Enhancement of dissolution and oral bioavailability by adjusting microenvironment pH in crocetin ternary solid dispersions: Optimization, characterization, in vitro evaluation, and pharmacokinetics.

The most promising active ingredient of Crocus sativus L., crocetin (CCT), has been demonstrated to possess many biological activities. However, only a few studies have been conducted on CCT formulation, especially in oral formulation, mainly due to its insolubility in water, which limits its application for oral administration. This article reports an equilibrium saturation solubility and single-pass intestinal perfusion studies conducted to classify the biopharmaceutics classification system (BCS) of CCT. To enhance in vitro dissolution and in vivo oral bioavailability, ternary solid dispersions of CCT (CCT-SDs) with soluplus (SOL) as hydrophilic carrier and meglumine (MEG) as alkalizer were optimized using response surface methodology (RSM) with central composite design (CCD) experiments. Four different preparation methods were evaluated using the optimal formulation, including solvent evaporation, ball milling, spray drying, and freeze-drying. Prepared formulations were characterized by TG-DSC, FTIR, X-RPD, and SEM; the pharmacokinetic studies were performed in rats after oral administration. The cumulative dissolution rate of CCT-SDs containing SOL and MEG prepared by the ball milling method was 97.1% at 15 min and remained at 95.6% at 480 min, which was significantly higher than that of untreated CCT. The lower crystallinity, smaller particle size, and higher microenvironment pH (pHM) were observed in CCT-SDs prepared by the ball milling method. In vivo absorption of CCT-SDs (Cmax = 52.789 ± 12.441 µg/mL and AUC0-12 = 191.748 ± 35.043 µg/mL·h) was greater than untreated CCT (Cmax = 5.918 ± 1.388 µg/mL and AUC0-12 = 44.309 ± 7.264 µg/mL·h). In conclusion, the current study provides ternary solid dispersion formulation of CCT to increase the in vitro dissolution and in vivo bioavailability, which will benefit the commercial production and future clinical applications of CCT.

Effects of Lactobacillus acidophilus and L. reuteri on bone mass and gut microbiota in ovariectomized mice.

Lactobacillus acidophilus is widely used as a food additive or medication in our daily lives. The objective of this study was to investigate the effects of L. acidophilus and L. reuteri on bone mass in OVX mice and their associated mechanisms. Fifty 6-week-old female C57BL/6J mice were subjected to five different treatments: sham surgery, OVX surgery, OVXandL. reuteri fed, OVXandL. acidophilus fed, OVXandboth L. reuteri and L. acidophilus co-fed, respectively. Serum samples were collected, and IL-1ß,IL-6,TNF-α, and OCN levels were determined. The bone volume fraction and trabecular number, trabecular thickness, trabecular separation, and cortical thickness of the mice were analyzed by micro-CT in both femurs. Mice feces were taken for Illumina high-throughput sequencing to analyze the microbial composition and characteristics. After probiotic feeding, the bone volume fraction, the trabecular number, and the trabecular thickness increased, and the trabecular separation decreased in OVX mice. IL-1ß, IL-6, and TNF-α in the blood significantly decreased. The observed Chao1 and ACE indexes increased significantly. Changes in intestinal microorganisms occurred in all groups of mice. The change of index in the gut microbes, may indicate that the bone mass of OVX mice is changing. L. acidophilus shares the same role as L. reuteri in preventing bone loss in OVX mice. The mechanism of action may be through inhibition of the activation of inflammatory factors in the osteoclast activation pathway in bone metabolism, modulation of gut microbial diversity, and alteration of the richness of specific microorganisms that lead to attenuation of bone loss.

3D-Printed Bionic Titanium Alloy Artificial Lamina Prevents Epidural Adhesion and Restores the Stability After Laminectomy in Pigs.

Laminectomy can cause the dura mater to adhere to the surrounding scar tissue, leading to soft spinal stenosis after surgery. Although artificial laminae are considered ideal substitutes, they present challenges such as insecure fixation and insufficient bionics. In this study, we fabricated a bionic titanium alloy artificial lamina using three-dimensional (3D)-printing technology and evaluated its adhesion prevention and stability after laminectomy in pigs. An in vitro biomechanical pull-out resistance test indicated that the pull-out strength of the artificial lamina was close to that of a single pedicle screw and was significantly higher than that of a cortical screw. In vivo animal implantation results indicated precise laminectomy and artificial lamina implantation, as well as a safe operation process with the assistance of guide plates. X-ray and computed tomography results indicated the well fixation of bionic titanium alloy artificial lamina and screws 10 weeks after laminectomy. The artificial lamina was not loosened after being removed from pigs (postoperative week 12), exhibiting good stability. Additionally, no adhesion was observed in the artificial lamina group, whereas a large amount of scar tissue in the spinal canal covered the dural surface in the control group. Thus, 3D-printed bionic titanium alloy artificial lamina can prevent epidural adhesion after laminectomy, while restoring the structural stability of the posterior complex, suggesting the potential of lamina substitutes for adhesion prevention after laminectomy.

Exploring the Protective Effect and Mechanism of Buddlejae Flos on Sodium Selenite-Induced Cataract in Rats by Network Pharmacology, Molecular Docking, and Experimental Validation.

Objective: Buddlejae Flos has a long history of utilization by humans to treat ophthalmic diseases. Although in vitro study revealed that it can be used for treating cataract, the bioactive components and the mechanism of efficacy remained unclear. This study aims to discover the bioactive components and mode of efficacy of Buddlejae Flos in cataract treatment. Methods: Several databases were screened for bioactive components and corresponding targets, as well as cataract-related targets. Using the String database, common targets were determined and utilized to construct protein-protein interactions (PPI). The drug-component-target-disease network map was drawn using Cytoscape software. R language was utilized to execute Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) pathway enrichment analysis. Molecular docking was done through Schrödinger Maestro software utilization. Luteolin's (LUT) effect on cataract induced by sodium selenite in rat pups was evaluated. Results: Six bioactive components with 38 common targets were identified as being associated with cataract. TP53, AKT1, EGFR, CASP3, TNF, ESR1, INS, IL6, HIF1A, and VEGFA were identified as core targets in PPI analysis, and the binding energy of LUT with AKT was the lowest. LUT has been demonstrated to significantly lower MDA levels, raise glutathione (GSH) levels, and boost the activity of antioxidant enzymes like GST, SOD, GPx, and CAT. After LUT treatment, TNF-a, IL-2, and IL-6 levels were significantly lowered. Bcl-2 mRNA expression levels and p-PI3K and p-AKT protein expression were significantly elevated. In contrast, caspase-3 and Bax mRNA expression levels were significantly decreased. Conclusion: This study demonstrates that LUT is a possible bioactive component that may be utilized for cataract treatment. Its mode of action includes oxidative stress suppression, reducing inflammation, and inhibiting apoptosis via regulating the PI3K/AKT single pathway.

[Surgical treatment for terrible triad of the elbow through medial and lateral approach].

OBJECTIVE: To investigate the clinical effects of medial and lateral approach in treating terrible triad of the elbow. METHODS: From May 2010 from May 2014, 11 patients with terrible triad of the elbow were treated through medial and lateral approach. There were 6 males and 5 females, aged from 25 to 56 years with an average of 35.2 years old. The time from injury to operation was from 1 to 13 days with an average of 5.9 days. Fracture of radial head according to Mason typing, 2 cases were type I, 7 cases were type II, 2 cases were type III. Ulnar coronoid fracture according to Regan-Morrey typing, 3 cases were type I, 7 cases were type II, 1 case was type III. Postoperative complications were observed and Mayo elbow performance score(MEPS) was used to assess the elbow joint function. RESULTS: All patients were followed up from 6 to 24 months with an average of 15.5 months. All fractures obtained healing with an average time of 14 weeks (ranged from 10 to 18 weeks). According to Mayo to assess the results, total score was 78.2±11.7, 2 cases got excellent results, 7 good, 1 fair, 1 poor. At final follow up, the mean range of motion was (108±21)° in flexion, (12±8)° in extension, (66±13)° in pronation, (28±18)° in supination. The varus angle of the elbow ranged from 5°to 8° in 3 cases and the valgus angle was 8° in 1 case. CONCLUSIONS: Treatment of the terrible triad of the elbow through medial and lateral approach can obtain satisfactory clinical effects, restore the elbow stability, allow early motion postoperatively, and promote the joint functional rehabilitation.

[Establishment of RNA interfering retrovirus vector targeting CXCR4 gene driven by human prostate-specific antigen promoter and its biological effects on prostate cancer cells].

OBJECTIVE: To construct RNA interfering retrovirus vector targeting CXCR4 gene driven by human prostate-specific antigen promoter and investigate its targeted inhibition effects in androgen-responsive prostate cancer cells LNCaP. METHODS: To clone the CXCR4 targeting siRNA gene by PCR. The PCR products were inserted into the pGensil-1 plasmid containing U6 promoter and EGFP. U6 promoter was replaced by hPSA promoter. Then, the recombinant EGFP-hPSA-siCXCR4 fragment was sub-cloned into pLXSN, which was evaluated by restriction enzyme. The pLXSN-EGFP-hPSA-siCXCR4 was transfected into PA317 cells with Lipofectamine 2000. The virus obtained from transfected PA317 cells was transfected into PC-3m, LNCaP and MCF-7 cells, respectively. The expression of CXCR4 mRNA and protein was detected by RT-PCR and Western blot. The invasion ability of prostate carcinoma cells was detected by Transwell experiment. RESULTS: The recombinant pLXSN-hPSA-siCXCR4 was successfully constructed. The expression of CXCR4 mRNA and protein in LNCaP cells was blocked by pLXSN-hPSA-siCXCR4. The expression inhibition rate was (81.53 +/- 10.22)% at mRNA level detected by semi-quantitive RT-PCR and (90.52 +/- 9.31)% at protein level detected by Western blot, respectively, in LNCaP cells at 48 h. The expression of CXCR4 mRNA and protein was effectively inhibited by sequence-specific hPSA-siCXCR4 in LNCaP cells, but not in PC-3m and MCF-7 cells. The results of Transwell experiment showed that the number of cells in down-pore of micro-membrane was 139.9 +/- 14. 2 in the treated group, significantly less in comparison with 348.4 +/- 36. 4 in the controlled group (P < 0.05). However, the number of PC-3m and MCF-7 cells in down-pore of micro-membrane was not significantly different among the control and treated groups (P > 0.05). CONCLUSION: The downstream siRNA controlled by hPSA promoter in retrovirus system can be expressed selectively in androgen-responsive prostate carcinoma cells, showing an apparent targeting character. RNAi targeted to CXCR4 driven by hPSA promoter has a potential value in gene therapy of androgen-responsive prostate cancer.

[Therapeutic effects of anterolateral decompression and fixation on thoracolumbar fractures complicated with incomplete paraplegia].

OBJECTIVE: To investigate the clinical effects of anterolateral decompression and fixation on thoracolumbar fractures complicated with incomplete paraplegia. METHODS: Thirty-six patients with thoracolumbar fractures complicated with incomplete paraplegia were treated with anterolateral decompression and fixation. RESULTS: The patients were followed up for an average of 18 months, which showed satisfactory recovery of the intervertebral space height and thoracolumbar vertebral curvature. The average Cobb's angle, spinal canal index and Frankel were improved remarkably, and none of the patients developed such complications as break or mobilization of the plate screw. CONCLUSION: Anterolateral decompression and fixation can directly and completely decompress the vertebral canal, promote the functional recovery of the spinal nerves and reconstruct the alignment of the spine as an ideal approach for treatment of thoracolumbar fractures with obvious spinal canal-occupying lesions or severe kyphos complicated with incomplete paraplegia.

[Clinical analysis of 19 elderly patients with displaced proximal humeral two- and three-part fractures].

OBJECTIVE: To analyze the surgical approaches for treating displaced proximal humeral two-and three-part fractures in elderly patients. METHODS: Nineteen elderly patients with displaced proximal humeral fractures were analyzed, including 13 patients with displaced two-part fractures and 6 with displaced three-part fractures. All the patients were treated by open reduction and fixation with humeral anatomical bone plates. RESULTS: The rate of excellent or good healing was 76.9% for two-part fractures without nonunion or humeral head necrosis, and was 66.7% for three-part fractures with a rate of humeral head necrosis of 16.7%. CONCLUSION: Displaced proximal humeral fractures in elderly patients should be managed with minimal open reduction and fixed with humeral anatomical bone plate.