Stauntonia chinensis injection relieves neuropathic pain by increasing the expression of PSD-95 and reducing the proliferation of phagocytic microglia.

Neuroinflammation induced by engulfment of synapses by phagocytic microglia plays a crucial role in neuropathic pain. Stauntonia chinensis is extracted from Stauntonia chinensis DC, which has been used as a traditional Chinese medicine to control trigeminal neuralgia or sciatica. However, the specific anti-neuralgia mechanism of Stauntonia chinensis is unknown. In this study, the analgesic effect of Stauntonia chinensis injection (SCI) in mice with neuropathic pain and the possible mechanisms are explored. We find that a local injection of 0.1 mL Stauntonia chinensis for 14 days can considerably relieve mechanical hyperalgesia and thermal hyperalgesia in mice with sciatic chronic constriction injury (CCI). Immunofluorescence staining shows that SCI reduces neuroinflammation in the spinal cord of CCI mice. RNA sequencing reveals that the expression of postsynaptic density protein 95 (PSD-95), a postsynaptic scaffold protein, is downregulated in the spinal cord of CCI mice, but upregulated after SCI administration. Immunofluorescence experiments also demonstrate that SCI administration reverses microglia proliferation and PSD-95 downregulation in CCI mice. These data suggest that SCI relieves neuropathic pain by increasing the expression of PSD-95 and reducing the proliferation of phagocytic microglia.

The complement system: a potential target for the comorbidity of chronic pain and depression.

The mechanisms of the chronic pain and depression comorbidity have gained significant attention in recent years. The complement system, widely involved in central nervous system diseases and mediating non-specific immune mechanisms in the body, remains incompletely understood in its involvement in the comorbidity mechanisms of chronic pain and depression. This review aims to consolidate the findings from recent studies on the complement system in chronic pain and depression, proposing that it may serve as a promising shared therapeutic target for both conditions. Complement proteins C1q, C3, C5, as well as their cleavage products C3a and C5a, along with the associated receptors C3aR, CR3, and C5aR, are believed to have significant implications in the comorbid mechanism. The primary potential mechanisms encompass the involvement of the complement cascade C1q/C3-CR3 in the activation of microglia and synaptic pruning in the amygdala and hippocampus, the role of complement cascade C3/C3a-C3aR in the interaction between astrocytes and microglia, leading to synaptic pruning, and the C3a-C3aR axis and C5a-C5aR axis to trigger inflammation within the central nervous system. We focus on studies on the role of the complement system in the comorbid mechanisms of chronic pain and depression.

Genome-Wide Identification, Characterization, and Expression Analysis of NF-Y Gene Family in Ginkgo biloba Seedlings and GbNF-YA6 Involved in Heat-Stress Response and Tolerance.

Nuclear factor Y (NF-Y) transcription factors play an essential role in regulating plant growth, development, and stress responses. Despite extensive research on the NF-Y gene family across various species, the knowledge regarding the NF-Y family in Ginkgo biloba remains unknown. In this study, we identified a total of 25 NF-Y genes (seven GbNF-YAs, 12 GbNF-YBs, and six GbNF-YCs) in the G. biloba genome. We characterized the gene structure, conserved motifs, multiple sequence alignments, and phylogenetic relationships with other species (Populus and Arabidopsis). Additionally, we conducted a synteny analysis, which revealed the occurrence of segment duplicated NF-YAs and NF-YBs. The promoters of GbNF-Y genes contained cis-acting elements related to stress response, and miRNA-mRNA analysis showed that some GbNF-YAs with stress-related cis-elements could be targeted by the conserved miRNA169. The expression of GbNF-YA genes responded to drought, salt, and heat treatments, with GbNF-YA6 showing significant upregulation under heat and drought stress. Subcellular localization indicated that GbNF-YA6 was located in both the nucleus and the membrane. Overexpressing GbNF-YA6 in ginkgo callus significantly induced the expression of heat-shock factors (GbHSFs), and overexpressing GbNF-YA6 in transgenic Arabidopsis enhanced its heat tolerance. Additionally, Y2H assays demonstrated that GbNF-YA6 could interact with GbHSP at the protein level. Overall, our findings offer novel insights into the role of GbNF-YA in enhancing abiotic stress tolerance and warrant further functional research of GbNF-Y genes.

Activation of locus coeruleus-spinal cord noradrenergic neurons alleviates neuropathic pain in mice via reducing neuroinflammation from astrocytes and microglia in spinal dorsal horn.

BACKGROUND: The noradrenergic neurons of locus coeruleus (LC) project to the spinal dorsal horn (SDH), and release norepinephrine (NE) to inhibit pain transmission. However, its effect on pathological pain and the cellular mechanism in the SDH remains unclear. This study aimed to explore the analgesic effects and the anti-neuroinflammation mechanism of LC-spinal cord noradrenergic pathway (LC:SC) in neuropathic pain (NP) mice with sciatic chronic constriction injury. METHODS: The Designer Receptors Exclusively Activated by Designer Drugs (DREADD) was used to selectively activate LC:SC. Noradrenergic neuron-specific retro-adeno-associated virus was injected to the spinal cord. Pain threshold, LC and wide dynamic range (WDR) neuron firing, neuroinflammation (microglia and astrocyte activation, cytokine expression), and α2AR expression in SDH were evaluated. RESULTS: Activation of LC:SC with DREADD increased the mechanical and thermal nociceptive thresholds and reduced the WDR neuron firing. LC:SC activation (daily, 7 days) downregulated TNF-α and IL-1ß expression, upregulated IL-4 and IL-10 expression in SDH, and inhibited microglia and astrocytes activation in NP mice. Immunofluorescence double staining confirmed that LC:SC activation decreased the expression of cytokines in microglia of the SDH. In addition, the effects of LC:SC activation could be reversed by intrathecal injection of yohimbine. Immunofluorescence of SDH showed that NE receptor α2B-AR was highly expressed in microglia in CCI mice. CONCLUSION: These findings indicate that selective activation of LC:SC alleviates NP in mice by increasing the release of NE and reducing neuroinflammation of astrocytes and microglia in SDH.

A low-cost and high-efficiency method for four-inch silicon nano-mold by proximity UV exposure.

Nano-mold is an essential tool for nano-imprinting. However, large-area nano-mold fabrication relies on expensive equipment or complicated processing. Silicon nano-molds were achieved by proximity ultraviolet lithography and reactive ion etching (RIE). By optimizing the parameters in the processes of exposure, development, and RIE, silicon nano-mold with nano-scale ridges were fabricated with high-precision. The achieved minimum width of nano-ridges was 263 nm. This method is capable of fabricating silicon nano-mold covering four-inch wafer, which is simple, efficient and free from costly equipment.

Fabrication of 2D silicon nano-mold by side etch lift-off method.

Nano-imprint technology is a method of nano-pattern reproduction, has the characteristics of high resolution, high throughput, and low-cost. It can reduce the complexity and cost of the equipment while improving the resolution, which considered a promising industrial production technology. The key to nanoimprinting lies in the mold, and the quality of the mold directly determines the quality of the imprinted graphics. Here, a method for fabricating sub-100 nm concave 2D silicon nano-mold by side etch lift-off is proposed. The effects of different wet etching time and the metal deposition angle on the width of nanochannels were studied. The measurement result of dry etching shows that on the entire 4 inch silicon wafer, the width of the nanochannel varies by 4% and the depth by 2%. The width of the nanochannel between chips varies by 0.7%, and the depth variation is 1%. With this new method, high-precision and large-scale silicon nano-mold can be produced, which has great potential for realizing high-precision and low-cost manufacturing of nano devices.

Nano-Gaps Fabricated by Thermal Evaporation and Stripping Techniques.

The fabrication of inexpensive nano-gaps is vitally important for the research and application of nanochannel-based devices. This study presents a low-cost and simple method for the fabrication of nano-gaps using thermal evaporation and stripping techniques. The structural morphology of metal films deposited on the convex structures of photoresist by sputtering and thermal evaporation was studied. The effect of angles of thermal evaporation on the width of nano-gaps was investigated. The characteristics of metal film deposited on the convex structures of photoresist and spaces between these convex structures after stripping were investigated, and the adhesive force between the metal film and silicon substrate was also analyzed. Finally, a metal film of Cu was deposited on the convex structures of photoresist by thermal evaporation. After stripping, nano-gaps with a width of 187 nm were fabricated. The method proposed in this paper can be employed to mass-produce two-dimensional nanochannels based devices at low cost.

A Novel Room-Temperature Bonding Method Based on Electrohydrodynamic Printing.

Microfluidic chips made by traditional materials (glass and silicon) are still important for fluorescence tests, biocompatible experiments, and high temperature applications. However, the majority of the present bonding methods suffer from ultra-clean requirement, complicated fabrication process, and low production efficiency. In the present work, an Electrohydrodynamic printing assist bonding method was proposed. By this method, the ultraviolet-cured-glue dots were printed onto the silicon substrate, and then the patterned glass and silicon substrate can be bonded together at room temperature. The influence of printing condition (nozzle inner-diameter, applied voltage, printing height, and flow rate) on the diameter of printed dot was analyzed by experiments. By the optimized printing condition, the glass-silicon microfluidic chip can be well bonded. The bonding strength and leakage test demonstrated the high bonding quality of the microfluidic chip (bonding strength of 28 MPa and leakage pressure of 3.5 MPa).

The Fabrication of Polymethyl Methacrylate Nozzles for Electrohydrodynamic Printing.

Electrohydrodynamic (EHD) jet printing enables rapid prototyping high-resolution and low-cost lines with width of micrometer or even nanometer. However, EHD printing always suffers from nozzle clogging when the nozzle inner-diameter decrease to micro-scale. Thus fabrication of low cost nozzles becomes significantly important. In this work, 50 µm wide and 12.5 µm deep PMMA (Polymethyl Methacrylate) nozzles were fabricated without using traditional expensive glass capillary pulling approach. To replicate PMMA nozzle with high precision, the embossing condition was optimized according to replication precision, the deformation rate, and maximum stress. To nearly fully bond PMMA nozzle with intact PMMA microchannel, the bonding condition was optimized according the bonding rate and dimension loss of PMMA microchannel. The availability of the fabricated PMMA nozzle was finally verified by EHD printing experiments.

Effects of norepinephrine on microglial neuroinflammation and neuropathic pain.

Norepinephrine (NE) is an important neurotransmitter in the central nervous system. NE is released from locus coeruleus neurons and is involved in a variety of physiological and pathological processes. Neuroinflammation is a common manifestation of many kinds of neurological diseases. The activation of microglia directly affects the status of neuroinflammation. Several kinds of adrenergic receptors, which anchor on microglia and can be regulated by NE, affect the activation of microglia and neuroinflammation. NE influences chronic pain, anxiety, and depression by regulating the activation of microglia.

Nano-Patterns of Photoresist Fabricated by Ultraviolet Lithography Technology.

Nanofluidics devices with two-dimensional nano-structures have attracted extensive interests for biological and chemical applications. The fabrication of nanoscale mask patterns with controllable line width is an indispensable process for manufacturing two-dimensional nano-structures. However, a simple and low-cost method for fabricating two-dimensional nano-patterns is still a challenge. In this work, a novel simple and low-cost method for fabricating nanoscale mask patterns with controllable line width, based on the ultraviolet exposure is presented. In the experiment, a layer of AZ5214 reversal photoresist was exposed to the ultraviolet light through the photomask with microscale patterns. After the lithography, nanoscale photoresist mesas patterns instead of microscale patterns were produced. The photoresist mesas with 400~800 nm width were fabricated. Meanwhile, by adjusting the parameters of exposure time, various nanoscale dimensions of photoresist mesas can be obtained. The proposed method overcomes limitations of the conventional ultraviolet lithography technology without the need to the expensive nanolithography equipment. Our novel fabrication method should be a useful tool for two-dimensional nano-structures fabrication due to its advantages of simple fabrication process, well controllability, and low-cost.

Fast Microfluidic Chip Fabrication Technique by Laser Erosion and Sticky Tape Assist Bonding Technique.

Disposable microfluidic chips are becoming increasingly important for biological and chemical researches. Future advances in their commercial applications depends on the mass fabrication of low cost microfluidic chip. In this study we are presenting a simple, low cost and fast way of fabricating PMMA microfluidic chips based on laser erosion. The influence of the width and depth of PMMA microchannels on erosion current and erosion times was analyzed. To bond the open PMMA microchannels at low pressure and temperature, sticky tape assist bonding method was proposed. By this method, the microfluidic chip can be fully sealed without using any equipment. The leakage test indicated that the bonded microfluidic chip can endure an pressure up to 0.82 MPa.

Polycarbonate Nanofluidic Chip Fabrication Technique by Hot Embossing and Thermal Bonding.

Nanofluidic devices are becoming increasingly critical for medical, chemical and biological applications. In this paper, a 2D Polycarbonate (PC) nanofluidic chip was fabricated by hot embossing and thermal bonding method. In this paper, the effect of the hot embossing parameters on the replication precision of 2D PC nanochannels was investigated by finite element method. To increase the bonding rate of the chip, the parameters of the thermal bonding process were optimized. Under the optimized parameters the 2D PC nanofluidic chip was successfully fabricated. The results show that the replication precision of the nanochannels can be as high as 97% and the bonding rate of the chip can be 96%. The fluorescent images demonstrate that there is no block and leakage through the whole microchannels and nanochannels. It is expected that this fabrication method has great potential to fabricate 2D nanofluidic chip with low cost and high precision.

Low auto-fluorescence fabrication methods for plastic nanoslits.

Plastic nanofluidic devices are becoming increasingly important for biological and chemical applications. However, they suffer from high auto-fluorescence when used for on-chip optical detection. In this study, the auto-fluorescence problem of plastic nanofluidic devices was remedied by newly developed fabrication methods that minimise their auto-fluorescence: one by depositing a gold (Au) layer on them, the other by making them ultra-thin. In the first method, the Au layer [minimum thickness is 40 nm on 150 µm SU-8, 50 nm on 1 mm polyethylene terephthalate (PET), and 40 on 2 nm polymethyl methacrylate (PMMA)] blocks the auto-fluorescence of the polymer; in the second method, auto-fluorescence is minimised by making the chips ultra-thin, selected operating thickness of SU-8 is 20 µm, for PET it is 150 µm, and for PMMA it is 0.8 mm.

A novel 2D silicon nano-mold fabrication technique for linear nanochannels over a 4 inch diameter substrate.

A novel low-cost 2D silicon nano-mold fabrication technique was developed based on Cu inclined-deposition and Ar(+) (argon ion) etching. With this technique, sub-100 nm 2D (two dimensional) nano-channels can be etched economically over the whole area of a 4 inch n-type <100> silicon wafer. The fabricating process consists of only 4 steps, UV (Ultraviolet) lithography, inclined Cu deposition, Ar(+) sputter etching, and photoresist &Cu removing. During this nano-mold fabrication process, we investigated the influence of the deposition angle on the width of the nano-channels and the effect of Ar(+) etching time on their depth. Post-etching measurements showed the accuracy of the nanochannels over the whole area: the variation in width is 10%, in depth it is 11%. However, post-etching measurements also showed the accuracy of the nanochannels between chips: the variation in width is 2%, in depth it is 5%. With this newly developed technology, low-cost and large scale 2D nano-molds can be fabricated, which allows commercial manufacturing of nano-components over large areas.

Two dimensional PMMA nanofluidic device fabricated by hot embossing and oxygen plasma assisted thermal bonding methods.

A method for obtaining a low-cost and high-replication precision two-dimensional (2D) nanofluidic device with a polymethyl methacrylate (PMMA) sheet is proposed. To improve the replication precision of the 2D PMMA nanochannels during the hot embossing process, the deformation of the PMMA sheet was analyzed by a numerical simulation method. The constants of the generalized Maxwell model used in the numerical simulation were calculated by experimental compressive creep curves based on previously established fitting formula. With optimized process parameters, 176 nm-wide and 180 nm-deep nanochannels were successfully replicated into the PMMA sheet with a replication precision of 98.2%. To thermal bond the 2D PMMA nanochannels with high bonding strength and low dimensional loss, the parameters of the oxygen plasma treatment and thermal bonding process were optimized. In order to measure the dimensional loss of 2D nanochannels after thermal bonding, a dimension loss evaluating method based on the nanoindentation experiments was proposed. According to the dimension loss evaluating method, the total dimensional loss of 2D nanochannels was 6 nm and 21 nm in width and depth, respectively. The tensile bonding strength of the 2D PMMA nanofluidic device was 0.57 MPa. The fluorescence images demonstrate that there was no blocking or leakage over the entire microchannels and nanochannels.

Design, simulation and evaluation of improved air amplifier incorporating an ion funnel for nano-ESI MS.

An improved air amplifier design that takes advantage of the combined effects of aerodynamic and electrodynamic focusing was developed to couple a nanoelectrospray ionisation (nano-ESI) source and the heated mass spectrometer inlet to improve the sensitivity of a mass spectrometer. The new design comprises an electrodynamic ion funnel integrated into the main air pathway of the air amplifier to more effectively focus and transmit gas-phase ions from the nano-ESI source into the heated mass spectrometer inlet. Numerical computational fluid dynamics simulations were carried out using a commercial software package, ANSYS FLUENT, to provide more detailed information about the device's performance. The gas flow field as well as the electric field patterns and the Lagrangian ion motion were conveniently simulated using this single package and custom-written, user-defined functions. Experimental results show a nearly five-fold improvement in reserpine ion intensity with the air amplifier operated at a nitrogen gauge pressure of 40 kPa and no direct current (DC) or radiofrequency (RF) potentials applied to the ion funnel when the distance between the electrospray emitter and sampling inlet tube was 24 mm, as compared to direct sample infusion from the same distance without the air amplifier. More importantly, a nearly three-fold additional gain in ion intensity was measured when both DC and RF potentials were co-applied, resulting in more than a 13-fold overall ion intensity gain which could be attributed to the combined air amplifier aerodynamic and ion funnel electrodynamic focusing effect.

A novel hybrid patterning technique for micro and nanochannel fabrication by integrating hot embossing and inverse UV photolithography.

Nanofluidic devices with micro and nanostructures are becoming increasingly important for biological and chemical applications. However, the majority of the present fabrication methods suffer from a low pattern transfer quality during the simultaneous embossing of the microscale and nanoscale patterns into a thermoplastic polymer due to insufficient polymer flow. In this work, a novel hybrid patterning technique, integrating hot embossing and inverse ultraviolet (UV) photolithography, is developed to fabricate micro and nanochannels with a high replication precision of the SU-8 layer. The influence of embossing temperature and time on the replication precision was investigated. The effect of UV lithography parameters on the micro and nanochannel pattern was analyzed. To improve the SU-8 bonding strength, the influence of the O2 plasma treatment parameters on the water contact angles of the exposed and unexposed SU-8 layer were studied. A complete SU-8 nanofluidic chip with 130 nm wide and 150 nm deep nanochannels was successfully fabricated with a replication precision of 99.5%. Compared with most of the current processing methods, this fabrication technique has great potential due to its low cost and high pattern transfer quality of the SU-8 micro and nanochannels.

Fabrication of two dimensional polyethylene terephthalate nanofluidic chip using hot embossing and thermal bonding technique.

We present in this paper a method for obtaining a low cost and high replication precision 2D (two dimensional) nanofluidic chip with a PET (polyethylene terephthalate) sheet, which uses hot embossing and a thermal bonding technique. The hot embossing process parameters were optimized by both experiments and the finite element method to improve the replication precision of the 2D nanochannels. With the optimized process parameters, 174.67 ± 4.51 nm wide and 179.00 ± 4.00 nm deep nanochannels were successfully replicated into the PET sheet with high replication precision of 98.4%. O2 plasma treatment was carried out before the bonding process to decrease the dimension loss and improve the bonding strength of the 2D nanofluidic chip. The bonding parameters were optimized by bonding rate of the nanofluidic chip. The experiment results show that the bonding strength of the 2D PET nanofluidic chip is 0.664 MPa, and the total dimension loss of 2D nanochannels is 4.34 ± 7.03 nm and 18.33 ± 9.52 nm, in width and depth, respectively. The fluorescence images demonstrate that there is no blocking or leakage over the entire micro- and nanochannels. With this fabrication technology, low cost polymer nanochannels can be fabricated, which allows for commercial manufacturing of nano-components.

Extraordinary optical transmission based on subwavelength metallic grating with ellipse walls.

This paper presents a nanometer-sized metallic film periodically pierced by narrow slits with ellipse walls deposited on a substrate that demonstrates special optical properties of broadband extraordinary optical transmission (BEOT). Compared to slits with straight walls, the metal slits with nonlinearly tapered ellipse walls can collect more light on the upper surface, which is coupled into a gap plasmon polariton propagating along the ellipse walls, then delivers the light at the smaller exit slit opening. In the visible spectral region, BEOT of TM-polarized light is achieved with up to 80% transmission at resonance, which is resulted from the simultaneous enhancement of zero-order slit resonance and higher-order slit resonances excited due to the existence of the substrate. The spectral range of BEOT is limited by Wood-Rayleigh anomalies and surface plasmon polariton resonances (SPPs). The BEOT spectrum of oblique incidence with small incident angle that is divided into two separate bands are also presented and analyzed theoretically. This metallic grating overcomes the low optical transmission limit of the structures with wavelength-sized grating period in visible and near-IR regions. It can be used to design nanostructured BEOT polarizer, which is an important component in novel biomimetic-based optoelectronic systems especially those in skylight polarized environment.