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Aim: To retrospectively explored whether systematic training in the use of Liver Imaging Reporting and Data System (LI-RADS) v2018 on computed tomography (CT) can improve the interobserver agreements and performances in LR categorization for focal liver lesions (FLLs) among different radiologists., Materials and Methods: A total of 18 visiting radiologists and the liver multiphase CT images of 70 hepatic observations in 63 patients at high risk of HCC were included in this study. The LI-RADS v2018 training procedure included three thematic lectures, with an interval of 1 month. After each seminar, the radiologists had 1 month to adopt the algorithm into their daily work. The interobserver agreements and performances in LR categorization for FLLs among the radiologists before and after training were compared., Results: After training, the interobserver agreements in classifying the LR categories for all radiologists were significantly increased for most LR categories (P < 0.001), except for LR-1 (P = 0.053). After systematic training, the areas under the curve (AUCs) for LR categorization performance for all participants were significantly increased for most LR categories (P < 0.001), except for LR-1 (P = 0.062)., Conclusion: Systematic training in the use of the LI-RADS can improve the interobserver agreements and performances in LR categorization for FLLs among radiologists with different levels of experience., (© 2024. The Author(s) under exclusive licence to Japan Radiological Society.)

Deletion and missense or nonsense mutation of RAB39B gene cause familial Parkinson's disease (PD). We hypothesized that deletion and mutation of RAB39B gene induce degeneration of dopaminergic neurons by decreasing protein level of functional RAB39B and causing RAB39B deficiency. Cellular model of deletion or mutation of RAB39B gene-induced PD was prepared by knocking down endogenous RAB39B in human SH-SY5Y dopaminergic cells. Transfection of shRNA-induced 90% reduction in RAB39B level significantly decreased viability of SH-SY5Y dopaminergic neurons. Deficiency of RAB39B caused impairment of macroautophagy/autophagy, which led to increased protein levels of α-synuclein and phospho-α-synuclein Ser129 within endoplasmic reticulum (ER) and mitochondria. RAB39B deficiency-induced increase of ER α-synuclein and phospho-α-synuclein Ser129 caused activation of ER stress, unfolded protein response, and ER stress-induced pro-apoptotic cascade. Deficiency of RAB39B-induced increase of mitochondrial α-synuclein decreased mitochondrial membrane potential and increased mitochondrial superoxide. RAB39B deficiency-induced activation of ER stress pro-apoptotic pathway, mitochondrial dysfunction, and oxidative stress caused apoptotic death of SH-SY5Y dopaminergic cells by activating mitochondrial apoptotic cascade. In contrast to neuroprotective effect of wild-type RAB39B, PD mutant (T168K), (W186X), or (G192R) RAB39B did not prevent tunicamycin- or rotenone-induced increase of neurotoxic α-synuclein and activation of pro-apoptotic pathway. Our results suggest that RAB39B is required for survival and macroautophagy function of dopaminergic neurons and that deletion or PD mutation of RAB39B gene-induced RAB39B deficiency induces apoptotic death of dopaminergic neurons via impairing autophagy function and upregulating α-synuclein., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Objectives: Allogeneic hematopoietic cell transplantation (HCT) is associated with acute graft-vs.-host disease (aGVHD). The presented study applied a novel multiplex antibody-based proximity extension assay (PEA) proteomic platform that can detect thousands of serum proteins simultaneously for the identification of potential biomarkers of aGVHD., Methods: Serum samples from 28 patients who underwent allogeneic HCT for acute myeloid leukemia (AML) were analyzed; 17 were diagnosed with grade II-IV aGVHD while 11 patients were not. Samples collected on day -6, day 0, +14, +30, +60 and +90 post-HCT were analyzed for the relative concentrations of 552 proteins. The concentration of each protein from baseline to the closest time point before onset of aGVHD, or to the latest time point in control patients, was documented., Results: Individualized analysis identified 26 proteins demonstrating ≥3-fold increase at aGVHD onset compared to baseline, eliminating proteins with a similar increase in controls. Another approach used paired t-testing and logistic regression that identified a four-marker panel, including SLAMF7, IL-1ra, BTN3A2 and DAB2, where individual log-likelihood ratios ranged from 3.99 to 8.15 (logistic regression, p=0.004-0.046). When combined, the four-marker panel demonstrated an area under the curve (AUC) of 0.90 (95% CI: 0.78-1.00; p=0.0006) with high negative predictive value of 81.8% and positive predictive value of 86.7%. All four markers play a physiological role in immune regulation. Among these, three were also present in the individualized analysis (SLAMF7, IL-1ra and BTN3A2)., Conclusions: We conclude that serum proteins identified using multiplex proteomics, particularly SLAMF7, IL-1ra, BTN3A2 and DAB2, may potentially predict aGVHD., (© 2023 Walter de Gruyter GmbH, Berlin/Boston.)

Objectives: Ovarian cancer is the most lethal gynecological malignancy in developed countries. One of the key associations with the high mortality rate is diagnosis at late stages. This clinical limitation is primarily due to a lack of distinct symptoms and detection at the early stages. The ovarian cancer biomarker, CA125, is mainly effective for identifying serous ovarian carcinomas, leaving a gap in non-serous ovarian cancer detection. Mucin 13 (MUC13) is a transmembrane, glycosylated protein with aberrant expression in malignancies, including ovarian cancer. We explored the potential of MUC13 to complement CA125 as an ovarian cancer biomarker, by evaluating its ability to discriminate serous and non-serous subtypes of ovarian cancer at FIGO stages I-IV from benign conditions., Methods: We used our newly developed, high sensitivity ELISA to measure MUC13 protein in a large, well-defined cohort of 389 serum samples from patients with ovarian cancer and benign conditions., Results: MUC13 and CA125 serum levels were elevated in malignant compared to benign cases (p<0.0001). Receiver-operating characteristic (ROC) curve analysis showed similar area under the curve (AUC) of 0.74 (MUC13) and 0.76 (CA125). MUC13 concentrations were significantly higher in mucinous adenocarcinomas compared to benign controls (p=0.0005), with AUC of 0.80. MUC13 and CA125 showed significant elevation in early-stage cases (stage I-II) in relation to benign controls (p=0.0012 and p=0.014, respectively)., Conclusions: We report the novel role of MUC13 as a serum ovarian cancer biomarker, where it could complement CA125 for detecting some subtypes of non-serous ovarian carcinoma and early-stage disease., (© 2022 Walter de Gruyter GmbH, Berlin/Boston.)

Background: We aimed to evaluate the value of using preoperative magnetic resonance imaging (MRI) features and clinical indicators to predict the early response of hepatocellular carcinoma (HCC) to transcatheter arterial chemoembolization (TACE). We also aimed to establish a preoperative prediction model., Methods: We retrospectively reviewed data of 111 patients with HCC who underwent magnetic resonance imaging (MRI) before the first TACE and underwent MRI or computed tomography between 30 and 60 days after TACE. We used the modified response evaluation criteria in solid tumors for evaluating the TACE response. We used univariate and multivariate logistic regression analyses to identify independent predictors based on MRI features and clinical indicators. Moreover, receiver operating characteristic (ROC) curve analyses were performed to assess the diagnostic performance of the prediction model and each independent predictor., Results: Among the 111 included patients, 85 were men (76.6%). Patient age was 31-86 years (average age, 61.08 ± 11.50 years). After the first treatment session, 56/111 (50.5%) patients showed an objective response (complete response + partial response), whereas the remaining showed non-response (stable disease + local progressive disease). In the univariate analysis, we identified irregular margins, number of nodules, and satellite nodules as predictors of early objective response. However, in the multivariate logistic regression analysis, irregular margins, number of nodules and pretreatment platelet were identified as the independent predictors of early objective response. A combined prediction model was then established, which factored in irregular margins, the number of nodules, and the pretreatment platelet count. This model showed good diagnostic performance (area under the ROC curve = 0.755), with the sensitivity, specificity, positive predictive value, and negative predictive value being 78.6%, 69.1%, 72.1%, and 76.0%, respectively., Conclusions: Irregular margins, the number of nodules and the pretreatment platelet count are independent predictors of the early response of HCC to TACE. Our clinical combined model can provide a superior predictive power to a single indicator., (© 2022. The Author(s).)

Background: The Liver Imaging Reporting and Data System (LI-RADS) was established for noninvasive diagnosis for hepatocellular carcinoma (HCC). However, whether training can improve readers' agreement with the expert consensus and inter-reader agreement for final categories is still unclear., Purpose: To explore training effectiveness on readers' agreement with the expert consensus and inter-reader agreement., Study Type: Prospective., Subjects: Seventy lesions in 61 patients at risk of HCC undergoing liver MRI; 20 visiting scholars., Field Strength/sequence: 1.5 T or 3 T, Dual-echo T 1 WI, Fast spin-echo T 2 WI, SE-EPI DWI, and Dynamic multiphase fast gradient-echo T 1 WI., Assessment: Seventy lesions assigned LI-RADS categories of LR1-LR5, LR-M, and LR-TIV by three radiologists in consensus were randomly selected, with 10 cases for each category. The consensus opinion was the standard reference. The third radiologist delivered the training. Twenty readers reviewed images independently and assigned each an LI-RADS category both before and after the training., Statistical Tests: Accuracy, sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio, negative likelihood ratio, receiver operating characteristic (ROC) analysis, simple and weighted kappa statistics, and Fleiss kappa statistics., Results: Before and after training: readers' AUC (areas under ROC) for LR-1-LR-5, LR-M, and LR-TIV were 0.898 vs. 0.913, 0.711 vs. 0.876, 0.747 vs. 0.860, 0.724 vs. 0.815, 0.844 vs. 0.895, 0.688 vs. 0.873, and 0.720 vs. 0.948, respectively, and all improved significantly (P < 0.05), except LR-1(P = 0.25). Inter-reader agreement between readers for LR-1-LR-5, LR-M, LR-TIV were 0.725 vs. 0.751, 0.325 vs. 0.607, 0.330 vs. 0.559, 0.284 vs. 0.488, 0.447 vs. 0.648, 0.229 vs. 0.589, and 0.362 vs. 0.852, respectively, and all increased significantly (P < 0.05). For training effectiveness on both AUC and inter-reader agreement, LR-TIV, LR-M, and LR-2 improved most, and LR-1 made the least., Data Conclusion: This study shows LI-RADS training could improve reader agreement with the expert consensus and inter-reader agreement for final categories., Level of Evidence: 2 TECHNICAL EFFICACY STAGE: 2., (© 2021 International Society for Magnetic Resonance in Medicine.)

Background: Traumatic brain injury (TBI) is a significant public health issue affecting nearly 69 million patients worldwide per year. Reliable diagnostic biomarkers are urgently needed to aid in disease diagnosis and prognosis and to guide patient aftercare. Blood biomarkers represent an attractive modality to quickly, cheaply, and objectively evaluate clinical status. We hypothesize that deep and quantitative plasma proteomic profiling with a novel technology, proximity extension assay, may lead to the discovery of diagnostic and/or prognostic biomarkers of TBI., Methods: We used high-throughput proximity extension assays (PEA) to quantify the relative abundance of over 1000 unique proteins in plasma. PEA is a highly sensitive multiplex immunoassay capable of detecting very low-abundance proteins (down to fg/mL) in complex biological matrices. Our patient cohort consisted of severe TBI (sTBI) patients, matched healthy controls, and another non-TBI group that was included in the analysis to validate the specificity of the candidates during the selection process. The obtained protein quantification data was then filtered to identify candidate biomarkers through statistical analysis, literature searches, and comparison to our reference control groups., Results: Overall, we identified 6 novel candidate TBI biomarkers. Candidates exhibit a significant increase in plasma protein abundance in sTBI when comparing between healthy controls and sTBI patients. Candidates generally had low expression in our reference groups compared with the sTBI group., Conclusions: Our preliminary findings represent a starting point for future validation. These biomarkers, either alone or in combination, may have significant clinical utility in aiding in TBI diagnosis, prognosis, and/or management., (© American Association for Clinical Chemistry 2021. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background and Aims: Liver imaging reporting and data system (LI-RADS) provides standardized lexicon and categorization for diagnosing hepatocellular carcinoma (HCC). However, there is limited knowledge about the effect of LI-RADS training. We prospectively explored whether the systematic training of LI-RADS v2018 on magnetic resonance imaging (MRI) can effectively improve the diagnostic performances of different radiologists for HCC., Methods: A total of 20 visiting radiologists and the multiparametric MRI of 70 hepatic observations in 61 patients with high risk of HCC were included in this study. The LI-RADS v2018 training procedure included three times of thematic lectures (each lasting for 2.5 h) given by a professor specialized in imaging diagnosis of liver, with an interval of a month. After each seminar, the radiologists had a month to adopt the algorithm into their daily work. The diagnostic performances and interobserver agreements of these radiologists adopting the algorithm for HCC diagnosis before and after training were compared., Results: A total of 20 radiologists (male/female, 12/8; with an average age of 36.75±4.99 years) were enrolled. After training, the interobserver agreements for the LI-RADS category for all radiologists ( p= 0.005) were increased. The sensitivity, specificity, positive predictive value, negative predictive value, and coincidence rate of all radiologists for HCC diagnosis before and after training were 43% vs. 54%, 86% vs. 88%, 74% vs. 81%, 62% vs. 67%, and 65% vs. 71%, respectively. The diagnostic performances of all radiologists ( p< 0.001) showed improvement after training., Conclusions: The systematic training of LI-RADS can effectively improve the diagnostic performances of radiologists with different experiences for HCC., Competing Interests: The authors have no conflict of interests related to this publication., (© 2021 Authors.)

Probing the human proteome in tissues and biofluids such as plasma is attractive for biomarker and drug target discovery. Recent breakthroughs in multiplex, antibody-based, proteomics technologies now enable the simultaneous quantification of thousands of proteins at as low as sub fg/ml concentrations with remarkable dynamic ranges of up to 10-log. We herein provide a comprehensive guide to the methodologies, performance, technical comparisons, advantages, and disadvantages of established and emerging technologies for the multiplexed ultrasensitive measurement of proteins. Gaining holistic knowledge on these innovations is crucial for choosing the right multiplexed proteomics tool for applications at hand to critically complement traditional proteomics methods. This can bring researchers closer than ever before to elucidating the intricate inner workings and cross talk that spans multitude of proteins in disease mechanisms., Competing Interests: Conflict of interest The authors declare no competing interests., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Despite significant investment of funds and resources, few new cancer biomarkers have been introduced to the clinic in the last few decades. Although many candidates produce promising results in the laboratory, deficiencies in sensitivity, specificity, and predictive value make them less than desirable in a patient setting. This review will analyze these challenges in detail as well as discuss false discovery, problems with reproducibility, and tumor heterogeneity. Circulating tumor DNA (ctDNA), an emerging cancer biomarker, is also analyzed, particularly in the contexts of assay specificity, sensitivity, fragmentation, lead time, mutant allele fraction, and clinical relevance. Emerging artificial intelligence technologies will likely be valuable tools in maximizing the clinical utility of ctDNA which is often found in very small quantities in patients with early-stage tumors. Finally, the implications of challenging false discoveries are examined and some insights about improving cancer biomarker discovery are provided. See all articles in this CEBP Focus section, "NCI Early Detection Research Network: Making Cancer Detection Possible.", (©2020 American Association for Cancer Research.)

Background: We investigated an ultrasensitive prostate-specific antigen (uPSA) immunoassay (MesoScale; lower limit of detection (LLD) of 0.0035 pg/mL) to monitor patients with prostate cancer (PCa) following radical prostatectomy (RP) and to examine whether changes in PSA in the conventionally undetectable range (<1 pg/mL) can predict biochemical relapse (BCR)., Methods: We measured uPSA in serial serum samples (N = 100) collected from 20 RP cases with a third-generation ELISA (LLD of 1 pg/mL) and the fifth-generation MesoScale assay. We analyzed the PSA nadir changes to classify patients into BCR or non-BCR groups, observed the trends in PSA kinetics, and associated BCR status with clinicohistopathological features., Results: The ELISA could quantify PSA in only 38% of the RP samples, detecting BCR in 7 of 20 patients with PCa. The MesoScale assay quantified PSA in all samples, showing 8 of 20 patients with BCR. However, there was no significant difference between the median time to BCR detection based on ELISA (1016 days) compared with MesoScale data (949 days). Gleason scores were higher in the BCR groups compared with non-BCR. There was no significant difference for other clinicohistopathological parameters., Conclusions: The uPSA MesoScale technology could track miniscule changes in serum PSA in the range of 0.003-1 pg/mL in all RP cases. However, PSA kinetics and nadir at concentrations <2 pg/mL fluctuated, and increases below this range could not reliably suggest signs of BCR. Instead, ultrasensitive fifth-generation PSA assays may hold clinical potential for measuring the low concentrations of PSA in women for various medical contexts., (© American Association for Clinical Chemistry 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: To account for cancer heterogeneity, we previously introduced the concept of "personalized" tumor markers, which are biomarkers that are informative in subsets of patients or even a single patient. Recent developments in various multiplex protein technologies create excitement for the discovery of markers of tumor burden in individual patients, but the reliability of the technologies remains to be tested for this purpose. Here, we sought to explore the potential of a novel proteomics platform, which utilizes a multiplexed antibody microarray, to detect changes in serum protein concentration that may correlate to tumor burden in pancreatic cancer. Methods: We applied the Quantibody® Human Kiloplex Array to simultaneously measure 1,000 proteins in sera obtained pre- and post-surgically from five pancreatic cancer patients. We expected that proteins which decreased post-surgery may correlate to tumor burden. Sera from two healthy individuals, split into two aliquots each, were used as controls. To validate the multiplexed results, we used single-target ELISA assays to measure the proteins with the largest serum concentration changes after surgery in sera collected pre- and post-surgically from the previous five patients and 10 additional patients. Results: The multiplexed array revealed nine proteins with more than two-fold post-surgical decrease in at least two of five patients. However, validation using single ELISAs showed that only two proteins tested displayed more than two-fold post-surgical decrease in one of the five original patients. In the independent cohort, six of the proteins tested showed at least a two-fold decrease post-surgery in at least one patient. Conclusions: Our study found that the Quantibody® Human Kiloplex Array results could not be reliably replicated with individual ELISA assays and most hits would likely represent false positives if applied to biomarker discovery. These findings suggest that data from novel, high-throughput proteomic platforms need stringent validation to avoid false discoveries., Competing Interests: No competing interests were disclosed., (Copyright: © 2020 Ren AH et al.)

Background: To account for cancer heterogeneity, we previously introduced the concept of "personalized" tumor markers, which are biomarkers that are informative in subsets of patients or even a single patient. Recent developments in various multiplex protein technologies create excitement for the discovery of markers of tumor burden in individual patients, but the reliability of the technologies remains to be tested for this purpose. Here, we sought to explore the potential of a novel proteomics platform, which utilizes a multiplexed antibody microarray, to detect changes in serum protein concentration that may correlate to tumor burden in pancreatic cancer. Methods: We applied the Quantibody® Human Kiloplex Array to simultaneously measure 1,000 proteins in sera obtained pre- and post-surgically from five pancreatic cancer patients. We expected that proteins which decreased post-surgery may correlate to tumor burden. Sera from two healthy individuals, split into two aliquots each, were used as controls. To validate the multiplexed results, we used single-target ELISA assays to measure the proteins with the largest serum concentration changes after surgery in sera collected pre- and post-surgically from the previous five patients and 10 additional patients. Results: The multiplexed array revealed nine proteins with more than two-fold post-surgical decrease in at least two of five patients. However, validation using single ELISAs showed that only two proteins tested displayed more than two-fold post-surgical decrease in one of the five original patients. In the independent cohort, six of the proteins tested showed at least a two-fold decrease post-surgery in at least one patient. Conclusions: Our study found that the Quantibody® Human Kiloplex Array results could not be reliably replicated with individual ELISA assays and most hits would likely represent false positives if applied to biomarker discovery. These findings suggest that data from novel, high-throughput proteomic platforms need stringent validation to avoid false discoveries., Competing Interests: No competing interests were disclosed., (Copyright: © 2020 Ren AH et al.)

Aim: To investigate the diagnostic performance of Liver Imaging Reporting and Data System (LI-RADS) version 2017 for diagnosing hepatocellular carcinoma (HCC), by using major features only and combined major and ancillary features on computed tomography (CT)., Materials and Methods: A total of 147 HCC, 35 non-HCC malignancy, and 37 benign lesions in 205 patients at high risk of HCC were evaluated retrospectively, and the diagnostic performance of LI-RADS for diagnosing HCC were compared between using major features only and adopting major and ancillary features in combination., Results: When using LR-5 as a predictor for diagnosing HCC, the diagnostic specificity (90.3% versus 91.7%), positive predictive value (92.3% versus 93.3%), and accuracy (68% versus 68.8%) were increased based on major and ancillary features in combination than just using major features on CT. When using LR-4/5 as a predictor for diagnosing HCC, the diagnostic sensitivity (78.9% versus 85.7%), negative predictive value (64.4% versus 72%), and accuracy (78.5% versus 82.2%) were increased while preserving a high specificity (77.8% versus 75%), according to major and ancillary features in combination rather than just using major features on CT. The LI-RADS categories of 8.7% (19/219) lesions were adjusted by adding the ancillary features on CT., Conclusion: Adding the ancillary features visible on CT can improve the diagnostic performance of the LI-RADS v2017 algorithm for diagnosing HCC, especially for LR-3 lesions., (Copyright © 2020. Published by Elsevier Ltd.)

Objectives: Kallikrein-related peptidases (KLKs) are a subgroup of 15 secreted chymotrypsin- and trypsin-like serine proteases that have been reported to possess novel functions in innate immunity and inflammation. Since the potential role of KLKs in immunity has not been studied in detail at the protein level, we examined the expression pattern of 12 members of the KLK family in immune-related tissues., Design & Methods: Protein expression in tissue extracts was evaluated using immunoassays (ELISA). Immunohistochemistry (IHC) was performed on representative sections of tonsil and lymph nodes to determine the cellular localization of the KLK family members., Results: ELISA profiling of KLK3-KLK15 (except KLK12) revealed higher protein levels in the tonsil, compared to the lymph nodes and spleen. Relatively high protein levels in the tonsil were observed for KLK7, KLK9, KLK10 and KLK13. Expression of these KLKs was significantly lower in lymph nodes and spleen. IHC analysis in tonsil unveiled that KLK9 and KLK10 were differentially expressed in lymphoid cells. KLK9 was strongly expressed in the germinal center of lymphoid follicles where activated B-cells reside, whereas KLK10 was expressed in the follicular dendritic cells (FDCs) that are vital for maintaining the cycle of B cell maturation., Conclusion: Overall, our study revealed the possible implications of KLK expression and regulation in the immune cells of lymphoid tissues., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.)

Mass spectrometry-based assays have been increasingly implemented in various disciplines in clinical diagnostic laboratories for their combined advantages in multiplexing capacity and high analytical specificity and sensitivity. It is now routinely used in areas including reference methods development, therapeutic drug monitoring, toxicology, endocrinology, paediatrics, immunology and microbiology to identify and quantify biomolecules in a variety of biological specimens. As new ionisation methods, instrumentation and techniques are continuously being improved and developed, novel mass spectrometry-based clinical applications will emerge for areas such as proteomics, metabolomics, haematology and anatomical pathology. This review will summarise the general principles of mass spectrometry and specifically highlight current and future clinical applications in anatomical pathology., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. No commercial re-use. See rights and permissions. Published by BMJ.)

Human stress recovery response to landscapes is under discussion in Chinese settings. The present study aimed to clarify the stress recovery effects of campus street trees on graduate students in autumn. A total of 150 participants (23.75 ± 1.01 years old) completed the Trier Social Stress Test (TSST) and were then randomly assigned to view one of five virtual environments, including the street trees Sophora japonica , Ginkgo biloba , Platanus acerifolia , Koelreuteria paniculata , and the indoor environment (control). Physiological responses were measured by R-R interval and electroencephalography (EEG). Psychological responses were examined through the state version of the State-Trait Anxiety Inventory (STAI-S) and the Perceived Restorativeness Scale (PRS). Results showed that R-R intervals significantly increased while viewing all street trees. Both alpha and beta brainwave activities while viewing S. japonica and G. biloba were remarkably higher than those while viewing P. acerifolia and K. paniculata . The STAI-S scores significantly decreased, and the positive PRS scores were registered after viewing street trees. We concluded that a brief virtual visual experience of campus street trees in autumn has stress recovery effects on graduate students, and the different levels of stress recovery are associated with different types of street trees., Competing Interests: The authors declare no conflict of interest.

Immune-checkpoint blockade (ICB) uses antibody targeting of specific inhibitory receptors and ligands. The major limitations of ICB, such as high cost, limited success rate, and immune-related adverse events (irAE), highlight the need for predictive biomarkers. We analyzed pre-immunotherapy and post-immunotherapy serum samples of 24 patients treated with pembrolizumab for changes in PD-1 and over 1,000 additional protein markers using a multiplex proximity extension assay (PEA) to identify potential predictive biomarkers of response and/or toxicity. Candidates were selected based on the criteria that at least 2 patients within any of 3 patient groups (responders without irAEs, responders with irAEs, or nonresponders with irAEs) had either a ≥4-fold increase or 4-fold decrease in expression post-immunotherapy. Female and male control samples were used as technical duplicates. A patient group with no response and no irAEs was used to exclude candidates. Following treatment with pembrolizumab, there was a relative increase of PD-1 in the serum of all patients, compared with controls (average 4.4-fold). We identified 7 additional serum proteins that met our candidate selection criteria. These candidate markers did not have any significant association with response or toxicity to pembrolizumab. Overall, we show that serum PD-1 increases post-therapy with pembrolizumab treatment but has no predictive value for response or toxicity in this small set of patients., (©2019 American Association for Cancer Research.)

Background: The Liver Imaging Reporting and Data System (LI-RADS) is widely adopted for noninvasive diagnosis of hepatocellular carcinoma (HCC). It's updated to version 2018 recently, with some major changes compared with v2017. However, the diagnostic performance of LI-RADS v2018 and its difference with v2017 are yet to be validated., Purpose: To compare the diagnostic performances of LI-RADS on MR for diagnosing HCC between v2017 and v2018., Study Type: Retrospective., Subjects: In all, 181 patients with 217 hepatic observations (146 HCCs, 16 non-HCC malignancies and 55 benign lesions) with liver MRI and pathological or follow-up imaging diagnoses., Field Strength/sequence: 1.5 T or 3 T MRI. Dual-echo T 1 WI, T 2 WI, diffusion-weighted imaging, and a liver acquisition with volume acceleration. Assessment Images were independently interpreted by three radiologists, and then in consensus for observations with different LR categories, according to LI-RADS v2017 and v2018, separately., Statistical Tests: Sensitivity, specificity, accuracy, positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (+LR), and Youden index., Results: When adopting LR-5 as a predictor of HCC, the sensitivity (80.8% vs. 71.2%), NPV (69.6% vs. 60.7%), and accuracy (83.9% vs. 77.9%) were all increased for LI-RADS v2018 compared with v2017, with a greater Youden index (0.709 vs. 0.627). However, the diagnostic performances of MRI for diagnosing HCC were not changed while adopting LR-4/5 as a predictor. The threshold growths of 76% (19/25) observations in v2017 were revised to subthreshold growth in v2018, and 16 LR-4 observations in v2017 were changed to LR-5 based on v2018., Data Conclusion: The diagnostic performance of LI-RADS v2018 for diagnosing HCC is superior to v2017, with a greater sensitivity, NPV, and accuracy. The revisions in v2018 mainly affect the categorization when adopting LR-5 as a predictor of HCC., Level of Evidence: 4 Technical Efficacy Stage: 2 J. Magn. Reson. Imaging 2019;50:746-755., (© 2019 International Society for Magnetic Resonance in Medicine.)

The dysfunction of the medial prefrontal cortex is associated with affective disorders and non-motor features in Parkinson's disease. However, the exact role of the mediodorsal thalamic nucleus in the function of the prefrontal cortex remains unclear. To study the possible effects of the mediodorsal thalamic nucleus on the neurological function of the medial prefrontal cortex, a model of Parkinson's disease was established by injecting 8 µg 6-hydroxydopamine into the substantia nigra compacta of rats. After 1 or 3 weeks, 0.3 µg ibotenic acid was injected into the mediodorsal thalamic nucleus of the midbrain. At 3 or 5 weeks after the initial injury, neuronal discharge in medial prefrontal cortex of rat brain was determined electrophysiologically. The numbers of dopamine-positive neurons and tyrosine hydroxylase immunoreactivity in substantia nigra compacta and ventral tegmental area were detected by immunohistochemical staining. Results demonstrated that after injury, the immunoreactivity of dopamine neurons and tyrosine hydroxylase decreased in the substantia nigra compacta and ventral tegmental areas of rats. Compared with normal medial prefrontal cortical neurons, at 3 and 5 weeks after substantia nigra compacta injury, the discharge frequency of pyramidal neurons increased and the discharge pattern of these neurons tended to be a burst-discharge, with an increased discharge interval. The discharge frequency of interneurons decreased and the discharge pattern also tended to be a burst-discharge, but the discharge interval was only higher at 3 weeks. At 3 weeks after the combined lesions, the discharge frequency, discharge pattern and discharge interval were restored to a normal level in pyramidal neurons and interneurons in medial prefrontal cortex. These findings have confirmed that mediodorsal thalamic nucleus is involved in regulating neuronal activities of the medial prefrontal cortex. The changes in the function of the mediodorsal thalamic nucleus may be associated with the abnormal discharge activity of the medial prefrontal cortex neurons after substantia nigra compacta injury. All experimental procedures were approved by the Institutional Animal Care and Use Committee of Xi'an Jiaotong University, China (approval No. XJTULAC2017-067) on August 26, 2017., Competing Interests: None

Objective: This study aims to investigate the effect of t'ai chi exercise on hypertension in young and middle-aged in-service staff., Methods: A total of 208 subjects with grade 1 hypertension were enrolled into this study. These subjects were randomly divided into two groups: research group and control group (n = 104, each). On the basis of general daily lifestyle intervention, subjects in the research group underwent 24-Style Simplified t'ai chi exercise for 3 months, whereas subjects in the control group underwent general daily lifestyle intervention. All subjects were followed up at the first and third month of intervention. The body mass index (BMI), blood pressure, blood lipid, and other indexes were measured before and after the intervention, and quality of life was evaluated., Results: (1) In the research group, after 1 month of exercise, systolic blood pressure (SBP), heart rate (HR), triglyceride (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C) significantly decreased (p < 0.05), while BMI, blood glucose (Glu), diastolic blood pressure (DBP), and pulse pressure (PP) did not significantly change. Furthermore, after 3 months of exercise, BMI, HR, SBP, DBP, PP, TG, TC, LDL-C, and Glu all significantly decreased (p < 0.05). (2) Moreover, the quality of life of subjects in the research group obviously improved after 3 months of t'ai chi exercise (p < 0.05)., Conclusions: t'ai chi exercise can reduce the level of blood pressure in young and middle-aged in-service staff with grade 1 hypertension, control weight, slow down the HR, improve metabolism, and improve quality of life. t'ai chi is an exercise suitable for in-service hypertension subjects.

Background: Mucin 13 (MUC13) is a cell surface glycoprotein aberrantly expressed in a variety of epithelial carcinomas. Thus far, the role of MUC13 in various diseases remains elusive. To the best of our knowledge, this is the first study to examine the potential of MUC13 as a serum biomarker in a variety of carcinomas and other conditions., Methods: We developed a recombinant MUC13 protein, mouse monoclonal antibodies and enzyme immunoassay (ELISA) for MUC13. We used this assay to measure MUC13 levels in the supernatants of cancer cell lines and a large cohort of serum samples from healthy and diseased individuals., Results: MUC13 is secreted from cancer cell lines, with highest levels found in ovarian cancer cell lines. MUC13 levels in human sera were significantly increased in patients with renal failure and 20%-30% of patients with ovarian, liver, lung and other cancers. MUC13 was also elevated in 70% of patients with active cutaneous melanoma, but not uveal melanoma. Furthermore, we identified significant MUC13 elevations in the serum of patients with vasculitis (ANCA-positive) autoantibodies, but not in those with inflammatory bowel disease., Conclusions: Serum MUC13 is frequently elevated not only in a variety of malignant cases but also in some benign pathologies, thus appearing to be a non-specific disease biomarker. Nonetheless, serum MUC13 is clearly highly elevated in some carcinoma patients, and its relationship with tumor progression in this context warrant further research. Future studies that examine the correlation between serum MUC13 levels to stage of cancer could elucidate prognostic potential.

Background: Human tissue kallikrein 15 (KLK15) is the latest member of the kallikrein-related peptidase family. Little is known about the pathophysiological roles of KLK15. Previous studies implied a role of KLK15 in prostate cancer., Methods: In the present study, we examined KLK15 protein expression using a new immunoassay (ELISA) and immunohistochemistry (IHC)., Results: Highest KLK15 levels were detected in the testis and seminal fluid, whereas lower levels were observed in prostate and other tissues. Immunohistochemical analysis of testis suggests that KLK15 is strongly expressed in mature spermatids, but not in immature germ cells. KLK15 displayed predominantly nuclear localization in the basal cell layer of the prostatic epithelium. We also measured KLK15 in supernatants of various cell lines. Highest KLK15 levels were primarily detected in prostate cancer cell lines and KLK15 expression was hormone-independent, in contrast to KLK3., Conclusions: Collectively, our data provide insights into the localization and possible role of KLK15 in human physiology., (Copyright © 2018 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.)

Objective: Human tissue kallikrein 15 (KLK15) is the last cloned member of the KLK-related gene family. Despite being implicated in multiple cancers, its pathophysiological role remains unknown. We aimed to biochemically characterize KLK15 and preliminarily study its role in cancer., Design & Methods: Recombinant KLK15 protein was produced, purified to homogeneity and quantified by mass spectrometry (parallel reaction monitoring analysis). We profiled the enzymatic activity of KLK15 using fluorogenic peptide substrates, and performed kinetic analysis to discover the cleavage sites. As KLK15 has mainly been associated with prostate cancer, we used a degradomic approach and subsequent KEGG pathway analysis to identify a number of putative protein substrates in the KLK15-treated prostate cancer cell line PC3., Results: We discovered trypsin-like activity in KLK15, finding that it cleaves preferentially after arginine (R). The enzymatic activity of KLK15 was regulated by different factors such as pH, cations and serine protease inhibitors. Notably, we revealed that KLK15 most likely interacts with the extracellular matrix (ECM) receptor group., Conclusion: To our knowledge, this is the first study that experimentally verifies the trypsin-like activity of KLK15. We show here for the first time that KLK15 may be able to cleave many ECM components, similar to several members of the KLK family. Thus the protease could potentially be linked to tumorigenesis by promoting metastasis via this mechanism., (Copyright © 2018 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.)

Objective: To evaluate the effect and safety of Kuanxiong Aerosol (, KA) on patients with angina pectoris., Methods: Block randomization was performed to randomly allocate 750 patients into KA (376 cases) and control groups (374 cases). During an angina attack, the KA group received 3 consecutive sublingual sprays of KA (0.6 mL per spray). The control group received 1 sublingual nitroglycerin tablet (NT, 0.5 mg/tablet). Log-rank tests and Kaplan-Meier estimations were used to estimate the angina remission rates at 6 time-points after treatment (1, 2, 3, 4, 5, and >5 min). Logistic regression analysis was performed to observe the factors inflfluencing the rate of effective angina remission, and the remission rates and incidences of adverse reactions were compared for different Canadian Cardiovascular Society (CCS) classes of angina., Results: The 5-min remission rates in the KA and control groups were not signifificantly different (94.41% vs. 90.64%, P>0.05). The angina CCS class signifificantly inflfluenced the rate of remission (95% confidence interval = 0.483-0.740, P<0.01). In the CCS subgroup analysis, the 3-and 5-min remission rates for KA and NT were similar in the CCSII and III subgroups (P>0.05), while they were signifificantly better for KA in the CCSI and II subgroups (P<0.05 or P<0.01). Furthermore, the incidence of adverse reactions was signifificantly lower in the KA group than in the control group for the CCSII and III subgroups (9.29% vs. 26.22%, 10.13% vs. 20.88%, P<0.05 or P<0.01)., Conclusions: KA is not inferior to NT in the remission of angina. Furthermore, in CCSII and III patients, KA is superior to NT, with a lower incidence of adverse reactions. (Registration No. ChiCTRIPR-15007204).

Objective: To investigate the activity of pyramidal neurons in the medial prefrontal cortex (mPFC) of normal and 6-OHDA-lesioned rats and the responses of the neurons to 5-hydroxytryptamine-7 (5-HT(7)) receptor stimulation., Methods: The changes in spontaneous firing of the pyramidal neurons in the mPFC in response to 5-HT(7) receptor stimulation were observed by extracellular recording in normal and 6-OHDA-lesioned rats., Results: Both systemic and local administration of 5-HT(7) receptor agonist AS 19 resulted in 3 response patterns (excitation, inhibition and no change) of the pyramidal neurons in the mPFC of normal and 6-OHDA-lesioned rats. In normal rats, the predominant response of the pyramidal neurons to AS 19 stimulation was excitatory, and the inhibitory effect of systemically administered AS 19 was reversed by GABAA receptor antagonist picrotoxinin. In the lesioned rats, systemic administration of AS 19 also increased the mean firing rate of the pyramidal neurons, but the cumulative dose for producing excitation was higher than that in normal rats. Systemic administration of AS 19 produced an inhibitory effect in the lesioned rats, which was partially reversed by picrotoxinin. Local administration of AS 19 at the same dose did not change the ?ring rate of the neurons in the lesioned rats., Conclusion: The activity of mPFC pyramidal neurons is directly or indirectly regulated by 5-HT7 receptor, and degeneration of the nigrostriatal pathway leads to decreased response of these neurons to AS 19.

Objective: To evaluate the value of hip MR for diagnosing acetabular labrum tears, and to further compare the diagnostic performances of conventional MR with MR arthrography in acetabular labrum tears., Methods: 90 patients undergoing both hip MR examination and subsequent hip arthroscopy were retrospectively evaluated. Of these patients, 34 accepted both conventional MR and MR arthrography; while the other 56 only underwent conventional MR examination. All hip MR images were independently reviewed by two radiologists, and further compared with the results of hip arthroscopy., Results: 59 of 90 patients were confirmed with acetabular labral tears by hip arthroscopy and 31 without tears. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of conventional MR for evaluating the acetabular labral tears were 61.0%, 77.4%, 83.7% and 51.1% (radiologist A), and 66.1%, 74.2%, 82.9% and 53.4% (radiologist B), respectively, with good consistency between the two observers (K=0.645). The sensitivity, specificity, PPV and NPV of MR arthrography for assessing the acetabular labral tears were 90.5%, 84.6%, 90.5% and 84.6% (radiologist A), and 95.2%, 84.6%, 90.9% and 91.7% (radiologist B), respectively, with excellent good consistency between the two observers (K=0.810). The sensitivity and NPV of MR arthrography for diagnosing the acetabular labral tears were significantly higher than those of conventional MR (both P<0.05)., Conclusion: Hip MR arthrography is a reliable evaluation modality for diagnosing the acetabular labral tears, and its diagnostic performance is superior to that of conventional MR at 3.0 T., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Objective: To evaluate the anginal attack-relieving efficacy and safety of Kuanxiong Aerosol (KA) in patients with coronary heart disease (CHD)., Methods: A total of 780 patients confirmatively diagnosed as CHD angina from November 2011 to December 2012 in 13 medical centers in the mainland area were assigned to 2 groups by blocked randomization, the treatment group (376 cases) and the control group (374 cases). When the angina attacked, patients in the treatment group received sublingual spray three times, 0.6 mL each time, while those in the control group sublingually dissolved Nitroglycerin Tablet (NT), 0.5 mg each tablet. The effective rate of angina relief, efficacy of electrocardiogram (ECG), and the incidence of adverse reactions were observed., Results: The 3 min and 5 min remission rates of angina attack were 53.72% (202/376) and 94.41% (355/376) in the treatment group, and 47.86% (179/374) and 90.64% (339/374) in the control group. The 95% confidence interval (CI) of the difference between the 2 groups of 3 min and 5 min remission rates of angina attacks were [(-1.84%, 12.32%) and (-1.33%, 6.85%) respectively, P > 0.05]. The total improvement rates of ST-T changes in the treatment group and the control group after treatment were 74.07% and 73.13% respectively (P > 0.05). The adverse reaction rate was 9.31 (35/376 cases) in the treatment group and 22.46% (84/374 cases) in the control group (P < 0.01)., Conclusion: KA was not inferior to NT in relieving anginal attacks and improving ischemic ECG changes, and had obviously less adverse reaction.

Purpose: The purpose of this study is to compare MR arthrography in abduction and external rotation (ABER) position with conventional MR arthrography in neutral position for the detection and further classification of anteroinferior labroligamentous lesions., Materials and Methods: Two hundred and twenty-nine cases of shoulder MR arthrography with subsequent arthroscopy were retrospectively evaluated. All MR arthrograms in ABER position and neutral position were independently assessed by two radiologists who were blinded to the arthroscopic findings. Sensitivities and specificities of both positions for detection and further categorization of anteroinferior labroligamentous lesions were calculated and compared using paired McNemar test. K values were calculated to quantify the level of interobserver agreement., Results: At arthroscopy, 24 Bankart lesions, 59 ALPSA lesions, 39 Perthes lesions, 8 GLAD lesions, 4 ALIPSA lesions, 31 nonclassifiable lesions and 60 intact anteroinferior complexes were found. The sensitivity of MR arthrography in ABER position for detecting anteroinferior labroligamentous lesions was significantly higher than that of in neutral position (92.7-94.5% versus 81.8-83%, P<0.05). For the detection rate of the mentioned 6 subtypes of lesions, only the Perthes lesions had significant improvement in ABER position when compared with conventional MR arthrography (observer 1, 61.5-87.2%, P=0.006; observer 2, 69.2-92.3%, P=0.004). MR arthrography in ABER position was more effective in identifying of Perthes lesions (66.7-74.4% versus 35.9-40%, P<0.05) while the conventional MR arthrography was more effective in accurate diagnose of ALPSA lesions (74.6-78.0% versus 54.2-55.9%, P<0.05). No statistically significant difference of the diagnostic accuracy was found between the two positions for Bankart lesions, GLAD lesions, and ALIPSA lesions., Conclusion: MR arthrography in ABER position has more added value on detection of Perthes lesions in evaluation of anteroinferior labroligamentous complex tear., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Objective: To provide the basis for clinical acquired immunodeficiency syndrome (AIDS) surveillance and to avoid cross infection in hospital, we study the infection status of AIDS in Shandong province., Methods: The fourth-generated Akzo's ELISA kit and the fourth generated Immunoluminometric detection reagent were used for HIV antibody screening for 399 303 cases of both inpatients and outpatients from Jan. 2003 to Dec. 2011. Beijing WanTai ELISA kit and Se-marked rapid detection reagent were used for re-detection, and the positive samples were sent to the local CDC for confirming test by Western Blot., Results: The HIV-1 antibody detection results of 129 (0. 3230 per thousand) patients were confirmed to be positive, including 54 (0. 1352 per thousand) cases of outpatients and 75 (0. 1878 per thousand) cases of inpatients. HIV infection rates in outpatients from 2003 to 2011 were 0.050 per thousand, 0.030 per thousand, 0.111 per thousand, 0.120 per thousand, 0.124 per thousand, 0.113 per thousand, 0.148 per thousand, 0.201 per thousand, 0.2152 per thousand; and that in inpatients were 0. 150 per thousand, 0.089 per thousand, 0.138 per thousand, 0. 144 per thousand, 0. 104 per thousand, 0. 132 per thousand, 0. 197 per thousand, 0. 329 per thousand, 0. 313 per thousand respectively. Among these inpatients, there were 61 cases of medical patients and 14 cases of surgical patients, and most were youths and farmers., Conclusions: HIV infection rate was increasing year by year. Most inpatients whose HIV-1 antibody was positive were in the phase of AIDS. Therefore, it's very necessary to execute routine testing for inpatients and outpatients who need special examination for early diagnosis of HIV infection

Objectives: To clarify the morphological features of normal meniscal roots on magnetic resonance (MR) imaging with an isotropic 3D proton density-weighted (PDW) sequence., Materials: 3D PDW MR was performed in sixty-two patients at 3T before knee arthroscopy. MR images of 34 normal medial menisci and 33 intact lateral menisci confirmed by arthroscopy were retrospectively evaluated. MR signals, insertion sites, dimensions and courses of four meniscal roots were recorded., Results: The anterior root of medial meniscus (ARMM) was typically hypointense, while the posterior root of medial meniscus (PRMM) and the anterior root of lateral meniscus (ARLM) were prone to be hyperintense or showing a comblike signal, and the posterior root of lateral meniscus (PRLM) was usually hypointense or comblike on PDW MR images. ARMM and PRLM had more complex and diverse insertion patterns than ARLM and PRMM. There were significant statistical differences of the lengths, widths, heights and course angles among four meniscal roots (all P<0.001)., Conclusions: The signal intensity of each meniscal root can be hypointense, hyperintense, or comblike on 3D PDW MR images. ARMM and PRLM have more complex and diverse insertion patterns than ARLM and PRMM. The dimensions and courses of four meniscal roots all differ., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Hidden Markov Models (HMMs) have been shown to be useful in handwritten pattern recognition. However, owing to their fundamental structure, they have little resistance to unexpected noise among observation sequences. In other words, unexpected noise in a sequence might "break" the normal transmission of states for this sequence, making it unrecognizable to trained models. To resolve this problem, we propose a leave-one-out-training strategy, which will make the models more robust. We also propose a leave-one-out-testing method, which will compensate for some of the negative effects of this noise. The latter is actually an example of a system with a single classifier and multiple classifications. Compared with the 98.00 percent accuracy of the benchmark HMMs, the new system achieves a 98.88 percent accuracy rate on handwritten digits.

Molecular systematics involves the description of the regulatory networks formed by the interconnections between active transcription factors and their target expressed genes. Here, we have determined the activities of 200 different transcription factors in six mouse tissues using an advanced mouse oligonucleotide array-based transcription factor assay (MOUSE OATFA). The transcription factor signatures from MOUSE OATFA were combined with public mRNA expression profiles to construct experimental transcriptional regulatory networks in each tissue. SRF-centered regulatory networks constructed for lung and skeletal muscle with OATFA data were confirmed by ChIP assays, and revealed examples of novel networks of expressed genes coregulated by sets of transcription factors. The combination of MOUSE OATFA with bioinformatics analysis of expressed genes provides a new paradigm for the comprehensive prediction of the transcriptional systems and their regulatory pathways in mouse.

Hematopoiesis undergoes several migrations from yolk sac to liver and spleen, and finally bone marrow until the end of life. A number of investigations have demonstrated that the hematopoietic microenvironment plays very important role in this process. However, the exact mechanisms remain unknown. In order to systematically analyze and understand the role of hematopoietic microenvironment in the regulation and control of hematopoiesis, a microarray containing 588 complementary DNAs was used to compare the gene expressions between those in murine fetal liver and bone marrow cells. The results obtained from array hybridization were analyzed and reconfirmed by using bioinformatics and RT-PCR as well as Northern blot. The results showed that 65 and 131 genes were relatively high expressed in bone marrow and fetal liver cells respectively among 588 known genes in the array-membrane. According to the survey in the PubMed, 39 out of bone-marrow-expressed genes and 71 in fetal-liver-expressed genes were closely related to the hematopoiesis. Further reconfirmation by RT-PCR or Northern blot has demonstrated that CD18, CD44 an d PSGL-1 genes chosen for analysis were highly expressed in adult bone marrow, but unexpressed or lower expressed in fetal liver cells, resulting in high similarity to the array results. Moreover, the expressions of CD18 and CD44 in fetal liver were down-regulated with the increment of gestational age. In conclusion, the gene expressions in bone marrow and fetal liver cells are obviously different, some of the genes are down-regulated at the different stages of ontogeny. The different gene expression levels between bone marrow and fetal liver, especially those genes closely related to the hematopoiesis, may be the molecular basis for the explanation of why hematopoietic stem cells derived from different tissues have different characterizations as well as the differences from the beginning and terminating of fetal liver hematopoiesis, and why hematopoietic stem cells derived from fetal liver is tremendously difficult to be grafted in bone marrow.

Hematopoietic stromal cells, being the essential ingredient of the hematopoietic microenvironment, play very important roles in the control and regulation of self-renewal, proliferation and differentiation of hematopoietic stem cells (HSC) via complex interactions of cell-cell, cell-humoral and cell-extracellular matrix. Evidence from in vivo experiment has proved that HSC derived from normal mice could reconstitute hematopoiesis of mice with HSC defects but failed to reconstitute hematopoiesis of those mice with microenvironment defects, showing the importance of hematopoietic microenvironment in the maintenance of hematopoiesis in vivo. A well-known long-term culture (LTC) system established by Dexter demonstrated in another way that stromal cell layer in the system could support ex vivo hematopoiesis for several months, even more than one year under the optimal conditions. It, however, has not been demonstrated that what is the key elements and in which way the ex vivo hematopoiesis could be maintained for so long time. As the inventions for the large-scale screening methodologies the suppression subtractive hybridization (SSH) was chosen for the screening differentially expressed genes expressed by LTC cultured stromal cells but not by the uncultured bone marrow cells (BMC). mRNA extracted from both cultured adherent cells (tester) and BMC (driver) were hybridized according to the protocol provided by CLONTECH. Total of 130 clones differentially expressed by cultured cells were randomly picked up and 106 ESTs were obtained after sequencing. They represent 26 identical or similar genes and 7 novel genes after the bioinformatics analysis. 5 of the novel genes with the entire open reading frame, without functional clues, have been cloned into the mammalian expression vectors and the functions of them in the control of proliferation and differentiation of HSC will be further exploring. The most interesting discovery is that 3 novel genes have signal peptides, implying the potential discovery of novel growth factors as 80% known growth factors have signal peptides. Our experimental results suggest that: (a) based on the results of subtractive efficiency, the SSH could be a reliable method to screen differentially expressed genes; (b) gene expression may be regulated by multiple factors, even conditioning-dependent, in this experiment the genes expressed by bone marrow stromal cells are LTC-cultivation inducible; (c) it is possible to find interesting genes or special gene after relatively large-scale screen.

The body surface area of 100 Chinese adults (50 for each sex) was measured with the paper cast method and compared to the results estimated by the Stevenson's "height-weight-surface" formula. Our results shows that Stevenson's formula is no longer suitable for the Chinese. Our formulae for Chinese are as follows: SI=0.0061 x H + 0.0124 x W - 0.0099 (for both sexes), SII=0.0057 x H + 0.0121 - W + 0.0882 (for male), SIII=0.0073 H + 0.0127 x W - 0.2106 (for female) H: body height, W: body weight, S: body surface area