Multiple miRNA Detection through a Suspended Microbead Array Encoded by Triple-Color Upconversion Luminescent Nanotags via Bi-Beam Splitter Hybrid-Multitrap Optical Tweezers.

In recent years, optical tweezers have become a novel tool for biodetection, and to improve the inefficiency of a single trap, the development of multitraps is required. Herein, we constructed a set of hybrid multitrap optical tweezers with the balance of stability and flexibility by the combination of two different beam splitters, a diffraction optical element (DOE) and galvano mirrors (GMs), to capture polystyrene (PS) microbeads in aqueous solutions to create an 18-trap suspended array. A sandwich hybridization strategy of DNA-miRNA-DNA was adopted to detect three kinds of target miRNAs associated with triple negative breast cancer (TNBC), in which different upconversion nanoparticles (UCNPs) with red, green, and blue emissions were applied as luminescent tags to encode the carrier PS microbeads to further indicate the levels of the targets. With encoded luminescent microbeads imaged by a three-channel microscopic system, the biodetection displayed high sensitivity with low limits of detection (LODs) of 0.27, 0.32, and 0.33 fM and exceptional linear ranges of 0.5 fM to 1 nM, 0.7 fM to 1 nM, and 1 fM to 1 nM for miR-343-3p, miR-155, and miR-199a-5p, respectively. In addition, this bead-based assay method was demonstrated to have the potential for being applied in patients' serum by satisfactory standard addition recovery experiment results.

The impact of smoking on periodontitis patients' GCF/serum cytokine profile both before and after periodontal therapy: a meta-analysis.

BACKGROUND: Smoking is an established modifying factor for the host immune response of periodontitis patients. However, its exact influence remains unclear. We aimed to compare the cytokine profile of periodontitis patients with and without smoking habits both before and after periodontal therapy to preliminarily explore its influence on the host immune response to periodontitis. METHODS: The protocol of the present meta-analysis was registered in the International Prospective Register of Systematic Reviews (PROSPERO) under the code CRD42021255656. Meta-analysis was performed for each cytokine if at least three studies were included. We synthesized the evidence to compare the cytokine profile of periodontitis with and without smoking both in gingival cervical fluid (GCF) and serum to explore the impact of smoking on periodontitis both locally and systemically. Moreover, we also compared the cytokine profile of the two groups of patients after periodontal therapy to explore the effect of smoking on the outcome of periodontal therapy. RESULTS: Fifteen studies were included in this meta-analysis. We found that there was no significant difference between the two groups of patients in the baseline cytokine profile. However, after periodontal therapy, smoking periodontitis patients showed significantly higher IL-1ß levels in their GCF than nonsmoking patients. DISCUSSION: There was no significant difference between smoking and nonsmoking periodontitis patients in the baseline cytokine profile. However, after periodontal therapy, smoking periodontitis patients showed significantly higher IL-1ß levels in their GCF than nonsmoking patients, which indicates that smoking may impair the response of periodontitis to periodontal treatment.

A magnetic protein biocompass.

The notion that animals can detect the Earth's magnetic field was once ridiculed, but is now well established. Yet the biological nature of such magnetosensing phenomenon remains unknown. Here, we report a putative magnetic receptor (Drosophila CG8198, here named MagR) and a multimeric magnetosensing rod-like protein complex, identified by theoretical postulation and genome-wide screening, and validated with cellular, biochemical, structural and biophysical methods. The magnetosensing complex consists of the identified putative magnetoreceptor and known magnetoreception-related photoreceptor cryptochromes (Cry), has the attributes of both Cry- and iron-based systems, and exhibits spontaneous alignment in magnetic fields, including that of the Earth. Such a protein complex may form the basis of magnetoreception in animals, and may lead to applications across multiple fields.

The osseointegration and stability of dental implants with different surface treatments in animal models: a network meta-analysis.

Dental implants are commonly used to repair missing teeth. The implant surface plays a critical role in promoting osseointegration and implant success. However, little information is available about which implant surface treatment technology best promotes osseointegration and implant stability. The aim of this network meta-analysis was to evaluate the osseointegration and stability of four commonly used dental implants (SLA, SLActive, TiUnite, and Osseotite). The protocol of the current meta-analysis is registered in PROSPERO (International Prospective Register of Systematic Reviews) under the code CRD42020190907 ( https://www.crd.york.ac.uk ). We conducted a systematic review following PRISMA and Cochrane Recommendations. Medline (PubMed), Cochrane Library, Embase, and the Web of Science databases were searched. Only randomized controlled trials were considered. Twelve studies were included in the current network meta-analysis, eleven studies were included concerning the osseointegration effect and five studies were included for stability analysis (four studies were used to assess both stability and osseointegration). Rank possibility shows that the SLActive surface best promoted bone formation at an early healing stage and TiUnite seemed to be the best surface for overall osseointegration. For stability, TiUnite seemed to be the best surface. The present network meta-analysis showed that the SLActive surface has the potential to promote osseointegration at an early stage. The TiUnite surface had the best effect on osseointegration regarding the overall healing period. The TiUnite surface also had the best effect in stability.

Gradient nanostructured titanium stimulates cell responses in vitro and enhances osseointegration in vivo.

BACKGROUND: Though titanium (Ti) is widely used as dental materials in the clinic, effective methods to treat Ti for higher surface biological activity still lack. Through Surface mechanical attrition treatment (SMAT) technology we could endow Ti with gradient nanostructured surface (GNS Ti). To investigate the biocompatibility of GNS Ti for its further application in dental implant field, we study the effects of GNS Ti on cell responses in vitro and osseointegration of the implant with surrounding bone tissues in vivo. METHODS: In this study, GNS Ti was fabricated by SMAT. In vitro experiment, we co-cultured GNS Ti with bone mesenchymal stem cells (BMSCs), surface characterization was detected by transmission electron microscope (TEM). Adhesion, proliferation and differentiation of BMSCs were evaluated by scanning electron microscope (SEM), MTT, flow cytometry (FCM), alkaline phosphatase (ALP) and osteocalcin (OCN) tests. In vivo experiment, the GNS Ti was implanted into the rabbit mandible. Osteogenesis and osseointegration were evaluated by Micro CT, toluidine blue staining, and immunohistochemical staining at 4, 8, and 12 weeks postoperatively. RESULTS: Both results showed that compared with the coarse grained (CG) Ti, the GNS Ti stimulated the adhesion, proliferation, and differentiation of BMSCs and improved osteogenesis and osseointegration. CONCLUSIONS: This study indicates that gradient nanostructured Ti is a promising material for dental implant application.

Characterization of different biodegradable scaffolds in tissue engineering.

The aim of the present study was to compare the characteristics of acellular dermal matrix (ADM), small intestinal submucosa (SIS) and Bio­Gide scaffolds with acellular vascular matrix (ACVM)­0.25% human­like collagen I (HLC­I) scaffold in tissue engineering blood vessels. The ACVM­0.25% HLC­I scaffold was prepared and compared with ADM, SIS and Bio­Gide scaffolds via hematoxylin and eosin (H&E) staining, Masson staining and scanning electron microscope (SEM) observations. Primary human gingival fibroblasts (HGFs) were cultured and identified. Then, the experiment was established via the seeding of HGFs on different scaffolds for 1, 4 and 7 days. The compatibility of four different scaffolds with HGFs was evaluated by H&E staining, SEM observation and Cell Counting Kit­8 assay. Then, a series of experiments were conducted to evaluate water absorption capacities, mechanical abilities, the ultra­microstructure and the cytotoxicity of the four scaffolds. The ACVM­0.25% HLC­I scaffold was revealed to exhibit the best cell proliferation and good cell architecture. ADM and Bio­Gide scaffolds exhibited good mechanical stability but cell proliferation was reduced when compared with the ACVM­0.25% HLC­I scaffold. In addition, SIS scaffolds exhibited the worst cell proliferation. The ACVM­0.25% HLC­I scaffold exhibited the best water absorption, followed by the SIS and Bio­Gide scaffolds, and then the ADM scaffold. In conclusion, the ACVM­0.25% HLC­I scaffold has good mechanical properties as a tissue engineering scaffold and the present results suggest that it has better biological characterization when compared with other scaffold types.

Expanding the clinical and genetic spectrum of Heimler syndrome.

BACKGROUND: Heimler syndrome (HS) is a rare hereditary systemic disorder, partial clinically overlapping with Usher syndrome. So far, our knowledge of HS is very limited, many cases are misdiagnosed or may not even be diagnosed at all. This study aimed to analyze the clinical and genetic characteristics of HS, and to evaluate potential phenotype-genotype correlations. RESULTS: Two HS cases caused by PEX1 mutations were identified, and a novel likely pathogenic mutation, PEX1 c.895_896insTATA, was found. The main ophthalmic finding of the two patients was consistent with retinitis pigmentosa accompanied by cystoid macular edema, but short axial length and hyperopia were also observed as two previously unreported ocular phenotypes. Analysis of the literature showed that of the 29 HS patients previously reported, 12 had PEX6 mutations, 10 had PEX1 mutations, two had PEX26 mutations, and the remaining patients were not genetically tested. Three novel genotype-phenotype correlations were revealed from analysis of these patients. First, most genotypes of every HS patient include at least one missense variant; second, at least one mutation in PEX1 or PEX6 gene affects the AAA-ATPase region in every HS patient with retinal dystrophy, suggesting AAA-ATPase region is a hypermutable region in patients with a retinal dystrophy; third, there are no significant differences between PEX1-, PEX6-, and PEX26-associated phenotypes. CONCLUSION: Next-generation sequencing is important for the diagnosis of HS. This study expands the clinical and genetic spectrum of HS, and provides additional insights into genotype-phenotype correlations, which is vital for accurate clinical practice, genetic counseling, and pathogenesis studies.

Protein preparation, crystallization and preliminary X-ray crystallographic analysis of Smu.1475c from caries pathogen Streptococcus mutans.

The gene smu.1475c encodes a putative protein of 211 residues in Streptococcus mutans, a primary pathogen for human dental caries. In this work, smu.1475c was cloned into pET28a and expressed in good amount from the E. coli strain BL21 (DE3). Smu.1475c protein was purified to homogeneity in a two-step procedure of Ni2+ chelating and size exclusion chromatography. Crystals were obtained by hanging-drop vapor-diffusion method and diffracted to 2.7 angstroms resolution. The crystal belongs to orthorhombic space group P2(1)2(1)2(1) with cell dimension of a = 68.3 angstroms, b = 105.9 angstroms, c = 136.2 angstroms. The asymmetric unit is expected to contain four molecules with solvent content of 49.4%.

Protein preparation, crystallization and preliminary X-ray crystallographic analysis of SMU.961 protein from the caries pathogen Streptococcus mutans.

The smu.961 gene encodes a putative protein of 183 residues in Streptococcus mutans, a major pathogen in human dental caries. The gene was cloned into expression vector pET28a and expressed in a substantial quantity in Escherichia coli strain BL21 (DE3) with a His tag at its N-terminus. The recombinant protein SMU.961 was purified to homogeneity in a two-step procedure consisting of Ni2+-chelating and size-exclusion chromatography. Crystals suitable for X-ray diffraction were obtained by the hanging-drop vapour-diffusion method and diffracted to 2.9 A resolution at beamline I911-3, MAX-II-lab, Sweden. The crystal belonged to space group C2, with unit-cell parameters a = 98.62, b = 73.73, c = 184.73 A, beta = 98.82 degrees.

Crystallization and preliminary X-ray crystallographic studies of SMU.134 protein from caries pathogen Streptococcus mutans.

The smu.134 gene encodes a putative transcriptional regulator of 217 residues in Streptococcus mutans, a major pathogen for human dental caries. The gene was cloned into expression vector pET28alpha and expressed in soluble form in E. coli strain BL21 (DE3) with a His tag at its N-terminus. The recombinant protein SMU.134 was purified to homogeneity in a two step procedure of Ni(2+ ) chelating and size exclusion chromatography. Crystals suitable for X-ray diffraction were obtained by hanging-drop vapor diffusion method and diffracted to 2.6 A;. The crystal belonged to space group P2(1)2(1)2(1), with unit-cell parameters a=55.03 A, b=80.84 A, c=107.96 A.

Comprehensive analysis of interleukin-8 gene polymorphisms and periodontitis susceptibility.

BACKGROUND: Associations between interleukin-8 (IL-8) gene polymorphisms and periodontitis susceptibility have been investigated in many published studies, but the conclusions are still inconsistent. Therefore, we performed this systematic review and meta-analysis to review which polymorphisms have been researched and to obtain a precise result of the same polymorphism from different studies. RESULTS: Finally 10 publications involving 1938 patients and 1569 controls were yielded, including 12 polymorphisms. Six studies investigated rs4073 polymorphism; two focused on rs2227306 and rs2227307; two referred to rs2227532 and T-738A; one detected rs2230054, rs1126579 and rs1126580; one inspected A2767T, T11722T2 and C1633T, and one for rs2234671 polymorphism. Of them, IL-8 C1633T and rs1126580 polymorphisms showed positive association while the other ten polymorphisms revealed negative results. MATERIALS AND METHODS: A comprehensive literature search from PubMed, Web of Science, and Chinese National Knowledge Infrastructure was conducted for all potentially relevant studies published before January 2, 2017. Two authors selected the studies and extracted data. The pooled analysis was conducted using the RevMan 5.1 software if a polymorphism was reported by two or more studies. CONCLUSIONS: Based on current evidence, the IL-8 rs4073, A2767T, T11722T2, rs2234671, rs2230054, rs1126579, rs2227306, rs2227307, rs2227532, and T-738A polymorphisms were not associated with periodontitis susceptibility; the IL-8 C1633T and rs1126580 polymorphisms were associated with increased risk of periodontitis.

Effects of additives and processing parameters on the initial burst release of protein from poly(lactic-co-glycolic acid) microspheres.

The aim of this study is to investigate both the effects of hydrophilic additives and combined processing parameters on the in vitro release of a model protein, bovine serum albumin (BSA), from poly(lactic-co-glycolic acid) (PLGA) microspheres. Additives including beta-cyclodextrin, HP-beta-cyclodextrin, poly(ethylene glycol) (PEG) 6000, and sorbitol, and processing parameters such as the poly(vinyl alcohol) (PVA) concentration, emulsification temperature, aqueous/oil phase, evaporation method, and dehydration method were evaluated. PLGA microspheres were all prepared by the double-emulsion solvent extraction/evaporation method, and the results showed that no statistically significant differences of particle sizes and entrapment efficiencies appeared. Interestingly, the initial burst releases were markedly changed by both additives and processing parameters. Initial burst releases were accelerated by hydrophilic additives except for PEG 6000 and were retarded by the formulation composed of higher PVA concentration, tween-20 as an emulsifier in the internal aqueous phase, glycerol in the oil phase, and inorganic salt in the external aqueous phase, and operated at low temperature. Scanning electron microscopy showed that the more porous and dimpled the structure on the surface of the PLGA microspheres, the larger the initial burst release. The microspheres that displayed a relatively smooth and compact surface showed the least burst release.

[Preparation of alginate-chitosan-poly (lactic-co-glycolic acid) composite microsphere and its regulation of protein release].

AIM: To elevate the encapsulation efficiency, decrease the burst release and improve the release of protein entrapped in poly (lactic-co-glycolic acid) (PLGA) microspheres. The bovine serum albumin (BSA) composite microspheres of alginate-chitosan-PLGA were prepared and the release characteristics of BSA from this composite microspheres were studied. METHODS: The much smaller calcium alginate microcapsules were first prepared by a modified emulsification method in an isopropyl alcohol-washed way and coated with chitosan, then the alginate-chitosan microcapsules were further entrapped in PLGA to form the composite microspheres. The protein concentration was determined using a BCA protein assay kit. The release profiles were changed with various formulation factors. RESULTS: The average diameter of the composite microcapsules was about 30 microm. Comparing with 60% to 70% of the conventional PLGA microspheres, the average encapsulation efficiency was more than 80%, and the burst releases in phosphate buffer solution of the composite microspheres decreased from 40% and 50% to 25% and further to 5% in saline solution. CONCLUSION: The novel composite microspheres were prepared, the drug encapsulation efficiency increased and the burst release decreased. The desired release profiles could be obtained by regulating the ratios of PLG and PLA in the composite microspheres.

[The influence of Co-Cr-Mo alloys on biological behaviour of L929 mouse fibroblasts].

PURPOSE: To investigate the influence of dental Co-Cr-Mo alloys on biological behaviour of L929 mouse fibroblasts. METHODS: Leaching liquor of medical pure titanium, Co-Cr-Mo alloys, Co-Cr alloys and Ni-Cr alloys was prepared and used to culture the L929 cells for 24 h, 48 h and 72 h, respectively; then the growth of L929 cells was observed under inverted phase contrast microscope, and the cytotoxicity grades of 4 kinds of materials were evaluated by using CCK-8 test. The apoptosis of L929 cells was measured by flow cytometry(FCM) and acridine orange staining was used to observe the L929 cells adhered on the surface of samples under fluorescent microscope. The data was statistically analyzed with SPSS17.0 software package. RESULTS: Under inverted phase contrast microscope, at each time point, L929 cells grew in a good condition except in the Ni-Cr alloys group that minor karyopyknosis was indicating slight celluar cytotoxicity. Optical density (OD), apoptosis rate and cell adhesion number in Co-Cr-Mo alloys group at each time point were significantly less than in medical pure titanium group (P<0.05), and higher than in Co-Cr alloys group (P<0.05) and Ni-Cr alloys group (P<0.05). During observation period, the cytotoxicity grades of medical pure titanium group, Co-Cr-Mo alloys group and Co-Cr alloys group was grade 1, while Ni-Cr alloy group was grade 2, i.e. mild cytotoxicity. CONCLUSIONS: Co-Cr-Mo alloys have no adverse effect on the biological behavior of L929 cells, which may be suitable for dental clinical application with good biocompatibility.

Preparation and characteristics of lipid nanoemulsion formulations loaded with doxorubicin.

PURPOSE: Safe and effective lipid nanoemulsion (LNE) formulations for the antitumor delivery of doxorubicin is designed. METHODS: LNEs composed of medium-chain triglyceride, soybean oil, lecithin, and doxorubicin are prepared by a solvent-diffusion method in an aqueous system. The effects of lipid material composition and polyethylene glycol (PEG)ylation on the size, drug encapsulation efficiency, and stability of LNEs are investigated. Based on in-vitro cytotoxicity and cellular uptake tests of A549 (human lung carcinoma) cells, in-vivo biodistribution, antitumor activity, and cardiac toxicity are further examined using nude mouse bearing A549 tumor. RESULTS: The LNE size decreases from 126.4 ± 8.7 nm to 44.5 ± 9.3 nm with increased weight ratio of medium-chain triglyceride to soybean oil from 1:4 to 3:2, whereas the encapsulation efficiency of doxorubicin is slightly reduced from 79.2% ± 2.1% to 71.2% ± 2.9%. The PEGylation of LNE by 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[carboxy(PEG)2000] (DSPE-PEG 2000) does not significantly change the size and drug encapsulation efficiency. Three-month storage at room temperature and lyophilization process does not affect the drug encapsulation efficiency, whereas the size slightly increases to almost 100 nm. The in-vitro drug-release profiles of LNEs suggest that the present formulation can prolong drug release for 48 hours. LNEs can be internalized into tumor cells in vitro and efficiently accumulate in tumor tissues in vivo by passive targeting. Analysis results of in-vitro and in-vivo antitumor activities reveal that doxorubicin-loaded LNE exerts a therapeutic effect similar to that of the commercial Adriamycin. Moreover, the toxicity of doxorubicin, particularly its cardiac toxicity, is reduced. CONCLUSION: The present LNE formulation of doxorubicin can effectively suppress tumor growth and improve the safety of Adriamycin.

[Genetic characteristics of enterovirus 71 VP1 and epidemiology of hand-foot-mouth disease in Xinxiang in 2011].

The study was performed to examine the enterovirus 71(EV71) VP1 genetic feature and the epidemiology of hand-foot-mouth disease (HFMD) in Xinxiang in 2011. Real-time RT-PCR was used for the detection of Pan-enterovirus, Coxsackievirus A 16(CA16) and EV71 from stool specimens of HFMD. The VP1 region was amplified from 10 EV71 positive samples and the products were sequenced. EV71 genotypes were characterized by homology and phylogenetic tree analyses. Additionally, epidemic data of Xinxiang HFMD in 2011 was analyzed. The results revealed that 73% of the specimens from severe cases were determined as EV71 positive, which was significantly higher than CA16-positive ones (19%) (P < 0.01). Ten EV71 strains isolated in Xinxiang belonged to C4a cluster of sub-genotype C4, with 2.8% nucleotide and 0.9% amino acid sequences divergence among them. At position 170 in VP1 gene, an alanine(A) was predominant in 9 isolates, while a valine(V) residue was observed in one isolate. Compared to the representative C4a strains which were closely related to Xinxiang isolates, the amino acid variations of the pre-dominant Xinxiang strains generally occurred at position 292, threonine --> alanine (T --> A). A total of 1118 HFMD cases were reported in Xinxiang in 2011, and 92% of them were younger than 3 years old; the incidence rate peaked in April and December, suggesting that it is very necessary to strengthen HFMD prevention and control even in cold weather.

[Surgical treatment of lymphatic malformation located in head and neck in children].

OBJECTIVE: To evaluate the surgical approaches and therapeutic effect of lymphatic malformations located in head and neck in children. METHODS: Eleven cases of lymphatic malformations in the region of head and neck in children encountered between Jan. 1998 and Dec.2008 in Peking University First Hospital were retrospectively analyzed. Initial diagnosis was made based on the physical examination and then confirmed by MR and Enhanced CT imaging. Surgical therapy was used for patients with lymphatic malformation which exceeds 4 cm. The operative technique was as follows: mass resection and superficial parotidectomy (4 cases), mass resection and total parotidectomy (2 cases), mass resection with neck dissection (2 cases), mass resection with neck dissection and sternotomy (1 case), marginal mandibular branch of facial nerve dissection and mass resection (2 cases). Dissection outside the false capsule was applied during the operation and facial nerve was dissected from bole to terminal arborization. RESULTS: The mass was completely removed in all 11 cases without organ dysfunction and obvious disfigurement. The cure rate was 100%. Three cases suffered from a branch of facial nerve paralysis because of tension and 1 case had a Horner's syndrome after operation. One case needed a blood transfusion (150 ml) during the operation. All cases have been followed up with excellent results from 6 to 121 months, 32 months of the median, no mass recurrence. CONCLUSIONS: Dissection outside the false capsule of mass and dissection of facial nerve were applied in the surgical treatment of huge lymphatic malformations. These methods are effective in the preservation of function and avoidance of abnormality.

[Study on hemorrhage following coblation assisted uvulopalatopharyngoplasty].

OBJECTIVE: To compare the postoperative hemorrhage between standard uvulopalatopharyngoplasty (UPPP) and coblation assisted UPPP, and to evaluate the related risk factors and preventive measures. METHODS: Five hundreds and ninety seven patients with obstructive sleep apnea hypopnea syndrome (OSAHS) underwent UPPP and coblation assisted UPPP between January 1, 1999, and September 30, 2009 were reviewed retrospectively. Two hundred and sixty three patients with coblation assisted UPPP and 334 patients with standard UPPP were treated respectively. Single factor statistic analysis, multiple factors Logistic regress statistic analysis and Wilcoxon test method for related risk factors were applied. RESULTS: A total of 42 patients (7.0%) experienced postoperative bleeding. Among them, 24 patients with coblation assisted UPPP (9.1%) and 18 patients with UPPP (5.4%) had postoperative hemorrhage. Significant difference was not found in the degree of hemorrhage (z = 0.784, P > 0.05), hemorrhage site(χ(2) = 1.387, P > 0.05) and postoperative hemorrhage rates (χ(2) = 3.14, P > 0.05) between the two surgical techniques. Significant difference was found in the interval of hemorrhage after surgery between the two surgical techniques (χ(2) = 9.25, P < 0.01). History of hypertension, smoking, hepatic dysfunction was found to be correlated with the postoperative hemorrhage (Odd-ratio were respectively 7.326, 3.674, 2.707). CONCLUSION: Coblation technique did not significantly increase UPPP postoperative hemorrhage.

[Experiment on inducing human dental pulp stem cells into neural-like cells].

OBJECTIVE: To explore the multi-differentiated capability of human dental pulp stem cells (hDPSCs) obtained by cell-clone culture approach and to determine the appropriate induced medium. METHODS: The cloned isolation and expansion of hDPSCs were preinduced for 24 h, and were subsequently replaced with neural-inductive medium containing certain concentration of dimethylsulfoxide (DMSO), butylated hydroxyanisode (BHA), forskolin, P-mercaptoethanol (p-ME) and hydrocortisone for 4 days. Then induced cells were analyzed by morphological observation, immnocytochemical staining for non-specific esterase (NSE) and glial fibrillary acidic protein (GFAP) expression, RT-PCR for GFAP mRNA. Meanwhile, the uninduced hDPSCs were used as negative control. RESULTS: The morphology of induced cells changed at the initial 12 h, and displayed a typical neuron-like cells at 24 h. There was a gradual increase in the number of these neuronal differentiated cells with continuous induction. Furthermore, immnocytochemical staining showed that the induced cell expressed NSE and GFAP, two marked enzymes of neuron cell. The GFAP mRNA was also detected in induced cells by RT-PCR assay. In contrast, the uninduced cells maintained its original appearance and had no expression on them. CONCLUSION: hDPSCs may possess potential of multiple-differentiation and may differentiate into neuron-like cells on certain inductive condition.

Preparation and evaluation of proliposomes containing clotrimazole.

Clotrimazole (CT)-containing proliposomes were prepared by penetrating an ethanol solution of CT and Egg phosphatidylcholine (PC) into microporous sorbitol particles, followed by vacuum evaporation of the solvent. As a result, CT proliposomes with free-flowing flowability were obtained. On contact with water, the proliposomes were rapidly converted into a liposomal dispersion, in which a certain amount of CT was entrapped by the liposomes. The result in scanning electronic micrograph confirmed the formation of liposomes structures from proliposomes, and the particles revealed round or ellipse. The ratio of drug to total lipid, ratio of PC to cholesterol and ratio of lipid to sorbitol affected the entrapment efficiency (EE%). The EE% of optimized formulation (CT 10 mg, 0.1 g total lipid, PC/CH ratio is 60 : 40 and 1 g sorbitol) in this investigation was 96.2+/-1.5%. The proliposomes system can provide sustaining release in simulated vaginal fluid at 37+/-1 degrees C for 24 h. In-vivo performance of blank proliposomes, a physical mixture of sorbitol and drug, clotrimazole proliposomes and commercial ointment formulation were evaluated using antifungal activity test. At 7 d post-dose, the c.f.u. of C. albicans decreased in proliposomes-treated groups than ointment and the physical mixture (t-Student, p<0.05). The results indicated that CT-containing vaginal proliposomes prolonged drug release and may increase amount of drug retention into the mucosa to result in more antifungal efficacy. In addition, CT-proliposomes did not affect the morphology of vaginal tissues. Therefore, the dosage form might be further developed for safe, convenient, and effective treatment of vaginal candidasis with reduced dosing interval.