaluate its effect on the kyphosis angle, wedge angle, and height of the fractured vertebral body. METHODS: We reviewed digital radiographs of 73 vertebral bodies in 53 patients before and after vertebroplasty. We measured the spinal kyphosis angle and the wedge angle of the fractured vertebral body. Ratios of the height of the anterior border, center, and posterior borders of the collapsed vertebra to the height at the posterior border of an adjacent normal vertebral body were measured. Gain from vertebroplasty and the restoration percentage (gain divided by loss) were calculated for each parameter. RESULTS: The kyphosis angle, wedge angle, anterior height, center height, and posterior height significantly improved after vertebroplasty. The mean reduction in the kyphosis angle was 4.3 degrees, and the wedge-angle reduction was 7.4 degrees. The mean wedge-angle reduction in fractured vertebral bodies containing gas was 10.2 degrees. Restoration percentages for the kyphosis angle and wedge angle were 19% and 44%, respectively. Gain in the height of the fractured vertebral bodies was 16.7% for the anterior border, 14% for the center, and 7% for the posterior border. Restoration percentages for the height of the vertebral body were 29% for the anterior border and 27% for the center. CONCLUSION: Vertebroplasty increases the height of the fractured vertebra and reduces the wedge and kyphosis angles. These effects are most remarkable in fractured vertebra containing gas.