Towards Efficient Risky Driving Detection: A Benchmark and a Semi-Supervised Model.

Risky driving is a major factor in traffic incidents, necessitating constant monitoring and prevention through Intelligent Transportation Systems (ITS). Despite recent progress, a lack of suitable data for detecting risky driving in traffic surveillance settings remains a significant challenge. To address this issue, Bayonet-Drivers, a pioneering benchmark for risky driving detection, is proposed. The unique challenge posed by Bayonet-Drivers arises from the nature of the original data obtained from intelligent monitoring and recording systems, rather than in-vehicle cameras. Bayonet-Drivers encompasses a broad spectrum of challenging scenarios, thereby enhancing the resilience and generalizability of algorithms for detecting risky driving. Further, to address the scarcity of labeled data without compromising detection accuracy, a novel semi-supervised network architecture, named DGMB-Net, is proposed. Within DGMB-Net, an enhanced semi-supervised method founded on a teacher-student model is introduced, aiming at bypassing the time-consuming and labor-intensive tasks associated with data labeling. Additionally, DGMB-Net has engineered an Adaptive Perceptual Learning (APL) Module and a Hierarchical Feature Pyramid Network (HFPN) to amplify spatial perception capabilities and amalgamate features at varying scales and levels, thus boosting detection precision. Extensive experiments on widely utilized datasets, including the State Farm dataset and Bayonet-Drivers, demonstrated the remarkable performance of the proposed DGMB-Net.

Thermodynamic and Kinetic Modulation of Microfluidic Interfaces by DNA Nanoassembly Mediated Merit-Complementary Heteromultivalency.

The multivalent effect is often used to engineer microfluidic affinity interfaces to improve the target separation efficiency. Currently, no design rules exist for thermodynamic and kinetic tuning of properly joining multiple ligands. Herein, we developed a thermodynamic and kinetic modulating strategy of the microfluidic affinity interface via a merit-complementary-heteromultivalent aptamers functionalized DNA nanoassembly. Our strategy is built on the two types of identified aptamers that bind to distinct sites of EpCAM. The aptamer binding of one type is more rapid but less tight, while the other is opposite. By assembling the two types of aptamers together with a tetrahedral DNA framework, we fully exploited these aptamers' merits for tight and rapid recognition of EpCAM, leading to target cell capture with high efficiency and throughput. Our strategy provides a perspective on engineering multivalent recognition molecules through thermodynamic and kinetic tuning.

Auto-Panning: a highly integrated and automated biopanning platform for peptide screening.

Biopanning, a common affinity selection approach in phage display, has evolved numerous ligands for diagnosis, imaging, delivery, and therapy applications. However, traditional biopanning has suffered from time-consuming processes, highly-repetitive procedures and labor-intensive manual operation. Herein, a highly integrated and automated biopanning platform (Auto-Panning) is proposed. Based on digital microfluidics (DMF), biopanning processes are integrated on a chip with highly reproducible, precise, automated liquid manipulation. Therefore, 3 rounds of Auto-Panning can be accomplished within 16 h, instead of nearly a week of complicated manual operations. Auto-Panning has been used to evolve a specific peptide against cancer biomarker EphA2 with excellent cellular penetrating ability and significant invasion suppression biofunction, successfully demonstrating the practicality of the platform. Overall, as an automated programmable molecular screening platform, Auto-Panning will further promote the discovery and applications of novel ligands.

Highly paralleled emulsion droplets for efficient isolation, amplification, and screening of cancer biomarker binding phages.

Based on the linkage of genotype and phenotype, display technology has been widely used to generate specific ligands for profiling, imaging, diagnosis and therapy applications. However, due to the lack of effective monoclonal manipulation and affinity evaluation methods, traditional display technology has to undergo tedious steps of selection, clone isolation, amplification, sequencing, synthesis and characterization to obtain the binding sequences. To directly acquire high-affinity clones, we propose a double monoclonal display approach (dm-Display) for peptide screening based on highly paralleled monoclonal manipulation in emulsion droplets. dm-Display can monoclonally link the genotype, phenotype and affinity to realize integrated monoclonal separation, amplification, recognition and staining in one droplet so that discrete high-affinity clones can be quickly extracted. Monoclonal manipulations highly-parallelly occur in millions of droplets so that molecular screening of a highly diverse phage library is achieved. We have screened specific peptide ligands against CD71 and GPC1, proving the feasibility and generality of dm-Display. As a highly efficient ligand screening platform, dm-Display will promote the further development of molecular screening.

Selection and applications of functional nucleic acids for infectious disease detection and prevention.

Infectious diseases caused by pathogenic microorganisms such as viruses and bacteria pose a great threat to human health. Although a significant progress has been obtained in the diagnosis and prevention of infectious diseases, it still remains challenging to develop rapid and cost-effective detection approaches and overcome the side effects of therapeutic agents and pathogen resistance. Functional nucleic acids (FNAs), especially the most widely used aptamers and DNAzymes, hold the advantages of high stability and flexible design, which make them ideal molecular recognition tools for bacteria and viruses, as well as potential therapeutic drugs for infectious diseases. This review summarizes important advances in the selection and detection of bacterial- and virus-associated FNAs, along with their potential prevention ability of infectious disease in recent years. Finally, the challenges and future development directions are concluded.

Chlorin e6-loaded sonosensitive magnetic nanoliposomes conjugated with the magnetic field for enhancing anti-tumor effect of sonodynamic therapy.

In sonodynamic therapy (SDT), when Chlorin e6 (Ce6) accumulates in tumor tissues, its anti-tumor effect can be achieved by ultrasound activation. To increase the local drug concentration of Ce6 in tumor cells, we had established a novel drug delivery system, Ce6-loaded sonosensitive magnetic nanoliposome (Ce6/SML), which realized the targeting delivery by the external magnetic field. It was worth mentioning that the targeting release of Ce6/SML and the activation on Ce6 could be achieved simultaneously by ultrasound of SDT. In our study, after Ce6 was loaded into the sonosensitive magnetic nanoliposome (SML), the values of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in vitro and in vivo were determined, indicating the activation on Ce6 of ultrasound. The delivery system also displayed the tumor-targeting ability and anti-tumor activity, which associated with the determined tumor growth and expression levels of angiogenin (ANG), vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF-α). In conclusion, the Ce6/SML-SDT-Targeted delivery system could effectively enhance the anti-tumor activity of SDT and had a great potential application for the treatment of malignant tumors located in deep tissues.

Hydroxycamptothecin liposomes based on thermal and magnetic dual-responsive system: preparation, in vitro and in vivo antitumor activity, microdialysis-based tumor pharmacokinetics.

Due to the absence of lactone form of hydroxycamptothecin, the commercially available hydroxycamptothecin injection exhibits inefficient therapeutic effects. In this study, we constructed a novel delivery system (thermosensitive magnetic liposomes) that protects lactone form of hydroxycamptothecin from blood or water. After hydroxycamptothecin was loaded into the thermosensitive magnetic liposome (HCPT/TML), its in vitro and in vivo antitumor activity and microdialysis-based tumour pharmacokinetics were determined. The results demonstrated that HCPT/TMLs possessed favourable physicochemical features and significant cytotoxicity against the Huh-7 cells in vitro. In the in vivo antitumor study and tumour pharmacokinetics, HCPT/TMLs displayed effective targeting delivery and antitumor effects, which corresponded to the determined hydroxycamptothecin concentration in tumour tissue. In conclusion, this thermal and magnetic dual-responsive system can efficiently deliver hydroxycamptothecin to tumour tissue and has great potential application in cancer treatment.

An LC/MS quantitative and microdialysis method for cyclovirobuxine D pharmacokinetics in rat plasma and brain: The pharmacokinetic comparison of three different drug delivery routes.

To explore the brain-targeting of cyclovirobuxine D(CVB-D) after administered intranasally, the pharmacokinetics of CVB-D via three different drug delivery routes: intragastric (i.g.), intranasal (i.n.), and intravenous (i.v.) in rat brain and blood was compared. Firstly, an in vivo microdialysis method for sampling CVB-D in both plasma and brain of the rat was established. Secondly, a liquid chromatography-tandem mass spectrometry method has been developed and validated for determination of CVB-D in microdialysis samples. For plasma and brain microdialysis samples, liquid-liquid extraction was used and donepezil was chosen as internal standard. Both were followed by HPLC separation and positive electrospray ionization tandem mass spectrometry detection (ESI-MS/MS). Chromatographic separation was achieved on a agilent C18 column with a mobile phase of methanol-water (50:50, v/v) (pH 3.2) containing 0.1% formic acid and 5mM ammonium acetate. Mass spectrometric detection in the positive ion mode was carried out by selected reaction monitoring (MRM) of the transitions at m/z 403.4→372.3 for CVB-D and m/z 380.2→243.1 for donepezil (IS). Good linearities were obtained in the range of 10-4000ng/mL in rat microdialysates for CVB-D. The lowest limit of quantitation was 5ng/mL, with an extraction recovery >75%, and no significant matrix effects. Intra- and inter-day precisions were all <15% with accuracies of 97.26-116.20%. All of which proved that the established method was successfully applied to the pharmacokinetic study of CVB-D. Simultaneously, brain uptake and pharmacokinetic studies were performed by determination of CVB-D concentration in blood and brain respectively for CVB-D i.g., i.n. and i.v.. Results showed that the intranasal CVB-D could improve brain targeting and had advantages for direct nose to brain transport of CVB-D when compared with injection and oral delivery routes, which indicates that intranasal administration of CVB-D could be a promising approach for the treatment of cerebrovascular disease.

[Study on in vitro microdialysis recovery of hydroxycamptothecine].

OBJECTIVE: To establish the mothod to dectect the microdialysis recovery of HCPT and to investigate the influencing factors, thus to supply experimental basis for in-vivo microdialysis of HCPT. METHOD: The in vitro recovery of HCPT was detected by concentration difference method (increment method and decrement method). The influence of flow rates, medium concentration and temperature on the HCPT recovery and the stability were studied. RESULT: The recovery detected by increment method was the same as by decrement method. The recovery was independent of HCPT concentrations in the medium. The hydroxycamptothecine recovery had good stability and increased as the temperature rose. CONCLUSION: Microdialysis sampling can be used for the pharmacokinetic study of HCPT. Retrodialysis can be used for the determination of the HCPT in vivo recovery.

[Pharmacokinetics of nicotine in blood and brain using microdialysis and stable labelled isotope].

OBJECTIVE: Using the stable isotopes as the internal standard of microdialysis technology to establish a new method to study the whole and local brain dynamics of nicotine percutaneous preparations. METHOD: Using th healthy rats as experimental animals, administrating nicotine in abdominal transdermal way, then sample in the blood and brain simultaneously by microdialysis which use deuterium nicotine (DL-nicotine) as internal standard. Detecting the samples by LC-MS/MS method. RESULT: The configuration process in blood and brain both conforms to 2 compartments model, t1/2 is 29.38 min, t1/2beta is 208.51 min, AUC(0-infinity) is 152 127.10 microg x min x L(-1) in the blood t1/2 is 86.64 min, t1/2beta is 386.00 min, AUC(0-infinity) is 152 820.90 microg x min x L(-1) in the brain. CONCLUSION: Dl-nicotine can be used as internal standard of nicotine to correcte the recovery; Stable isotopes internal standard microdialysis technology can be used for studing the whole and the local pharmacokinetic of nicotine and also provide new ideas and methods to studing the process of new drug delivery system.

[Study on in vitro microdialysis recovery of Shuanghuanglian].

OBJECTIVE: To explore the feasibility of microdialysis techniqiue to be used in pharmacokinetic study of Chinese medicine, taking Shuanghuanglian as a model drug. METHOD: The samples were obtained by retrodialysis, determined by HPLC gradient elution to calculate the in vitro recovery rate (RR) of specific components. To study the difference of RR of a certain component in different dialysis mediums, and the effect of flow rates and concentration on RR. RESULT: Along with the increase of number of substances in the dialysis medium, the RR of specific components reduced. But the RR was independent of the concentration of the component in the dialysis medium. The RR reduced with the increasing flow rate in the same dialysis medium. CONCLUSION: The microdialysis technique can be used in pharmacokinetics study of Chinese medicine.

[Study of pharmacokinetics of nicotine in local brain by using microdialysis and stable labeled isotope].

The paper is to report the study of pharmacokinetics of transdermal administered nicotine in the brain of freely moving rat by using microdialysis with stable labeled isotope as internal standard. The pharmacokinetic behavior of nicotine in Sprague Dawley rat brain was investigated after intranasal administration (3.75 mg). Brain fluid samples were collected by intracerebral microdialysis with DL-nicotine as internal standard. Concentrations of nicotine and DL-nicotine in the sample were measured by HPLC-MS/MS. Main pharmacokinetic parameters were calculated and analyzed by Das 2.0 pharmacokinetic software. The recovery of nicotine and the delivery of DL-nicotine were the same. The fate of absorption and distribution was two compartment model and the values of t1/2alpha was 170.31 min, t1/2beta was 263.30 min and the AUC(0-infinity) was 2.75 x 10(5) microg x L(-1) min separately. DL-nicotine can be used to calibrate the recovery of nicotine, and the new method of stable isotope microdialysis can be used to study the pharmacokinetics of freely moving rat. It will make sense for the treatment of addiction of tobacco and provide a new thought for the research of pharmacokinetics-pharmacodynamic combination.

[Internal standard method to determine the recovery of nicotine in microdialysis].

The paper reports the evaluation of the feasibility of using internal standard method for the determination of nicotine recovery in microdialysis in vitro. This in vitro experiment included two conditions. Nicotine and codeine phosphate were dissolved in Ringer's solution. Nicotine, codeine phosphate and the mixture of them were perfused through the CMA30 linear probe separately to calculate the proportion of the recovery (or delivery) of nicotine to that of codeine phosphate. And then codeine was perfused through the probe which was immersed in nicotine solution with different concentrations to calculate the proportion, too. In another condition nicotine was dissolved in rat plasma. The rat plasma protein binding rate was determined by using retrodialysis and internal standard method in vitro. The results are as follows: the proportion of the recovery (or delivery) of nicotine to that of codeine phosphate was fairly stable. The delivery of codeine was independent of nicotine concentration in the external medium. Protein binding rate determined by retrodialysis was almost the same as that determined by internal standard method. It suggests that the internal standard method is an effective way in the determination of nicotine recovery and codeine phosphate can be used as the internal standard.

[Study on optimization of formulation and preparation process of sinomenine liposomes].

OBJECTIVE: To optimize the formulation and preparation process of sinomenine liposomes. METHOD: Method of aether injection and mixture uniform design were adopted to determine the formulation of sinomenine liposomes is the proportion of phospholipids, cholesterol and Vitamin E with the index of entrapment efficiency. And the single-factor test was used to study the preparation process of the liposomes, including the volume of buffer solution, the preparation temperature and the ultrasonic time. RESULT: The optimized formulation was that the ratio of sinomenine : phospholipids : cholesterol : vitamin E mass ratio was 8.92 : 60.35 : 28.81 : 1.91. The volume of buffer solution was 50 mL x g(-1) membrane, the preparation temperature was 50 degrees C, and the ultrasonic time was 20 min. CONCLUSION: Satisfactory shape and entrapment efficiency of the liposomes can be obtained by the optimized formulation and preparation process.

[Dynamic determination of sinomenine in skin by microdialysis].

OBJECTIVE: To explore the feasibility and advantages of using microdialysis as sampling method for dynamic determination of sinomenine in topical skin. METHODS: In this study, sinomenine was administered to the rats by transdermal drug delivery system and celiac injection. With microdialysis technique for sampling, the concentration of sinomenine in dialysate was determined by high performance liquid chromatography (HPLC). RESULTS: Under existing determination condition, topical drug concentration in the skin of rats was hard to be detected after sinomenine administered to the skin, but it could be detected after celiac injection. CONCLUSION: Microdialysis sampling method can be used to determine topical drug concentration in the skin dynamically, and this method is better than traditional methods obviously.

[Bioequivalence study of sinomenine patch by microdialysis].

OBJECTIVE: To study bioequivalence of Sinomenine patch made by different preparation process, and to testify feasibility and superiority of microdialysis as a new method in topical bioequivalence study. METHOD: Normal gel patch and liposome gel patch of sinomenine were prepared by different preparation, nude mouse served as the experimental subjects sampling method of drug in the skin was tissue homogenization microdialysis, and drug concentration in dialysate was determined by HPLC. RESULT: Results of tissue homogenization showed that liposome gel patch leads more remainder drug in the skin of nude mouse than normal gel patch, and results of microdialysis showed that liposome gel patch led higher instantaneous drug concentration than normal gel patch. Concentration-time curve of sinomenine in the skin accorded with the results of most dermal delivery systems studies over the world. CONCLUSION: Topical bioequivalence of liposome gel patch of sinomenine is higher than that of normal gel patch of sinomenine. Microdialysis can be used to study bioavailability and bioequivalence of different preparation.

[Study on determination of entrapment efficiency of sinomenine liposomes].

OBJECTIVE: To establish an HPLC method for the determination of entrapment efficiency of sinomenine liposomes. METHOD: The liposomes and dissociated drugs were separated by sephadex filtration, mini-column centrifugation and dialysis. The methodology study and the optimization of determining condition were carried out at the same time. RESULT: Sephadex filtration could effectively separate the sinomenine liposomes from dissociated sinomenine. The column recovery was 98.8%, the average entrapment efficiency of three tests was64.9%, RSD 2.67%. CONCLUSION: The method was simple, exact, and had a good reappearance. It can be used to examine the entrapment efficiency of sinomenine liposomes.

[Effect of penetration enhancers on the transdermal penetration of sinomenine liposome patch].

OBJECTIVE: To study the effect of penetration enhancers on the transdermal penetration of sinomenine liposome patch. METHODS: We firstly established the experimental method of transdermal penetration, and the methodology study was carried out at the same time. Furthermore, we added 5% Azone, 10% Oleic acid, 1% PG + 4% Azone, 2% PG + 8% Oleic acid into the patch. The cumulative penetration amounts were determined during the test of transdermal penetration and the part lagged in the skin was determined after the test. RESULTS: The cumulative penetration amounts released from the patch increased, while the part lagged in the skin decreased at the same time. CONCLUSIONS: The results indicate that penetration enhancers can facilitate drug through corneum into dermis, but can't enhance the drug' s accumulation in the corneum. It is concluded that penetration enhancers can destroy the two-molecule structure of liposomes and cause drug leakage, so penetration enhancers should not be added into liposome patch of part functions.