Tanshinone IIA prevents acute lung injury by regulating macrophage polarization / 中西医结合学报

OBJECTIVE@#Acute lung injury (ALI) is a serious respiratory dysfunction caused by pathogen or physical invasion. The strong induced inflammation often causes death. Tanshinone IIA (Tan-IIA) is the major constituent of Salvia miltiorrhiza Bunge and has been shown to display anti-inflammatory effects. The aim of the current study was to investigate the effects of Tan-IIA on ALI.@*METHODS@#A murine model of lipopolysaccharide (LPS)-induced ALI was used. The lungs and serum samples of mice were extracted at 3 days after treatment. ALI-induced inflammatory damages were confirmed from cytokine detections and histomorphology observations. Effects of Tan-IIA were investigated using in vivo and in vitro ALI models. Tan-IIA mechanisms were investigated by performing Western blot and flow cytometry experiments. A wound-healing assay was performed to confirm the Tan-IIA function.@*RESULTS@#The cytokine storm induced by LPS treatment was detected at 3 days after LPS treatment, and alveolar epithelial damage and lymphocyte aggregation were observed. Tan-IIA treatment attenuated the LPS-induced inflammation and reduced the levels of inflammatory cytokines released not only by inhibiting neutrophils, but also by macrophage. Moreover, we found that macrophage activation and polarization after LPS treatment were abrogated after applying the Tan-IIA treatment. An in vitro assay also confirmed that including the Tan-IIA supplement increased the relative amount of the M2 subtype and decreased that of M1. Rebalanced macrophages and Tan-IIA inhibited activations of the nuclear factor-κB and hypoxia-inducible factor pathways. Including Tan-IIA and macrophages also improved alveolar epithelial repair by regulating macrophage polarization.@*CONCLUSION@#This study found that while an LPS-induced cytokine storm exacerbated ALI, including Tan-IIA could prevent ALI-induced inflammation and improve the alveolar epithelial repair, and do so by regulating macrophage polarization.

Studies on blood enrichment and anti-tumor effects of combined Danggui Buxue Decoction, Fe and rhEPO based on colon cancer-related anemia model and gut microbiota modulation / 中国天然药物

Colon cancer-related anemia (CCRA) is mainly caused by systemic inflammation, intestinal bleeding, iron deficiency and chemotherapy-induced myelosuppression in colon cancer. However, the best therapeutic schedule and related mechanism on CCRA were still uncertain. Studies on blood enrichment and anti-tumor effects of combined Danggui Buxue Decoction (DBD), Fe and rhEPO based on CCRA and gut microbiota modulation were conducted in this paper. Here, CCRA model was successfully induced by subcutaneous inoculation of CT-26 and i.p. oxaliplatin, rhEPO + DBD high dosage + Fe (EDF) and rhEPO + DBD high dosage (ED) groups had the best blood enrichment effect. Attractively, EDF group also showed antitumor activity. The sequencing results of gut microbiota showed that compared to P group, the relative abundances of Lachnospiraceae and opportunistic pathogen (Odoribacter) in ED and EDF groups were decreased. Interestingly, EDF also decreased the relative abundances of cancer-related bacteria (Helicobacter, Lactococcus, Alloprevotella) and imbalance-inducing bacteria (Escherichia-Shigella and Parabacteroides) and increased the relative abundances of butyrate-producing bacteria (Ruminococcaceae_UCG-014), however, ED showed the opposite effects to EDF, this might be the reason of the smaller tumor volume in EDF group. Our findings proposed the best treatment combination of DBD, rhEPO and Fe in CCRA and provided theoretical basis and literature reference for CCRA-induced intestinal flora disorder and the regulatory mechanism of EDF.

Monoclonal Gammopathy of Renal Significance (MGRS): Prospects for Treatment in Integrated Chinese and Western Medicine / 中国结合医学杂志

Monoclonal gammopathy of renal significance (MGRS) is a pathological state which presents with a spectrum of renal lesions. MGRS is characterized by pathogenic monoclonal immunoglobulins or light chains produced by a premalignant plasma cell or B cell clone. In view of inadequate understanding in the past, the low detection rate of MGRS often results in poor outcomes and reduces quality of life of patients. Thus, MGRS stands for a group of clinical refractory renal diseases. To date, no standard treatment strategy for MGRS is available. Current consensus suggests a clone-directed approach that aims to eradicate the offending clone, but its long-term prognosis is not clear. In this article, we discuss the diagnostic methods, highlight treatment advances, and introduce integrated Chinese and Western medicine in the management of MGRS.

Comparative study on the effects of different proportions of ginkgo ketoester and donepezil on anti-dementia based on principal component analysis and multi-attribute comprehensive index methods / 药学学报

The combination of ginkgo ketoester tablet - donepezil (GD) is a popular combination commonly used in clinic for the treatment of Alzheimer's disease. To evaluate the learning and memory improving ability of different proportions of the two drugs. We optimized the ratio of GD for treatment of dementia using a mouse model. Dementia was induced by multiple neuronal damages in mice. The experimental protocols were approved by the Animal Experimental Ethical Committee of Nanjing University of Chinese Medicine and all the procedures were strictly conducted in accordance with ethical principle of animal use and care. Morris water maze, brain hematosylin-eosin staining and the changes of the neurotransmitters and related enzymes in the plasma or brain tissues were tested to determine the effect of GD on dementia mice. The results showed that the dementia mice were significantly different from the normal group in terms of behavior, pathological sections and related indicators. Compared to the dementia mice, partial administration groups could improve learning and memory ability as well as indexes in the blood and brain tissues. Both the principal component analysis and multi-attribute comprehensive index methods were used to comprehensively evaluate the total effect of GD on anti-dementia. The results showed that the combination of two drugs at the dose of 0.5 to 1 times was in a dose-effect relationship, and the dose of 1 (the clinical equivalent) had the best treatment effect. Then based on the optimal dose, GD 1∶1 had best effect, which was consistent with the clinical use of two drugs. This provides scientific basis for more effective application of the compatibility between ketoester tablet and donepezil for modern clinic medicine.

The feasibility of chemical exchange saturation transfer imaging of myocardial creatine metabolites:a 3.0 T MR study in phantom model / 中华放射学杂志

Objective To investigate the feasibility of chemical exchange saturation transfer (CEST) imaging in the measurement of myocardial creatine (Cr) metabolites in phantom model using 3.0 T MR. Methods Five phantoms were made according to the volume percentage of Cr ranging from 10 to 50 mmol/L with an interval of 10 mmol/L. 3.0 T MR examinations with base protocol sequence,sequence with and without ECG were performed. Signal to noise,CrEST effect and Z spectra were analyzed. Comparison of signal noise ratio (SNR) among the three methods was performed using an analysis of variance. Bivariate correlations were obtained through Pearson analysis. Results Phantom studies demonstrated that different concentrations of Cr exhibited significant CEST effect with the three sequences. The SNR obtained by sequences with and without ECG were both higher than that of base sequence (both P<0.05). Moreover,no significance of SNR was found between sequences with and without ECG (P>0.05). There were positive correlation of MTR between sequences with ECG,sequences without ECG and base protocol sequence (r2= 0.974 and 0.997, both P<0.05). Conclusion Compared with base protocol sequence, the optimized sequence with ECG can acquire higher SNR CrEST images,indicating that myocardial CrEST imaging could be performed in clinical practice.

Application of dual-source CT combined with intelligent modulation and iterative reconstruction in aortic dissection imaging / 中华放射医学与防护杂志

Objective To explore the clinical application value of second-generation dual-source CT combined with intelligent modulation and iterative reconstruction in emergency aortic dissection imaging.Methods A total of 40 emergency patients with clinical suspected aortic dissection were included in this study.Conventional scanning was performed in the control group,and large-pitch intelligent modulation and iterative reconstruction were performed in the test group.The mean CT value,mean noise,signal noise ratio(SNR),contrast noise ratio(CNR),effective dose,image quality and aortic root image quality were evaluated and analyzed.Results Totally 40 patients successfully completed CT aortic dissection imaging.There was no difference in image quality between the two groups (P＞ 0.05).The quality of aortic root images in the test group was better than that in the control group,and the difference was statistically significant(x2=22.556,P＜0.05).The mean CT value and mean noise of aorta in the control group were slightly higher than those in the test group.However,SNR and CNR in the test group were higher than those in the control group,and the difference was statistically significant (t =-21.042,-15.924,8.530,11.495,P＜0.05).The effective dose of the control group [(10.59±3.89)mSv] was significantly higher than that [(6.39±0.81) mSv] of the test group,the difference was statistically significant (t =-12.327,P＜0.05).Conclusions The combined intelligent modulation technique and iterative reconstruction technique with dual-source CT large pitch scanning can meet the requirements of image quality and reduce the effective dose,and can be used as a conventional imaging method for emergency CT of aortic dissection.

Effect of adenosine on autophagy and proliferation of hepatocellular carcinoma cells / 中国药理学通报

Aim To investigate the effect of adenosine on the autophagy and proliferation of hepatocellular carcinoma cells, and improve the curative effect of a-denosine on hepatocellular carcinoma. Methods HepG2 cells were incubated with adenosine, CCK-8 method was used to study the changes of cell prolifera-tion,Western blot was used to study the expression of LC3-Ⅱ and LC3-Ⅰ, and MDC staining was used to observe the number of autophagosomes. Results HepG2 cells were incubated with adenosine(1.0~4.0 mmol·L-1) for 48 h,the proliferation of HepG2 cells were detected at the different time points (12,24,48 h),and the result showed the proliferation was signifi-cantly inhibited by adenosine (P < 0.01). HepG2 cells were incubated with adenosine (0.2,0.5,1.0, 2.0,4.0 mmol·L-1) for 24 h,the ratio of LC3-Ⅱ/LC3-Ⅰ decreased significantly in low concentration of adenosine group (0.2, 0.5 mmol·L-1, P <0.05;1.0 mmol·L-1,P<0.01),and the ratio of LC3-Ⅱ/LC3-Ⅰ increased significantly in higher concentration of adenosine group (4.0 mmol·L-1, P <0.05). HepG2 cells were incubated with adenosine(1.0 mmol·L-1) for 24 h, the ratio of LC3-Ⅱ/LC3-Ⅰ de-creased significantly at 6,12 and 24 h detecting point, the number of autophagosomes were reduced, the low-est ratio of LC3-Ⅱ/LC3-Ⅰ and autophagosomes were observed at 12 h detecting point(P<0.01). Conclu-sions Adenosine inhibits the proliferation of hepato-cellular carcinoma cells,the low concentration of aden-osine inhibits the autophagy,while the high concentra-tion of adenosine increases the autophagy, which is of great significance to reduce multi-drug resistance and improve the therapeutic effect of anti-hepatoma drugs.

Spatial distribution of radiation dose field from mobile CT head scanning / 中华放射医学与防护杂志

Objective To identify the spatial distribution of stray radiation from mobile CT head scanning for the purpose of radiation protection.Methods The head series of CareTom mobile CT were scanned and the radiation dose was measured using TLD (LiF:Mg,Cu,P).The isodose maps of radiation dose field were plotted using Matlab software.Results Radiation dose in the front of the mobile CT was slightly higher than that in the back.The maximum value of 0.255 mGy was found to be at 0.5 m from the scanning hole center.Conclusions The stray radiation dose from mobile CT head scanning was relatively low.However in order to avoid the damage to the operators and other medical workers from long-term low dose exposure,it should keep 2 m away from mobile CT,beside or behind,when in operation.

The clinical value of anteroposterior and lateral scout image combined with Care Dose 4D and Care kV in reducing radiation dose of chest CT scanning / 中华放射医学与防护杂志

Objective To investigate the feasibility and clinical value of anteroposterior and lateral scout scan combined with Care Dose 4D and Care kV in chest CT scan.Methods A total of 60 patients of clinical diagnosis with lung tumor were enrolled.Those patients were randomly divided into test group and control group.Control group underwent a scan protocol with lateral scout scan combined with Care Dose 4D and Care kV,while anteroposterior and lateral scout scan combined with Care Dose 4D and Care kV were performed in test group.The signal-to-noise ration (SNR),contrast-to-noise ratio (CNR),and overall image quality of two groups of images and diseased tissues were analyzed and evaluated by two high-grade radiologists using double-blind method.Effective doses (E) were also calculated.Results All the 60 patients had successfully completed the chest CT scans.Test group overall image quality (4.57 ± 0.45) and control group overall image quality (4.73 ± 0.45) had no statistically significant difference (P ＞ 0.05).The control group image SNR,CNR and diseased tissue SNR,CNR compared with test group had no statistical significance difference (P ＞ 0.05).The difference of the volume CT dose index (CTDIvol),dose-length product (DLP) and effective dose (E) of test group and control group was statistically significant (t =8.514,8.464,8.464,P ＜ 0.001).Compared with control group,the effective dose of test group decreased by 33.3％.Conclusions Compared with lateral scout scan,the technology of anteroposterior and lateral scout scan combined with Care Dose 4D and Care kV can decrease radiation dose without reducing the image quality.This technology can therefore be considered as a regular imaging modality for chest CT scan.

HPLC method to assay the content of psoralen and angelicin in Zhuanggu granules / 药学实践杂志

Objective To develop a HPLC method for psoralenand angelicin content assay in Zhuanggu granules .Meth-ods Agilent Zorbax C18 column(4 .6 mm × 250 mm ,5 μm) was used with methanol and water (45∶55) as mobile phase .The detection wavelength was set at 245 nm .The flow rate was 1 .0 ml/min and column temperature at 25 ℃ .The injection volume was 10 μl .Psoralenand angelicin were extracted with 70% ethanol under water bath .Results The linearity was obtained over 3 .75~40 μg/mlfor psoralenand angelicin .The RSDs were less than 2% .The average recovery was between 94% and 105% . Conclusion This simple and accurate HPLC method was suitable for the quality control of Zhuanggu granules .

Long-term exposure to PM2.5 from automobile exhaust results in reproductive dysfunction in male rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effects of long-term exposure to particulate matter 2.5 (PM2.5) from automobile exhaust on the reproductive function of Sprague Dawley (SD) rats.</p><p><b>METHODS</b>Forty-five male SD rats, weighing 80 - 94 g and aged 28 days, were randomly assigned to receive intra-tracheal administration of 0.9% normal saline (control group, n = 15), PM2. 5 at 2 μg per 100 g body weight per day (low-dose PM2.5 group, n = 15), and PM2.5 at 16 μg per 100 g body weight per day (high-dose PM2.5 group, n = 15), qd, for 60 successive days. After the last 24-hour exposure, 10 rats were taken from each group for copulation with normal female ones, while the others were sacrificed, their testes removed for sperm count and deformity, pathological examination, and determination of the Connexin43 expression.</p><p><b>RESULTS</b>The conception rate was significantly decreased in the low- and high-dose PM2.5 groups as compared with that of the control (70% and 50% vs 100%), and so were the sperm count and quality. The rats in the PM2.5-exposed groups showed significantly disordered histological structure of the seminiferous tubules, reduced sperm count in the testicular lumen, some exfoliated secondary spermatocytes, downregulated Connexin43 expression in the testis, and damaged blood-testis barrier.</p><p><b>CONCLUSION</b>Long-term exposure to PM2.5 from automobile exhaust damages the reproductive function of male SD rats.</p>

Analysis of cell morphology and immunophenotypic characteristics in 47 cases of multiple myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was to investigate the cell morphology and cell immune phenotypic characteristics in patients with multiple myeloma (MM).</p><p><b>METHODS</b>The flow cytometry with multiparametric direct immunofluorescence technique, and CD45/SSC and CD38(+)(+)/CD138(+) gating were used to measure cell markers CD138, CD38, CD56, CD117, CD3, CD13, CD33, CD19, CD7, CD20, CD22, CD34, CD28 in 47 MM patients. At the same time the morphology examination of bone marrow cells was performed.</p><p><b>RESULTS</b>The suspicious myeloma cell ratio in MM patients was 9.42%-74.25% detected by flow cytometry, moreover, the myeloma cell ratio detected by morphology examination was 11.0%-80.6%, there was a good correlation between the two detection methods (r(2) = 0.54, P < 0.001). The ratio of antigen positive expression was as follows: 74.46% for CD138, 100% for CD38, 57.44% for CD56, 40.42% for CD117, 6.38% for CD13, 19.15% for CD33, 8.51% for CD20, 27.66% for CD28, 2.12% for CD22, 4.25% for CD34, 0% for CD3, 0% for CD19, 0% for CD7.</p><p><b>CONCLUSIONS</b>CD45/SSC and CD38(+)/CD138(+) gating technique can accurately gate multiple myeloma cell sets which need analysis, the majority of myeloma cells expreses CD138, CD38, CD56 antigens. The immunophenotypic analysis combined with the cell morphology examination more contribute to the diagnosis and differential diagnosis of multiple myeloma.</p>

Effects of Wenxiao Decoction on the expression of interleukin-6, intercellular adhesion molecular-1 and monocyte chemoattractant protein-1 in experimental atherosclerotic rabbits / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effects of different doses of Wenxiao Decoction on the expression of interleukin-6 (IL-6), intercellular adhesion molecule-1 (ICAM-1), and monocyte chemoattractant protein-1 (MCP-1) in experimental atherosclerotic rabbits and to explore the mechanism by which it alleviates atherosclerosis.</p><p><b>METHODS</b>Sixty New Zealand rabbits were randomly divided into six groups: a blank group, a model group, a Simvastatin group, and high-, medium-, and low-dosage Wenxiao Decoction groups. Except for those in the blank group, all rabbits were fed with a high-cholesterol diet. Carotid atherosclerosis was established by balloon-induced carotid artery endothelium injury in conjunction with the high-cholesterol diet. After 8 weeks, all animals were euthanized to evaluate levels of IL-6 and ICAM-1 expressions (by enzyme linked immunosorbent assay) and of MCP-1 (by immunohistochemistry staining).</p><p><b>RESULTS</b>The expressions of IL-6, ICAM-1, and MCP-1 were significantly increased in all groups except the blank group (P<0.05). However, the rabbits in the Wenxiao Decoction groups and the Simvastatin group showed significantly lower levels of IL-6, ICAM-1, and MCP-1 expression than those in the model group (P<0.05). The expressions of IL-6, ICAM-1, and MCP-1 in the highdosage Wenxiao Decoction group and the Simvastatin group were lower than those in the low-dosage Wenxiao Decoction group (P<0.05). The expression of MCP-1 in medium-dosage Wenxiao Decoction group was lower than that in the low-dosage group (P<0.05).</p><p><b>CONCLUSIONS</b>High, medium, and low doses of Wenxiao Decoction can inhibit the expressions of IL-6, ICAM-1, and MCP-1, which may prevent and stabilize atherosclerotic plaques. There may be a direct relationship between dosage and therapeutic efficacy of Wenxiao Decoction.</p>

Expression of indoleamine 2, 3-dioxygenase and its correlation with prognosis in breast cancer patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression of indoleamine 2, 3-dioxygenase (IDO) in breast cancer and its correlation with clinicopathologic factors and prognosis.</p><p><b>METHODS</b>The expression of IDO, CD31, CD105 proteins in 40 specimens of breast cancer were assessed by immunohistochemistry.</p><p><b>RESULTS</b>The overexpression rate of IDO in breast cancer was 67.5% (27/40), and expression of IDO was closely associated with clinical stage and lymph nodes metastasis. The disease-free survival rate in patients with IDO overexpression was not significantly lower than that in patients with negative or low expression of IDO (P > 0.05). Moreover, the expression of IDO was positively correlated with CD105-labeled microvessel density (r = 0.659, P < 0.05).</p><p><b>CONCLUSIONS</b>Expression of IDO is associated with clinical stage and lymph nodes metastasis, and microvessel densitty. IDO expression may promote the growth and metastasis of breast cancer, probably via the increased agiogenesis. A larger sample study is needed to verify whether the prognosis of beast cancer is significantly correlated with IDO expression.</p>

Up-regulation of the expression of S100A8 and S100A9 in lung adenocarcinoma and its correlation with inflammation and other clinical features / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>S100A8 and S100A9 are two members of the S100 protein family characterized by the presence of two Ca2+-binding sites of the EF-hand type. Previous studies suggested that the whole S100 family displays significant functions in tumor growth, progression and invasion. This study aimed to determine the expression of the two indices of the family, S100A8 and S100A9, in lung cancer tissues and normal lung tissues and its correlation with clinical features.</p><p><b>METHODS</b>A total of 60 cases with a variety of clinical data that were diagnosed with different histological subtypes of lung cancer were investigated. Semi-quantitative reverse transcriptase-PCR (Sq-Rt-PCR) and immunohistochemical staining of cancer, adjacent and peripheral lung tissues were executed to distinguish the expression patterns of S100A8 and S100A9 and to further clarify their correlation with clinical features.</p><p><b>RESULTS</b>Immunohistochemical staining of both proteins showed a significant up-regulation in lung cancer tissue (S100A8, S100A9, P<0.0001), and PCR revealed that the levels of S100A8 and S100A9 expression were significantly higher in lung cancer tissues (S100A8 P=0.002/0.004; S100A9 P=0.022/0.026). The higher expression was found to be correlated with the clinical characteristics of adenocarcinoma, inflammation and stage IV lesion.</p><p><b>CONCLUSIONS</b>S100A8, S100A9 up-regulation was found in the lung adenocarcinoma and end stage lung cancer tissue, the correlation of which with their higher expression in inflammatory lung tissues may indicate the collaborative effect of inflammation on the progression of cancer.</p>

Alloreactive NK cells enhance the effect of donor lymphocyte infusion in the management of relapsed lung cancer after haploidentical hematopoietic stem cell transplantation / 中国实验血液学杂志

This study was aimed to study the potential effects of alloreactive NK cells (allo-NKs) in therapy of relapsed lung cancer after haploidentical hematopoietic stem cell transplantation using donor lymphocyte infusion (DLI). The F1 donors derived-NK cells were purified with MACS magnetic separation system, in which the proportion of the alloreactive Ly49A(+) cells was detected by flowcytometry and alloreactivity was measured by LDH method. The relapse model of lung cancer after haploidentical-HSCT was established. The distribution kinetic of infused donor lymphocytes in vivo was analyzed. The inhibition of relapse tumor, infiltration of lymphocytes in situ and fluctuation of 22 kinds of cytokines in serum after DLI were compared among different groups. The results showed that the infused donor cells of allo-NK group were accumulated mostly in lung, spleen and kidney for more than 48 hours with considerable higher levels according to the distribution kinetic curve. The sizes of relapse tumors between chemotherapy + PBS group and chemotherapy + DLI group showed no difference. However, the relapsed tumors in allo-NK + DLI group were significantly smaller than that in chemotherapy + DLI group or allo-NK + PBS group, in which increased infiltration of lymphocytes were defined in situ. The levels of cytokines such as MCP-1, IL-17, IL-12 and MCP-5 in serum of allo-NK + DLI group ascended compared with control group, though the level of IL-10 declined simultaneously. It is concluded that allo-NKs prolong the survival time of infused donor lymphocytes in vivo, promote the secretion of inflammatory cytokines and Th1-type of cytokines, and further improve the antitumor effects of DLI against relapse after transplantation.

The anti-tumor activity of GM-CSF-modified lung cancer cell vaccine and its synergism in combination with chemotherapy / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the anti-tumor effects and mechanism of tumor vaccines and whether chemotherapeutic agents administered prior to immunotherapy could augment the efficacy of the vaccines.</p><p><b>METHODS</b>C57/BL mice inoculated with Lewis lung cancer cells were used as tumor models. Granulocyte-macrophage colony-stimulating factor (GM-CSF) gene modified LA795 and Lewis lung cancer cell lines were administered as allogeneic and autologous tumor vaccines, respectively. After Lewis cells (1 x 10(7)) inoculation, the mice received irradiated GM-CSF secreting cancer vaccine solely or in combination with carboplatin. The survival of the mice was observed. The cytotoxicity of spleen cells or purified CD8(+) cells was analyzed by lactate dehydrogenase (LDH) assay. Serum level of IL-4 and IFN-gamma was detected using ELISA method.</p><p><b>RESULTS</b>The cytotoxicity of the spleen cells or purified CD8(+) T cells against Lewis cells in the mice immunized with cancer cell vaccine was significantly increased, relative to that of the control, untreated group (P < 0.05). Serum level of Th1-type cytokine IFN-gamma was increased after vaccination, whereas Th2-type cytokine IL-4 showed no significant change. The GM-CSF secreting cancer cell vaccine had no significant influence on the survival of the mice with established heavy tumor burden. The combination of chemotherapy and cancer vaccine could statistically prolong the survival time; whereas any method itself had no significant effect.</p><p><b>CONCLUSION</b>The GM-CSF secreting cancer cell vaccine can induce immune responses. The chemotherapeutic agents may be beneficial to enhance the anti-tumor activity of cancer vaccine.</p>

Application of low dose of fludarabine and cyclophosphamide combined with donor NK Cells as a non-myeloablative conditioning regimen for the haploidentical hematopoietic stem cell transplantation in mice / 中国实验血液学杂志

This study was aimed to investigate the feasibility of low dose of fludarabine, cyclophosphamide combined with donor derived alloreactive NK cells as a new nonmyeloablative conditioning regimen in the haploidentical hematopoietic stem cell transplantation (haploidentical HSCT). F1 derived-NK cells were enriched with MACS magnetic separation system, in which the proportions of the Ly49C+ and Ly49A+ cells were detected by flow cytometry and the alloreactivity was measured by LDH method. The haploidentical HSCT models were constructed, and the myeloablativity in vivo, donor engraftment and the intensity of GVHD were compared between different myeloablative and nonmyeloablative conditioning regimens, including 9 Gy TBI, 6.5 Gy TBI, flu + cy, and flu + cy + allo-NK. The results showed that the flu + cy + allo-NK conditioning was nonmyeloablative, but the rate of donor chimerism after haploidentical HSCT was significantly higher as compared with other nonmyeloablative methods, which were (28.70 +/- 5.90)% in bone marrow and (46.40 +/- 5.00)% in spleen at day 21 post-transplantation. When compared with the flu + cy conditioning, the intensity of GVHD was slight in the flu + cy + allo-NK group, in which only a half of C57BL/6 recipients experienced weight loss, and no distinct pathological damages observed in the liver, intestine, kidney and skin samples. It is concluded donor derived-alloreactive NK cells can facilitate engraftment of the haploidentical hematopoietic stem cells and mitigate GVHD. The flu + cy + allo-NK conditioning provides a new method for those elder patients with high-risk solid tumor undergoing haploidentical-HSCT.

Elevation of tumor necrosis factor-alpha, interleukin-1beta and interleukin-6 levels in aortic intima of Chinese Guizhou minipigs with streptozotocin-induced diabetes / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Large animal models with toxin-mediated pancreatic damage have been used extensively in researches with respect to diabetes mellitus and cardiovascular diabetic complications. The present study aimed to establish Chinese Guizhou minipig models with streptozotocin (STZ)-induced diabetes and characterize the animal models by analyzing inflammatory cytokine levels in aortic wall, such as tumor necrosis factor (TNF)-alpha, interleukin-1beta (IL-1beta) and interleukin-6 (IL-6).</p><p><b>METHODS</b>Twenty-two male Chinese Guizhou minipigs (age, 4 to 6 months; weight, 20 kg to 30 kg) were divided into STZ-induced diabetic group (n = 12) and control group (n = 10). STZ (125 mg/kg) was administrated to induce hyperglycemia and afterwards insulin was used to control fasting blood glucose levels below 10 mmol/L. Oral glucose tolerance test (OGTT) was performed before and one month after STZ administration and serum concentrations of alanine transaminase, asparagine transaminase, albumin, blood urea nitrogen, creatinine, lipids and white blood cell count were measured before and six months later. Animals in both groups were euthanized after six months and pancreas was examined immunohistochemically for islet beta cells. Aortic intima of diabetic minipigs and controls was analyzed for TNF-alpha level in tissue conditioned medium by Western blot. TNF-alpha, IL-1beta and IL-6 mRNA levels in aortic intima were assayed by reverse transcription and polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Significant elevation in serum glucose levels was observed one month and six months after STZ induction (P < 0.001) and markedly increased OGTT values were noted, compared with baseline data. The normal pancreas had many irregular sized islets and small clusters of islet beta cells, while in pancreas of diabetic minipigs islet beta cells almost disappeared. No statistical difference was notified in serum concentrations of biochemical examinations before and six months after STZ induction. Western blot demonstrated dramatically increased TNF-alpha level in aotic intima conditioned medium, and significant elevation of TNF-alpha, IL-1beta and IL-6 mRNA levels was revealed by RT-PCR.</p><p><b>CONCLUSIONS</b>The present study has established Chinese Guizhou minipig models with STZ-induced diabetes. Inflammatory cytokines (TNF-alpha, IL-1beta and IL-6) significantly elevated in aortic intima of diabetic minipigs.</p>

Anti-leukemia immunity induced by dendritic cells fused with L615 tumor cells / 中国实验血液学杂志

This study was aimed to investigate the specific anti-L615 leukemia cell immunity induced by L615/DC fused cell vaccine in vivo and in vitro. BM-derived DCs were generated from bone marrow of 615 mice by culturing for 9 - 10 days in culture medium supplemented with GM-CSF and IL-4. Irradiated L615 tumor cells were fused with DC by using PEG to form fused cell vaccine, with which 615 mice were immunized. After immunization, the specific proliferation ability and cytotoxicity against L615 leukemia cells in vitro were examined by MTT and LDH methods. Anti-leukemia effect of fused cell vaccine in vivo was studied by observing the immunotherapy effects on L615 tumor-bearing mice. The results showed that fully mature and functional bone marrow-derived DC were obtained. L615/DC fused cell vaccine could elicit potent specific proliferation response of spleen T cells from immunized mice when contacting with the same antigen at the second time, and could also elicit the effective cytotoxic activity against L615 leukemia cells in vitro, which were significantly different from other groups. In vivo the average survival time of the tumor-bearing mice received immunotherapy with L615/DC fused cell vaccine was 25.7 +/- 1 days, and one fourth of treated tumor-bearing mice survived for long time, but the mice of control group died all, their average of survival time was 17.5 +/- 1 days. The immunized mice survived with no evidence of recurrence when exposed to the second attack of lethal dose of living L615 cells 2 months later. It is concluded that L615/DC fused cell vaccine can improve the immunogenecity of L615 and induce effectively the specific anti-leukemia immunity against L615 leukemia cells to eliminate the residual leukemia cells, prolong the survival time and induce the immune memory to avoid the relapse. Thus, the fused cell vaccine may be an attractive strategy for malignance immunotherapy.