Experimental Study on Rapid Hemostasis Using Peptide Hydrogels.

Uncontrolled hemorrhaging resulting from trauma, surgery, and disease-associated or drug-induced blood disorders can cause significant morbidities and mortalities in civilian and military populations. Self-assembling peptide nanofibers are particularly attractive due to their rapid and efficient hemostasis, biocompatibility, and wound-healing properties. In this study, we designed two types of 12-residue peptides by using a strong fishnet-like peptide sequence and a pro-cell adhesion sequence (Arg-Gly-Asp, RGD). The peptides are HN2-X-Ser-Phe-Cys-Phe-Lys-Phe-Glu-X-Arg-Gly-Asp-OH (where X is Pro or Tyr), which dissolve in deionized (DI) water and form stable and transparent functional hydrogels. Transmission electron microscopy and scanning electron microscopy demonstrated that the two peptides self-assemble into nanowebs and nanofibers, forming a fishnet-like and three-dimensional network structure. Circular dichroism and Fourier transform infrared spectroscopy analysis demonstrated that the self-assembled peptides mainly adopt a ß-sheet structure with ß-turn and α-helix as auxiliary assembly growth. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and SEM analysis showed that the cell survival rates were very good, delivering an obvious promotion of cell proliferation of fibroblasts and hepatocytes. Importantly, in vivo hemostasis delivered that the self-assembled peptide nanowebs and nanofibers had a good hemostatic effect on rat saphenous vein and liver bleeding, achieving 38 s faster hemostasis, which was better than commercial "Instantaneous" hemostatic powder. Accoupling the fast hemostasis and effective promotion of liver defect rapid repair, the peptide self-assembly strategy offers a clinically promising treatment option for life-threatening liver bleeding and serves as a renewed impetus for the development of peptide hydrogels as effective hemostatic agents.

[Efficacy of micro steel plate and Kirschner needle oblique and transverse internal fixation of adjacent metacarpal bone in the treatment of metacarpal diaphyseal oblique fracture].

OBJECTIVE: To compare the clinical efficacy of micro steel plate and Kirschner needle oblique and transverse internal fixation of adjacent metacarpal bone in the treatment of metacarpal diaphyseal oblique fracture. METHODS: Fifty-nine patients with metacarpal diaphyseal oblique fractures admitted between January 2018 and September 2021 were selected as the study subjects and divided into the observation group (29 cases) and the control group (30 cases) based on different internal fixation methods. The observation group was treated with Kirschner wire oblique and transverse internal fixation of adjacent metacarpal bones, while the control group was treated with micro steel plate internal fixation. Postoperative complications, operation time, incision length, fracture healing time, treatment cost, and metacarpophalangeal function were compared between the two groups. RESULTS: No incision or Kirschner wire infections occurred in the 59 patients, except for one in the observation group. No fixation loosening, rupture, or loss of fracture reduction occurred in any of the patients. The operation time and incision length in the observation group were (20.5±4.2) min and (1.6±0.2) cm, respectively, which were significantly shorter than those in the control group (30.8±5.6) min and (4.3±0.8) cm (P<0.05). The treatment cost and fracture healing time in the observation group were (3 804.5±300.8) yuan and (7.2±1.1) weeks, respectively, which were significantly lower than those in the control group (9 906.9±860.6) yuan and (9.3±1.7) weeks (P<0.05). The excellent and good rate of metacarpophalangeal joint function in the observation group was significantly higher than that in the control group at 1, 2, and 3 months after operation (P<0.05), but there was no significant difference between the two groups at 6 months after operation (P>0.05). CONCLUSION: Micro steel plate internal fixation and Kirschner wire oblique and transverse internal fixation of adjacent metacarpal bones are both viable surgical methods for treating metacarpal diaphyseal oblique fractures. However, the latter has the advantages of causing less surgical trauma, shorter operation time, better fracture healing, lower cost of fixation materials, and no need for secondary incision and removal of internal fixation.

New technique for the treatment of fresh bony mallet finger: A retrospective case series study.

Background: The bony mallet finger is a tear fracture of the extensor tendon, resulting in a flexion deformity of the finger, which affects both the function of the finger. The classical Ishiguro's method is associated with damage to the cartilage of the distal interphalangeal (DIP) joint and always lead to the joint stiffness. This paper explores a new technique to overcome the shortcomings of the classical Ishiguro's method and achieve better clinical efficacy. Methods: We examined 15 patients with bony mallet fingers, 9 males and 6 females, from February 2020 to June 2022, ranged from 23 to 58 years, including 1 case of index finger, 5 cases of middle finger, 3 cases of ring finger and 6 cases of little finger. The median course of the injury to surgery was 2 days (range, 1∼7 days). All had fresh closed injuries, according to the Wehbe and Schneider classification: 4 cases of type IA, 6 cases of type IB, 3 cases of type IIA and 2 cases of type IIB. All patients were treated surgically by the new technique. Post-operative follow-up was conducted to record the healing of the fracture, the pain of the affected finger and the function of joint movement. Results: The 15 cases were followed up after surgery. The median active range of motion was 65° (range, 55∼75°). The median extension deficit of DIP joint was 0° (range, 0∼11°). The median clinical healing time of the fracture was 6 weeks (range, 6∼10 weeks). None of the patients experienced significant pain. The patients were assessed according to the Crawford criteria at the final follow-up: 11 cases were assessed as excellent, 3 cases were assessed as good and 1 case was assessed as fair. No loss of fracture repositioning, loosening of internal fixation, skin necrosis or infection was observed. Conclusion: The application of the new technique for the treatment of bony mallet fingers has the advantages of good stability, fracture healing and functional recovery of the DIP joint, making it an ideal surgical procedure for the treatment of fresh bony mallet fingers.

Surgery combined with antibiotics for thoracic vertebral Escherichia coli infection after acupuncture: A case report.

BACKGROUND: Acupuncture is relatively popular worldwide, but an unregulated operation can easily lead to infections. The purpose of this report was to analyze a clinical case of surgery combined with the use of antibiotics for the treatment of thoracic vertebral infection by Escherichia coli (E. coli) after acupuncture. CASE SUMMARY: A 63-year-old male was diagnosed with E. coli infection in the thoracic vertebra after acupuncture. His fever and pain did not improve after treatment with broad-spectrum antibiotics for 10 d. Thus, debridement of the infected area and biopsy were decided. The final pathology confirmed the diagnosis of vertebral infection by E. coli. The patient underwent anterior and posterior thoracic vertebral debridement and internal fixation surgery combined with the use of sensitive antibiotics. He had no fever or backache 3 mo postoperatively. CONCLUSION: In this report, we first considered antibiotic treatment for the patient with septic spinal infection, but the effect was not obvious. Interventional surgery was combined with the use of sensitive antibiotics to relieve backache, and good clinical results were achieved. Furthermore, acupuncture practitioners should pay attention to hygienic measures.

A retrospective assessment of the clinical efficacy of different internal fixation methods in the treatment of distal fibula fractures in the elderly.

An ankle fracture is a fracture of the distal tibia or fibula that forms the ankle joint, usually associated with ligament and soft tissue injury, and is a common type of lower limb fracture and one of the most common types of fracture in the elderly. Although ankle fractures are one of the most common injuries seen by orthopedic trauma surgeons, there is no uniform protocol for the diagnosis and treatment of ankle fractures in the elderly, and there are many controversial indications for surgery. The aim of this study is to assess the clinical efficacy of different internal fixation methods in the treatment of distal fibular fractures in the elderly, in an effort to improve the rational selection and application of clinical acts. A retrospective analysis was performed on 68 cases of patients who suffered an ankle fracture and were treated with different internal fixation methods according to the fracture types and individual differences in distal fibula fractures. The postoperative therapeutic effect assessment was performed in terms of clinical examination, imaging evaluation, and AOFAS ankle-hind foot function scoring. There was no unhealed bone, ankle instability and loose/fractured internal fixation. Fracture healing time was 2.7 to 4.0 months (average 3.2 months). AOFAS score was 88.3 ± 6.2, of which, 34 excellent cases, 30 good cases, and 4 fair cases. Ankle activity dorsiflexion 6º~18º, average 15º; plantar flexion 26º~47º, average 37º. A good clinical efficacy could be achieved from the most appropriate individualized internal fixation for distal fibula fractures of elderly patients.

Regulation of osteoblast autophagy based on PI3K/AKT/mTOR signaling pathway study on the effect of ß-ecdysterone on fracture healing.

OBJECTIVES: To investigate the effects of ß-ecdysterone on fracture healing and the underlying mechanism. METHODS: MTT assay was used to detect the cell viability. AO/PI and flow cytometry assays were used to determine the apoptotic rate. The expression level of RunX2, ATG7 and LC3 was evaluated by qRT-PCR and Western blot assays. X-ray and HE staining were conducted on the fractured femur. Immunohistochemical assay was used to detect the expression level of Beclin-1 and immunofluorescence assay was used to measure the expression level of LC3 in the fractured femurs. Western blot was utilized to determine the expression level of PI3K, p-AKT1, AKT1, p-mTOR, mTOR, p-p70S6K, and p70S6K. RESULTS: The ALP activity and the expression of RunX2 in fractured osteoblasts were significantly elevated, the apoptotic rate was suppressed by rapamycin, 60, and 80 µM ß-ecdysterone. The state of autophagy both in fractured osteoblasts and femurs was facilitated by rapamycin and ß-ecdysterone. Compared to control, Garrett score was significantly promoted in rapamycin and ß-ecdysterone groups, accompanied by ameliorated pathological state. Lastly, the PI3K/AKT/mTOR pathway both in fractured osteoblasts and femurs was inhibited by rapamycin and ß-ecdysterone. CONCLUSION: ß-ecdysterone might facilitate fracture healing by activating autophagy through suppressing PI3K/AKT/mTOR signal pathway.

Icariin alleviates osteoarthritis by regulating autophagy of chondrocytes by mediating PI3K/AKT/mTOR signaling.

Osteoarthritis (OA) is a chronic degenerative disease that significantly impacts the quality of life of the elderly population. Recently, the pathogenesis of OA has been reported to involve autophagy in chondrocytes. Intriguingly, icariin, one of the main components of epimedium, exerts multiple pharmacological effects, including a protective effect against chondrocyte damage. Thus, we aimed to investigate the therapeutic effect of icariin on OA and its potential underlying mechanism by using a rat model of OA. After treatment with icariin or an autophagy activator (rapamycin) or inhibitor (3-methyladenine), OA chondrocyte viability was measured using the CCK-8 assay, apoptosis in the chondrocytes was evaluated using the acridine orange-propidium iodide assay and flow cytometry, and OA tissue pathological state was assessed using micro-CT scanning and safranin O staining. Furthermore, immunohistochemical staining was used to measure the expression level of Beclin-1 and immunofluorescence labeling was used to visualize LC3 expression, and western blotting was used to determine the expression levels of autophagy proteins and key proteins in the PI3K signaling pathway. The apoptotic rate of OA chondrocytes was markedly elevated by 3-methyladenine and suppressed by rapamycin and icariin; autophagy genes were drastically downregulated in the 3-methyladenine group and upregulated in the rapamycin and icariin groups; and the PI3K/AKT/mTOR signaling pathway was activated by 3-methyladenine and inhibited by rapamycin and icariin. Notably, following treatment with rapamycin and icariin, the severe pathological state in OA cartilage tissues was substantially alleviated, and this was accompanied by activated autophagy and inhibited PI3K signaling in the cartilage tissues. Taken together, these findings indicate that icariin alleviates OA by regulating the autophagy of chondrocytes by mediating PI3K/AKT/mTOR signaling.

ß-ecdysterone alleviates osteoarthritis by activating autophagy in chondrocytes through regulating PI3K/AKT/mTOR signal pathway.

PURPOSE: To investigate the therapeutic effects of ß-ecdysterone on osteoarthritis (OA) and the underlying mechanism. METHODS: OA model was established on rats by injecting MIA. ELSA was used to determine the concentration of IL-1ß, IL-6, NO and TNF-α in the chondrocytes and cartilage tissues. Immunofluorescence assay was used to determine the expression of collagen II in the chondrocytes. The survival rate of chondrocytes was evaluated by MTT assay. The apoptosis of chondrocytes was checked by AO/PI staining and flow cytometry assay. The expression level of Atg7, PI3K and caspase-3 was evaluated by qRT-PCR. Western Blot was used determine the expression of PI3K, p-AKT1, AKT1, p-mTOR, mTOR, p70S6K, p-p70S6K, LC3I, LC3II and caspase-3. HE staining was used to check the pathological state of cartilage tissues. RESULTS: Chondrocytes were tolerable to rapamycin, 3-methyladenine and ß-ecdysterone at the concentration of 10 mM, 100 nM and 40 µM, respectively. The apoptosis of chondrocytes was inhibited by rapamycin and ß-ecdysterone, and induced by 3-methyladenine. PI3K, p-AKT1, p-mTOR, p-p70S6K and caspase-3 were down-regulated by rapamycin and ß-ecdysterone, and up-regulated by 3-methyladenine in both the chondrocytes and the cartilage tissues. The expression of Atg7 and LC3II/LC3I were regulated in a opposite way. The inflammation state was improved by rapamycin and ß-ecdysterone both the chondrocytes and the cartilage tissues. HE staining results showed that the pathological state of cartilage tissues was alleviated by ß-ecdysterone. CONCLUSION: ß-ecdysterone might alleviate osteoarthritis by activating autophagy in chondrocytes through regulating PI3K/AKT/mTOR signal pathway.

Lipoxin A4 protects rat skin flaps against ischemia-reperfusion injury through inhibiting cell apoptosis and inflammatory response induced by endoplasmic reticulum stress.

BACKGROUND: The ischemia-reperfusion (I/R) injury of skin flap is a complex pathophysiological process involving many cells and factors. Although endoplasmic reticulum (ER) stress-induced cell apoptosis and inflammatory response are of immense importance in the skin flap ischemia, the treatment for I/R injury induced by ER stress is barely reported. METHODS: Healthy male Wister rats were randomly divided into three groups: sham-operated group, I/R model group and I/R + LXA4 group. I/R-induced injury in skin flaps with or without pre-treatment of Lipoxin A4 (LXA4, 100 µg/kg) was tested by using HE and TUNEL staining. Related factors associated with oxidative stress, apoptosis, inflammatory response, and ER stress were tested by ELISA, biochemical assay, and western blotting, respectively. RESULTS: Our results showed that LXA4 treatment significantly promotes skin flap survival and attenuates I/R injury by inhibiting oxidative stress, apoptosis, and inflammatory factor release, evidenced by the decreased expression of malondialdehyde (MDA), lactate dehydrogenase (LDH), NF-κBp65, tumor necrosis factor α (TNF-α), ET, active Caspase-3 and Bax and up-regulated superoxide dismutase (SOD), glutathione (GSH) level and Bcl-2 expression. Moreover, LXA4 treatment also reverses the increased expression of GRP78, p-PERK, p-eIF2α, ATF4, and CHOP induced by I/R injury. CONCLUSIONS: In conclusion, we showed that ER stress causes cell apoptosis and inflammatory response, resulting in the skin flaps injury. LXA4 exhibits a protective effect on skin flaps against I/R injury through the inhibition of ER stress.

Neo-debromoaplysiatoxin C, with new structural rearrangement, derived from debromoaplysiatoxin.

Neo-debromoaplysiatoxin C (1), a new member of the aplysiatoxin family, was isolated from the marine cyanobacterium Lyngbya sp. The structure of 1 was elucidated based on spectroscopic data, and its stereochemistry was determined from NOESY spectrum and biosynthetic considerations. This new compound presents an intriguing 10-membered lactone ring skeleton derived from debromoaplysiatoxin by structural rearrangement, which is the first example in the aplysiatoxin family. Its biological properties were evaluated for cytotoxicity, PKCδ activation and inhibitory effects on potassium channel.

Two New Neo-debromoaplysiatoxins-A Pair of Stereoisomers Exhibiting Potent Kv1.5 Ion Channel Inhibition Activities.

A pair of stereoisomers possessing novel structures with 6/6/5 fused-ring systems, neo-debromoaplysiatoxin E (1) and neo-debromoaplysiatoxin F (2), were isolated from the marine cyanobacterium Lyngbya sp. Their structures were elucidated using various spectroscopic techniques including high resolution electrospray ionization mass spectroscopy (HRESIMS) and nuclear magnetic resonance (NMR). The absolute stereochemistry was determined by calculated electronic circular dichroism (ECD) and gauge-independent atomic orbital (GIAO) NMR shift calculation followed by DP4+ analysis. Significantly, this is the first report on aplysiatoxin derivatives with different absolute configurations at C9-C12 (1: 9S, 10R, 11S, 12S; 2: 9R, 10S, 11R, 12R). Compounds 1 and 2 exhibited potent blocking activities against Kv1.5 with IC50 values of 1.22 ± 0.22 µM and 2.85 ± 0.29 µM, respectively.

The facile synthesis of copper oxide quantum dots on chitosan with assistance of phyto-angelica for enhancing the human osteoblast activity to the application of osteoporosis.

Osteoblasts are an important key factor for the pathogenesis of several bone-related diseases, notably in osteoporosis. Osteoporosis is a disorder categorized based on the bone mineral density (BMD) and an alteration in the bone micro-architecture had been considered as the major determinant for increasing the fracture risk. The available medicine for curing the osteoporosis shows a minimal or no activity against the genesis or function of osteoblasts. The present study was conducted to determine the influence of phyto Angelica species (Ang.) mediated synthesized copper quantum dots decorated chitosan on human osteoblast cells for application of osteoporosis. The phyto compound of Angelica sp. was extracted by ethanol as solvent and it has been characterized through spectral analyses. An Angelica sp. mediated synthesized copper oxide quantum dots (CuO QDs) and the presence of CuO QDs on chitosan have been analyzed and exhibited by important spectral investigations. The morphological observation of CuO QDs on the chitosan (CS) was visualized by the microscopic analyses. The MTT assay results showed that cell growth of CuO QDs/CS-Ang. by the concentration dependent. The highest cell growth (87%) was noted at 5 µg/mL followed by 80 and 77% at 15 and 25 µg/mL respectively. The functional groups and potential compounds of Angelica sp. with CuO QDs/CS has been improved the osteoblast cell activity as prophylactic potentials against osteoporosis.

Chemical and biological study of aplysiatoxin derivatives showing inhibition of potassium channel Kv1.5.

Three new aplysiatoxins, neo-debromoaplysiatoxin D (1), oscillatoxin E (2) and oscillatoxin F (3), accompanied by four known analogues (4-7), were identified from the marine cyanobacterium Lyngbya sp. Structural frames differ amongst these metabolites, and therefore we classified compounds 1 and 4-6 as aplysiatoxins as they possess 6/12/6 and 6/10/6 tricyclic ring systems featuring a macrolactone ring, and compounds 2, 3 and 7 as oscillatoxins that feature a hexane-tetrahydropyran in a spirobicyclic system. Bioactivity experiments showed that compounds 1 and 4-6 presented significant expression of phosphor-PKCδ whereas compounds 2, 5 and 7 showed the most potent blocking activity against potassium channel Kv1.5 with IC50 values of 0.79 ± 0.032 µM, 1.28 ± 0.080 µM and 1.47 ± 0.138 µM, respectively. Molecular docking analysis supplementing the binding interaction of oscillatoxin E (2) and oscillatoxin F (3) with Kv1.5 showed oscillatoxin E (2) with a strong binding affinity of -37.645 kcal mol-1 and oscillatoxin F (3) with a weaker affinity of -32.217 kcal mol-1, further supporting the experimental data.