OBJECTIVE: To explore the genetic etiology of a Chinese pedigree with recurrent Joubert syndrome with negative results by whole-exome sequencing in the prior proband. METHODS: Chinese pedigree which opted elective abortion at the Women and Children's Hospital Affiliated to Chongqing Medical University in December 2024 was selected as the study subject. Whole-genome sequencing was carried out on fetal tissue after termination of pregnancy. Candidate variants were validated by Sanger sequencing and interpreted, while non-coding variant was analyzed using in silico prediction tools. RNA sequencing and cDNA sequencing were conducted on fetal brain tissue. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.2024YL045-02). RESULTS: Both the fetus and the affected child were found to harbor compound heterozygous variants of the CEP290 gene, namely c.7341dup (p.Leu2448fs*8) (pathogenic, maternally inherited) and c.1523-408G>A (likely pathogenic, paternally inherited). Both in silico analysis and fetal brain RNA sequencing confirmed aberrant RNA splicing caused by the intronic variant. CONCLUSION This case has highlighted the value of combining whole-genome sequencing with RNA functional validation. Above results not only enriched the spectrum of CEP290 gene mutations but also underscored its diagnostic value in resolving complex prenatal cases, providing critical clues for the prenatal diagnosis and recurrence risk assessment in genetic counseling.
Female ; Humans ; Pregnancy ; Abnormalities, Multiple/genetics* ; Antigens, Neoplasm/genetics* ; Cell Cycle Proteins/genetics* ; Cerebellum/abnormalities* ; Cytoskeletal Proteins/genetics* ; Eye Abnormalities/genetics* ; Introns/genetics* ; Kidney Diseases, Cystic/diagnosis* ; Pedigree ; Retina/abnormalities* ; Sequence Analysis, RNA/methods* ; Whole Genome Sequencing/methods* ; Child

OBJECTIVE To explore the clinical characteristics of Tropheryma whipplei(TW)infection and observe the application of pathogenic metagenomic next-generation sequencing(mNGS)in diagnosis of TW infection.METHODS The clinical data were collected from 1 patient who was diagnosed by mNGS in the Affiliated Hospital of Jining Medical University on Apr.9,2022.The data including the results of laboratory tests and treatment out-comes were summarized,and a literature review was conducted.RESULTS A 50-year-old woman presented to the hospital with chest tightness and chest pain lasting for 3 days,accompanied by dyspnea,palpitations,and expec-toration.The chest plain CT scan and magnetic resonance imaging(MRI)scan suggested a high probability of pul-monary infection.Normal flora were isolated by culture of bronchoalveolar lavage fluid(BALF);TW and human βherpes virus type 7 were detected in BALF by mNGS,with the sequence numbers 327 000 and 9,respectively.The pulmonary symptoms of the patient were improved after joint treatment of the infection with etimicin,levo-floxacin and minocycline.The patient repeatedly sought for medical treatment due to the pain of shoulder joint and limitation of motion.CONCLUSIONS TW is one of major pathogens leading to the infections of systemic multiple systems,and it is necessary to attach great importance to the diagnosis and treatment.The traditional laboratory test method can not achieve ideal diagnosis effect and is more likely to make a missed diagnosis.mNGS is more ac-curate and more efficient than the traditional detection method in diagnosis of TW-induced diseases.Early use of mNGS can make a rapid identification of pathogens and facilitate the reasonable clinical use of antibiotics.It is of great significance for control of the disease progression,improvement of prognosis and prevention of recurrence.

Objective:To explore the genetic etiology of a Chinese pedigree with recurrent Joubert syndrome with negative results by whole-exome sequencing in the prior proband.Methods:A Chinese pedigree which opted elective abortion at the Women and Children′s Hospital Affiliated to Chongqing Medical University in December 2024 was selected as the study subject. Whole-genome sequencing was carried out on fetal tissue after termination of pregnancy. Candidate variants were validated by Sanger sequencing and interpreted, while non-coding variant was analyzed using in silico prediction tools. RNA sequencing and cDNA sequencing were conducted on fetal brain tissue. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.2024YL045-02). Results:Both the fetus and the affected child were found to harbor compound heterozygous variants of the CEP290 gene, namely c. 7341dup (p.Leu2448fs*8) (pathogenic, maternally inherited) and c. 1523-408G>A (likely pathogenic, paternally inherited). Both in silico analysis and fetal brain RNA sequencing confirmed aberrant RNA splicing caused by the intronic variant. Conclusion:This case has highlighted the value of combining whole-genome sequencing with RNA functional validation. Above results not only enriched the spectrum of CEP290 gene mutations but also underscored its diagnostic value in resolving complex prenatal cases, providing critical clues for the prenatal diagnosis and recurrence risk assessment in genetic counseling.

OBJECTIVE To explore the clinical characteristics of Tropheryma whipplei(TW)infection and observe the application of pathogenic metagenomic next-generation sequencing(mNGS)in diagnosis of TW infection.METHODS The clinical data were collected from 1 patient who was diagnosed by mNGS in the Affiliated Hospital of Jining Medical University on Apr.9,2022.The data including the results of laboratory tests and treatment out-comes were summarized,and a literature review was conducted.RESULTS A 50-year-old woman presented to the hospital with chest tightness and chest pain lasting for 3 days,accompanied by dyspnea,palpitations,and expec-toration.The chest plain CT scan and magnetic resonance imaging(MRI)scan suggested a high probability of pul-monary infection.Normal flora were isolated by culture of bronchoalveolar lavage fluid(BALF);TW and human βherpes virus type 7 were detected in BALF by mNGS,with the sequence numbers 327 000 and 9,respectively.The pulmonary symptoms of the patient were improved after joint treatment of the infection with etimicin,levo-floxacin and minocycline.The patient repeatedly sought for medical treatment due to the pain of shoulder joint and limitation of motion.CONCLUSIONS TW is one of major pathogens leading to the infections of systemic multiple systems,and it is necessary to attach great importance to the diagnosis and treatment.The traditional laboratory test method can not achieve ideal diagnosis effect and is more likely to make a missed diagnosis.mNGS is more ac-curate and more efficient than the traditional detection method in diagnosis of TW-induced diseases.Early use of mNGS can make a rapid identification of pathogens and facilitate the reasonable clinical use of antibiotics.It is of great significance for control of the disease progression,improvement of prognosis and prevention of recurrence.

Objective:To explore the genetic etiology of a Chinese pedigree with recurrent Joubert syndrome with negative results by whole-exome sequencing in the prior proband.Methods:A Chinese pedigree which opted elective abortion at the Women and Children′s Hospital Affiliated to Chongqing Medical University in December 2024 was selected as the study subject. Whole-genome sequencing was carried out on fetal tissue after termination of pregnancy. Candidate variants were validated by Sanger sequencing and interpreted, while non-coding variant was analyzed using in silico prediction tools. RNA sequencing and cDNA sequencing were conducted on fetal brain tissue. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.2024YL045-02). Results:Both the fetus and the affected child were found to harbor compound heterozygous variants of the CEP290 gene, namely c. 7341dup (p.Leu2448fs*8) (pathogenic, maternally inherited) and c. 1523-408G>A (likely pathogenic, paternally inherited). Both in silico analysis and fetal brain RNA sequencing confirmed aberrant RNA splicing caused by the intronic variant. Conclusion:This case has highlighted the value of combining whole-genome sequencing with RNA functional validation. Above results not only enriched the spectrum of CEP290 gene mutations but also underscored its diagnostic value in resolving complex prenatal cases, providing critical clues for the prenatal diagnosis and recurrence risk assessment in genetic counseling.

Objective:To investigate the characteristics of neointimal hyperplasia (NIH) in patients with in-stent restenosis (ISR) over 5 years post-drug-eluting stent (DES) implantation based on optical coherence tomography (OCT).Methods:In this cross-sectional study, patients with DES-ISR who underwent OCT examination at PLA General Hospital between March 2010 and March 2022 were retrospectively included. All patients were divided into≤5 years DES-ISR group and>5 years DES-ISR group according to the time interval after DES implantation. Quantitative and qualitative analyses were conducted on OCT images to compare the clinical data and lesion characteristics of two patient groups. Furthermore, the independent clinical predictive factors of in-stent neoatherosclerosis (ISNA) were analyzed by multivariable logistic regression.Results:A total of 230 DES-ISR patients with 249 lesions were included, with an age of (63.1±10.4) years and 188 males (81.7%). The median interval after DES implantation was 6 (2, 9) years. There were 117 patients (122 ISR lesions) in the≤5 years DES-ISR group, and 113 patients (127 ISR lesions) in the>5 years DES-ISR group. Compared with≤5 years DES-ISR,>5 years DES-ISR showed more heterogeneous patterns (65.4% (83/127) vs. 48.4% (59/122), P=0.007), diffuse patterns (46.5% (59/127) vs. 31.2% (38/122), P=0.013), macrophage accumulations (44.1% (56/127) vs. 31.2% (38/122), P=0.035) in NIH and higher prevalence of ISNA (83.5% (106/127) vs. 72.1% (88/122), P=0.031). According to multivariable logistic regression, the independent predictive factor for ISNA was female ( OR=0.44, 95% CI 0.21-0.90, P=0.026). Female ( OR=0.48, 95% CI 0.23-0.99, P=0.046) and low-density lipoprotein cholesterol level ( OR=1.62, 95% CI 1.01-2.59, P=0.046) were independent predictive factors, respectively, for lipid ISNA. Calcified ISNA was independently associated with time interval of post-DES implantation ( OR=1.18, 95% CI 1.07-1.29, P=0.001). Conclusion:DES-ISR patients with a time interval of>5 years after stent implantation have a higher prevalence of ISNA and more complex lesions. Gender, the level of low-density lipoprotein cholesterol, and the time interval post-DES implantation are independently correlated with ISNA, lipid ISNA, and calcified ISNA.

Epilepsies are a group of chronic neurological disorders characterized by spontaneous recurrent seizures caused by abnormal synchronous firing of neurons and transient brain dysfunction. The underlying mechanisms are complex and not yet fully understood. Endoplasmic reticulum (ER) stress, as a condition of excessive accumulation of unfolded and/or misfolded proteins in the ER lumen, has been considered as a pathophysiological mechanism of epilepsy in recent years. ER stress can enhance the protein processing capacity of the ER to restore protein homeostasis through unfolded protein response, which may inhibit protein translation and promote misfolded protein degradation through the ubiquitin-proteasome system. However, persistent ER stress can also cause neuronal apoptosis and loss, which may aggravate the brain damage and epilepsy. This review has summarized the role of ER stress in the pathogenesis of genetic epilepsy.
Humans ; Endoplasmic Reticulum Stress/genetics* ; Unfolded Protein Response ; Endoplasmic Reticulum/pathology* ; Apoptosis ; Epilepsy/genetics*

Objective:To summarize and analyze the clinical data of Chinese patients with colony-stimulating factor 1 receptor (CSF1R)-related leukoencephalopathy, and clarify the phenotypic and genetic characteristics of Chinese patients.Methods:Medical history of patients with CSF1R-related leukoencephalopathy diagnosed from April 1, 2018 to January 31, 2021 in the department of neurology of 22 hospitals in China was collected, and scores of Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment Scale (MoCA), magnetic resonance severity scale were evaluated. Group comparison was performed between male and female patients.Results:A total of 62 patients were included, and the male-female ratio was 1∶1.95. The age of onset was (40.35±8.42) years. Cognitive impairment (82.3%, 51/62) and motor symptoms (77.4%,48/62) were the most common symptoms. The MMSE and MoCA scores were 18.79±7.16 and 13.96±7.23, respectively, and the scores of two scales in male patients (22.06±5.31 and 18.08±5.60) were significantly higher than those in females (15.53±7.41 , t=2.954, P=0.006; 10.15±6.26, t=3.328 , P=0.003). The most common radiographic feature was bilateral asymmetric white matter changes (100.0%), and the magnetic resonance imaging severity scale score was 27.42±11.40, while the white matter lesion score of females (22.94±8.39) was significantly higher than that of males (17.62±8.74 , t=-2.221, P<0.05). A total of 36 CSF1R gene mutations were found in this study, among which c.2381T>C/p.I794T was the hotspot mutation that carried by 17.9% (10/56) of the probands. Conclusions:The core phenotypic characteristics of CSF1R-related leukoencephalopathy in China are progressive motor and cognitive impairment, with bilateral asymmetrical white matter changes. In addition, there exist gender differences clinically, with severer cognitive impairment and imaging changes in female patients. Thirty-six CSF1R gene mutations were found in this study, and c.2381T>C/p. I794T was the hotspot mutation.

Objective:To evaluate the value of mammography and MRI combined with clinical features in predicting upgrade to malignancy in high-risk breast lesions.Methods:Data from 230 patients who were diagnosed with high-risk breast lesions and underwent both mammography and MRI examinations before biopsy were analyzed retrospectively from Jan 2017 to Mar 2018 in Fudan University Shanghai Cancer Hospital. The imaging features of both mammography and MRI were analyzed, and the association between mammography, MRI and clinical features were evaluated using pathology as the gold standard. Independent t test and χ 2 test were used to compare the difference of clinical and imaging features between upgrade and non-upgrade groups, using receiver operating characteristic (ROC) curve to test the diagnostic value between mammography and MRI. Binary logistic regression was used to evaluate the correlation between upgrade and clinical, imaging findings. Results:Two hundred and thirty patients had 230 lesions, and 47 cases had atypia upgrade to malignancy during second surgery (upgrade rate was 20.4%). There were statistically significant differences in age, maximum diameter of lesion, and menopausal status between the upgraded and non-upgraded groups ( P<0.05). There was no statistically significant difference in mammographic features between two groups ( P>0.05), while there was statistically significant difference in breast MRI features and background parenchymal enhancement ( P<0.05). For the diagnostic value in predicting upgrade of high-risk lesions, MRI was better than mammography (the areas under ROC curve were 0.913 and 0.606, Z=6.919, P<0.01). Single factor analysis showed that age, lesion size, menopausal status, MRI negative and background parenchymal enhancement on MRI were significantly different for upgrade to malignancy ( P<0.05). Multiple factors analysis showed age and background parenchymal enhancement on MRI were independent factors for predicting upgrade ( P<0.01). Conclusion:For the upgrade to malignancy in high-risk lesions, the diagnostic value of MRI is better than mammography. The elder age and moderate or marked background parenchymal enhancement on MRI may serve as useful predictors of upgrade.

Objective To investigate the mechanism of white matter damage (WMD) and the neuroprotective effect of Xenon on neonates with WMD.Methods Three-day-old SD rat pups (n =96) were randomly divided into the blank control group (n =24),the WMD control group (n =24),the Xenon intervention group A (n =24) and the Xenon intervention group B (n =24) by random number method according to their birth time.WMD rat models were successfully established by giving intraperitoneal injection of lipopolysaccharide(LPS) 0.05 mg/kg combined with carotid artery ligation and hypoxia for 1 hour in the WMD control group and the Xenon intervention groups.In the control group,only 9 g/L saline (0.05 mg/kg) was injected intraperitoneally,while carotid artery ligation and hypoxia were not administered.Rats in Xenon intervention group A and group B were given inhalation of 500 mL/L Xenon for 3 hours at 0 and 2 hours respectively after establishment of the models.Six rats in each group were randomly selected and decapitated at 0,24,48 and 72 hours after the intervention.The brain white matter on the right was analyzed by using HE staining and myelin basic protein(MBP) immunofluorescence staining,and real-time quantitative polymerase chain reaction was used to detect the expressions level of CLIC4 mRNA.Results (1) Brain tissue pathology:compared with the blank control group,the brain white matter on the right of the WMD control group and the Xenon intervention group A and group B had loose and disordered structure,nuclear pyknosis and cytoplasm loosening.However,the lesions in both Xenon intervention group A and group B were significantly less than those in the WMD control group,and there was no significant difference between the Xenon intervention group A and group B.(2) MBP measurement:the number of MBP-positive cells in the brain white matter on the right of WMD control group was significantly lower than that in the blank control group,while compared with WMD control group,they were significantly higher in Xenon intervention group A and group B.(3) CLIC4 mRNA expression level:compared with blank control group,the expressions levels of CLIC4 mRNA at most time point were higher both in the WMD control group and the Xenon intervention group A and group B (all P ＜ 0.05),except the time point 24 h in the Xenon intervention group A.The expressions of CLIC4 mRNA in group A and group B were significantly decreased compared with those in the WMD control group (all P ＜ 0.05).However,there were no significant differences between Xenon intervention group A and group B (P ＞ 0.05).Conclusions The expressions of CLIC4 mRNA in brain tissues on neonatal rats with WMD significantly increased,indicating that the mitochondrial pathway could be one of the pathological processes of WMD.Early Xenon intervention may reduce neonatal WMD by reducing the expression of CLIC4 mRNA,which plays a neuroprotective role.