Inhibitory Effect of Cinobufotalin on Macrophage Inflammatory Factor Storm and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the inflammatory effects of Cinobufotalin on monocytes in resting state and macrophages in activated state and its molecular mechanism.@*METHODS@#THP-1 cells were stimulated with Phorbol 12-myristate 13-acetate to induce differentiation into macrophages. Lipopolysaccharides was added to activate macrophages in order to establish macrophage activation model. Cinobufotalin was added to the inflammatory cell model for 24 h as a treatment. CCK-8 was used to detect cell proliferation, Annexin V /PI double staining flow cytometry was used to detect cell apoptosis, flow cytometry was used to detect macrophage activation, and cytometric bead array was used to detect cytokines. Transcriptome sequencing was used to explore the gene expression profile regulated by Cinobufotalin. Changes in the significantly regulated molecules were verified by real-time quantitative polymerase chain reaction and Western blot.@*RESULTS@#1∶25 concentration of Cinobufotalin significantly inhibited the proliferation of resting monocytes(P<0.01), and induced apoptosis(P<0.01), especially the activated macrophages(P<0.001, P<0.001). Cinobufotalin significantly inhibited the activation of macrophages, and significantly down-regulated the inflammatory cytokines(IL-6, TNF-α, IL-1β, IL-8) released by activated macrophages(P＜0.001). Its mechanism was achieved by inhibiting TLR4/MYD88/P-IκBa signaling pathway.@*CONCLUSION@#Cinobufotalin can inhibit the inflammatory factors produced by the over-activation of macrophages through TLR4/MYD88/P-IκBa pathway, which is expected to be applied to the treatment and research of diseases related to the over-release of inflammatory factors.

Establishment of a Patient-Derived T-Cell Acute Lymphoblastic Leukemia Xenograft Model in Novel Immunodeficient NCG Mice / 中国实验血液学杂志

OBJECTIVE@#The leukemia cells from patients with T-cell acute lymphoblastic leukemia (T-ALL) were inoculated into NCG mice to establish a stable human T-ALL leukemia animal model.@*METHODS@#Leukemia cells from bone marrow of newly diagnosed T-ALL patients were isolated, and the leukemia cells were inoculated into NCG mice via tail vein. The proportion of hCD45 positive cells in peripheral blood of the mice was detected regularly by flow cytometry, and the infiltration of leukemia cells in bone marrow, liver, spleen and other organs of the mice was detected by pathology and immunohistochemistry. After the first generation mice model was successfully established, the spleen cells from the first generation mice were inoculated into the second generation mice, and after the second generation mice model was successfully established, the spleen cells from the second generation mice were further inoculated into the third generation mice, and the growth of leukemia cells in peripheral blood of the mice in each group was monitored by regular flow cytometry to evaluate the stability of this T-ALL leukemia animal model.@*RESULTS@#On the 10th day after inoculation, hCD45+ leukemia cells could be successfully detected in the peripheral blood of the first generation mice, and the proportion of these cells was gradually increased. On average, the mice appeared listless 6 or 7 weeks after inoculation, and a large number of T lymphocyte leukemia cells were found in the peripheral blood and bone marrow smear of the mice. The spleen of the mice was obviously enlarged, and immunohistochemical examination showed that hCD3+ leukemia cells infiltrated into bone marrow, liver and spleen extensively. The second and third generation mice could stably develop leukemia, and the average survival time was 4-5 weeks.@*CONCLUSION@#Inoculating leukemia cells from bone marrow of patients with T-ALL into NCG mice via tail vein can successfully construct a patient-derived tumor xenografts (PDTX) model.

Infliximab monotherapy for Chinese patients with moderate to severe plaque psoriasis: a randomized, double-blind, placebo-controlled multicenter trial / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Tumor necrosis factor-α is a key mediator in the pathogenesis of psoriasis. Infliximab is a monoclonal antibody that specifically binds to tumor necrosis factor-α. The purpose of this study was to validate the efficacy and safety of 5 mg/kg infliximab therapy in Chinese patients with moderate to severe plaque psoriasis.</p><p><b>METHODS</b>In this multicenter, double-blind, placebo-controlled trial, 129 patients with moderate-to-severe psoriasis were randomized to the induction therapy (weeks 0, 2 and 6) with infliximab 5 mg/kg (n = 84) or placebo (n = 45), followed with infliximab 5 mg/kg scheduled at week 14 and week 22 in the infliximab group, and infliximab 5 mg/kg scheduled at weeks 10, 12 and 16 in the placebo group. The primary end point was the proportion of patients who achieved at least 75% improvement in Psoriasis Area and Severity Index (PASI 75 response rate) from baseline at week 10.</p><p><b>RESULTS</b>At week 10, 81.0% of patients treated with infliximab (5 mg/kg) achieved a 75% or greater improvement compared with 2.2% of patients treated with placebo (P < 0.001). A significant improvement in PASI, Physician's Global Assessment (PGA) and Dermatology Life Quality Index (DLQI), was seen from week 6 through week 14 in the infliximab group compared with the placebo group. Through week 22, PASI, PGA, DLQI were well maintained. The incidence of adverse events for the infliximab treatment group was slightly higher in comparison to the placebo treatment group during the first 10 weeks without statistical significance. However, there were 3 cases of tuberculosis that developed during the 26 weeks treatment with infliximal.</p><p><b>CONCLUSIONS</b>Infliximab treatment was effective as induction and maintenance treatments for Chinese patients with moderate to severe plaque psoriasis. Most drug-induced adverse events were mild to moderate, and well tolerated. Screening for tuberculosis is essential and prophylactic treatment should be given if necessary.</p>

Induction of prostate cancer-specific CTLs with dendritic cells pulsed by different types of tumor antigens / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the effectiveness of freeze-thaw antigens and acid eluted peptide antigens extracted from tumor cell-pulsed dendritic cells (DC) in inducing prostate cancer-specific cytotoxic T lymphocytes (CTL) in vitro.</p><p><b>METHODS</b>Tumor antigens were extracted from the prostate cancer cell line PC-3 with the repeated freeze-thaw and weak acid elution methods. Peripheral blood mononuclear cells were cultured with recombinant human GM-CSF and IL-4 for inducing DCs in vitro. Then the DCs were pulsed with the two kinds of prostate cancer tumor antigens respectively and cultured with T cells for inducing CTLs. The activity of the tumor-specific CTLs were detected by LDH release assay.</p><p><b>RESULTS</b>The protein content in the tumor antigens obtained from PC-3 (2 x 10(7)) by citric acid-phosphate buffer elution and that by the repeated freeze-thaw method were (212.2 +/- 7.9) microg and (963.0 +/- 25.3) microg, respectively. The two kinds of prostate cancer antigens-pulsed DCs had a significant role in inducing the PC-3 cell-specific CTLs, and the CTLs induced by acid-eluted peptide antigen-pulsed DCs exhibited an even more significant tumor-specific cytotoxicity than those induced by repeated freeze-thaw ([60.4 +/- 5.52]% vs. [43.7 +/- 4.11]%, P < 0.01).</p><p><b>CONCLUSION</b>Both the weak acid elution and repeated freeze-thaw methods for extracting prostate cancer antigens can be used for in vitro sensitization of DCs. The DCs pulsed by either of the two kinds of antigens can activate CTLs, and the antigens extracted by weak acid elution are even more effective.</p>

Dendritic cell-based immunotherapy for multiple myeloma -- review / 中国实验血液学杂志

Patients with multiple myeloma (MM) have increased constantly in recent years, but treatment for patients with MM is currently unsatisfactory and it is necessary to develop new complementary therapies. Dendritic cells (DCs) are specialized antigen-presenting cells capable of initiating and regulating immune responses. Vaccination with tumor antigen-pulsed DCs has shown to be safe and possesses therapeutic effect against many tumors. In this review, the various types of MM-associated antigens and clinical trials on DC-based immunotherapy in MM are summarized, the development of DC immunotherapy for MM patients in future trials is discussed.

Surgical treatment and prognosis analysis of localized retroperitoneal Castleman disease: a study of 20 cases / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the clinical characteristics, surgical treatment and prognosis analysis of localized retroperitoneal Castleman disease (CD), and to improve the level of diagnosis and treatment of retroperitoneal Castleman disease with paraneoplastic pemphigus (PNP).</p><p><b>METHODS</b>The clinical data of retroperitoneal CD with PNP from January 1993 to May 2009 were compared with CD without PNP retrospectively, including clinical features, tumor site, lab examination, surgical treatment, pathologic subtype and results of surgery.</p><p><b>RESULTS</b>(1) Retroperitoneal Castleman disease more likely originated in para-kidney and iliac fossa with middle age of 36 years old, especially in left retroperitoneum. Of the 20 cases, 18 tumors (90%) were hyaline vascular variants and 2 were mixed variants. There was no statistical difference in gender, age, tumor site and pathological subtype between two groups. (2) Retroperitoneal CD with PNP more likely complicated with bronchiolitis obliterans (BO) and high level of serum CEA/CA242. (3) Retroperitoneal Castleman tumors had clear margin, intact envelop and were easily resectable, however the biological behavior of CD with PNP might tend malignant changing, invade adjacent blood vessel or seed locally, and eventually relapse after operation. (4) The 5-year survival rate of retroperitoneal CD with PNP was 42.8%, significantly lower than those without PNP. The average survival time was 30 months. Bronchiolitis obliterans and radical resection were the key effect in prognosis of retroperitoneal CD.</p><p><b>CONCLUSIONS</b>Retroperitoneal CD with PNP has distinctive clinical features. Early diagnosis, prompt removal of tumor and termination secretion of causative antibody are critical to the management of this disease.</p>

Basic and clinical research on paraneoplastic pemphigus / 中国医学科学院学报

Paraneoplastic pemphigus (PNP) is an autoimmune blistering skin disease associated with neoplasms. Clinically, it is characterized by severe mucosal erosions and various cutaneous lesions. Suprabasal acantholysis and cleft with scattered necrotic keratinocytes are the unique histopathological features of PNP. The pathogenic autoantibodies existed in PNP sera, and their production was correlated with the associated tumor. Early detection and resection of the tumor are essential for the treatment of the disease.

Role of autoantibodies against the linker subdomains of envoplakin and periplakin in the pathogenesis of paraneoplastic pemphigus / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The presence of autoantibodies against multiple epidermal proteins is an important feature in paraneoplastic pemphigus (PNP). Circulating anti-desmoglein 3 autoantibody, the major pathogenic autoantibody in pemphigus vulgaris (PV), has been proved pathogenic in PNP. Because of many clinical differences between PNP and PV, we speculate about the involvement of other autoantibodies in the pathogenesis of PNP. Envoplakin (EPL) and periplakin (PPL) are recognized by most PNP sera. Their linker subdomains are highly homologous and necessary for the association of intermediate filaments.</p><p><b>METHODS</b>We characterized the autoantibodies against the linker subdomains of EPL and PPL in PNP patients' sera and their associated tumors by enzyme-linked immunosorbent assay (ELISA) and immunofluorence. We also applied the purified autoantibodies against EPL and PPL from PNP sera to cultured human epidermal keratinocytes (HEK), to evaluate the changes of cell-cell adhesion.</p><p><b>RESULTS</b>Autoantibodies against EPL and PPL were detected in most PNP patients by ELISA, and the decrease of these autoantibodies after removal of the tumors was roughly comparable to the improvement of clinical symptoms. Cultured tumor cells from PNP patients secreted these autoantibodies. Specific immunoglobulin receptors for EPL and PPL were found on B lymphocytes in tumors from PNP. Furthermore, purified anti-EPL and anti-PPL autoantibodies from PNP sera were capable of dissociating cultured human epidermal keratinocytes.</p><p><b>CONCLUSION</b>Autoantibodies against EPL and PPL may also be pathogenic in PNP.</p>

Study on the association of clinical characteristic, CD4+ and level of HIV viral load among 690 initial HIV-infection / 中华流行病学杂志

<p><b>OBJECTIVE</b>To understand the correlation between CD4+ cell count, HIV viral load (VL) and clinical characteristics among patients when HIV-1 was tested positive and initial AIDS diagnosis was made.</p><p><b>METHODS</b>690 HIV-infected cases from Beijing Di-Tan Hospital were included and under a cross sectional study while SPSS statistical method was used.</p><p><b>RESULTS</b>The 690 HIV-infected cases would include 458 males and 232 females with age range from 2-72 years (mean age as 35.3). The modes of transmission showed that: homosexual contact taking up 17.5% while heterosexual was 16.7%. Most of the homosexual-infected ones lived in Beijing and most of them had bachelor or master's degrees. 19.4% of the transmission happened between heterosexual/bisexual couples, suggesting that HIV was transmitted through the "bridge population" while the rest were infected by contaminated blood/plasma. Many of the cases were identified when they lately visited the pre-operation surveillance point in the hospital. Serious immunodeficiency symptoms or signs were discovered as: CD4+ count < 50 cell/microl, serious opportunistic infections including pneumocystosis pulmonary, cerebral toxoplasmosis and cryptococcal meningitis. Higher frequencies of diseases seen were dermotosis, pneumonia, upper respiratory tract infection, hepatitis and digestive tract moniliasis.</p><p><b>CONCLUSION</b>Because of the late identification of the disease, serious immuo-suppression situation often appeared, suggesting that there was an urgent need to improve STD/AIDS knowledge on those HIV (+) people so they might have an early access to accept medical care.</p>

Design, synthesis and insulin-sensitizing activity of some peroxisome proliferator-activated gamma agonists / 药学学报

<p><b>AIM</b>To find new peroxisome proliferator-activated y agonists with high activity and low toxicity.</p><p><b>METHODS</b>Based on JTT-501 and JTT-20993, new isoxazolidine-3,5-dione and noncyclic 1,3-dicarbonyl compounds were designed and synthesized. Their insulin-sensitizing activities were evaluated.</p><p><b>RESULTS</b>Eight new compounds were obtained. The structures of synthesized compounds were characterized by NMR, MS and IR. Four compounds (1A-4A) showed insulin-sensitizing activities.</p><p><b>CONCLUSION</b>Compounds (1A and 3A) showed excellent insulin-sensitizing activities and should be worth further investigation.</p>

Cell origin of localized Castleman's disease of hyaline-vascular type / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the cell features of 6 Castleman's disease patients and evaluate their prognosis.</p><p><b>METHODS</b>The resected tumors were investigated by routine histopathology and immunohistochemistry. Reverse-transcript PCR (RT-PCR) and sequencing of RT-PCR products were used to assess the clonal characters of the main tumor cells.</p><p><b>RESULTS</b>Histologically, all of the 6 tumors were classified as the hyaline vascular type. B-cells dominated the follicular germinal centers, with T-cells dispersing inter-follicularly. The results of RT-PCR each obtained a single band of either 128 bp or 122 bp and sequencing showed that there was highly homogeneity within the same length sequences, accompanied by fewer different nucleotide acids.</p><p><b>CONCLUSION</b>Monoclonal and/or oligoclonal B cells were identified in Castleman's disease. These B cells were originated from germinal center cells.</p>

Treatment of paraneoplastic pemphigus with Castleman's disease / 中华外科杂志

<p><b>OBJECTIVE</b>To discuss the clinical findings and treatment of paraneoplastic pemphigus (PNP) with Castleman's disease.</p><p><b>METHODS</b>To investigate the clinical, histopathologic and CT findings of 8 cases paraneoplastic pemphigus with Castleman's disease.</p><p><b>RESULTS</b>All of 8 patients were diagnosed PNP first and were found Castleman's tumor incidently during routine examination. All 8 cases showed severe erosion or ulcer of the oral mucosa with various skin lesions. Histopathologically, there were intraepidermal acantholytic vesicle, basal cell liquefaction, necrotic keratinocytes in the epidermis and lymphocyte infiltration in the upper dermis. CT scan appeared solitary mass in these patients. Some of them were attacked by bronchiolitis obliterans. All 8 patients were failed by use of predisone. Obvious relief of PNP and pulmonary lesion occurred after tumor was rescted.</p><p><b>CONCLUSIONS</b>Paraneoplastic pemphigus with Castleman's disease is a rare disease. The key step is to find and resect the tumor in abdomen. CT scan should be used to detect the tumor in patients with PNP, especially, when predisone was failed in treatment.</p>

IDENTIFICATION OF A NEW TYPE OF AMYLASE AND MUTAGENESIS OF STRAIN ZX99 SECRETING THE ENZYME FOR PRODUCTION OF ISOMALTOOLIGOSACCHARIDE / 微生物学通报

This paper reported a new type of amylase (neoamylase) secreted by a Bacillus strain ZX99. The enzyme was a kind of ectoenzyme that could catalyze starch into isomalto-oligosaccharide effectively, but could not act on pullulan as substrate. The strain Bacillus ZX99 was mutated by ultraviolet ray and a mutant strain BS3.232 was screened. The activity of the neoamylase produced from BS3.232 increased by 60% over that from ZX99 under the same conditions. The results of thin-layer chromatography of products from starch and pullulan catalyzed by the enzyme demonstrated that the enzyme was different from neopullulanase and can be used to produce isomaltooligosaccharide from starch, including isomaltose, panose, isomaltotriose, isomaltotetose.