Effects of toothpaste containing inactivated Lacticaseibacillus paracasei Probio-01 on plaque-induced gingivitis and dental plaque microbiota.

Plaque-induced gingivitis is an inflammatory response in gingival tissues resulting from bacterial plaque accumulation at the gingival margin. Postbiotics can promote the proliferation of beneficial bacteria and optimise the state of microbiota in the oral cavity. In this study, we investigated the effect of inactivated Lacticaseibacillus paracasei Probio-01 on plaque-induced gingivitis and the dental plaque microbiota. A total of 32 healthy gingival participants (Group N, using blank toothpaste for 3 months) and 60 patients with plaque-induced gingivitis (30 in Group F, using inactivated Probio-01 toothpaste for 3 months, and 30 in Group B, using blank toothpaste for 3 months, respectively) were recruited. Clinical indices, which included bleeding on probing (BOP), gingival index (GI), and plaque index (PI), were used to assess the severity of gingivitis. Furthermore, 16SrDNA amplicon sequencing was used to explore changes in the gingival state and dental plaque microbiota in patients with plaque-induced gingivitis. The results showed that inactivated Probio-01 significantly reduced clinical indices of gingivitis, including BOP, GI, and PI, in participants with plaque-induced gingivitis and effectively relieved gingival inflammation, compared with that observed in the control group (group B). Inactivated Probio-01 did not significantly influence the diversity of dental plaque microbiota, but increased the relative abundance of dental plaque core bacteria, such as Leptotrichia and Fusobacterium (P < 0.05). Strong correlations were observed between the indices and abundance of dental plaque microbiota. Overall, the inactivated Probio-01 significantly reduced the clinical indices of gingivitis and effectively improved gingival inflammation in patients with plaque-induced gingivitis. The activity of inactivated Probio-01 against plaque-induced gingivitis was possibly mediated by its ability to regulate the dental plaque microbiota, as indicated by the close correlation between the plaque microbiota and clinical indices of gingivitis.

Dental visit experience and dental care barriers among Hong Kong preschool children with special education needs.

BACKGROUND: Children with special education needs (SEN) require additional supports in education settings, as well as extra supports in healthcare services. AIM: To investigate dental visit experience and dental care barriers among Hong Kong preschoolers with SEN. DESIGN: This was designed as a cross-sectional study. A structured questionnaire was employed to investigate dental attendance and dental care barriers among Hong Kong preschoolers with SEN. Binary logistic regression and multi-factor ANOVA were performed to identify factors associated with children's dental visit experience. RESULTS: A total of 383 children were included in the final analysis. The average age of the recruited children was 3.87 ± 0.95 years. 72.5% parents reported that dental visits were a challenging task for their children. Among children with caries, 70.7% had never visited a dentist. Regression models indicated that (a) children over 3 years or children whose parents received tertiary education were more likely to visit a dentist; (b) the existence of dental care barriers was associated with children's primary diagnosis, practical skills, conceptual skills, and parents' education attainment; and (c) children with low conceptual skills (95%CI:0.71 to 2.40, P < .001), or children from low-income families (95%CI:1.21 to -0.06, P = .031) had more dental care barriers than their counterparts. CONCLUSION: Unmet dental treatment needs and dental care barriers existed among pre-schoolers with SEN. Dental care barriers were associated with children's developmental profile, parents' education attainment, and household income.

Toothbrush deterioration and parents' suggestions to improve the design of toothbrushes used by children with special care needs.

BACKGROUND: Biting objects was a parafunctional oral habit among children with special care needs. Chewing or biting toothbrushes could expedite the process of toothbrush wear. However, few studies evaluated the deterioration levels of toothbrushes used by children with special needs. This study aimed to assess the deterioration level of toothbrushes used by children with special care needs, and collect parents' feedbacks to improve the design of children's toothbrushes. METHODS: The cross-sectional study recruited 277 children who had special care needs. Children's toothbrushing behaviors, background information, and parents' comments on toothbrushes were obtained. Toothbrush deterioration was assessed by bristle wear and bite mark scores. Higher scores indicated severe deterioration. RESULTS: Three hundred twenty-one toothbrushes were collected. Children who used 2 to 6 toothbrushes in a 3-month period showed higher toothbrush deterioration scores than children who used a single toothbrush. Over 40% children's toothbrushes presented excessive wear. Excessive wear was associated with social skills and parents' education background. Distinct bite marks tended to exist on toothbrushes which had been used by children who showed challenging behaviors during toothbrushing (OR = 1.96, 95%CI1.15-3.32, p < 0.05). Approximately 27% parents reported that children's toothbrushes should be modified. Parents recommended that the size of toothbrush heads, the angle of handles, and the texture/length/distribution/diameter of bristles should be adjusted. Besides, ideal toothbrushes should be able to provide verbal or visual instructions to children, motivate children to brush teeth, simplify toothbrushing procedure, and protect children who had toothbrush-biting habits. CONCLUSIONS: Excessive wear and distinct bite marks can be found on toothbrushes that had been used by children with special care needs. Toothbrush deterioration was associated with children's social skills, toothbrushing behaviors, and parents' educational attainment. The commercially available toothbrushes should be modified to meet the additional needs of young children.

Oral health and associated factors among preschool children with special healthcare needs.

OBJECTIVE: To investigate the oral health status and associated factors among preschool children with special healthcare needs. SUBJECTS AND METHODS: This was designed as a cross-sectional study. Dental examination was provided to 383 children at Special Child Care Centres. Covariates of interest included children's age, developmental profile, oral health-related behaviours and family social-economic status. The association between children's dental caries experiences, dental plaque deposition and gingival health status were analysed by negative binomial regression, multiple factor ANOVA and binary logistic regression when appropriate. RESULTS: The proportion of children with caries, gingival inflammation and plaque were 30.3%, 89.6% and 95.0%, respectively. Final regression models indicated that (a) tooth decay was associated with dental visit experiences, night bottle-feeding habit, fluoridated toothpaste and monthly income; (b) plaque deposition was associated with intellectual functioning, practical skills, night bottle-feeding habit, use of toothpaste, initiation of toothbrushing habit, toothbrushing duration and additional cleaning habit; (c) gingival status was associated with children's intellectual functioning and toothbrushing frequency. CONCLUSIONS: Poor oral hygiene status existed among preschool children with special healthcare needs. Children's oral health status was associated with their developmental profile, oral health-related behaviours and social-economic status.

The Impact of Adaptive Functioning and Oral Hygiene Practices on Observed Tooth-Brushing Performance Among Preschool Children with Special Health Care Needs.

OBJECTIVES: To investigate the impact of adaptive functioning and oral hygiene practices on tooth-brushing performance among preschool children with special health care needs (SHCN). METHODS: A cross-sectional study was conducted in Special Child Care Centers. Children's tooth-brushing performance was assessed by a standardized 13-step pro forma. Information regarding children's socio-economic status, adaptive skills, and oral hygiene practices were collected. Bivariate analysis and ANCOVA were used to explore the potential factors which might be associated with children's tooth-brushing performance. RESULTS: The tooth-brushing assessment was provided to 379 children with SHCN. Approximately 3% of the recruited children performed the whole tooth-brushing procedure independently. The number of tooth-brushing steps practiced by those children was 4.47 ± 3.56. Children who had established tooth-brushing habit before age one practiced more tooth-brushing steps than children who brushed their teeth after age one (p = 0.029). When children's age, gender, and socio-economic status were adjusted, children who had established regular tooth-brushing habit or children who had high levels of adaptive skills showed better tooth-brushing performance than their peers. Children who used gauze, cotton swab, or dental floss to clean their teeth practiced fewer key tooth-brushing steps than their peers who had never used additional cleaning approaches (p = 0.038). CONCLUSIONS FOR PRACTICE: Children's tooth-brushing performance was associated with adaptive skills and oral hygiene practices. Tooth-brushing training should be provided to children with SHCN in early childhood. For children who had limitations in adaptive functioning, parental assistance or supervision is recommended to guarantee the efficacy and safety of daily tooth brushing.

A systematic review and meta-analysis of dental plaque control among children and adolescents with intellectual disabilities.

BACKGROUND: People with intellectual disabilities (ID) typically have poorer oral health. Oral diseases have a microbiological origin, and thus, dental plaque control (DPC) is key to safeguard their oral health. METHODS: A structured search strategy was applied to screen oral health promotion initiatives for DPC among children and adolescents with ID following PRISMA statements. Qualitative and quantitative analyses were performed. RESULTS: From 1,455 citations, 22 studies informed this review. Seven studies related to mechanical interventions and 13 to chemical interventions. Qualitative analyses found conventional toothbrushes were less effective than modified toothbrushes but quantitative analyses were conflicting. Of the nine studies that compared chlorhexidine to placebos, seven reported chlorhexidine was more effective than placebos. Meta-analysis favoured chlorhexidine to placebos (p < 0.0001, 95% CI 0.34-0.63). CONCLUSIONS: Both mechanical and chemical interventions are effective for DPC. It is unclear whether they are alternative means for DPC or whether they are best employed together.

Effectiveness of a visual-verbal integration model in training parents and their preschool children with intellectual and developmental disabilities to dispense a pea-sized amount of fluoridated toothpaste.

BACKGROUND: A visual-verbal integration model (VVIM) was used to train parents and their children with intellectual and developmental disabilities (IDD) to dispense a pea-sized amount of fluoridated toothpaste, aiming to balance the occurrence of dental caries and fluorosis. METHOD: Participants were 370 pairs of IDD preschool children and their parents. A two-phase pre-post-intervention study was performed. The weight of toothpaste was calculated by a portable electronic compact balance. RESULTS: A quarter of parents and <20% children dispensed the appropriate amount at baseline. Almost 85% parents and 50% children achieved the pea-sized amount after intervention. Half of the children continued to use the recommended toothpaste amount after 6 months. Children with higher intellectual functioning and conceptual skills were more likely to dispense the pea-sized amount of toothpaste. CONCLUSIONS: Visual-verbal integration model training had a desired impact on parents' toothpaste-dispensing performance. Children with higher adaptive skills may benefit from VVIM training.

Oral health status of children and adolescents with intellectual disabilities: a systematic review and meta-analysis.

AIM: To compare the oral health status of children and adolescents affected by intellectual disabilities with their unaffected counterparts. METHOD: Citations published in English were searched from electronic databases (PubMed, Embase, Web of Science, and Scopus) from their start dates to March 2017. The whole process was conducted following PRISMA (Preferred Reporting Items for Systematic reviews and Meta-Analyses) guidelines. The PICO (population, intervention/interest, comparator, outcome) principle was used to formulate the topic. Studies were synthesized through qualitative summary or, whenever possible, meta-analysis. RESULTS: The initial search yielded 2393 records. Thirty-nine studies from 22 countries were identified for qualitative analysis; 26 studies were eligible for meta-analysis. Participants with intellectual disabilities had higher levels of dental plaque, worse gingival status, fewer decayed and filled permanent teeth, and similar caries experience between males and females. These findings were supported by both qualitative and quantitative analysis. Various patterns of caries experiences were indicated by qualitative analysis, but it was not substantiated by meta-analysis. INTERPRETATION: There is increasing worldwide interest in oral health status of children with intellectual disabilities. Differences in dental plaque deposition, gingival inflammation, and the number of decayed and filled permanent teeth were investigated between children and adolescents with and without intellectual disabilities. Evidence remains elusive about the pattern of caries experience among those children.

Management of oblique root fracture in the middle third of two maxillary immature central incisors with severe caries in mixed dentitions: a case report.

Mid-root fractures are rare injuries in young permanent teeth and tend to have poor prognoses. This study presents a case of oblique root fracture of both maxillary immature central incisors in the middle third accompanied by delayed dental visit and severe caries of all primary teeth. After restoring all the primary and permanent teeth that needed stabilization, the coronal fragments were repositioned and stabilized with a flexible splint consisting of orthodontic wire and composite resin. A comprehensive and sequential dental treatment for other oral diseases and oral hygiene instructions were provided. A 16-month follow-up revealed that the two injured young permanent incisors were healed, surrounded by hard tissues and continued to grow both in length of the root and thickness of the root canal wall, with significant improvement in oral hygiene. Based on the outcome of this case, initial stabilization without endodontic therapy could be considered a successful treatment modality for young permanent teeth with oblique root fracture due to the growth of fractured teeth with vital pulp and the maintenance of natural dentition.

Social Story Intervention for Training Expected Behaviors among Preschool Children: A Systematic Review and Meta-Analysis.

Promoting appropriate behaviors in early childhood is crucial for children's future development. This systematic review aimed to explore the efficacy of social story (SS) intervention in teaching expected behaviors among preschool children. A structured search strategy was applied to five online electronic databases. The references were systematically screened in accordance with the PRISMA statements. Randomized or non-randomized controlled studies, as well as single-subject studies, in which SSs served as a behavioral training approach for children aged 2 to 6 years were included. Information related to study design, characteristics of the participants, target behaviors, and implementation of SS intervention was extracted. A meta-analysis was performed using the random-effects model, where similar outcomes were evaluated by similar intervention across multiple studies. Twenty-one studies were identified for qualitative analysis, while two studies formed the basis of the meta-analysis. SS interventions were employed to teach a variety of behaviors among typically developing children as well as those with various disabilities, such as autism, developmental delay, hearing impairments, attention deficit hyperactivity disorder, or other disabilities. The target behaviors included oral health practices, peer interaction, staying on-task, self-regulation, sleep habits, and controlling aggressive behavior during group activities. The SSs were used either alone or combined with other strategies, such as positive reinforcement, music therapy, role play, group discussion, video self-modeling, immediate practices, or additional audio commentary. Most studies reported improvements in appropriate behaviors and/or reductions in unfavorable behaviors. The meta-analysis indicated that children practiced more toothbrushing steps when using SS interventions compared to conventional oral health instruction (Z = 3.60, MD = 0.66, 95%CI 0.30 to 1.02, p < 0.001). SS interventions have the potential to teach target behaviors, particularly toothbrushing behaviors, among preschool children. More well-designed randomized controlled trials are warranted to determine the efficacy of SS interventions among children with various developmental profiles.

[Dental treatment under general anesthesia among pediatric patients in 120 consecutive cases: a retrospective study].

PURPOSE： To investigate the clinical features of children who received treatment under dental general anesthesia (DGA). METHODS: The clinical records of dental patients below 18 years old who were treated under DGA at the Department of Pediatric Dentistry, Affiliated Dental Hospital of Kunming Medical University during June 2017 to November 2019 were obtained, including the baseline information, causes for DGA, anesthesia methods, intubation methods, treatment items, treatment time and follow-up visits. SPSS 26.0 software package was used to analyze the data. RESULTS: A total of 120 patients were included, 58.3% were males, and children aged 3 to 6 years showed the highest demand for DGA (85.0%). Fear of dental treatment, ineffective non-drug behavior management was the main causes for DGA in young children, while the most common causes for children over 6 years old to choose DGA were mental retardation (38.9%) and patients' needs(38.9%). The average number of teeth treated was (15.16±3.42) for each child, and the average time for treating one tooth was 12.26 min. Restoration, root canal treatment and primary teeth pre-forming crown(including anterior preformed resin transparent crown and posterior preformed metal crown) were the main treatment items. At 1-week follow-up visits, 98.3% of children had no discomfort. During 2017 to 2019, there was an increasing tendency in the number of patients who chose DGA in the authors' institute. CONCLUSIONS： The dental issues of children with fear of dental treatment, ineffectiveor non-drug behavior management or mental retardation can be treated under DGA conveniently, safely and efficiently. The acceptance rate of DGA among pediatric patients is on the rise. DGA training programs and related support projects are needed to meet the treatment demands among patients in less developed areas.

Impact of Antiretroviral Therapy on Oral Health among Children Living with HIV: A Systematic Review and Meta-Analysis.

Oral health is an integral component of general health and well-being but might be undermined among children living with HIV (CLWH) due to the condition itself or the antiretroviral therapy (ART) received. This review summarises the current evidence and compares the oral health status of the CLWH who were treatment-naïve with those undergoing different ART medications. Fourteen studies were included in the final qualitative and quantitative analyses. This review identified no significant difference in the prevalence of caries, periodontal conditions, and tooth development between both groups. Orofacial opportunistic infections were more prevalent in the CLWH without ART. Children undergoing ART with a duration longer than 3 years had a significantly lower prevalence of oral candidiasis and CD4+ T-cell counts. However, due to the insufficient number of well-administered case-control studies with adequate sample size, the quality of the evidence in all outcomes was of very low certainty.

Oral Health Status of Children and Adolescents Living with HIV Undergoing Antiretroviral Therapy: A Systematic Review and Meta-Analysis.

Antiretroviral therapy (ART) increases the survival of HIV-infected children, but might also bring in oral health-related side effects and increase their risks of oral diseases. The review compared the oral health status of children living with HIV (CLWH) undergoing ART with healthy controls. Dual independent screening and study selection from four electronic databases and manual searches, data extraction, risk of bias assessment, and quality-of-evidence evaluation with Grading of Recommendations Assessment Development and Evaluation were performed. Twelve studies were included in qualitative and quantitative analysis. CLWH taking ART had a significantly higher prevalence of periodontal diseases (OR = 3.11, 95% CI 1.62-5.97), mucosal hyperpigmentation (OR = 20.35, 95% CI 3.86-107.39), and orofacial-related opportunistic infections than healthy controls. No significant differences regarding caries prevalence and tooth development were identified. Those with CD4+ T-cell counts below 250 cells/mm3 were more likely to manifest opportunistic infections, while medication duration had minimal influence on the prevalence of orofacial opportunistic infections. The current findings did not identify HIV and antiretroviral status as predisposing factors to dental caries, but affirmed the associated increased risk of periodontal diseases, mucosal hyperpigmentation and candidiasis.

Dental Caries Status among Yi Preschool Children in Yunnan Province, China: A Cross-Sectional Study.

BACKGROUND: The Yi ethnic group is the sixth largest minority in China. The aim of this study was to investigate dental caries status among Yi preschool children in the Yunnan province, China. METHOD: This cross-sectional study invited 5-year-old Yi children using multistage cluster sampling. Two trained and calibrated dentists examined the children in kindergartens. The children's dental caries experience was assessed using the decayed, missing, and filled teeth (dmft) index. Visual plaque on an indexed tooth of each six sextants was recorded and the oral hygiene status was assessed using the Visible Plaque Index. Parents completed questionnaires that surveyed their educational attainment. The questionnaires also collected information about the children's demographics, snacking habits, oral-health-related behaviors and dental visit experiences. The associations between caries experiences and potential factors were analyzed using zero-inflated negative binomial regression. RESULTS: All 452 invited children (249 boys, 55%) participated in this study with a response rate of 100%. Dental caries prevalence rate was 83%. The mean dmft score and decayed teeth score were 5.2 ± 4.4 and 5.1 ± 4.4, respectively. Almost half of the children (n = 211, 47%) had visible plaque on four or more of the six sextants. Most (n = 366, 81%) of the children had not visited a dentist in the prior 12 months. Regression analysis found the children's caries experience was associated with their dental visit experience. CONCLUSION: The great majority of the Yi preschool children experienced dental caries and almost all of the cavities were not restored. Their oral hygiene was poor and visible plaque was commonly found on their teeth.

Oral Health Status of 12-Year-Old Hani Children in the Yunnan Province of China: A Cross-Sectional Study.

BACKGROUND: The aim of this cross-sectional study was to investigate the oral health status of Hani 12-year-old children in Yunnan, a province in Southwest China. METHOD: This study employed a multistage sampling method to recruit children from local primary schools. Two calibrated dentists examined the status of dental caries, gingival bleeding and dental fluorosis by adopting the diagnosis criteria recommended by the World Health Organization. A self-administrated questionnaire was distributed. The chi-square test and multivariate logistic regression were conducted. RESULTS: This study invited 480 Hani children, and recruited 413 children (52% boys) (response rate: 86%). The dental caries prevalence was 52%, and the caries experience associated with the mean (standard deviation) decayed, missing and filled teeth (DMFT) score was 1.10 (1.46). Gingival bleeding was diagnosed in 252 children (61%), and dental fluorosis was found in two children (0.5%). The results of the regression model indicated the prevalence of dental caries were associated with sugary snacking habits (p = 0.002). The prevalence of gingival bleeding was related to the mother's education level as well as the child's monthly pocket money (p < 0.05). CONCLUSION: Dental caries and gingival bleeding were prevalent among 12-year-old Hani children in the Yunnan province in China. Dental fluorosis was uncommon.

Social story-based oral health promotion for preschool children with special healthcare needs: A 24-month randomized controlled trial.

OBJECTIVES: This study aimed to investigate the efficacy of the social story-based intervention in promoting the oral health status among preschool children with special healthcare needs. METHODS: The 24-month randomized controlled trial recruited 306 children from centres that provided educational training for 2- to 6-year-old children who had special healthcare needs. The participants received standardized toothbrushing training and oral health education materials. The education materials for children in the test group were validated social stories, while materials for children in the control group were standard leaflets. Those materials were introduced in each training session and used at home by parents. Children's oral health status and oral health-related behaviours were assessed before and after interventions. Data were analysed via generalized estimating equations, negative binomial regression and binary logistic regression. RESULTS: There was no significant difference in caries experience among children in both groups after 24 months. However, children in the test group showed better oral hygiene status and better toothbrushing performance, and they were more likely to visit a dentist (OR = 2.00, 95% CI:1.15,3.49) than their peers in the control group. Caries progression was more likely to be observed among children who had low social skills or whose parents were not working full-time. Children from high-income families showed better oral hygiene status than their peers (Estimate = -0.11, 95%CI: -0.20,-0.01). CONCLUSION: Social stories could be used to improve oral health-related behaviours among young children with SHCN. The efficacy of the intervention might be associated with children's adaptive skills and family socio-demographic status.

Efficacy of Social Story Intervention in Training Toothbrushing Skills Among Special-Care Children With and Without Autism.

Toothbrushing is a routine practice for oral hygiene maintenance. It is also a challenging task for young children with special care needs. In this study, social stories were employed to teach toothbrushing skills for preschool children with special needs. Those children were recruited from the Special Child Care Centers, including 87 children with autism and 94 children without autism. A validated toothbrushing social story was used to demonstrate the toothbrushing procedure for all the recruited children. Parents were encouraged to show the stories to their children before or during their daily toothbrushing. Children's toothbrushing performance, oral hygiene status, and gingival health status were assessed at baseline and 6 months. Toothbrushing performance, oral hygiene, and gingival status of the recruited children were significantly improved after using social stories. Moreover, children with autism showed better oral hygiene status (P = 0.01) and better gingival status (P < 0.001) than their peers without autism. However, there were no significant differences in the toothbrushing performance among children with and without autism. The regression models indicated that the improvement of children's toothbrushing performance and oral health status were associated with children's intellectual functioning and parents' attitudes toward the usefulness of social story intervention. The principal findings suggested that social story intervention could be used to improve toothbrushing skills among children with or without autism, while it was more efficient in improving the oral hygiene status among children with autism. Autism Res 2020, 13: 666-674. © 2019 International Society for Autism Research, Wiley Periodicals, Inc. LAY SUMMARY: Toothbrushing is considered as a basic self-care practice that should be advocated in early childhood. We used social stories to train toothbrushing skills among preschoolers with special care needs. The toothbrushing skills of the recruited children were significantly improved after 6-months intervention, and children with autism showed better oral hygiene status than their peers without autism. These findings suggested that social story-based health promotion could be implemented among children with special needs.

Processing and targeting of proteins derived from polyprotein with 2A and LP4/2A as peptide linkers in a maize expression system.

In the transformation of multiple genes, gene fusion is an attractive alternative to other methods, including sexual crossing, re-transformation, and co-transformation, among others. The 2A peptide from the foot-and-mouth disease virus (FMDV) causes the co-translational "cleavage" of polyprotein and operates in a wide variety of eukaryotic cells. LP4, a linker peptide that originates from a natural polyprotein occurring in the seed of Impatiens balsamina, can be split between the first and second amino acids in post-translational processing. LP4/2A is a hybrid linker peptide that contains the first nine amino acids of LP4 and 20 amino acids of 2A. The three linkers have been used as a suitable technique to link the expression of genes in some transgenic plants, but to date the cleavage efficiency of three linkers have not been comprehensively demonstrated in the same transformation system, especially in the staple crop. To verify the functions of 2A, LP4, and LP4/2A linker peptides in transgenic maize, six fusion protein vectors that each encoded a single open reading frame (ORF) incorporating two report genes, Green Fluorescent Protein (GFP) and ß-glucuronidase (GUS), separated by 2A (or modified 2A), LP4 or LP4/2A were assembled to compare the cleavage efficiency of the three linkers in a maize transient expression system. The results demonstrated the more protein production and higher cleavage splicing efficiency with the polyprotein construct linked by the LP4/2A peptide than those of the polyprotein constructs linked by 2A or LP4 alone. Seven other fusion proteins that each encoded a single ORF incorporating two different genes GFP and Red Fluorecent Protein (RFP) with different signal peptides were assembled to study the subcellular localization of genes linked by LP4/2A. The subcellular localization experiments suggested that both types of signal peptide, co-translational and post-translational, could lead their proteins to the target localization in maize protoplast transformed by LP4/2A polyprotein construct and it implied the LP4/2A linker peptide could alleviate the inhibition of 2A processing by the carboxy-terminal region of upstream protein of 2A when translocated into the ER.