ObjectiveTo observe the effects of human IL-10 gene transfection on the mRNA and protein expressions of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion injury in rats and to investigate its neuroprotective mechanism. MethodsRats were divided into four groups: normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group. The mRNA and protein expressions of IL-1β and TNF-α in the penumbra area were detected by fluorescence real-time quantitative PCR and ELISA respectively. ResultsIn normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group, the levels of protein expression of TNF-α in penumbra area were(0.66±0. 04) ,(1.16±0.26),(1. 155±0. 26)ng/g and(0. 84±0. 05)ng/g, and the levels of protein expression of IL-1βin penumbra area were(0.37±0.05), (1.25±0.39), (1.21±0.57) ng/g and(0.62+0.05)ng/g, respectively. Compared with normal control group, the levels of protein expression of TNF-α and 1L-1β were significantly higher in other three groups(all P＜0. 01), and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group(all P＜0. 01). In normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfectedgroup, the levels of mRNA expression of TNF-α in penumbra area were 1.00 ±0.53,9.42±1.83,9.69±1.96 and 3.53±1.09, and the levels of mRNA expression of IL-1β in penumbra area were 1.00 ±0.51,27. 81±4.84,23.96 ± 4.90 and 13.55± 4.45, respectively. Compared with normal control group, the levels of mRNA expression of TNF-α and IL-1β were significantly higher in other three groups(all P＜0. 01), and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group(all P＜0. 01). ConclusionsThe human IL-10 gene transfection may play an protective effect on cerebral ischemia through inhibiting mRNA and protein expression of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion in rats.

ObjectiveTo study the changes of serum immune state in maintenance hemodialysis (MHD) patients with high-flux hemodialysis. MethodsSixty MHD patients were divided into high-flux hemodialysis group (group T, 30 cases, receiving high-flux hemodialysis, 3 times per week and routine drug therapy) and conventional dialysis group (group C, 30 cases, taking conventional dialysis, 3 times per week and routine drug therapy) by random number table. They were treated for 12 weeks. Serum IgG,IgA, IgM,C3, C4, T lymphocyte subsets and albumin, prealbumin, hemoglobin were detected at the experiment onset and 12 weeks later, and compared with those of 20 normal medical examination adults (group N ), meanwhile the ratio of infection was calculated. ResultsThe levels of IgG,IgA,IgM,C3,C4,CD3+,CD4+,CD4+/CD/8+ in group C had no significant changes before and after treatment (P ＞ 0.05 ), but there were significant changes in group T[(12.20 ± 3.98) g/L vs.(6.18± 1.45) g/L, (1.89 ±0.58) g/L vs. (0.63 ±0.15) g/L, (1.29 ± 0.47)g/L vs. (0.51 ± 0.13) g/L, (0.94 ± 0.36) g/L vs.(0.58 ± 0.20) g/L, (0.28 ± 0.06) g/L vs. (0.11 ± 0.04) g/L,(63.11 ± 9.43 )％ vs. (53.26 ± 9.08 )％, ( 38.21 ± 6.15 )％ vs. ( 31.56± 6.02 )％, 1.48 ± 0.37 vs. 1.25 ± 0.43](P＜ 0.05), and there were significant differences between group T and group C after treatment (P＜ 0.05).The ratio of infection in group T was decreased obviously [46.7％ (14/30) vs. 133.3％ (40/30), P ＜ 0.05], but there was no significant difference in group C [126.7％(38/30) vs. 136.7％(41/30),P＞ 0.05]. Conclusion High-flux hemodialysis can improve the immune state and the nuuitional state in MHD patients, and it could decrease the infection rate.

Objective To explore the effects of minimally invasive removal of intracranial hematoma on blood-brain barrier (BBB) index, serum myelin basic protein (MBP) and activity of daily living (ADL) in hypertensive patients with cerebral hemorrhage.Methods Through observing 30cases operated within 3.0 hours, 32 case operated between 3. 1-8. 0 hours, 28 cases operated between 8. 1 to 24.0 hours and 22 cases operated over 24 hours, the changes of BBB index, serum MBP and ADL were analyzed. Results The BBB index and serum MBP were significantly lower in patients operated within 8. 0 hours than in patients operated over 8. 1 hours [≤3.0 hours group:(6.57±0.69)×10-3 and (3. 12±0.40)μg/L;3. 1-8.0 hours group: (7. 37±1.29)×10-3 and (3.25±0.60)μg/L;8. 1-2.0 hours group: ( 12. 02± 1.51 ) × 10 3 and (4. 60±0. 48)μg/L;over 24.0 hours group: ( 14. 68±2.07)×10-3 and (5.88±0.64)μg/L,Q＞13.8,P＜0. 05]. And the ADL was lower in patients operated within 8. 0 hours than in patients operated over 8. 1 hours [≤3.0 hours group: (2. 60± 1.07)scores; 3.1-8.0 hours group: (3. 06±0. 91 )scores;8. 1-24.0 hours group: (4.00±0.67) scores;over 24.0 hours group:(3.68±1.32)scores,Q＞3. 1,P＜0.05].Conclusions The minimally invasive surgery of intracranial hematoma within 8.0 hours can mitigate the cytotoxicity-damaged BBB so as to lighten brain edema and improve the patients quality of life.

Objective To explore the imaging manifestations of low-grade central osteosarcoma (LGCOS) and discuss their pathological features.Methods Twelve patients of LGCOS proved by surgery and pathology were analyzed retrospectively and a review of related literature was performed.All twelve patients had plain X-ray,1 1 patients CT examination,and 10 patients contrast-enhanced MR scan.Imaging features of the LGCOS were summarized,their clinical and pathological manifestations were discussed for differential diagnosis.Their prognosis was evaluated with followed up examination.Results Of the 12patients with LGCOS,six tumors were located in the distal femur,3 in the proximal tibia,2 in the proximal femur and 1 in the talus.The radiographic features of LGCOS were variable.There were 7 patients with predominantly osteolytic destruction,3 patients with mixed sclerotic and lyric changes,with well-defined margins,2 patients with Osteogenic changes on X-ray.On CT,9 patients showed a clear cortical breach,5 patients with soft tissue involvement,6 patients with peripheral incompletely sclerotic zone,2 patients with periosteal reaction.On MRI,there were 10 patients with abnormal signal in medullary cavity,8 patients with soft tissue masses,and all 10 patients exhibited contrast enhancement.The microscopic features of LGCOS were characteristically bland,comprising spindle cells arranged in interlacing fascicles in a heavily collagenous background with variable bone or osteoid production.There were mild nuclear atypia and rare mitoses.In four patients,misdiagnoses were made by biopsy or surgical pathology as fibrous dysplasia or fibrous histiocytoma and other benign lesions,all four patients had postoperative recurrence.Conclusions LGCOS should be differentiated form fibrous dysplasia,non-ossifying fibroma,and other benign lesions.An accurate diagnosis can be made in most cases by careful pathological and radiological correlation.

Microsatellite instability (MSI) is a key biomarker for cancer therapy and prognosis. Tra-ditional experimental assays are laborious and time-consuming, and next-generation sequencing-based computational methods do not work on leukemia samples, paraffin-embedded samples, or patient-derived xenografts/organoids, due to the requirement of matched normal samples. Herein, we developed MSIsensor-pro, an open-source single sample MSI scoring method for research and clinical applications. MSIsensor-pro introduces a multinomial distribution model to quantify poly-merase slippages for each tumor sample and a discriminative site selection method to enable MSI detection without matched normal samples. We demonstrate that MSIsensor-pro is an ultrafast, accurate, and robust MSI calling method. Using samples with various sequencing depths and tumor purities, MSIsensor-pro significantly outperformed the current leading methods in both accuracyand computational cost. MSIsensor-pro is available at https://github.com/xjtu-omics/msisensor-pro and free for non-commercial use, while a commercial license is provided upon request.

Objective:To explore the correlation between muscle CT measurement parameters, energy expenditure and acute exacerbation in patients with stable chronic obstructive pulmonary disease (COPD).Methods:The clinical data of 146 patients with stable COPD from March 2020 to November 2021 in Lu′an Hospital Affiliated to Anhui Medical University (Lu′an People′s Hospital) were retrospectively analyzed. The clinical data were recorded; the lung function was measured by bronchodilator test. The cross-sectional area and CT value of the pectoral muscle were measured by reconstructed CT images of the mediastinum; the total energy consumption was calculated by Weir formula. Acute exacerbations within 3 and 12 months were recorded. Multivariate Logistic regression was used to analyze the independent risk factors for acute exacerbation in patients with stable COPD. The receiver operating characteristic (ROC) curve was used to analyze the efficacy of total energy expenditure, pectoral muscle cross-sectional area and pectoral muscle CT value for predicting acute exacerbation in patients with stable COPD.Results:Among 146 patients with stable COPD, 38 cases (26.03%) developed acute exacerbation within 3 months (acute exacerbation group), and 108 cases (73.97%) did not develop acute exacerbation (non-acute exacerbation group). The proportion of age<60 years old, rate of acute exacerbation within 12 months and rate of pulmonary function grading Ⅲ to Ⅳ in acute exacerbation group were significantly higher than those in non-acute exacerbation group: 71.05% (27/38) vs. 47.22% (51/108), 52.63% (20/38) vs. 30.56% (33/108) and 63.16% (24/38) vs. 37.96% (41/108), the total energy consumption, pectoral muscle cross-sectional area and pectoral muscle CT value were significantly lower than those in non-acute exacerbation group: (2 036.28 ± 163.13) J/d vs. (2 389.59 ± 204.71) J/d, (28.79 ± 3.45) cm 2 vs. (31.61 ± 4.56) cm 2 and (29.79 ± 3.06) HU vs. (34.52 ± 4.38) HU, and there were statistical differences ( P<0.05 or <0.01). Multivariate Logistic regression analysis result showed that age ≥60 years old, lower total energy expenditure, smaller pectoral muscle cross-sectional area and lower pectoral muscle CT value were independent risk factors for acute exacerbation in patients with stable COPD ( OR = 26.493, 1.015, 1.245 and 1.437; 95% CI 3.745 to 187.405, 1.008 to 1.022, 1.002 to 1.546 and 1.109 to 1.861; P<0.01 or <0.05). The ROC curve analysis result showed that combined prediction of the total energy consumption, pectoral muscle cross-sectional area and pectoral muscle CT value for acute exacerbation in patients with stable COPD had the largest area under the curve (0.962), with a sensitivity of 86.1%, a specificity of 80.8%, and the optimal cutoff values of 2 206.12 J/d, 32.39 cm 2 and 31.63 HU, respectively. Conclusions:The elderly age, smaller pectoral muscle cross-sectional area, lower pectoral muscle CT value and lower total energy expenditure are independent risk factors for acute exacerbation in patients with stable COPD. The combination of pectoral muscle cross-sectional area, pectoral muscle CT value and total energy expenditure has a good predictive effect on the risk of acute exacerbation in patients with stable COPD, and relevant indexes can be paid attention to in clinical treatment.

Gliosarcoma (GS) is a rare variant (2%) of glioblastoma (GBM) that poses clinical genomic challenges because of its poor prognosis and limited genomic information. To gain a comprehensive view of the genomic alterations in GS and to understand the molecular etiology of GS, we applied whole-exome sequencing analyses for 28 GS cases (6 blood-matched fresh-frozen tissues for the discovery set, 22 formalin-fixed paraffin-embedded tissues for the validation set) and copy-number variation microarrays for 5 blood-matched fresh-frozen tissues. TP53 mutations were more prevalent in the GS cases (20/28, 70%) compared to the GBM cases (29/90, 32%), and the GS patients with TP53 mutations showed a significantly shorter survival (multivariate Cox analysis, hazard ratio=23.9, 95% confidence interval, 2.87–199.63, P=0.003). A pathway analysis showed recurrent alterations in MAPK signaling (EGFR, RASGRF2 and TP53), phosphatidylinositol/calcium signaling (CACNA1s, PLCs and ITPRs) and focal adhesion/tight junction (PTEN and PAK3) pathways. Genomic profiling of the matched recurrent GS cases detected the occurrence of TP53 mutations in two recurrent GS cases, which suggests that TP53 mutations play a role in treatment resistance. Functionally, we found that TP53 mutations are associated with the epithelial–mesenchymal transition (EMT) process of sarcomatous components of GS. We provide the first comprehensive genome-wide genetic alternation profiling of GS, which suggests novel prognostic subgroups in GS patients based on their TP53 mutation status and provides new insight in the pathogenesis and targeted treatment of GS.
Glioblastoma ; Gliosarcoma* ; Humans ; Prevalence* ; Prognosis

We aimed to develop a whole-genome sequencing(WGS)-based copy number variant(CNV)calling algorithm with the potential of replacing chromosomal microarray assay(CMA)for clinical diagnosis.JAX-CNV is thus developed for CNV detection from WGS data.The perfor-mance of this CNV calling algorithm was evaluated in a blinded manner on 31 samples and com-pared to the 112 CNVs reported by clinically validated CMAs for these 31 samples.The result showed that JAX-CNV recalled 100％of these CNVs.Besides,JAX-CNV identified an average of 30 CNVs per individual,representing an approximately seven-fold increase compared to calls of clinically validated CMAs.Experimental validation of 24 randomly selected CNVs showed one false positive,i.e.,a false discovery rate(FDR)of 4.17％.A robustness test on lower-coverage data revealed a 100％sensitivity for CNVs larger than 300 kb(the current threshold for College of American Pathologists)down to 10×coverage.For CNVs larger than 50 kb,sensi-tivities were 100％for coverages deeper than 20×,97％for 15×,and 95％for 10×.We developed a WGS-based CNV pipeline,including this newly developed CNV caller JAX-CNV,and found it capable of detecting CMA-reported CNVs at a sensitivity of 100％with about a FDR of 4％.We propose that JAX-CNV could be further examined in a multi-institutional study to justify the transition of first-tier genetic testing from CMAs to WGS.JAX-CNV is available at https://github.com/The J acksonLaboratory/JAX-CNV.

Complex structural variants(CSVs)are genomic alterations that have more than two breakpoints and are considered as the simultaneous occurrence of simple structural variants.How-ever,detecting the compounded mutational signals of CSVs is challenging through a commonly used model-match strategy.As a result,there has been limited progress for CSV discovery com-pared with simple structural variants.Here,we systematically analyzed the multi-breakpoint con-nection feature of CSVs,and proposed Mako,utilizing a bottom-up guided model-free strategy,to detect CSVs from paired-end short-read sequencing.Specifically,we implemented a graph-based pattern growth approach,where the graph depicts potential breakpoint connections,and pattern growth enables CSV detection without pre-defined models.Comprehensive evaluations on both simulated and real datasets revealed that Mako outperformed other algorithms.Notably,validation rates of CSVs on real data based on experimental and computational validations as well as manual inspections are around 70％,where the medians of experimental and computational breakpoint shift are 13 bp and 26 bp,respectively.Moreover,the Mako CSV subgraph effectively characterized the breakpoint connections of a CSV event and uncovered a total of 15 CSV types,including two novel types of adjacent segment swap and tandem dispersed duplication.Further analysis of these CSVs also revealed the impact of sequence homology on the formation of CSVs.Mako is publicly available at https://github.com/xjtu-omics/Mako.