[Clinical analysis of 18 children with aggressive mature B-cell lymphoma after liver transplantation].

Objective: To summarize the clinical characteristics, prognostic factors and treatment outcomes of childhood aggressive mature B-cell lymphoma after liver transplantation. Methods: This retrospective study included 18 children with newly diagnosed aggressive mature B-cell lymphoma after liver transplantation and treated from June 2018 to June 2022 in the Department of Hematology and Oncology of Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine. Clinical characteristics, treatment and outcomes of patients at last evaluation were analyzed. Overall survival (OS) and event free survival (EFS) rates were calculated by Kaplan-Meier method and Log-Rank analysis was performed to find factors of poor prognosis. Results: Among all 18 patients, there were 6 males and 12 females, and the age of onset was 40 (35, 54) months. The interval from transplant to tumor diagnosis was 21 (17, 35) months and 5 patients had early onset disease (<1 year since transplant). Seventeen patients had abdominal lesions. Diarrhea, vomiting and abdominal masses were the main clinical manifestations. All patients were Epstein-Barr virus (EBV) related posttransplant lymphoproliferative disorders (PTLD). One patient received individualized therapy due to critical sick at diagnosis, and the remaining 17 patients received CP (cyclophosphamide, methylprednisolone plus rituximab) and (or) modified EPOCH (prednisone, etoposide, doxorubicin, vincristine, cyclophosphamide plus rituximab) regimens. Of all 18 patients, 15 cases got complete response, 2 cases got partial response, 1 patient died of severe infection. The 2-year OS and EFS rates of 18 patients were (94±5)% and (83±8)%, respectively. None of age, gender or early onset disease had effect on OS and EFS rates in univariate analysis (all P>0.05). Conclusions: The symptoms of PTLD were atypical. Close surveillance of EBV-DNA for patients after liver transplantation was crucial to early stage PTLD diagnosis. CP or modified EPOCH regimen was efficient for pediatric patients with aggressive mature B cell lymphoma after liver transplantation.

Warmer temperature during asexual reproduction induce methylome, transcriptomic, and lasting phenotypic changes in Fragaria vesca ecotypes.

Plants must adapt with increasing speed to global warming to maintain their fitness. One rapid adaptation mechanism is epigenetic memory, which may provide organisms sufficient time to adapt to climate change. We studied how the perennial Fragaria vesca adapted to warmer temperatures (28°C vs. 18°C) over three asexual generations. Differences in flowering time, stolon number, and petiole length were induced by warmer temperature in one or more ecotypes after three asexual generations and persisted in a common garden environment. Induced methylome changes differed between the four ecotypes from Norway, Iceland, Italy, and Spain, but shared methylome responses were also identified. Most differentially methylated regions (DMRs) occurred in the CHG context, and most CHG and CHH DMRs were hypermethylated at the warmer temperature. In eight CHG DMR peaks, a highly similar methylation pattern could be observed between ecotypes. On average, 13% of the differentially methylated genes between ecotypes also showed a temperature-induced change in gene expression. We observed ecotype-specific methylation and expression patterns for genes related to gibberellin metabolism, flowering time, and epigenetic mechanisms. Furthermore, we observed a negative correlation with gene expression when repetitive elements were found near (±2 kb) or inside genes. In conclusion, lasting phenotypic changes indicative of an epigenetic memory were induced by warmer temperature and were accompanied by changes in DNA methylation patterns. Both shared methylation patterns and transcriptome differences between F. vesca accessions were observed, indicating that DNA methylation may be involved in both general and ecotype-specific phenotypic variation.

Single-cell analysis of the nervous system at small and large scales with instant partitions.

Single-cell RNA sequencing is a new frontier across all biology, particularly in neuroscience. While powerful for answering numerous neuroscience questions, limitations in sample input size, and initial capital outlay can exclude some researchers from its application. Here, we tested a recently introduced method for scRNAseq across diverse scales and neuroscience experiments. We benchmarked against a major current scRNAseq technology and found that PIPseq performed similarly, in line with earlier benchmarking data. Across dozens of samples, PIPseq recovered many brain cell types at small and large scales (1,000-100,000 cells/sample) and was able to detect differentially expressed genes in an inflammation paradigm. Similarly, PIPseq could detect expected and new differentially expressed genes in a brain single cell suspension from a knockout mouse model; it could also detect rare, virally-la-belled cells following lentiviral targeting and gene knockdown. Finally, we used PIPseq to investigate gene expression in a nontraditional model species, the little skate (Leucoraja erinacea). In total, PIPSeq was able to detect single-cell gene expression changes across models and species, with an added benefit of large scale capture and sequencing of each sample.

[Analysis of 42 cases of childhood superior vena cava syndrome associated with mediastinal malignancy].

Objective: To summarize the clinical features, management and outcome of superior vena cava syndrome (SVCS) associated with mediastinal malignancy in children. Methods: Clinical data of 42 children of SVSC associated with mediastinal malignancy in Shanghai Children's Medical Center from January 2015 to December 2021 were collected and analyzed retrospectively. The clinical manifestations, pathological diagnosis, disease diagnosis process, and prognosis were summarized. Results: Among 42 children of SVCS associated with mediastinal malignancy, there were 31 males and 11 females. The age at diagnosis was 8.5 (1.9, 14.9) years. Cough and wheezing (33 cases, 79%), orthopnea (19 cases, 45%) and facial edema (18 cases, 43%) occurred most commonly. T-cell lymphoblastic lymphoma (T-LBL) was the most frequent pathological diagnosis (25 cases, 60%), followed by T-cell acute lymphoblastic leukemia (T-ALL) (7 cases, 17%), anaplastic large cell lymphoma (4 cases, 10%) and diffuse large B-cell lymphoma (2 cases, 5%), peripheral T-lymphoma, Hodgkin lymphoma, Ewing's sarcoma and germ cell tumor (1 case each). Pathological diagnosis was confirmed by bone marrow aspiration or thoracentesis in 14 cases, peripheral lymph node biopsy in 6 cases, and mediastinal biopsy in 22 cases. Twenty-seven cases (64%) had local anesthesia. Respiratory complications due to mediastinal mass developed in 3 of 15 cases who received general anesthesia. Of the 42 cases, 27 cases had sustained remission, 1 case survived with second-line therapy after recurrence, and 14 cases died (2 cases died of perioperative complications and 12 cases died of recurrence or progression of primary disease). The follow-up time was 36.7 (1.2, 76.1) months for 27 cases in continuous complete remission. The 3-year overall survival (OS) and events free survival (EFS) rates of 42 children were 59% (95%CI 44%-79%) and 58% (95%CI 44%-77%) respectively. Conclusions: SVCS associated with mediastinal malignancy in children is a life-threatening tumor emergency with high mortality. The most common primary disease is T-LBL. The most common clinical symptoms and signs are cough, wheezing, orthopnea and facial edema. Clinical management should be based on the premise of stable critical condition and confirm the pathological diagnosis through minimal invasive operation.

[Multicenter clinical study on the diagnosis and treatment of childhood renal tumor].

Objective: To summarize the effect of Chinese Children's Cancer Group (CCCG) Wilms tumor (WT)-2015 protocol. Methods: This was a prospective study. CCCG-WT-2015 protocol was revised on the basis of the CCCG-WT-2009 protocol. Clinical data of 288 children diagnosed with newly diagnosed kidney neoplasms in fourteen pediatric centers between September 2015 to December 2018 were summarized. The age of onset, distribution of pathological subtypes, staging, curative effect and prognostic factors of these children were analyzed. Kaplan-Meier method was used for survival curve and Log-Rank method was used for univariate analysis. Results: Among 288 cases with kidney neoplasms, there were 261 cases of WT, including 254 cases (97.3%) with favorable histology (FH) WT and 7 cases (2.7%) with unfavorable histology WT (UFHWT). The 3 year events free survival (EFS) rate for FHWT and UFHWT were (88.9±2.1)% and (80.0±17.9)%, which were better than that in WT-2009 (81.2% and 71.7%). In the 96 cases of stage Ⅲ/Ⅳ FHWT with indications for radiotherapy, 76 cases received radiation, another 20 cases received M protocol chemotherapy (cyclophosphamide, etoposide, gentamycin, vincristine and adriamycin) instead of radiation. The 3 year EFS rate for these two groups were (84.7±4.3)% and (84.7±8.1)%(χ2=0.015, P=0.902). There were 22 renal clear cell sarcoma and 5 malignant rhabdoid tumor, 3 year EFS rate of them was (94.4±5.4)% and (20.0±17.9)%. Univariate analysis was performed for age, gender, pathological type, stage, whether rupture occurred during operation, whether complete remission (CR) occurred at the end of treatment and radiotherapy. Pathological types (χ2=44.329,P<0.01) and failure to achieve CR at the end of the treatment (χ2=49.459,P<0.01) were independent factor for predicting survival. Conclusion: Compared with CCCG-WT-2009, treatment of renal tumors in CCCG-WT-2015 study yielded good survival outcome, which can be further applied.

Low-Carbohydrate Diet Score and Coronary Artery Calcium Progression: Results From the CARDIA Study.

OBJECTIVE: To investigate whether low-carbohydrate diets (LCDs) were associated with coronary artery calcium (CAC) progression. Approach and Results: We included the participants who completed computed tomography assessment of baseline CAC in 2000 to 2001 (year 15) and follow-up (year 20 or 25) and food frequency questionnaire (years 0, 7, and 20) in the CARDIA study (Coronary Artery Risk Development in Young Adults). CAC progression was defined as CAC >0 at follow-up among participants with baseline CAC of 0 and an annualized change of 10 or percent change of ≥10% for those with 0

[Performance of combined liquid based cytology and HPV nucleic acid test for detecting cervical precancer among women attending screening].

Objective: To evaluate the clinical performance of HPV genotyping with cytology for detecting cervical precancer among women attending co-testing. Methods: A total of 2 883 females who participated in cervical cancer screening program were recruited from Erdos in 2016. All the participants were tested by cytology and HPV genotyping. In 2017, women with abnormal cytology results or HPV positive were followed up. Pathological cervical intraepithelial neoplasia (CIN) 2+ was the study end-point. Clinical performance indexes were calculated, including sensitivity, specificity, positive predictive value, negative predictive value, referral rate and missed cases. Results: INNO-LiPA resulted in a detection rate of 18.87%(544/2 883) for the 14-type high risk HPV. HPV16 was the most common infectious genotype (4.06%), followed by HPV52 (3.61%), HPV51 (2.50%), HPV58 (1.98%), and HPV18 (1.56%). With more HPV genotypes added into the group, sensitivity increased and the specificity decreased. Addition of HPV16, 58, 33, 39, 52, 18, 31 for detection lead to the maximun value of area under the curve (AUC)=0.913 (95%CI: 0.882-0.944). Compared with traditional screening method by cytology, cotesting decreased the number of missed diagnosis. Meanwhile, the fifth method (co-testing: triage of women with HPV16/18+ , cytological minor abnormalities and HPV58, 33, 39, 52, 31+ or cytological high grade abnormalities) did not increase referral rate (8.99% vs. 8.71%, P=0.525), with five cases of missed diagnosis (sensitivity of 92.1% and specificity of 93.2%). Conclusions: Co-testing with triage of women with HPV16/18+ , cytological minor abnormalities and HPV58, 33, 39, 52, 31+ or cytological high grade abnormalities would provide better clinical performance. In co-testing, triage of HPV16/18 was used in women with normal cytology; triage of HPV58, 33, 39, 52 and 31 was used in women with low-grade abnormal cytology; referral colposcopy was used in women with high-grade abnormal cytology, which would provide better clinical performance.

Z-Score transformation of ADC values: A way to universal cut off between malignant and benign lymph nodes.

PURPOSE: To determine the possibility of a universal cut off value between benign and malignant lymph nodes in patients with tumour by Z-Score transformation method. MATERIALS AND METHODS: Diffusion weighted imaging, ADC measurements of malignant and benign lymph nodes of 6 studies (4 body parts), conducted for 5 times, in two institutions with variable technical details were analyzed in their original value as well as the standardized Z-Score value. The standardized Z-Score value was obtained by subtracting the population mean of the control group from an individual raw score and then dividing the difference by the population standard deviation of the control group. General cut off values were obtained by both Mega-analysis by receiver operator characteristic curve analysis, when data from the 6 studies were combined and Meta-analysis with weighting coefficients and cut off values of the six individual studies. Sensitivity, specificity and accuracy with cut offs from individual studies, meta-analysis and mega-analysis were calculated. Kappa test was performed to assess the consistency of diagnostic test accuracy, between optimized cut offs of individual studies and the proposed universal cut offs obtained from meta-analysis and mega-analysis. RESULTS: The ADC values of benign and malignant lymph nodes are significantly different, but with large overlap across the studies. The overlap can be minimized by Z-Score transformation. The result of ROC analysis of the collective Z-Score transformed ADC values of 6 studies was superior to that of the collective original ADC values (sensitivity: 87.4% versus 67.2%, specificity: 90.5% versus 87.9%, accuracy: 89.6% versus 81.4%). The universal Z-Score cut off from Meta-analysis is also better than the original ADC cut off (sensitivity: 82.8% versus 76.3%, specificity 92.6% versus 62.9%, accuracy 89.6% versus 67.1%). Applied to the individual studies, the universal transformed Z-Score cut offs produced superior consistency with the individual optimal cut offs (individual and meta Z-Score: 0.7228-0.9793; individual and mega Z-Score: 0.7111-0.9169) compared with the universal original ADC cut offs (individual and meta ADC: 0.3030-1.0000; individual and mega ADC 0.3268-0.9618). CONCLUSION: Z-Score transformation could minimize inter-study variations due to heterogeneity of MR systems and sequence parameters, and provide a more consistent universal cut off value between benign and malignant nodes across studies.

Low-level laser irradiation promotes the differentiation of bone marrow stromal cells into osteoblasts through the APN/Wnt/ß-catenin pathway.

OBJECTIVE: The relationship between adiponectin (APN) pathway and Wnt pathway was explored through BMSCs, and the effect of low-level laser irradiation (LLLI) on bone marrow stromal cells (BMSCs) and its mechanism were further studied. MATERIALS AND METHODS: 3-week-old Sprague-Dawley (SD) rats were selected, and mesenchymal stem cells were separately cultured and purified. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to analyze cell proliferation. After osteogenic and adipogenic induction, cultures were conducted, respectively, cells were stained with alizarin red and oil red O. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expressions of osteogenesis-related genes, runt-related transcription factor 2 (RUNX2), and osteocalcin (OC) and those of adipogenesis-related genes, peroxisome proliferator-activated receptor-gamma (PPARγ) and CCAAT/enhancer-binding protein alpha (c/EBPα). Western blotting was used to detect the expressions of ß-catenin in the cytoplasm and nucleus. The lentiviral expression vector of adiponectin receptors (APN-R) was constructed, and the expression of APN receptor genes was silenced. The expressions of ß-catenin in APN receptors and the nucleus within cells were detected. RESULTS: LLLI promoted the bone formation by inducing the differentiation direction of mesenchymal stem cells, increasing the number of osteoblasts in the bone marrow and inhibiting the reduction of the number of adipocytes. LLLI regulates the Wnt pathway, promotes the entry of ß-catenin into the nucleus, activates the osteogenic effect of the Wnt pathway so as to promote the bone formation of osteoblasts and inhibit bone resorption of osteoclasts. LLLI promotes the entry of ß-catenin into the nucleus and the osteogenic differentiation of BMSCs through the APN pathway. CONCLUSIONS: In summary, LLLI can promote osteogenesis and inhibit adipocytes formation, thus attenuating bone resorption of osteoclasts. The mechanism of LLLI is that it promotes the entry of ß-catenin into the nucleus and regulates the Wnt pathway and the differentiation direction of mesenchymal stem cells through the APN signal pathway, thus promoting bone formation.

The importance of cerebellar cortex and facial nucleus in acquisition and retention of eyeblink/NM conditioning: evidence for critical unilateral regulation of the conditioned response.

Experiment 1 examined acquisition of the classically conditioned eyeblink response in rabbits (Oryctolagus cuniculus) during reversible cooling lesions of the cerebellar cortex (CX), facial nucleus (FN), or lateral parvocellular reticular formation (RF). Retention was then evaluated during periods of training during reversible cooling lesions and without cooling in rabbits that had acquired the conditioned response. We found that cooling the CX did not prevent acquisition, but did retard the acquisition rate. Cooling the FN during acquisition prevented the expression of the unconditioned and conditioned response, but did not prevent the acquisition when assessed during subsequent training without cooling. Cooling the RF had no effect on the acquisition or expression of the conditioned response. During subsequent retention testing, in well-trained animals, cooling the CX did not abolish the learned response. Cooling the FN abolished both the conditioned and the unconditioned response. The results from Experiment 1 indicate that the CX is more important for acquisition than retention of the conditioned response. The FN is not important for the acquisition of the conditioned response, but is essential for the expression of the conditioned and unconditioned response. Experiment 2 examined bilateral recordings from the cerebellum in well-trained rabbits, before and during interpositus or FN cooling. We found that cooling the interpositus abolished all learning related activity in the ipsilateral or contralateral cerebellum, but did not affect the stimulus evoked responses. Cooling the FN did not abolish stimulus evoked activity or learning related activity in the cerebellum. The results emphasize the critical importance of the ipsilateral cerebellum in classical eyeblink conditioning and suggest that the memory trace for this type of learning is mediated by unilateral circuitry.

Unilateral inferior olive NMDA lesion leads to unilateral deficit in acquisition and retention of eyelid classical conditioning.

New Zealand White rabbits (Oryctolagus cuniculus) were trained for acquisition (N = 21) or retention (N = 10) of classical eyelid conditioning with unilateral or bilateral N-methyl-DL-aspartate chemical lesions of the rostromedial dorsal accessory inferior olive (rmDAO; multiple injections totaling 76 to 342 nmol). In all instances, subjects were unable to learn or retain conditioning on the side contralateral to the lesion. Learning rates were comparable for lesions outside of the rmDAO and sham operates. These findings demonstrate a specific unilateral deficit whereas in previous research the answer to this question was ambiguous since electrolytic lesions effectively cause bilateral olivary lesions. This research agrees with the concept that the inferior olive projects essential information about the unconditioned stimulus to a cerebellar locus of learning and memory for classical conditioning.

Reversible lesions of the cerebellar interpositus nucleus during acquisition and retention of a classically conditioned behavior.

Previous lesion, recording, and stimulation studies have implicated the cerebellum and its associated circuitry as essentially involved in classical conditioning of discrete, somatic muscle responses. In 2 experiments, the interpositus nucleus of the cerebellum was assessed for the plasticity associated with learning and memory of the nictitating membrane (NM) response by using local cooling as a reversible lesion technique. In well-trained animals (Experiment 1), NM conditioned responses (CRs) were abolished during cooling of the interpositus but reappeared when the interpositus returned to body temperature. This cooling-warming protocol could be repeated many times. Cooling could be prolonged (one session, approximately 1 hr) with recovery of NM CRs as tested on the next day. Multiple-unit recordings related to learning were also absent in the interpositus and red nucleus during cooling. In naive animals (Experiment 2), both behavioral and unit CRs did not develop while training with cooling. There was no evidence of savings when training continued without cooling: Behavioral and unit CRs developed as if the animals were still naive. These results support the idea that the interpositus nucleus of the cerebellum is the critical locus for learning and memory of this classical CR.