Ce6-modified Fe ions-doped carbon dots as multifunctional nanoplatform for ferroptosis and photodynamic synergistic therapy of melanoma.

BACKGROUND: Despite the higher sensitivity of melanoma towards ferroptosis and photodynamic therapy (PDT), the lack of efficient ferroptosis inducers and the poor solubility of photosensitizers restrict their synergistic strategies. With unique advantages, carbon dots (CDs) are expected to serve as innovative building blocks for combination therapy of cancers. RESULTS: Herein, an ferroptosis/PDT integrated nanoplatform for melanoma therapy is constructed based on chlorin e6-modified Fe ions-doped carbon dots (Fe-CDs@Ce6). As a novel type of iron-carbon hybrid nanoparticles, the as-prepared Fe-CDs can selectively activate ferroptosis, prevent angiogenesis and inhibit the migration of mouse skin melanoma cells (B16), but have no toxicity to normal cells. The nano-conjugated structures facilitate not only the aqueous dispersibility of Ce6, but also the self-accumulation ability of Fe-CDs@Ce6 within melanoma area without requiring extra targets. Moreover, the therapeutic effects of Fe-CDs@Ce6 are synergistically enhanced due to the increased GSH depletion by PDT and the elevated singlet oxygen (1O2) production efficiency by Fe-CDs. When combined with laser irradiation, the tumor growth can be significantly suppressed by Fe-CDs@Ce6 through cyclic administration. The T2-weighted magnetic resonance imaging (MRI) capability of Fe-CDs@Ce6 also reveals their potentials for cancer diagnosis and navigation therapy. CONCLUSIONS: Our findings indicate the multifunctionality of Fe-CDs@Ce6 in effectively combining ferroptosis/PDT therapy, tumor targeting and MRI imaging, which enables Fe-CDs@Ce6 to become promising biocompatible nanoplatform for the treatment of melanoma.

Molecular Etiology in Thumb Polydactyly in Mutated GLI3 Mouse Gene.

Sporadic thumb polydactyly with nonfamily inheritance is the most common in clinical work. This study focused on characterization of GLI3 gene function. We constructed the plasmid with p.m948i point mutation of GLI3 and transfected it into mouse embryonic fibroblasts (MEFs) to study the effects and potential mechanism of the mutant gene. The RNA of GLI3 mutant cells was extracted and analyzed by transcriptome sequencing and bioinformatics. Finally, we constructed cbx3 overexpression plasmid, designed siRNA for gene silencing, and transfected it into the MEFs. Cell proliferation and invasion ability of the MEFs were examined. The results showed that there were 2,452 differential expression genes in the MEFs transfected with GLI3 mutant plasmid compared with wild-type MEFs. The results of differential expression analysis showed that the cbx3 gene was significantly up-regulated. Overexpression of cbx3 in MEFs promoted cell proliferation and invasion, while siRNA knockdown of cbx3 expression reduced proliferation and invasion. GLI3 gene mutation in MEFs resulted in cbx3 up-regulation and promoted MEF proliferation and invasion. This study further clarified the potential function of GLI3 in limb development, established a new relationship between gene mutation and polydactyly, and preliminarily clarified the possible signal pathway, all of which have laid a foundation for further study on the etiology of polydactyl.

Efficacy of Platelet-rich Plasma for Low Back Pain: A Systematic Review and Meta-analysis.

BACKGROUND: Platelet-rich plasma (PRP) may be beneficial for patients with low back pain. However, the results remain controversial. We conducted a systematic review and meta-analysis to explore the efficacy of PRP for low back pain. METHODS: PubMed, Embase, Web of Science, EBSCO, and Cochrane Library databases were searched systematically. Randomized controlled trials (RCTs) assessing the effect of PRP on low back pain were included. Two investigators independently searched articles, extracted data, and assessed the quality of included studies. The primary outcome was pain scores within 8 weeks. Meta-analysis was performed using the random-effects model. RESULTS: Three RCTs involving 131 patients were included in the meta-analysis. Overall, compared with control intervention for low back pain, PRP injection was found to reduce pain scores significantly (mean difference: - 1.47; 95% confidence interval [CI], - 2.12 to - 0.81; p < 0.0001), improve the number of patients with > 50% pain relief at 3 months (risk ratio [RR]: 4.14; 95% CI, 2.22-7.74; p < 0.00001), and offer relatively good patient satisfaction (RR: 1.91; 95% CI, 1.04-3.53; p = 0.04). No increase in adverse events was reported after PRP injection (RR: 1.92; 95% CI, 0.94-3.91; p = 0.07). CONCLUSIONS: Compared with control intervention for low back pain, PRP injection was found to improve pain relief and patient satisfaction significantly with no increase in adverse events.

Identification of novel mutations in preaxial polydactyly patients through whole-exome sequencing.

BACKGROUND: Polydactyly is one of the most common hereditary limb malformation characterized by additional digits in hands and/or feet. With extra fingers/toes, which could be very problematic, polydactyly patients are usually treated in early childhood by removing of extra digits with surgery. Genetically, polydactyly is caused by mutations of genes that involve in digit formation. METHODS: In the current report, we performed genetic analysis for polydactyly using DNA samples from a cohort of 20 Chinese patients. All patients show preaxial polydactyly in one of their hands. RESULTS: With whole-exome sequencing (WES), we have identified two novel heterozygous mutations c.G2844A in GLI3 gene (OMIM 165240) and c.1409_1410del in EVC gene (OMIM 604831). Compound heterozygous mutations that affect KIAA0586 gene (OMIM 610178) are also detected. Proteins encoded by the genes have important roles in primary cilia and regulate sonic hedgehog signaling pathway. CONCLUSION: Our study highlights the important roles of primary cilia in limb development, and helps to further understand the molecular mechanisms for polydactyly formation.

Alterations of signaling pathways in muscle tissues of patients with amyotrophic lateral sclerosis.

INTRODUCTION: Amyotrophic lateral sclerosis (ALS), a degenerative disorder of the central nervous system, manifests as progressive weakening of muscles. The diagnosis and prognosis of ALS are often unclear, so useful biomarkers are needed. METHODS: Total proteins were extracted from muscle samples from 36 ALS, 17 spinal muscular atrophy (SMA), and 36 normal individuals. The expression levels of 134 proteins and phosphoproteins were assessed using protein pathway array analysis. RESULTS: Seventeen proteins were differentially expressed between ALS and normal muscle, and 9 proteins were differentially expressed between ALS and SMA muscle. The low-level expression of Akt and Factor XIIIB correlates with unfavorable survival, and the risk score calculated based on these proteins predicts the survival of each individual patient. CONCLUSIONS: Some proteins could be selected as clinically useful biomarkers. Specifically, Akt and Factor XIIIB were found to be promising biomarkers for estimating prognosis in ALS.

A crucial role of IL-17 and IFN-γ during acute rejection of peripheral nerve xenotransplantation in mice.

Nerve injuries causing segmental loss require nerve grafting. However, autografts and allografts have limitations for clinical use. Peripheral nerve xenotransplantation has become an area of great interest in clinical surgery research as an alternative graft strategy. However, xenotransplant rejection is severe with cellular immunity, and Th1 cells play an important role in the process. To better understand the process of rejection, we used peripheral nerve xenografts from rats to mice and found that mononuclear cells expressing IFN-γ and IL-17 infiltrated around the grafts, and IFN-γ and IL-17 producing CD4+ and CD8+ T cells increased during the process of acute rejection. The changes of IL-4 level had no significant difference between xenotransplanted group and sham control group. The rejection of xenograft was significantly prevented after the treatment of IL-17 and IFN-γ neutralizing antibodies. These data suggest that Th17 cells contribute to the acute rejection process of peripheral nerve xenotransplant in addition to Th1 cells.

[Experimental study on the functional reserve of ulnar nerve in rats].

OBJECTIVE: To study the functional change of nerve trunk after removing the partial bundles of ulnar nerve, to propose the concept of functional reserve of peripheral nerves and to investigate the functional reserve quantity of peripheral nerves. METHODS: Two hundred and twenty SD rats (male or female), aging 3 months and weighing 300-350 g, were randomized into the experimental group and the control group (n=110 per group). And the experimental group was subdivided into group 1/8, group 1/4, group 1/3, group 1/2 and group 2/3 according to the resection portion (n=22 per group). In the experimental group, the section of the lowest level on ulnar nerve trunks was exposed, and a certain portion of its bundles was separated and cut, while in the control group the bundles were only separated without resection. The general condition of all rats was observed, and the motoneurons in cornu anterius medullae spinalis were detected at 1 week, 2 weeks and 2 months after operation. The neuro-electrophysiology and the function of dominated muscles were detected at 2 weeks, 2 months, 3 months, and 4 months after operation. RESULTS: All the rats survived without infection and obvious ulcer in the limbs. The number of motoneurons in cornu anterius medullae spinalis in various experimental subgroups witnessed no obvious changes (P > 0.05). The superstructure changed obviously at the early postoperative stage in group 1/2 and group 2/3, but restored well at 2 months after operation. For the latent period of evoked potential, there was no significant difference between the various experimental subgroups and the control group at each time point (P > 0.05), but there was a significant difference among the various experimental subgroups when compared the time points of 2, 3 and 4 months to that of 2 weeks (P < 0.05) and no statistically significant difference at other time points (P > 0.05). For the wave amplitude of evoked potential of motor nerves, the maximum wave amplitude and the persistence time of the dominate muscle, there were significant differences between the various experimental subgroups and the control group at each time point (P < 0.05), and there were significant differences among the various experimental subgroups when comparing the time points of 2, 3 and 4 months to that of 2 weeks (P < 0.05) and no statistical significance at other time points (P > 0.05). CONCLUSION: The functional reserve of the ulnar nerve without compromise accounts the 1/3 of the whole trunk diameter.

[Spinal pathologic changes and axonal regeneration after brachial plexus root anastomosis in rats].

OBJECTIVE: To evaluate the feasibility of direct anastomosis in the rat model of the brachial plexus extravertebral foramen nerve root division of C5-7. Methods Forty-eight SD rats (male or female) aging 4-6 months and weighing 250-300 g were selected to make the model of extravertebral foramen nerve root division of C5-7. The left C5-7 nerve roots, as the experimental sides, were separated to the brachial plexus nerve trunk and the transected roots were sutured to the proximal stump immediately after cutting off the brachial plexus extravertebral foramen nerve root division. The right C5-7 nerve roots, as the control sides, received no operation. The general condition of rats after operation was observed. The gross observation, the histological observation and BDA nerve tracing technology were adopted to observe the wet weight of musculus biceps brachii, the cross section of biceps brachii and the spinal cord and distal nerve trunk at 3 weeks, 3 months and 6 months after operation. Results All rats survived well after operation. Claudication and unfold claw reflex were observed in the experimental sides, and the unfold claw reflex disappeared 3 months later. Comparatively, the control sides were normal. Nerve adhesion aggravated gradually and the neural stems were shriveled within 6 months after operation in the experimental sides. Comparatively, the control sides were normal. The wet weight of biceps brachii in the experimental sides and the control sides at 3 weeks, 3 months and 6 months after operation was (0.28 +/- 0.12), (1.37 +/- 0.33), (0.58 +/- 0.10), (1.36 +/- 0.35), (1.39 +/- 0.31), (1.37 +/- 0.38) g, respectively, indicating significant differences between two sides at 3 weeks and 3 months (P < 0.05), but no significant difference at 6 months (P > 0.05). The modified Marsland and the LFB staining of spinal cord and superior trunk of brachial plexus showed that the number of neurons, cell nuclear and Nissl body decreased and cell bodies changed from swelling to shrinkage, dyeing nerve fibers increased, neural axone was thin and myelin sheath was slightly stained at each time point in experimental side. The number of motor neurons in cornu anterius medullae spinalis in the experimental side was 84.5% +/- 3.2%, 74.4% +/- 4.5%, 73.7% +/- 3.8% of that in the control side at each time point, respectively. HE staining of biceps brachii detected that the muscle denaturation was very serious at 3 months after operation and then recovered. Neural tracing used BDA showed that the closer to the proximal of nerve trunk, the more obviously stained it was of myelin sheath and the more massive of axon at 6 months after operation. And there was almost no myelin and axon stained in musculocutaneous nerve. CONCLUSION: In the rat model of brachial plexus extravertebral foramen nerve root division, the motoneuron in cornu anterius medullae spinalis necrosis rate reaches 20%-30%, and most of the residual neurons are pathologic. The regenerated fibers manifest as insufficient dynamic power and incomplete development, making no sense for the recovery of end organ function. Therefore, the exact mechanism of the recovery of biceps brachial muscle demands further study.

[Effect of velvet antler polypeptide on peripheral nerve regeneration].

OBJECTIVE: To investigate the velvet antler polypeptide (VAP) on sciatic nerve regeneration in rats through local administration and VAP-PLGA compound membrane. METHODS: The 3, 15 mg/g of VAP-PLGA compound membrane were prepared by compounding VAP and PLGA, respectively. Seventy-two Wistar rats, male or female, aged 3-6 months and weighing (250 +/- 50) g, were selected to make the model of sciatic nerve section. Then, all rats were randomized into 4 groups (n = 18): group A in which nothing was given after anastomosis, group B in which 1 mL of VAP at the concentration of 10 mg/L was injected into the gastrocnemius muscle medial for every other day, group C in which 3 mg/g of VAP-PLGA compound membrane was given to the nerve anastomotic stoma and group D in which 15 mg/g of VAP-PLGA compound membrane was given to the nerve anastomotic stoma. The sciatic adhesion degree observation, electrophysiological examination, immunohistochemical staining and hemi-quantity calculation and horseradish peroxidase (HRP) retrograde tracing were conducted 2, 4 and 6 weeks after operation, respectively. RESULTS: All rats survived to the end of the experiment, without foot ulcer or neuroma. Severer nervous adherence was observed in group A, mild adherence in group B, and no adherence in groups C and D 2, 4 and 6 weeks after operation, respectively. The recovery rate of the evoked potential of triceps surae in groups B, C and D was better than that in group A (P < 0.01), group D was superior to groups B and C (P < 0.05) at each time point. No significant difference between group B and group C (P > 0.05) 2, 4 weeks after operation was detected, but group C was superior to group B (P < 0.05) 6 weeks after operation. For the regenerative fiber axon and the expression of myelin sheath TGF-P1 and IGF antigen, the staining intensity in groups B, C and D was higher than that in group A at each time point (P < 0.05), and there were significant differences between group D and groups B and C 6 weeks after operation (P < 0.05), but no difference between groups B and C (P > 0.05). The HRP retrograde tracing showed that the myelinated nerve fiber stained by HRP gradually increased as time passed by and myelinated nerve fiber stained by HRP in groups B, C and D was much more than that in group A, and group D was superior to the other groups. No significant difference between group B and group C was detected. CONCLUSION: To apply VAP through either local administration or VAP-PLGA compound membrane around the attached site of nerve anastomosis is capable of promoting nerve regeneration, which has an obvious dose-effect relationship with the dose of VAP. Meanwhile, VAP-PLGA compound membrane can prevent the nerve adhesion.

[Experimental study on brachial plexus injury induced by radiation in rats].

OBJECTIVE: To explore the changes of morphology and ventricornual motor neurons in SD rats' ventral horn of spinal cord after radiated as the therapy protocol for breast cancer, to discover the rule of radiation-induced injury of brachial plexus, and also if there exits the reversible conversion in neurons. METHODS: Twenty SD rats were selected. The left side of the rats was used as the radiation side, and the right side as the control side. The RIBPI animal models were established by divided-dose of radiation. Using 2 Gy/time and 5 times/week, a total administered dose reached 30 Gy after 3 weeks. The behaviour of the rats was observed after radiation. At 3, 5, 7 and 9 weeks after the last radiation (n = 4), the wet weights of biceps brachii muscle, upper-limb circumference and compound action potential were examined; the pathological changes of biceps brachii muscle, the morphological changes, counts of the motor neurons in ventral horn and axons of bilateral spinal cord were observed by HE staining, argentums staining and toluidine blue staining. RESULTS: The rats showed lameness and a "claw hand" 3 weeks after radiation. Compared with control side, the wet weights of biceps brachii muscle and upper-limb circumference were significantly reduced, meanwhile, the compound action potential significantly decreased, and its latent period was also significantly prolonged 3, 5, 7 and 9 weeks (P < 0.05). The histological observation: Musculocutaneous nerve showed decreased medullated fibers, heterogeneous ditribution and decreased density, thin myelin sheath, damaged nerve structure and collagen hyperplasia; biceps brachii muscle showed degeneration, fiber breakage and inflammatory cell infiltration; The account of motor neurons in ventral horn was significantly decreased in the radiation side with time extending, the sign of cell death, such as, the neurons crimple, and karyolysis were observed (P < 0.05). CONCLUSION: Large dose of X-ray can induced brachial plexus injury, and the lameness, a "claw hand", biceps brachii muscle atrophy and the compound action potential abnormality. The account of motor neurons in ventral horn was significantly decreased. The motor neurons showed oxonal degeneration and myelinec degeration.