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【Objective】 To investigate the efficacy of in vitro local high-frequency hyperthermia combined with drugs in the treatment of type Ⅲb chronic prostatitis (CP). 【Methods】 A total of 148 patients with type Ⅲb CP treated in our hospital during Jun.2020 and Jun.2022 were randomly divided into control group (n=74) and combination group (n=74). The control group received only drug treatment, while the combination group received extracorporeal local high-frequency hyperthermia treatment. The National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI), maximum urinary flow rate (MFR), average urinary flow rate (AFR) and International Index of Erectile Function-5 (IIEF-5) score were compared between the two groups. 【Results】 There were no significant differences between the two groups in terms of NIH-CPSI, MFR, AFR and IIEF-5 score before treatment(P>0.05). After treatment, these indexes increased in both groups (P<0.05), and the improvements in the combination group were more significant (P<0.05). 【Conclusion】 Medication combined with in vitro local high-frequency hyperthermia is effective in improving the clinical symptoms of type Ⅲb CP, which is worth clinical promotion.
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【Objective】 To compare the efficacy of transurethral columnar balloon dilation of the prostate (TUCBDP) and transurethral resection of prostate (TURP) in the treatment of small volume prostatic hyperplasia. 【Methods】 A total of 96 patients with small volume prostatic hyperplasia diagnosed in our hospital during Jan.2019 and Jan.2021 were enrolled and divided into the observation group and control group,with 48 patients in either group. The observation group received TUCBDP while the control group TURP. The International Prostate Symptom score (IPSS),Quality of Life Score (QOL), international index of erectile function-erectile function (IIEF-EF),maximum urinary flow rate (Qmax),postvoid residual urine (PVR) and maximum detrusor pressure (MDP) of the two groups were compared before surgery and 24 months after surgery. The surgery-related complications and occurrence of new or aggravated sexual dysfunction were observed. 【Results】 Both groups successfully completed the treatment. The operation time and indwelling catheterization time were shorter in the observation group than in the control group (P<0.05). The scores of QOL,IPSS and IIEF-EF,the levels of Qmax,PVR and MDP of both groups 24 months after surgery were significantly improved compared with those before surgery (P<0.05). The IPSS score of the observation group was lower than that of the control group 24 months after surgery (P<0.05),while the IIEF-EF score and Qmax of the observation group were higher than those of the control group (P<0.05). The incidences of surgery-related complications and new or aggravated sexual dysfunction were significantly lower in the observation group than in the control group (P<0.05). 【Conclusion】 TUCBDP is significantly effective in the treatment of small volume prostatic hyperplasia,showing greater advantages than TURP in improving postoperative IPSS,IIEF-EF score and Qmax,with higher safety.
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Prenatal corticosteroid for fetal lung maturation has been widely used in daily practice in obstetrics. However, previous research showed that prenatal corticosteroids may increase the risk of neonatal hypoglycemia and birth weight loss. And it remains controversial regarding the dosage and repeated administration. Based on the guideline "Antenatal corticosteroids to reduce neonatal morbidity and mortality" published by Royal College of Obstetricians and Gynaecologists in February 2022, here we are going to discuss several issues on the usage of prenatal corticosteroids.
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Objective: To evaluate the effectiveness of mucosal flap combined with silicone keel for preventing and treating anterior commissure adhesion in canines and clinical cases. Methods: A prospective experiment was performed from November 2019 to June 2021. Twenty five canines were randomly divided into 5 groups(A, B, C, D, E). Group A, B, C, D received anterior commissure injury by CO2 laser, then separately treated with free mucosal flap-keel complex,intralaryngeal mucosal flap-keel complex, silicone keels and without treatment, group E didn't injure the vocal cord after intubation. The keel was removed after 2 weeks, the larynx was harvested after 4 weeks. The effectiveness of anterior commissure adhesion prevention was evaluated by manifestation under laryngoscope, standard vocal cord length and standard glottic area. A retrospective analysis was performed on sixteen patients with anterior commissure lesion, who underwent mucosal flap-keel technique in Huashan Hospital of Fudan University from January 2019 to January 2021 (10 cases with free mucosal flap-keel complex and 6 cases with intralaryngeal mucosal flap-keel complex). All the patients underwent evaluation of laryngeal function included manifestation under laryngoscope each month and voice analysis before and 3 month after surgery. SPSS 20.0 software was used for statistical analysis. Results: No surgery accident or complication happened in canines and patients. The standard vocal cord length and standard glottic area after 4 weeks in group B were significantly higher than those in group A, C, D (Hstandard vocal cord length=31.688, Hstandard glottic area=16.444, P<0.05). The standard vocal cord length and standard glottic area after 4 weeks in group A were also significantly higher than those in group C, D(Hstandard vocal cord length=20.936, Hstandard glottic area=11.786, P<0.05). The standard vocal cord length and standard glottic area after 4 weeks in group A, B, E were not significantly different to that before surgery(tA left standard vocal cord length=2.636, tA right standard vocal cord length=2.582, tB left standard vocal cord length=2.707, tB right standard vocal cord length=2.673, tE left standard vocal cord length=0.370, tE right standard vocal cord length=0.821, tA standard glottic area=2.731, tB standard glottic area=2.753, tE standard glottic area=-0.529, P>0.05). The standard vocal cord length and standard glottic area after 4 weeks in group C, D were significantly lower than those before surgery(tC left standard vocal cord length=16.137, tC right standard vocal cord length=13.984, tD left standard vocal cord length=11.903, tD right standard vocal cord length=14.587, tC standard glottic area=10.280, tD standard glottic area=22.974, P<0.05). During 6-18 months of follow-up in clinical patients, no one developed a glottic web. Three months after surgery, Jitter, Shimmer, noise to harmonic ratio(NHR), the maximum phonation time(MPT)in all patients were significantly different from preoperative(tintralaryngeal mucosal flap jitter=24.885, tintralaryngeal mucosal flap shimmer=22.643, tintralaryngeal mucosal flap NHR=6.202, tintralaryngeal mucosal flap MPT=-9.661, tfree mucosal flap jitter=25.459, tfree mucosal flap shimmer=18.683, tfree mucosal flap NHR=5.705, tfree mucosal flap MPT=-20.840, P<0.05). Conclusion: Mucosal flap combined with silicone keel is an effective technique for preventing and treating anterior commissure adhesion. The effect of pedicled intralaryngea lmucosal flap is better.
Subject(s)
Animals , Dogs , Humans , Free Tissue Flaps , Glottis , Laryngeal Neoplasms/surgery , Prospective Studies , Retrospective Studies , Vocal Cords/surgeryABSTRACT
OBJECTIVE@#Taking the classic bilateral puncture vertebroplasty as a reference, to evaluate the clinical efficacy of vertebroplasty of the curved-angle puncture device, analyze the radiation exposure of patients and surgeons during the operation and summarize the protective measures.@*METHODS@#The clinical data of 49 patients with osteoporotic vertebral compression fractures admitted from March 2018 to September 2019 were retrospectively analyzed. According to the different surgical puncture methods, 49 patients were divided into vertebroplasty group (using classic bilateral puncture) and curved vertebroplasty group (using curved angle puncture). Among them, there were 26 cases in vertebroplasty group, including 7 males and 19 females, aged (73.25±6.36) years, 2 cases in thoracic segment, 21 cases in thoracolumbar segment, and 3 cases in lumbar segment. In curved vertebroplasty group, there were 23 cases, including 6 males and 17 females, aged (73.09±6.52) years, 3 cases in thoracic segment, 19 cases in thoracolumbar segment, and 1 case in lumbar segment. The operation time and the amount of injected bone cement in the two groups were recorded. Visual analogue scale (VAS) and modified Oswestry Disability Index (ODI) were respectively used to assess the pain degree and lumbar function, the postoperative bone cement leakage or other complications were observed. The radiation doses of the two groups of patients and surgeons were compared.@*RESULTS@#All 49 patients were followed up for 10-22 (14.55±3.83) months. Eleven cases in vertebroplasty group and 9 cases in curved vertebroplasty group occurred bone cement leakage after surgery, and there was no statistically significant difference between two groups. VAS scores of vertebroplasty group were 6.23±0.68 before operation and 1.69±0.47 at 1 day after operation, respectively, modified ODI were (72.59±3.25)% and (33.59±2.85)%. The preoperative and postoperative VAS scores of curved vertebroplasty group were 6.46±0.56 and 1.57±0.49, respectively, modified ODI were (73.21±3.18)% and (33.17±2.37)%. The postoperative pain degree and lumbar function of the two groups were significantly improved, but the difference between the groups was not statistically significant. The operation time of curved vertebroplasty group and vertebroplasty group were (17.27±9.58) min and(23.19±8.56) min, and the amount of injected bone cement were (4.91±1.49) ml and (6.58±1.42) ml. Obviously, curved vertebroplasty group has more advantages in operation time and the amount of injected bone cement. In curved vertebroplasty group, the radiation dose of the operator was (0.53±0.05) mSv and the patient was (10.64±1.65) mSv;in vertebroplasty group, the operator was (0.59±0.08) mSv andthe patient was (13.52±1.81) mSv. The radiation dose of patients in curved vertebroplasty group was significantly lower than that of the vertebroplasty group, but there was no statistically significant difference in the operator between two groups.@*CONCLUSION@#Both puncture methods can achieve satisfactory clinical results, but curved angle puncture can optimize the distribution of bone cement and reduce the radiation dose of patients.
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Female , Humans , Male , Fractures, Compression/surgery , Radiation Exposure , Retrospective Studies , Spinal Fractures/surgery , Treatment Outcome , VertebroplastyABSTRACT
OBJECTIVE@#To investigate the application of tranexamic acid in the treatment of intertrochanteric fracture.@*METHODS@#From January 2017 to October 2019, 100 patients with intertrochanteric fracture were randomly divided into observation group (48 cases) and control group(52 cases). All patients received the same surgical treatment. The control group was given tranexamic acid 20 minutes before operation, and 15 mg/kg diluted in 250 ml sodium chloride injection, intravenous drip;the observation group was given tranexamic acid 0.5 g dissolved in 20 ml normal saline injected into femoral bone marrow cavity for local treatment on the basis of the control group. The blood loss, operation time and postoperative hospital stay were compared between two groups. Hematocrit, hemoglobin, D-dimer and fibrinogen levels were analyzed before and after operation, and the incidence of thrombotic complications was observed.@*RESULTS@#The total blood loss, dominant blood loss, hidden blood loss and postoperative drainage volume of the observation group were significantly lower than those of the control group (@*CONCLUSION@#Tranexamic acid combined with systemic and local application has important clinical significance in reducing perioperative blood lossand blood cell loss in patients with intertrochanteric fracture, and has good safety.
Subject(s)
Humans , Antifibrinolytic Agents/therapeutic use , Blood Loss, Surgical , Femur , Hip Fractures/surgery , Postoperative Hemorrhage , Tranexamic Acid , Treatment OutcomeABSTRACT
Objective To explore the application of support vector regression (SVR) model combined with meteorological and air pollutants index in the prediction of the cases of bacillary dysentery in Lanzhou City, so as to provide scientific reference for the prevention and control of bacillary dysentery.Methods Time series data of the reported cases of bacillary dysentery from December 2013 to August 2016, combined with the meteorological and air pollutants data, were used as training set to fit support vector regression model. The data from September 2016 to December 2017 was used as validation set to verify the model and compare the effect in fit and prediction with different models. Results A total of 7 192 bacillary dysentery cases were reported in Lanzhou City from 2013 to 2017. The correlation coefficient of meteorological and pollution factors with the cases of bacillary dysentery was more than 0.4, except air pressure. The parameters of the fit model were selected based on the integrated data, acquiring the three parameters with the smallest test error were C=5, γ=0.02 and ε=0.000 1, respectively. The validation set was used to test the different models, which showed that the integrated data model had the best predictive accuracy and robustness . The root mean squared error (RMSE) was 0.164 7 and the mean absolute percentage error (MAPE) was 16.405%. Conclusion SVR model combined with meteorological and air pollutants index is effective in the prediction of bacterial dysentery.
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Lysinuric protein intolerance (LPI) is an autosomal recessive disorder caused by SLC7A7 gene mutation and often involves severe lesions in multiple systems. Lung involvement is frequently seen in children with LPI and such children tend to have a poor prognosis. This article summarizes the clinical manifestations and gene mutation characteristics of three children diagnosed with LPI by SLC7A7 gene analysis. All three children had the manifestations of aversion to protein-rich food after weaning, delayed development, anemia, hepatosplenomegaly, and osteoporosis, as well as an increase in orotic acid in urine. In addition, interstitial pneumonia and diffuse pulmonary interstitial lesions were observed in two children. SLC7A7 gene detection showed three pathogenic mutations in these children, namely c.1387delG(p.V463CfsX56), c.1215G>A(p.W405X) and homozygous c.625+1G>A. After a definite diagnosis was made, all three children were given a low-protein diet and oral administration of citrulline [100 mg/(kg.d)], iron protein succinylate [4 mg/(kg.d)], calcium and zinc gluconates oral solution (10 mL/day) and vitamin D (400 IU/day). In addition, patient 3 was given prednisone acetate (5 mg/day). The children had varying degrees of improvement in symptoms and signs. It is hard to distinguish LPI from urea cycle disorder due to the features of amino acid and organic acid metabolism in LPI, and SLC7A7 gene analysis is the basis for a definite diagnosis of LPI.
Subject(s)
Child , Humans , Amino Acid Metabolism, Inborn Errors , Genetics , Citrulline , Fusion Regulatory Protein 1, Light Chains , Genetics , Lysine , MutationABSTRACT
@#【Objective】To test the hypothesis that inhibiting sodium absorption via the epithelial sodium channel（ENaC） will increase ocular hydration and cure the rabbit′ s dry eye induced by scopolamine. 【Methods】In the experiment，24 New Zealand rabbits weighing about 2.0- 2.5 kg were divided into 3 groups：tested group（8 rabbits）， control group （8 rabbits），and treatment group （8 rabbits）. For the rabbits in the tested group，they were given subcutaneous injection of scopolamine 4 times a day for 12 consecutive days to induce the dry eye. For the rabbits in the control group，they accepted subcutaneous injection of saline solution. For the rabbits in the treatment group ，they were firstly given the same treatment as the rabbits in the tested group. Then，the local regions in their right eyes received the application of 100 mmol/L amiloride （a sodium channel inhibitor）. Meanwhile，the local regions in their left eyes received the application of equivalent saline solution. We detected ENAC α and ENAC γ subunits of epithelial sodium channel in conjunctival epithelium by fluorescence quantitative PCR and detected the opening of epithelial sodium channel by short- circuit current technology. Finally，we compared the ENaC α and ENaC γ gene expression，corneal fluorescein sodium staining and tear secretion among the 3 groups. Fluorescence quantitative PCR was used to detect ENaC α and ENaC γ subunits of epithelial sodium channel in conjunctival epithelium and short-circuit current was used to detect the opening of epithelial sodium channel. The ENaC α and ENaC γ gene expression，corneal fluorescein sodium staining and tear secretion （immerged length） were compared among 3 groups.【Results】 The results showed that the quantity of tear secretion was（17.00 ± 0.37）mm for the control group，（4.42 ± 1.34）mm for the tested group（P<0.001 vs Control，n=8） and（14.25 ± 0.54）mm for the treatment group（P>0.05 vs Control，n=8）. The results of short-circuit current detection showed that the sodium current was（5.72 ± 0.35）μA /cm2 in normal model and（12.24 ± 0.54）μA /cm2 in dry eye model（P<0.001）. After Amiloride treatment，the sodium current decreased to（4.00 ± 0.61）μA/cm2 （P>0.05） and there was no statistical difference compared with the normal group. According to the results of fluorescence quantitative PCR，the expression of ENaC α and ENaC γ subunits and IL- β in in dry eye model were up-regulated（P<0.01） compared with that in normal group. After topical amiloride application，there were no statistical differences in inflammatory cytokines IL- 1 β，ENaC α，and ENaC γ between the normal group and treatment group（P>0.05，n=8）. After treatment，tear secretion increased （P<0.001），and ocular surface staining improved significantly.【Conclusion】 Topical application of amiloride increase the quantity of preocular tears owing to inhibition of conjunctival sodium channels and could provide an effective new therapy for chronic dry eye.
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OBJECTIVE@#To investigate the effects of endothelial progenitor cells (EPCs) under shear stress on the biological function such as proliferation, adhesion, migration, apoptosis and expression of α-smooth muscle actin (α-SMA), collagen-I and collagen-Ⅲ of hepatic stellate cells (HSCs).@*METHODS@#HSCs and EPCs were inoculated into the upper and lower layers of the co-culture chamber respectively and co-incubated for 24 hours. Then, 12 dyne/cm shear stress was applied to EPCs cells for another 24 hours. After that, proliferation, adhesion, migration and apoptosis of HSCs were detected by cell counting kit-8 (CCK-8) kit, cell adherent assay, Boyden cell migration assay and flow cytometry respectively. Fluorescence quantitative PCR and Western blot were used to detect the mRNA and protein expression of alpha -SMA, collagen I and collagen-Ⅲ in HSCs.@*RESULTS@#Under shear stress, EPCs ecological niche could obviously inhibit the proliferation, adhesion and migration of HSCs, promote the apoptosis of HSCs, and down-regulate the mRNA and protein expression of collagen-I, collagen-Ⅲ in HSC cells.@*CONCLUSIONS@#Under shear stress, EPCs ecological niche could inhibit the fibrosis development of HSCs to a certain extent.
Subject(s)
Actins , Apoptosis , Cell Proliferation , Cells, Cultured , Collagen Type I , Endothelial Progenitor Cells , Hepatic Stellate CellsABSTRACT
BACKGROUND: Although more and more accurate and personalized detection and assistive tools are used in the field of orthopedics with the development of technology, the preoperative CT is still the first-line examination choice of many orthopedic surgeons. Therefore, to master the lumbar spine CT imaging performance is not only the basis of spine surgeons, but also the cornerstone of spinal nail surgery. OBJECTIVE: Through CT imaging technique, we measured the parameters of lumbar pedicle imaging, and understood its internal structure as the basis for the choice of screws, in order to improve the safety and accuracy of nail surgery. METHODS: Totally 36 cases without lumbar vertebrae-related disease undergoing lumbar vertebra and abdominal cavity CT scan were randomly selected, including 20 males and 16 females at the age of (43.3±12.3) years. PACS was used to reconstruct lumbar vertebrae, and the related imaging parameters were measured. RESULTS AND CONCLUSION: (1) There was no significant difference between the left and right sides of the pedicle for paired t-test (P >0.05). (2) Pedicle sagittal angle had downward trend, the level of change was small; L4increased relative to L3; L5level was minimum at the average of -1.3°. Pedicle transverse angle had gradually increased trend, and maximum at L5; the average was 25.4°. (3) The length of the pedicle channel increased first and then decreased in the lumbar segment; the longest average was 52.2 mm in the L2segment, the shortest at L5, with an average of 47.4 mm. (4) The pedicle height was slightly narrowed at L1-L3levels; the smallest was 15.4 mm (L3mean), and reached the maximum in the L5at 22.7 mm (L5mean). Cancellous bone height also showed the same trend, the minimum in the L3at 10.2 mm, the maximum in the L5at 17.9 mm. (5) Lumbar pedicle width showed an upward trend, L1minimum of 7.7 mm, L5maximum of 12.5 mm. Cancellous bone width also became a similar upward trend; the narrowest L1, averagely 4.9 mm; the maximum L5, averagely 8.5 mm. (6) Overall, the upper lumbar pedicle channel was slender, while the lower lumbar spine was slightly short. (7) The parameters of the pedicle can be used as the basis for selecting the appropriate screw. At the same time, lumbar pedicle morphology has a certain difference, and there is a certain law. Accurately grasping its structure and image characteristics of pedicle screw fixation is the basis of surgery. The preoperative imaging data collection and assessment, combined with the general situation of patients, and the development of personalized programming, will help in improving the safety and accuracy of screw placement.
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OBJECTIVE@#To explore the clinical phenotype, genetic variant, treatment and prognosis of a child with mosaic variegated aneuploidy syndrome (MVAS).@*METHODS@#Immunological marker screening, chromosomal karyotyping and whole exome sequencing were carried out.@*RESULTS@#The 1-year-11-month old girl has featured severe growth retardation, feeding difficulty, short stature, microcephaly, facial anomalies, scoliosis, visual impairment, hypotonia, chylothorax, and renal lesions. Karyotype analysis of peripheral blood lymphocytes has discovered variegated aneuploidy cells (6/11). DNA sequencing has identified compound heterozygous c.826delG (p.Asp276Metfs*21) and c.2441G>A (p.Arg814His) variants in the BUB1B gene, which were inherited from her father and mother, respectively.@*CONCLUSION@#The compound heterozygous variants of the BUB1B gene probably underlie the pathogenesis in this patient.
Subject(s)
Female , Humans , Infant , Aneuploidy , Chromosome Disorders , Diagnosis , Genetics , DNA Mutational Analysis , Genetic Testing , MosaicismABSTRACT
Objective To investigate the clinical efficacy of etoposide and levofloxacin in the treatment of chronic nonbacterial prostatitis(CNP)alone or in combination.Methods A total of 168 patients with CNP in our hospital from August 2014 to January 2016 were enrolled in this study.The patients were divided into A, B and C groups according to the random number table,56 cases in each group.Group A treatment alone, group B was treated with levofloxacin alone, group C was treated with Yongqing Tablet and levofloxacin, and the clinical efficacy and adverse hair loss were compared.Results After a course of treatment, there was no significant difference in the effective rate of treatment between group A and group B.The effective rate of treatment of group C was higher than that of group A and B, the difference was statistically significant (P<0.05).There was no significant differences in the incidence of adverse events between the three groups.Conclusion The clinical effect of Tongueqing and levofloxacin in the treatment of CNP is higher than that of single administration, and it will not increase the adverse reaction.It is worthy to be popularized and applied.
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Objective To investigate the effects of SMYD3 and MLL5 on histone methylation of Transcription factor forkhead box protein 3 (Foxp3) gene and its roles in the immunological pathogenesis of Kawasaki disease (KD). Methods Forty-two children with KD and 26 age-matched healthy children were consented to participate in this study. Co-Immunoprec-ipitation and real-time polymerase chain reaction (PCR) was performed to determine Foxp3-associated histone methylation levels of H3K4me3 and H3K27me3, and binding levels of SMYD3 and MLL5 with Foxp3 gene in CD4+T cells. The proportion of CD4+CD25high Foxp3+cells (Treg) and protein levels of Foxp3, cytotoxic T lymphocyte associated antigen-4 (CTLA4), TGF-βRⅡand pSmad3 were analyzed by flow cytometry. Quantitative real-time PCR was used to evaluate levels of Foxp3, interleukin (IL)-10, GITR, TGF-βRⅠand RARαmRNA in CD4+T cells. Plasma concentrations of TGF-βand retinol acid (RA) were measured by enzyme-linked Immunosorbent assay. Independent-samples t-test was used as the statistical method in this study. Results ① The proportion of Treg, expression levels of Foxp3 and molecules associated with suppressive function of Treg cells(IL-10, GITR and CTLA4), and histone methylation levels of H3K4me3 associating with promoter, conserved non-coding DNA sequence (CNS) 1 and CNS2 of Foxp3 gene decreased remarkably during acute KD [Promoter:(5.4±1.8)%vs (9.1±2.2)%;CNS1:(2.6±0.9)% vs (3.8±1.1)%; CNS2: (2.4±0.8)% vs (4.2±1.0)%; t=5.50, 6.02, 9.56, 7.92, 7.97, 4.76, 7.73, 5.01, 8.66; P0.05). ② Binding levels of SMYD3 and MLL5 with Foxp3 gene in CD4+T cells were down-regulated significantly during acute KD (t=6.63, 6.15; P<0.05), and restored to some extent after IVIG treatment (t=5.36, 4.56; P<0.05). Positive correlations between binding levels of SMYD3 and MLL5 and expression level of Foxp3 mRNA were detected in patients with acute KD (r=0.62、0.45, P<0.05). Furthermore, Binding levels of SMYD3 and MLL5 with Foxp3 gene in KD-CAL+group were lower than those in KD-CAL- group (t=4.11, 4.31; P<0.05). ③ Compared with healthy controls, plasma concentration of TGF-β and RA, and expressions of TGF-βRⅡ, TGF-βRⅠ, pSmad3 and RARα were down-regulated during acute KD (t=11.54, 12.81, 7.43, 16.10, 8.25, 12.06; P<0.05), and elevated remarkably after IVIG treatment (t=8.40, 6.24, 5.94, 11.78, 6.27, 8.30; P<0.05). Simultaneously, all the items aforementioned in KD-CAL+ group were found to be lower than those in KD-CAL-group (t=3.58, 3.30, 3.82, 5.27, 4.71, 3.78; P<0.05). Conclusion Hypomethylation of H3K4me3 associated with Foxp3 gene caused by insufficient binding levels of SMYD3/MLL5 may be involved with immune dysfunction in Kawasaki disease.
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Objective To investigate the effects of p300/CBP on histone acetylation of Foxp3 gene and its roles in the immunological pathogenesis of Kawasaki disease (KD).Methods Forty-six children with KD and twenty-eight age-matched health children were consented to participate in this study.Co-immunoprecipitation and real-time PCR were performed to detect Foxp3-associated acetylation levels of histone H4 and binding abilities of p300, CBP, pSmad3 (phosphorylated mothers against decapentaplegic homolog 3) and NF-AT (nuclear factor of activated T cells) with Foxp3 gene in CD4+ T cells.The percentages of CD4+CD25high Foxp3+ cells (Treg) and the expression of Foxp3, CTLA4 (cytotoxic T-lymphocyte-associated protein 4), p300, CBP, TGF-βRⅡ (transforming growth factor β receptor Ⅱ) and pLAT1 at protein level were analyzed by flow cytometry.Quantitative real-time PCR was used to measure the expression of Foxp3, IL-10, TGF-β, TGF-βRⅠ, Egr-1 (early growth response protein 1), RARα (retinoic acid receptor α) and PLCγ1 (phospholipase C-γ1) in Treg cells at mRNA level.Plasma concentrations of TGF-β and retinol acid (RA) were measured by enzyme-linked immunosorbent assay.Results (1) The percentages of Treg cells, levels of Foxp3 and molecules associated with suppressive function of Treg cells (TGF-β, IL-10 and CTLA4), acetylation levels of histone H4 associated with promoter, conserved non-coding DNA sequence 1 (CNS1) and CNS2 of Foxp3 gene decreased remarkably during acute KD (P0.05).(2) The levels of p300 and CBP in Treg cells and their binding abilities with Foxp3 gene were down-regulated significantly during acute KD (P0.05).Conclusion Hypoacetylation of histone H4 associated with Foxp3 gene caused by insufficient expression of p300/CBP and their impaired binding abilities might be involved with immune dysfunction in KD.IVIG therapy regulates the expression of p300/CBP and their binding abilities with Foxp3 gene through up-regulating TGF-β signal.
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Objective To investigate the clinical and biochemical metabolic features of 12 patients with systemic primary carnitine deficiency(CDSP) and to identify the SLC22A5 gene mutation types of the disease. Method The clinical and biochemical data were collected by retrospective analysis. DNA direct sequencing and multiplex ligation dependent probe amplification(MLPA)were applied for SLC22A5 gene analysis. Result Among 12 patients with CDSP, 3 cases had evident infection factors, 6 cases with convulsions, 5 cases manifested liver hypertrophy, 8 cases with hyperammonemia, and 9 cases showed myocardial damage. All CDSP patients were detected biallelic pathogenic mutation in SLC22A5 gene by direct sequencing. The gene types include IVS2+1G>T, c.3G>T(p.Met1Ile), c.760C>T(p.Arg254X), c.1400C>G(p.Ser467Cys), c.844dupc(p.Arg282fs), c.338G>A(p.Cys113Tyr), c.51C>G(p.Phe17Leu), c.659A>T(p.Glu220Val), and c.1365dupC(p.Thr456fs). c.659A>T(p.Glu220Val) and c.1365dupC(p.Thr456fs)are novel mutations. One female patient was maternal CDSP, her child had abnormal newborn screening. The allele frequency of c.760C>T(p.Arg254X) and c.1400C>G(p.Ser467Cys) were 37.5%(9/24)and 29.2%(7/24)respectively. The MLPA test results of all patients were negative. Conclusion The clinical manifestations are complex and various in patients with CDSP. Point and small InDel(insertions/deletions)mutation constitute the major alteration in SLC22A5 gene. c.1400C>G(p.Ser467Cys) might be another prevalence mutation type in Chinese CDSP patient.
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Objective@#To investigate the impacts of ash2 (absent, small, or homeotic)-like (Ash2L) and Jumonji domain-containing protein 3 (Jmjd3) on histone methylation of interferon-gamma(IFN-γ) gene and association with vascular damage of Kawasaki disease (KD) in acute phase.@*Methods@#This study was performed among 36 children with KD in acute phase (KD group) and 28 age-matched health children (control group), who were treated or underwent physical examination in our hospital between February 2015 and June 2016. Patients were further divided into KD groups with or without coronary artery lesions (KD-CAL+ , 16 cases; KD-CAL-, 20 cases). All KD patients were treated with intravenous immunoglobulin. The proportion of type 1 helper T(Th1) cells and protein levels of IFN-γ, T-box expressed in T cells(T-bet), phosphorylated signal transducer and activator of transcription 1(pSTAT1) and phosphorylated signal transducer and activator of transcription 4(pSTAT4) were analyzed by flow cytometry.Chromatin immunoprecipitation was performed to determine histone methylation (histone H3 tri-methyl K4(H3K4me3), histone H3 tri-methyl K27(H3K27me3)) and binding levels of Ash2L, Jmjd3 and Ezh2 associated with IFN-γ in CD4+ T cells. Quantitative real-time PCR was used to determine mRNA levels of IFN-γ, interferon γ receptor 1(IFN-γR1), interferon γ receptor 2(IFN-γR2), interleukin 12 receptor subunit beta 1(IL-12Rβ1), interleukin 12 receptor subunit beta 2(IL-12Rβ2), interleukin 18 receptor subunit beta α(IL-18Rα), interleukin 18 receptor subunit beta β(IL-18Rβ), tumor necrosis factor receptor 1(TNFR1), toll-like receptor 4(TLR4), receptor interacting serine/threonine kinase 1(RIP-1) and myeloid differentiation primary response gene 88(MyD88) in CD4+ T cells. Plasma concentrations of IFN-γ, interleukin 12(IL-12), interleukin 18(IL-18) and tumor necrosis factor α(TNF-α) were measured by enzyme-linked Immunosorbent assay.@*Results@#(1)The proportion of Th1 and its protein level of IFN-γ were significantly higher in KD group than those in control group and higher in KD-CAL+ group than in KD-CAL- group (all P<0.05), and lower after treatment than before treatment (all P<0.05). (2)Compared with control group, mRNA level of IFN-γ and IFN-γ-associating H3K4me3 was increased, while level of IFN-γ associating H3K27me3 in CD4+ T cells was reduced in KD group (all P<0.05), which resulted in a higher rate of H3K4me3/H3K27me3 (P<0.05) in KD group, which was positively correlated with IFN-γ mRNA in KD group(r=0.55, P<0.05). Similar results were found between KD-CAL+ group and KD-CAL- group (all P<0.05). Level of IFN-γ associating H3K27me3 was increased, and mRNA level of IFN-γ and IFN-γ associating H3K4me3 was decreased after treatment than before treatment (all P<0.05). (3)Expression of T-bet protein and binding levels of Ash2L and Jmjd3 with IFN-γ gene were significantly higher in KD group than those in control group(all P<0.05), higher in KD-CAL+ group than those in KD-CAL- group (all P<0.05). These parameters were significantly lower after treatment than before treatment (all P<0.05). Binding level of Ezh2 with IFN-γ gene was similar among various groups (all P>0.05). (4)In comparison with control or after treatment, surface receptors(IFN-γR1/2, IL-12Rβ1/2, IL-18Rα/β, TNFR1 and TLR4) and its downstream molecules(pSTAT1, pSTAT4, RIP1 and MyD88) in CD4+ T cells, and plasma concentrations of inflammatory cytokines(IFN-γ, IL-12, IL-18 and TNF-α) were found to be higher in KD group(all P<0.05). These parameters were also higher in KD-CAL+ group than in KD-CAL- group (all P<0.05).@*Conclusion@#Aberrant histone methylation of IFN-γ associating H3K4me3 and H3K27me3 caused by over-binding of Ash2L and Jmjd3 might be involved in immune dysfunction and vascular damage in KD in the acute phase.
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Objective The aim of present study was to detect methylation rate of CpG unit of brain derived neurotrophic factor (BDNF) promoter and to study the epigenetic mechanism of autism spectrum disorders (ASD).Methods Total of 12 ASD patients and 12 healthy controls were recruited.The methylation rate of CpG unit in BDNF promoter Ⅰ and Ⅳ were detected using Sequenom MassArray method.The methylation model,correlationship,evolutionary relationship of CpG units in BDNF promoter Ⅰ and Ⅳ were detected and compared between ASD patients and healthy controls.Results The methylation rate was identified in 17 and 8 CpG units in BDNF promoter][and BDNF promoter Ⅳ.A close correlation distance was detected in BDNF promoter Ⅰ CpG units 4,7,10,35,and BDNF promoter Ⅳ CpG units 11.12,14.BDNF promoter][CpG units 4,7,10,35,and BDNF promoter Ⅳ CpG units 11.12,14 could be clustered.ASD patients had a significant lower methylation rate in BDNF promoter Ⅰ CpG unit 5.6 and Ⅳ CpG units 3 and 15 compare with healthy controls (P＜0.05).Conclusions The DNA methylation rate in BDNF pronoter Ⅰ CpG unit 5.6 and Ⅳ CpG units 3 and 15 may be used as potential biomarkers of ASD.
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<p><b>OBJECTIVE</b>To analyze the clinical and molecular features of a child with carnitine palmitoyltransferase 1A (CPT1A) deficiency.</p><p><b>METHODS</b>Clinical data of the child was collected. Blood acylcarnitine was determined with tandem mass spectrometry. DNA was extracted from the child and his parents. All exons and flanking regions of the CPT1A gene were analyzed by PCR and Sanger sequencing.</p><p><b>RESULTS</b>Analysis showed that the patient carried compound heterozygous mutations c.1787T>C and c.2201T>C of the CPT1A gene, which derived his father and mother, respectively. Both mutations were verified as novel through the retrieval of dbSNP, HGMD and 1000 genome databases. Bioinformatic analysis suggested that the mutations can affect protein function.</p><p><b>CONCLUSION</b>Acyl carnitine analysis has been the main method for the diagnosis of CPT1A deficiency. The c.1787T>C and c.2201T>C mutations of the CPT1A gene probably underlie the disease in this patient. Gene testing can provide important clues for genetic counseling and prenatal diagnosis.</p>
Subject(s)
Female , Humans , Infant , Male , Pregnancy , Base Sequence , Carnitine O-Palmitoyltransferase , Genetics , Exons , Hypoglycemia , Genetics , Lipid Metabolism, Inborn Errors , Genetics , Molecular Sequence Data , Point MutationABSTRACT
This study demonstrates efficacy of a novel polyamidoamine dendrimers (PAMAM dendrimers) with pentaerythritol derivatives as the core (G5 PD dendrimer) in deliver of the cytosine deaminase (CD) gene and EGFP gene fusion plasmid into different tumor cell lines to induce apoptosis. The physical and chemical properties of G5 PD dendrimers in terms of DNA complexation, particulate properties and transfection efficiencies were investigated and compared with commercial gene vectors PEI 25 kDa. The optimum ratio of G5 PD dendrimer complexed with plasmid DNA was 0.2:1, and the particle size of the complex was (100±5) nm. Compared with the commercial gene carriers PEI, G5 PD dendrimer exhibited a higher transfection efficiency at the weight ratio of 1:1 in three different cell lines, given the fact that PEI are different from PAMAM dendrimers in terms of molecular structure. Furthermore, the cytotoxicity assays of the cell lines transfected with G5 PD dendrimer/pCD-EGFP-N1 followed by exposure to various concentrations of 5-fluorocytosine (5-FC) also showed that the transfected cell lines could generate a very low amount of 5-FC to 5-fluorouracil (5-FU) in a short period of time, which indicating the high expression level of CD gene in the cell line. These results indicate that the CD/5FC system of G5 PD dendrimer has an excellent efficacy in gene delivery.