Overexpression of Fgf18 in cranial neural crest cells recapitulates Pierre Robin sequence in mice.

The pivotal role of FGF18 in the regulation of craniofacial and skeletal development has been well established. Previous studies have demonstrated that mice with deficiency in Fgf18 exhibit severe craniofacial dysplasia. Recent clinical reports have revealed that the duplication of chromosome 5q32-35.3, which encompasses the Fgf18 gene, can lead to cranial bone dysplasia and congenital craniosynostosis, implicating the consequence of possible overdosed FGF18 signaling. This study aimed to test the effects of augmented FGF18 signaling by specifically overexpressing the Fgf18 gene in cranial neural crest cells using the Wnt1-Cre;pMes-Fgf18 mouse model. The results showed that overexpression of Fgf18 leads to craniofacial abnormalities in mice similar to the Pierre Robin sequence in humans, including abnormal tongue morphology, micrognathia, and cleft palate. Further examination revealed that elevated levels of Fgf18 activated the Akt and Erk signaling pathways, leading to an increase in the proliferation level of tongue tendon cells and alterations in the contraction pattern of the genioglossus muscle. Additionally, we observed that excessive FGF18 signaling contributed to the reduction in the length of Meckel's cartilage and disrupted the development of condylar cartilage, ultimately resulting in mandibular defects. These anomalies involve changes in several downstream signals, including Runx2, p21, Akt, Erk, p38, Wnt, and Ihh. This study highlights the crucial role of maintaining the balance of endogenous FGF18 signaling for proper craniofacial development and offers insights into potential formation mechanisms of the Pierre Robin sequence.

Methotrexate-loaded hyaluronan-modified liposomes integrated into dissolving microneedles for the treatment of psoriasis.

Methotrexate (MTX) is a first-line drug in treating psoriasis because of its strong anti-proliferation and anti-inflammatory effects. However, systemic administration of MTX will lead to many side effects, such as gastrointestinal irritation, liver and kidney toxicity, etc. Herein, we developed liposome-loaded microneedles (MNs) system to improve transdermal efficiency, which was used to overcome the problems of low transdermal efficiency and poor therapeutic effect of traditional transdermal drug delivery methods. Hyaluronic acid (HA) was modified on the surface of MTX-loaded liposomes. The interaction of HA and CD44 could increase the adhesion of HA-MTX-Lipo to HaCaT cells, thereby promoting the apoptosis or death of HaCaT cells. Results indicated HA-MTX-Lipo MNs could inhibit the development of psoriasis and reduce the degree of skin erythema, scaling, and thickening. The mRNA levels of proinflammatory cytokines such as IL-17A, IL-23, and TNF-α were decreased. The epidermal thickness and proliferative cell-associated antigen Ki67 expression were also reduced. Specifically, the expression of mRNA levels of proinflammatory cytokines was down-regulated. The MNs transdermal delivery of HA-modified-MTX liposomes provided a promising method for treating psoriasis.

A novel mutation of gap junction protein ß 1 gene in X-linked Charcot-Marie-Tooth disease.

INTRODUCTION: In this study we report a novel mutation in the gap junction protein beta 1 (GJB1) gene of a Chinese X-linked Charcot-Marie-Tooth disease (CMTX1) family, which has specific electrophysiological characteristics. METHODS: Twenty members in the family were studied by clinical neurological examination and GJB1 gene mutation analysis, and 3 patients were studied electrophysiologically. The proband and his mother also underwent sural nerve biopsy. RESULTS: All patients have the CMT phenotype, except for 2 asymptomatic carriers. Electrophysiological examinations showed non-uniform slowing of motor conduction velocities and partial motor conduction blocks and temporal dispersion. Sural nerve biopsy confirmed a predominantly demyelinating neuropathy, and an Asn2Lys mutation in the amino-terminal domain was found in 9 members of this family, but not in 25 normal controls in the family. CONCLUSIONS: This family represents a novel mutation in the GJB1 form of CMTX1. The mutation in the amino-terminus has an impact on the electrophysiological characteristics of the disease.

High molecular weight hyaluronic acid regulates P. gingivalis-induced inflammation and migration in human gingival fibroblasts via MAPK and NF-κB signaling pathway.

Chronic periodontitis is associated with Porphyromonas gingivalis (P. gingivalis) infection. Hyaluronic Acid (HA), a critical component of the extracellular matrix, exhibits anti-inflammatory and wound-healing properties. This study aimed to investigate the effect of various molecular weights of HA (30, 300 and 1300 kDa) on P. gingivalis-induced inflammatory and wound-healing responses in human gingival fibroblasts (HGFs). Cell cytotoxicity and proliferation were assessed by Lactate dehydrogenase and MTT assays, respectively. An enzyme linked immunosorbent assay was used to detect the levels of interleukin (IL) -1ß, IL-6, IL-8, IL-4 and IL-10. Cell migration was evaluated with a scratch wound healing assay. The expression of nuclear factor kappa B (NF-кB), IкBα, p38 and extracellular signal-regulated kinase (ERK) were analyzed with Western blotting. The results showed that P. gingivalis (1.6 × 106 CFU/mL) and HA (1, 2, 5 and 10 mg/mL) exhibited no toxicity to the HGFs. The 1300 kDa HA inhibited P. gingivalis-induced IL-1ß, IL-6, IL-8, IL-4 and IL-10 production in a dose-dependent manner, while the 30 and 300 kDa HA did not have an effect. Meanwhile, cell migration was significantly promoted by the 30 and 1300 kDa HA. Furthermore, the 1300 kDa HA inhibited NF-κB expression, IκBα degradation and P. gingivalis-induced ERK and P38 activation. Therefore, our study suggests that high molecular weight HA may have beneficial effects on periodontal inflammation and oral wounds.

Transdermal BQ-788/EA@ZnO quantum dots as targeting and smart tyrosinase inhibitors in melanocytes.

Tyrosinase inhibitors could effectively limit the activity of tyrosinase in melanocytes to reduce the excessive synthesis and deposition of melanin. However, low skin permeability and lacking in targeting greatly restricted their application. Herein, ZnO quantum dots were synthesized by gel-sol method and grafted with BQ-788, which have been employed as transdermal and targeting carrier to delivery ellagic acid to melanocytes. Ellagic acid loaded ZnO quantum dots with the size distribution of around 9 nm could targetedly bind to melanocytes and enter the melanocytes by endocytosis within 1 h. The ellagic acid release behavior was controlled by the decreasing of pH via the rapid dissolution of ZnO. When the concentration of BQ-788/EA@ZnO was 12.5 µg/mL, the inhibition rate on tyrosinase activity and melanin deposition were up to 44.23 ±â€¯4.97% and 37.50 ±â€¯5.23%, respectively. In view of their good biocompatibility, they were of great potential in clinically external application for tyrosinase inhibition.

Effects of sodium fluoride on the reproductive development of Bombyx mori.

Bombyx mori was used as a model to evaluate the reproductive toxicity of NaF in insects. Significant reduction in cocoon quality, survival rate, fecundity, and hatchability were observed upon NaF treatment groups. Fluoride determination indicated that F- has a cumulative effect on the gonad of silkworm. High-performance liquid chromatography revealed that the testosterone content of males was decreased in NaF-treated groups, and enzyme-linked immunosorbent assay showed that the estradiol content was decreased in NaF-treated females. Ultrastructural observation of testicles of silkworm larvae revealed mitochondrial turgescence, endoplasmic reticulum destruction, the appearance of vacuoles and lysosomes, and apoptosis and necrosis of cells in NaF-treated groups. Altered tail length and tail DNA content in Comet assays further confirmed DNA damage in NaF-treated larvae. We demonstrated reproductive toxicity of fluoride toward silkworm at physiological and biochemical levels, and the results provide a theoretical basis for revealing the reproductive toxicity of fluoride in terrestrial insects.

Efficient induction of functional ameloblasts from human keratinocyte stem cells.

BACKGROUND: Although adult human tissue-derived epidermal stem cells are capable of differentiating into enamel-secreting ameloblasts and forming teeth with regenerated enamel when recombined with mouse dental mesenchyme that possesses odontogenic potential, the induction rate is relatively low. In addition, whether the regenerated enamel retains a running pattern of prism identical to and acquires mechanical properties comparable with human enamel indeed warrants further study. METHODS: Cultured human keratinocyte stem cells (hKSCs) were treated with fibroblast growth factor 8 (FGF8) and Sonic hedgehog (SHH) for 18 h or 36 h prior to being recombined with E13.5 mouse dental mesenchyme with implantation of FGF8 and SHH-soaked agarose beads into reconstructed chimeric tooth germs. Recombinant tooth germs were subjected to kidney capsule culture in nude mice. Harvested samples at various time points were processed for histological, immunohistochemical, TUNEL, and western blot analysis. Scanning electronic microscopy and a nanoindentation test were further employed to analyze the prism running pattern and mechanical properties of the regenerated enamel. RESULTS: Treatment of hKSCs with both FGF8 and SHH prior to tissue recombination greatly enhanced the rate of tooth-like structure formation to about 70%. FGF8 and SHH dramatically enhanced stemness of cultured hKSCs. Scanning electron microscopic analysis revealed the running pattern of intact prisms of regenerated enamel is similar to that of human enamel. The nanoindentation test indicated that, although much softer than human child and adult mouse enamel, mechanical properties of the regenerated enamel improved as the culture time was extended. CONCLUSIONS: Application of FGF8 and SHH proteins in cultured hKSCs improves stemness but does not facilitate odontogenic fate of hKSCs, resulting in an enhanced efficiency of ameloblastic differentiation of hKSCs and tooth formation in human-mouse chimeric tooth germs.

[Epidemiologic study on STD/HIV infections among tin mining workers in Gejiu, Yunnan province].

OBJECTIVE: To estimate human immunodeficiency virus (HIV)/sexually transmitted diseases (STDs) related knowledge, the prevalence and risk factors for HIV/STDs among tin miners in Gejiu, Yunnan Province. METHODS: A cross-sectional study was conducted in tin miners working at 5 mining regions from March to June 2006 in Gejiu City, Yunnan Province. A total of 1796 miners were enrolled in the study, and a standardized questionnaire was administered probing socio-demographic factors, knowledge of HIV/STDs and sexual behavior. 7 ml-volume venous blood and 15 ml urine specimens were collected for HIV/STDs testing. RESULTS: Twelve participants [0.7%] (12/1760) were identified HIV positive. While, the positive proportions for syphilis, herpes simplex virus type-2 (HSV-2), Neisseria gonorrhea and Chlamydia trachoma were 1.8% (31/1760), 9.6% (169/1760), 0.8% (14/1773), 4.8% (85/1773), respectively. The total prevalence of STDs was 14.9% (264/1776). Factors associated with HIV infection were illegal drug use (adjusted OR = 17.8; 95% CI: 4.0 - 78.8), frequencies of visiting female sex workers in the past twelve months (adjusted OR = 8.7; 95% CI: 1.9 - 39.0), tattoos (adjusted OR = 6.6; 95% CI: 1.8 - 24.0), surgical operation experiences (adjusted OR = 6.0; 95% CI: 1.6 - 22.5) and toothbrush sharing (adjusted OR = 5.6; 95% CI: 1.0 - 31.3). Factors associated with STDs infection were race (adjusted OR = 2.0; 95% CI: 1.5 - 2.7), age (adjusted OR = 1.7; 95% CI: 1.0 - 2.9), illegal drug use (adjusted OR = 2.3; 95% CI: 1.0 - 5.2), living with spouses (adjusted OR = 1.4; 95% CI: 1.1 - 1.9) and visiting female sex workers (adjusted OR = 1.9; 95% CI: 1.4 - 2.6). Only 4.1% (49/1201) of respondents aware the correct transmission routes of HIV. 21.6% (339/1569) of the miners who had previous sexual experience self-reported visiting female sex workers. CONCLUSION: HIV and other STDs are highly prevalent amongst miners in this region. In addition, the awareness of HIV/STDs is low and multiple routes of HIV transmission, such as heterosexual intercourse and IDU, were also observed in these study subject. HIV/STDs interventions should therefore be reinforced among the miners in this region.

[Study of three-dimensional anatomical relationship between infrazygomatic crest and maxillary sinus for mini-screw insertion].

PURPOSE: To measure the thickness of the infrazygomatic(IZ) crest above the maxillary first molar and analyze its relationship with maxillary sinus as to derive clinical implications and provide guidance for placements of orthodontic mini-screw. METHODS: Thirty seven cone-beam CT(CBCT) images of infrazygomatic crest sites were collected from data of 19 adult inter-treatment orthodontic patients. Based on the relationship of maxillary sinus floor and mesiobuccal root of the maxillary first molar, the subjects were divided into high sinus floor group (n=13) and low sinus floor group (n=24). Both lengths of mesial and distal buccal root of the maxillary first molar were measured as a reference for mini-screw positioning. Statistical analysis of the data was conducted using SPSS 20.0 software package. RESULTS: IZ crest thickness above the maxillary first molar in high sinus group was larger than that in low sinus group, and the thickness decreased as the measurement moved towards the skull base in both groups. Lengths of mesial and distal buccal root of the maxillary first molar were (12.62±0.55)mm and (12.30±0.48)mm respectively in male patients alongside (11.53±0.24)mm and (11.15±0.26)mm respectively in female patients. CONCLUSIONS: Our data indicated pre-insertion evaluation of the appropriate level of maxillary sinus floor should be performed before placement of mini-screw using CBCT. In order to avoid injury to the mesiobuccal root of the maxillary first molar and enhance the stability of inserted miniscrew, positions for miniscrew implant insertion should be over 13.62 mm in Chinese male patients and 12.53 mm in female patients at the vertical level from buccal CEJ.

[A cross-sectional survey on human immunodeficiency virus infection in a former commercial blood donating community, Shanxi Province].

OBJECTIVE: To determine the sero-prevalence of and the risk factors for human immunodeficiency virus infection among residents in a former commercial blood donating community, Shanxi Province. METHODS: A community-based cross-sectional survey was conducted among all residents aged 18 - 64 years in 4 villages in a rural county of Shanxi Province. A standardized structured questionnaire was administered to collect socio-demographic, medical and risk behavioral information. Venous blood was collected for HIV antibody testing. Generalized estimating equation with logistic regression was used to analyze the risk factors for HIV infection. Population attributable fraction was used to evaluate the role of public health in the risk factors for the incidence of HIV. RESULTS: A total of 3062 villagers participated in the study and provided their blood samples for HIV antibody testing. 29.5% of the participants had a history of selling blood and/or plasma. Among the subgroup of former blood donors (FBDs), 71.9% sold whole blood only, 4.1% sold plasma only and 24.0% sold both. Forty HIV infected persons were identified in the four villages. HIV prevalence was 1.3%, overall, 4.1% in FBDs and 0.1% in non-FBDs in the community. Amongst the subgroup of FBDs, HIV prevalence was 1.5% for whole blood only, 21.6% for plasma only and 8.8% for both. Univariate analysis showed that village of residence, education, marital status, gender, plasma/blood donation were statistically associated with HIV seropositivity in the villagers. Other variables, such as gender, age, occupation, number of sexual partners, drug use, histories of tooth extraction, acupuncture, surgical operation and medical injection were not found to be statistically associated with HIV seropositivity. Multivariate analysis found that selling whole blood only (adjusted OR = 13.25, 95% CI: 3.02 - 58.25) and selling plasma or both (adjusted OR = 87.85, 95% CI: 22.22 - 347.27) were significantly associated with HIV seropositivity, and the additive population attributable fraction (PAF) was 89.8%. Villagers in village 03 were more likely to be HIV positive than those in village 04 (adjusted OR = 12.26, 95% CI: 1.46 - 103.00). CONCLUSION: At present, HIV epidemic in the former commercial blood donating community has a low prevalence in Shanxi Province and the majority of HIV infections should be related to the blood and plasma donation. This study reveals the existence of HIV second-generation transmission from the infected blood donors to the general population. However, HIV had not been evidently extended from the blood/plasma donors to general population.

Underwater self-cleaning scaly fabric membrane for oily water separation.

Oily wastewater is always a threat to biological and human safety, and it is a worldwide challenge to solve the problem of disposing of it. The development of interface science brings hope of solving this serious problem, however. Inspired by the capacity for capturing water of natural fabrics and by the underwater superoleophobic self-cleaning property of fish scales, a strategy is proposed to design and fabricate micro/nanoscale hierarchical-structured fabric membranes with superhydrophilicity and underwater superoleophobicity, by coating scaly titanium oxide nanostructures onto fabric microstructures, which can separate oil/water mixtures efficiently. The microstructures of the fabrics are beneficial for achieving high water-holding capacity of the membranes. More importantly, the special scaly titanium oxide nanostructures are critical for achieving the desired superwetting property toward water of the membranes, which means that air bubbles cannot exist on them in water and there is ultralow underwater-oil adhesion. The cooperative effects of the microscale and nanoscale structures result in the formation of a stable oil/water/solid triphase interface with a robust underwater superoleophobic self-cleaning property. Furthermore, the fabrics are common, commercially cheap, and environmentally friendly materials with flexible but robust mechanical properties, which make the fabric membranes a good candidate for oil/water separation even under strong water flow. This work would also be helpful for developing new underwater superoleophobic self-cleaning materials and related devices.

The effect of thick fibers and large pores of electrospun poly(ε-caprolactone) vascular grafts on macrophage polarization and arterial regeneration.

The vascular grafts prepared by electrospinning often have relatively small pores, which limit cell infiltration into the grafts and hinder the regeneration and remodeling of the grafts into neoarteries. To overcome this problem, macroporous electrospun polycaprolactone (PCL) scaffolds with thicker fibers (5-6 µm) and larger pores (∼30 µm) were fabricated in the present study. In vitro cell culture indicated that macrophages cultured on thicker-fiber scaffolds tended to polarize into the immunomodulatory and tissue remodeling (M2) phenotype, while those cultured on thinner-fiber scaffolds expressed proinflammatory (M1) phenotype. In vivo implantation by replacing rat abdominal aorta was performed and followed up for 7, 14, 28 and 100 d. The results demonstrated that the macroporous grafts markedly enhanced cell infiltration and extracellular matrix (ECM) secretion. All grafts showed satisfactory patency for up to 100 days. At day 100, the endothelium coverage was complete, and the regenerated smooth muscle layer was correctly organized with abundant ECM similar to those in the native arteries. More importantly, the regenerated arteries demonstrated contractile response to adrenaline and acetylcholine-induced relaxation. Analysis of the cellularization process revealed that the thicker-fiber scaffolds induced a large number of M2 macrophages to infiltrate into the graft wall. These macrophages further promoted cellular infiltration and vascularization. In conclusion, the present study confirmed that the scaffold structure can regulate macrophage phenotype. Our thicker-fiber electrospun PCL vascular grafts could enhance the vascular regeneration and remodeling process by mediating macrophage polarization into M2 phenotype, suggesting that our constructs may be a promising cell-free vascular graft candidate and are worthy for further in vivo evaluation.

Starch derivative-based superabsorbent with integration of water-retaining and controlled-release fertilizers.

Phosphate rock (PHR), a traditional fertilizer, is abundant, but is hard to be utilized by plants. To improve the utilization of PHR, and to integrate water-retaining and controlled-release fertilizers, an agricultural superabsorbent polymer based on sulfonated corn starch/poly (acrylic acid) embedding phosphate rock (SCS/PAA/PHR) was prepared. PHR can be suspended and well-dispersed in SCS/PAA by sulfonated corn starch (SCS). PHR and KOH were mixed in acrylic acid solution to provide phosphorus (P) and potassium (K) nutrients, respectively. Impacts on water absorption capacity of the superabsorbent were investigated. The maximum swelling capacity in distilled water or 0.9 wt.% (weight percent) NaCl solution reached 498 g g(-1) and 65 g g(-1) (water/prepared dry superabsorbent) respectively. Moreover, release behaviours of P and K in SCS/PAA/PHR were also investigated. The results showed that SCS/PAA/PHR possessed excellent sustained-release property of plant nutrient, and the SCS/PAA could improve the P release greatly. Besides, the XPS analysis was employed to study the relationship between PHR and superabsorbent polymer.

Sugarcane bagasse derivative-based superabsorbent containing phosphate rock with water-fertilizer integration.

To improve the water-fertilizer utilization ratio and mitigate the environmental contamination, an eco-friendly superabsorbent polymer (SPA), modified sugarcane bagasse/poly (acrylic acid) embedding phosphate rock (MSB/PAA/PHR), was prepared. Ammonia, phosphate rock (PHR) and KOH were admixed in the presence of acrylic acid to provide nitrogen (N), phosphorus (P) and potassium (K) nutrients, respectively. Impacts on water absorption capacity of the superabsorbent polymer (SAP) were investigated. The maximum swelling capacity in distilled water and 0.9 wt.% (weight percent) NaCl solution reached 414 gg(-1) and 55 gg(-1) (water/prepared SAP), respectively. The available NPK contents of the combination system were 15.13 mgg(-1), 6.93 mgg(-1) and 52.05 mgg(-1), respectively. Moreover, the release behaviors of NPK in the MSB/PAA/PHR were also studied. The results showed that the MSB/PAA/PHR has outstanding sustained-release plant nutrients property.

[Maxillary protraction with and without maxillary expansion on unilateral cleft lip and palate model: a finite element analysis].

PURPOSE: The aim of this finite element study was to investigate biomechanical effects of maxillary protraction with and without maxillary expansion on unilateral cleft lip and palate(UCLP)model before and after alveolar bone graft (ABG). METHODS: Orthopedic force (30° downward and forward to the occlusal plane, 5N/side) with expansion and without expansion were loaded on models before ABG, and after ABG respectively. Initial displacement and sutural stress were analyzed by using ANSYS12.0 software. RESULTS: The amount of displacement at the cleft side was larger than that at the non-cleft side before ABG, a more symmetrical pattern was obtained after ABG. Regardless of ABG, maxillary protraction with maxillary expansion exhibited more evident forward displacement of maxilla. The sutural stress values after maxillary protraction with maxillary expansion were significantly higher than without expansion. CONCLUSIONS: Analysis of these results showed that maxillary protraction with expansion could presumably promote the orthopedic effects of the facemask on UCLP patients, and more uniform force would obtain after ABG.

Endothelialization and patency of RGD-functionalized vascular grafts in a rabbit carotid artery model.

To address the growing demand of small-diameter vascular grafts for cardiovascular disease, it is necessary to develop substitutes with bio-functionalities, such as anticoagulation, rapid endothelialization, and smooth muscle regeneration. In this study, the small-diameter tubular grafts (2.2 mm) were fabricated by electrospinning of biodegradable polymer polycaprolactone (PCL) followed by functional surface coating with an arginine-glycine-aspartic acid (RGD)-containing molecule. The healing characteristics of the grafts were evaluated by implanting them in rabbit carotid arteries for 2 and 4 weeks. Results showed that at both time points, all 10 of the RGD-modified PCL grafts (PCL-RGD) were patent, whereas 4 of the 10 non-modified PCL grafts were occluded due to thrombus formation. Scanning electron microscopy (SEM) data showed abundant platelets adhering on the surface of the midportion of the PCL grafts. In contrast, only few platelets were observed on the PCL-RGD surface, suggesting that RGD modification significantly improved the hemocompatibility of the PCL grafts. Histological analysis demonstrated enhanced cell infiltration and homogeneous distribution within the PCL-RGD grafts in comparison with the PCL grafts. Furthermore, immunofluorescence staining also showed a 3-fold increase of endothelial coverage of the PCL-RGD grafts than that of PCL grafts at those two time points. After 4-week implantation, 65.3 ± 7.6% of the surface area of the PCL-RGD grafts was covered by smooth muscle cell layer, which is almost 23% more than that on the PCL grafts. The present study indicates that RGD-modified PCL grafts exhibit an improved remodeling and integration capability in revascularization.

Development of glycyrrhetinic acid-modified stealth cationic liposomes for gene delivery.

The glycyrrhetinic acid-modified stealth cationic liposomes (GA-PEG-CLs) loaded with pDNA (GA-PEG-CLPs) were developed and found to transfect human hepatocellular carcinoma cell line HepG2 with high efficiency. GA-PEG-CLs were comprised of DOTAP, cholesterol (Chol) and glycyrrhetinic acid-polyethyleneglycol-cholesterol conjugate (GA-PEG-Chol). Agarose gel electrophoresis revealed that 5% GA-PEG-CLs constituted by DOTAP/Chol/GA-PEG-Chol at molar ratio of 50:45:5 could completely entrap pDNA at a lower liposomes/pDNA weight ratios of 4:1 (N/P ratio: 1.14). Compared to ordinary cationic liposomes (CLs), steric cationic liposomes (PEG-CLs) and 1% GA-PEG-CLs made from DOTAP/Chol/MPEG2000-Chol/GA-PEG-Chol at molar ratio of 50:45:4:1, 5% GA-PEG-CLs were found to possess the highest transfection efficiency as gene vectors in serum-free or serum-containing medium in PKCalpha over-expressed HepG2 cells but no significance difference in human embryonic kidney cell line HEK 293. Additionally, 5% GA-PEG-CLs have the lowest cytotoxicity on human normal hepatocyte cell line L02. The competitive inhibition experiments mediated by GA were carried out in HepG2 cells, which demonstrated that GA-PEG-CLs could deliver selectively pDNA to hepatoma cells by the targeting moiety GA. In conclusion, GA-PEG-CLs containing 5% GA-PEG-Chol might be one of the most potential gene vectors as hepatoma targeting therapy.