Objective:To apply digital analysis to quantify hyperechogenicity of substantia nigra, and explore its clinical value for diagnosis of Parkinson′s disease (PD).Methods:The cross-sectional study included 652 PD patients (PD group) and 99 healthy controls (healthy control group) from November 2017 to October 2020 in Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology. All subjects underwent transcranial sonography. The diagnostic accuracy of substantia nigra hyperechogenicity using digital analysis was compared with that in a manual measurement in PD. Furthermore, the receiver operating characteristic (ROC) curve analysis was applied to explore its diagnosis value in PD.Results:There were 482 subjects including 400 in the PD group and 82 in the healthy control group, whose quantified results of substantia nigra hyperechogenicity could be used for analysis. The ROC analysis showed that the area under the curve of the quantified larger substantia nigra hyperechoic region detection for diagnosing PD was 0.858 (95% CI 0.805-0.910), the sensitivity was 87.8%, and the specificity was 73.2%, consistent with that of doctors (area under the curve: 0.884). Further more, among these PD patients, there was no correlation between larger substantia nigra hyperechogenicity and age, age of onset, course of disease, non-motor symptoms, and motor symptoms (all P>0.05). Conclusions:Digital analysis was used to quantify the changes in substantia nigra hyperechogenicity in this seudy. The results showed that diagnostic accuracy for PD based on digital analysis was consistent with that of experienced clinicians.

OBJECTIVE To investigate the intervention effect and potential mechanism of breviscapine on hepatic fibrosis （HF） in rats based on the transforming growth factor-β（1 TGF-β1）/Smad2/extracellular signal-regulated protein kinase 1（ERK1） and Kelch-like epichlorohydrin-associated protein 1（Keap1）/nuclear factor-erythroid 2-related factor 2（Nrf2）/heme oxygenase-1（HO-1） pathways. METHODS Totally 60 rats were randomly divided into normal control group， model group， breviscapine low-dose， medium-dose and high-dose groups （5.4， 10.8， 21.6 mg/kg）， and colchicine group （positive control， 0.45 mg/kg）， with 10 rats in each group， half male and half female. Except for the normal control group， HF model of the other groups was induced by carbon tetrachloride. Subsequently， each drug group was given corresponding medicine by gavage once a day for 28 days. The liver appearance of rats in each group was observed and their liver coefficients were calculated. The levels of alanineaminotransferase （ALT） and aspartate aminotransferase （AST）in serum， those of ALT， AST， superoxide dismutase （SOD），malondialdehyde （MDA） and glutathione peroxidase （GSH- Px） in liver tissue were detected. The liver tissue inflammatory and fibrotic changes were observed. The protein and mRNA expressions of TGF-β1， Smad2， ERK1， Nrf2， Keap1 and HO-in liver tissue were detected. RESULTS Compared with the normal control group， the model group showed large areas of white nodular lesions in the liver， obvious inflammatory cell infiltration and collagen fiber deposition. The body weight， the levels of SOD and GSH-Px in liver tissue， the protein and mRNA expressions of Nrf2 and HO-1 were significantly lowered in the model group （P＜0.05）； the liver coefficient， the percentage of Masson staining positive area， ALT and AST levels of serum and liver tissue， MDA level of liver tissue， the protein and mRNA expressions of TGF-β1， Smad2， ERK1 and Keap1 were significantly increased （P＜0.05）. Compared with the model group， the liver lesions of rats in each drug group were improved， and the above quantitative indexes were generally reversed （P＜0.05）. CONCLUSIONS Breviscapine has a good intervention effect on HF rats， which may be related to inhibiting TGF-β1/Smad2/ERK1 pathway for anti-fibrosis and regulating Keap1/Nrf2/HO-1 pathway to inhibit oxidative stress.

Objective:To investigate the form of programmed cell death (PCD) induced by severe fever with thrombocytopenia syndrome virus (SFTSV) infection in Vero cells and further explore the existence of pyroptosis, so as to provide new ideas for studying the pathogenic mechanism of SFTSV.Methods:Vero cells were infected with SFTSV at different multiplicity of infection (MOI), cytopathic effect (CPE) was observed daily, cell viability was detected by CCK-8 method, and cell membrane damage was detected by LDH release test to determine the optimal amount of virus infection and cell death time. Vero cells were pretreated with different PCD inhibitors and infected with SFTSV. CCK-8 kit was used to detect the cell viability and determine the death form of PCD caused by SFTSV. The expression of pyroptosis related proteins was detected by Western blotting to further explore the existence of pyroptosis.Results:At 48 h and 72 h after SFTSV infected Vero cells with MOI=10, the optimal infection amount and time of subsequent experiments were observed. At this time, the CPE of cells was obvious, the cell viability decreased to 51% and 41% of the control group ( P<0.001, P<0.001), and the LDH release amount reached 24% and 37% of the maximum enzyme activity release wells, were 3.8, 3.4 times of LDH release in the control group ( P<0.001, P<0.001). The inhibition of SFTSV-induced cell death by different PCD inhibitors showed that pan-caspase inhibitor and receptor-interacting serine/threonine-protein kinase 3 (RIP3) inhibitor had inhibitory effects at 48 h and 72 h. The cell viability was 2.1 and 1.6 times of the viral control group at 48 h, 2.3 and 1.7 times of the viral control group at 72 h, and the effect of pan-caspase inhibitor was significantly higher than that of the other inhibitor groups. Caspase-1 and caspase-3 inhibitors only had inhibitory effect at 48 h, and the cell viability was 1.5 and 1.3 times of the viral control group. After SFTSV infection of Vero cells, the expressions of caspase-1 and IL-1β increased gradually with the prolongation of time, and reached 3.4 and 9.5 times of the control group at 48 h, respectively ( P<0.001, P<0.001). Conclusions:SFTSV infection of Vero cells can lead to various forms of PCD, including apoptosis, pyroptosis and programmed necrosis, and pyroptosis related protein activation can be detected in the process of PCD, which further explored the existence of pyroptosis.

Objective:To evaluate the efficacy and safety of a China-made disposable peroral cholangioscope in animal cholangioscopy.Methods:Six healthy Bama minipigs underwent the domestic disposable peroral cholangioscopy after anesthesia. The operation performance and image quality of the products were evaluated, and intraoperative and postoperative complications were recorded.Results:All 6 animals received the domestic disposable peroral cholangioscopy. The cholangioscope showed good operating performance and could smoothly enter the biliary tract through the duodenoscope. The water injection, suction and instrument channels were unobstructed. The cholangioscopic image was clear, the color resolution was good, and there was no image distortion. The lumen and the mucosal surface could be accurately assessed. During the examination, there was no operative injuries such as bleeding, perforation, or adverse events such as respiratory depression or cardiac arrest. All pigs survived the operation with no adverse reaction.Conclusion:The China-made disposable peroral cholangioscope is safe, with good operability and high-quality image.

Objective:To investigate the clinical, genetic, and clonality related aspects of individuals with Richter transformation (RT) .Methods:From January 2019 to December 2021, 18 RT patients with diagnoses at the First Affiliated Hospital of Nanjing Medical University (Pukou CLL center) were retrospectively examined. The immunoglobin heavy variable (IGHV) gene usage and IGHV-D-J rearrangement pattern of diagnosed CLL/SLL and transformed diffuse large B-cell lymphoma (DLBCL) were compared to determine the clonality relatedness. To investigate the risk factors of RT, Clinical and laboratory data from patients with newly diagnosed CLL/SLL and transformed DLBCL were gathered.Results:The median age of RT was 56.5 (41-75) years old. 17 patients transformed to DLBCL and 1 transformed to Hodgkin lymphoma (HL) . Of 17 individuals who had DLBCL transformation, 15 had CLL/SLL-related clonality and 2 had unrelated clonality. Next-generation sequencing (NGS) analysis of 11 paired initially diagnosed treatment-naive CLL/SLL and RT DLBCL found that EGR2、TP53 and NOTCH1 were among the most frequently mutated genes both in treatment-naive CLL/SLL and in RT DLBCL. In several cases, specific mutations were gained or lost throughout RT, indicating clonal evolution. Among 10 patients before exposure to BTK inhibitors before RT, four patients acquired BTK mutation. The aforementioned mutations should be considered high-risk variables for transformation; in addition, TP53 and EGR2 mutations could be linked to a poor prognosis following RT in patients receiving a cocktail of new medicines.Conclusion:Most RT DLBCL patients in our center are clonality related (15/17, 88.2% ) and we recommend all qualified centers to evaluate clonality relatedness of RT DLBCL patients. There was some variability in the mutational landscape between DLBCL that had undergone a transformation and initially diagnosed, treatment-naive CLL/SLL. The underlying molecular mechanism of RT needs more research.

OBJECTIVE: To analyze the clinical and genetic features of three patient diagnosed with Kleefstra syndrome. METHODS: Whole exome sequencing (WES) was carried out for the probands and their parents. Suspected variants were validated by Sanger sequencing. Copy number variations (CNV) were detected by CNV-seq and validated by real-time PCR. RESULTS: Proband 1 was found to carry a de novo heterogeneous variant (c.823+1G>T) of the EHMT1 gene, which may affect its expression. Based on the guidelines of the American College of Medical Genetics and Genomics, the variant was predicted to be pathogenic (PVS1+PS2+PM2). Proband 2 was found to carry a de novo missense variant c.439C>G (p.L147V) of the EHMT1 gene, which was predicted to be likely pathogenic (PS2+PM1+PM2+PP3). Proband 3 was found to carry a heterozygous 520 kb deletion at 9q34.3 by CNV-seq. The deletion has encompassed the whole of the EHMT1 gene. Real-time PCR has detected no CNV of this region in her parents. CONCLUSION Variants of the EHMT1 gene probably underlay the disease in these patients. Genetic testing has provided a basis for their clinical diagnosis.
Chromosome Deletion ; Chromosomes, Human, Pair 9 ; Craniofacial Abnormalities ; DNA Copy Number Variations ; Female ; Genetic Testing ; Heart Defects, Congenital ; Humans ; Intellectual Disability/genetics* ; Mutation

Objective:To investigate the status and influencing factors of skin cleaning outside wound (hereinafter referred to as skin) in adult trauma patients.Methods:A multicenter cross-sectional investigation was conducted. From September 1 to 30, 2020, a total of 952 adult trauma patients who met the inclusion criteria were admitted to wound care clinics or trauma surgery wards of 13 military or local Grade Ⅲ Level A hospitals, including the General Hospital of the Eastern Theater Command of People's Liberation Army and the Army Medical Center, etc. A self-designed questionnaire on cleaning status of skin in trauma patients was released through the "questionnaire star" website to investigate basic information such as gender, age, education level, living status, and self-care ability, trauma information such as cause of injury, wound duration, trauma site, trauma depth, wound pain, wound peculiar smell, and wound cleaning solution, and skin cleaning status after injury such as whether to clean or not, cleaning method, cleaning frequency, cleaning duration in each time, or reasons for not cleaning. The patients who cleaned skin regularly after injury were included in cleaning group, and the other patients were included in no cleaning group. The basic information, trauma information, and skin cleaning status after injury of patients in 2 groups were investigated. Data were statistically analyzed with chi-square test, and binary multivariate logistic regression analysis was performed on indicators with statistically significant differences between the two groups to screen the independent influencing factors of skin cleaning in trauma patients.Results:A total of 952 questionnaires were received, and the recovery rate was 100%. Three invalid questionnaires were eliminated, and 949 valid questionnaires were obtained, with an effective rate of 99.68%. In 949 patients, there were 461 (48.6%) males and 488 (51.4%) females, aged 18-100 (50±18) years. Most patients were less than 60 years old, lived with their families, and could take care of themselves completely. Nearly half of the patients were with junior high school or below education level. The main causes of injury were sharp cutting injury and falling injury, the wound duration was 2-365 days, most of the injured parts were limbs and trunk, the wound depth was mostly full-thickness injury, and most patients had wound-related pain and no peculiar smell and used 5 g/L iodophor to clean the wound. Totally 684 (72.1%) patients cleaned their skin after injury, mainly by scrubbing with warm water, the cleaning frequency was mainly once or twice a week, and the cleaning time was mainly 10 or 15 min for each time. Totally 265 (27.9%) patients didn't clean their skin after injury, and the main causes for not cleaning were following the doctor's advice, followed by worrying about wound infection and loss of self-care ability. There were significantly statistical differences in constituent ratios of education level, self-care ability, cause of injury, wound pain, and wound peculiar smell of patients in 2 groups ( χ2=12.365, 24.519, 22.820, 9.572, 92.342, P<0.01). Education level, self-care ability, cause of injury, wound pain, and wound peculiar smell were potential influencing factors of skin cleaning in patients. Binary multivariate logistic regression analysis showed that self-care ability, wound pain, and wound peculiar smell were independent influencing factors of skin cleaning in patients (odds ratio=1.51, 0.52, 3.72, 95% confidence interval=1.08-2.12, 0.42-0.89, 2.66-5.22, P<0.05 or P<0.01). Conclusions:Self-care ability, wound pain, and wound peculiar smell are independent influencing factors of skin cleaning in adult trauma patients.

Objective:To compare the performance and surgical outcomes of disposable digital flexible ureteroscope with a reusable fiberoptic flexible ureteroscope in treatment of upper urinary stones.Methods:A prospective, multicenter, single-blind and randomized controlled study was performed from April 2018 to December 2018. Eligible patients were randomized, in a ratio of 1∶1, to either experimental group or control group. The inclusion criteria for the study were: aged 18-60 years, solitary upper urinary stone or multiple stones with stone size less than 2 cm, absence of urinary tract infection (UTI) or UTI was controlled, normal liver and renal function. Exclusion criteria included: patients with congenital anomalies, calyceal diverticular stone, IP angle less than 30°, renal insufficiency. pregnancy or lactation, cardiopulmonary function abnormality and coagulation abnormalities. Patients in experimental group received ureteroscopy through a disposable digital flexible ureteroscope (PU3022A, PUSEN), while patients in control group received ureteroscopy through a reusable fiberoptic flexible ureteroscope (Flex-X2, STORZ). The primary endpoint was the high-quality rate of images during the operation. The secondary endpoints included success rate of fragmentation and postoperative complication rates.Results:93 patients were recruited, and 90 of them were finally complete the study (i.e., 45 patients in each group). The demographic and preoperative parameters were comparable between the two groups except the stone size in patients with solitary stone. The high-quality rate of images was significantly higher in experiment group than that in control group (100.0% vs. 15.6%, P<0.001). There were no significant differences in terms of success rate of fragmentation(68.9% vs. 71.1%, P=0.818), hemoglobin dropped (3.91 g/L vs. 3.62 g/L, P=0.880), serum creatinine changed, and postoperative complication rates(6.7% vs.6.7%, P=1.000). Conclusions:Disposable digital flexible ureteroscope achieves similar surgical outcomes to the reusable fiberoptic flexible ureteroscope with a better quality of endoscopic images.

OBJECTIVE: To delineate the clinical feature and genetic basis of four patients with congenital neutropenia. METHODS: All patients were subjected to whole exome sequencing (WES). Suspected variants were verified by Sanger sequencing. RESULTS: The patients (two boys and two girls), aged 7 to 15 months, suffered from neutropenia and recurrent infections. Bone marrow smears showed a significant decrease in the proportion of rod-shaped and lobulated granulocytes, which suggested impaired development and maturation of bone marrow neutrophils. WES has discovered heterozygous variants (c.496G>A, c.58C>G, c.391G>A and IVS1+5T>A) of the ELANE gene in the patients. Among these, c.58C>G and IVS1+5T>A were unreported previously. Follow up revealed patients 1 and 3 had periodic neutropenia, while patients 2 and 4 had severe congenital neutropenia. After attaining the definite diagnosis, the patients were treated symptomatically. CONCLUSION The main clinical feature of congenital neutropenia is refractory recurrent bacterial infections, for which mutations of the ELANE gene are a common cause. Two novel pathogenic ELANE variants have been discovered in this study.
Congenital Bone Marrow Failure Syndromes/genetics* ; Female ; Genetic Testing ; Humans ; Infant ; Leukocyte Elastase/genetics* ; Male ; Mutation ; Neutropenia/genetics*