Functional Polymorphisms of CTLA4 Associated with Aggressive Periodontitis in the Chinese Han Population.

BACKGROUND/AIMS: CTLA4 has been identified functioning as a protein receptor which functions as an immune checkpoint, downregulating the immune system. Susceptibility to aggressive periodontitis (AgP) is influenced by gene polymorphisms related to the immune response. In this study, we focused on SNPs in the 3'-UTR of CTLA4 among Chinese AgP patients, and investigated any further relationships between the SNPs and miRNAs. METHODS: This case-control study included 120 AgP patients and 150 healthy controls. Genotyping was used to detect allele distribution. Cell transfection and the dual luciferase reporter assay were performed to investigate the potential functions of SNPs located in the 3'UTR of CTLA4. RESULTS: The data show that patients with a history of smoking were more susceptible compared to controls, exhibiting deeper probing depth, greater attachment loss and more sites of bleeding on probing. The results of genotyping analysis revealed that individuals with the GA and AA genotypes, and with the A carrier had a decreased risk (P = 0.015, P = 0.03). Furthermore, patients with the G allele might be regulated by miR-105, which caused a down-regulation of CTLA4. The carriers of the GG genotype exhibited the worst results of attachment loss and bleeding on probing. CONCLUSION: These findings show that rs56102377 in the 3'-UTR of CTLA4 may act as a protective factor by disrupting the regulatory role of miR-105 in CTLA4 expression. Thus, our study highlighted a potential role of these polymorphisms as genetic susceptibility biomarkers of periodontitis in Chinese Han populations.

Synthesis and application of a targeting diagnosis system via quantum dots coated by amphiphilic polymer for the detection of liver cancer cells.

Water-soluble quantum dots (QDs) for liver cancer diagnosis were prepared using QDs with oleylamine ligand coated with poly(aspartate)-graft-poly(ethylene glycol)-dodecylamine (PASP-Na-g-PEG-DDA). Dynamic light scattering and transmission electron microscopy imaging showed that the novel QDs have an ellipsoidal morphology with a size of ~ 45 nm which could be used for biomedical application. Furthermore, the PASP-Na-g-PEG-DDA was then modified with anti-(vascular endothelial growth factor) (VEGF antibody), and a 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT) assay showed that the novel anti-VEGF-targeting QDs in vitro had low toxicity. Confocal laser scanning microscopy observations revealed an intracellular (HepG2) distribution of the novel anti-VEGF-targeting QDs and the targeting efficiency of anti-VEGF. These novel QDs could be used as a probe for liver cancer cell imaging because of anti-VEGF targeting.

Chitosan-coated nanoliposomes for the enhanced stability of walnut angiotensin-converting enzyme (ACE) inhibitory peptide.

This study encapsulated walnut angiotensin-converting enzyme (ACE) inhibitory peptides within nanoliposomes and then modified them with chitosan. The resulting effect of the nanoliposome loading and chitosan coating on physicochemical characteristics, stability, bioactivity, chemical structure, and morphology of the encapsulated peptides was assessed. The resulting particle size and polymer dispersity index revealed that the chitosan-coated nanoliposomes loaded with walnut ACE inhibitory peptides (WAIP) (CL-P) exhibited higher physical stability compared with the nanoliposomes loaded with WAIP (L-P). The encapsulation efficiency (EE) of CL-P increased from 73.32% to 76.13% after chitosan modification, and the EE of L-P and CL-P could be maintained by storage at 4°C. In addition, the antioxidant activity and ACE inhibitory activity of the peptides were effectively protected by L-P and CL-P during storage. Fourier transform infrared spectroscopy showed that the nanoliposomes were bound in ionic form with both the peptides and chitosan. Transmission electron micrographs indicated the presence of vesicle-like carriers with a reservoir-type structure. This study highlights the potential of nanoliposomes and their modification with chitosan to increase the stability and bioactivity retention of ACE inhibitory peptides. PRACTICAL APPLICATION: Chitosan-coated nanoliposomes loaded with walnut ACE inhibitory peptides were prepared in this study. Chitosan coating increased nanoliposomes' encapsulation efficiency and provided higher physical stability. In addition, the bioactivity of the walnut ACE peptides was effectively protected during storage. This study was relevant for improving the storage and transportation used for nanoliposome systems applied in the food and health product industry.

Acute myocardial infarction therapy using calycosin and tanshinone co-loaded; mitochondrion-targeted tetrapeptide and cyclic arginyl-glycyl-aspartic acid peptide co-modified lipid-polymer hybrid nano-system: preparation, characterization, and anti myocardial infarction activity assessment.

Acute myocardial infarction (AMI) is one of the most common ischemic heart diseases. However, lack of sufficient drug concentration (in the ischemic heart) is the major factor of treatment failure. It is urgent for researchers to engineer novel drug delivery systems to enhance the targeted delivery of cardioprotective agents. The aim of the present study was to investigate the anti-AMI ability of calycosin (CAL) and tanshinone (TAN) co-loaded; mitochondrion-targeted tetrapeptide (MTP) and cyclic arginyl-glycyl-aspartic acid (RGD) peptide co-modified nano-system.: We prepared CAL and TAN combined lipid-polymer hybrid nano-system, and RGD was modified to the system to achieve RGD-CAL/TAN NS. MTP-131 was conjugated with PEG and modified onto the nanoparticles to achieve dual ligands co-modified MTP/RGD-CAL/TAN NS. The physicochemical properties of nano-systems were characterized. The AMI therapy ability of the systems was investigated in AMI rats' model. The size of MTP/RGD-CAL/TAN NS was 170.2 ± 5.6 nm, with a surface charge of -18.9 ± 1.9 mV. The area under the curve (AUC) and blood circulation half-life (T1/2) of MTP/RGD-CAL/TAN NS was 178.86 ± 6.62 µg·min/mL and 0.47 h, respectively. MTP/RGD-CAL/TAN NS exhibited the most significant infarct size reduction effect of 22.9%. MTP/RGD-CAL/TAN NS exhibited the highest heart accumulation and best infarct size reduction effect, which could be used as a promising system for efficient treatment of cardiovascular diseases.

Acute myocardial infarction therapy using calycosin and tanshinone co-loaded mitochondria targeted lipid-polymer hybrid nano-system: Preparation, characterization, and anti myocardial infarction activity assessment.

BACKGROUND: Acute myocardial infarction (AMI) is one of the most common ischemic heart diseases. However, lack of sufficient drug concentrations in the ischemic heart may led to treatment failure. It is urgent for researchers to engineer novel drug delivery systems to enhance the targeted delivery of cardioprotective agents. OBJECTIVE: The aim of the present study was to investigate the anti-AMI ability of calycosin (CAL) and tanshinone (TAN) co-loaded mitochondria targeted lipid-polymer hybrid nano-system. METHODS: CAL and TAN combined lipid-polymer hybrid nano-systems were prepared and MTP-131 was conjugated with PEG and modified onto the nanoparticles to achieve MTP-CAL/TAN NS. The physicochemical properties of nano-systems were characterized, the AMI therapy ability of the systems was investigated in AMI rats' model. RESULTS: The size of MTP-CAL/TAN NS was 168.7 ±â€¯5.1 nm, with a surface charge of - 21.3 ±â€¯2.3 mV. The area under the curve (AUC) and blood circulation half-life (T1/2) of MTP-CAL/TAN NS was 178.86 ±â€¯6.62 µg·min/mL and 0.47 h, respectively. MTP-CAL/TAN NS exhibited the most significant infarct size reduction effect of 23.9 %. CONCLUSION: MTP-CAL/TAN NS exhibited the highest heart accumulation and best infarct size reduction effect, which could be used as a promising system for efficient treatment of cardiovascular diseases.

ROS-sensitive calcipotriol nano-micelles prepared by methoxypolyethylene glycol (mPEG) - modified polymer for the treatment of psoriasis.

Oxidative stress due to excessive reactive oxygen species (ROS) production in the skin microenvironment is one of the main mechanisms in psoriasis pathogenesis. A nano drug delivery system based on ROS-responsive release can enhance drug release at the target site. In this study, a ROS-sensitive material methoxypolyethylene glycol-thioether-thiol (mPEG-SS) was synthesized using mPEG as the parent structure with sulfide structural modification. An mPEG-SS-calcipotriol (mPEG-SS-CPT, PSC) nano-micelle percutaneous delivery system was prepared by encapsulating CPT. A small animal imaging system was used to study PSC's the ROS-sensitive drug release process. It is shown that endogenous ROS mainly affects PSC and releases drugs. Finally, the therapeutic effect of PSC on psoriasis was explored by animal experiments. Ultimately, it ameliorates imiquimod-induced psoriasis-like inflammation. Overall, PSC is an effective ROS-sensitive transdermal drug delivery system that is expected to provide a new strategy for treating psoriasis.

Vanadium recovery by electrodialysis using polymer inclusion membranes.

Industrial applications and environmental awareness recently prompted vanadium recovery spell from secondary resources. In this work, a polymer inclusion membrane containing trioctylmethylammonium chloride as carrier was successfully employed in electrodialysis for vanadium recovery from acidic sulfate solutions. The permeability coefficient of V(V) increased from 0.29 µm·s-1 (without electric field) to 4.10 µm·s-1 (with the 20 mA·cm-2 current density). The transport performance of VO2SO4-, which was the predominant species containing V(V) in the acidic region (pH <3), was influenced by the aqueous pH value and sulfate concentration. Under an electric field, a low concentrated H2SO4 solution (0.2 M) effectively stripped V(V) from the membranes, avoiding the requirement of a highly concentrated H2SO4 without electric field. Under the optimum conditions, the permeability coefficient and flux reached 6.80 µm·s-1 and 13.34 µmol·m-2·s-1, respectively. High selectivity was observed for the separation of V(V) and Mo(VI) from mixed solutions of Co (II), Ni (II), Mn (II), and Al (III). Additionally, the separation between Mo(VI) and V(V) was further improved by adjusting the acidity of the stripping solution. The V(V) selectivity for the resulting membrane was higher than that of commercial anion exchange membranes.

Ultrasound-Assisted Preparation of Exopolysaccharide/Nystatin Nanoemulsion for Treatment of Vulvovaginal Candidiasis.

PURPOSE: As one of the classic anti-Canidia albicans (CA) and vulvovaginal candidiasis (VVC) drugs, nystatin (NYS) is limited by poor water solubility and easy aggregation. Traditional NYS vaginal delivery formulations do not fully adapt to the specific environment of the vaginal cavity. The use of exopolysaccharides (EPS) has great application potential in emulsifiers, but its use has not been reported in nanoemulsions. In this work, an EPS/NYS nanoemulsion (ENNE) was developed to improve the activities of NYS against CA and VVC. METHODS: The ENNE was prepared by ultrasonic method using EPS as an emulsifier, liquid paraffin oil as an oil phase, PEG400 as a co-emulsifier, and NYS as the loaded drug. ENNE preparation was optimized by response surface method. After optimization, in vitro and in vivo analysis of the anti-CA activity; animal experiments; staining with propidium iodide (PI), periodic acid-schiff (PAS), and hematoxylin-eosin (H&E); and cytokine experiments were performed to investigate the therapeutic ability against VVC. RESULTS: The optimal formulation and preparation parameters of ENNE were determined as follows: EPS content of 1.5%, PEG400 content of 3.2%, NYS content of 700 µg/mL, paraffin oil content of 5.0%, ultrasonic time of 15 min, and ultrasonic amplitude of 35%. The ENNE showed an encapsulated structure with an average particle size of 131.1 ± 4.32 nm. ENNE exhibited high storage and pH stability, as well as slow release. The minimum inhibitory concentration (MIC) of ENNE against CA was only 0.125 µg/mL and the inhibition zone was 19.0 ± 0.5 mm, for greatly improved anti-CA effect. The prepared ENNE destroyed the membrane of CA cells, and exhibited good anti-CA effect in vivo and therapeutic ability against VVC. CONCLUSION: The results of this study will promote the application of EPS in nanotechnology, which should lead to new and effective local drug formulations for treating VVC.

Co-delivery of docetaxel and curcumin prodrug via dual-targeted nanoparticles with synergistic antitumor activity against prostate cancer.

PURPOSE: Combination therapy is increasingly used as a primary cancer treatment regimen. In this report, we designed EGFR peptide decorated nanoparticles (NPs) to co-deliver docetaxel (DTX) and pH sensitive curcumin (CUR) prodrug for the treatment of prostate cancer. RESULTS: EGFR peptide (GE11) targeted, pH sensitive, DTX and CUR prodrug NPs (GE11-DTX-CUR NPs) had an average diameter of 167nm and a zeta potential of -37.5mV. The particle size of the NPs was adequately maintained in serum and a sustained drug release pattern was observed. Improved inhibition of cancer cell and tumor tissue growth was shown in the GE11-DTX-CUR NPs group compared to the other groups. CONCLUSION: It can be summarized that DTX and CUR prodrug could be delivered into tumor cells simultaneously by the GE 11 targeting and the EPR effect of NPs. The resulting GE11-DTX-CUR NPs is a promising system for the synergistic antitumor treatment of prostate cancer.

Targeted nanomedicine for prostate cancer therapy: docetaxel and curcumin co-encapsulated lipid-polymer hybrid nanoparticles for the enhanced anti-tumor activity in vitro and in vivo.

OBJECTIVE: Docetaxel (DTX) remains the only effective drug for prolonging survival and improving quality of life of metastatic castration-resistant prostate cancer (mCRPC) patients. Combination anticancer therapy encapsulating DTX and another extract of traditional Chinese medicine is one nano-sized drug delivery system promising to generate synergistic anticancer effects, to maximize the treatment effect, and to overcome multi-drug resistance. The purpose of this study is to construct lipid-polymer hybrid nanoparticles (LPNs) as nanomedicine for co-encapsulation of DTX and curcumin (CUR). METHODS: DTX and CUR co-encapsulated LPNs (DTX-CUR-LPNs) were constructed. DTX-CUR-LPNs were evaluated in terms of particles size, zeta potential, drug encapsulation, and drug delivery. The cytotoxicity of the LPNs was evaluated on PC-3 human prostate carcinoma cells (PC3 cells) by MTT assays. In vivo anti-tumor effects were observed on the PC3 tumor xenografts in mice. RESULTS: The particle size of DTX-CUR-LPNs was 169.6 nm with a positive zeta potential of 35.7 mV. DTX-CUR-LPNs showed highest cytotoxicity and synergistic effect of two drugs in tumor cells in vitro. In mice-bearing PC-3 tumor xenografts, the DTX-CUR-LPNs inhibited tumor growth to a greater extent than other contrast groups, without inducing any obvious side effects. CONCLUSION: According to these results, the novel nanomedicine offers great promise for the dual drugs delivery to the prostate cancer cells, showing the potential of synergistic combination therapy for prostate cancer.

In-Situ-Generated Vasoactive Intestinal Peptide Loaded Microspheres in Mussel-Inspired Polycaprolactone Nanosheets Creating Spatiotemporal Releasing Microenvironment to Promote Wound Healing and Angiogenesis.

Vasoactive intestinal peptide (VIP) was reported to promote angiogenesis. Electrospun nanofibers lead to idea wound dressing substrates. Here we report a convenient and novel method to produce VIP loaded microspheres in polycaprolactone (PCL) nanofibrous membrane without complicated processes. We first coated mussel-inspired dopamine (DA) to nanofibers, then used strong adhesive DA to absorb the functional peptide. PCL membrane was then immersed into acetone to generate microspheres with VIP loading. We employed high pressure liquid chromatography to record encapsulation efficiency of (31.8 ± 2.2)% and loading capacity of (1.71 ± 0.16)%. The release profile of VIP from nanosheets showed a prolonged release. The results of laser scanning confocal microscope, scanning electron microscope and cell counting kit-8 proliferation assays showed that cell adhesion and proliferation were promoted. In order to verify the efficacy on wound healing, in vivo implantation was applied in the full-thickness defect wounds of BALB/c mice. Results showed that the wound healing was significantly promoted via favoring the growth of granulation tissue and angiogenesis. However, we found wound re-epithelialization was not significantly improved. The resulting VIP-DA-coated PCL (PCL-DA-VIP) nanosheets with spatiotemporal delivery of VIP could be a potential application in wound treatment and vascular tissue engineering.

Controlled water vapor transmission rate promotes wound-healing via wound re-epithelialization and contraction enhancement.

A desirable microenvironment is essential for wound healing, in which an ideal moisture content is one of the most important factors. The fundamental function and requirement for wound dressings is to keep the wound at an optimal moisture. Here, we prepared serial polyurethane (PU) membrane dressings with graded water vapor transmission rates (WVTRs), and the optimal WVTR of the dressing for wound healing was identified by both in vitro and in vivo studies. It was found that the dressing with a WVTR of 2028.3 ± 237.8 g/m(2)·24 h was able to maintain an optimal moisture content for the proliferation and regular function of epidermal cells and fibroblasts in a three-dimensional culture model. Moreover, the dressing with this optimal WTVR was found to be able to promote wound healing in a mouse skin wound model. Our finds may be helpful in the design of wound dressing for wound regeneration in the future.

Novel bilayer wound dressing composed of silicone rubber with particular micropores enhanced wound re-epithelialization and contraction.

Wound dressing is critical important for cutaneous wound healing. However, the application of current products is limited due to poor mechanical property, unsuitable water vapor transmission rate (WVTR), poor anti-infective property or poor biocompatibility, etc. In the present study, a microporous silicone rubber membrane bilayer (SRM-B) composed of two layers with different pore sizes was prepared. The physical properties, the influences of pore structure on the bacterial penetration, the cell adhesion and proliferation were studied. Lastly, the effects of the SRM-B on the healing of a mouse full-thickness wound were examined. The data showed that the small pore upper layer of SRM-B could effectively prevent the bacterial invasion, as well as properly keep the water vapor transmission rate; the large pore lower layer of SRM-B could promote the cell adhesion and proliferation. The in vivo results showed that SRM-B could significantly enhance wound re-epithelialization and contraction, which accelerated the wound healing. Our data suggested that the SRM-B, with different particular pore sizes, could serve as a kind of promising wound dressing.

Cytotoxicity of PEGylated graphene oxide on lymphoma cells.

Graphene oxide (GO) is a hotspot, especially in the field of biomedical. However, the clinical application of GO is still faces a lot of challenges. In order to improve the solubility and biocompatibility of GO, polyethylene glycol (PEG) was grafted on the surface of graphene oxide by amide reaction. PEGylated graphene oxide (PEG-GO) was characterized using Fourier transform infrared spectroscopy (FTIR). The stability of PEG-GO detected in different solutions. Raji cell was selected as a lymphoma cell model to study the cytotoxicity of PEG-GO. Cell viability was detected using the Cell Counting Kit-8 assay. Cells were treated with different concentrations (10-100 µg/mL) of PEG-GO at different time points (6, 12, and 24 h). The FTIR spectrum of PEG-GO indicated that polyethylene glycol was successfully grafted onto GO. PEG-GO had excellent stability in all solutions. Cells treated with PEG-GO (10-100 µg/mL) for 24 hours had survival rates were over 80%. These results demonstrate that PEG-GO had an excellent dispersion in biological solutions and the toxicity of PEG-GO to lymphoma cells was low. The paper may provide cytological evidence for the application of PEG-GO in medicine.

A smart drug delivery system from charge-conversion polymer-drug conjugate for enhancing tumor therapy and tunable drug release.

A smart drug delivery system is prepared by citraconylated polyaspartic acid (PASP) derivate-drug conjugate. The conjugate contains two pH-sensitive groups: citraconic amide and hydrazone linker. Citraconic amide group can enhance tumor therapy efficiency by the extracellular pH-sensitive charge-conversion property. Hydrazone linker between polymer and drug can cleave efficiently in the intracellular pH environment. The resulting conjugate shows dual-pH sensitive properties: extracellular pH-triggered enhanced tumor targeting and intracellular pH-triggered drug release. The results of physicochemical properties, intracellular location, and cytotoxicity of conjugate micelles demonstrate that this novel smart drug delivery system can enhance intracellular delivery of drug at a low pH and then release drug rapidly.

[Clinical study on topical bismuth subgallate/borneol (Suile) dressing for treatment of diabetic foot ulcers].

OBJECTIVE: To evaluate the efficacy and safety of Bismuth Subgallate/Borneol (Suile) (BSB) dressing in the treatment of diabetic foot ulcers. METHODS: A two-center, randomized controlled parallel-group comparison study was conducted. Between September 2005 and November 2006, 35 patients with nonhealing diabetic foot ulcer (Wagner 2-3 grade) were recruited and divided randomly into the test group (BSB group, n = 25) and control group (IG group, n = 10). There was no significant difference in general data between 2 groups (P > 0.05). Based on the comprehensive therapy of diabetic foot, ulcers were topically treated by Suile dressing and Intrasite gel in the BSB group and IG group, respectively. The ulcer area was measured once a week. Ulcer bleeding was observed and acceptance of the dressing was inquired each week. RESULTS: Of the patients, 22 patients in the BSB group and 8 in the IG group completed the clinical trial. Intention to treat (ITT) analysis indicated that the results were excellent in 19 (76%) cases, good in 3 cases (12%), fair in 1 case (4%), and poor in 2 cases (8%) in the BSB group. In the IG group, the results were excellent in 8 cases (80%) cases, fair in 1 case (10%), and poor in 1 case (10%). Per-protocol (PP) analysis showed that the results were excellent in 19 cases (86%) and good in 3 cases (14%) in the BSB group, and were excellent in 8 cases (100%) in the IG group. The results of ITT and PP analysis all showed no significant difference between 2 groups (P > 0.05). The change trend of ulcer areas in the BSB group was similar to that in the IG group. There was no significant difference in the hemostatic effect between 2 groups (P > 0.05) and the patients were more likely to accept BSB dressings. CONCLUSION: BSB dressings is an effective, safe, and generally well-tolerated therapy dressing in the treatment of diabetic foot ulcers.

Induction of apoptosis and cell cycle arrest by polyvinylpyrrolidone K-30 and protective effect of alpha-tocopherol.

Polyvinylpyrrolidone is a macromolecular polymer with widespread use in industry as well as in medicine for various purposes. Its effect on cells cultured in vitro, however, has not been fully investigated. To elucidate this issue, we studied the influence of PVP K-30 on cultured HeLa cells. PVP K-30 treatment produced a dose- and time-dependent toxicity to HeLa cells. Cells exposed to PVP K-30 exhibited several morphological features of apoptosis. Gel electrophoresis of DNA from PVP K-30-treated cells showed typical apoptotic ladder. And flow cytometric analysis demonstrated that PVP K-30 induced cell cycle arrest at G2/M phase and the subsequent appearance of sub-G1 population. In addition, it was shown that procaspase-3 was activated in response to PVP K-30 treatment. We also found that alpha-tocopherol efficiently protected HeLa cells from PVP K-30 cytotoxicity. This is the first demonstration that PVP K-30 could induce apoptosis in HeLa cells and cell cycle arrest at G2/M phase, and that PVP K-30 toxicity could be attenuated by alpha-tocopherol.