Genetic analysis and in vitro validation of a case of Alport syndrome due to a splicing variant of COL4A5 gene / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a patient with Alport syndrome (AS) and confirm the existence of a splicing variant.@*METHODS@#An AS patient diagnosed at the Affiliated Hospital of Inner Mongolia Medical University on January 8, 2021 for significant proteinuria and occult hematuria was selected as the study subject. Clinical data was collected. Peripheral blood samples were collected for the extraction of genomic DNA. Whole exome sequencing and Sanger sequencing were carried out to identify potential genetic variants. An in vitro experiment was also conducted to verify the abnormal mRNA splicing. Bioinformatic software was used to analyze the conservation of amino acids of the variant sites and simulate the 3D structure of the variant collagen IV protein. Immunofluorescence and immunohistochemistry were carried out on renal tissues from the patient to confirm the presence of AS kidney injury.@*RESULTS@#The patient, a 21-year-old male, had a 24-hour urine protein of 3.53 g/24 h, which fulfilled the diagnostic criteria for proteinuria. His blood uric acid has also increased to 491 μmol/L. DNA sequencing revealed that he has harbored a c.835-9T>A splice variant of the COL4A5 gene, which was not found in either of his parents. In vitro experiment confirmed that the variant has removed 57 bp from the exon 15 of the mRNA of the COL4A5 gene. The deletion may cause loss of amino acid residues from positions 279 to 297, which in turn may affect the stability of the secondary structure of the α5 chain encoded by the COL4A5 gene. The amino acids are conserved across various species. The result of homology modeling indicated that the trimerization of Col-IV with the mutated α5 chain could be achieved, however, the 3D structure was severely distorted. The AS kidney damage was confirmed through immunofluorescence assays. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.835-9T>A variant was classified as likely pathogenic (PVS1_Moderate+PS3_Moderate+PM2_Supporting+PS2+PP3+PP4).@*CONCLUSION@#The c.835-9T>A variant of the COL4A5 gene probably underlay the AS in this patient. In vitro experiment has confirmed the abnormal splicing caused by the variant. Histopathological examination of the kidney tissue has provided in vivo evidence for its pathogenicity. Above finding has expanded the mutational spectrum of the COL4A5 gene.

Evaluation of plasma exosomal miR-214-3p as a potential biomarker for diagnosis and prognosis of uveal melanoma / 中华实验眼科杂志

Objective:To investigate the differential expression of microRNA (miR)-214-3p in plasma exosomes in different types of uveal melanoma (UM) and evaluate whether miR-214-3p is a potential molecular biomarker for the diagnosis and prognosis of UM.Methods:Twenty-five UM in situ patients who received the enucleation of eyeball were enrolled at Beijing Tongren Hospital from December 2015 to October 2019, including 10 with epithelioid cell melanoma and 10 with spindle cell melanoma as well as 5 metastatic UM patients (1 with spindle cell-like melanoma and 4 with epithelioid cell-like melanoma) and 10 healthy subjects were enrolled during the same period.Blood sample was collected from all the subjects for the isolation of plasma exosomes.The morphology of exosomes was examined under the electron microscope.The exsomal marker proteins were identified by Western blot.The expression level of miR-214-3p in plasma exosomes was detected by real-time fluorescence quantitative PCR.The differential expression of miR-214-3p among different types of UM patients and healthy controls was compared.The diagnostic and classification performance of exosomal miR-214-3p was evaluated using receiver operating characteristic curve.Written informed consent was obtained from each subject prior to entering the study cohort.This study protocol was approved by an Ethics Committee of Capital Medical Univeristy (No.TRECKY2018-056).Results:The isolated exosomes were hemispherical in shape with a concavity on one side.The diameter of the exosomes was about 100 nm and the particle diameter of vesicles from samples was (82.0±2.7) nm.TSG101 protein was detectable and Calnexin protein was not found in the exosomes.The relative expression levels of plasma exosomal miR-214-3p in healthy control group, in situ UM group and metastatic UM group were 0.86(0.57, 1.49), 0.24(0.10, 0.67), and 0.43(0.23, 0.56), respectively.The miR-214-3p relative expression level in plasma exosomes of in situ UM patients and metastatic UM patients was significantly lower than that of healthy controls, and the differences were statistically significant ( Z=2.62, P<0.01; Z=2.08, P<0.05). The relative expression levels of exosomal miR-214-3p in spindle cell-like UM group and epithelioid cell-like UM group were 0.11(0.07, 0.64) and 0.46(0.14, 0.91), respectively, and no significant difference was found in the expression level of plasma exosomal miR-214-3p among different types of UM (all at P>0.05). The area under the curve of plasma exosomal miR-214-3p for UM diagnosis was 0.795. Conclusions:Plasma exosomal miR-214-3p level is significantly reduced in both in situ UM patients and metastatic UM patients.Plasma exosomal miR-214-3p is a new potential diagnostic biomarker for UM, but the exosomal miR-214-3p appears to not be able to distinguish the types of UM.

Treatment patterns of patients with multiple sclerosis in Guangzhou, China

@#Background & Objective: Disease-modifying treatments (DMTs) for multiple sclerosis (MS) are widely used in Western countries. In China, however, the current treatment patterns of MS patients are not well characterized. This is to explore the gap between the current treatments in Guangzhou, Southern China and those given in Western countries. Methods: We performed a survey of MS patients at department of neurology, a tertiary MS referral centre in Guangzhou, concerning treatments of MS in Southern China. The clinical data in patients were collected. The initial treatment, drug withdrawal or switching profile, and therapeutic effect of existing treatments in MS patients were analyzed. Results: The ratio of MS patients who receive DMTs in Guangzhou China is extremely low. Among the 178 patients studied, only 28.09% received initial treatment with DMTs. MS patients who receive initial treatment with first-line DMTs have higher drug withdrawal rates (32.6%) and drug switching rates (30.43%) than those of western populations. The main reasons for withdrawal of first-line DMTs were doctor’s advice (maintenance of remission) (40.00%), economic burden(20.00%), and no channels to buy drugs(13.33%). In MS patients initially treated with first-line DMTs who switched to other drugs, a gap between treatments was common (8/14;57.14%). There were 18 patients with highly active MS receiving treatment with rituximab. Annual relapse rate after treatment significantly decreased than that before treatment (0.74 vs. 1.50 , P < 0.001). Conclusions: DMTs for MS in Guangzhou, Southern China appear to lag behind those in Western countries. Much work is needed to improve drug accessibility and affordability of DMTs in China. Rituximab is an option for highly active MS in limited medical-resource countries.

One case of anti-NMDAR encephalitis combined with NMOSD during pregnancy and literature review / 中国医师杂志

Anti-N-methyl-D-aspartate receptor (N-methyl-D-aspartate receptor,NMDAR) encephalitis is the most common autoimmune encephalitis,mostly in young women of childbearing age,but how pregnancy influence the disease is still unclear.This article reports a case of gestational morbidity,and analyzes 10 pregnant women in different stages and their babies published in domestic and foreign,aiming to briefly explore the influences of different fetuses during pregnancy.

Risk factors for moderate and severe postoperative pain in patients undergoing percutaneous nephrolithotomy / 中华麻醉学杂志

Objective To determine the risk factors for moderate and severe postoperative pain in patients undergoing percutaneous nephrolithotomy.Methods Eighty-one patients of both sexes,aged 18-70 yr,with body mass index of 19-42 kg/m2,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients,undergoing elective percutaneous nephrolithotomy under general anesthesia,were selected.When moderate and severe postoperative pain developed (visual analog scale score ≥ 4),dezocine 5 mg was intravenously injected with the dosing interval greater than 4 h.According to the occurrence of moderate and severe pain within 24 h after operation,the patients were divided into moderate and severe pain group and non-moderate and severe pain group.The age,sex,American Society of Anesthesiologists physical status,body mass index,preoperative abnormality of creatinine,urinary leukocyte,urinary bacteria,surgical site,severity of hydronephrosis,the number of affected renal calyx,CT value of stones,length of puncture channel,operation time,the number of postoperative drainage tube and residual stones were collected.Multivariate logistic regression analysis was performed to stratify the risk factors for moderate and severe postoperative pain.Results A total of 79 patients were included in this study.Among the 79 patients,47 patients developed moderate or severe pain,and the incidence was 59％.The results of multivariate logistic regression analysis showed that the number of preoperative affected renal calyx＞3,CT value of stones ＞ 1 000 and postoperative residual stones were independent risk factors for moderate and severe pain after percutaneous nephrolithotomy (P＜0.05).Conclusion The number of preoperative affected renal calyx＞3,CT value of stones ＞ 1 000 and postoperative residual stones are independent risk factors for moderate and severe postoperative pain in the patients undergoing percutaneous nephrolithotomy.

Nuclear factor kappa B,Bcl-2 and Bax in limb function after acute spinal cord compression injury / 中华物理医学与康复杂志

Objective To investigate the relationship of nuclear factor kappa B(NF-κB),Bcl-2 and Bax with limb function after acute spinal cord injury in rats. Methods Forty-eight rats were divided at random into a control group and an experimental group with 24 rats in each.The spinal cords of the rats in the experimental group were injured at the T8,9,10 level through moderate compression.Four hours,8 h,and 1,3,7 and 14 days after the injury,4 rats were selected randomly from each group and graded with a BBB score.They were then sacrificed and their spinal cords were collected.Immunohistochemical measurements were used to observe the expression of NF-κB, Bcl-2 and Bax. Results NF-κB,Bcl-2 and Bax were observed in the injured spinal nerve cells of rats in the exper imental group but were absent in the control group.After injury,the expression of these factors increased at first and then decreased.BBB scores for limb function increased gradually.No correlation was found between the changes in NF-κB and Bcl-2,but the expression of NF-κB was positively correlated with that of Bax.There was negative correla tion between NF-kB levels and BBB scores,and between NF-kB levels and the ratio of Bcl-2 to Bax. Conclusion In rats,there is a close negative correlation between NF-kappa B levels,the ratio of Bcl-2/Bax and limb function after acute spinaI cord iujury.

Etiological study on hallux valgus of 1233 adults in Shijiazhuang / 中国矫形外科杂志

[Objective]To investigate the hallux valgus angle and pathogenesis of hallux valgus in general population.[Method]A total of 1 233 adult population in Shijiazhuang were selected for this study by random cluster sampling method,then a questionnaire survey was participated including gender,age,nation,culture degree,inhabitate time,symptome occurrence time,original symptome,aggravate factors,relieve factors,family medical history,living habit,life style,et al.Hallux valgus angle was measured by protracter.The angle exceed 15? was confirmed the hallux valgus.The data was analysised with logistic multiple regression analysis.[Result]The average hallux valgus angle was(8.31?4.93)? of male and(9.72?7.12)? of female.There were 98 hallux valgus patients in this study,and the morbidity rate was 7.95%.The morbidity rate was 1.29% of male and 11.00% of female.Ratio of male to female was 1∶8.53.The average age was 22 years old.The concentrated symptome occurrence time was 20 to 45 years old;the average age was 25 years old.78 hallux valgus patients had a family medical history(79.6%).The dangerous factors of hallux valgus were inheritance,gender,flatfoot.The effective factors in female related to inheritance,the age that start wearing high-heel shoes but not to the height of the heel and the full time who were high-heel shoes.The aggravate factors were as follow:wearing high-heel shoes,long time standing,long time walking.The relieving factors were as follow:wearing flat and low-hell shoes,a warm water bath for foot,massage.[Conclusion]Female has a high risk to have hallux valgus.The hallux valgus has a close relationship with inheritance,and the age who start wearing high-heel shoes influences the incidence in female.