Targeted delivery of miRNA-204-5p by PEGylated polymer nanoparticles for colon cancer therapy.

AIM: miRNAs have been recognized for their potential in cancer therapeutics, and multiple miRNAs were suggested to affect target genes expression. To overcome limitations of free synthetic miRNAs, such as easily degraded in biofluids and limited in cellular uptake, novel miRNAs delivery systems need to be developed. MATERIALS & METHODS: Using surface-functionalizing technique, poly(D,L-lactide-co-glycolide)/poly(L-lactide)-block-poly(ethylene glycol)-folate polymer nanoparticle (PLGA/PLA-PEG-FA) loaded with miR-204-5p (FA-NPs-miR-204) was developed. The therapeutic efficacy of FA-NPs-miR-204 was evaluated in the Luc-HT-29 xenograft tumor model in vivo. RESULTS: FA-NPs-miR-204 could be taken up by HT-29  and HCT-116 cells efficiently, resulting in significant inhibitory effect on cell proliferation and promotive effect on cell apoptosis. In vivo study showed that FA-NPs-miR-204 could exert tumor suppressive function in Luc-HT-29 xenograft model. CONCLUSION: Our study demonstrates a convenient miRNA delivery system that targets tumor tissue and exerts tumor suppressive function, thus demonstrating a potential new therapeutic option for colon cancer.

Potential protective effects of Clostridium butyricum on experimental gastric ulcers in mice.

AIM: To investigate the effects of Clostridium butyricum (C. butyricum) on experimental gastric ulcers (GUs) induced by alcohol, restraint cold stress, or pyloric ligation in mice, respectively. METHODS: One hundred and twenty mice were randomly allocated into three types of gastric ulcer models (n = 40 each), induced by alcohol, restraint cold stress, or pyloric ligation. In each GU model, 40 mice were allocated into four groups (n = 10 each): the sham control group; model group (GU induction without pretreatment); C. butyricum group (GU induction with C. butyricum pretreatment); and Omeprazole group (GU induction with Omeprazole pretreatment). The effects of C. butyricum were evaluated by examining the histological changes in the gastric mucosal erosion area, the activities of superoxide dismutase (SOD) and catalase (CAT), the level of malondialdehyde (MDA), and the contents of interleukin (IL)-1ß, tumor necrosis factor (TNF)-α, leukotriene B4 (LTB4) and 6-keto-PGF-1α (degradation product of PGI2) in the gastric tissue. RESULTS: Our data showed that C. butyricum significantly reduced the gastric mucosal injury area and ameliorated the pathological conditions of the gastric mucosa. C. butyricum not only minimized the decreases in activity of SOD and CAT, but also reduced the level of MDA in all three GU models used in this study. The accumulation of IL1-ß, TNF-α and LBT4 decreased, while 6-keto-PGF-1α increased with pretreatment by C. butyricum in all three GU models. CONCLUSION: Our data demonstrated the protective effects of pretreatment with C. butyricum on anti-oxidation and anti-inflammation in different types of GU models in mice. Further studies are needed to explore its potential clinical benefits.

[Effects of ambroxol combined with low-dose heparin on TNF-alpha and IL-1beta in rabbits with acute lung injury].

OBJECTIVE: To investigate the intervention and mechanism of ambroxol combined with low-dose heparin on oxidative stress, TNF-alpha and IL-1beta in rabbits with acute lung injury (ALI). METHODS: Twenty-four healthy Japanese rabbits were randomly divided into three groups: (1) Normal saline control group (NC), (2) Oleic acid injury group (OA), (3) Ambroxol + low-dose heparin therapy group (AH). After the success of ALI model, AH group was injected ambroxol + low-dose heparin, while the NC group and OA group were injected the same dose of normal saline by the same method. Arterial oxygen tension (PaO2), tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) at different time points were determined. The pathological manifestation of both side lungs was observed at the end of expeiment. The activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), xanthine oxidase (XO) and the content of malondialdehyde (MDA) in bronchoalveolar lavage fluid (BALF) and lung tissue homogenate were tested. The apoptosis index was detected. The lung wet/dry (W/D) ratio was calculated. The pathological changes in lung tissue were observed by light microscopy, and the ultrastructural changes of lung tissue were observed by electron microscopy. RESULTS: (1) The instructive injury induced by ALI observed under electron microscope and light microscope and W/D was decreased significantly in AH group. (2) PaO2 was improved significantly in AH group, compared with that in OA group (P < 0.01). (3) The activity of GSH-Px and SOD in AH group increased significantly (P < 0.01 or P < 0.05) but the activity of XO and the content of MDA decreased significantly (P < 0.01), compared with those in OA group. (4) Except the content of IL-1beta in serum before treatment, the content of IL-1beta and TNF-alpha in serum, BALF, lung tissue homogenate of OA group increased significantly (P < 0.01), and those were obviously improved in AH group. (5) Apoptosis index (AI) in AH group decreased significantly (P < 0.01) compared with that in OA group. CONCLUSION: In ALI induced by OA, IL-1beta and TNF-alpha increases significantly and involved in the occurrence and development of ALI. Ambroxol combined with low-dose heparin can reduce lung cells oxidative stress to inhibit the release of IL-1beta and TNF-alpha, which play a role in the treatment of ALI.

Acupressure therapy inhibits the development of diabetic complications in Chinese patients with type 2 diabetes.

OBJECTIVES: The objectives of this study were to investigate the effects of acupressure therapy (AT) on the development and progression of diabetic complications in Chinese patients with type 2 diabetes (T2D). DESIGN AND METHODS: A total of 80 patients with T2D were recruited for a randomized clinical study of the effect of AT on the progression and development of diabetic complications, and 64 patients were followed up for 3 years. All patients with T2D were treated with regular medicines and participated in diet and exercise programs for the control of hyperglycemia and hypertension. The patients in the AT group received additional treatment of a 90-minute AT 4-6 times per week for 3 consecutive years. Their blood lipids, fasting glucose levels, and heart and kidney functions and nerve conduction velocity (NCV) were longitudinally monitored before and every 12 months after AT. RESULTS: Following AT therapy for 3 years, significantly lower levels of total cholesterol, triglycerides, low-density lipoprotein-cholesterol, and higher levels of high-density lipoprotein-cholesterol (HDL-C) were observed and no significantly increased levels of serum creatinine and urine protein were detected in the AT group, as compared with that in controls. Furthermore, the mean values of NCV in the AT group at 2 years post-treatment were significantly greater than those of controls and were further elevated at the end of this study. Therefore, AT inhibited the progression of hyperlipidemia and improved diabetes-associated kidney function and neuropathy in Chinese patients with T2D. CONCLUSIONS: AT may be an effective nonpharmacological adjunctive strategy for alleviating the development and progression of T2D-related complications.

[Effect of Safflor injection on the intestine ultrastructure characteristics following intestine ischemia/reperfusion injury in rabbits].

AIM: To explore effects of Safflor (Chinese Tradional Medicine) on the intestine ultrastructure characteristics during intestine ischemia/ reperfusion injury (I/RI) in rabbits. METHODS: Thirty rabbits were randomly divided into three groups: control group (group S), ischemia/reperfusion group (group I/R) and Safflor injection group (group SI). Morphological changes of intestine ischemia/reperfusion in rabbits and the protective effects of Safflor were observed under electric telescope. RESULTS: The intestine ultrastructure was badly injured in group I/R. Mitochondria and intestinal mucosal cells were swellen and endoplasmic reticulum expanded, however, in the SI group the ultrastructural injury of the ischemia greatly ameliorated. CONCLUSION: The ultrastructure injury occurrted after intestine I/RI and Safflor has protective effects on the intestine ultrastructure.

[Protective effects of polydatin against lung ischemia/reperfusion injury and the initial exploration for its mechanism].

AIM: To observe protective effects of polydatin (PD) during lung ischemia/reperfusion injury (LI/RI) and investigate its potential mechanism . METHODS: Rabbit lung model of ischemia/reperfusion injury was constituted in vivo. The 40 rabbits were randomly divided into four groups (n = 10): control group (C group), ischemia/reperfusion group (I/R), PD + I/R group (PD) and PD+ polymyxin B (PMB) + I/R group (PMB). The blood specimen gathered at different time points were tested for the content of melondialdehyde (MDA) and the enzyme activity of superoxide dismutase (SOD). The lung tissue sampled at the end of the experiment were assayed for wet/dry weight ratio (W/D), injured alveoli rate (IAR) and observing ultrastructure changes under electron micro scope. RESULTS: (1) The activity of SOD showed a similar time-dependent decline in I/R group and PMB group during I/R, while in PD group this tendency was milder (P < 0.01 vs I/R group). (2) In contrast to the results above, the level of MDA markedly increased in I/R and PMB group, but was slowed down in PD group (P < 0.01 vs I/R group). (3) The value of W/D) and IAR was much higher in I/R and PMB group (P < 0.05 or P < 0.01 vs C group). In PD group, it was decreased (P < 0.01 vs I/R group or PMB group). (4) Electron microscope showed obvious ultrastructure injury brought by LI/RI in I/R group and PMB group, which was greatly attenuated in PD group. CONCLUSION: PD can protect lung from LI/RI, and PKC may participate in its mechanisms.

[Effects of safflower injection on cycloxygenase in rabbits lung ischemia/reperfusion injury].

AIM: To observe protective effects of safflower injection (SI) on lung ischemia/reperfusion injury (LIRI) and investigate its potential mechanism. METHODS: Rabbit lung model of ischemia/reperfusion injury was constituted in vivo. The rabbits were randomly divided into three groups: sham-operation group (S group), ischemia/reperfusion group (I/R group) and ischemia/reperfusion plus safflower injection group (SI group). Malondialdehyde (MDA) content, superoxide dismutase (SOD) and xanthine oxidase (XO) activities in serum were measured. The lung tissue sampled at the end of the experiment was assayed for wet/dry weight ratio (W/D), injured alveoli rate (IAR) and ultrastructural changes were observed under electron microscope. The expression of COX-1 and COX-2 were measured by immunohistochemistry (IHC). The expressions of COX-1mRNA and COX-2mRNA were observed by in situ hybridization (ISH). RESULTS: In I/R group, XO and MDA increased and SOD decreased in serum, while the same changes happened in SI group but less severely(P<0.01). The value of W/D and IAR was much higher in I/R group than S group, but decreased in SI group. Electron microscope showed obvious ultrastructural injury brought by LIRI in I/R group, which was greatly attenuated in SI group. The IHC and ISH demonstrated that COX-2 and COX-2mRNA in pulmonary tissue of I/R group were significantly higher than those of SI group (P < 0.01). The difference of COX-1 and COX-1mRNA in pulmonary tissue among the three groups was not significant. CONCLUSION: The ischemia/reperfusion lung injury insults induced the regulation of COX-2 in lung. Safflower injection may attenuate lung ischemia/reperfusion injury through inhibiting cyclooxygenase-2 expression.

[Effect of ligustrazine on expression of Fas/FasL in pulmonary injury induced by ischemia/reperfusion in rabbits].

AIM: To investigate the effect of ligustrazine (LGT) on expression of Fas/FasL mRNA during pulmonary ischemia/reperfusion injury (PI/RI) in the rabbits. METHODS: Single lung ischemia/reperfusion animal model was used in this study. The rabbits were randomly divided into three groups (n = 30, in each): sham operated group (Sham), I/R group (I/R) and I/R + LGT group (I/R + LGT). Changes of several parameters which included apoptotic index (AI), wet to dry ratio of lung tissue weight (W/D) and index of quantitative assessment of histologic lung injury (IQA) were measured at 1h, 3h, 5h after reperfusion in lung tissue. Meanwhile the location and expression of Fas/FasL mRNA were observed. Lung tissue was prepared for light microscopic and electron microscopic ob servation at 1 h, 3 h, 5 h after reperfusion. RESULTS: As compared with group I/R, Fas/FasL mRNA slightly expressed in intima and extima of small pulmonary artery, alveoli, and bronchiole epithelia in group LGT. The values of AI, W/D and IQA showed significantly lower in group I/R + LGT than that in group I/R at 1 h, 3 h, 5 h after reperfusion in lung tissue (P < 0.01 and P < 0.05). Meanwhile, abnormal changes of the lung tissue in morphologically were lessen markedly in group I/R + LGT. CONCLUSION: Ligustrazine has notable protective effects on PI/RI in rabbits by inhibiting Fas/FasL mRNA express in lung tissue and decreasing apoptosis.

[Effect of ligustrazine and L-arginine on function of mitochondria in myocardium after ischemia/reperfusion injury of myocardium in rabbits].

AIM: To study the effect of ligustrazine (LGT) and L-arginine(L-Arg)on function of mitochondria in myocardium after myocardial ischemia/reperfusion injury (MI/RI). METHODS: 50 rabbits were randomly divided into five groups (n=10): Control group(A), MI/R group(B), MI/R + LGT group (C), MI/R+ L-Arg group (D), MI/R+ LGT + L-Arg group (E). The mitochondrial respiratory function, Ca2+ concentration ([Ca2+]m), malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were deter mined. Meanwhile, the contents of ATP and EC in the myocardial tissue were measured, respectively. RESULTS: It was found that mitochondrial respiratory control rate (RCR), state 3 (ST3), SOD in C, D, E group were higher than those of B group, state 4 (ST4), [Ca2+]m, MDA were lower than those of B group, ATP and EC levels of myocardial tissue were higher than those in B group; and there were not significant differences between E and A group of above. CONCLUSION: LGT and IL-Arg can improve function of mitochondria in myocardium after ischemia/reperfusion injury of myocardium in rabbits by decreasing oxygen free radical level and Ca" overload in the mitochondria.