Cross-sectional study of C1–S5 vertebral bodies in human fetuses

Introduction

Knowledge on the normative spinal growth is relevant in the prenatal detection of its abnormalities. The present study determines the height, transverse and sagittal diameters, cross sectional area, and volume of individual C1–S5 vertebral bodies.

Material and methods

Using the methods of computed tomography (CT), digital image analysis, and statistics, the size of C1–S5 vertebral bodies in 55 spontaneously aborted human fetuses aged 17–30 weeks was examined.

Results

All the 5 examined parameters changed significantly with gestational age (p < 0.01). The mean height of vertebral bodies revealed an increase from the atlas (2.39 ±0.54 mm) to L2 (4.62 ±0.97 mm), stabilized through L3–L4 (4.58 ±0.92 mm, 4.61 ±0.84 mm), and then was decreasing to S5 (0.43 ±1.06 mm). The mean transverse diameter of vertebral bodies was increasing from the atlas (1.20 ±1.96 mm) to L1 (6.24 ±1.46 mm), so as to stabilize through L2–L3 (6.12 ±1.65, 6.12 ±1.61 mm), and finally was decreasing to S5 (0.26 ±0.96 mm). There was an increase in sagittal diameter of vertebral bodies from the atlas (0.82 ±1.34 mm) to T7 (4.76 ±0.85 mm), its stabilization for T8–L4 (4.73 ±0.86 mm, 4.71 ±1.02 mm), and then a decrease in values to S5 (0.21 ±0.75 mm) was observed. The values for cross-sectional area of vertebral bodies were increasing from the atlas (2.95 ±5.25 mm²) to L3 (24.92 ±11.07 mm²), and then started decreasing to S5 (0.48 ±2.09 mm²). The volumetric growth of vertebral bodies was increasing from the atlas (8.60 ±16.40 mm³) to L3 (122.16 ±74.73 mm³), and then was decreasing to S5 (1.60 ±7.00 mm³).

Conclusions

There is a sharp increase in size of fetal vertebral bodies between the atlas and the axis, and a sharp decrease in size within the sacral spine. In human fetuses the vertebral body growth is characterized by maximum values in sagittal diameter for T7, in transverse diameter for L1, in height for L2, and in both cross-sectional area and volume for L3.     

Cross-sectional study of the neural ossification centers of vertebrae C1–S5 in the human fetus

Purpose

An understanding of the normal evolution of the spine is of great relevance in the prenatal detection of spinal abnormalities. This study was carried out to estimate the length, width, cross-sectional area and volume of the neural ossification centers of vertebrae C1–S5 in the human fetus.

Materials and methods

Using the methods of CT (Biograph mCT), digital-image analysis (Osirix 3.9) and statistics (the one-way ANOVA test for paired data, the Kolmogorov–Smirnov test, Levene’s test, Student’s t test, the one-way ANOVA test for unpaired data with post hoc RIR Tukey comparisons) the size for the neural ossification centers throughout the spine in 55 spontaneously aborted human fetuses (27 males, 28 females) at ages of 17–30 weeks was studied.

Results

The neural ossification centers were visualized in the whole pre-sacral spine, in 74.5 % for S1, in 61.8 % for S2, in 52.7 % for S3, and in 12.7 % for S4. Neither male–female nor right–left significant differences in the size of neural ossification centers were found. The neural ossification centers were the longest within the cervical spine. The maximum values referred to the axis on the right, and to C5 vertebra on the left. There was a gradual decrease in length for the neural ossification centers of T1–S4 vertebrae. The neural ossification centers were the widest within the proximal thoracic spine and narrowed bi-directionally. The growth dynamics for CSA of neural ossification centers were found to parallel that of volume. The largest CSAs and volumes of neural ossification centers were found in the C3 vertebra, and decreased in the distal direction.

Conclusions

The neural ossification centers show neither male–female nor right–left differences. The neural ossification centers are characterized by the maximum length for C2–C6 vertebrae, the maximum width for the proximal thoracic spine, and both the maximum cross-sectional area and volume for C3 vertebra. There is a sharp decrease in size of the neural ossification centers along the sacral spine. A decreasing sequence of values for neural ossification centers along the spine from cervical to sacral appears to parallel the same direction of the timing of ossification. The quantitative growth of the neural ossification centers is of potential relevance in the prenatal diagnosis and monitoring of achondrogenesis, caudal regression syndrome, diastematomyelia and spina bifida.     

Diffuse distribution of bone cement in percutaneous vertebroplasty reduces the incidence of refracture of adjacent vertebral bodies北大核心

BACKGROUND: Percutaneous vertebroplasty is one of the effective treatments for osteoporotic vertebral compression fractures. The distribution of bone cement in the vertebral body will influence the clinical efficacy. OBJECTIVE: To investigate the influence of the distribution of bone cement in percutaneous vertebroplasty on the early postoperative outcomes after osteoporotic vertebral compression fractures. METHODS: A total of 132 patients, including 52 males and 80 females, with a mean age of (76.3±8.2) years who diagnosed as osteoporotic vertebral compression fractures in First Hospital of Shanxi Medical University from January 2017 to June 2019 were enrolled. All patients were divided into underdispersion group (n=54) and diffuse distribution group (n=78) according to the pattern of cement distribution on postoperative X-rays. Visual analogue scale scores postoperatively, the height of anterior edge and midline of injured vertebrae, Cobb angle of local kyphosis and the incidence of complications after 1-year follow-up were compared between the two groups. The trial was performed in accordance with the relevant ethical requirements for research in the First Hospital of Shanxi Medical University. RESULTS AND CONCLUSION: (1) The visual analogue scale scores were lower at 2 days, 1 and 6 months after operation than those before operation in the two groups (P < 0.05). The visual analogue scale scores at 1 month after operation in the underdispersion group were higher than those in the diffuse distribution group (P < 0.05), and there was no significant difference between the two groups at 2 days and 6 months after operation (P > 0.05). (2) Cobb angle and vertebral body height improved in both groups postoperatively compared to preoperatively (P < 0.05). The difference in Cobb angle and vertebral body height was not statistically significant between the two groups (P > 0.05). (3) The rate of bone cement infiltration was not significantly different between the two groups (P > 0.05). Incidence of recurrent fractures in adjacent vertebrae was lower in the diffuse distribution group than that in the underdispersion group (P < 0.05). (4) These findings confirm that percutaneous vertebroplasty is an effective treatment for osteoporotic vertebral compression fractures. The distribution of bone cement in the vertebral body will affect the surgical results. The bone cement is diffusely distributed, which may reduce the incidence of recurrent fractures in adjacent vertebrae. © 2022, Publishing House of Chinese Journal of Tissue Engineering Research. All rights reserved.     

Characterization of the annulus fibrosus–vertebral body interface: identification of new structural features

Current surgical treatments for degenerative intervertebral disc disease do not restore full normal spinal movement. Tissue engineering a functional disc replacement may be one way to circumvent this limitation, but will require an integration of the different tissues making up the disc for this approach to be successful. Hence, an in-depth characterization of the native tissue interfaces, including annulus insertion into bone is necessary, as knowledge of this interface is limited. The objective of this study was to characterize the annulus fibrosus–vertebral bone (AF–VB) interface in immature (6–9 months old) and mature (18–24 months old) bovine discs, as well as to define these structures for normal adult human (22 and 45 years old) discs. Histological assessment showed that collagen fibers in the inner annulus, which are predominantly type II collagen, all appear to insert into the mineralized endplate zone. In contrast, some of the collagen fibers of the outer annulus, predominantly type I collagen, insert into this endplate, while other fibers curve laterally, at an ∼ 90 ° angle, to the outer aspect of the bone, and merge with the periosteum. This is seen in both human and bovine discs. Where the AF inserts into the calcified zone of the AF–VB interface, it passes through a chondroid region, rich in type II collagen and proteoglycans. Annulus cells (elongated cells that are not surrounded by proteoglycans) are present at this interface. This cartilage zone is evident in both human and bovine discs. Type X collagen and alkaline phosphatase are localized to the interface region. Age-associated differences in bovine spines are observed when examining the interface thickness and the matrix composition of the cartilaginous endplate, as well as the thickness of the mineralized endplate. These findings will assist with the design of the AF–VB interface in the tissue engineered disc.     

The sphingosine 1-phosphate receptor S1P₂ maintains the homeostasis of germinal center B cells and promotes niche confinement

Mice deficient in sphingosine 1-phosphate receptor type 2 (S1P(2)) develop diffuse large B cell lymphoma. However, the role of S1P(2) in normal germinal center (GC) physiology is unknown. Here we show that S1P(2)-deficient GC B cells outgrew their wild-type counterparts in chronically established GCs. We found that antagonism of the kinase Akt mediated by S1P(2) and its downstream mediators Gα(12), Gα(13) and p115RhoGEF regulated cell viability and was required for growth control in chronically proliferating GCs. Moreover, S1P(2) inhibited GC B cell responses to follicular chemoattractants and helped confine cells to the GC. In addition, S1P(2) overexpression promoted the centering of activated B cells in the follicle. We suggest that by inhibiting Akt activation and migration, S1P(2) helps restrict GC B cell survival and localization to an S1P-low niche at the follicle center.     

Decomposition and entomological colonization of charred bodies – a pilot study

Aim

To use forensic entomological approach to estimate the post mortem interval (PMI) in burnt remains.

Methods

Two experiments were performed in a field in the outskirts of Milan, in winter and summer 2007. Four 60-kg pigs were used: two for each experiment. One pig carcass was burnt until it reached the level 2-3 of the Glassman-Crow scale and the not-burnt carcass was used as a control. In order to describe the decomposition process and to collect the data useful for minimum PMI estimation, macroscopic, histological, and entomological analyses were performed.

Results

In the winter part of the experiment, the first insect activity on the burnt carcass began in the third week (Calliphora vomitoria) and at the beginning of the fourth week an increase in the number of species was observed. In the summer part, adult flies and first instar maggots (Phormia regina) appeared a few minutes/hours after the carcass exposure. Both in winter and summer, flies belonging to the first colonization wave (Calliphoridae) appeared on burnt and control pigs at the same time, whereas other species (Diptera and Coleoptera) appeared earlier on burnt pigs.

Conclusion

In forensic practice, burnt bodies are among the most neglected fields of entomological research, since they are supposed to be an inadequate substratum for insect colonization. Entomological approach for PMI estimation proved to be useful, although further studies on larger samples are needed.Estimation of the post mortem interval (PMI) plays an important role in forensic investigation. In the early post mortem period, PMI can be estimated by temperature-based methods, but when decomposition begins, this estimation can be influenced by several variables (1,2). In addition, in cases of concealment, body dismemberment, explosion, and burning there is no standardized method based on experimental studies for deriving time since death from morphological characteristics of the corpse.Entomological approach is a well known and widely accepted method to estimate the minimum PMI (3). However, in the literature there are only a few cases referring to charred bodies (4-8). Gruenthal et al (9) found dung fly Scathophaga stercoraria, larvae of Calliphora vicina and Calliphora vomitoria, and immature beetle forms, not further identified in 24 pig carcasses charred up to Glassman Crow scale-1 (GCS-1) for the head, neck, limbs, and CGS 2 for the torso. Catts and Goff (6) observed a few days’ delay in the arrival of blowflies on a corpse burnt and charred inside an open-topped metal drum, and a week’s delay in the case of a pig burnt inside a car that was set afire (6). Introna et al (4) also highlighted that burnt flesh delayed the arrival of blowflies. Due to the relevant lack of literature, the aim of our study was to report the results of an experimental approach to burnt bodies using pigs (Sus scrofa) as models.     

Osteophytes on the zygapophyseal (facet) joints of the cervical spine (C3–C7): A skeletal study

Previous studies have reported that osteophytes in the cervical vertebrae may cause immobility, neck stiffness, osteoarthritis, headaches, nerve entrapment syndromes, and compression of the vertebral artery. Our objective was to explore the osteophytes' expression on zygapophyseal joints C3–C7. This is a cross-sectional observational skeletal study. The study sample comprised 273 human skeletons of both sexes, aged 20–93, housed at the Natural History Museum, OH, USA. A grading system assessed the presence and severity of osteophytosis on the zygapophyseal joints. The chi-square test (SPSS 25.0) examined the association between osteophytes and demographic factors. The level of significance (α) was set at .05. The highest prevalence of osteophytes was found on C5 vertebra, the lowest on C7. On vertebrae C3, C4, C6, the rate of moderate and severe osteophytes found on the superior and inferior facets were comparable. Moderate and severe degrees of osteophytes were observed more frequently on the superior facets, whereas, on vertebra C7, osteophytes were found on the inferior facet joints. Osteophytes' prevalence was significantly higher in the elderly compared to the younger population. Osteophytes in the C3–C7 zygapophyseal joints are age-dependent. No significant sex and ethnic differences were observed. Vertebra C5 was most prone to develop osteophytes, most probably due to its location in the cervical lordotic peak, C5 in the superior ZF; C7 in the inferior ZF are significant (p = .05). The zygapophyseal joints of C7 were least frequent overall, yet, the C7 inferior facets had significantly more moderate–severe osteophytes compared to other cervical vertebrae.     

The trabecular architecture of the superior articular process of the lumbar spine (L2–S1)

The role of the facet joint in low back pain has gained public attention lately. The objective of our study was to investigate whether there is any difference in the adaptation of the cancellous bone in the superior articular process depending on the specific stress condition in different levels of the spine. Therefore, the trabecular structure of the superior articular processes of L2 and S1 of 15 cadavers (aged 63–100 years) were studied using μCT (micro-computer tomography). Each sample was divided into five sections, each of which containing 20% of the slices. The following structure parameters were compared between L2 and S1 as well as within each process; bone-volume-fraction (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), structure-model-index (SMI) and degree of anisotropy (DA). Statistically significant differences were observed between L2 and S1 for the BV/TV, SMI, Tb.Th and Tb.N in superior 2 sections. BV/TV, Tb.Th and Tb.N were higher in S1 than in L2. The SMI is lower, and even negative in S1 compared to L2, showing a more plate-like structure. Within the articular process all structure parameters show a similar distribution in L2 and S1. BV/TV, Tb.N and DA decreased from cranial to caudal while Tb.Th was highest in the most cranial and caudal sections, with the lowest value in the middle. The SMI, on the other hand, increased from cranial to caudal displaying more rod-like structures. These results can be explained by the different stress the processes of the different spinal levels are exposed to as well as the specific motion patterns of the facet joint. The processes of the os sacrum are exposed to a higher axial and ventral load due to their location and the lumbosacral flexion. In addition the upper sections of each process experience higher stress peaks than the lower ones. Therefore, this study shows the material distribution within the cancellous bone adapts to these specific stress conditions the facet joints are exposed to.     

Sequence analysis of the 5′ third of glycoprotein C gene of South American bovine herpesviruses 1 and 5

Bovine herpesviruses 1 (BoHV-1) and 5 (BoHV-5) share high genetic and antigenicsimilarities, but exhibit marked differences in tissue tropism and neurovirulence.The amino-terminal region of glycoprotein C (gC), which is markedly different in eachof the viruses, is involved in virus binding to cellular receptors and ininteractions with the immune system. This study investigated the genetic andantigenic differences of the 5′ region of the gC (5′ gC) gene (amino-terminal) ofSouth American BoHV-1 (n=19) and BoHV-5 (n=25) isolates. Sequence alignments of 374nucleotides (104 amino acids) revealed mean similarity levels of 97.3 and 94.2% amongBoHV-1 gC (gC1), respectively, 96.8 and 95.6% among BoHV-5 gC (gC5), and 62 and 53.3%between gC1 and gC5. Differences included the absence of 40 amino acid residues (27encompassing predicted linear epitopes) scattered throughout 5′ gC1 compared to 5′gC5. Virus neutralizing assays testing BoHV-1 and BoHV-5 antisera against eachisolate revealed a high degree of cross-neutralization between the viruses, yet someisolates were neutralized at very low titers by heterologous sera, and a few BoHV-5isolates reacted weakly with either sera. The virus neutralization differencesobserved within the same viral species, and more pronounced between BoHV-1 andBoHV-5, likely reflect sequence differences in neutralizing epitopes. These resultsdemonstrate that the 5′ gC region is well conserved within each viral species but isdivergent between BoHV-1 and BoHV-5, likely contributing to their biological andantigenic differences.     

Predictors of the quality of cardiovascular prevention �C a multilevel cross-sectional study

Aim

To attempt to develop a model of predictors for quality of the process of cardiovascular prevention in patients at high risk of cardiovascular disease (CVD).

Methods

We formed a random sample of patients from a stratified sample of 36 family practice registers of patients at high risk of CVD without diabetes and without established CVD. Data were gathered by chart audit and questionnaires about patient and practice characteristics. We defined the process of care as a dependent variable by principle component analysis and tested the relationship of the process with several independent variables (family physicians’, patients’, and practice characteristics). To study the effects of independent variables (predictors) on the process of care we carried out multilevel regression analysis with the patients constituting the lower level and nested within the family physician/practice (the second level).

Results

Multilevel regression analysis included 645 patients from 36 practices (74.1% from the final sample). Patients’ characteristics that predicted the higher-quality process of CVD prevention were younger age (t = -4.94, 95% confidence interval [CI] -0.018 to -0.008) and lower socioeconomic status (t = -2.18, 95%CI -0.195 to -0.010). Practice characteristics that predicted the higher-quality process of CVD prevention were smaller practice size (t = 2.83, 95% CI 0.063 to 1.166), a good information system for CVD prevention (t = 3.15, 95% CI 0.030 to 0.282), and the organization of education on CVD prevention (t = 3.19, 95%CI 0.043 to 0.380).

Conclusion

This study shows that the quality of cardiovascular prevention could be measured as a composite outcome and future studies should further develop this approach and test the impact of several practice/patient characteristics on the quality of CVD prevention with the international data.Prevention of cardiovascular diseases (CVD) is an important task for family physicians. While patients with a low risk of CVD profit mostly from public health activities, high risk patients also need preventive activities provided by their family physicians (1,2). In the countries with a national program of CVD prevention (Slovenia is one of them), these activities and procedures can be highly standardized (3) and, therefore, should be easily measurable. The Slovenian national preventive program for CVD was launched in 2001 and requires preventive check-ups for the defined age groups of patients (women from 45 to 70 years and men from 35 to 65 years). Eighty percent of the target group in every practice needs to go through the program in 5 years, and a register of high risk patients is created in each practice and collected on the national level. The preventive activities consist of two parts: the first part includes a health check-up with determination of risk factors (information on life-style, clinical exam, laboratory tests of lipids and fasting blood glucose) and the second part includes the referral of patients at high risk to preventive workshops, for example for healthy weight reduction, smoking cessation, etc.Although there is some evidence on several isolated aspects of CVD prevention in Slovenia (4-6), a comprehensive and systematic approach for measuring its quality and actual outcomes is still not available. Therefore, we aimed to develop an integral statistically evaluated presentation of the process of cardiovascular prevention and determine the variables that influence it. Post-hoc analyses were performed on patients at a high risk for coronary diseases using Slovenian data from the international EPA-Cardio study, a cross-sectional study conducted in 9 European countries that had developed quality indicators for cardiovascular prevention on the international level (7) and evaluated the quality of cardiovascular prevention for high-risk patients (8).     

Functional study of the different haplotypes cDNA in the coding region of CⅡTA gene

Objective To study the function of 4 different haplotypes cDNA which are constructed by two non-homonymy single nueleotide polymorphism (SNP) sites C19170G (Leu45Val) and C30799G (Ala500Gly) in the coding region of human CⅡTA gene. Methods HeLa cells were transfeeted with eu-karyotic expression vectors containing four different haplotypes cDNA. C Ⅱ TA mRNA and HLA classⅡanti-gen (HLA-DR, DP, DQ) were respectively detected by RT-PCR and indirect cell immunofluoreseence tech-nique in the untransfected and transfeeted with four eukaryotic expression vectors and empty vectors HeLa cells. The quantity of HLA classⅡ antigen were analyzed by flow eytometry. Results No expression of CⅡTA mRNA and HLA class Ⅱ antigen were observed on original HeLa cells and empty vector transfected cells. CⅡTA mRNA expression was emerged, and the expression of HLA class Ⅱ antigen were observed in the HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA. And there were not significantly different with the levels of HLA class Ⅱ antigen expression among HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA ( P > 0.05 ). Con-dusion The SNP of Chinese at the sites C19170G(Leu45Val) and C30799G(Ala500Gly) in the coding site of C Ⅱ TA gene did not influence capability of CⅡTA trans-aetivating HLA class Ⅱgene expression.     

Functional study of the different haplotypes cDNA in the coding region of CⅡTA gene

Objective To study the function of 4 different haplotypes cDNA which are constructed by two non-homonymy single nueleotide polymorphism (SNP) sites C19170G (Leu45Val) and C30799G (Ala500Gly) in the coding region of human CⅡTA gene. Methods HeLa cells were transfeeted with eu-karyotic expression vectors containing four different haplotypes cDNA. C Ⅱ TA mRNA and HLA classⅡanti-gen (HLA-DR, DP, DQ) were respectively detected by RT-PCR and indirect cell immunofluoreseence tech-nique in the untransfected and transfeeted with four eukaryotic expression vectors and empty vectors HeLa cells. The quantity of HLA classⅡ antigen were analyzed by flow eytometry. Results No expression of CⅡTA mRNA and HLA class Ⅱ antigen were observed on original HeLa cells and empty vector transfected cells. CⅡTA mRNA expression was emerged, and the expression of HLA class Ⅱ antigen were observed in the HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA. And there were not significantly different with the levels of HLA class Ⅱ antigen expression among HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA ( P > 0.05 ). Con-dusion The SNP of Chinese at the sites C19170G(Leu45Val) and C30799G(Ala500Gly) in the coding site of C Ⅱ TA gene did not influence capability of CⅡTA trans-aetivating HLA class Ⅱgene expression.     

BLOC1S5 pathogenic variants cause a new type of Hermansky–Pudlak syndrome

PurposeHermansky–Pudlak syndrome (HPS) is characterized by oculocutaneous albinism, excessive bleeding, and often additional symptoms. Variants in ten different genes have been involved in HPS. However, some patients lack variants in these genes. We aimed to identify new genes involved in nonsyndromic or syndromic forms of albinism.MethodsTwo hundred thirty albinism patients lacking a molecular diagnosis of albinism were screened for pathogenic variants in candidate genes with known links to pigmentation or HPS pathophysiology.ResultsWe identified two unrelated patients with distinct homozygous variants of the BLOC1S5 gene. Patients had mild oculocutaneous albinism, moderate bleeding diathesis, platelet aggregation deficit, and a dramatically decreased number of platelet dense granules, all signs compatible with HPS. Functional tests performed on platelets of one patient displayed an absence of the obligate multisubunit complex BLOC-1, showing that the variant disrupts BLOC1S5 function and impairs BLOC-1 assembly. Expression of the patient-derived BLOC1S5 deletion in nonpigmented murine Bloc1s5^-/- melan-mu melanocytes failed to rescue pigmentation, the assembly of a functional BLOC-1 complex, and melanosome cargo trafficking, unlike the wild-type allele.ConclusionMutation of BLOC1S5 is disease-causing, and we propose that BLOC1S5 is the gene for a new form of Hermansky–Pudlak syndrome, HPS-11.     

Functional study of the different haplotypes cDNA in the coding region of CⅡTA gene

Objective To study the function of 4 different haplotypes cDNA which are constructed by two non-homonymy single nueleotide polymorphism (SNP) sites C19170G (Leu45Val) and C30799G (Ala500Gly) in the coding region of human CⅡTA gene. Methods HeLa cells were transfeeted with eu-karyotic expression vectors containing four different haplotypes cDNA. C Ⅱ TA mRNA and HLA classⅡanti-gen (HLA-DR, DP, DQ) were respectively detected by RT-PCR and indirect cell immunofluoreseence tech-nique in the untransfected and transfeeted with four eukaryotic expression vectors and empty vectors HeLa cells. The quantity of HLA classⅡ antigen were analyzed by flow eytometry. Results No expression of CⅡTA mRNA and HLA class Ⅱ antigen were observed on original HeLa cells and empty vector transfected cells. CⅡTA mRNA expression was emerged, and the expression of HLA class Ⅱ antigen were observed in the HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA. And there were not significantly different with the levels of HLA class Ⅱ antigen expression among HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA ( P > 0.05 ). Con-dusion The SNP of Chinese at the sites C19170G(Leu45Val) and C30799G(Ala500Gly) in the coding site of C Ⅱ TA gene did not influence capability of CⅡTA trans-aetivating HLA class Ⅱgene expression.     

Functional study of the different haplotypes cDNA in the coding region of CⅡTA gene

Objective To study the function of 4 different haplotypes cDNA which are constructed by two non-homonymy single nueleotide polymorphism (SNP) sites C19170G (Leu45Val) and C30799G (Ala500Gly) in the coding region of human CⅡTA gene. Methods HeLa cells were transfeeted with eu-karyotic expression vectors containing four different haplotypes cDNA. C Ⅱ TA mRNA and HLA classⅡanti-gen (HLA-DR, DP, DQ) were respectively detected by RT-PCR and indirect cell immunofluoreseence tech-nique in the untransfected and transfeeted with four eukaryotic expression vectors and empty vectors HeLa cells. The quantity of HLA classⅡ antigen were analyzed by flow eytometry. Results No expression of CⅡTA mRNA and HLA class Ⅱ antigen were observed on original HeLa cells and empty vector transfected cells. CⅡTA mRNA expression was emerged, and the expression of HLA class Ⅱ antigen were observed in the HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA. And there were not significantly different with the levels of HLA class Ⅱ antigen expression among HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA ( P > 0.05 ). Con-dusion The SNP of Chinese at the sites C19170G(Leu45Val) and C30799G(Ala500Gly) in the coding site of C Ⅱ TA gene did not influence capability of CⅡTA trans-aetivating HLA class Ⅱgene expression.     

Functional study of the different haplotypes cDNA in the coding region of CⅡTA gene

Objective To study the function of 4 different haplotypes cDNA which are constructed by two non-homonymy single nueleotide polymorphism (SNP) sites C19170G (Leu45Val) and C30799G (Ala500Gly) in the coding region of human CⅡTA gene. Methods HeLa cells were transfeeted with eu-karyotic expression vectors containing four different haplotypes cDNA. C Ⅱ TA mRNA and HLA classⅡanti-gen (HLA-DR, DP, DQ) were respectively detected by RT-PCR and indirect cell immunofluoreseence tech-nique in the untransfected and transfeeted with four eukaryotic expression vectors and empty vectors HeLa cells. The quantity of HLA classⅡ antigen were analyzed by flow eytometry. Results No expression of CⅡTA mRNA and HLA class Ⅱ antigen were observed on original HeLa cells and empty vector transfected cells. CⅡTA mRNA expression was emerged, and the expression of HLA class Ⅱ antigen were observed in the HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA. And there were not significantly different with the levels of HLA class Ⅱ antigen expression among HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA ( P > 0.05 ). Con-dusion The SNP of Chinese at the sites C19170G(Leu45Val) and C30799G(Ala500Gly) in the coding site of C Ⅱ TA gene did not influence capability of CⅡTA trans-aetivating HLA class Ⅱgene expression.