Identification of serological biomarkers for diagnosis of rheumatoid arthritis using a protein array-based approach / 南方医科大学学报

OBJECTIVE@#To study the cytokine patterns in patients with rheumatoid arthritis (RA) and healthy individuals and identify candidate serum biomarkers for clinical diagnosis of RA.@*METHODS@#This study was conducted among 59 patients diagnosed with RA in our hospital from 2015 to 2019 with 46 age- and gender-matched healthy subjects who received regular physical examinations in our hospital as the control group. Serological autoimmune profiles of 5 RA patients and 5 healthy control subjects were obtained from human cytokine microarrays. We selected 4 differentially expressed cytokines (LIMPII, ROBO3, Periostin and IGFBP-4) and 2 soluble cytokine receptors of interest (2B4 and Tie-2) and examined their serum levels using enzyme-linked immunosorbent assay in 54 RA patients and 41 healthy control subjects. Spearman correlation test was performed to assess the correlation of serum cytokine and soluble receptor expression levels with the clinical features including rheumatoid factor (RF), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), disease activity score (DAS28) and health assessment questionnaire (HAQ). Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic capability of these cytokines.@*RESULTS@#We identified 6 dysregulated cytokines and soluble receptors (2B4, LIMPII, Tie-2, ROBO3, periostin and IGFBP-4) in RA patients (P < 0.01). The serum levels of LIMPII, ROBO3 and periostin were significantly correlated with the disease activity indicators including RF (P < 0.001), CRP (P < 0.001), DAS28 (P < 0.001) and HAQ (P < 0.001) in RA patients. Among the 6 candidate cytokines, 2B4 showed the largest area under the curve (AUC) of 0.861 for RA diagnosis (P < 0.001), followed then by LIMPII, ROBO3, periostin, Tie-2 and IGFBP-4.@*CONCLUSION@#Serum levels of LIMPII, ROBO3 and periostin can be indicative of the disease activity of RA, and serum 2B4, LIMPII, periostin, ROBO3, IGFBP-4 and Tie-2 levels may serve as biomarkers for the diagnosis of RA.

Attaching importance to mechanical perfusion of donor liver and promoting high-quality development of liver transplantation / 器官移植

With the increasing quantity of organ donors and the continual expansion of the definition of extended criteria donor (ECD) livers, the quality of donor liver has become a prominent issue affecting the high-quality development of liver transplantation, which is also the study focus in related fields. Resolving the shortage of organs to the maximal extent and promoting the high-quality development of organ transplantation lead the development direction of organ donation and transplantation in China. In recent years, the application of mechanical perfusion (MP) for the perfusion, preservation, evaluation and repair of donor liver has become a hot topic to improve the quality of liver transplantation within the international community. In this article, according to different conditions of the application of ECD livers in liver transplantation at home and abroad in combination with the research progress on MP in the international community and relevant research experience of our center, the feasibility of establishing an organ intensive care unit (ICU) with integrated organ protection techniques was discussed, aiming to promote the high-quality development of organ transplantation in China and further expand the technical connotation of the "Chinese model" of organ donation and transplantation.

A single center experience in prevention and control of infection risk related to liver transplantation during the COVID-19 outbreak / 中华消化外科杂志

Objective:To investigate the clinical evaluation effects of Corona Virus Disease 2019 (COVID-19) risk assessment scale on preoperative and surgical risk of liver transplantation during the COVID-19 outbreak.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 6 liver transplant recipients who were admitted to Southern Theater Command General Hospital of PLA between January 20 and March 27, 2020 were collected. There were 5 males and 1 female, aged from 42.0 to 62.0 years, with a median age of 53.0 years. There were 6 donors including 5 males and 1 female, aged from 24.0 to 60.0 years, with a median age of 41.5 years. All the donor livers were obtained through the China Organ Transplant Response System. Liver transplantation was performed in the fixed negative pressure operating room, and secondary protective measures were adopted for low-risk donors. Classic orthotopic liver transplantation or Piggyback liver transplantation was performed according to the specific situations of the recipients. Medical staffs in the ward were exposed to the secondary protective measures, and the three-grade protective measures were adopted for medical staffs when the liver transplant recipients had fever or suspected infection. Observation indicators: (1) risk assessment of COVID-19 on liver transplant recipients; (2) risk assessment of COVID-19 on medical staffs of liver transplantation; (3) treatment situations of liver transplant recipients; (4) postoperative situations of liver transplant recipients; (5) follow-up of liver transplant recipients; (6) infection of medical staffs of liver transplantation. Follow-up was performed using outpatient examination or telephone interview to detect whether liver transplant recipients had suspected or confirmed COVID-19 infection up to March 2020. Medical staffs who were involved in organ acquisition, transplantation surgery and ward management were followed up to detect whether they had suspected or confirmed COVID-19 infection within 14 days. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were described as M (range). Count data were expressed as absolute numbers. Results:(1) Risk assessment of COVID-19 on liver transplant recipients: all the 6 recipients and their related families were confirmed no contact with suspected COVID-19 patients or travel history in the epidemic area within 14 days. Of the 6 recipients, 1 was diagnosed with fever with body temperature of 38.1 ℃ and was tested negative for chest computer tomography (CT) examination and nucleic acid test for COVID-19; 1 was diagnosed with fever and hypoxemia with body temperature of 38.5 ℃ and was tested negative for nucleic acid test for COVID-19, and the results of chest CT examination showed large amount of pleural effusion in both lungs without invasive pneumonia; other 4 recipients had no clinical symptoms of COVID-19 with negative results of chest CT examination and nucleic acid test for COVID-19. Five of the 6 recipients had no history of contact with COVID-19 patients and 1 recipient had treatment history at hospital of risk level 1. The preoperative risk level of COVID-19 was low in all the 6 liver transplant recipients. (2) Risk assessment of COVID-19 on medical staffs of liver transplantation: of the 6 recipients, 5 had the waiting hospital of risk level 0 and 1 had the waiting hospital of risk level 1. Six recipients had the transplant hospital of risk level 0. (3) Treatment situations of liver transplant recipients: of the 6 recipients, 2 underwent classic orthotopic liver transplantation and 4 underwent piggyback liver transplantation. The cold ischemia time of liver, time of anhepatic phase, volume of intraoperative blood loss, operation time, treatment time at intensive care unit of the 6 recipients were (5.9±2.4)hours, (49±14)minutes, 1 500 mL(range, 800-1 800 mL), (8.9±2.1)hours, 2 days(range, 1-4 days), respectively. Of the 6 recipients, 2 required adjustment of the immunosuppression program, and 4 did not change the immunosuppression program. (4) Postoperative situations of liver transplant recipients: of the 6 recipients, 5 had no postoperative serious infection and 1 had postoperative serious infection. The 5 recipients without postoperative serious infection had the range of the highest temperature as 37.8-38.5 ℃, and returned to normal temperature within postoperative 3 days. All of the 5 recipients who had no postoperative serious infection received chest CT examination with no obvious manifestation of viral pneumonia and were tested negative for nucleic acid test for COVID-19 at 1 week postoperatively, and then were discharged from hospital. One recipient who had postoperative serious infection had gastrointestinal fistula and repeated fever at postoperative 7 days with the highest temperature as 39.2 ℃. This recipient had body temperature returned to normal and good function of the graft after treatment in the isolation ward with active drainage, and was transferred back to local hospital for further rehabilitation treatment. The duration of hospital stay of the 6 recipients were 30 days(range, 15-74 days). (5) Follow-up of liver transplant recipients: all the 6 recipients were followed up for 31.5 days(range, 12.0-64.0 days) with the normal body temperature, and they had negative results of viral pneumonia for chest CT examination and nucleic acid test for COVID-19. (6) Infection of medical staffs of liver transplantation: surgeons, nurses, anesthetists, medical staffs at ICU and medical staffs at liver transplantation center who participated in liver transplantation had good health within postoperative 14 days, without suspected or confirmed cases of COVID-19 infection.Conclusions:The COVID-19 risk assessment scale has good safety for liver transplant recipients during the COVID-19 outbreak. It is suggested that organ transplantation can be carried out in low-risk recipients and cautiously carried out in recipients of uncertain risk, but organ transplantation should not be carried out in high-risk recipients.

Prognosis factors analysis of patients with malignant solitary pulmonary nodules / 北京大学学报(医学版)

OBJECTIVE@#To explore the prognosis factors that influence the postoperative survival rate in patients with malignant solitary pulmonary nodules and to provide a reference for the prognosis risk stratification of early lung cancer patients.@*METHODS@#In this study, we retrospectively reviewed 172 patients who were admitted to Peking University First Hospital from April 2006 to December 2013. All cases were radiologically defined as solitary pulmonary nodule and were pathologically confirmed to be stage Ia non-small cell lung cancer after surgical procedure. The patients' clinical and follow-up data were summarized and analyzed. The relevance between survival time and factors that may affect patients' prognosis was evaluated, which included gender, age, clinical symptoms, smoking history, comorbidity index, tumor biomarkers, nodule type, type of surgery, nodule location, nodule histopathological type, nodule size, histopathological differentiation grade, proliferating cell nuclear antigen Ki-67 expression level and epidermal growth factor receptor (EGFR) gene mutation. Kaplan-Meier survival analysis, Cox univariant and multivariant regression analysis were conducted to evaluate the factors affecting prognosis.@*RESULTS@#The 3-year overall survival rate of the atients with malignant solitary pulmonary nodules was 93.6%, and the 5-year overall survival rate was 89.8%. KaplanMeier survival analysis and Cox univariant regression analysis showed that the overall survival rate of the male patients was significantly lower than that of the female patients. In addition, the elderly patients with histopathology characterized as high Ki-67 proliferation index were also associated with the worse overall survival (P<0.05). Cox multivariant regression analysis demonstrated that age more than 65 years as well as the high Ki-67 expression level were independent risk factors for overall survival in patients with malignant solitary pulmonary nodules (age: P=0.023, HR=3.531, 95%CI 1.190-10.472; Ki-67: P=0.004, HR=1.021, 95%CI 1.007-1.035).@*CONCLUSION@#For patients with malignant solitary pulmonary nodules, with pathological defined as stage Ia non-small cell lung cancer, age, gender and Ki-67 expression levels might be important prognostic factors. Comprehensive consideration of Ki-67 proliferation index and clinical pathological features may help to stratify the prognosis more accurately and guide the selection of appropriate therapeutic strategies, which needs to be verified by multi-center studies.

Effect of Quercetin on Apoptosis of Platelets and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the effects of quercetin on the apoptosis of platelets and to analyze the intrinsic mechanism.@*METHODS@#Firstly, the effects of quecetin on the apoptosis of platelets was detected by flow cytometry. Secondly, Western blot was used to detect the expression of apoptosis-related proteins in the platelets treated with quercetin for 2 and 4 day.@*RESULTS@#By flow cytometry, it was found that the apoptosis of platelets in the quercetin-treated group (2, 4 and 8 μmol/L) was inhibited, the apoptosis rate of platelets in 2, 4 and 8 μmol/L quercetin group was 3.12%±0.32%, 2.89%±0.15% and 2.31%±0.28%, respectively, which were signigicantly lover than that in control group (P＜0.01). With the increase of quecetin concentration, the proportion ratio of platelets significantly decreased in a concentration-dependent manner（r=-0.9985）. Similar results were observed on the 4th day. Western blot showed that the treatment with quercetin (2, 4 and 8 μmol/L) promoted the expression of anti-apoptotic protein BCL-2, inhibited the expression of pro-apoptotic protein BAX, resulting in a significant increase in the ratio of BCL-2/BAX (P＜0.01), thereby inhibiting the apoptosis of platelets. Similar results were observed on the 4th day.@*CONCLUSION@#Quercetin can inhibit platelet apoptosis by increasing the ratio of apoptosis-related protein BCL-2/BAX in a concentration-dependent manner.

Survey on Assistive Devices Utilization for Old Adults Supported by Community-based Homecare in Dongcheng, Beijing / 中国康复理论与实践

Objective:To investigate the awareness, use, price acceptance, satisfaction, feedback on quality problems, compliance and access methods of assistive devices for the old adults in Dongcheng, Beijing. Methods:From April to December, 2017, 166 homecare old adults from six communities of Dongcheng, Beijing accepted assistive devices after adaption, and were taught how to use. They were followed up once half a month for six months to investigate activities of daily living, satisfaction, awareness and expectation for their assistive devices. Results:There were 22% of them never used their assistive devices. For those assistive devices had been used, 62% were mobile products, 48% were homecare products, 44% were visual products, 20% were hearing products, and 4% were other products. The score of Quebec User Evaluation of Satisfaction with Assistive Technology was (4.04±0.46) in total, with 2% abandoning. Practicalness, good quality, multi-function and high performance-cost were the most important factors in mind to buy the assistive products. Common quality problems were infirmness of assembly (43%), rough appearance (27%), damaged parts (15%), hidden danger (13%), unsuitable size (3%), poor reliability (3%) and lack of structural strength (3%). The ways to obtain assistive devices included self-purchase (80%), government adaptation (13%), family and friends giving (6%), and lease (0%). Conclusion:Assistive devices can improve quality of life for the old adults. They prefer to choosing assistive devices of multi-function, cost-effective, for daily living, communication and homecare. Professional fitting, regular visit and maintenance can increase the satisfaction, and reduce abandonment of assistive devices. Assistive devices leasing service may help to promote the access of assistive devices.

Analysis on clinical data in 36 cases of recurrent glioblastoma retreatment / 重庆医学

Objective To study the factors influencing prognosis in the patients with recurrent glioblastoma muhiforme (GBM) and to investigate the effect of retreatemt.Methods The retrospective analysis method was adopted to collect the clinical and follow up data in 36 cases of recurrent GBM retreatment in the neurosurgery department of this hospital from March 2008 to March 2013.The prognosis influencing factors were analyzed.Results The univariate analysis results showed that the gender,resection degree,treatment mode and initial scheme had the influence on the progression free survival(P＜0.05).The resection degree had an impact on the overall survival(P＜0.05).The multivariate analysis results showed that KPS score,resection degree and treatment mode had effect on the progression free survival(P＜0.05).The resection degree had an influence on the overall survival (P＜0.05).Conclusion If the patients with recurrent GBM still hasthe chance of operation whole excision,the re-treatment can reach the effect for relieving the symptoms,improving the quality of life and prolonging the survival period.

Chemical constituents from stems of Clausena emarginata / 中草药

Objective: To investigate the chemical constituents from the stems of Clausena emarginata. Methods: The compounds were isolated by macroporous resin, silica gel, ODS column chromatography, Sephadex LH-20, reversed-phase MPLC, and then purified by preparative HPLC. Their structures were determined by the analysis of ultraviolet spectrum, mass spectrum, and NMR spectrum. Neuroprotective activities of compounds 11 and 12 were initially investigated. Results: Sixteen compounds were isolated from the petroleum ether and acetone fractions of 95% ethanol extract of the stems of Cl. emarginata, and their structures were identified as 1H-Indole-3-carboxaldehyde (1), E-N-benzoiltiramine (2), dehydrodiconiferyl alcohol (3), tortoside A (4), zhebeiresinol (5), evofolin B (6), 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucopyranoside (7), echipuroside A (8), 3-methylcarbazole (9), murrayafoline A (10), clausine Z (11), indizoline (12), clausenaline B (13), mafaicheenamine A (14), dictamnine (15), and honokiol (16). Conclusion: Compounds 1-8 are isolated from the plants of genus Clausena L. for the first time, compounds 1-16 are isolated from this plant for the first time. Compounds 11 and 12 show the neuroprotective activity against rotenone induced PC12 cell damage.

N-methyl-D-aspartic acid receptor 1 (NMDAR1) aggravates secondary inflammatory damage induced by hemin-NLRP3 pathway after intracerebral hemorrhage / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>Inflammation plays a critical role in secondary brain damage after intracerebral hemorrhage (ICH). However, the mechanisms of inflammatory injury following ICH are still unclear, particularly the involvement of NLRP3 inflammasome, which are crucial to sterile inflammatory responses. In this study, we aim to test the hypothesis that NLRP3 signaling pathway takes a vital position in ICH-induced secondary inflammatory damage and detect the role of N-methyl-D-aspartic acid receptor 1 (NMDAR1) in this progress.</p><p><b>METHODS</b>ICH was induced in mice by microinjection of hemin into the striatum. The protein levels of NMDAR1, NMDAR1 phosphorylation, NLRP3 and IL-1b were measured by Western blot. The binding of NMDAR1 to NLRP3 was detected by immunoprecipitation.</p><p><b>RESULTS</b>The expression of NMDAR1, NMDAR1 phosphorylation, NLRP3 and IL-1b were rapidly increased after ICH. Hemin treatment enhanced NMDAR1 expression and NMDAR1 phosphorylation, as well in cultured microglial cells treated by hemin. Hemin up regulated NLRP3 and IL-1b level, which was reversed by MK801 (NMDAR antagonist) in vitro. Hemin also promoted the binding of NMDAR1 to NLRP3.</p><p><b>CONCLUSION</b>Our findings suggest that NMDAR1 plays a pivotal role in hemin-induced NLRP3-mediated inflammatory damage through synergistic activation.</p>

Development and evaluation of Chinese version of the Manchester patient safety framework / 现代临床护理

Objective To develop Chinese version of the Manchester patient safety framework, and evaluate the feasibility of its clinical application. Methods The Manchester patient safety framework (MaPSaF) was transformed into Chinese version by translation and back-translation together with inquiries from experts. The trial study was done firstly among 136 nurses in five pilot wards and followed by five times of focus-oriented group interviews. A semi-structured, in-depth interview was then performed in 12 randomly selected participants and acquired data were analyzed by category analysis. Result Two themes were abstracted: Chinese version of MaPSaF showed a good practicability, effectiveness and operability and it was effective for deepened understanding of patient′s safety culture. Conclusion The Chinese version of MaPSaF can be used for safety culture evaluation during nursing process in China.

Combination analysis of new drug discovery with "Xiaohe Silian" method and traditional Chinese medicine clinical pharmacy / 中国中药杂志

With the kernel of efficacy, "Xiaohe Silian" was a pattern and method for new drug discovery which was constituted with "metabolism-efficacy, toxicity-efficacy, quality-efficacy and structure-efficacy". Its connotation was in keeping with traditional Chinese medicine (TCM) clinical pharmacy. This paper systematically summarized the research method of new drug discovery practice process for TCM. To avoid western drug like in TCM new drug discovery, we carried out combination analysis with TCM clinical pharmacy. The correlation analysis between basic elements of "Xiaohe Silian(n) and TCM clinical pharmacy was studied to guarantee this method could integrate closely with TCM clinic from all angles. Hence, this method aimed to provide a new method for TCM new drug discovery on the basis of TCM clinical pharmacy with insisting on holistic view of multicomponent study, kinetic view of metabolic process when the curative effect occurred and molecular material view of quality control and structure-activity exposition.

Effect of kinetin on immunity and splenic lymphocyte proliferation in vitro in D-galactose-induced aging rats / 生理学报

The purpose of this paper is to study the effect of kinetin (Kn) on immunity and splenic lymphocyte proliferation in vitro of aging rats induced by D-galactose (D-gal). Fifty SD rats were randomly divided into five groups: control group, aging model group, Kn low dose group, Kn middle dose group and Kn high dose group. The aging model group was proposed by napes subcutaneous injection of D-gal (125 mg/kg) for 45 d, and anti-aging groups were intragastrically administered with 5, 10, 20 mg/kg of Kn respectively from day 11. IgG, IgA, IgM contents of serum, the apoptosis percentage, stimulation index (SI) and proliferation index (PI) of splenic lymphocyte in vitro were evaluated. The results showed that the apoptosis percentage of splenic lymphocyte in aging model rats was higher, the serum IgG, IgA and IgM contents, SI and PI were lower than control group. Kn significantly decreased the apoptosis percentage of splenic lymphocyte, while increased the serum IgG, IgA and IgM contents, SI and PI in aging model group. These results suggest that Kn could inhibit the apoptosis, while promote the proliferation of splenic lymphocyte, and then effectively enhance the immune power of the aging rats and slow down the aging process.

Effect of kinetin on ovary and uterus in D-galactose-induced female mouse model of aging / 生理学报

The present study was to investigate the effect of kinetin on ovary and uterus of D-galactose-induced female mouse model of aging. Aging female mice model caused by D-galactose were used as model group, the aging model mice intragastrically administered with kinetin solution (daily 25 mg/kg or 50 mg/kg) were used as kinetin groups, and the mice with solvent as normal group (n = 20). To detect the effects of kinetin, estrous cycle, estradiol content, ovarian and uterine wet weight and organ index, SOD and GSH-Px activities, MDA and total protein contents, as well as the reserve function of ovaries were examined. The results showed that, kinetin-induced changes in two kinetin groups were observed, compared with the model group: (1) the estrous cycle was shortened; (2) serum estradiol content was significantly increased; (3) the wet weights of the ovary and uterus were increased significantly; (4) SOD and GSH-Px activities of ovary and uterus were significantly higher; (5) the MDA contents of the ovary and uterus were reduced significantly; (6) total protein contents of the ovary and uterus were increased significantly; (7) the numbers of mature oocytes in fallopian tubes were increased significantly. The results show that kinetin can protect ovary and uterus against oxidative damage, prevent low estrogen secretion caused by ovarian oxidative damage, shorten the estrous cycle in mice, and eventually maintain ovarian and uterine vitalities.

Application of atomic force microscopy in ultrastructure and biomechanics of cells and biomacromolecules / 医用生物力学

To be the representative fruition resulted from the rapid development in micro-nano theory and technology, atomic force microscopy (AFM) has greatly promoted the expansion of biological research in micro-nano scale, and facilitated the birth and development of micro-nano biology as an important technique in its 25-year evolutional progress. Based on the fundamental principles and detection modes of AFM, as well as the author’s research findings and work experience in this field, the paper reviews the application of AFM in the study on ultrastructure and biomechanical properties of cells and biomacromolecules in the aspects of biological structure and morphology, surface physicochemical characterization and mechanical manipulation of biological macromolecules, and focus on some important scientific and technical problems on AFM in micro-nano biomedical research needed to be improved and solved urgently, with exploratory insights and recommendations for potential users in ultrastructure and biomechanics of cells and biomacromolecules.

The study of changes on NKT cells of experimental autoimmune encephalomyelitis (EAE) mice / 细胞与分子免疫学杂志

AIM: To observe the changes of the number of NKT cells in spleens and livers of induced model of experimental autoimmune encephalomyelitis (EAE), and to study the role NKT cells play in the immunoregulation of EAE. METHODS: C57BL/6 mice were immunized with MOG peptide and received clinical evaluation daily. The mice were sacrificed at the fastigium and the splenic and hepatic lymphocytes were isolated. The changes of NKT cells in normal and EAE C57BL/6 mice were detected by flow cytometry. RESULTS: The percent of NKT cells in lymphocytes of different organs of EAE model were greater decreased than in that of normal mice. The percent of NKT cells in splenic lymphocytes of normal mice was 2.22± 0.14, while that in EAE mice was 1.94±0.07 (P < 0.05). The percent of NKI cells in hepatic lymphocytes of normal mice was 5.52±2.17, while that in EAE mice was 2.67± 1.41 (P < 0.05). CONCLUSION: The proliferation of splenic and hepatic NKT cells in C57BL/6 mice are inhibited in EAE model, which may indicate that the immune function conducted by NKT cell is down regulated in EAE mice.

Effect of vinblastine nanoparticles on antiproliferation in human glioma cell lines BT325 / 中国中药杂志

<p><b>OBJECTIVE</b>To compare antiproliferation effects of vinblastine nanopraticles and vinblastine water solution in human glioma cell lines BT325.</p><p><b>METHOD</b>Vinblastine nanoparticles were prepared by emulsion polymerization process and using dextran as a stabilizing agent. It was characterized by means of morphology, size, drug entrapment efficiency and loading efficiency. Human glioma cell lines BT325 were treated with different concentrations of vinblastine nanoparticles and vinblastine water solution for 48 h, Antiproliferation effect was measured by MTT method. Morphological changes were observed by inverted microscope, transmission electron microscope and scanning electron microscope.</p><p><b>RESULT</b>Mean diameter of VLB-PBCA-NP was about 74.4 nm, and drug entrapment efficiency and loading efficiency was 78.47% and 39.24%, respectively. Cell growth inhibition rate of vinblastine nanoparticles group and vinblastine water solution group in a concentration range (5-5 000 g x L(-1)) for 48 h was 41%, 49%, 73%, 83% and 28%, 33%, 54%, 60% respectively. Entrapment of VLB in NPS may distinctly degrade absorbency as compared to free drugs. Glioma cell BT325 which treated with VLB water solution were initial stage of apoptosis, and apoptosis body were forming. But VLB NPS-treated BT325 cells were intermediate or end stage, and missed structure integrality.</p><p><b>CONCLUSION</b>VLB-PBCA-NP and VLB water solution could inhibit the growth of human glioma cell lines BT325, and VLB nanoparticles have stronger inhibition effect compared with VLB water solution in the same dose. PBCA may be effective as promising carrier for the transport of vinblastine into the glioma cells.</p>

Study on Gambogic Acid-loaded Polylacticacid Nanoparticles / 中国生物工程杂志

Gambogic acid-loaded polylacticacid nanoparticles (GA-PLA-NPs) were prepared by modified emulsification solvent diffusion. The shape of nanoparticles was observed by transmission electron microscope (TEM).The size distribution and mean diameter were measured by laser particle size analyzer. The entrapment efficiency and content of drug loading were determined by Ultraviolet Spectrophotometer after ultracentrifugation. GA-PLA-NPs release behavior in vitro was carried out. The acute toxicity were carried out to study the security of GA-PLA-NPs. The preparation process adapted to the formulation was as follows: the volume ratio of the aqueous and organic was 2∶1(v/v), the surfactant concentration in aqueous was 0.5%,the drug concentration in organic was 0.1%(w/v), GA∶PLA was 1∶4(w/w). The mean diameter was 51.36nm for the nanoparticles prepared by above conditions.The entrapment efficiency and content of drug loading were 98.87 % and 13.3 %. The release behavior of drug in vitro showed an initial burst effect with subsequently a slower rate stage. The LD50 value of GA-PLA-NPs on mouse was 26.3 mg/kg. The results showed that the GA-PLA-NPs were well prepared with stable quality and high dispersion. PLA-NPs might be used as a new carrier for gambogic acid.

Studies on fermentation conditions and purification of mutant human interleukin-2 expressed in Pichia pastoris / 生物工程学报

Interleukin-2 (IL-2) was initially isolated as a T cell growth factor and had been shown to direct the expansion and differentiation of several hematopoietic cell types. Clinical studies using IL-2 in the treatment of AIDS have been encouraging, due to its critical role as a proliferative signal for activated T-lymphocytes. IL-2 has also undergone trials in the treatment of several types of cancer, based on its stimulation of cytotoxic, antitumor cells. Today, human IL-2 is produced completely by genetically engineered method, and it has been proved that genetically engineered recombinant human IL-2 has almost the same function and clinical effect as wild IL-2. In the former study, recombinant human IL-2 usually comes from E. coli, in this paper the mutant IL-2 was successfully expressed and purified in Pichia pastoris for the first time. As a eukaryote, Pichia pastoris has many of the advantages of higher eukaryotic expression systems such as protein processing, protein folding, and posttranslational modification, while being as easy to manipulate as E. coli or Saccharomyces cerevisiae. It is faster, easier, and less expensive to use than other eukaryotic expression systems such as baculovirus or mammalian tissue culture, and generally gives higher expression level. Expression conditions of human mutant interleukin-2(the codon for cysteine-125 of human IL-2 with alanine; the codon for leucine-18 with methionine; the codon for leucine-19 with serine) in the recombinant Pichia pastoris strain were optimized via test of some factors such as the rate of aeration, the inductive duration, the initial pH and the concentration of methanol. The results from tests showed that the most important parameter for efficient expression of interleukin-2 in recombinant Pichia pastoris strain is adequate aeration during methanol induction, and the optimum inductive condition for interleukin-2 expression was: more than 80% aeration, 2 days for induction, the initial pH of 6.0, the final methanol concentration of 1.0%. With this condition, the expressed IL-2 was secreted into fermentation broth and reached a yield of 30%, approximately 200 mg/L. Expressed interleutin-2 (MvIL-2) was isolated and purified by centrifugation, millipore filtration to concentration, Econo-PacS strongly acidic cation exchanger cartridge and molecular sieve chromatography and the yield of MvIL-2 was 27%. MvIL-2 was purified to electrophoretic purity by SDS-PAGE and only one peak being loaded on HPLC. Purified MvIL-2 protein had stimulating activity similar to the wild type of IL-2 as assayed by IL-2-dependent CTLL-2 cells. However, the stability of MvIL-2 was superior than that of IL-2 at different temperatures. The activity of obtained MvIL-2 was 4 - 5 times of the wild type of IL-2, So MvIL-2 had an advantage over wild type of rhIL-2 in storage stability and activity.

Experimental study of cardiac muscle tissue engineering in bioreactor / 中国医学科学院学报

<p><b>OBJECTIVE</b>This study investigates construction of cardiac muscle cell-porous collagen scaffold complex in a bioreactor so as to unveil the possibility of generating 3-dimensional cardiac muscle tissue under the environment that mimics microgravity in vitro.</p><p><b>METHODS</b>1-2-day old neonatal rat cardiac muscle cells were isolated by sequential digestion and pre-plating methods, then seeded onto porous collagen scaffold. The cell-collagen complex was transferred into rotary cell culture system (RCCS) and incubated for 7 days. Cells cultured in 75 ml flasks and constructs cultures on plates served as control. Morphological changes of the cells were observed by light microscope and metabolic rate was recorded. Ultrastructure of the cells growing in porous collagen was observed by transmission electron microscopy. Content of total DNA and protein in the newly-formed tissue were analyzed. H-E and anti-sarcomeric alpha-actin stains were performed in comparison with native cardiac muscle.</p><p><b>RESULTS</b>The isolated cardiac muscle cells adhered to the bottom of the flasks 24 hours after plating and began to beat spontaneously. When incubated for 7 days in RCCS, cell-collagen constructs of form a continuous outer tissue layer containing cells aligned with each other. The cell population in the interior of the construct was less in density than the outer part. Transmission electron microscopy demonstrated that subcellular elements characteristic of cardiac myocytes were in the outermost layer of constructs. A strongly positive stains of anti-sarcomeric alpha-actin suggested presence of cell population of differentiated cardiac myocytes in these constructs. Construct biomass was not significantly different from that in neonatal rat ventricle and approximately 40% of that in adult rat ventricles. Construsts in plates contained a few of cells which were less than those in RCCS. Metabolic activity of cells cultured in RCCS was higher than that in flasks and plates.</p><p><b>CONCLUSIONS</b>Dissociated cardiac muscle cells cultured on 3-dimensional scaffolds in RCCS under favorable conditions can form engineered constructs with structural and functional features resembling those of native cardiac tissue.</p>