Preparation of 4-sulfonylcalix6arene-modified cotton for uranium contamination removal / 中国辐射卫生

Objective To prepare 4-sulfonylcalix[6]arene-modified cotton fibers for adsorption and removal of uranium based on the specific complexation of calix[6]arene with uranium (VI). Methods Chemical grafting was used for the modification of cotton, which reacted with α-bromoisobutyryl bromide, glycidyl methacrylate, and 4-sulfonylcalix[6]arene. Scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), and infrared spectroscopy (FTIR) were used to characterize the structure of 4-sulfonylcalix[6]arene-modified cotton (Cotton S-C[6]a). A Franz diffusion cell was used to simulate uranium-contaminated skin. Laser fluorimetry was used to determine the uranium content. Results SEM, XPS, and FTIR showed that cotton fibers were successfully grafted with 4-sulfonylcalix[6]arene. The optimal conditions of Cotton S-C[6]a for the adsorption of uranium (VI) was pH 4.0, duration of 20 min, and 20 mg of adsorbent. The adsorption process fitted well with pseudo-secondary-order kinetics. The uranium removal efficiency of Cotton S-C[6]a was up to 78.46% in aqueous solution and 81.72% on skin. Conclusion The synthesized Cotton S-C[6]a is highly efficient in the removal of uranium (VI) in solution and on contaminated skin.

Role of complement in radiation-induced lung injury / 中国辐射卫生

@#<b>Objective</b> To investigate the role of complement in radiation-induced lung injury in mice after chest irradiation with ⁶⁰Co γ-rays at a single dose of 20 Gy. <b>Methods</b> C57BL/6 mice underwent chest irradiation with ⁶⁰Co γ-rays at a single dose of 20 Gy, followed by observation for the inflammatory reaction of the lung tissue in the early stage (within 15 d) and pulmonary fibrosis in the later stage (30 and 180 d). Enzyme-linked immunosorbent assay was used to measure the levels of C2, C3a, C4, and C5b-9 in the lung tissues at 1, 3, 7, 15, 30, and 180 d after irradiation. The expression of complement mRNA in BEAS-2B cells after irradiation was determined using RT-PCR. <b>Results</b> Radiation-induced lung injury in micepresented as inflammatory response in the early stage and fibrosis in the late stage. Complement C2, C4, and C5b-9 complexes were increased in the early period (3 or 7 d) after irradiation (<i>P</i> < 0.05), which might be associated with the inflammatory response induced by irradiation. During 3 to 180 d, complement C3a was significantly higher in the irradiated mice than in the control mice, suggesting a close relationship between C3a and radiation-induced lung injury. The irradiated cells showed increased mRNA expression of C2 and C3, with no changes in the mRNA levels of C4 and C5. <b>Conclusion</b> Different complement proteins have varying responses to radiation-induced lung injury, among which C3a is closely related to radiation-induced lung injury, suggesting that regulating C3a and its receptors may be a new way to prevent and treat radiation-induced lung injury.

Antigen presentation and T cell activation by dendritic cells in radiation damage / 中国辐射卫生

@#<b>Objective</b> To explore dendritic cells (DCs)-mediated antigen presentation for radiation-injured cells by using the <i>in vitro</i> cell co-culture technology to simulate the <i>in vivo </i>microenvironment of the lung tissue. <b>Methods</b> ⁶⁰Co γ-irradiated mouse lung epithelial cells (MLE-12) were cultured with bone marrow-derived DCs and/or splenic T lymphocytes for 48 hours. Flow cytometry was used to measure the expression levels of costimulatory molecules (CD80/86) and antigenic peptide recognition complexes (the major histocompatibility complex [MHC] class Ⅰ/Ⅱ) on DCs and T cell activation markers (CD69/28/152) as well as the numbers of CD4⁺ and CD8⁺ T cells. <b>Results</b> ⁶⁰Co γ irradiation significantly increased the apoptosis rate of MLE-12 cells in a dose-dependent manner, and significantly stimulated the expression of CD80/86 and MHC Ⅱ on DCs, without direct activation of T cells. After γ (6 Gy)-irradiated MLE-12 cells were co-cultured with DCs and T lymphocytes for 48 h, there were significant increases in the expression of CD69 and CD28 on T cells, the numbers of CD4⁺ and CD8⁺ T cells, and the expression of CD86 and MHC I on DCs, as compared with the control groups. <b>Conclusion</b> Radiation-injured cells can stimulate antigen presentation by DCs and activate T cells.

Study of sensitization effect of hydroxyurea on chemoradiotherapy of human glioma U251 cells / 中华放射肿瘤学杂志

Objective:To investigate the effect of hydroxyurea (HU) combined with temozolomide (TMZ) and radiotherapy (RT) on the sensitivity of human glioma U251 cells to chemoradiotherapy (CRT).Methods:Human glioma U251 cell line was cultured in vitro. CCK8 cell assay was used to detect the proliferation activity of U251 cells treated with different concentrations of HU/TMZ under different conditions. Flow cytometry was utilized to detect apoptosis rate and cell cycle distribution of U251 cells. Transwell chamber assay and scratch test were performed to evaluate the changes of cell invasion and migration. The expression levels of apoptosis proteins were determined by Western blot. Colony formation assay was adopted to detect the cell survival fraction . Results:HU concentration at 50μmol/L and below did not affect the proliferation of human glioma U251 cells ( P>0.05). Low-dose HU combined with CRT significantly inhibited cell proliferation ( P<0.05), invasion ( P<0.01) and migration (12h P<0.001, 24h P<0.01), and promoted cell apoptosis ( P<0.01) compared with the use of CRT alone. Application of 50μmol/L HU combined with RT increased the radiosensitivity of cells (SER=1.49), significantly prolonged the cell cycle of S phase and G 2 phase (both P<0.05), considerably up-regulated the expression levels of the apoptosis-associated proteins of Caspase-3 and Bax and significantly down-regulated the expression level of anti-apoptosis protein of Bcl-2(all P<0.001). Conclusions:Compared with CRT, HU combined with CRT can further inhibit the proliferation, invasion and migration of human glioma U251 cells, promote cell apoptosis, increase the radiosensitivity and prolong the cell cycle of S and G 2 phases, thereby enhancing the sensitivity of human glioma U251 cells to CRT.

Preliminary mechanism study on the X-linked parkinsonism caused by RAB39B gene mutation c.536dupA / 中华神经科杂志

Objective:To detect the expression level of RAB39B gene and the effect of RAB39B on autophagy and α-synuclein, and then investigate the role of RAB39B gene mutation c.536dupA in the pathogenesis of Parkinson′s disease.Methods:Based on the novel RAB39B gene c.536 dupamutation identified in the previous work, the recombinant expression plasmid (pcDNA3.1-HA-RAB39B-536) of RAB39B gene with this mutation and wild-type recombinant expression plasmid (pcDNA3.1-HA-RAB39B) of RAB39B gene were constructed, and the recombinant expression plasmid was transfected into N2a cells with liposome as experimental group. The control group was made up with N2a cells transfected with plasmid pcDNA3.1-HA-RAB39B. Real-time polymerase chain reaction, Western blotting, immunofluorescence and immunoprecipitation techniques were used to detect the expression level of mutant RAB39B gene and the effects of RAB39B on autophagy and α-synuclein.Results:In the N2a cell model, the transcription level of mutant RAB39B was about twice that of wild type RAB39B, while the protein level of mutant RAB39B (0.30±0.00) was significantly lower than that of wild type (1.50±0.25, t=8.313, P<0.05). After adding proteasome inhibitor MG132, the protein level of mutant RAB39B increased (0.70±0.10, t=6.925, P<0.05); the level of microtubule-associated protein 1 light chain 3 BⅡ/Ⅰ of mutant RAB39B (3.11±0.30) was significantly lower than that of wild type (7.03±0.20, t=18.831, P<0.05); overexpression of wild type and mutant RAB39B did not affect the level of endogenous α-synuclein; overexpression of wild-type RAB39B resulted in elevated level of exogenous wild-type (p.A53T; from 0.60±0.11 to 1.25±0.08, t=8.254, P<0.05) and mutant (from 0.55±0.08 to 1.15±0.08, t=9.293, P<0.05) α-synuclein. Conclusions:The stability of the RAB39B protein decreased with the appearance of c.536 dupA mutation, the mutant protein may be degraded through the ubiquitin-proteasome pathway, and this mutation may affect the autophagy level of cells. RAB39B protein may interact with α-synuclein in vivo and may be involved in the maintenance of the stable level of α-synuclein.

Distribution characteristics and risk factors of infectious pathogens after open limb fracture surgery / 中国基层医药

Objective To investigate the distribution characteristics and risk factors of infectious pathogens after open limb fracture surgery.Methods From January 2016 to December 2018,180 patients with open limb fracture admitted to Hangzhou Hospital of Zhejiang Medical and Health Group were selected to observe the infection after operation.Pathogens were isolated and identified by automatic biological analyzer and bacterial identification system,and drug resistance test was carried out by K-B method.Multivariate logistic regression analysis was used to analyze the risk factors of infection after open limb fracture surgery.Results Among 180 cases of open limb fracture,29 cases had postoperative infection,the infection rate was 16.11％.Among 29 cases of post-operative infection,34 strains of pathogenic bacteria were isolated,including 19 strains of Gram-positive bacteria,13 strains of Gram-negative bacteria and 2 strains of fungi.The resistance rate of Staphylococcus aureus to penicillin G was high,and the resistance rate was 80.00％.Univariate analysis showed that there were no statistically significant differences in gender,BMI,injury site,smoking history,hypertension and intraoperative bleeding between the two groups (x2 =0.252,0.416,0.734,0.856,0.572,all P ＞ 0.05).There were statistically significant differences in age,diabetes mellitus,operation time,hospitalization time,wound contamination and drainage tube placement time between the two groups (x2 =21.537,9.664,17.244,15.459,24.327,19.804,all P ＜ 0.05).The single factor analysis showed that age ＞ 60 years old,diabetes mellitus,operation time ＞ 3h,hospitalization time ＞ 14d,wound contamination and drainage tube placement time ＞ 4d were independent risk factors for infection after open limb fracture surgery.Conclusion Postoperative infection rate of open limb fracture is high.Gram-positive bacteria are the main pathogenic bacteria.Postoperative infection is affected by many factors.In order to prevent postoperative infection,specific measures should be taken and antibiotics should be used reasonably in strict accordance with drug sensitivity test.

Expression of programmed death ligand 1 in liver cancer and its clinical significance / 肿瘤研究与临床

Objective@#To investigate the expression of programmed death ligand 1 (PD-L1) in liver cancer tissues and its clinical significance.@*Methods@#The expression levels of PD-L1 in 110 liver cancer tissues, including 95 cases of hepatocellular carcinoma and 15 cases of cholangiocarcinoma were detected by using immunohistochemical staining method, and the relationship between PD-L1 expression and the clinicopathological characteristics of patients with hepatocellular carcinoma was analyzed.@*Results@#Immunohistochemistry results showed that the positive rate of PD-L1 in liver cancer tissues was 69.1% (76/110), and the positive rate of membrane and cytoplasm was 46.4% (51/110) and 22.7% (25/110), respectively. The positive rate of PD-L1 expression in hepatocellular carcinoma was higher than that in cholangiocarcinoma [78.9% (75/95) vs. 6.7% (1/15)], and the difference was statistically significant (χ 2 = 31.693, P < 0.01). The positive expression of PD-L1 was closely associated with the depth of invasion and TNM stage of hepatocellular carcinoma (both χ2 = 4.629, both P = 0.031), but there was no relationship with the patients'age, gender and pathological grade (all P > 0.05). Further analysis showed that protein expression localization of PD-L1 was closely related to the pathological grade in hepatocellular carcinoma (P = 0.013).@*Conclusion@#The positive expression of PD-L1 in hepatocellular carcinoma is closely associated with depth of invasion and TNM stage, which provides a theoretical basis for the immunotherapy of liver cancer targeting PD-L1.

Expression of programmed death ligand 1 in liver cancer and its clinical significance / 肿瘤研究与临床

Objective To investigate the expression of programmed death ligand 1 (PD-L1) in liver cancer tissues and its clinical significance. Methods The expression levels of PD-L1 in 110 liver cancer tissues, including 95 cases of hepatocellular carcinoma and 15 cases of cholangiocarcinoma were detected by using immunohistochemical staining method, and the relationship between PD-L1 expression and the clinicopathological characteristics of patients with hepatocellular carcinoma was analyzed. Results Immunohistochemistry results showed that the positive rate of PD-L1 in liver cancer tissues was 69.1％(76/110), and the positive rate of membrane and cytoplasm was 46.4％(51/110) and 22.7％(25/110), respectively. The positive rate of PD-L1 expression in hepatocellular carcinoma was higher than that in cholangiocarcinoma [78.9％ (75/95) vs. 6.7％ (1/15)], and the difference was statistically significant (χ2= 31.693, P< 0.01). The positive expression of PD-L1 was closely associated with the depth of invasion and TNM stage of hepatocellular carcinoma (bothχ2= 4.629, both P= 0.031), but there was no relationship with the patients' age, gender and pathological grade (all P> 0.05). Further analysis showed that protein expression localization of PD-L1 was closely related to the pathological grade in hepatocellular carcinoma (P=0.013). Conclusion The positive expression of PD-L1 in hepatocellular carcinoma is closely associated with depth of invasion and TNM stage, which provides a theoretical basis for the immunotherapy of liver cancer targeting PD-L1.

Identification of monoclonal antibodies against hepatoma stem cells by screening for hepatoma spheroid cells / 实用肿瘤学杂志

Objective The objectives of this study were to screen and identify monoclonal antibodies against hepatoma stem cells by screening for hepatoma spheroid cells,and to provide candidate therapeutic monoclonal antibodies for targeting cancer stem cells to treat hepatic cancer. Methods Hepatic cancer stem cells were enriched by serum-free suspension culture. Immunofluores-cence,cisplatin resistance assay, Real -time qPCR, subcutaneous tumor formation in nude mice, and other methods were used to screen and identify anti-hepatocarcinoma stem cell monoclonal antibodies. Immunohistochemistry was used to identify the expression of antigen recognized by monoclonal antibody in liver cancer tissues. The antigen was identified by mass spectrometry. Results MH-CC97L cells were able to form cell spheres in serum -free suspension culture and were labeled with PKH26 dye. Flow cytometry showed that the expression of CD90 + in MHCC97L spheroid cells was 3. 4 times higher than that in the parental cells. In the inhibition experiment of serum-free spheroid,6 monoclonal antibodies significantly inhibited MHCC97L cells in serum-free medium,and in-hibitory rates were 54. 67% ,50. 33% ,45. 73% ,42. 26% ,39. 11% ,and 37. 63% ,respectively. The results of immunofluorescence showed that monoclonal antibodies 28C10 and CD90 were colocalized in MHCC97L cells. The results of real-time qPCR showed that the expression of Sox-2 and Oct-4 in MHCC97L 28C10 + cells was significantly higher than those of MHCC97L 28C10 - cells. Flow cytometry showed that the ratio of 28C10 + in MHCC97L cells and its sphere cells were 7. 98% and 10. 7% ,respectively. The ratio of 28C10 + cells was increased by 1. 34 times. The in vitro globing ability and invasive ability of 28C10 + cells obtained by flow cytometry were significantly higher than those of 28C10 - cells. The results of CCK-8 assay showed that 28C10 + cells were resistance to cispla-tin in 28C10 - cells,which are 1. 96 g/ml and 1. 16 g/ml,respectively. Tumorigenic assay showed that 28C10 + cells were inoculated subcutaneously with 2×104 cells into the nude mice,and tumors were formed in 2 months,with 40% of tumor formation rate. Another nude mouse that did not form a tumor had formed a lung metastasis(1/5). Immunohistochemistry showed that the target antigen posi-tive rate of monoclonal antibody 28C10 in hepatic cancer tissues was about 72. 0% (77/107),while it was lowly expressed in adjacent tissues,and the difference was significant. Mass spectrometry showed that the antigen recognized by 28C10 was HSP90α. Conclusion The MHCC97L spheroid cell model is successfully used to identify a monoclonal antibody that specifically recognizes hepatoma stem cells,which provides a foundation for antibody therapy targeting hepatic cancer stem cells.

Network Pharmacology Exploration of the Mechanism of Cistanche deserticola in the Treatment of Osteoporosis / 中国药房

OBJECTIVE： To study the mechanism of Cistanche deserticola in the treatment of osteoporosis by network pharmacology. METHODS： The active components of C. deserticola were retrieved and obtained by TCM system platform （TCMSP）. Reverse molecular docking server DRAR-CPI and related databases GeneCards and OMIM were used to screen the target of C. deserticola active ingredients in the treatment of osteoporosis. The “component-target”network of C. deserticola was constructed by Cytoscape software， and the interaction between targets was plotted by String database and Cytoscape software. The combination activity of target and active ingredient was evaluated via molecular docking with Systems Dock WebSite server. GO classification and enrichment analysis and KEGG pathway enrichment analysis were conducted for target genes using DAVID database. RESULTS： Totally 13 active ingredients were screened out from C. deserticola， such as verbascoside， leonurine， geniposidic acid. There were 43 active ingredient-treated potential targets， such as RUNX2， VEGF， IL-6， BGP， TNF. Multiple signaling pathways were involved in target action， such as WNT （Wingless/Integrated）， VEGF， TNF. CONCLUSIONS： This study preliminarily explores and validates the main targets and pathways of C. deserticola in the treatment of osteoporosis， which lay the foundation for further study of its mechanism.

Interfering with glutathione S-transferase P1 induces lung cell damage under irradiation conditions / 中华放射肿瘤学杂志

Objective To investigate the association and mechanism between glutathione S-transferase P1(GSTP1) and radiation-induced lung injury.Methods Two effective GSTP1 siRNAs were designed and synthesized.The normal lung epithelial cell line BEAS-2B cells were transfected with GSTP1 siRNA (experimental group,siRNA-1,siRNA-2) and negative control siRNA (negative control group,NC).Western blot was performed to detect the expression levels of GSTP1 protein and EMT-related proteins.CDNB was adopted to evaluate the activity of GSTs.DCFH-DA probe was used for incubation.Flow cytometry was conducted to detect the median fluorescence intensity (MFI) and cellular apoptosis.Annexin-v/PI staining was utilized for incubation.MTT assay was performed to measure the proliferation of BEAS-2B,and the growth curve was drawn based on the results.Results After radiation,compared with the NC group,the ROS level and MFI were significantly higher in experimental group (6774.66±399.60 vs.8759.00±256.96 vs.9967.67±735.11,P＜0.05).In the experimental group,the percentage of cellular apoptosis was remarkably higher than that in the NC group (12.3± 1.16 vs.17.38± 1.65 vs.22.88± 1.20,P＜0.05).MTT assay demonstrated that the OD values in the experimental group were significantly lower than that in the NC group everyday.Further more,the level of EMT process is higher in the experimental group.Conclusions Interfering with the GSTP1 expression in lung epithelial cells can increase the intracellular ROS level,increase the percentage of cellular apoptosis,and reduce the cell proliferation rate following γ-radiation.Besides,it can also promote the epithelial mesenchymal transition in lung epithelial cells.The down-regulation of GSTP1 protein expression level probably aggravates the radiationinduced lung cell injury and promotes the epithelial mesenchymal transition.

Roles of PIF1 helicase in cell cycle arrest induced by ionizing radiation / 军事医学

Objective To observe the effect of PIF1 knockdown on cell growth and cell cycle arrest induced by ionizing radiation.Methods HeLa cell lines that consistently down-regulated PIF1 were prepared by the lentivirus granules interfering technology and confirmed by real-time PCR and Western blotting.The effect of down-regulation of PIF1 on cell growth and cell cycle arrest induced by ionizing radiation was evaluated by cell counting and flow cytometry.Results HeLa cell lines consistently down-regulating PIF1 were established.The growth of HeLa that down-regulated PIF1 was inhibited greatly after 4 Gy of γ-ray irradiation.There was little cell proliferation until the 5th day post 4 Gy γ-ray.Moreover, the S phase block and G2/M phase block of PIF1 knock-downed cell lines were significantly delayed after 8 Gy γ-ray irradiation.Conclusion Knockdown of PIF1 can significantly enhance the radiation sensitivity and delayes the S phase block and G 2 /M phase block induced by ionizing radiation.

Role of Hes1 in malignant transformation of human bronchial epithelial cells induced by tobacco / 中国病理生理杂志

AIM: To investigate the role of transcription factor hairy and enhancer of split 1 (Hes1) in the malignant transformation of human bronchial epithelial cell line BEP2D induced by tobacco.METHODS: The BEP2D cells were chronically exposed to cigarette smoke condensate (CSC) at 1 cigarette per L until the 70th generation.The phenotype of malignant transformation of the cells induced by CSC was detected by soft agar clony formation assay.RT-PCR and Western blot were used to determined the expression of Hes1 at mRNA and protein levels in each generation of the cells.The proliferation and apoptosis of the BEP2D cells exposed to CSC were analyzed with the methods of MTT assay, flow cytometry and cell colony formation assay after treatment with Notch pathway bloker DAPT or liposome transfection with Hes1-siRNA.The expression of Hes1 in the peripheral small airway tissues of the smoking rats was evaluated by immunohistochemical staining.The expression of Hes1 in non-small-cell lung cancer and normal airway tissues was also detected by the methods of immunohistochemistry and RT-PCR.RESULTS: The BEP2D cells in the 70th generation had a malignant transformation phenotype.The expression of Hes1 in the BEP2D cells exposed to CSC for different time showed an increa-sing trend.DAPT and liposome transfection with Hes1-siRNA down-regulated the expression of Hes1, inhibited the cell proliferation and induced cell apoptosis.The expression of Hes1 in the airway mucosa of the rats exposed to cigarette smoke for 1 month and 6 months was significantly higher than that in control group.Cigarette smoking induced the expression of Hes1 in lung cancer and normal airway tissues.CONCLUSION: Hes1 may be involved in smoking-induced lung cancer by promoting the imbalance between apoptosis and proliferation.

Enhancement of Gastric Ulcer Healing and Angiogenesis by Hepatocyte Growth Factor Gene Mediated by Attenuated Salmonella in Rats

The present study developed an oral hepatocyte growth factor (HGF) gene therapy strategy for gastric ulcers treatment. An attenuated Salmonella typhimurium that stably expressed high HGF (named as TPH) was constructed, and the antiulcerogenic effect of TPH was evaluated in a rat model of gastric ulcers that created by acetic acid subserosal injection. From day 5 after injection, TPH (1 × 10⁹ cfu), vehicle (TP, 1 × 10⁹ cfu), or sodium bicarbonate (model control) was administered orally every alternate day for three times. Then ulcer size was measured at day 21 after ulcer induction. The ulcer area in TPH-treated group was 10.56 ± 3.30 mm², which was smaller when compared with those in the TP-treated and model control groups (43.47 ± 4.18 and 56.25 ± 6.38 mm², respectively). A higher level of reepithelialization was found in TPH-treated group and the crawling length of gastric epithelial cells was significantly longer than in the other two groups (P < 0.05). The microvessel density in the ulcer granulation tissues of the TPH-treated rats was 39.9 vessels/mm², which was greater than in the TP-treated and model control rats, with a significant statistical difference. These results suggest that TPH treatment significantly accelerates the healing of gastric ulcers via stimulating proliferation of gastric epithelial cells and enhancing angiogenesis on gastric ulcer site.

Construction of rat models of acute high altitude stress-induced gastrointestinal injury / 中国组织工程研究

BACKGROUND:At present, there are few reports about construction of the model of acute high altitude stress-induced gastrointestinal injury. Construction of a stable high altitude stress-induced gastrointestinal injury model has become a premise of the current studies on acute stress-induced gastrointestinal injury and related diseases. OBJECTIVE: To establish rat models of acute high altitude stress-induced gastrointestinal ischemia/reperfusion injury. METHODS:Twenty-four male Wistar rats were randomly divided into normal control (lower limit of high altitude) , sham (high altitude) and ischemia/reperfusion (high altitude) groups (n=8 rats/group). Rats from al groups underwent a rush for high altitude region. The root of the superior mesenteric artery of rats in the ischemia/reperfusion group was clamped by a vascular clip to completely block blood flow for 60 minutes, and then intestinal blood flow was recovered for reperfusion for 60 minutes. The rats in the sham group were only 

Effects of moxa smoke with different concentrations on expression of SOD and MDA in lung and serum of rats / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of intervention of moxa smoke with different concentrations on superoxide dismutase (SOD) and malondialdehyde (MDA) in serum and lung of male rats, so as to explore the safety concentration of moxa smoke.</p><p><b>METHODS</b>A total of 32 Wistar male rats were randomly divided into a control group, a low-concentration group, a moderate-concentration group and a high-concentration group, 8 rats in each one. All the rats were exposed in the full-automatic toxicant exposure cabinet, and the overshadow of moxa smoke was set at 0%, 10%, 40% and 70%, respectively. Each rat was exposed for 20 min per day. After 26 weeks, the activities of SOD and content of MDA in serum, lung organ and bronchoalveolar lavage fluid were tested.</p><p><b>RESULTS</b>Compared with the control group, the activities of serum SOD in the high-concentration group were reduced (P< 0. 05), but those in the low-concentration group and moderate-concentration group were not significantly different (both P>0. 05). Compared with the control group, the content of serum MDA in the low-concentration group, moderate-concentration group and high-concentration group was increased insignificantly (all P>0. 05). There were no significant differences regarding activities of SOD and content of MDA in lung organ and bronchoalveolar lavage fluid among each moxa smoke group (all P>0. 05).</p><p><b>CONCLUSION</b>There is no obvious toxic reaction in the low-concentration group and moderate-concentration group; in the high-concentration group the antioxidant ability is damaged due to long-term exposure.</p>

Clearance of insoluble depleted uranium particles in lungs by citric acid and ambroxol / 军事医学

Objective To investigate the effect of citric acid and ambroxol on clearing insoluble particles of depleted uranium in rat lungs by establishing a tracheal perfusion model.Methods One hundred and fifty male Wistar rats were randomly divided into model exposure group, normal control group(NC group), depleted uranium exposure group(DU), citric acid treatment group( CA) , ambroxol treatment group( AM) and citric acid+ambroxol treatment group( CA+AM) . The rats were sacrificed on 7, 15 and 30 days.Uranium content in the lungs was detected by microwave digestion method, pathological changes in the lungs were observed, and inflammatory factors of lung homogenates were detected.Results Compared to DU control group, the intrapulmonary uranium deposit amount in experimental groups was significantly reduced on 7 and 15 days (P<0.05).HE stained lung tissue showed that the pathological changes in treatment groups were less significant than in DU control group.The level of IL-1α,IL-1β,and IL-2 was significantly lower than in DU control, but the level of MCP-1 and MIP-1 was observably higher.Conclusion Citric acid and ambroxol can evidently improve the clear-ance of lung uranium and reduce damnification of lung tissues.Drug treatment can reduce the level of pulmonary inflamma-tory cytokines alleviate the chronic inflammation in the lungs, and enhance the capacity of macrophage to recruitment.

Identification and function of a monoclonal antibody against pancreatic cancer stem cell / 肿瘤研究与临床

Objective To identify-and study a monoclonal antibody (McAb) against pancreatic cancer stem cell in vitro,as well as to provide candidate antibody-drug for cancer stem cell-targeted therapy of pancreatic cancer.Methods Cell culture in serum-free medium and PKH26 staining were used to determine the existence of cancer stem cell in PANC-1 cell line.Flow cytometry was used to detect the expression of CD24 and CD44 in PANC-1 cells and sphere cells,Immunofluorescence was used to detect the expression of CD24 and antigen recognized by 15D2.The effects of 15D2 on self-renewal,proliferation and chemosensitivity to gemcitabine of PANC-1 parent or sphere cells were identified by serum-free suspension culture and CCK-8 assay,Immunohistochemistry was applied to detect the level of antigen recognized by 15D2 in cancer and adjacent tissues.Results PANC-1 cells could survive,proliferate and form sphere cells in serum-free medium.The sphere-forming rate was (2.5±0.5) ％.The percentage of CD44+ CD24+ cells population in sphere cells increased by 11.4 folds compared to PANC-1 cells,in which single nearly 97 ％ CD24+ cells was CD44+ CD24+ cells.Therefore,CD24+ was selected for cancer stem cell marker in PANC-1 in this study.The two-color immunofluorescence assay showed that 15D2 could recognize cells which was also stained by CD24.In vitro functional experiments demonstrated that 15D2 significantly suppressed the sphere formation of PANC-1 cells,with the inhibitory rate being 30.4 ％.Meanwhile,the combination of 15D2 and gemcitabine can significantly attenuate the growth of PANC-1 sphere cells.The IC50 was 0.10 μmol/L in 15D2+gemeitabine group,and 0.39 μmol/L in mlgM+gemcitabine group,Immunohistochemical results showed that the antigen recognized by 15D2 was greatly expressed in about 76.9 ％ (11/13) human pancreatic cancer tissues and hardly detected in adjacent normal tissues (10.0 ％,1/10).Conclusion McAb 15D2 can inhibit self-renewal and drug-resistance of pancreatic cancer stem cell in PANC-1 cell line,and it might become a candidate drug for target pancreatic cancer stem cell treatment.

Construction of recombinant adenovirus vector pAdxsi-GFP-HIF containing hypoxia inducible factor gene and its expression in endothelial cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus (pAdxsi-GFP-HIF) encoding human hypoxia inducible factor 1 α gene (HIF-1 α) and to express it in endothelial cells.</p><p><b>METHODS</b>HIF-1 α gene was obtained from human lung cancer cell line A549, which was cultured in hypoxia condition, by RT-PCR. The HIF-1 α gene was subcloned into shuttle vector p Shuttle-CMV-EGFP at KpnI and BamHI sites. After identified with restriction enzymes, plasmid p Shuttle-GFP-HIF was linearized by digestion with restriction endonuclease I-CeuI and I-SceI, and subsequently cotransformed into E.coli DH5a with adenoviral backbone plasmid pAdxsi to make homologous recombination. After linearized by PacI, the homologous recombinant adenovirus plasmid was transfected into 293 cells to package and amplify. The recombinant adenovirus was infected with human umbilical vein endothelial cells (ECV304), and the expression level of HIF-1 α protein was evaluated by ELISA.</p><p><b>RESULTS</b>The recombinant adenovirus vector containing HIF-1 α gene (pAdxsi-GFP-HIF) was successfully constructed and amplified with titer of 3.38 X 10(10) pfu/mL. The green fluorescence protein was detected under fluorescent microscope in ECV304 at 24h after transfection and with a stronger degree after 48h. The concentration of HIF-1 protein was (48.93 ±3.86)ng/mL in supernatant at 48 h after transfection.</p><p><b>CONCLUSION</b>A recombinant adenovirus vector pAdxsi-GFP-HIF, encoding human hypoxia inducible factor 1 α gene, has been constructed in vitro and expressed successfully in ECV304 cells.</p>

Dynamic expression of Nogo-A receptor in the hippocampus following focal cerebral ischemia in rats / 国际脑血管病杂志

Objective To investigate the dynamic expression of Nogo-A receptor (NgR) in the hippocampus following focal cerebral ischemia in rats.Methods The focal cerebral ischemia model was established by electrocoagulating the right middle cerebral artery in rats.The expression of NgR in the ischemic hippocampal CA1,CA2 and CA3 regions at different time points after cerebral ischemia were detected by immunohistochemical staining.Results The expressions of NgR were up-regulated in hippocampal CA1 and CA2 regions at the first day,in the CA3 region at the second day after cerebral ischemia; the expressions of NgR in the CA1 and CA2 regions at the fifth day was decreased to the lowest.The expressions of NgR was up-regulated again in the CA1 and CA2 regions at the 28th day.Conclusions The NgR expression in the hippocampus in ischemic side showed different change characteristics at different regions,however,the overall change trend showed a 2 peaks and a valley phenomenon,which indicated that NgR might have different functions at different time periods after cerebral ischemia.