The anti VEGF monoclonal antibody conjugated with nanometer particle of 5-FU

AIM:The characteristics of nanometer particles, which were prepared by the conjugation of anti-VEGF monoclonal antibodies and 5-fluorouracil-loaded polylactic acid nanometer particles (5-FU-NPs), were investigated for improving the anticancer activity of 5-FU. METHODS:The method of couple linkage of chemical bonds was used to prepare the 5-FU-NPs with VEGF antibody, then the appearance, distribution of particle diameter, releasing in vitro and the immunological activity were detected. RESULTS:The 5-FU-Ab-NPs appeared as regular globular, the average particle diameter was (202±23)nm. The 5-FU-Ab-NPs possessed the similar delayed release character of 5-FUs. More than 80% of the immmunogical activity were detected in conjugates-retained antibody by the immunological methods and electron microscopy. CONCLUSION:The 5-FU-Ab-NPs possess the similar delayed released activity to 5-FU-NPs and have double activity of immune targeting and delayed releasing, which may increase the local concentration of 5-FU.     

Function of peritoneal mesothelial cells on the microenvironment by administration of endometrial cells

AIM: To determine the function of peritoneal mesothelial cells on the inflammatory microenvironment by administration of endometrial cells,and further define the pathogenesis of endometriosis.METHODS: Homogenous mouse endometrial epithelial and stromal cells were injected into the peritoneal cavities of Swiss Webster mice.After 4,24,and 72 h,a number of endpoints evaluated: protein concentrations of cytokine MCP-1,IL-1 α,IL-6 in peritoneal lavage and gene expressions of MCP-1,IL-1 α,IL-6 in peritoneal mesothelial cells and macrophages.RESULTS: The intraperitoneal administration of endometrial cells increased the protein expressions of cytokines in the peritoneal lavage of the recipient mice,which increased at 4-hour points and subsequently decreased with time.Gene expressions of cytokines in peritoneal mesothelial cells paralleled with the protein quantities in peritoneal lavage.The peak time of gene expression of cytokines in peritoneal macrophages was at the 24-hour point.The endometrial epithelial cells stimulated stronger inflammatory responses in the peritoneal cavity than the endometrial stromal cells.CONCLUSION: The recipient mice have a non-specific inflammatory response to the presence of endometrial cells in the peritoneal cavity.Mesothelial cells may be the targets of early inflammatory stress initiated in the presence of endometrial cells.     

Overexpressed human FAT10 protein induces the apoptosis in the starved HEK293 cells

AIM: To study the effect of human FAT10 on the apopotosis of HEK293 cells using flag-tagged human FAT10 protein.METHODS: The fragment of FAT10 gene was cloned into the pcDNA3-flag vector,which was identified by PCR,enzyme digestion and sequencing.The reconstructed plasmids were transfected into HEK293 cells.The expression of introduced FAT10 in the normal cultured and starved cells was detected respectively by Western blotting.XTT assay and DNA ladder method were used to analyze the effect of FAT10 on the apoptosis of starved HEK293 cells.RESULTS: The reconstructed plasmids were highly expressed in HEK293 cells with different expression mode at the mormal cultured and starved state.The livability of starved FAT10 overexpressed HEK293 cells was significantly lower than that of normal cultured cells.DNA ladder was observed in the starved FAT10 overexpressed cells,but not in the normal cultured cells.CONCLUSION: The eukaryotic expressed plasmids of flag-tagged FAT10 were constructed successfully,and highly expressed in HEK293.Overexpressed FAT10 enhances the apoptosis in the starved HEK293 cells.     

Lowered Antioxidant Status of Red Blood Cells in Post-Parturient Haemoglobinuria of Buffaloes

The antioxidant status of the red blood cells of buffaloes (n = 20) suffering from post-parturient haemoglobinuria (PPH) was assessed by comparing their tocopherol (vitamin E) and reduced glutathione contents with those of red blood cells from apparently healthy buffaloes (n = 20). The red cell tocopherol content of the diseased buffaloes (1.76±0.11 g/ml) was significantly (p<0.01) lower than that of healthy buffaloes (2.45±0.14 g/ml). This may be the first report comparing the concentration of tocopherol in the red blood cells of buffaloes suffering from PPH and apparently healthy buffaloes.There was a drastic reduction in the reduced glutathione content in the red cells of haemoglobinuric buffaloes (23.74±2.86 mg%) compared to the healthy control buffaloes (73.71±3.87 mg%). The diseased buffaloes also exhibited severe hypophosphataemia. These findings suggest that an impaired or insufficient antioxidant potential of the red blood cells in this disease in buffaloes is associated with the phosphorus deficiency.     

27-Hydroxycholesterol modulates lung cancer cell proliferation by activation of LXR signaling pathway

AIM:To investigate the effect of cholesterol metabolite 27-hydroxycholesterol (27-OHC) on the proliferation of lung cancer cells. METHODS:Human lung cancer A549 cells were treated with 27-OHC at different concentrations (0, 0.3125, 0.625, 1.25, 2.5, 5 and 10 μmol/L) for 24~48 h. The cell viability, cell cycle, cell prolife-ration, the intracellular cholesterol levels and cholesterol metabolism-related molecule expression were subsequently assessed by CCK-8 assay, flow cytometry, EdU staining, tissue total cholesterol detection kit, real-time PCR and Western blot. RESULTS:27-OHC decreased the viability of the A549 cells in a dose-and time-dependent manner (P<0.01) and inhibited the cell proliferation (P<0.05). The expression of typical liver X receptor (LXR) downstream target proteins including ATP-binding cassette transporter A1 (ABCA1), low-density lipoprotein receptor (LDLR), and 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CR) were modulated, which promoted the efflux of intracellular cholesterol, and reduced cholesterol influx and de novo synthesis, resulting in decreased intracellular cholesterol levels and cell viability. Furthermore, the inhibitory effect of 27-OHC on A549 cell viability was significantly attenuated after the LXR pathway was partially blocked by 5 μmol/L GSK2033 treatment (P<0.05). CONCLUSION:27-OHC inhibits A549 cell prolife-ration via activation of LXR signaling pathway.     

Effects of Sini decoction on the ultrastructure of small intestinal epithelial cells in rats with intestinal ischemia-reperfusion

AIM: To investigate the effect of Sini decoction (SND)on the ultrastructure of small intestinal epithelial cells in rats with intestinal ischemia-reperfusion. METHODS: Thirty-two Sprague-Dawley rats of both sexes were randomly divided into 4 groups: (1) Control group in which sham operation was performed; (2) Model group in which intestinal I/R was produced by clamping super mesenteric artery(SMA) for 1 hour and declamping SMA for 3 hours; (3) SND1 group in which SD (0.6 g/200 g rat) was given via stomach tube 3 d before intestinal I/R; (4) SND2 group in which SD (1.2 g/200 g rat)was given via stomach tube 3 d before intestinal I/R. A strip of small intestine was taken from distal end of ileum for electron microscopic examination. The two-dimensional structural parameters and three-dimensional structural parameters of mitochondria were calculated. RESULTS: (1)Morphological changes of small intestine: In control group, epithelial cells were orderly arranged, with normal mitochondria and intestinal villi. In model group, the gaps between epithelial cells widened. There were a lot of apoptotic cells. Microvilli were short and swelled. Mitochondria were swelled obviously with broken ridges. Endoplasmatic reticulum was severely dilated. In SND1 and SND2 groups, microvilli and epithelial cells were orderly arranged relatively, mitochondria was slightly swelled. (2) Structural parameters of mitochondria: In model group, there were the least mitochondria and the swelling of mitochondria was severe. In SND1 and SND2 groups, the mitochondria was more than that of model group and the swelling were slight. CONCLUSION: Sini decoction can protect small intestine from ischemia-reperfusion injury without dose-dependent effect.     

High concentration of extracellular ATP causes autophagy and apoptosis of SH-SY5Y cells

AIM：To examine the effects of high concentration of extracellular ATP on human neuroblastoma SH-SY5Y cell injury. METHODS：Cultured SH-SY5Y cells were grouped according to the concentrations of ATP and treatment time. The cell viability was detected by CCK-8 assay. The variation of autophagic vacuoles was observed with monodansylcadaverine staining. The cell apoptosis was analyzed by Hoechst 33258 staining. Meanwhile, apoptotic rate was detected by flow cytometry. The levels of caspase-3 and microtubule-associated protein 1 light chain 3-Ⅱ (LC3-Ⅱ) were determined by Western blotting. RESULTS：Compared with control group, the survival rate of SH-SY5Y cells was significantly reduced by ATP at different concentrations (3, 6, 9, 12 and 15 mmol/L for 3 h) and different treatment time (1, 2, 3 and 6 h with 6 mmol/L ATP, peaking at 3 h). The autophagic vacuoles of SH-SY5Y cells were significantly increased at 1 h with ATP treatment, trended to decrease over time and returned to control level at 6 h. The protein expression of LC3-Ⅱ was significantly increased at 1 h with ATP treatment, which was consistent with the time points of increasing autophagic vacuoles. LC3-Ⅱ expression level gradually decreased at 2~3 h with ATP treatment, and returned to control level at 6 h. Compared with control group, the apoptotic rate and the expression level of caspase-3 were enhanced synchronously. The peak of apoptotic rate occurred at 3 h, and kept until 6 h.The level of cleaved caspase-3 expression peaked at 6 h. CONCLUSION：High concentration of extracellular ATP induces the autophagy and apoptosis of SH-SY5Y cells. The increased autophagy shows up, followed by the climax of apoptosis until 6 h. With the prolonged duration of ATP, apoptosis is the main process in the cells.     

Evaluation of adverse reactions in dogs following intravenous mesenchymal stem cell transplantation

Background

Recent studies have assessed the therapeutic potential and drawbacks of mesenchymal stem cells (MSCs). The adverse reactions of intravenous transplantation of bone marrow (BM)-derived MSCs were examined at varying doses and frequencies of administration.Nine healthy beagle dogs were purchased from a commercial laboratory. The dogs were distributed equally (n = 3 per group) and randomly into three groups. All dogs received allogeneic BM-derived MSCs: 2 × 10⁶ once (group A), 2 × 10⁷ once (group B), and 2 × 10⁶ for three consecutive days (group C). Various laboratory examinations, multi-detector computed tomography features and histopathology were evaluated to clarify the clinical and diagnostic features of adverse reactions of MSCs administration, prior to receiving MSCs (pre procedure) and on days 1, 3, and 7 post transplantation.

Results

Only one dog had clinical signs during and after MSCs transplantation. Dogs receiving 2 × 10⁶ MSCs showed increased numbers of lymphocytes but the total white blood cell counts were not elevated (P < 0.01). Multi-detector computed tomography (MDCT) revealed pulmonary parenchymal changes in one dog and histopathologic examination revealed pulmonary parenchymal edema and hemorrhage in four dogs. The presence of pulmonary thromboembolism was not detected in either examination.

Conclusions

We considered the presence of pulmonary edema and hemorrhage as possible adverse reactions after intravenous MSCs transplantation; however these results should be cautiously interpreted.     

富铁酵母对家兔红细胞免疫功能的影响

为探讨富铁酵母对家兔红细胞免疫功能的影响,将32只家免随机分成富铁酵母高、低剂量组,FeSO4组及空白对照组,分别灌服富铁酵母(Fe元素8 mg/kg、4 mg/kg)、FeSO4(Fe元素4 mg/kg)及空白生理盐水;连续5 d后,检测红细胞C3g受体花环率和复合物(IC)花环率.结果:富铁酵母较FeSO4及空白组能显著提高红细胞C3h受体花环和红细胞复合物(IC)花环的百分率,但高、低剂量间无显著性差异.结论:富铁酵母能显著提高红细胞免疫功能.