Construction and preliminary application of drug carrier system based on yeast membrane vesicles / 药学学报

As an edible eukaryotic microorganism, Saccharomyces cerevisiae has the characteristics of high safety, rapid proliferation, low cost, easy transformation, etc. It has been widely used to produce vaccines, antibodies, insulin, etc. Up to now, yeast components, such as cell wall and yeast microcapsules, have been widely used in the treatment of tumors, inflammatory virus infection, post-traumatic osteoarthritis and other diseases. Among them, the components of yeast cell membrane are relatively simple and stable, which are easy to be extracted on a large scale. Therefore, yeast cell membrane material was used to construct yeast membrane vesicle nanosystem, and its biomedical application was preliminarily explored. In this study, Saccharomyces cerevisiae membrane vesicle (SMV) was prepared by co-extrusion method, and the particle size and surface potential of SMV, drug loading and release characteristics, stability, cell safety, and in vitro therapeutic effect were investigated. The results showed that the average particle size of SMV was 185.1 nm. Curcumin and silica nanoparticles were effectively encapsulated by co-incubation and ultrasonic methods, and the characteristics of cell membrane proteins were maintained. Moreover, SMV had good stability and biocompatibility. In addition, SMV could be effectively uptaken by macrophages RAW 264.7, and curcumin loaded SMV could effectively eliminate reactive oxygen species (ROS). In conclusion, the yeast plasma membrane vesicles prepared in this study could effectively deliver curcumin drugs and encapsulate nanoparticles, and could be effectively absorbed by macrophages and effectively eliminate ROS, providing new ideas and new methods for biomedical applications of yeast membrane materials.

Echocardiographic Measurements Results in Healthy Han, Tibetan and Yugur Adults / 中国循环杂志

Objectives:The aim of this study was to compare the potential differences on echocardiographic cardiac chambers size and systolic function among the healthy Han, Tibetan and Yugur adults. Methods:A total of 1 820 healthy volunteers aged 20 to 80 years were screened at Qinghai province and Gansu province, including 887 Han (456 men), 506 Tibetan (262 men) and 427 Yugur (187 men) Chinese adults. Standard two-dimensional echocardiography was performed to obtain measurements of the cardiac chambers and left ventricular ejection fraction (LVEF). The impacts of nationality, age and gender on all echocardiographic results were analyzed. Results:The values of left ventricular end-diastolic volume was higher in Yugur male adults than in Han and Tibetan male adults ([96.29±21.60]ml vs [93.64±18.30] ml vs [91.16±16.77] ml, P＜0.05).The differences remained after adjusting body surface area ([54.16±11.14]ml/m2 vs [53.92±10.20] ml/m2 vs [52.76±8.94] ml/m2, P＜0.05). There was no significant difference in LVEF among three nationalities (P＞0.05). The values of cardiac chambers dimensions were significantly higher in men than in women (P＜0.05). The anteroposterior dimension of the left atrium increased with age in both genders (P＜0.05). Conclusions:Our results show that the left ventricular end-diastolic volume is related to nationality and some parameters vary with gender and age. Not only the differences in age, gender, body size, but also ethnic differences should be considered when defining the normal values of echocardiographic examination.

Effect of Cordyceps sinensis powder on renal oxidative stress and mitochondria functions in 5/6 nephrectomized rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Cordyceps sinensis (CS) powder on renal oxidative stress and mitochondria functions in 5/6 nephrectomized rats, and to primarily explore its possible mechanisms.</p><p><b>METHODS</b>Totally 30 male Sprague-Dawley rats were divided into the sham-operation group, the model group, and the treatment group by random digit table, 10 in each group. A chronic kidney disease (CKD) rat model was prepared by one step 5/6 nephrectomy. Rats in the treatment group were intragastrically administered with CS powder solution at the daily dose of 2 g/kg, once per day. Equal volume of double distilled water was intragastrically administered to rats in the sham-operation group and the model group. All medication lasted for 12 weeks. The general condition of rats, their body weight, blood pressure, 24 h proteinuria, urinary N-acetyl-β-D-glucosaminidase (NAG), serum creatinine (SCr) , and blood urea nitrogen (BUN) were assessed before surgery, at week 2, 4, 6, 8, 10, and 10 after surgery. Pathological changes of renal tissues were observed under light microscope. Morphological changes of mitochondria in renal tubular epithelial cells were observed under transmission electron microscope. Activities of antioxidant enzymes including reduced glutathione (GSH), manganese superoxide dismutase (MnSOD), and malondialdehyde (MDA) in fresh renal tissue homogenate were detected. Mitochondria of renal tissues were extracted to detect levels of mitochondrial membrane potential and changes of reactive oxygen species (ROS). And expressions of cytochrome-C (Cyto-C) and prohibitin in both mitochondria and cytoplasm of the renal cortex were also measured by Western blot.</p><p><b>RESULTS</b>(1) Compared with the sham-operation group, body weight was significantly decreased at week 2 (P <0. 01), but blood pressure increased at week 4 (P <0. 05) in the model group. Compared with the model group, body weight was significantly increased at week 12 (P <0. 01), but blood pressure decreased at week 8 (P < 0. 01) in the treatment group. (2) Compared with the sham-operation group, 24 h proteinuria, urinary NAG, blood SCr and BUN significantly increased in the model group (all P <0. 01). Compared with the model group, blood and urinary biochemical indices all significantly decreased in the treatment group (all P <0. 01). (3) Results of pathological renal scoring: Glomerular sclerosis index, scoring for tubulointerstitial fibrosis, degree of tubulointerstitial inflammatory infiltration were all obviously higher in the model group than in the sham-operation group (all P <0. 01). All the aforesaid indices were more obviously improved in the treatment group than in the model group (all P <0. 01). (4) Compared with the sham-operation group, activities of MnSOD and GSH-Px were significantly reduced, but MDA contents obviously increased in the renal cortex of the model group (all P <0. 01). Compared with the model group, activities of MnSOD and GSH-Px obviously increased (P <0. 05, P <0. 01), but MDA contents obviously decreased in the renal cortex of the treatment group (P <0. 01). (5) Compared with the sham-operation group, the mitochondrial membrane potential significantly decreased, but ROS levels significantly increased in the model group (all P <0.01). Compared with the model group, mitochondrial transmembrane potential increased in the treatment group, thereby inhibiting the tendency of increased production of ROS (both P < 0. 01). (6) Results of Western blot showed that, compared with the sham-operation group, expression levels of mitochondrial Cyto-C and Prohibitin were significantly reduced in the renal cortex (P <0. 01), but significantly elevated in the cytoplasm of the model group (P <0. 01). Compared with the model group, each index was obviously improved in the treatment group with statistical difference (P <0. 05, P <0. 01).</p><p><b>CONCLUSION</b>CS powder had renal protection, and its mechanism might partially depend on in- hibition of oxidative stress and protection for mitochondria.</p>

Inhibition of resvertrol on osteoblast-like trans-differentiation of vascular smooth muscle cells / 中草药

Objective: To investigate the inhibition of resvertrol (Res) on angiosteosis, and to observe the effects of Res at different concentration on osteogenic conversion of human umbilical artery vascular smooth muscle cells (hVSMCs) induced by elevated calcium (Ca2+) and phosphate (P) culture. Methods: The hVSMCs were cultured in vitro, and identified by morphology and α-smooth muscle actin (α-SMA) antibody. High calcium (2.5 mmol/L) and high phosphate (3.0 mmol/L) culture was used to induce the osteogenic differentiation of hVSMCs. The calcification was confirmed by Alizarin red staining. Cells were divided into five groups such as normal control (NC, normal concentration of Ca2+ and P), positive control (PC, high Ca2+ and high P), and three Res (5, 10, and 20 μmol/L) groups. Calcium deposition in cells was measured by Arsenazo III after 12 d. Real-time PCR and Western blotting were used to detect mRNA and protein expression of BAP, BMP2, and OPN. Results: Compared to NC group, calcium deposition in the cells dramatically increased by five times in PC group, the protein expression of α-SMA decreased, and the mRNA and protein expression of BAP, BMP-2, and OPN obviously increased (P < 0.05). Res at different concentration could reduce the cellular Ca2+. The levels of BAP, BMP2, and OPN were highest in PC group than NC and three Res groups. Furthermore, the expression of BMP2 and OPN decreased with increasing Res concentration. Conclusion: Res prevents the process of the osteogenic differentiation of hVSMCs induced by high calcium and high phosphate. The degree of osteogenic differentiation is falling with increasing Res concentration.

Treatment of unstable middle 1/3 of clavicular fractures with composite external fixator in adult patients / 中国骨伤

<p><b>OBJECTIVE</b>To explore methods and clinical effects of composite external fixtor in treating adult patients with unstable middle 1/3 of clavicular fractures.</p><p><b>METHODS</b>From March 2008 to March 2011,36 patients with unstable middle 1/3 of clavicular fractures were treated with composite external fixtor. There were 24 males and 12 females, aged from 18 to 55 years old with an average of 43 years old. Twenty cases on the left side and 16 cases on the right side. Time from injury to operation was 2 to 6 days (averaged 3.5 days). According to Robinson classification, there were 7 cases with type 2A2, 18 cases with type 2B1, and 11 cases with type 2B2. No vessels and nerve damage occurred before opreation. The clinical effects were evaluated according to Neer scoring.</p><p><b>RESULTS</b>All cases were followed up from 6 to 12 months with an average of 8 months. The mean Neer score was 88.3 +/- 6.2, which included pain 31.6 +/- 3.2, functional score 25.7 +/- 2.2, range of motion score 21.1 +/- 1.7, and anatomy score 8.8 +/- 0.8. There were 22 cases in excellent, good in 13, fair in 1. Two cases occurred pin tract infection.</p><p><b>CONCLUSION</b>Composite external fixtor is an optional method in treating unstable middle 1/3 of clavicular fracture, and can obtain a good clinical effects.</p>

Neuroendocrine neoplasm of digestive system with different grades: a clinicopathologic and prognostic study / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic and prognostic features of neuroendocrine neoplasm of digestive system with different grades.</p><p><b>METHODS</b>The clinicopathologic features of 139 cases of neuroendocrine neoplasm occurring in digestive system were retrospectively reviewed and graded according to the 2010 World Health Organization classification of tumours of the digestive system. Immunohistochemical study for synaptophysin, chromogranin A and Ki-67 was carried out. The follow-up and survival data were analysed using Kaplan-Meier method. Prognostic factors were tested by Log-rank testing and independent risk factors were analysed using Cox regression model.</p><p><b>RESULTS</b>Amongst the 139 cases studied, there were 88 cases (63.3%) of grade 1 tumors, 9 cases (6.5%) of grade 2 tumors and 42 cases (30.2%) of grade 3 tumors. There was diffusely positive staining for synaptophysin and chromogranin A in most of the grade 1 and grade 2 tumors. The staining in grade 3 tumors however was focal (P < 0.05). The differences in tumor size, depth of invasion, presence of tumor emboli, perineural permeation, nodal involvement, distant metastasis and survival rate amongst the three groups was statistically significant (P < 0.05).</p><p><b>CONCLUSIONS</b>There is significant difference in the clinicopathologic and prognostic features of neuroendocrine neoplasm of digestive system with different grades. It is considered as an independent prognostic factor and represents a useful tool for prognostic evaluation of such tumors, both in clinical practice and research.</p>

Clinicopathologic significance of bcl-6 gene rearrangement and expression in three molecular subgroups of diffuse large B-cell lymphoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the role of bcl-6 gene rearrangement and bcl-6 expression in three molecular subgroups of diffuse large B-cell lymphoma (DLBCL) and its clinicopathological significance.</p><p><b>METHODS</b>Tissue microarray including 163 newly diagnosed DLBCL was constructed. Fluorescence in situ hybridization (FISH) was performed to detect the bcl-6 gene rearrangement and immunohistochemistry (EnVision method) was used to evaluate the expression of bcl-6, Ki-67, cyclin D3, Geminin and P27(Kip1) proteins in DLBCL. The association with clinicopathological features was analyzed.</p><p><b>RESULTS</b>One hundred and forty nine of 163 cases were further classified into three molecular subgroups: 40 cases of germinal center B-cell-like (GCB) type, 75 cases of activated non-germinal center B-cell-like (ABC) type, 34 cases of Type 3. Of these 149 cases, FISH for bcl-6 gene rearrangement was successful in 118 cases. bcl-6 gene rearrangement was observed in 33 of 118 (28.0%) cases. The bcl-6 gene rearrangement was more frequently seen in the ABC subgroup (22/62, 35.5%) than in GCB (6/31, 19.4%) and Type 3 subgroups (5/25, 20.0%, P=0.16). The correlation of bcl-6 gene rearrangement and expression of its encoded protein was further analyzed. Most of DLBCL (26/33, 78.8%) with bcl-6 gene rearrangement presented with overexpression of its encoded protein, which was higher than those without bcl-6 gene rearrangement (53/84, 62.4%, P=0.088). DLBCL with bcl-6 gene rearrangement (24/33, 72.7%) more frequently expressed cyclin D3, and had a higher proliferative activity than those without bcl-6 gene rearrangement (37/81, 45.7% , P=0.009). Twenty-nine of 33 (87.9%) cases of DLBCL with bcl-6 gene rearrangement presented with advanced stage (Ann Arbor stage III/IV), which was higher than those without bcl-6 gene rearrangement (65/85, 76.5% , P=0.167). Univariate Cox proportional hazards regression analysis showed that bcl-6 gene rearrangement was associated with an increased relative risk (at 1.842) of death in DLBCL cases compared with those without bcl-6 gene rearrangement.</p><p><b>CONCLUSION</b>Overexpression of bcl-6 protein caused by bcl-6 gene rearrangement may play some important roles in the development and/or progression of a subset of DLBCL.</p>

Determination of CD34(+) cells in mobilized peripheral blood by flow cytometry / 中国实验血液学杂志

To explore a simple and effective method to determinate the volume of CD34(+) cells in the peripheral blood of donors received drug mobilization for stem cell transplantation by using flow cytometry, the mobilized peripheral blood from donors and 100 micro l fresh whole blood were labeled with monoclonal antibodies Anti-CD34-PE and Anti-CD45-FITC, after lying the red blood cells, and assessed with flow cytometer FL2 (log) vs SSC (log) and FL1 (log) vs SSC (log) were mainly used for analysis windows. The results showed that a level of CD34(+) cells in whole nucleated cells as low as 0.05% - 0.1% can be detected effectively using this method when 10(5) nucleated cells were counted. At day 5 or day 6, the level of CD34(+) cells in most samples of patients reached a peak volume, some of samples and the levels were more than one percent in. It was concluded that CD34(+) cells can be effectively determined by using this method. According to the relative rate of CD34(+) cells, the time to harvest the stem cells in blood can be determined.

The number and function analysis of T cell from mice immunized with MIF gene-modified tumor vaccine / 第二军医大学学报

Objective: To investigate the role of T cell in the antitumor immune responses induced by MIF gene-modified tumor vaccine. Methods: MIF gene was transferred into FBL3 erythroleukemia cel l by adenovirus carrier and a new type of tumor vaccine was prepared. The chang es of the number and the function of T cell in spleen and lymph node was observe d. Results: After the mice were immunized with MIF gene-m odified FBL3 vaccine, the number of lymphocyte in spleens and lymph nodes increa sed markedly and the specific CTL activities of splenocytes also increased great ly. FACS analysis showed that the CD3+,　CD4+,　CD8+ T cells and CD28 posi tive cells in draining lymph nodes of MIF-FBL3 group mice increased more marked ly than that of control groups. When the wild type FBL3 cells were injected into the mice immunized with MIF gene-modified FBL3 vaccine, the growth of tumors w ere obviously inhibited and the survival rate of the mice was increased. Conclusion: It is suggested that MIF gene-modified tumor vaccine can induce specific antitumor immune responses mediated by T cells and may be a candidate for gene therapy of tumor.

The number and function analysis of T cell from mice immunized with MIF gene-modified tumor vaccine / 第二军医大学学报

Objective: To investigate the role of T cell in the antitumor immune responses induced by MIF gene-modified tumor vaccine. Methods: MIF gene was transferred into FBL3 erythroleukemia cel l by adenovirus carrier and a new type of tumor vaccine was prepared. The chang es of the number and the function of T cell in spleen and lymph node was observe d. Results: After the mice were immunized with MIF gene-m odified FBL3 vaccine, the number of lymphocyte in spleens and lymph nodes increa sed markedly and the specific CTL activities of splenocytes also increased great ly. FACS analysis showed that the CD3+,　CD4+,　CD8+ T cells and CD28 posi tive cells in draining lymph nodes of MIF-FBL3 group mice increased more marked ly than that of control groups. When the wild type FBL3 cells were injected into the mice immunized with MIF gene-modified FBL3 vaccine, the growth of tumors w ere obviously inhibited and the survival rate of the mice was increased. Conclusion: It is suggested that MIF gene-modified tumor vaccine can induce specific antitumor immune responses mediated by T cells and may be a candidate for gene therapy of tumor.

Fetal Bone Marrow Stromal Cells in Cooperation with Exogenous Cytokines Support In Vitro Expansion of Cord Blood CD34(+) Cells / 中国实验血液学杂志

To elucidate the role of fetal bone marrow stromal cells (FBMSC) in cooperation with exogenous cytokines in supporting the in vitro expansion of cord blood CD34(+) cells which were purified by negative immunomagnetic selection, FBMSCs were cultured with different combinations of cytokines including SCF, IL-3, IL-6, FL, G-CSF and EPO in a 14-day liquid culture system. The results showed FBMSC plus SCF, IL-3, IL-6, FL and EPO was the most effective combination which increased total nucleated cells, CFU-GM, BFU-E and CD34(+) cells by (692.4 +/- 52.7) fold, (237.1 +/- 106.6) fold, (114.8 +/- 32.8) fold and (25.3 +/- 10.1) fold, respectively. Our studies indicated that fetal bone marrow stromal cells combined with above-mentioned cytokines can efficiently expand cord blood CD34(+) cells.