Establishment and Evaluation of Intestinal Injury Model of Mouse Acute Graft Versus Host Disease Based on An Organoid Technology / 中国实验血液学杂志

OBJECTIVE@#To establish an intestinal organoid model that mimic acute graft versus host disease (aGVHD) caused intestinal injuries by using aGVHD murine model serum and organoid culture system, and explore the changes of aGVHD intestine in vitro by advantage of organoid technology.@*METHODS@#20-22 g female C57BL/6 mice and 20-22 g female BALB/c mice were used as donors and recipients for bone marrow transplantation, respectively. Within 4-6 h after receiving a lethal dose (8.0 Gy) of γ ray total body irradiation, a total of 0.25 ml of murine derived bone marrow cells (1×107/mice, n=20) and spleen nucleated cells (5×106/mice, n=20) was infused to establish a mouse model of aGVHD (n=20). The aGVHD mice were anesthetized at the 7th day after transplantation, and the veinal blood was harvested by removing the eyeballs, and the serum was collected by centrifugation. The small intestinal crypts of healthy C57BL/6 mice were harvested and cultivated in 3D culture system that maintaining the growth and proliferation of intestinal stem cells in vitro. In our experiment, 5%, 10%, 20% proportions of aGVHD serum were respectively added into the organoid culture system for 3 days. The formation of small intestinal organoids were observed under an inverted microscope and the morphological characteristics of intestinal organoids in each groups were analyzed. For further evaluation, the aGVHD intestinal organoids were harvested and their pathological changes were observed. Combined with HE staining, intestinal organ morphology evaluation was performed. Combined with Alcian Blue staining, the secretion function of aGVHD intestinal organoids was observed. The distribution and changes of Lgr5+ and Clu+ intestinal stem cells in intestinal organoids were analyzed under the conditions of 5%, 10% and 20% serum concentrations by immunohistochemical stainings.@*RESULTS@#The results of HE staining showed that the integrity of intestinal organoids in the 5% concentration serum group was better than that in the 10% and 20% groups. The 5% concentration serum group showed the highest number of organoids, the highest germination rate and the lowest pathological score among experimental groups, while the 20% group exhibited severe morphological destruction and almost no germination was observed, and the pathological score was the highest among all groups(t=3.668, 4.334,5.309,P<0.05). The results of Alican blue staining showed that the secretion function of intestinal organoids in serum culture of aGVHD in the 20% group was weaker than that of the 5% group and 10% of the organoids, and there was almost no goblet cells, and mucus was stainned in the 20% aGVHD serum group. The immunohistochemical results showed that the number of Lgr5+ cells of intestinal organoids in the 5% group was more than that of the intestinal organoids in the 10% aGVHD serum group and 20% aGVHD serum group. Almost no Clu+ cells were observed in the 5% group. The Lgr5+ cells in the 20% group were seriously injuried and can not be observed. The proportion of Clu+ cells in the 20% group significantly increased.@*CONCLUSION@#The concentration of aGVHD serum in the culture system can affect the number and secretion function of intestinal organoids as well as the number of intestinal stem cells in organoids. The higher the serum concentration, the greater the risk of organoid injury, which reveal the characteristics of the formation and functional change of aGVHD intestinal organoids, and provide a novel tool for the study of intestinal injury in aGVHD.

Research progress on traditional Chinese medicine in treatment of neurodegenerative diseases by delaying neurovascular unit aging / 中国中药杂志

Neurodegenerative diseases are a collective term for a large group of diseases caused by degenerative changes in nerve cells. Aging is the main risk factor for neurodegenerative diseases. The neurovascular unit(NVU) is the smallest functional unit of the brain, which regulates brain blood flow and maintains brain homeostasis. Accelerated aging of NVU cells directly impairs NVU function and leads to the occurrence of various neurodegenerative diseases. The intrinsic mechanisms of NVU cell aging are complex and involve oxidative stress damage, loss of protein homeostasis, DNA damage, mitochondrial dysfunction, immune inflammatory response, and impaired cellular autophagy. In recent years, studies have found that traditional Chinese medicine(TCM) can inhibit NVU aging through multiple pathways and targets, exerting a brain-protective effect. Therefore, this article aimed to provide a theoretical basis for further research on TCM inhibition of NVU cell aging and references for new drug development and clinical applications by reviewing its mechanisms of anti-aging, such as regulating relevant proteins, improving mitochondrial dysfunction, reducing DNA damage, lowering inflammatory response, antioxidant stress, and modulating cellular autophagy.

Expression and biological role of DCST1-AS1 in non-small cell lung cancer / 局解手术学杂志

Objective To detect the expression of DCST1-AS1 in non-small cell lung cancer and explore its effect on malignant biological behavior of cancer cells.Methods The tissues and corresponding adjacent tissues from 65 patients with non-small cell lung cancer were collected,and the normal human bronchial epithelial cells 16HBE and the non-small cell lung cancer cell lines(A549,H1299,H1650 and HCC827)were cultured in vitro.The expression levels of DCST1-AS1 and miR-29b in tissues and cells were detected by RT-qPCR assay,and the correlation between the DCST1-AS1 expression and the clinical characteristics of patients with non-small cell lung cancer were analyzed.A549 cells were divided into the control group,the si-NC group,the si-DCST1-AS1 group,the si-DCST1-AS1+ anti-NC group and the si-DCST1-AS1+anti-miR-29b group.The cell proliferation was detected by CCK-8 assay and clone formation assay,the invasion and migration of cells were detected by Transwell;the expression of E-cadherin,Vimentin and N-cadherin was detected by Western blot.Results The expression of DCST1-AS1 increased and the expression of miR-29b decreased in non-small cell lung cancer tissues and cells(P<0.05).The expression of DCST1-AS1 was correlated with TNM stage,differentiated degree of tissue,lymph node metastasis and pathological types in non-small cell lung cancer patients(P<0.05).Compared with the control group or the si-NC group,the expression of DCST1-AS1,OD value,number of colony-forming cells,migration cells and invasion cells and the expression of Vimentin and N-cadherin in A549 cells of the si-DCST1-AS1 group decreased(P<0.05),while the expression of miR-29b and E-cadherin increased(P<0.05).Knocking down miR-29b could significantly reduce the effect of down-regulation of DCST1-AS1 expression on the malignant biological behavior of A549 cells.Conclusion DCST1-AS1 is highly expressed in non-small cell lung cancer,knocking down DCST1-AS1 may inhibit the malignant biological behavior of non-small cell lung cancer cells by up-regulating the expression of miR-29b.

A Propensity Score Matching Study of Autologous Hematopoietic Stem Cell Transplantation and New Drug Chemotherapy for Newly Diagnosed Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To compare the clinical efficacy, survival, and prognosis of autologous hematopoietic stem cell transplantation (ASCT) with new drug chemotherapy in the treatment of newly diagnosed multiple myeloma (NDMM) in the new drug era.@*METHODS@#The clinical data of 149 patients with NDMM treated with new drug induction regimen in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 2012 to December 2019 were retrospectively analyzed. Twenty-four patients who received ASCT were in ASCT group, and 125 patients who did not receive ASCT were in non-ASCT group. The median follow-up time was 43 (1-90) months. The propensity score matching (PSM) method was used to balance confounding factors, then depth of response, overall survival (OS), and progression-free survival (PFS) between the two groups were compared and subgroup analysis was performed.@*RESULTS@#After matching, the covariates were balanced between the two groups. Fifty-one patients (15 cases in ASCT group and 36 cases in non-ASCT group) were included. ASCT patients had a better complete response (CR) rate than non-ASCT patients receiving maintenance therapy (93.3% vs 42.3%, P=0.004), while there were no statistical differences in deep response rate and overall response rate (ORR) between the two groups (93.3% vs 65.4%, P=0.103; 93.3% vs 96.2%, P=1.000). Before matching, the 3 and 5-year PFS rate and median PFS (mPFS) in ASCT group and non-ASCT group were [89.6% vs 66.5%, P=0.024; 69.8% vs 42.7%; non-response (NR) vs 51.0 months], and the 3 and 5-year OS rate and median OS (mOS) were (100% vs 70.6%, P=0.002; 92.3% vs 49.6%; NR vs 54.0 months). After matching, the 3 and 5-year PFS rate and mPFS in ASCT group and non-ASCT group were (83.6% vs 61.7%, P=0.182; 62.7% vs 45.7%; NR vs 51.0 months), the 3 and 5-year OS rate and mOS were (100% vs 65.6%, P=0.018; 88.9% vs 46.9%; NR vs 51.0 months). Subgroup analysis showed that patients with mSMART 3.0 high risk stratification, the 3-year PFS rate and mPFS in ASCT group and non-ASCT group were (83.3% vs 41.5%, P=0.091; NR vs 34.0 months), and the 3-year OS rate and mOS were (100% vs 41.5%, P=0.034; NR vs 34.0 months). Patients with mSMART 3.0 standard risk stratification, the 3-year PFS rate and OS rate in ASCT group and non-ASCT group were (83.3% vs 76.8%, P=0.672; 100% vs 87.2%, P=0.155). The 3-year PFS and OS rate in MM patients who achieved deep response within 3 months after transplantation compared with non-ASCT patients who achieved deep response after receiving maintenance therapy were (83.1% vs 56.7%, P=0.323; 100% vs 60.5%, P=0.042), and the 3-year PFS and OS rate in patients who achieved overall response in both groups were (83.1% vs 62.5%, P=0.433; 100% vs 68.1%, P=0.082). After matching, Cox multivariate regression analysis showed that mSMART 3.0 risk stratification and ASCT were independent prognostic factors for OS.@*CONCLUSION@#In the new drug era, ASCT can increase CR rate and prolong OS of NDMM patients. ASCT patients who are mSMART 3.0 high risk stratification or achieved deep response within 3 months after transplantation have better OS than non-ASCT patients receiving new drug chemotherapy. ASCT and mSMART 3.0 risk stratification are independent prognostic factors for OS in NDMM patients.

Discovery and confirmation of protein action site AK1 of ginsenosides in brain based on DARTS technology / 中国中药杂志

This study aims to explore the targets of ginsenosides in brain based on drug affinity responsive target stability(DARTS) technology. Specifically, DARTS technology was combined with label-free liquid chromatography tandem mass spectrometry(LC-MS) to screen out the proteins in the brain that might interact with ginsenosides. Based on the screening results, adenylate kinase 1(AK1) was selected for further confirmation. First, the His-AK1 fusion protein was yielded successively through the construction of recombinant prokaryotic expression vector, expression of target protein, and purification of the fusion protein. Biolayer interferometry(BLI) was employed to detect the direct interaction of Rg_1, Re, Rb_1, Rd, Rh_2, F1, Rh_1, compound K(CK), 25-OH-PPD, protopanaxa-diol(PPD), and protopanaxatriol(PPT) with AK1, thereby screening the ginsenoside monomer or sapogenin that had strong direct interaction with the suspected target protein AK1. Then, the BLI was used to further determine the kinetic parameters for the binding of PPD(strongest interaction with AK1) to His-AK1 fusion protein. Finally, molecular docking technology was applied to analyze the binding properties between the two. With DARTS and LC-MS, multiple differential proteins were screened out, and AK1 was selected based on previous research for target verification. Fusion protein His-AK1 was obtained by prokaryotic expression, and the response(nm) of Re, Rg_1, Rd, Rb_1, Rh_1, Rh_2, F1, PPT, PPD, 25-OH-PPD, and CK with His-AK1 was respectively 0.003 1, 0.001 9, 0.042 8, 0.022 2, 0.013 4, 0.037 3, 0.013 9, 0.030 7, 0.140 2, 0.016 0, and 0.040 8. The K_(on), K_(off), and K_D values of PPD and His-AK1 were determined by the BLI as 1.22×10~2 mol~(-1)·L·s~(-1), 1.04×10~(-2) s~(-1), 8.52×10~(-5) mol·L~(-1). According to the molecular docking result, PPD bound to AK1 with the absolute value of the docking score of 3.438, and hydrogen bonds mainly formed between the two. Thus, AK1 is one of the protein action sites of ginsenosides in the brain. The direct interaction between ginsenoside metabolite PPD and AK1 is the strongest.

Potential Biomolecules Capable of Assessing Risk of Osteoporotic Fracture: A Scoping Review / 中国医学科学杂志(英文版)

Osteoporosis as a systemic chronic skeletal disease is characterized by low bone mineral density and increased risk to osteoporotic fractures. Osteoporosis is prevalent in the middle-aged and elderly population, especially in the postmenopausal women. With population aging, osteoporosis has become a world-wide serious public health problem. Early recognition of the high-risk population followed by timely and efficient intervention and/or treatment is important for preventing osteoporotic fractures. In light of the high heritability and complex pathogenesis of osteoporosis, comprehensive consideration of vital biological/biochemical factors is necessary for accurate risk evaluation of fractures. For this purpose, we review recent research progress on molecules which can be applied to assess risk for osteoporotic fractures. Future integrative analyses and systematic evaluation of these molecules may facilitate developing novel methodologies and/or test strategies, i.e., biochips, for early recognition of osteoporosis, hence contributing to preventing osteoporotic fractures.

Effect of small ubiquitin-like modifier 1 pseudogene 3 on the proliferation and apoptosis of non-small cell lung cancer cell line H1299 through protein kinase B signaling pathway / 解剖学报

Objective To investigate the effect of long noncoding RNA (IncRNA) small ubiquitin-like modifier 1 pseudogene 3(SUM01P3) on the proliferation and apoptosis of non-small cell lung cancer cell line 1299. Methods Determination of SUM01P3 expression in non-small cell lung cancer cells by Real-time PCR. SUM01P3 small interfering RNA(siRNA) was transfected into H1299 cells, the down regulation effect was determined by Real-time PCR. Cell proliferation was measured by MTT, 5-ethynyl-2′-deoxyuridine(EdU) method, the cell cycle was determined by PI single staining, apoptosis was detected by annexin V -FITC/PI, detection of apoptosis by TUNEL, Western blotting was used to detect the expression of cleaved Caspase-3 (c-Caspase-3), cyclin Dl, P27, phosphorylated phospoinositide 3-kinase (p-PI3K) and phosphorylated protein kinase B (p-Akt). Akt signal activator treated H1299 cells transfected with SUM01P3 siRNA, cell proliferation, apoptosis and cycle change were also measured by the above methods. The number of samples was 9. Results SUM01P3 was up-regulated in non-small cell lung cancer cells. The expression of SUM01P3 in H1299 cells decreased after transfection with SUM01P3 siRNA, cell proliferation decreased, the ratio of G0/Gj phase increased, apoptosis rate increased, c-Caspase-3 and P27 protein in the cells increased, the protein levels of cyclin D1, p-PI3K and p-Akt decreased. Akt signal activator could reverse the inhibition of proliferation, cycle arrest and apoptosis of H1299 cells by SUM01P3 siRNA. Conclusion Down-regulation of SUM01P3 inhibits the proliferation of non-small cell lung cancer H1299 cells and induces apoptosis, the mechanism of action is related to the reduction of the activation level of the Akt signaling pathway.

Research Progress of Regulatory Effect and Mechanism of Flavonoids on Vascular Endothelial Cell Injury / 中国实验方剂学杂志

To investigate the effect of flavonoid compounds on vascular endothelial cells. Vascular endothelial cells were located between plasma and vascular tissue, and can complete the metabolic exchange of plasma and interstitial fluid, synthesize and secrete a variety of biologically active substances, so as to ensure the normal contraction and relaxation of blood vessels, and maintain the tension of blood vessels. Besides, it can regulate blood pressure and the balance of blood coagulation and anticoagulation, and maintain normal blood flow and long-term patency of blood vessels. Endothelial cell damage can cause a series of cardiovascular diseases, such as hypertension and coronary heart disease. Flavonoids are widely found in nature. Because these compounds are mostly yellow or light yellow, they contain ketone groups in the molecule, which are called flavones. Flavonoids are widely distributed, mostly in higher plants and ferns. Various flavonoid compounds, such as flavonoids, flavonols, flavanones isoflavones and flavanones, can protect vascular endothelial cells. This article reviews relevant findings published in domestic and foreign journals. It is found that flavonoids have effects in resisting inflammation, reducing blood vessel fragility, improving blood vessel permeability, lowering blood lipids and cholesterol, vasodilating and resisting hemagglutinating, with the same effect as phytoestrogens. They can reduce vascular endothelial cell damage through anti-inflammatory, anti-oxidative stress, stable mitochondrial function, and regulating nitric oxide（NO）. It can be used in clinic to treat diseases, such as insufficient cerebral blood supply, sequelae caused by cerebral hemorrhage, hyperviscosity, cerebral thrombosis, coronary heart disease and angina pectoris.

Research progress on anti-atherosclerosis effect and mechanism of flavonoids compounds mediated by macrophages / 中国中药杂志

Flavonoids are important active ingredients of traditional Chinese medicine, mainly with cardiovascular, anti-liver injury, antioxidant, antispasmodic, and estrogen-like effects. These compounds have obvious effects on the cardiovascular and cerebrovascular diseases. Macrophage-derived foam cells are the key medium in the process of atherosclerosis(AS). In plaque, allserum lipids, serum lipoproteins, and various pro-or anti-inflammatory stimulating factors, chemokines, and small bioactive molecules can significantly affect the macrophage phenotype and induce stronger pro-inflammatory or anti-inflammatory properties. Studies have shown that some flavonoids can be used for macrophages through different pathways and mechanisms, playing an anti-atherosclerosis effect to different degrees, including promotion of cholesterol efflux from macrophages, anti-foaming of macrophages, inhibition of secretion of inflammatory factors, and antioxidant modified low density lipoprotein(ox-LDL)-induced apoptosis of macrophages. Related gene regulation inclu-ded ATP-binding cassette transporter A1(ABCA1), ATP-binding cassette transporter G1(ABCG1), Toll-like receptor(TLR), and scavenger receptor(SR). In this article, we would review the recent research progress of flavonoids on anti-atherosclerosis effect me-diated by macrophage. It is expected to provide new treatment strategies for AS-related cardiovascular and cerebrovascular diseases, and provide research ideas and development directions for the use of related natural medicines and design of new products.

Effect of 's subcutaneous needling on thickness and elasticity of affected muscles in shoulder neck pain based on ultrasonic elastography / 中国针灸

OBJECTIVE@#To observe the effect of 's subcutaneous needling on thickness and elasticity of affected muscles in subjects with shoulder neck pain by ultrasonic elastography.@*METHODS@#A total of 30 subjects with upper trapezius pain and local tenderness, or stiffness and funicular nodules were observed randomly, right shoulders were as observation group and left shoulders were as control group. Simple resistance training was adopted in the control group. At the same time of the resistance training, sweeping technique of 's subcutaneous needling was adopted at the local tenderness or the stiffness and funicular nodules of upper trapezius in the observation group. The treatment was given once in both groups. Before and immediately after treatment, thickness and elasticity of bilateral upper trapezius and supraspinatus were observed by ultrasonic elastography, and the variations of visual analogue scale (VAS) score were observed in the two groups.@*RESULTS@#Compared before treatment, the elasticity of upper trapezius and supraspinatus were decreased after treatment in both groups (<0.05), and those in the observation group were lower than the control group (<0.05). Compared before treatment, the VAS scores after treatment were decreased in both groups (<0.05), and that in the observation group was lower than the control group (<0.05).@*CONCLUSION@#'s subcutaneous needling can increase the elasticity and release the muscular tension of affected muscles, and relieve pain in subjects with shoulder neck pain.

Effects of different nitrogen sources on accumulation of active components in hairy roots of Salvia miltiorrhiza and Salvia castanea f. tomentosa / 中草药

Objective: Nitrogen is an important element affecting the accumulation of effective components in Chinese medicinal materials. The purpose of this study was to investigate effects of different nitrogen sources on the growth and active components accumulation of hairy roots of Salvia miltiorrhiza and Salvia castanea f. tomentosa. Methods: The hairy roots of S. miltiorrhiza and S. castanea f. tomentosa were treated with ammonium nitrate, hydrolyzed milk protein, peptone, beef extract, casein and yeast extract, respectively. The growth of hairy roots and the accumulation of active components were analyzed. Results: Ammonium nitrate was the most beneficial to the growth of the two kinds of hairy roots. Hydrolyzed milk protein significantly promoted the accumulation of salvianolic acids, compared with ammonium nitrate, the contents of rosmarinic acid and salvianolic acid B in S. miltiorrhiza were respectively increased by 2.94 times and 3.27 times, and the contents of rosmarinic acid and salvianolic acid B in S. castanea f. tomentosa were respectively increased by 13.74 times and 2.01 times. Yeast extract had the most significant effect on the accumulation of dihydrotanshinone I and cryptotanshinone in two kinds of hairy roots. Hydrolyzed milk protein significantly promoted the accumulation of tanshinone IIA in the hairy roots of S. miltiorrhiza, while beef extract had the most significant effect on the accumulation of tanshinone IIA in S. castanea f. tomentosa. Conclusion: Ammonium nitrate was the best nitrogen source for the growth of two kinds of hairy roots, and hydrolyzed milk protein was the best nitrogen source for salvianolic acids accumulation. The effects of different nitrogen sources on four kinds of tanshinones were different, and the responses of S. miltiorrhiza and S. castanea f. tomentosa to different nitrogen sources were also different. This study not only has certain guiding significance for large-scale cultivation of hairy roots of S. miltiorrhiza and industrialized production of active components, but also provides a reference for the development and utilization of S. castanea f. tomentosa resources.

Effects of different carbon sources on growth and active component contents in Salvia miltiorrhiza and S. castanea f. tomentosa hairy roots / 中国中药杂志

Salvia miltiorrhiza(Sm) and Salvia castanea f. tomentosa(Sc) hairy roots were used as experimental materials to study the effects of six different carbon sources, galactose, fructose, lactose, glucose, arabinose and sucrose(control), on fresh weight, dry weight, contents and yields of salvianolic acids and tanshinones. The results showed that galactose was most beneficial to the growth of two kinds of hairy roots, while lactose and arabinose were not conducive to their growth. As for Sm hairy roots, fructose significantly promoted the accumulation of salvianolic acid B, and the content increased by 5.801 times and 10.151 times compared with the control group, respectively. Glucose significantly promoted the accumulation of salvianolic acids. The content and yield of rosmarinic acid were 7.674 times and 9.260 times of that of the control group, and the content and yield of salvianolic acid B were 5.532 times and 6.675 times of the control group. For the hairy roots of Sc, galactose significantly increased the content and yield of rosmarinic acid, reaching 7.820 times and 9.944 times of the control group, respectively. Fructose promoted the increase of the content and yield of cryptotanshinone, reaching 9.242 times and 6.609 times of the control group, respectively. The study confirmed the optimal carbon source for the hairy root culture of Sm and Sc, and provided theoretical guidance for large-scale production of Sm drug-derived components and the utilization of Sc.

Recent Progress of Molecular Genetic Characteristics and Prognosis of Non-Hodgkin's Lymphoma--Review / 中国实验血液学杂志

Lymphomas are traditionally divided into Hodgkin's lymphoma and non-Hodgkin's lymphoma(NHL), the NHL is a common hematological cancer, which represents a wide spectrum of illnesses from the most indolent to the most aggressive malignancies, and the detection of related molecular targets will be needed for diagosing each subtype of NHL. Advances in understanding the pathogenesis of NHL have improved the precision of diagnosis and the prognosis evaluation of patients with this disorder, such as chromosomal translocation leading to the up-regulation of oncogene expression. Besides, the deletion of several tumor suppressor genes may cause excessive proliferation in tumor cells, and the single nucleotide polymorphism (SNP) determines the differences of susceptibility, drug-resistance and prognosis of NHL. In addition, DNA methylation, histone modification, non-coding RNA and other epigenetic phenomena play an increasingly important role in the diagnosis, selection of clinical drugs and evaluation of prognosis of NHL. In this review, the recent progress of researches on chromosome translocation, deletion of tumor suppression genes, gene poly-morphism and epigenetics are summarized.

Analysis of Clinical Pathological Features and Prognosis in Young Patients with Diffuse Large B Cell Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To explore the clinical pathological features of the patients with diffuse large B cell lymphoma (DLBCL) and their prognostic factors.@*METHODS@#The prognosis of the clinical pathological features and their influence on prognosis of 177 patients diagnosed as DLBCL at the first visit from January 2013 to May 2017 in our hospital were analyzed retrospectively.@*RESULTS@#The univariate analysis showed that overall survival (OS) and progression-free survival (PFS) were associated with later Ann Arbor stage (Ⅲ-Ⅳ) ( P＜0.01, P＜0.05), high performance status (ECOG score 2-4) (P＜0.01, P＜0.05), extranodal involvement ＞1 (P＜0.01, P＜0.05), elevated LDH level (P＜0.01, P＜0.05). B symptom (P＜0.05) and elevated β2-MG level (P＜0.05) also influenced OS. COX multivariate analysis showed that the elevated β2-MG level (P＜0.05) and later stage (Ⅲ-Ⅳ) (P＜0.05) have an independent influence on OS, later stage (Ⅲ-Ⅳ) (P＜0.05) also independently influenced PFS. The patients with high aaIPI score (2-3) and bone marrow involvement before treatment had poor OS (P＜0.01, P＜0.01) and PFS (P＜0.05, P＜0.01).@*CONCLUSION@#Elevated β2-MG level can independently influence OS, and later stage (Ⅲ-Ⅳ) can independently influence both OS and PFS. High aaIPI score (2-3) and bone marrow involvement before treatment have an inferior influence on OS and PFS.

Integrative Analysis Confirmed the Association between Osteoprotegerin and Osteoporosis / 中国医学科学杂志(英文版)

Objective This study aimed to verify the association between osteoprotegerin gene () and its variants with osteoporosis (OP) by performing integrative analysis.Methods We used the KGG software to perform gene-based association analysis, which integrated all publicly available single-nucleotide polymorphism (SNP)-based values and obtained an overall value for the . The significant SNPs were screened for expression quantitative trait loci (eQTLs). Meta-analysis was used to combine the associations between the variants of and bone mineral density (BMD) reported in the literatures. Then we performed dual-luciferase reporter gene systems for the functional verification of the variants of .Results In the gene-based association analysis, the over all value of was 6.24×10 for BMD at femoral neck (FN) and 7.37×10 for BMD at lumbar spine (LS), indicating the importance of for OP. The publicly available eQTL database identified 5 eQTLs which exert cis-regulation effects on at FN and LS. Literature searching found that rs2073617 (known as T950C) was the hot spot SNP. There were 13 relevant studies on rs2073617 besides the GEFOS-2 study identified from the PubMed. Significant differences among TT, TC and CC genotypes at FN (= 0.047) and LS (= 0.025) were shown by meta-analysis, demonstrating the associations between T950C polymorphism and BMD. Luciferase gene expression was significantly higher at the presence of allele C than allele T in the 293T cells (=-9.47, <0.01). Conclusion The integrative analysis further confirmed the importance of in OP and the correlation of T950C polymorphism with BMD of OP. The strategy can be used as a reference for functional interpretation of other disease-related genes.

Establishment of A20 Murine B Lymphoma Transplantation Model Labeled with Luciferase / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To establish the animal model of luciferase-transfected A20 murine B cell lymphoma, so as to provide experimental tools to explore the effect of graft versus tumor.</p><p><b>METHODS</b>Luciferase- labeled A20 cells were cloned with puromycin selection. Transfected A20 cells and CBL/6 bone marrow were inoculated into the irradiated BALB/c mice by injection in tailvein to establish the transplantation model. The bioluminescent imaging technique was used to monitor the tumor growth, and then the survival, body weight, tumor formation and pathological characteristics of target organs were observed.</p><p><b>RESULTS</b>A20 cell line stably expressing luciferase gene was successfully obtained. The the bioluminicent imaging found that the tumor luminescence could be observed on day 8 of A20 cell inoculation, and the mean fluorescent intensity was increased along with the tumor growth. Compared with the BMT group, the survival rate and body weight of BMT+A20-Lucmice were decreased significantly. General anatomy showed the tumor mainly formed in the liver and spleen.</p><p><b>CONCLUSION</b>A mouse transplantation model with luciferase- transfected A20 cells has been successfully established, thus laying a foundation for investigation of graft-versus-tumor.</p>

Application of Chimeric Antigen Receptor-Modified NK Cells in Multiple Myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the killing effect of CAR (CD138-CD28-CD3ζ)-NK cells on myeloma cells through construction of CAR(CD138-CD28-CD3)-NK cells.</p><p><b>METHODS</b>The antiCD138scFv-CD28-CD3 zeta plasmid pcDNA3.1 was constructed, which then together with 3 plasmid lentiviral packaging system were transfected into 293T cells, the virus was collected. Furthermore, in order to get the stably transfected cell line, the NK92MI cell line was infected by the virus, then the positive cells were screened by puromycin. The expression of the CARNK cells were verified by RT-PCR and Western blot. At last the ability of secreting cytokine CD107a was detected by flow cytometry, and the statistical analysis was carried out to verify the anti-myeloma effect of CAR-NK cells.</p><p><b>RESULTS</b>Gene fragment of the CAR(antiCD138scFv-CD28-CD3ζ) was constructed successfully by gene engineering technique in vitro, and the gene sequence was verified to be correct by sequencing. By virus packaging technology, the virus expressing the protein of the CAR was obtained. PCR and Western blot verified the expression of CAR fusion protein on the sufurce of NK cells. The cell killing experiment confirmed that the CAR-NK cells possessed the ability to secrete cytokine CD107a superior to control cells and showed the obvious killing effect on multiple myeloma cells.</p><p><b>CONCLUSION</b>The CAR can be constructed in vitro, and express on NK92 cells. The CAR-NK cells can kill the multiple myeloma cells expressing CD138 antigen, thereby plays an antimyeloma effect.</p>

LIMR is involved in the inhibitory effect of antiflammin-1 on epithelial-mesenchymal transition in A549 cells / 生理学报

Epithelial-mesenchymal transition (EMT) occurring in alveolar epithelial cells plays an important role in the development and progression of pulmonary fibrosis. Previous studies showed that antiflammin-1 (the active fragment of uteroglobin) effectively inhibited bleomycin-induced pulmonary fibrosis. However, its mechanism is still far from being clarified. In this study, we investigated the effects of antiflammin-1 on EMT in A549 cells induced by transforming growth factor-β1 (TGF-β1) and the underlying mechanism by using morphological observation and Western blot. The results showed that the expression of α-smooth muscle actin (α-SMA) increased significantly while the expression of E-cadherin decreased significantly in A549 cells following treatment with TGF-β1 concomitant with morphological change of A549 cells from pebble-like shape epithelial cells to spindle-like mesenchymal shape. This process of EMT in A549 cells induced by TGF-β1 was significantly inhibited when A549 cells were co-incubated with TGF-β1 and antiflammin-1. Furthermore, the anti-lipocalin interacting membrane receptor (LIMR) antibody and PD98059 (an ERK signaling pathway blocker) attenuated the inhibitory effect of antiflammin-1 on TGF-β1-induced EMT, respectively. Our findings indicate that antiflammin-1 can inhibit EMT in A549 cells induced by TGF-β1, which is related to LIMR and its downstream ERK signaling pathway.

Treatment of Chronic Graft versus Host Disease -Review / 中国实验血液学杂志

Chronic graft versus host disease (cGVHD) is the major reason of late non-relapse related death after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The therapeutic regimen based on glucocorticoids remains as the standard initial treatment of cGVHD. But due to the significant side effects and unsatisfactory outcomes, particularly for patients with steroid-refractory disease, the more effective and less toxic therapies should be needed. Abnormal regulation for the stable state of T lymphocytes and B lymphocytes for allo-HSCT patients is the current focus of studying cGVHD pathogenesis. This article summarizes the treatment of cGVHD targeted to pathogenic factors and signaling pathways in pathogenetic process.

Advances of Diagnostic Biomarkers on Acute Graft Versus Host Disease -Review / 中国实验血液学杂志

Acute graft-versus-host disease (aGVHD) is one of the main complications after allogeneic hematopoietic stem cell transplantation (HCT), resulting in considerable morbidity and mortality. Diagnosis of aGVHD is typically based on clinical presentation and confirmed by biopsy. In recent years, aGVHD researches have been focused on diagnostic biomarkers, and several microRNAs, cytokines and immunoregulated cells have been dicovered currently. This review summarizes the advances on aGVHD biomarkers.