[Child with sitosterolemia initially presenting with hemolytic anemia and thrombocytopenia: a case repore and literrature review].

This article focuses on a case study of sitosterolemia in a child who initially presented with hemolytic anemia and thrombocytopenia. Sitosterolemia is a rare autosomal recessive lipid metabolism disorder, difficult to diagnose due to its non-typical clinical manifestations. The 8-year-old patient was initially misdiagnosed with pyruvate kinase deficiency. Comprehensive biochemical and molecular biology analyses, including gene sequencing, eventually led to the correct diagnosis of sitosterolemia. This case highlights the complexity and diagnostic challenges of sitosterolemia, emphasizing the need for increased awareness and accurate diagnosis in patients presenting with similar symptoms.

Dexmedetomidine represses proliferation and promotes apoptosis of esophageal cancer cells by regulating C-Myc gene expression via the ERK signaling pathway.

OBJECTIVE: The aim of this study was to investigate the effects of dexmedetomidine (DEX) on proliferation and apoptosis of esophageal cancer (EC) cells, and to explore the possible underlying mechanism. MATERIALS AND METHODS: EC cells (Eca109) were randomly divided into two groups, namely, Control group and DEX group. The viability, proliferation, and apoptosis of Eca109 cells were detected using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 5-Ethynyl-2'-deoxyuridine (EdU) staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining, respectively. Meanwhile, the messenger ribonucleic acid (mRNA) and protein expression levels of extracellular signal-regulated kinase (ERK) 1/2 and c-Myc in Eca109 cells were measured by quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) and Western blotting, respectively. RESULTS: The viability of Eca109 cells was remarkably weakened in DEX group when compared with Control group (p<0.05). DEX could significantly inhibit the proliferation and promote the apoptosis of Eca109 cells (p<0.05). Moreover, the mRNA and protein levels of ERK1/2 and c-Myc in Eca109 cells declined notably (p<0.05). CONCLUSIONS: DEX represses the proliferation and facilitates the apoptosis of Eca109 cells prominently. The possible underlying mechanism may be associated with the inhibition of c-Myc gene expression through the ERK signaling pathway.

[Platelet derived growth factor-BB regulates phenotype transformation of pulmonary artery smooth muscle cells via SIRT3 affecting glycolytic pathway].

Objective: To investigate whether platelet-derived growth factor-BB (PDGF-BB) can regulate phenotypic transformation of pulmonary artery smooth muscle cells (PASMCs) via SIRT3 affecting glycolytic pathway. Methods: The PASMCs were isolated from Sprague Dawley rats. PASMCs were divided into 3 groups by using 2-deoxyglucose (2-DG), an inhibitor of the glycolytic pathway: normal control group, PDGF-BB group(30 ng/ml) and PDGF-BB (30 ng/ml)+2-DG (10 mmol/L) group. In lentivirus-mediated overexpression assay, cells were divided into control group, PDGF-BB group(30 ng/ml), PDGF-BB+deacetylase sirtuin-3 (SIRT3) overexpression group and PDGF-BB+empty vector group. The expression levels of phenotype related index such as α-smooth muscle actin (α-SMA), smooth muscle myosin heavy chain (SM-MHC), calponin, vimentin were detected by qRT-PCR and Western blot. Meanwhile, the expression of α-SMA was detected by cellular immunofluorescence staining. EDU staining was used to detect the proliferation of PASMCs. The expression of SIRT3 was detected by Western blot. The expressions of glucose transporter 1 and aerobic glycolytic enzymes were detected by qRT-PCR and Western blot in lentivirus-mediated overexpression assay. Results: (1) PDGF-BB affects PASMCs phenotypic transformation through glycolytic pathway: compared with normal control group, PDGF-BB significantly decreased the expressions of contractile phenotype markers such as α-SMA, SM-MHC, calponin mRNA and protein (all P<0.05), but it increased the expressions of the synthetic phenotype marker vimentin mRNA and protein (both P<0.05). Cellular immunofluorescence assay showed that PDGF-BB significantly decreased the number of α-SMA positive cells, while 2-DG reversed the process. (2) PDGF-BB promoted cell proliferation through glycolytic pathway: the proliferation of PASMCs was significantly higher in PDGF-BB group than in control group (P<0.05), and which could be significantly reduced by 2-DG (P<0.05). (3) PDGF-BB inhibited the expression of SIRT3 protein in PASMCs: the expression of SIRT3 protein in PDGF-BB group was lower than that in control group (P<0.05). (4) PDGF-BB affected glycolytic pathway through SIRT3:compared with the control group, PDGF-BB significantly increased the expression levels of glucose transporter 1 (Glut1), hexokinase 2 (HK2) and 6-phosphfructo-2-kinase 3 (PFKFB3) mRNA (all P<0.05), which was reserved by over-expression of SIRT3. There were no significant difference in mRNA expression levels between PDGF-BB group and PDGF-BB+empty vector group (P>0.05).Compared with the control group, PDGF-BB significantly increased the expression levels of Glut1, HK2 and PFKFB3 protein(all P<0.05), which was reserved by over-expression of SIRT3. There were no significant differences in protein expression levels between PDGF-BB group and PDGF-BB+empty vector group (all P>0.05). Conclusion: PDGF-BB regulates phenotypic transformation of PASMCs via SIRT3 affecting glycolytic pathway.

[Awareness of preconceptional care and its related factors in women of child-bearing age with type 1 diabetes].

Objective: To investigate the awareness of preconception care among women of child-bearing age with type 1 diabetes (T1DM) and their self-management status, in order to provide evidence for establishment of management pathway for women with T1DM in pregnancy in China. Methods: This cross-sectional survey recruited female participants of child-bearing age from the cohort of Guangdong Type 1 Diabetes Translational Medicine Study conducted between June 2011 and December 2017. The participants were asked to fill out a questionnaire on the awareness of preconception care, their frequency of self-monitoring of blood glucose (SMBG) and other related variables. Chi-squared test or chi-squared test for trend was used in comparisons of categorical variables, and logistic regression analysis was performed to assess associated factors. Results: Totally, 441 women of child-bearing age with T1DM were investigated. The results show that their awareness of preconception care was poor (15.42%, 68/441). Higher educational level (χ(2trend)=3.990, P=0.046), experience of post-diabetes education evaluation (P<0.001), and better coverage of different modules in diabetes education (survival skills: χ(2)=7.525, P=0.004; basic knowledge: χ(2)=8.598, P=0.002; advanced knowledge: P<0.001) were associated with better awareness of preconception care. The average frequency of SMBG in these participants was 0.29 (0.14, 2.00) times per day, and only 8.5% (37/435) of them reached the frequency (≥4 times per day) recommended by guidelines. Moreover, 21.1% (92/435) of them hardly ever performed SMBG. Conclusion: Child-bearing age women with T1DM in Gunangdong had poor awareness of preconception care, with a much lower SMBG frequency than recommendation.

[Glycemic control and its associated factors in children and adolescents with type 1 diabetes mellitus].

Objective: To explore the factors associated with glycemic control in children and adolescents with type 1 diabetes mellitus (T1DM) treated with continuous subcutaneous insulin infusion (CSII). Methods: Subjects were enrolled from the Guangdong Type 1 Diabetes Translational Medicine Study between June 2011 and August 2017. Patients with T1DM aged less than 18 years and treated with CSII for at least 6 months were included. Demographic data and clinical information on self-monitoring of blood glucose (SMBG), glycosylated hemoglobin (HbA1c) and insulin treatment were collected. Participants were categorized based on HbA1c levels as sufficient control group (HbA1c<7.5% ) and insufficient control group ( HbA1c≥7.5%). A multivariate logistic regression model was used to examine the factors associated with glycemic control. Results: A total of 142 participants (76 females, 66 males) with a median age of 13.0 (9.9, 15.0) years and a median disease duration of 3.0 (1.6, 5.0) years were enrolled. HbA1c was (8.2±2.0)% and 41.55%(59/142) of patients achieved the target for HbA1c. The frequency of SMBG was 5.0 (2.0, 8.0) and 3.0 (1.0, 4.0) tests per day (P<0.001), and the frequency of hypoglycemia was 2.0 (0.8, 4.0) and 1.0 (0, 2.0) times per week (P=0.003) in sufficient control group and insufficient control group, respectively. Sufficient glycemic control (HbA1c <7.5%) was associated with the frequency of SMBG (OR=1.238, 95% CI: 1.088-1.409, P=0.001). Conclusion: A higher frequency of SMBG is one of the key factors to achieve sufficient glycemic control among children and adolescents with T1DM treated with CSII.

[Insulin dose analysis during pregnancy in type 1 diabetic patients treated with insulin pump therapy].

Objective: To analyze the insulin dose of type 1 diabetic patients who treated with insulin pump therapy during pregnancy in order to explore the features of these patients' insulin requirement during gestation. Methods: A total of 12 well-controlled type 1 diabetic women patients who were treated with insulin pump therapy before and during gestation without any adverse pregnancy outcomes from June 2011 to December 2014 were selected from Guangdong Type 1 Diabetes Translational Medicine Study and included in the study. Demographic data, hemoglobin A1c (HbA1c) before pregnancy and before delivery, insulin dose, hypoglycemia episodes and pregnancy outcomes were collected to analyze the insulin dose of preconception, the 1(st,) the 2(nd) and the 3(rd) trimester to analyze the requirement of insulin before and throughout pregnancy. Results: Subjects were (26.9±2.6) years old, with a diabetes duration of (6.6±4.4) years. HbA1c were (5.8±0.5)% before conception. The preconception total daily insulin dose, basal rate, bolus and bolus proportion were (0.60±0.18)U/kg, (0.28±0.10)U/kg, (0.32±0.13)U/kg and (54.8±12.9)%, respectively. Both of the insulin dose indexes mentioned above changed significantly in different trimesters compared with that in preconception (P value was <0.001, 0.034, <0.001 and <0.001, respectively). The total daily insulin dose, bolus and bolus proportion kept increasing during pregnancy. In the 1(st,) the 2(nd) and the 3(rd) trimester, the total daily insulin dose rose by 0.2%, 45.4% and 72.7%, respectively, the bolus rose by 8.0%, 72.2% and 106.8%, respectively, and the bolus proportion rose by 8.0%, 16.8% and 19.0%, respectively. While the basal rate decreased by 9.0% in the 1(st) trimester and rose by 14.1% and 32.9% in the 2(nd) and 3(rd) trimester, respectively. Conclusions: In well-controlled pregnant women with type 1 diabetes mellitus, insulin requirement increased throughout pregnancy. Most of the increased insulin requirement was attributed to the bolus instead of the basal rate. When titrating the dose of insulin for the pregnant women complicated with type 1 diabetes mellitus, the physicians should consider their features of insulin requirement so as to optimize the glycemic control.

[Insulin regimes and impact on glycemic control in patients with type 1 diabetes].

Objective: To describe the insulin regimens and their associations with glycemic control and to explore factors associated with intensive insulin therapy. Methods: Patients with type 1 diabetes (T1DM) were recruited from Guangdong Type 1 Diabetes Mellitus Translational Medicine Study which was conducted in 16 centers in Guangdong province. The demographic and clinical data were collected. Patients were grouped according to different insulin regimens: insulin pump (R1), basal insulin plus regular insulin or short-acting insulin (R2), insulin injection 1-3 times per day (R3). Distribution of insulin regimens and the relationships between insulin regimens and hemoglobin A1c (HbA1c) were described. Multivariate logistic regression was used to identify factors associated with intensive insulin therapy. Results: A total of 1 421 patients with the age of 27.8 (19.4, 38.3) years and a duration of T1DM of 3.3 (0.5, 7.1) years were recruited. There was 12.3% of patients in R1 (n=175), 35.5% in R2 (n=504), and 52.2% in R3 (n=742), respectively. HbA1c was 8.0 (6.8, 9.3)%, 8.9 (7.1, 11.8)%, and 9.2 (7.5, 11.4)% in R1, R2, R3, respectively, and it was associated with insulin regimens (P<0.001). HbA1c target rate was 32.3%, 21.1%, 17.8% in R1, R2, R3, respectively (P=0.002). Older age (OR=1.01, P=0.027), higher education level (college or above) (OR=1.56, P=0.003), and higher household income (>30 000 yuan per year per person)(OR=1.45, P=0.009) were associated with intensive insulin therapy in adult patients. Conclusions: The study suggested that insulin therapy need to be optimized in patients with T1DM. The optimization of insulin regimens and diabetes education may be helpful for improvement of glycemic control.

[Preliminary safety and stability assessment of orthodontic miniscrew implantation guided by surgical template based on cone-beam CT images].

OBJECTIVE: To develop surgical templates for orthodontic miniscrew implantation based on cone-beam CT(CBCT)three-dimensional(3D)images and to evaluate the safety and stability of implantation guided by the templates. METHODS: DICOM data obtained in patients who had CBCT scans taken were processed using Mimics software, and 3D images of teeth and maxillary bone were acquired. Meanwhile, 3D images of miniscrews were acquired using Solidworks software and processed with Mimics software. Virtual position of miniscrews was determined based on 3D images of teeth, bone, and miniscrews. 3D virtual templates were designed according to the virtual implantation plans. STL files were output and the real templates were fabricated with stereolithographic appliance(SLA). Postoperative CBCT scans were used to evaluate the implantation safety and the stability of miniscrews were investigated. RESULTS: All the templates were positioned accurately and kept stable throughout the implantation process. No root damage was found. The deviations were(1.73±0.65)mm at the corona, and(1.28±0.82)mm at the apex, respectively. The stability of miniscrews was fairly well. CONCLUSIONS: Surgical templates for miniscrew implantation could be acquired based on 3D CBCT images and fabricated with SLA. Implantation guided by these templates was safe and stable.

Comparative RNA profile analysis of idiopathic dilated cardiomyopathy and ischemic cardiomyopathy.

Previous research has focused on revealing the functions of each individual gene and/or pathway in idiopathic dilated cardiomyopathy (DCM) or ischemic cardiomyopathy (IC). However, the common or specific pathways of the initiation and processes of DCM and IC are still unclear. Here, we attempted to uncover the critical genes and potential molecular networks that play important roles in DCM and IC progression commonly or specifically. The transcriptional profiles from normal and DCM or IC patient samples were analyzed and compared using bioinformatic methods. Initially, the normal and DCM or IC sample data were processed and the most notable differentially expressed genes (DEGs) from DCM or IC were identified. By comparing the DEGs from DCM with those from IC, the DCM- and IC-specific DEGs were identified. The gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses indicated the significance of multiple biological processes as well as signaling pathways that affect heart function and DCM or IC progression. Protein-protein interaction network analysis identified the relationships between different genes, and some important genes such as MYC and FN1 were found to be hubs, which master each individual module of DCM-specific and IC-specific DEGs, respectively. We discovered commonalities and differences of gene expression profiles and molecular pathways between different cardiomyopathies. The gene discovery and molecular signature analysis in this study could offer insights into disease mechanisms and also identify markers useful for diagnostic, prognostic, and therapeutic purposes.

Gene expression changes in chicken NLRC5 signal pathway associated with in vitro avian leukosis virus subgroup J infection.

Nucleotide-binding oligomerization domain-like receptors (NLRs) play a key role in the innate immune response as pattern-recognition receptors. However, the role of NLRC5, which is a member of the NLR family, in NF-κB activation and MHC-I expression remains debatable. Infection with the J group avian leukosis virus (ALV-J) can result in immunosuppression and a subsequent increase in susceptibility to secondary infection. This results in huge economic losses to the poultry industry worldwide. Using quantitative real-time polymerase chain reaction (qRT-PCR), we investigated the mRNA expression levels of NLRC5 signal pathway-related genes in secondary chicken embryo fibroblasts 7 days after infection with ALV-J. The results indicated that, compared with the control groups, the expression levels of TLR7, MHC-I, and IL-18 increased significantly in the infected groups at 7 days post-infection (d.p.i.). The expression levels of NLRC5 and IL-6 were conspicuously downregulated at 7 d.p.i., but the expression levels of NF-κB, STAT1, and STAT3 were not significantly altered. These results suggest that NLRC5 and some genes involved in the NLRC5 pathway play a key role in antiviral immunity, typically the response to ALV-J infection. Moreover, MHC-I expression levels vary between different cell types.

Characterization of novel nitrate reductase-deficient mutants for transgenic Dunaliella salina systems.

The aim of the present study was to isolate and characterize novel nitrate reductase (NR)-deficient mutants, which may be useful for the transgenic manipulation of Dunaliella salina. Three NR-deficient mutants of D. salina, J-1, J-2, and J-3, were successfully isolated by screening for chlorate resistance after chemical mutagenesis with ethylnitrosourea. NR activity was not detected in the mutants and the expression of NR mRNA was significantly decreased. Growth analysis of D. salina strains grown in media containing different nitrogen sources revealed that these mutants were capable of utilizing nitrite and urea, but not nitrate as a nitrogen source, indicating that these mutants are indeed NR-deficient. Mutation analysis of NR cDNA sequences revealed that there were 11 point mutations shared by the J-1, J-2, and J-3 mutants. Furthermore, the results of the functional complementation experiment showed that NR activity of transformant T-1 derived from J-1 was recovered to 48.1 % of that of the wild-type D. salina. The findings of the present study indicate that nitrate may be used as a selective agent rather than antibiotics or herbicides for the isolated NR-deficient mutants in future transgenic D. salina systems.

Neurobehavioural abnormalities induced by repeated exposure of neonatal rats to sevoflurane can be aggravated by social isolation and enrichment deprivation initiated after exposure to the anaesthetic.

BACKGROUND: We tested the hypothesis that developmental effects of repeated neonatal exposure to sevoflurane in rats are exacerbated by stressful experiences received later in life. METHODS: Sprague-Dawley male rats received sequential exposures to 3% sevoflurane for two h on postnatal days (P) six, seven, and eight. After weaning at P21, rats were housed either in pairs in an enriched environment (EE) or singly in an enrichment-deprived environment (an adverse environment, AE). The hippocampal concentrations of brain-derived neurotrophic factor (BDNF), and synaptic markers were assessed at P8 and P53. The dentate gyrus neural progenitor proliferation was evaluated at P11 and P53 after administration of bromodeoyuridine (BrdU) at P8 to P10 and at P22 to P27, respectively. Neurobehavioural evaluations were performed at P49 to P53. RESULTS: Repeated sevoflurane exposure acutely reduced concentrations of BDNF, synaptic markers and neural progenitor proliferation. The sevoflurane group housed in the AE conditions (sevoflurane+AE) had decreased concentrations of BDNF and synaptic markers, and survival of new granule cells and impaired cognitive function compared with the control+AE, control+EE, and sevoflurane+EE groups. The neurobehavioural parameters in the sevoflurane+EE and control+EE groups were similar. CONCLUSIONS: Neurocognitive abnormalities induced by repeated neonatal exposure to sevoflurane can be aggravated by stressful conditions such as social isolation and enrichment deprivation.

Repeated ketamine administration redeems the time lag for citalopram's antidepressant-like effects.

Current available antidepressants exhibit low remission rate with a long response lag time. Growing evidence has demonstrated acute sub-anesthetic dose of ketamine exerts rapid, robust, and lasting antidepressant effects. However, a long term use of ketamine tends to elicit its adverse reactions. The present study aimed to investigate the antidepressant-like effects of intermittent and consecutive administrations of ketamine on chronic unpredictable mild stress (CUMS) rats, and to determine whether ketamine can redeem the time lag for treatment response of classic antidepressants. The behavioral responses were assessed by the sucrose preference test, forced swimming test, and open field test. In the first stage of experiments, all the four treatment regimens of ketamine (10mg/kg ip, once daily for 3 or 7 consecutive days, or once every 7 or 3 days, in a total 21 days) showed robust antidepressant-like effects, with no significant influence on locomotor activity and stereotype behavior in the CUMS rats. The intermittent administration regimens produced longer antidepressant-like effects than the consecutive administration regimens and the administration every 7 days presented similar antidepressant-like effects with less administration times compared with the administration every 3 days. In the second stage of experiments, the combination of ketamine (10 mg/kg ip, once every 7 days) and citalopram (20 mg/kg po, once daily) for 21 days caused more rapid and sustained antidepressant-like effects than citalopram administered alone. In summary, repeated sub-anesthestic doses of ketamine can redeem the time lag for the antidepressant-like effects of citalopram, suggesting the combination of ketamine and classic antidepressants is a promising regimen for depression with quick onset time and stable and lasting effects.

A novel amber mutation in a beta zero-thalassaemia gene (beta 37TGG-->TAG), with direct detection by mapping the restriction fragments in amplified genomic DNA.

A novel amber mutation, a G to A substitution at the second position of codon 37 in the beta-globin gene that changes the tryptophan coding triplet (TGG) to a termination codon (TAG), was found in a Chinese beta-thalassaemia carrier. The mutant gene creates an additional Dde I recognition site and eliminates the Ava II site, so this point mutation can be directly identified by restriction enzyme analysis.