Gelatin-based biomaterials and gelatin as an additive for chronic wound repair.

Disturbing or disrupting the regular healing process of a skin wound may result in its progression to a chronic state. Chronic wounds often lead to increased infection because of their long healing time, malnutrition, and insufficient oxygen flow, subsequently affecting wound progression. Gelatin-the main structure of natural collagen-is widely used in biomedical fields because of its low cost, wide availability, biocompatibility, and degradability. However, gelatin may exhibit diverse tailored physical properties and poor antibacterial activity. Research on gelatin-based biomaterials has identified the challenges of improving gelatin's poor antibacterial properties and low mechanical properties. In chronic wounds, gelatin-based biomaterials can promote wound hemostasis, enhance peri-wound antibacterial and anti-inflammatory properties, and promote vascular and epithelial cell regeneration. In this article, we first introduce the natural process of wound healing. Second, we present the role of gelatin-based biomaterials and gelatin as an additive in wound healing. Finally, we present the future implications of gelatin-based biomaterials.

miR-708-3p promotes gastric cancer progression through downregulating ETNK1.

MicroRNAs (miRNAs) are small, evolutionarily conserved, non-coding RNAs playing a role in the proliferation, metastasis, apoptosis, chemo-sensitivity, and chemo-resistance of gastric cancer, as well as the stemness of gastric cancer stem cells. miR-708-3p induces gastric cancer cell chemo-resistance, but its actual role in gastric cancer progression remains unclear. This paper shows that miR-708-3p is upregulated in gastric cancer samples and that a high miR-708-3p expression in gastric cancer patients is associated with poor overall survival. Our functional study results indicate that miR-708-3p overexpression promotes gastric cancer cell proliferation and migration, inhibits cell apoptosis, and facilitates the transition from the G0/G1 to the G2/M phase. Furthermore, reducing miR-708-3p levels yielded opposite effects. Next, our in vivo experiments revealed that miR-708-3p advanced gastric cancer cell growth in nude mice. The underlying mechanism was the regulation of ethanolamine kinase 1 (ETNK1) expression by miR-708-3p, which bound to the 3'UTR of the ETNK1 gene in gastric cancer cells. Finally, the recovery assay results showed that ETNK1 overexpression could slow miR-708-3p-induced gastric cancer progression. In conclusion, we identified a new miR-708-3p/ETNK1 pathway involved in gastric cancer progression. These results may offer new targets for gastric cancer therapy and markers for gastric cancer prognosis.

[Methodological Evaluation of Microarray in the Detection of α-Thalassemia].

OBJECTIVE: To proceed the clinical evaluation of DNA microarray for thalassemia gene detection. METHODS: Peripheral blood samples of 166 thalassemia gene test subjects were collected and tested for thalassemia genes by microarray chip method and Gap-PCR method combined with PCR-reverse dot blot hybridization method according to double-blind control test. The specificity, sensitivity, positive predictive value, negative predictive value, and total coincidence rate of the microarray chip method were evaluated. When the two methods were inconsistent, multiplex ligation dependent probe amplification (MLPA) was used to verify the deletional α-thalassemia. RESULTS: Compared with Gap-PCR method, specificity, sensitivity, positive predictive value, negative predictive value, Youden index, and total coincidence rate of microarray chip method was 100% (70/70), 96.88% (93/96), 100% (93/93), 95.89% (70/73), 0.969, and 97.59% (162/166), respectively, while compared with PCR-reverse dot blot hybridization method was 100% (125/125), 100% (41/41), 100% (41/41), 100% (125/125), 1, and 100% (166/166), respectively. CONCLUSION: The microarray chip method for α-thalassemia gene detection shows the advantages of high specificity, sensitivity, and throughput.

[Application of DNA Microarray in Genetic Mutation Detection in Patients with Thalassemia].

OBJECTIVE: To perform dried blood spots thalassemia gene detection in patients with positive blood phenotypes by microarray technology, and evaluate its value in clinical detection. METHODS: DNA samples were extracted from dried blood spots of 410 patients. Microarray technology was used to detect 3 deletion and 3 non-deletion types of α-thalassemia and 19 ß-thalassemia point mutations which were common gene mutions in China. RESULTS: There were 357 positive cases in all the 410 tested samples with the positive rate 87.07%, among which 299 cases (72.93%) carried deletion or point mutations of α-thalassemia, 29 cases (7.07%) carried point mutations of ß-thalassemia and 29 cases (7.07%) carried gene mutations of complex αß-thalassemia syndrome. The mutations of α-thalassemia were involved with --SEA heterozygous deletion (177 cases, 59.2%), αCS heterozygote (60 cases, 20.07%) and several other genotypes. The common mutations of ß- thalassemia were involved with ßCD41-42 heterozygote (10 cases, 34.48%) and ßCD17 heterozygote (9 cases, 31.03%). The mutations of complex αß-thalassemia syndrome were mainly involved with --SEA/αα+ßCD17/ßN (7 cases, 24.14%), αCSα/αα + ßCD41-42/ßN (3 cases, 10.34%) and -α4.2/αα + ßCD17/ßN (3 cases, 10.34%). CONCLUSION: The most common genetic mutations are --SEA for α-thalassemia and CD41-42 for ß-thalassemia in Liuzhou, Guangxi Zhuang Autonomous Region. A and ß-thalassemia can be detected at the same time by microarray chip technology in a high throughput manner.

Self-limiting electrospray deposition on polymer templates.

Electrospray deposition (ESD) applies a high voltage to liquids flowing through narrow capillaries to produce monodisperse generations of droplets down to hundreds of nanometers in diameter, each carrying a small amount of the delivered solute. This deposition method has been combined with insulated stencil masks for fabricating micropatterns by spraying solutions containing nanoparticles, polymers, or biomaterials. To optimize the fabrication process for micro-coatings, a self-limiting electrospray deposition (SLED) method has recently been developed. Here, we combine SLED with a pre-existing patterned polymer film to study SLED's fundamental behavior in a bilayer geometry. SLED has been observed when glassy insulating materials are sprayed onto conductive substrates, where a thickness-limited film forms as charge accumulates and repels the arrival of additional charged droplets. In this study, polystyrene (PS), Parylene C, and SU-8 thin films of varying thickness on silicon are utilized as insulated spraying substrates. Polyvinylpyrrolidone (PVP), a thermoplastic polymer is sprayed below its glass transition temperature (Tg) to investigate the SLED behavior on the pre-deposited insulating films. Furthermore, to examine the effects of in-plane confinement on the spray, a microhole array patterned onto the PS thin film by laser dewetting was sprayed with dyed PVP in the SLED mode. This was then extended to an unmasked electrode array showing that masked SLED and laser dewetting could be used to target microscale regions of conventionally-patterned electronics.

Analysis of rare thalassemia caused by HS-40 regulatory site deletion.

ABSTRACT Objectives: To investigate the effect of HS-40 regulatory site deletion on α-globin gene expression and its clinical significance. Methods: Venous blood samples of subjects were analyzed using a hematology analyzer and high- performance liquid chromatography; fetal cord blood was analyzed by a capillary electrophoresis analyzer. Gap-polymerase chain reaction (PCR), reverse dot blot (RDB), and multiple-link-dependent probe amplification (MLPA) were used for genotyping of thalassemia. Results: The proband was POLR3 K, HS-40 heterozygous deletion; the proband's wife was -SEA/αα; the fetus was POLR3 K, HS-40 heterozygous deletion combined with -SEA deletion; all of them had microcytic hypochromic anemia. Fetal umbilical cord blood electrophoresis revealed a suspected Hb Bart's band to be 88.4%, and the fetus was, thus, diagnosed as Hb Bart's fetus. The red blood cell parameters of the sporadic case showed that he had microcytic hypochromic anemia. Hemoglobin (Hb) electrophoresis analysis showed Hb H to be 5.3%, leading to a diagnosis of Hb H disease. Gap-PCR and RDB identified the genotype to be -α3.7/αα, ßA/ßA. MLPA detected heterozygous deletion or -α3.7 deletion on one allele and deletion of the HS-40 regulatory site on the other allele. Conclusion: The deletion of HS-40 regulatory site reduced expression of α-globin. HS-40 heterozygous deletion manifested as mild anemia, which was of microcytic hypochromic type. When compounded with -α3.7/αα, it manifested as Hb H disease; and when compounded with -SEA/αɑ, it manifested as Hb Bart's fetus.

Elevating EGFR-MAPK program by a nonconventional Cdc42 enhances intestinal epithelial survival and regeneration.

The regulatory mechanisms enabling the intestinal epithelium to maintain a high degree of regenerative capacity during mucosal injury remain unclear. Ex vivo survival and clonogenicity of intestinal stem cells (ISCs) strictly required growth response mediated by cell division control 42 (Cdc42) and Cdc42-deficient enteroids to undergo rapid apoptosis. Mechanistically, Cdc42 engaging with EGFR was required for EGF-stimulated, receptor-mediated endocytosis and sufficient to promote MAPK signaling. Proteomics and kinase analysis revealed that a physiologically, but nonconventionally, spliced Cdc42 variant 2 (V2) exhibited stronger MAPK-activating capability. Human CDC42-V2 is transcriptionally elevated in some colon tumor tissues. Accordingly, mice engineered to overexpress Cdc42-V2 in intestinal epithelium showed elevated MAPK signaling, enhanced regeneration, and reduced mucosal damage in response to irradiation. Overproducing Cdc42-V2 specifically in mouse ISCs enhanced intestinal regeneration following injury. Thus, the intrinsic Cdc42-MAPK program is required for intestinal epithelial regeneration, and elevating this signaling cascade is capable of initiating protection from genotoxic injury.

Rapid broadband discrete nanomechanical mapping of soft samples on atomic force microscope.

In this paper, an approach to achieve rapid broadband discrete nanomechanical mapping of soft samples using an atomic force microscope is developed. Nanomechanical mapping (NM) is needed to investigate, for example, dynamic evolution of the nanomechanical distribution of the sample-provided that the mapping is fast enough. The throughput of conventional NM methods, however, is inherently limited by the continuous scanning involved where the probe visits each sampling location continuously. Thus, we propose to significantly reduce the number of measurements through discrete mapping where only discrete sampling locations of interests are visited and measured. An online-searching learning-based technique is utilized to achieve rapid probe engagement and withdrawal with the interaction force minimized at each sampling location. Then, a control-based nanoindentation measurement technique is used to quickly acquire the nanomechanical property at each location, over frequencies that can be chosen arbitrarily in a broad range. Finally, a decomposition-based learning approach is explored to achieve rapid probe transitions between the sampling locations. The proposed technique is demonstrated through experiments using a Polydimethylsiloxane (PDMS) sample and a PDMS-epoxy sample as examples.

[Analysis of beta-globin gene variants in Liuzhou area of Guangxi].

OBJECTIVE: To determine the composition and distribution of beta-thalassemia-associated genotypes in Liuzhou area of Guangxi, China. METHODS: From January to December 2017, 13 847 individuals who came for premarital examination, maternity examination or health check were recruited with informed consent. The subjects were analyzed by reverse dot blotting (RDB) for 17 common beta-thalassemia-associated variants among the Chinese population. Individuals with inconsistent results by blood test, electrophoresis, and RDB were subjected to Sanger sequencing to detect rare variants of the beta globin gene. RESULTS: In total 2098 individuals were found to harbor beta-thalassemia-associated variants, which included 2075 heterozygotes (98.90%), 12 compound heterozygotes (0.57%) and 11 homozygotes (0.52%). CD41-42 (48.43%) and CD17 (31.45%) were the most common variants. Three hundred and thirty eight-individuals were found to also carry heterozygous variants of the alpha globin gene, with the most common types being --SEA/aa, -a3.7/aa, aCSa/aa, -a4.2/aa. Through Sanger sequencing, rare genotypes such as beta-32/betaN, betaCD41-42/betaIVS-II-5 and betaCD30/betaN were detected. CONCLUSION: Liuzhou area has a high incidence of beta-thalassemia, but with a complex variant spectrum and clinical phenotypes different from other regions. Genetic counseling and prenatal diagnosis for the carrier population is crucial for the reduction of the related birth defects. Our result may provide valuable information for the prevention and control of beta-thalassemia in this area.

Antibacterial TiCu/TiCuN Multilayer Films with Good Corrosion Resistance Deposited by Axial Magnetic Field-Enhanced Arc Ion Plating.

In order to develop a novel kind of antibacterial Cu-containing TiN film with good corrosion resistance, impressive mechanical properties, and low cytotoxicity, three differently designed multilayer films of TiCu/TiCuN multilayer (M1, M2, M3) were deposited on the surface of 316L stainless steel surface using the axial magnetic field-enhanced arc ion plating (AMFE-ARP) method, in which the interlayer of TiCu was first introduced for Cu-containing TiN film in order to improve comprehensive properties, especially the corrosion resistance of the film. The performance of the TiCu/TiCuN multilayer films was compared with that of the two single layers, TiN and TiCuN, which were deposited by the same method and the same total deposition time. The results indicated that the TiCu/TiCuN multilayer film of M2 revealed the best comprehensive corrosion resistance with low electric current values, high pitting potential, and high polarization resistance due to the proper thickness of TiCu interlayers and larger number of TiCu/TiCuN bilayers. In addition, the TiCu/TiCuN multilayer film of M2 also possesses comparable mechanical properties, excellent antibacterial and antibiofilm abilities, as well as good biocompatibility. Consequently, the antibacterial TiCu/TiCuN multilayer films with good corrosion resistance deposited by using the axial magnetic field-enhanced arc ion plating (AMFE-ARP) method are promising for application in biomedical antibacterial film for implants.

Receptor-mediated endocytosis generates nanomechanical force reflective of ligand identity and cellular property.

Whether environmental (thermal, chemical, and nutrient) signals generate quantifiable, nanoscale, mechanophysical changes in the cellular plasma membrane has not been well elucidated. Assessment of such mechanophysical properties of plasma membrane may shed lights on fundamental cellular process. Atomic force microscopic (AFM) measurement of the mechanical properties of live cells was hampered by the difficulty in accounting for the effects of the cantilever motion and the associated hydrodynamic force on the mechanical measurement. These challenges have been addressed in our recently developed control-based AFM nanomechanical measurement protocol, which enables a fast, noninvasive, broadband measurement of the real-time changes in plasma membrane elasticity in live cells. Here we show using this newly developed AFM platform that the plasma membrane of live mammalian cells exhibits a constant and quantifiable nanomechanical property, the membrane elasticity. This mechanical property sensitively changes in response to environmental factors, such as the thermal, chemical, and growth factor stimuli. We demonstrate that different chemical inhibitors of endocytosis elicit distinct changes in plasma membrane elastic modulus reflecting their specific molecular actions on the lipid configuration or the endocytic machinery. Interestingly, two different growth factors, EGF and Wnt3a, elicited distinct elastic force profiles revealed by AFM at the plasma membrane during receptor-mediated endocytosis. By applying this platform to genetically modified cells, we uncovered a previously unknown contribution of Cdc42, a key component of the cellular trafficking network, to EGF-stimulated endocytosis at plasma membrane. Together, this nanomechanical AFM study establishes an important foundation that is expandable and adaptable for investigation of cellular membrane evolution in response to various key extracellular signals.

[Study of closed colony of Microtus fortis infected with Schistosoma japonicum].

OBJECTIVE: To understand the pathological changes of tissues and organs in Microtus fortis infected with Schistosoma japonicum for further insight into the biological dynamic variation and explore immune mechanisms of M. fortis against S. japonicum infection. METHODS: The healthy M. fortis and mouse were infected with cercariae of S. japonicum through abdominal skin. On the day 12, 20 and 40, the M. fortis and mouse were sacrificed by cervical dislocation and dissected. The liver and kidney slices of M. fortis and mouse of the different groups were prepared and stained with hematoxylin eosin stain (HE) by conventional techniques, respectively. RESULTS: We did not find obvious lesions on the internal organs of mouse 12 d and 20 d after the infection and the lesions of M. fortis 40 d after the infection. However, white nodules appeared on the liver, kidney and spleen of the M. fortis 12 d and 20 d after the infection, and there were a lot of S. japonicum worms on pathological sections of inflamed liver and kidney tissues, and the boundary between worms and normal organization was clear. There were no parasites on pathological section of liver and kidney of mouse 12 d and 20 d after the infection. CONCLUSION: Among M. fortis, there are severe immune responses and the immune responses have an individual difference 12 days after S. japonicum infection.

Increasing manganese peroxidase production and biodecolorization of triphenylmethane dyes by novel fungal consortium.

A fungal consortium-SR consisting of Trametes sp. SQ01 and Chaetomium sp. R01 was developed for decolorizing three kinds of triphenylmethane dyes, which were decolorized by individual fungi with low efficiencies. The fungal consortium-SR produced 1.3 U ml(-1) of manganese peroxidase, 5.5 times higher than that produced by the monoculture of Trametes sp. SQ01, and decolorized Crystal Violet, Coomassie Brilliant Blue G250 (CBB G250) and Cresol Red. The fungal consortium-SR had a decolorization rate of 63-96%, much higher than that of the monoculture of strain SQ01 (38-72%). In consortium-SR, the higher efficiencies of decolorization of Crystal Violet and CBB G250 were obtained when they added to the culture after 4d of mixed cultivation rather than at the beginning of cultivation. Cresol Red was the exception. It is suggested that the consortium-SR has great potential for decolorizing triphenylmethane dyes.