Three-dimensional analysis of hard and soft tissue changes in skeletal class II patients with high mandibular plane angle undergoing surgery.

This study aimed to study 3-dimensional (3D) changes of hard and soft tissues of skeletal class II patients after 2-jaw surgery and genioplasty. 32 adult patients diagnosed with mandibular hypoplasia who underwent 2-jaw surgery of maxillary impaction, mandibular advancement and genioplasty were enrolled. Cone-beam computed tomography and 3D stereophotogrammetry was conducted 1 week before and 6 months after surgery. Dolphin imaging software was used to establish a 3D digitizing model and 3D measurement system. Paired t-test was performed to compare the values before and after surgery. Pearson's correlation test assessed the degree of correlations between hard and soft tissue change. The mean impaction of the maxilla was 2.600 ± 3.088 mm at A. The mean advancement of the mandible was 7.806 ± 2.647 mm at B. There was a significant upward and forward movement for most landmarks of the nose and lip, while a significant decrease in nasal tip height (lateral view), upper lip height, and upper and lower vermilion height. The nose's width was significantly increased. For maxillary, Sn, Ac-r, Ac-l, and Ls demonstrated a significant correlation with A and U1 in the anteroposterior axis. However, there were no significant correlations among them in the vertical axis. For mandibular, Li demonstrated a significant correlation with L1 in the anteroposterior axis specifically for the mandible. Notably, correlations between the landmarks of the chin's hard and soft tissues were observed across all axes. The utilization of 3-D analysis facilitated a quantitative comprehension of both hard and soft tissues, thereby furnishing valuable insights for the strategic formulation of orthognathic treatment plans targeting patients with skeletal class II conditions.

Evaluation of acellular pertussis vaccine: comparisons among different strains of mice.

The current study was designed to comparatively analyse the reactions of different mouse strains in response to acellular pertussis (aP) vaccine, with attempt to further provide a reference for aP vaccine evaluation. NIH mice, ICR mice, and BALB/c mice adopted from different pharmacopoeias and studies were utilized to measure the immune protection and immunogenicity of the same batch of aP vaccine according to the Modified intracerebral challenge assay (MICA) from some Asian pharmacopoeias and the pertussis serological potency test (PTST) method from European Pharmacopoeia. Based on our results, the aP vaccine detected by NIH mice had the best potency. So the NIH mice were more suitable for detecting the immune protection of aP vaccine by the MICA method. Given that the levels of PT-IgG and FHA-IgG antibodies in ICR mice were the highest, and the levels of Th1 and Th2 cells were significantly increased (P < .01), it was more suitable for the detection of immunogenicity of aP vaccine by PSPT method. Spleen lymphocytes were stimulated by PT and FHA. And the levels of IL-4 in ICR mice and NIH mice were significantly increased, so were the levels of IL-17, IL-23, IL-27, and TNF-α in BALB/c mice. NIH mice have stronger adaptive immunity and the weakest inflammatory response, and ICR mice have enhanced adaptive immunity and inflammatory responses, both of which can be thereby used for evaluation by different pharmacopoeia methods. NIH was more suitable for the MICA method of Chinese Pharmacopoeia, and ICR for the PSPT method of European Pharmacopoeia.

Infant rhesus macaques as a non-human primate model of Bordetella pertussis infection.

BACKGROUND: The prevalent resurgence of pertussis has recently become a critical public health problem worldwide. To understand pertussis pathogenesis and the host response to both the pathogen and vaccines, a suitable pertussis animal model, particularly a non-human primate model, is necessary. Recently, a non-human primate pertussis model was successfully established with baboons. Rhesus macaques have been shown to be ideal animal models for several infectious diseases, but a model of infectious pertussis has not been established in these organisms. Studies on rhesus macaque models of pertussis were performed in the 1920s-1930s, but limited experimental details are available. Recent monkey pertussis models have not been successful because the typical clinical symptoms and transmission have not been achieved. METHODS: In the present study, infant rhesus macaques were challenged with Bordetella pertussis (B.p) using an aerosol method to evaluate the feasibility of this system as an animal model of pertussis. RESULTS: Upon aerosol infection, monkeys infected with the recently clinically isolated B.p strain 2016-CY-41 developed the typical whooping cough, leukocytosis, bacteria-positive nasopharyngeal wash (NPW), and interanimal transmission of pertussis. Both systemic and mucosal humoral responses were induced by B.p. CONCLUSION: These results demonstrate that a model of pertussis was successfully established in infant rhesus macaques. This model provides a valuable platform for research on pertussis pathogenesis and evaluation of vaccine candidates.

The effects of manufacture processes on post-translational modifications of bioactive proteins in pertussis vaccine.

Because the increasing morbidity of pertussis in all age groups worldwide, the quality of pertussis vaccines has aroused a common concern. To improve the quality of pertussis vaccine in research and production, the effects of manufacture processes on post-translational modifications (PTMs) of bioactive proteins in pertussis vaccine were investigated by a liquid chromatography quadruple - time of flight mass spectrometer (LC-Q-TOF) method in this study. The main bioactive proteins in pertussis vaccine studied include pertussis toxin (PT), pertactin (PRN) and filamentous hemagglutinin (FHA). The main manufacture processes focused are fermentation techniques, purification techniques and storage conditions. The results show that FHA and PRN are rather stable against PTM as only deamidation (Asn) was detected, which is believed to be due to their larger sizes of the bioactive proteins. For PT, however, all the manufacture processes studied have shown significant effects on types and sites of PTMs. Modifications of oxidation and demethylation (Met) occurred in the PT proteins produced by B. pertussis strain Tohama and stored in suspension in saline solution. However, they were not observed in the PT samples produced from stain CS and stored in powders. Carbamylation (Arg) on multiple sites (in S3, S4 and S5) was observed in the PT produced from 5th generation strain CS of B. pertussis. The high abundance ratio of carbamylation modification was potentially a negative effect on the detoxification of PT, since unmodified Lys was the active site for detoxification. The results obtained in this study provide information for making protection strategies against PTMs in pertussis vaccine in manufacture and storage.

A preliminary study on the application of PspA as a carrier for group A meningococcal polysaccharide.

This study aimed to explore the feasibility of pneumococcal surface protein A (PspA) as a carrier protein. Three recombinant pneumococcal surface proteins from three different clades were expressed by the prokaryotic expression system and conjugated to group A meningococcal polysaccharide (GAMP) to generate three polysaccharide-protein conjugates. The conjugates, unconjugated proteins, GAMP, and GAMP-TT vaccine bulk (used as positive control) were immunized into mice, and their immune effects were assessed by the methods of enzyme-linked immunosorbent assay (ELISA), flow cytometry (FCM), and serum bactericidal assay (SBA). The results showed that the polysaccharide-protein conjugates could produce higher levels of anti-GAMP IgG titers (P < 0.05), higher ratios of Th1/Th2 (P < 0.05), and higher levels of serum bactericidal activity (P < 0.05), compared with the unconjugated GAMP. The conjugation of PspAs to GAMP also enhanced the anti-PspA responses compared with unconjugated PspAs except for PspA3. In conclusion, the results indicated that the three PspAs were appropriate carrier proteins, as demonstrated by the characteristics of T-cell dependent responses to the GAMP, and might protect against group A of epidemic cerebrospinal meningitis.

Genetic diversity and population dynamics of Bordetella pertussis in China between 1950-2007.

Pertussis is an acute respiratory infectious disease caused by the bacterium Bordetella pertussis. Although pertussis vaccination was introduced in the 1960s, pertussis is still an endemic disease in China. To better understand the genetic diversity of the Chinese B. pertussis population, we characterized 115 clinical isolates obtained in China during 1950-2007 using multilocus variable-number tandem repeat analysis (MLVA). Forty-six different B. pertussis MLVA profiles (MTs) were identified, of which 13 were new MTs. Analysis using a minimum-spanning tree showed that distinct MTs were prevalent during different periods, suggesting that a dynamic change in B. pertussis MTs occurred over time in China. The predominant MTs in recent isolates from China were different from those of many developed countries. A decreasing trend in genetic diversity of the B. pertussis population was observed following the introduction of pertussis vaccines. Similar to the pertactin 2 (prn2) allele, the novel pertussis toxin promoter (ptxP3) allele first emerged in 2000, but unlike trends elsewhere, ptxP1 remained predominant among the isolates, further reflecting the unique temporal trends in the B. pertussis population in China. Our results suggest that temporal changes in the B. pertussis population may be closely associated with vaccination coverage and the vaccine types used. These data may lead to an improved understanding of the virulence mechanism of B. pertussis and facilitate new strategies for controlling this infectious disease.

Whole-genome sequencing reveals the effect of vaccination on the evolution of Bordetella pertussis.

Herd immunity can potentially induce a change of circulating viruses. However, it remains largely unknown that how bacterial pathogens adapt to vaccination. In this study, Bordetella pertussis, the causative agent of whooping cough, was selected as an example to explore possible effect of vaccination on the bacterial pathogen. We sequenced and analysed the complete genomes of 40 B. pertussis strains from Finland and China, as well as 11 previously sequenced strains from the Netherlands, where different vaccination strategies have been used over the past 50 years. The results showed that the molecular clock moved at different rates in these countries and in distinct periods, which suggested that evolution of the B. pertussis population was closely associated with the country vaccination coverage. Comparative whole-genome analyses indicated that evolution in this human-restricted pathogen was mainly characterised by ongoing genetic shift and gene loss. Furthermore, 116 SNPs were specifically detected in currently circulating ptxP3-containing strains. The finding might explain the successful emergence of this lineage and its spread worldwide. Collectively, our results suggest that the immune pressure of vaccination is one major driving force for the evolution of B. pertussis, which facilitates further exploration of the pathogenicity of B. pertussis.

Seroprevalence of pertussis in China: need to improve vaccination strategies.

Pertussis remains an important cause of infant death worldwide and is an ongoing public health concern even in countries with high vaccination coverage. A cross-sectional seroepidemiological study was undertaken to estimate true incidence rates and gain further insight into the epidemiology and burden of pertussis in China. During 2011, a total of 1080 blood samples were obtained from healthy individuals between 0 and 86 y of age in Zhengzhou, Central China. Serum IgG antibodies against pertussis toxin (PT) and filamentous hemagglutinin (FHA) were measured quantitatively using ELISA. The results showed that the geometric mean titers of PT and FHA IgG were 6.48 IU/mL (95% CI: 5.70-7.41 IU/mL) and 11.39 IU/mL (95% CI: 10.22-12.87 IU/mL) among subjects less than 4 y of age, indicating that pertussis antibody levels were low despite high vaccination coverage. Of the 850 subjects≥4 y of age, 56 (6.6%) had anti-PT IgG titers above 30 IU/mL, and 11 (1.3%) had antibodies titers above 80 IU/mL. The estimated age-specific incidence of infection with B. pertussis revealed a peak incidence in the 31 to 40 y age group, followed by the 41 to 60 y age group. Taken together, these results indicate that pertussis is common in Chinese subjects in Zhengzhou, especially in adults, suggesting that the disease burden is underestimated in China. Therefore, our study stresses the importance of strengthening the diagnostic capacity and improving surveillance system for delineating current epidemiological profiles of pertussis. Most importantly, it may be advisable to re-evaluate the current Chinese pertussis immunization schedule and implement to booster doses for older children, adolescents and adults.

Acellular pertussis vaccines in China.

In China, whole-cell pertussis (Pw) vaccines were produced in the early 1960s and acellular pertussis (Pa) vaccines were introduced in 1995. Pa vaccines have now almost completely replaced Pw vaccines in the national immunization program. To strengthen the regulation of vaccines used in China, a vaccine lot release system was established in 2001 and Pa vaccines have been included in the system since 2006. This paper mainly described the current status of production and the quality control measures in place for Pa vaccines; and analyses quality control test data accumulated between 2006 and 2010.

Safety and immunogenicity of a diphtheria, tetanus, acellular pertussis and Haemophilus influenzae Type b combination vaccine compared with separate administration of licensed equivalent vaccines in Chinese infants and toddlers for primary and booster immunization.

To evaluate the safety and immunogenicity of a diphtheria, tetanus, acellular pertussis and Haemophilus infuenzae Type b (DTaP/Hib) combination vaccine first developed by a Chinese manufacturer, a randomized, two-stage, parallel controlled, single center clinical trial was conducted in Dafeng, Jiangsu Province of China. A total of 720 infants were enrolled and randomly assigned to two groups with a 2:1 allocation. In Stage I, 480 subjects in Group T were administered with 3 doses of the DTaP/Hib vaccine at 3, 4 and 5 months of age, respectively, while 240 subjects in Group C received separate licensed DTaP vaccine and Hib conjugate vaccine on the same schedule. In Stage II, 633 primed toddlers (431 of Group T and 202 of Group C) were given a booster dose at 18 months of age. Sera samples were collected at pre-dose 1, 4 weeks post-dose 3, pre-dose 4 and 4 weeks post-dose 4, respectively. Levels of protective antibodies were measured by Enzyme Linked Immunoadsorbent Assay (ELISA). Immunogenicity was evaluated with regard to geometric mean concentrations (GMCs), seroconversion rates and seroprotection rates of the antibodies. Solicited adverse reactions were recorded for 3 days after each dose; unsolicited adverse events and serious adverse events were monitored for 28 days after vaccination. Results showed that seroconversion rates of anti-pertussis toxoid (PT), anti-filamentous hemagglutinin (FHA), anti-diphtheria toxoid (DT), anti-tetanus toxid (TT) and anti-polyribosyl-ribitol-phosphate (PRP) in Group T (Stage I: 98.06%, 97.33%, 100%, 100%, 98.79%; Stage II: 99.18%, 83.42%, 99.18%, 63.32%, 85.05%) were comparable to that of Group C (Stage I: 95.26%, 93.16%, 100%, 100%, 98.42%; Stage II: 98.89%, 83.89%, 98.33%, 53.89%, 76.67%). Nearly 100% of the subjects in both groups achieved seroprotective levels of anti-DT (> or = 0.1IU/ml), anti-TT (> or = 0.1IU/ml) and anti-PRP (> or = 0.15 microg/ml) after primary and booster vaccination. The frequencies of local induration, swelling and redness as well as general reactions such as fever, diarrhea and anaphylaxis were low and acceptable in both groups. In conclusion, the DTaP/Hib vaccine was demonstrated to be non-inferior to the control vaccines on safety and immunogenicity. There could be a bright future for the DTaP/Hib vaccine to be widely used in the universal vaccination of Chinese children.

Production and characterization of recombinant pertactin, fimbriae 2 and fimbriae 3 from Bordetella pertussis.

BACKGROUND: Bordetella pertussis is a causative agent of pertussis or whooping cough in humans. Pertactin (Prn), fimbriae 2 (Fim2) and fimbriae 3 (Fim3) of B. pertussis are important virulence factors and immunogens which have been included in some acellular pertussis vaccines. In this present study, we cloned, expressed and purified Prn, Fim2 and Fim3, respectively. The immunogenicity and protective efficacy of the three recombinant proteins (rPrn, rFim2 and rFim3) were investigated in mouse model. RESULTS: Three recombinant proteins with amount of 12 to 25 mg/L were produced. Compared to the control mice only immunized with adjuvant, serum IgG antibody responses were significantly induced in the mice immunized with rPrn, rFim2 or rFim3 (P < 0.001 for all three proteins). Furthermore, T cell responses characteristic of increased production of IL-2 and TNF-alpha (only for rPrn) were elicited in the mice immunized with the three proteins (P < 0.05 for all three proteins). Immunization with rPrn, but not with rFim2 or rFim3, significantly enhanced clearance of bacteria in the lungs of mice after intranasal challenge with B. pertussis (P < 0.05). When tested in a lethal intracerebral infection model, certain protection was observed in mice immunized with rPrn. CONCLUSIONS: We have developed an efficient method to produce large amounts of rPrn, rFim2, and rFim3 from B. pertussis. The three recombinant proteins induced both humoral and cellular immune responses in mice. Immunization with rPrn also conferred protection against pertussis in mouse infection models. Our results indicated that the recombinant proteins still retain their immunological properties and highlighted the potential of the recombinant proteins for the future development of the B. pertussis vaccines.

Construction and characterization of a novel superantigen fusion protein: bFGF/SEB.

BACKGROUND: As one of the bacterial superantigens, Staphylococcal enterotoxins (SEs) are potent activators of T cells, especially for those expressing T cell receptor V(beta) chains, and can induce the production of cytokines such as IFN-gamma, TNF-alpha, IL-1, IL-2, IL-6, IL-12, etc. Thus, SEs could be used in tumor-targeting therapy when cooperated with the vectors that can specifically recognize the tumor cells. MATERIALS AND METHODS: The coding sequences of Staphylococcal enterotxin B (SEB) was amplified and fused with human basic fibroblast growth factor (bFGF). Recombinant protein SEB and fusion protein bFGF/SEB were expressed and purified. The biological activity was detected, including splenocytes proliferation, cytokine production, and cytotoxicity in tumor cells in vitro. In addition, the binding of bFGF/SEB with tumor cells and the tumor cell apoptosis were also tested by immunofluorescent technique. RESULTS: The fusion protein bFGF/SEB had similar biological activities compared with natural SEA and recombinant SEB, including tumor-inhibition ratio. CONCLUSION: The recombinant bFGF/SEB-fusion protein was shown to retain the superantigenic activity of SEB, and might be a novel promising immunotherapeutic agent for the treatment of some carcinomas.