Inhibition of GRK2 ameliorates the pristane-induced mouse SLE model by suppressing plasma cells differentiation.

Systemic lupus erythematosus (SLE) is a multifaceted autoimmune disorder characterized by diverse clinical manifestations and organ damage. Despite its elusive etiology, dysregulated subsets and functions of B cells are pivotal in SLE pathogenesis. Peoniflorin-6'-O-benzene sulfonate (CP-25), an esterification modification of Paeoniflorin, exhibits potent anti-inflammatory and immunomodulatory properties in autoimmune diseases (AID). However, the involvement of CP-25 and its target, GRK2, in SLE development has not been explored. In this study, we demonstrate that both genetic deficiency and pharmacological inhibition of GRK2 attenuate autoantibodies production, reduce systemic inflammation, and mitigate histopathological alterations in the spleen and kidney in the pristane-induced mouse SLE model. Importantly, our findings highlight that both genetic deficiency and pharmacological inhibition of GRK2 suppress plasma cells generation and restore dysregulated B-cell subsets by modulating two crucial transcription factors, Blimp1 and IRF4. Collectively, targeting GRK2 with CP-25 emerges as a promising therapeutic approach for SLE.

Therapeutic efficacy of drilling drainage combined with intraoperative middle meningeal artery occlusion in the management of chronic subdural hematoma: a clinical study.

BACKGROUND: The bone holes in the skull during surgical drainage were accurately located at the site of the MMA. The MMA was severed, and the hematoma was removed intraoperatively; furthermore, surgical drainage removed the pathogenic factors of CSDH. This study aimed to describe and compare the results of the new treatment with those of traditional surgical drainage, and to investigate the relevance of this approach. METHODS: From December 2021 to June 2023, 72 patients were randomly assigned to the observation group and the control group. The control group was treated with traditional surgical drainage, while the observation group was treated with DSA imaging to accurately locate the bone holes drilled in the skull on the MMA trunk before traditional surgical drainage. The MMA trunk was severed during the surgical drainage of the hematoma. The recurrence rate, time of indwelling drainage tube, complications, mRS, and other indicators of the two groups were compared, and the changes of cytokine components and imaging characteristics of the patients were collected and analyzed. RESULTS: Overall, 27 patients with 29-side hematoma in the observation group and 45 patients with 48-side hematoma in the control group were included in the study. The recurrence rate was 0/29 in the observation group and 4/48 in the control group, indicating that the recurrence rate in the observation group was lower than in the control group (P = .048). The mean indwelling time of the drainage tube in the observation group was 2.04 ± 0.61 days, and that in the control group was 2.48 ± 0.61 days. The indwelling time of the drainage tube in the observation group was shorter than in the control group (P = .003). No surgical complications were observed in the observation group or the control group. The differences in mRS scores before and after operation between the observation group and the control group were statistically significant (P < .001). The concentrations of cytokine IL6/IL8/IL10/VEGF in the hematoma fluid of the observation and control groups were significantly higher than those in venous blood (P < .001). After intraoperative irrigation and drainage, the concentrations of cytokines (IL6/IL8/IL10/VEGF) in the subdural hematoma fluid were significantly lower than they were preoperatively. In the observation group, the number of MMA on the hematoma side (11/29) before STA development was higher than that on the non-hematoma side (1/25), and the difference was statistically significant (P = .003). CONCLUSION: In patients with CSDH, accurately locating the MMA during surgical trepanation and drainage, severing the MMA during drainage, and properly draining the hematoma, can reduce the recurrence rate and retention time of drainage tubes, thereby significantly improving the postoperative mRS Score without increasing surgical complications.

Anti-arthritis effect of berberine associated with regulating energy metabolism of macrophages through AMPK/ HIF-1α pathway.

Berberine (BBR) is the effective constituent of Cortex phellodendri and was characterized as an excellent anti-microbial agent with significant anti-inflammatory effects. Previously, we had demonstrated that BBR alleviated the inflammatory response in adjuvant-induced arthritis (AA) rats by regulating polarization of macrophages. However, the exact mechanics by which BBR regulates macrophage polarization remained unclear. Here, we showed that BBR treatment had little influence on total number of macrophages in joints of AA rats, but increased the proportion of M2 macrophages and decreased the proportion of M1 macrophages. Meanwhile, we found BBR up-regulated the expression of AMP-activated protein kinase phosphorylation (p-AMPK) and down-regulated the expression of Hypoxia inducible factor 1α (HIF-1α) in synovial macrophages of AA rats. In vitro, using LPS-stimulated peritoneal macrophages from normal rats, we also verified that pretreatment with BBR promoted transition from M1 to M2 by up-regulating the expression of p-AMPK and suppressing the expression of HIF-1α. Compound C (an AMPK inhibitor) could abrogate the inhibition of BBR on migration of macrophages. Glycolysis of M1 suppressed by BBR through decreasing lactate export, glucose consumption, and increasing intracellular ATP content, which was remarkably reversed by Compound C. These findings indicated that anti-arthritis effect of BBR is associated with regulating energy metabolism of macrophages through AMPK/HIF-1α pathway.

[Chloroquine enhances cisplatin-induced apoptosis of nasopharyngeal carcinoma cells by inhibiting autophagy via upregulating miR129].

OBJECTIVE: To investigate the role of miR129 in mediating the effect of chloroquine to enhance cisplatin- induced apoptosis in nasopharyngeal carcinoma cells (HNE1). METHODS: MTT assay was used to detect the viability of HNE1 cells treated with different concentrations of cisplatin. Colony formation of HNE1 cells treated with cisplatin and chloroquine, alone or in combination, was observed using crystal violet staining. BALB/C unde mice were inoculated with HNE1 cells and randomly divided into 4 groups with 6 mice in each group. The mice received intraperitoneal injections of cisplatin and chloroquine, alone or in combination once every 3 days for 4 consecutive weeks, and the tumor growth was observed in each group. The expression of miR129 in HNE1 cells treated with chloroquine, cisplatin, or both was detected with qPCR. The effects of miR129 suppression with a miR129 inhibitor on the expressions of autophagy related proteins p62, LC3B, Beclin1 and the drug-resistant related protein P-glycoprotein (P-gp) were examined using Western blotting in HNE1 cells treated with chloroquine, cisplatin, or both; the changes in cell apoptosis were detected Annexin V/PI double staining. RESULTS: Chloroquine combined with cisplatin significantly inhibited HNE1 cell proliferation in vitro and the growth of HNE1 cell-derived tumor in nude mice as compared with cisplatin alone (P < 0.01). In cultured HNE1 cells, inhibition of the expression of miR129 significantly promoted autophagy and up-regulated P-gp expression (P < 0.01); Chloroquine obviously inhibited cisplatin-induced autophagy and up-regulated the expression of miR129 in HNE1 cells (P < 0.01). Transfection of the cells with the miR129 inhibitor abolished the inhibitory effect of chloroquine on cisplatin-induced autophagy, and significantly increased the cell survival rate (P < 0.05) and lower the cell apoptotic rate (P < 0.01) after combined treatment with chloroquine and cisplatin. CONCLUSIONS: Chloroquine enhances the pro-apoptotic effect of cisplatin by up-regulating miR129 to inhibit autophagy and drug resistance in HNE1 cells.

Using Micromachined Molds, Partial-curing PDMS Bonding Technique, and Multiple Casting to Create Hybrid Microfluidic Chip for Microlens Array.

In a previous study, we presented a novel manufacturing process for the creation of 6 × 6 and 8 × 8 microlens arrays (MLAs) comprising lenses with diameters of 1000 µm, 500 µm, and 200 µm within an area that covers 10 mm × 10 mm. In the current study, we revised the manufacturing process to allow for the fabrication of MLAs of far higher density (15 × 15 and 29 × 29 within the same area). In this paper, we detail the revised manufacturing scheme, including the micromachining of molds, the partial-curing polydimethylsiloxane (PDMS) bonding used to fuse the glass substrate and PDMS, and the multi-step casting process. The primary challenges that are involved in creating MLAs of this density were ensuring uniform membrane thickness and preventing leakage between the PDMS and glass substrate. The experiment results demonstrated that the revised fabrication process is capable of producing high density arrays: Design I produced 15 × 15 MLAs with lens diameter of 0.5 mm and fill factor of 47.94%, while Design II produced 29 × 29 MLAs with lens diameter of 0.25 mm and fill factor of 40.87%. The partial-curing PDMS bonding system also proved to be effective in fusing PDMS with glass (maximum bonding strength of approximately six bars). Finally, the redesigned mold was used to create PDMS membranes of high thickness uniformity (coefficient of variance <0.07) and microlenses of high lens height uniformity (coefficient of variance <0.15).

Microfabricated microfluidic platforms for creating microlens array.

The paper presents a novel and economic manufacturing process for microlens arrays (MLAs). This process uses micromilling machining, PDMS casting, and hybrid bonding between a glass substrate and PDMS membrane to create a microfluidic chip which is used for manufacturing MLAs on a PDMS substrates. MLAs of various diameters were fabricated for experiments, including 1000 µm, 500 µm, and 200 µm. The sag height of the MLAs is easily adjusted by controlling the pressure inside the microchannel to deform the PDMS membrane. Multiple experiments were conducted to characterize the performance of MLAs, the results of which demonstrate: (1) this fabrication process is able to manufacture MLAs with various dimensions and the diameter of an MLAs is determined by the size of micromilling bit and cutting path; (2) the sag height and curvature of MLAs can be controlled by the PDMS membrane thickness and the hydraulic pressure inside the microchannel; (3) an optical system was built to investigate the uniformity of MLAs and the experiment results showed uniform focal length of MLAs; (4) the resulting MLAs magnify tiny objects and significantly enhance the fluorescence signal emitted from the microchannel.

Micromachining Microchannels on Cyclic Olefin Copolymer (COC) Substrates with the Taguchi Method.

Micromilling is a straightforward approach to the manufacture of polymer microfluidic devices for applications in chemistry and biology. This fabrication process reduces costs, provides a relatively simple user interface, and enables the fabrication of complex structures, which makes it ideal for the development of prototypes. In this study, we investigated the influence of micromilling parameters on the surface roughness of a cyclic olefin copolymer (COC) substrate. We then employed factor analysis to determine the optimal cutting conditions. The parameters used in all experiments were the spindle speed, the feed rate, and the depth of cut. Roughness was measured using a stylus profilometer. The lowest roughness was 0.173 µm at a spindle speed of 20,000 rpm, feed rate of 300 mm/min, and cut depth of 20 µm. Factor analysis revealed that the feed rate has the greatest impact on surface quality, whereas the depth of cut has the least impact.

Expression, purification and crystallization of pecan (Carya illinoinensis) vicilin.

Tree nuts are responsible for many cases of severe food allergies. The 7S seed storage protein vicilin has been identified as a food allergen in many kinds of tree nuts. The vicilin protein consists of an N-terminal low-complexity region with antimicrobial activity and a C-terminal domain that forms a trimeric structure that belongs to the cupin superfamily. In this study, vicilin from pecan (Carya illinoinensis) was isolated and was expressed in bacteria for the first time. The cupin structural core of the protein, residues 369-792, was purified by metal-affinity and gel-filtration chromatography to high purity. Vicilin crystals were obtained and the best crystal diffracted to 2.65 Šresolution in space group P212121.