Effectiveness and safety of robot-assisted minimally invasive transforaminal lumbar interbody fusion for degenerative lumbar spinal diseases: a systematic review and meta-analysis.

Robot-assisted (RA) technology has been widely used in spine surgery. This analysis aimed to compare the effectiveness and safety of RA minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) and fluoroscopy-assisted (FA) MIS-TLIF for degenerative lumbar spinal diseases (DLSD). PubMed, Web of Science, Cochrane Library, and China National Knowledge Infrastructure were systematically searched, and the outcomes included surgical parameters [operation time, blood loss, number of fluoroscopic, accuracy of pedicle screw position, superior facet joint violation (FJV)], and clinical indexes (Visual Analog Scale (VAS), Oswestry Disability Index (ODI), Japanese Orthopaedic Association (JOA) score, clinical efficacy, hospital stays, complications). Eleven articles involving 1066 patients were included. RA group produced better results than the FA group in operation time (WMD = - 6.59; 95% CI - 12.79 to - 0.40; P = 0.04), blood loss (WMD = - 34.81; 95% CI - 50.55 to - 19.08; P < 0.0001), number of fluoroscopic (WMD = - 18.24; 95% CI - 30.63 to - 5.85; P = 0.004), accuracy of pedicle screw position: Grade A (OR = 3.16; 95% CI 2.36-4.23; P < 0.00001), Grade B (OR = 0.39; 95% CI 0.28-0.54; P < 0.00001), Grade C (OR = 0.27; 95% CI 0.13-0.54; P = 0.0002), and Grade D (OR = 0.17; 95% CI 0.03-0.98; P = 0.05), FJV: Grade 0 (OR = 3.27; 95% CI 1.34-8.02; P = 0.010), Grade 1 (OR = 0.24; 95% CI 0.16-0.38; P < 0.00001), Grade 2 (OR = 0.24; 95% CI 0.12-0.51; P = 0.0002), and Grade 3 (OR = 0.26; 95% CI 0.07-0.93; P = 0.04). But no significant differences in VAS score, ODI, JOA score, clinical efficacy, hospital stays, and complications. These results demonstrate a significant improvement in the intraoperative course of the RA technique. However, RA-MIS-TLIF has not yet demonstrated significant advantages in terms of postoperative symptom relief and functional improvement. Future research and clinical practice should further explore the efficacy of this technique to optimize outcomes and quality of life for patients with DLSD. The study was registered in the PROSPERO (CRD42023454405).

Large-aperture electromagnetically actuated MEMS Fabry-Perot filtering chips for visible spectral imaging.

Benefitting from the inherent merits of tiny volume, customizable performance, good system compatibility and high-yield production, micro-electro-mechanical-system-based Fabry-Perot filtering chip (MEMS-FPFC) with a large aperture size gives a feasible way for the realization of miniaturized spectral imagers which can serve in many civilian and military scenarios. Although the aperture size of MEMS-FPFCs in mid-wave and long-wave infrared has reached to the centimeter scale, that of visible wavelength (VIS) MEMS-FPFC is always unsatisfied which is mainly limited by micromachining stress, especially in the thin films. In this work, we propose a large-aperture electromagnetically actuated MEMS-FPFC based on Si3N4 supporting membrane for VIS spectral imaging, which is designed with the assistance of multi-field coupling simulation model. A low-stress wafer-scale bulk micromachining process is developed to guarantee the high-quality and high-yield production for the aimed VIS MEMS-FPFCs. Finally, by the strictly controlling and rationally allocating the film stress of multi-layer film stack, VIS MEMS-FPFCs with 6 mm aperture size are thus developed, which can be tuned bidirectionally and continuously in 612-678 nm waveband with a good linear response of better than 95%. The achieved VIS MEMS-FPFCs can be utilized to construct miniaturized spectral imagers directly, aiming for such applications as intelligent agriculture, environmental protection and industrial inspection.

High expression of integrin-binding sialoprotein (IBSP) is associated with poor prognosis of osteosarcoma.

INTRODUCTION: Osteosarcoma is a malignant tumor, accounting for 20% of primary malignant bone tumors worldwide. However, the role of IBSP as a biomarker in osteosarcoma progression has not been studied yet. METHODS: 85 cases of IBSP expression and clinical characteristics were obtained from TARGET database. Through the Kaplan-Meier curve, subgroup analysis, and univariate and multivariate Cox analysis, we further assessed the independent predictive capacity of IBSP expression for overall survival (OS) and relapse-free survival (RFS). RESULTS: The mRNA expression of IBSP was higher in osteosarcoma than normal tissue (P < 0.0001). IBSP expression grouped by vital status showed statistical differences (P = 0.042). The race (P = 0.0183), vital status (P = 0.0034), and sample type (P = 0.0020) showed significant differences. IBSP expression exhibited satisfied diagnostic ability for osteosarcoma. The univariate and multivariate analysis confirmed that IBSP expression was an independent risk factor for OS (HR = 3.425, 95% CI: 1.604-7.313, P = 0.002) and RFS (HR = 3.377, 95% CI: 1.775-6.424, P < 0.001) in osteosarcoma patients. High IBSP expression was significantly associated with poor OS and RFS (P < 0.0001). The higher IBSP expression was observed in osteosarcoma (P < 0.001), confirmed by the IHC staining. The CCK-8 and colony formation assay showed that IBSP knockdown inhibits cell proliferation while overexpression promotes cell proliferation (P < 0.05). CONCLUSION: High expression of IBSP was associated with poor OS and RFS. IBSP could serve as a potential biomarker for osteosarcoma, which could aid in early detection and disease monitoring.

Co-delivery of vitamin D3 and Lkb1 siRNA by cationic lipid-assisted PEG-PLGA nanoparticles to effectively remodel the immune system in vivo.

The imbalance of the immune system can lead to the occurrence of autoimmune diseases. Controlling and regulating the proliferation and function of effector T (Teff) cells and regulatory T (Treg) cells becomes the key to treating these diseases. Dendritic cells (DCs), as dedicated antigen-presenting cells, play a key role in inducing the differentiation of naive CD4+ T cells. In this study, we designed a cationic lipid-assisted PEG-PLGA nanoparticle (NPs/VD3/siLkb1) to deliver 1,25-dihydroxyvitamin D3 (VD3) and small interfering RNA (siRNA) to DC cells in the draining lymph nodes. By modulating the phenotypic changes of DC cells, this approach expands Treg cells and reduces the occurrence of autoimmune diseases. Thus, this study provides a novel approach to alleviating the occurrence and development of autoimmune diseases while also minimizing the risk of unwanted complications.

Factors Affecting Psychological and Health-Related Quality-of-Life Status in Children and Adolescents with Congenital Heart Diseases.

Congenital heart disease (CHD), a severe cardiac defect in children, has unclear influences on young patients. We aimed to find the impacts of differently structure heart defects and various treatments on psychology and health-related quality of life (HRQoL) in CHD children and adolescents. CHD patients aged between 6 and 18 years old visited our hospital from 1 May 2018 to 31 September 2018, and their principal caregivers were asked to participate. We used two validated questionnaires, Children Depression Inventory-TW (CDI-TW) and Child Health Questionnaire-Parent Form 50 (CHQ-PF 50), to evaluate CHD patients' psychological and HRQoL conditions. Participants were grouped based on their cardiac defects and previous treatments. We analyzed the results via summary independent-samples t-test with post hoc Bonferroni correction and multivariant analysis. Two hundred and seventy-seven children and their principal caregivers were involved. There was no apparent depressive condition in any group. Single cardiac defect patients exhibited similar HRQoL to controls; simultaneously, those with cyanotic heart disease (CyHD), most multiple/complex CHDs children and adolescents, and those who received invasive treatments had poorer HRQoL. CyHD impacted the most on patients' psychological and HRQoL status. Patients with sole cardiac defect could live near-normal lifes; on the other hand, CyHD had the worst effects on patients' psychology and HRQoL.

Potential application of hydrogel to the diagnosis and treatment of multiple sclerosis.

Multiple sclerosis (MS) is a chronic demyelinating disease of the central nervous system. This disorder may cause progressive and permanent impairment, placing significant physical and psychological strain on sufferers. Each progress in MS therapy marks a significant advancement in neurological research. Hydrogels can serve as a scaffold with high water content, high expansibility, and biocompatibility to improve MS cell proliferation in vitro and therapeutic drug delivery to cells in vivo. Hydrogels may also be utilized as biosensors to detect MS-related proteins. Recent research has employed hydrogels as an adjuvant imaging agent in immunohistochemistry assays. Following an overview of the development and use of hydrogels in MS diagnostic and therapy, this review discussed hydrogel's advantages and future opportunities in the diagnosis and treatment of MS.

Predictive risk factors for recollapse of cemented vertebrae after percutaneous vertebroplasty: A meta-analysis.

BACKGROUND: As one of the most common complications of osteoporosis, osteoporotic vertebral compression fracture (OVCF) increases the risk of disability and mortality in elderly patients. Percutaneous vertebroplasty (PVP) is considered to be an effective, safe, and minimally invasive treatment for OVCFs. The recollapse of cemented vertebrae is one of the serious complications of PVP. However, the risk factors associated with recollapse after PVP remain controversial. AIM: To identify risk factors for the recollapse of cemented vertebrae after PVP in patients with OVCFs. METHODS: A systematic search in EMBASE, MEDLINE, the Cochrane Library, and PubMed was conducted for relevant studies from inception until March 2020. Studies investigating risk factors for the recollapse of cemented vertebrae after PVP without additional trauma were selected for analysis. Odds ratios (ORs) or standardized mean differences with 95% confidence interval (CI) were calculated and heterogeneity was assessed by both the chi-squared test and the I-squared test. The methodological quality of the included studies was assessed according to the Newcastle-Ottawa Scale. RESULTS: A total of nine case-control studies were included in our meta-analysis comprising 300 cases and 2674 controls. The significant risk factors for the recollapse of cemented vertebrae after PVP in OVCF patients were fractures located at the thoracolumbar junction (OR = 2.09; 95%CI: 1.30 to 3.38; P = 0.002), preoperative intravertebral cleft (OR = 2.97; 95%CI: 1.93 to 4.57; P < 0.00001), and solid lump distribution pattern of the cement (OR = 3.11; 95%CI: 1.91 to 5.07; P < 0.00001). The analysis did not support that age, gender, lumbar bone mineral density, preoperative visual analogue scale score, injected cement volume, intradiscal cement leakage, or vertebral height restoration could increase the risk for cemented vertebra recollapse after PVP in OVCFs. CONCLUSION: This meta-analysis suggests that thoracolumbar junction fractures, preoperative intravertebral cleft, and solid lump cement distribution pattern are associated with the recollapse of cemented vertebrae after PVP in OVCF patients.

Antagonism of GPR4 with NE 52-QQ57 and the Suppression of AGE-Induced Degradation of Type II Collagen in Human Chondrocytes.

Osteoarthritis (OA) is a common degenerative joint disease for which an effective therapeutic strategy has not yet been established. AGEs are widely recognized as a contributor to OA pathogenesis. GPR4, a recently discovered proton-sensing transmembrane receptor, has been shown to possess a wide range of physiological functions. However, the potential role of this receptor in chondrocytes and the pathogenesis of OA is unclear. In the present study, we investigated the potential of GPR4 to modulate the effects of advanced glycation end products (AGEs) in SW1353 human chondrocytes. First, we demonstrate that GPR4 is fairly expressed in SW1353 chondrocytes and that exposure to AGEs increases the expression of this transmembrane receptor. Second, we found that antagonism of GPR4 with NE 52-QQ57 significantly inhibited the AGE-induced increased expression of several key inflammatory cytokines and signaling molecules, including tumor necrosis factor-α (TNF-α), interleukin (IL)-1ß, IL-6, inducible nitric oxide synthase (iNOS), nitric oxide (NO), cyclooxygenase 2 (COX2), and prostaglandin E2 (PGE2). We also found that antagonisn of GPR4 had a remarkable ability to rescue type II collagen from AGE-induced degradation by inhibiting the expression of matrix metalloproteinase (MMP)-3 and MMP-13. As a key pro-inflammatory signaling pathway, we further tested the effect of GPR4 antagonism on the activation of nuclear factor-κB (NF-κB) and found that NF-κB activation was indeed suppressed, thereby indicating that the NF-κB signaling pathway may mediate the effects of GPR4 antagonism described above. These findings provide a basis for further research into the role of GPR4 -mediated signaling in OA.

Long non-coding RNA SNHG16 reduces hydrogen peroxide-induced cell injury in PC-12 cells by up-regulating microRNA-423-5p.

Functions of long non-coding RNAs (lncRNAs) have been widely probed in spinal cord injury (SCI). But, the influences of lncRNA-small nucleolar RNA host gene 16 (lncRNA-SNHG16) is still not well documented in SCI. The study explored the impacts of SNHG16 on H2O2-injured PC-12 cells. PC-12 cells were disposed with H2O2, cell viability, apoptosis, autophagy and ROS level were detected. RT-qPCR was executed to explore SNHG16 or miR-423-5p expression in H2O2-stimulated cells. After transfection with pc-SNHG16 or miR-423-5p inhibitor, the functions of SNHG16 and miR-423-5p in H2O2-injured cells were studied. AMPK and ERK1/2 pathways were finally assessed by western blot. We found that H2O2 evoked cell injury in PC-12 cells, and repressed SNHG16 was observed in H2O2-disposed cells. Overexpressed SNHG16 prominently alleviated H2O2-induced cell injury as indicated by repressing cell apoptosis, autophagy and ROS level. Additionally, SNGH16 enhanced miR-423-5p expression, and miR-423-5p inhibition abrogated the protective effect of SNGH16 on H2O2-injured PC-12 cells. SNGH16 mediated AMPK and ERK1/2 pathways via up-regulating miR-423-5p in H2O2-injured PC-12 cells. In conclusion, these findings indicated that SNGH16 reduced H2O2-evoked cell injury by mediating miR-423-5p in PC-12 cells. The findings might uncover the effect of SNHG16 on SCI, which provide a new reference for remedying SCI. Highlights H2O2 evokes cell injury in PC-12 cells; SNHG16 reduces H2O2-induced cell injury in PC-12 cells; SNGH16 enhances miR-423-5p expression in H2O2-stimulated PC-12 cells; MiR-423-5p inhibition abrogates the protective effect of SNGH16 in PC-12 cells; SNGH16 mediates AMPK and ERK1/2 pathways by up-regulating miR-423-5p.

microRNA-203 promotes proliferation, differentiation, and migration of osteoblasts by upregulation of Msh homeobox 2.

Despite the improvements in fracture healing, about 10% of patients undergo abnormal healing. As a tumor suppressor, upregulation of microRNA (miR)-203 has been observed in osteogenic differentiation. Herein, we aimed to explore the functional role of miR-203 in osteoblasts as well as the underlying mechanisms. The expression of miR-203 in MC3T3-E1 cells that underwent osteogenic differentiation was determined by quantitative reverse transcription PCR (qRT-PCR). The effects of aberrantly expressed miR-203 on cell viability, migration, and expressions of proteins associated with proliferation, migration, and osteogenic differentiation were measured by using a Cell Counting Kit-8 assay, Transwell cell migration assay, and western blot/qRT-PCR, respectively. The possible downstream factor of miR-203 was subsequently studied. Finally, involvements of the mitogen-activated protein kinase (MAPK)/activator of transcription (STAT) pathways were assessed by western blot. We found that the miR-203 level was increased in osteogenic differentiation of MC3T3-E1 cells with increasing duration time (28th day, p < 0.001). After cell transfection, we interestingly found that miR-203 overexpression could increase cell viability (p < 0.05), promote proliferation, migration (p < 0.05), and osteogenic differentiation, and upregulate Msh homeobox 2 (Msx2) expression. Furthermore, Msx2 knockdown was proved to abrogate the effects of miR-203 overexpression on MC3T3-E1 cells. Finally, phosphorylated levels of key kinases in the MAPK/STAT pathways were increased by miR-203 overexpression via upregulating Msx2 expression. In conclusion, miR-203 overexpression promoted proliferation, migration, and osteogenic differentiation of MC3T3-E1 cells through upregulating Msx2 along with activation of the MAPK/STAT pathways.

Serotype distribution and clinical correlation of Streptococcus agalactiae causing invasive disease in infants and children in Taiwan.

BACKGROUND: Streptococcus agalactiae, or group B Streptococcus (GBS), remains to be one of the leading pathogens causing invasive infections in infants. METHODS: The clinical GBS isolates from sterile sites of patients younger than 18 years old were collected from October 1998 to December 2014 in two hospitals in Taiwan. Medical records were retrospectively reviewed. Every isolate was serotyped with a multiplex PCR assay. Multilocus sequence typing (MLST) was performed in representative isolates of different serotypes. A total of 205 GBS isolates were collected from 181 patients with 182 infection episodes. RESULTS: Serotype Ia was the most common in patients less than 72 h old, whereas III the most common in patients older than 72 h. In early-onset disease (0-6 days), Ia and III each caused 27.5% of the infection, followed by Ib (14.5%). In late-onset disease (7-89 days), serotype III predominated (75.3%), followed by Ia (10.1%) and Ib (6.8%). Thirty-one episodes (17%) were complicated with culture-confirmed meningitis. We compared serotype Ia and III patients, and found that serotype Ia patients were significantly younger (median age, 3 days), had more perinatal maternal fever and higher mortality. ST17 and ST19 were exclusively found in serotype III, while ST23 and ST24 comprised of 85% of serotype Ia. CONCLUSION: In Taiwan, serotypes Ia and III are the most common cause for early-onset and late-onset neonatal GBS infections, respectively. Some differences in the clinical features of invasive GBS infections caused by serotype Ia and III were observed.

Functional and biological studies of α-galactosidase A variants with uncertain significance from newborn screening in Taiwan.

Fabry disease is an X-linked disorder resulted from deficiency of α-galactosidase A (GLA) activity. In Taiwan, a total of 792,247 newborns were screened from 2008 to 2014 in two newborn screening centers, and 13 variants of uncertain significance (VOUS) in the GLA gene were identified. To determine whether these variants were pathogenic or not, functional, biochemical, clinical and pedigree analyses were performed. In vitro functional assay was established through site-directed mutagenesis, and four in silico tools were used to predict pathogenesis. The enzyme activity of dried blood spots and plasma metabolite lyso-Gb3 level from subjects with the variants were measured. Additionally, clinical manifestations were evaluated extensively from the subjects and their relatives. Our results revealed that p.G104V, p.I232T, p.D322H, and p.G360C all exhibited relatively low residual enzyme activities and elevated plasma lyso-Gb3 level. These data strongly suggest that these Fabry mutations may cause classical or later-onset phenotypes. In contrast, neither significantly clinical symptoms nor elevated lyso-Gb3 level was found in cases with p.P60S, p.A108T, p.S304T, p.R356Q, and p.P362T variants, which may be non-pathogenic or milder forms of Fabry variants. More data need to be included for the patients with p.N53D, p.P210S, p.M296L, and p.K391T variants. The established system provides us more information to classify these GLA variants.

Cervical Disc Arthroplasty Versus Anterior Cervical Discectomy and Fusion for Incidence of Symptomatic Adjacent Segment Disease: A Meta-Analysis of Prospective Randomized Controlled Trials.

STUDY DESIGN: Meta-analysis of randomized controlled trials. OBJECTIVE: To evaluate the reported rate of adjacent segment disease (ASD) of cervical disc arthroplasty (CDA) compared with anterior cervical discectomy and fusion (ACDF). SUMMARY OF BACKGROUND DATA: Motion-maintaining technologies such as CDA have developed rapidly because of the concern of ASD. Till date, however, it still has been under debate whether CDA is superior to ACDF regarding the incidence of ASD. METHODS: We comprehensively searched PubMed, EMBASE, and Cochrane Central Register of Controlled Trails for prospective randomized controlled trials (RCTs) that reported the incidence of ASD between CDA and ACDF. The retrieved results were last updated on November 20, 2015 without language restrictions. Two independent authors selected qualified studies, assessed methodological quality, and extracted requisite data. RESULTS: Fourteen relevant RCTs involving 3235 individuals with a follow-up period of 2 to 7 years were included in the meta-analysis (1696 in CDA group and 1539 in ACDF group). The outcomes indicated that CDA was superior to ACDF considering the lower rate of ASD (risk ratio, 0.57; 95% confidence interval, 0.37 to 0.87; P = 0.009). And compared with ACDF, there were significantly fewer adjacent segment reoperations in the CDA group (risk ratio, 0.47; confidence interval, 0.32 to 0.70; P = 0.0002). Subgroup analysis stratified by different types of disc prostheses was also performed. CONCLUSION: CDA was superior to ACDF regarding fewer ASDs and relative reoperations on the basis of available evidence from a meta-analysis of 14 RCTs. CDA may be a better surgical procedure to reduce the incidence of ASD for patients with cervical disc disease compared with ACDF. Further well-designed studies should continue to pay attention to excellent patients with longer-term follow-up to evaluate the incidence of ASD of these two procedures. LEVEL OF EVIDENCE: 1.

Clinical observations and treatment of pediatric homozygous familial hypercholesterolemia due to a low-density lipoprotein receptor defect.

BACKGROUND: Clinical observation and treatment of children with homozygous familial hypercholesterolemia (HoFH) has rarely been reported. We report clinical observations and treatment of 10 ethnic Chinese children with HoFH due to low-density lipoprotein receptor (LDLR) defect. OBJECTIVES: In children with HoFH, we evaluated the response to conventional cholesterol-lowering drug therapy and performed LDLR gene analysis. METHODS: A retrospective review of lipid profile changes in pediatric patients diagnosed with HoFH seen in our pediatric endocrinology outpatient clinic was performed. HoFH was diagnosed by molecular study of these patients and their parents. RESULTS: One novel (c.64del G) and 12 known mutations were found in the LDLR gene. Mutation of p.C308Y was the most common and was found in 26% of the studied alleles.Seven patients had fair responses to conventional drug therapy (high-dose statin with ezetimibe) with a reduction of 50% or more of the total cholesterol levels. The low-density lipoprotein-cholesterol levels of three patients decreased to lower than 160 mg/dL. One who had a good response to conventional drug therapy developed significant atheromatous plaques (largest plaque: 7.4 × 2.7 cm) in the extracranial carotid arteries and myocardial ischemia changes at 11 years old. CONCLUSION: The results suggest that despite aggressive therapy, many patients are not well controlled; atherosclerosis may progress, and novel therapies are required.

Age at First Cardiac Symptoms in Fabry Disease: Association with a Chinese Hotspot Fabry Mutation (IVS4+919G>A), Classical Fabry Mutations, and Sex in a Taiwanese Population from the Fabry Outcome Survey (FOS).

This is a descriptive analysis of a cohort of 59 Taiwanese patients with Fabry disease and either classical Fabry or cardiac variant IVS4+919G>A (IVS4) mutations from a disease registry, the Fabry Outcome Survey (FOS; sponsored by Shire). Most of our classical Fabry patients were symptomatic and were identified upon seeking medical advice at our clinics, whereas most of our IVS4 patients attended our clinics after newborn screening identified this mutation in their grandsons. The objective was to determine differences in cardiac manifestations between patients with classical Fabry or IVS4 mutations by comparing age at onset of selected cardiac symptoms. Data were extracted in August 2013 and analyzed retrospectively. Fifty-nine Taiwanese patients (median age at extract 60.7 years [range 15.0-86.9]; n = 36 [61%] male) with proven IVS4 (n = 41 [69%]) or classical Fabry mutations (n = 18 [31%]) had available data on cardiac symptoms. Of 55 (93%) patients with reported left ventricular hypertrophy (LVH), mean [SD] age (years) at first symptom was lower in classical Fabry males (30.0 [15.1]; n = 4) than classical Fabry females (49.6 [8.9]; n = 11; p < 0.05), but not in IVS4 females (57.4 [13.7]; n = 10) compared with IVS4 males (55.9 [11.3]; n = 30). Mean age at first LVH diagnosis was significantly lower in classical Fabry males versus IVS4 males (p < 0.05). No significant difference in age at onset of arrhythmia or conductive abnormality, chest pain, or palpitations or cardiac syncope was found between the groups. The most noteworthy finding of this study is the lack of a significant gender sex difference in age at onset of cardiac symptoms in IVS4 patients.

Globotriaosylsphingosine (lyso-Gb3) might not be a reliable marker for monitoring the long-term therapeutic outcomes of enzyme replacement therapy for late-onset Fabry patients with the Chinese hotspot mutation (IVS4+919G>A).

BACKGROUND: In Taiwan, DNA-based newborn screening showed a surprisingly high incidence (1/875 in males and 1/399 in females) of a cardiac Fabry mutation (IVS4 + 919G > A). However, the natural course, long-term treatment outcomes and suitable biomarkers for monitoring the therapeutic outcomes of these patients are largely unknown. METHODS: Fabry disease (FD) patients who had received enzyme replacement therapy (ERT) for more than 1 year were enrolled in this study from December 2008 to April 2013. Periodic echocardiography and serum globotriaosylsphingosine (lyso-Gb3) analysis were carried out. Before and after ERT, left ventricular mass index (LVMI) and serum lyso-Gb3 level were compared and the correlation between the change of LVMI and the change of serum lyso-Gb3 were also analyzed. RESULTS: Thirty-six patients, in four patient groups, were enrolled: (1) 16 males with IVS4 + 919G > A mutation; (2) 7 females with IVS4 + 919G > A mutation; (3) 2 males with classical mutations; and (4) 11 females with classical mutations. The follow-up period was 13-46 months. There were significant LVMI reductions after ERT in all four groups after excluding confounding factors. However, interestingly, serum lyso-Gb3 decreased significantly in the early period after ERT in all groups, but increased gradually after an average of 11.1 months after ERT in late-onset male and female Fabry groups, even when their LVMI still decreased or remained stable. Furthermore, there was no correlation between the change of serum lyso-Gb3 and the change of LVMI in both classical and IVS4 + 919G > A FD patients. CONCLUSION: Although lyso-Gb3 has a high diagnostic sensitivity in late-onset Fabry patients and has a good response to ERT during the early stages, it might not be a reliable marker for monitoring the long-term therapeutic outcomes of ERT for late-onset Fabry patients with the Chinese hotspot mutation (IVS4 + 919G > A).

Endomyocardial biopsies in patients with left ventricular hypertrophy and a common Chinese later-onset Fabry mutation (IVS4 + 919G > A).

BACKGROUND: In Taiwan, DNA-based newborn screening showed a surprisingly high incidence of a cardiac Fabry mutation (IVS4 + 919G > A). The prevalence of this mutation is too high to be believed that it is a real pathogenic mutation. The purpose of this study is to identify the cardiac pathologic characteristics in patients with left ventricular hypertrophy and this mutation METHODS AND RESULTS: Endomyocardial biopsies were obtained in 22 patients (Median age: 61, males: 17; females: 5) with left ventricular hypertrophy and the IVS4 + 919G > A mutation; five patients had not received enzyme replacement therapy (ERT) before biopsy, while the other 17 patients had received ERT from 8 months to 51 months. Except for three patients who had received ERT for more than 3 years, all other patients showed significant pathological change and globotriaosylceramide (Gb3) accumulation in their cardiomyocytes. In contrast to classical Fabry patients, no Gb3 accumulation was found in the capillary endothelial cells of any of our patients. Fourteen patients (63.6%) were found to have myofibrillolysis. CONCLUSIONS: All of the untreated and most of the treated IVS4 + 919G > A patients showed typical pathological changes of Fabry disease in their cardiomyocytes. No endothelial accumulation of Gb3 was found, which is similar to the findings of several previous reports regarding later-onset Fabry disease. This result highly suggests that the IVS4 + 919G > A is a real pathogenic later-onset Fabry mutation.

High-throughput detection of common sequence variations of Fabry disease in Taiwan using DNA mass spectrometry.

BACKGROUND: In view of the therapeutic benefits resulting from early intervention for Fabry disease, our team has implemented an enzyme-based newborn screening in Taiwan since 2008. However, we found that most heterozygous females cannot be detected. To improve the screening efficiency, a more effective method for GLA gene genotyping is necessary. METHODS: As the suspected mutations are limited to only 29 different spots in Taiwanese, a panel of Sequenom iPLEX assay was designed for rapid screening of GLA variations. To determine the accuracy and sensitivity of this assay, previously diagnosed and undiagnosed DNA samples were analyzed by this genotyping assay and Sanger sequencing. In addition, DNA extracted from dried blood spots was also tested. RESULTS: Sequenom iPLEX assay is accurate and cost-effective, identifying the sequence variations, which were designated in the panel. It identified common GLA variants in DNA samples extracted from whole blood or dried blood spots with 100% accuracy and sensitivity. CONCLUSIONS: Sequenom iPLEX assay is suitable for Fabry newborn screening when hotspot mutations and common variations are known in a well-studied population. In addition, this assay can also be applied for first-line determination of GLA variant sequences in suspected subjects of high-risk patients, or newborns.

Detecting multiple lysosomal storage diseases by tandem mass spectrometry--a national newborn screening program in Taiwan.

BACKGROUND: Interest in lysosomal storage diseases in newborn screening programs has increased in recent years. Two techniques, fluorescence (4-MU) and tandem mass spectrometry (MS/MS) methods are frequently used. We report a pilot study of large scale newborn screening for Fabry, Pompe, Gaucher, and MPS I diseases by using the MS/MS method in Taiwan and compared the performance of the MS/MS with 4-MU methods. METHODS: More than 100,000 dried blood spots (DBSs) were collected consecutively as part of the national Taiwan newborn screening programs. The enzyme activities were detected by the MS/MS method from a DBS punch. Mutation analysis was further performed for newborns with detected enzyme deficiency. RESULTS: The DNA sequence analysis for suspected cases revealed 64 newborns with confirmed Fabry mutations, 16 were classified as infantile or late-onset Pompe disease, and 1 was characterized as Gaucher disease. The positive predict value increased from 4.0% to 7.1% in the Pompe study, and from 61.0% to 95.5% in the Fabry study by the MS/MS method compared to 4-MU assay. CONCLUSIONS: The MS/MS method has been validated as a more specific, powerful and efficient tool than the 4-MU assay. It also provided a multiplex solution of newborn screening for lysosomal storage diseases.