Heat-induced antigen retrieval utilizing modified Tris-EDTA buffer for reprobing of Western blots on nitrocellulose paper.

The development of heat-induced antigen retrieval technologies with Tris-EDTA buffer has dramatically improved immunostaining of specific antigens for routine immunohistochemical detection (Krenacs et al., 2010) [1]. However, little evidence exists on whether heat-Induced antigen retrieval utilizing Tris-EDTA buffer can strip western blot (WB) membranes and allow sequential reprobing. Here, we serendipitously discover that ∼95 °C Tris-EDTA buffer with 0.01% Tween 20 could repeatedly strip the Nitrocellulose membranes (NC). After electroblotting, NC blots were soaked into Tris-EDTA stripping buffer (∼95 °C, 10-25min) and we could perform at least five rounds (the following antibodies used: Vinculin, Atg7, Caspase-3, UBA5, JNK and ERK1/2) stripping in sequential chemiluminescent detections. The NC membranes also show clear western signals and background without losing transferred proteins during the reprobing process of WB. Hence, this study report additional new roles of the heat-Induced antigen retrieval Tris-EDTA buffer with 0.01% Tween 20. The method is simpler, more affordable and harmless for the nitrocellulose paper, which will be helpful for effective reprobing in western blotting applications.

A meta-analysis comparing the effects of cemented and uncemented prostheses on wound infection and pain in patients with femoral neck fractures.

To providing evidence-based recommendations for surgery in patients with femoral neck fractures, a meta-analysis was conducted to comprehensively evaluate the effects of cemented and uncemented prostheses on postoperative surgical site wound infection and pain in these patients. Relevant studies on the use of cemented prostheses in femoral neck fractures were retrieved from PubMed, EMBASE, Cochrane Library, Ovid, CNKI, and Wanfang databases from the time of their establishment until March 2023. Two authors independently screened and extracted data from the included and excluded literature according to predetermined criteria. Review Manager 5.4 software was used to perform meta-analyses on the collected data. A total of 27 articles comprising 34 210 patients (24 646 cases in the cemented group and 9564 cases in the uncemented group) were included in the final analysis. The results of the meta-analysis showed that, compared with the uncemented group, cemented prostheses significantly reduced the incidence of surgical site wound infections (odds ratio [OR]: 0.75, 95% confidence interval [CI]: 0.64-0.88, p < 0.001) and relieved surgical site wound pain (standardised mean difference: -0.76, 95% CI: -1.12-0.40, p < 0.001), but did not reduce the incidence of pressure ulcers after surgery (OR: 0.50, 95% CI: 0.20-1.26, p = 0.140). Therefore, existing evidence suggests that the use of cemented prostheses in femoral neck fracture surgery can significantly reduce the incidence of surgical site wound infections and relieve surgical site wound pain, which is worthy of clinical recommendation.

A Nanoparticle Platform To Evaluate Bioconjugation and Receptor-Mediated Cell Uptake Using Cross-Linked Polyion Complex Micelles Bearing Antibody Fragments.

Targeted nanomedicines are a promising technology for treatment of disease; however, preparation and characterization of well-defined protein-nanoparticle systems remain challenging. Here, we describe a platform technology to prepare antibody binding fragment (Fab)-bearing nanoparticles and an accompanying real-time cell-based assay to determine their cellular uptake compared to monoclonal antibodies (mAbs) and Fabs. The nanoparticle platform was composed of core-cross-linked polyion complex (PIC) micelles prepared from azide-functionalized PEG-b-poly(amino acids), that is, azido-PEG-b-poly(l-lysine) [N3-PEG-b-PLL] and azido-PEG-b-poly(aspartic acid) [N3-PEG-b-PAsp]. These PIC micelles were 30 nm in size and contained approximately 10 polymers per construct. Fabs were derived from an antibody binding the EphA2 receptor expressed on cancer cells and further engineered to contain a reactive cysteine for site-specific attachment and a cleavable His tag for purification from cell culture expression systems. Azide-functionalized micelles and thiol-containing Fab were linked using a heterobifunctional cross-linker (FPM-PEG4-DBCO) that contained a fluorophenyl-maleimide for stable conjugation to Fabs thiols and a strained alkyne (DBCO) group for coupling to micelle azide groups. Analysis of Fab-PIC micelle conjugates by fluorescence correlation spectroscopy, size exclusion chromatography, and UV-vis absorbance determined that each nanoparticle contained 2-3 Fabs. Evaluation of cellular uptake in receptor positive cancer cells by real-time fluorescence microscopy revealed that targeted Fab-PIC micelles achieved higher cell uptake than mAbs and Fabs, demonstrating the utility of this approach to identify targeted nanoparticle constructs with unique cellular internalization properties.

An advanced optic-fiber differential sensing system enhanced by molecularly imprinted polymer for specific sodium benzoate detection.

A molecularly imprinted polymer (MIP) based microfiber differential demodulation sensing system for sodium benzoate (SB) concentration detection is proposed. The specific binding of MIP on the surface of microfibers with SB can lead to changes in local refractive index (RI). RI change induces a drift in the interference wavelength, which can be monitored by the power difference between two fiber Bragg gratings (FBGs). The sensing system can detect SB in the concentration range of 0.1-50 µg/ml, and interference wavelength and FBG power difference sensitivities are 0.55 nm/(µg/ml) and 2.64 dB/(µg/ml) in the low concentration range of 0.1-1 µg/ml, respectively, with a limit of detection (LOD) of 0.1 µg/ml. This microfiber differential demodulation sensing system is not only simple to fabricate, but also simplifies the demodulation equipment to reduce the cost, which providing a simple, reliable and low-cost technique for the quantitative detection of SB concentration in beverages and flavoured foods.

Spatial distribution, source identification, and transportation paths of plutonium in the Beibu Gulf, South China Sea.

To investigate the spatial distribution and source of plutonium isotopes in the Beibu Gulf, surface sediments were collected and analyzed using sector field inductively coupled plasma mass spectrometry (SF-ICP-MS). The activities of 239+240Pu in surface sediments ranged from 0.012 to 0.451 mBq/g (mean: 0.171 ± 0.138 mBq/g, n = 36), indicating a decreasing trend in a counterclockwise direction from the southern bay mouth. The counterclockwise decreasing trend in the south of the bay mouth is similar to the current in the Beibu Gulf. The 240Pu/239Pu atom ratios in surface sediments ranged from 0.156 to 0.283 (mean: 0.236 ± 0.031, n = 36), slightly higher than that of the global fallout value of 0.18. This suggests that the Pu in the Beibu Gulf was a combination of global fallout and Pacific Proving Ground (PPG). The average contribution of the plutonium (Pu) derived from the PPG in the sediment was estimated to be 52 % ± 24 %.

Treating Osteoporotic Spinal Fractures in Elderly Patients via a Modified Clinical Bone Cement Injection Technique.

The minimally invasive injection of bone cement (MIIBC) is an effective way to treat senile osteoporotic spinal fractures (OSF) in clinical practice. However, the intraspinal dura and nerves may be damaged when the puncture needle passes through the pedicle. Therefore, in this protocol, the puncture site was optimized during the surgery, selecting the same 1-2 cm away from the surface projection of the diseased vertebra. The needle was punctured along the lateral cortex of the pedicle from the junction of the pedicle and the vertebral body into the vertebral body. Meanwhile, bone cement was used as a filling material, and the MIIBC was performed by a percutaneous puncture at the external edge of the pedicle under C-arm fluoroscopy. This modified puncture site is far away from the spinal canal as possible, thereby reducing the risk of the puncture needles penetrating the spinal canal and damaging the nerves and dura mater. In conclusion, a modified MIIBC by percutaneous lateral pedicle puncture can effectively relieve pain in elderly patients with OSF.

Hydrogel Complex Electrospun Scaffolds and Their Multiple Functions in In Situ Vascular Tissue Engineering.

Hydrogel complex scaffolds (hydrogel scaffolds) are prepared by coating precursor solutions onto heparin-modified poly(ε-caprolactone) (PCLH) scaffolds followed by subsequent in situ gelation. Here, we show that hydrogel complexation can significantly strengthen the scaffold and slow its degradation. The hydrogel scaffold was implanted into the abdominal aorta of a rat model, and the aneurysm incidence rate of the hydrogel scaffolds sharply decreased compared with that of the hydrogel-free scaffolds. Histological and immunohistological analyses showed that the implanted grafts had good vascular regeneration. The absence of calcification and occurrence of contractile smooth muscle cells (SMCs) at the first month was found in the hydrogel-free PCLH scaffold due to the presence of surface-modified heparin, whereas the hydrogel scaffold exhibited mild calcification and later occurrence of contractile SMCs as the complexed hydrogel covered the fibers and blocked the interaction between heparin and cells. Heparin was further physically encapsulated into the hydrogel before gelation, and its sustainable release was demonstrated by an in vitro release test. A pilot implantation in a rabbit carotid model showed that the encapsulated heparin modulated the scaffold characteristics including anticoagulation, anticalcification, and the early occurrence of contractile SMCs in vivo. Consequently, hydrogel complexation can significantly improve the in vivo regeneration property of the scaffold due to its multiple beneficial characteristics.

[The effect of orthognathic surgery on speech function in patients with skeletal Class â ¢ malocclusion].

PURPOSE: To investigate the influence of the position of the upper and lower jaws on the anatomical structure of pharynx before and after orthognathic surgery in patients with skeletal Class Ⅲ malocclusion. METHODS: Craniofacial CT scan and speech data were collected from 31 patients with skeletal Class Ⅲ malocclusion before and 3 months after surgery. The collected CT data was imported into Dolphin imaging 11.95 software to establish a digital original model, and the anatomical structure of the pharynx was measured and analyzed. Speech data were analyzed objectively and subjectively by Computerized Speech Lab 4500b and professional speech specialists. Statistical analysis was performed using SPSS 24.0 software package. RESULTS: The distance from the lower edge of the soft palate to the posterior pharyngeal wall, the shortest distance from the posterior margin of the tongue to the posterior pharyngeal wall and its corresponding cross-sectional area were significantly different from those before surgery (P<0.05). The changes of SNA, SNB, ANB, OJ, and OBJ before and after surgery were significant in this series. Importantly, the speech intelligibility of orthognathic patients before and after surgery changed significantly subjectively (P<0.05). Objectively, the postoperative vowels /a/B2, B3, B4, /i/B1,B2, /u/B1,B2 and B4 of the patients were significantly different from those before surgery. There was no significant difference in the lower limit frequency of the consonants /x/, /zh/, /s/, the energy value of /zh/ and the grammatical form of /z/ before and after surgery. The maxillary advancement distance was highly correlated or significantly correlated with △S1, △VOP, and voice changes. CONCLUSIONS: Orthognathic surgery moves the upper and lower jaws to cause changes in the anatomy of the pharyngeal cavity, leading to changes of postoperative speech.