Objective:To retrospectively analyze the clinical phenotype and pathogenic variants in patients with Progressive myoclonus epilepsy (PME).Methods:Clinical data and results of genetic testing for 11 patients diagnosed with PME at the Department of Neurology, the First Affiliated Hospital of Zhejiang University School of Medicine from June 2017 to December 2022 were collected and analyzed.Results:All of the patients, including 4 males and 7 females, had predominant action myoclonus. Three patients had myoclonus as the initial manifestation, whilst eight were diagnosed through genetic testing, including three cases with NEU1 gene variants, two with EPM2A gene variants (1 was novel), one with MT- TK gene variant, one with ATN1 gene variant, and one with CSTB gene variant. No pathogenic variant was identified in the remaining three cases. Among the eight patients with a genetic diagnosis, three were diagnosed with sialidosis, two with Lafora disease, one with Dentatorubral-pallidoluysian atrophy (DRPLA), one with Unverricht-Lundborg disease (ULD), and one with Myoclonic epilepsy with ragging red fibers (MERRF). Conclusion:Compared with pediatric patients, adult patients with PME represent a distinct subtype with slower progression and milder cognitive impairment.

OBJECTIVETo screen for CGG repeats in the FMR1 gene among patients with fragile X syndrome and carriers of pre-mutations.

METHODSPotential full and pre-mutations of the FMR1 gene were detected with a Tri-primer-florescence PCR-Sanger sequencing method. The results were validated with positive and negative controls.

RESULTSAll positive and negative controls were confirmed. A male patient was found to have > 200 CGG repeats (full mutation). For a pregnant women who was heterozygous for 35/115 CGG repeats, > 200 CGG repeats were also found with amniotic fluid sample from her fetus who was a male. The result was confirmed by following selective abortion with informed consent.

CONCLUSIONTri-primer-florescence PCR-Sanger sequencing is a simple, effective and reliable method for routine screening of patients/carriers with full/pre-mutations of the FMR1 gene in the population.

DNA Primers ; genetics ; Female ; Fluorescence ; Fragile X Mental Retardation Protein ; genetics ; Fragile X Syndrome ; genetics ; Humans ; Male ; Mutation ; genetics ; Polymerase Chain Reaction ; methods

Objective To construct pIRES2-ZsGreen1/FⅨ expression vector,using the pcDNA/ FⅨ plasmid containing FⅨ cDNA as template,and express in HEK-293 cells.Methods The total ORF of F Ⅸ gene was amplified from pcDNA/F Ⅸ plasmid,then the amplified fragment was clonded into the pIRES2-ZsGreen1 vector using the Infusion enzyme.The positive clones of eukaryotic expression vector of pIRES2-ZsGreen1/F Ⅸ were screened and expanded after transfection,then were constructed and confirmed by PCR and sequencing.Transient expression experiments were performed using HEK-293 cells transfected with the expression vectors and observed the expression of ZsGreenl protein by confocal laser microscope.The relative expression levels of F Ⅸ mRNA,protein and F Ⅸ activity (F Ⅸ ∶ C) were detected by real time PCR (RT-PCR),immunofluorescence microscopy,One-Stage method,respectively.Results The expression vector,pIRES2-ZsGreen1/F Ⅸ,was successfully constructed and expressed in HEK-293 cells.RT-PCR detected the expression of F Ⅸ mRNA in HEK-293 cells and the immunofluorescence microscopy showed F Ⅸ protein distributed in the surrounding of nucleus.F Ⅸ ∶ C of cell lysates and cell culture fluid transfected with the expression vectors were (92.03 ± 0.29)％ and (86.89 ± 8.78)％,respectively;while both F Ⅸ ∶ C of cell lysates and cell culture fluid transfected with or without the expression vectors were 0.Conclusion The experimental results showed the expression vector,pIRES2-ZsGreen1/F Ⅸ,was successfully constructed,which provided experiment basement for the follow study on the location,function and molecular pathology of hemophilia B.

Objective To investigate the effects of continuous positive airway pressure (CPAP) on the levels of hemorheology and C-reactive protein in patients with obstructive sleep hypopnea apnea syndrome (OSAHS). Methods 58 moderate to severe OSAHS subjects were selected as treatment group and 32 healthyadults were selected as control group. Hemorheology and C-reactive protein in all subjects were examined and compared. The levels of hemorheology and C-reactive protein were also examined and compared before and after 3 months CPAP treatment in treatment group. Results In treatment group, the levels of the whole blood viscosity and hs-CRP were significantly higher than those in control group (P < 0.01). After treatment of CPAP for three months, the levels of the whole blood viscosity and hs-CRP obviously decreased (P < 0.01). Conclusion Levels of hemorheology and hs-CRP are elevated in patients with OSAHS and CPAP therapy could effectively decrease serum levels of hemorheology and hs-CRP in patients with OSAHS.

Objective To explore the therapeutic effects of statin in patients with chronic obstructive pulmonary disease (COPD) in stable period. Methods 83 patients with COPD in stable period were recruited and randomly divided into two groups, treatment group (n = 41) and control group (n = 42).The control group received conventional routine therapy, and treatment group was given rosuvastatin calcium 20 mg once per day on the basis of routine treatment. The levels of sera C-reactive protein (CRP), pulmonary function (FEV1、FEVl%pred), and COPD assessment test (CAT) were observed at the time points of 0 week and 12th week, 24th week and 48th week were compared between two groups. Results At 12th week after the treatment, sera CRP levels, FEV1, FEV1%pred and CAT scores in either control group or treatment group showed some extent of improvements as compared to those at the time point of 0 week, and there was statistical difference of sera CRP levels and CAT scores between two groups (P < 0.05). There was no statistical difference of FEV1, FEV1%pred between two groups (P > 0.05) .24th week and 48th week after the treatment; sera CRP levels, FEV, FEV1%pred and CAT scores in two groups all significantly improved and were statistical different between two groups (P < 0.05). Conclusion Statin can alleviate inflammatory reaction, improve pulmonary function and life quality of patients with COPD.

Objective To summarize the experience in nursing children with pediatric advanced intraocular retinoblastoma treated with 23-gauge microinvasive vitrectomy. Methods Eleven infants (12 eyes) suffered from advanced intraocular (D or E) retinoblastoma and treated with 23-gauge microinvasive vitrectomy. Preoperative psychological nursing, postoperative complication nursing and follow-up nursing were performed. Results The retinal tumors was completely removed in all 12 eyes. After the follow-up of a half year, the conditions in 8 eyes were stable, and the tumors in 4 eyes recurred. Visual acuity in 4 eyes improved and that in 1 eye remained unchanged. Conclusions Microinvasive vitreous operation can effectively remove advanced intraocular retinoblastoma. Active efficient perioperative nursing plays an important role for the prevention of complications and promoting the recovery of affected eyes.

Objective To investigate the effect of shRNA-mediated down-regulation of the receptor for activated C kinase 1 (RACK1) gene on the chemotherapeutic sensitivities in human lung adenocarcinoma cell line A549.Methods The shRNA recombinant plasmid targeting to human RACK1 gene was designed and transferred into A549 cells by lipofectin technique.The protein level of RACK1 was measured by Western blot to confirm the function of shRNA plasmid.Drug sensitivities of A549 cells to cisplatin,gemcitabine,pemetrexed and paclitaxel were analyzed by MTT assay.The protein expression of LRP and MRP were detected by Western blot.Results After 24 hours transfection,the relative expression quantity of RACK1 protein in RACK1-shRNA group was 0.267± 0.470,which was significantly lower than that in vector-shRNA group (0.821±0.109) and control group (0.842±0.060) (F =54.438,P ＜ 0.05).The results of MTT showed that the growth of A549 cells in the RACK1-shRNA group was markedly inhibited.The sensitivities of A549 cells to cisplatin and paclitaxel were significantly enhanced compared with that in the vector-shRNA group and control group (P ＜ 0.05).The relative expression quantity of LRP and MRP protein in RACK1-shRNA group were 0.163±0.056 and 0.246±0.050,which were lower than that in vector-shRNA group and control group (F LRP =19.430,F MRP =61.548,both P ＜ 0.05).Conclusion Targeted gene silencing of RACK1 improves the sensitivity of A549 cells to the ascisplatin and paclitaxel medicines,which might be achieved through down-regulation of the expression of LRP and MRP.

Objective To investigate the effects of diversified health education in screening of preemy retinopathy. Methods One hundred and eighty preterm newborns were collected and randomly divided into control group and observation group on average. The diversified health education was adopted in observation group and conventional health education was adopted in control group. The disease cognition and satisfaction questionnaire investigated in the parents of preemys, and compared after 6 months′ intervention. Results After diversified health education, the rate of mastering knowledge of disease from parents in the observation group was higher than that in control group (95. 6% vs 80. 0% ) while the degree of satisfaction from parents in observation group was higher than that of the control group (90. 0% vs 71. 1% ) ( χ2 = 8. 751, 14. 754;P < 0. 05). Conclusions The development of diversified health education can effectively improve the disease cognition and the degree of satisfaction from parents. It can also enhance the consciousness of ROP screening of the parents and play an active role in ROP prevention.

BACKGROUNDThoracic endovascular aortic repair (TEVAR) is an emerging treatment modality, which has been rapidly embraced by clinicians treating thoracic aortic disease. However, the clinical manifestations of systemic inflammatory response after TEVAR as post-implantation syndrome (PIS) resemble the perioperative infection. This study aimed to evaluate changes and diagnostic value of procalcitonin (PCT) and other traditional inflammatory markers for infections after TEVAR.

METHODSWe conducted a prospective clinical study that enrolled 162 consecutive aortic dissection cases, who underwent TEVAR in our institution between July 2011 and November 2012. The PCT, C-response protein (CRP), erythrocyte sedimentation rate (ESR) and blood routine examination were monitored before the operation and on days 1, 2, 3 and 5 after the operation. The diagnosis of infection was confirmed by the infection control committee with reference to Hospital Acquired Infection Diagnostic Criteria Assessment, released by the Ministry of Health of the People's Republic of China.

RESULTSPost endovascular repair of thoracic aorta, PCT changes significantly at different time points (χ(2) = 13.225, P = 0.021), without significant difference between the PIS group and the control group (0.24 ± 0.04 vs.0.26 ± 0.10, P = 0.804). PCT values were significantly higher in the first day after TEVAR than the preoperative levels (0.18 ± 0.03 vs. 0.11 ± 0.02, P < 0.001). Compared with PIS patients, the level of PCT, CRP, White blood cell (WBC) and neutrophil (NEU) in the infection patients elevated significantly (relatively χ(2) = 6.062, P = 0.048; χ(2) = 6.081, P = 0.048; χ(2) = 11.030, P = 0.004; χ(2) = 14.632, P = 0.001). According to the ROC analysis, the PCT levels in the first day after TEVAR (AUC = 0.785, P = 0.012) had better predictive values of infection than WBC, NEU CRP and ESR (AUC = 0.720, P = 0.040; AUC = 0.715, P = 0.045; AUC = 0.663, P = 0.274; AUC = 0.502, P = 0.991). The best predictive index was the changes of PCT between preoperative and postoperative (PCT), which possess AUC as 0.803 (P = 0.014). And PCT = 0.055 could be considered as an infection diagnosis cutoff value with a sensitivity of 83.3% and specificity 69.0%.

CONCLUSIONSPCT provides better diagnostic value of infection compared with other inflammatory markers. The potential applications of PCT in differential diagnosis of PIS and infection after percutaneous TEVAR deserve further studies.

Adult ; Aged ; Blood Sedimentation ; C-Reactive Protein ; metabolism ; Calcitonin ; metabolism ; Calcitonin Gene-Related Peptide ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Protein Precursors ; metabolism ; Vascular Surgical Procedures