Objective The purpose of this study was to prepare RGD4C modified ferritin nanocages (RGD4C-FRT) for targeted drug delivery to rat hepatic stellate cells (HSC-T6). Methods RGD4C modified human H-chain ferritin was expressed and purified. Doxorubicin (Dox) was encapsulated into the cavity of RGD4C-FRT through ion channels, which resulted in RGD4C-FRT-Dox. The target of RGD4C-FRT-Dox to HSC-T6 was detected using fluorescence microscopy. Results Transmission electron microscopy showed that RGD4C-FRT was hollow spherical-structured with uniform size and good dispersion. The average particle diameters of RGD4C-FRT and RGD4C-FRT-Dox were 12.57 nm and 20.13 nm , respectively. The drug encapsulation efficiency and loading percentage of RGD4C-FRT-Dox were 77.32% and 15.88% respectively. RGD4C-FRT-Dox was significantly uptaken by HSC-T6, and the uptake could be blocked by the empty carrier RGD4C-FRT. Conclusion RGD4C-FRT-Dox can specifically target HSC-T6. Further animal experiments are needed to inspect its therapeutic effect on liver fibrosis.

OBJECTIVETo investigate the effect of siRNA silencing the role of C-Jun N-terminal Kinase (JNK) gene in excessive endoplasmic reticulum stress on lung ischemia/reperfusion injury.

METHODSMouse model of pulmonary ischemia reperfusion injury (PIRI) in situ was established with unilateral lung in vivo. Seventy experimental mice were randomly allocated into seven groups (n = 10): Sham group (Sham group), ischemia reperfusion group (I/R), PBS+ Lipofectamine2000TM transfection reagent group (I/R + PBS+ Lipo group), negative control group (I/R+ SCR group), JNK-siRNA group (I/R + siRNA(JNK1), siRNA(JNK2), siRNA(JNK3)). Mice were euthanized after experimental time out, and left lung tissue was extracted. Wet/dry lung weight ratio (W/D) and total lung water content (TLW) were tested. Light microscope, alveolar damage quantitative evaluation index (IQA) and electron microscope were observed. The expression levels of JNK and glucose regulatex protein(GRP78) were detected by RT-PCR and Western blot. Apoptosis of lung tissue was determined by TUNEL.

RESULTSCompared with Sham group, all indicators above of I/R + PBS + Lipo group and I/R + SCR group were significantly increased (P < 0.01), and compared with I/R group, those indicators of the three groups all had no notable difference; those indicators were not statistically different between I/R + PBS + Lipo group and I/R + SCR group, and compared to the three groups, the above indicators in JNK-siRNA group were lower (P < 0.05, P < 0.01) except that the expression levels of GRP78 was not statistically different.

CONCLUSIONI/R induces excessive ERS in lung tissue, in which JNK pathway participates in apoptosis, leading to lung tissue injury.

Animals ; Apoptosis ; Endoplasmic Reticulum Stress ; Heat-Shock Proteins ; metabolism ; JNK Mitogen-Activated Protein Kinases ; genetics ; Lung ; physiopathology ; Lung Injury ; genetics ; MAP Kinase Signaling System ; Mice ; RNA, Small Interfering ; Reperfusion Injury ; genetics

AIMTo study the effect of diabetes-like environment on the cardiac hypertrophy, cultured cardiomyocytes were used to study the effect of high insulin and high glucose on norepinephrine (NE)-induced cardiac hypertrophy.

METHODSUsing cultured myocardial cells as a model, the cellular hypertrophy was observed. The contracting frequency was counted by the inverted microscope, the protein content was assayed with Lowry's method, the cardiomyocytes' volumes were measured by computer photograph analysis system, the protein synthesis was assayed with [3H] leucine intake method.

RESULTSThe total cellular protein content, cellular volumes, cellular protein synthesis showed an increase in high insulin group and high glucose group compared with control group. High insulin and high glucose and NE group showed a further increase compared with high glucose and NE group.

CONCLUSIONThe high insulin itself induces hypertrophy of the cultured myocardial cells slightly. Meanwhile, imitating diabetes-like environment with high insulin and high glucose and NE can further accelerate hypertrophy of the cultured myocardial cells.

Animals ; Animals, Newborn ; Cardiomegaly ; chemically induced ; metabolism ; Cells, Cultured ; Glucose ; metabolism ; Insulin ; pharmacology ; Myocytes, Cardiac ; drug effects ; metabolism ; Norepinephrine ; adverse effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo detect the self-assembly morphology of sodium hyaluronate injection on mica using atomic force microscopy(AFM).

METHODSAtomic force microscopy with nanometer resolution was used to observe the self-assembly morphology of different concentrations of sodium hyaluronate injection on mica at room temperature.

RESULTSThe self-assembly morphology of 0.001, 0.01, and 0.1 mg/ml sodium hyaluronate injection on mica featured piebald, reticular and dendritic structures, respectively. At 1 and 5 mg/ml, sodium hyaluronate injection displayed bacilliform and spherical structures on mica, respectively; the diameter and height of the particles of 5 mg/ml sodium hyaluronate was 197.97±78.48 nm and 30.79±18.67 nm, significantly greater than those of 0.1 mg/ml sodium hyaluronate injection (49.52±11.93 nm and 5.37±1.59 nm, respectively, P<0.05).

CONCLUSIONThe self-assembly morphology of sodium hyaluronate injection on mica varies with its concentration. The piebald and reticular structure may facilitate the function of sodium hyaluronate, and the dendritic feature resembles the representative model of diffusion-limited aggregation (DLA).

Aluminum Silicates ; chemistry ; Hyaluronic Acid ; administration & dosage ; chemical synthesis ; chemistry ; Microscopy, Atomic Force ; Nanostructures ; Surface Properties

Objective:To study the effect of curcumin on enterocyte apoptosis and its protective effect on intestinal mucosal barrier in septic rats.Methods:Eighty-seven 3-month male Sprague-Dawley (SD) rats were divided into Sham group, model group and curcumin group by random number table method, with 29 rats in each group. The septic rat model was reproduced by cecal ligation and puncture (CLP). 4 mL dimethyl sulfoxide solution were intraperitoneally injected in both Sham group and model group, 200 mg/kg curcumin dissolved by 4 mL dimethyl sulfoxide solution were intraperitoneally injected in curcumin group 10 minutes after operation. The blood samples (15 rats in each group) were collected 2, 12, 24 hours after operation, and the levels of serum procalcitonin (PCT), tumor necrosis factor-α(TNF-α), D-lactic acid and diamine oxidas (DAO) were tested by enzyme linked immunosorbent assay (ELISA). The ileum tissues were collected 12 hours, 24 hours after operation in three groups, water content was tested by weighting, pathologic structure was observed by light microscope, the enterocyte apoptosis was tested by terminal deoxynucleotidyl transferase-mediated dUTP-nick end labeling method (TUNEL). The 7-day survival rate was observed in three groups (14 rats in each group).Results:The serum levels of PCT, TNF-α, D-lactic acid and DAO were higher in model group at 2, 12, 24 hours after operation than those in Sham group, PCT, TNF-α levels were significantly higher in model group than those in Sham group 2 hours after operation [PCT (μg/L): 1.89±0.17 vs. 0.10±0.02, TNF-α (ng/L): 216.51±1.47 vs. 85.25±8.20, both P < 0.01], D-lactic acid, DAO levels were significantly higher in model group than those in Sham group 12 hours after operation [D-lactic acid (mg/L): 40.53±7.76 vs. 11.29±1.28, DAO (ng/L): 1 120.40±302.35 vs. 330.02±81.28, both P < 0.01]. Compared with model group, the levels of serum PCT, TNF-α, D-lactic acid and DAO were lower in curcumin group 2, 12, 24 hours after operation, the statistical difference appeared from 12 hours after operation [PCT (μg/L): 5.37±0.44 vs. 8.67±0.64, TNF-α(ng/L): 211.12±4.31 vs. 313.30±18.46, D-lactic acid (mg/L): 29.74±1.41 vs. 40.53±7.76, DAO (ng/L): 810.71±201.41 vs. 1 120.40±302.35, all P < 0.05], curcumin group had lower water content in ileum tissues 12 hours, 24 hours after operation [(68.34±0.68)% vs. (70.55±0.87)%, (69.41±0.59)% vs. (71.69±0.87)%, both P < 0.05]. The pathologic structures of intestinal villus were normal in Sham group, however, in model group intestinal villus were atrophic, edematous and shorten 12 hours after operation, it was further exacerbated 24 hours after operation. Compared with model group, the pathologic structures of intestinal villus in curcumin group were relived 12 hours, 24 hours after operation. The number of apoptotic enterocytes were significantly increased in model group compared with Sham group 24 hours after operation (cells: 25.48±6.10 vs. 4.00±2.04, P < 0.05), and the number of apoptotic enterocytes was lower in curcumin group than that in model group at the same time (cells: 15.48±3.75 vs. 25.48±6.10), the difference was statistically significant (both P < 0.05). Seven-day survival rate was significantly lower in curcumin than that in model group [42.9% (6/14) vs. 50.0% (7/14)], however, the difference was not statistically significant ( P > 0.05). Conclusion:Curcumin can protect the intestinal mucosal barrier by inhibiting enterocyte apoptosis in septic rats.

Objective To calculate the delay times and embedding dimensions of surface electromyogragh(sEMG) signal of the masseter and temporal muscle during clenching at intercuspal position (ICP) , and to reconstruct the space phase of sEMG signal with method of nonlinear dynamics.Methods Ten male and 10 female young volunteers with normal masticatory system were included in the study, the signals of surface sEMG of bilateral masseter and temporal muscles in the position of ICP were collected, and the delay times and embedding dimensions of the sEMG signals were calculated using C-C method, the space phase of the sEMG signals was reconstructed.Results A program of nonlinear dynamic analysis of mastication muscles sEMG signals was developed base on the platform of Matlab.The signals of the sEMG of the masseters and temporals were analyzed with nonlinear dynamic methods, the delay times and embedding dimensions of each muscles sEMG signals were obtained, and the space phase of each sEMG signals were reconstructed.The graphs of the space phase indicated the typical characteristic of chaotic attractor.Conclusions The signals of the masseter and temporal muscles of surface sEMG during clenching at ICP of normal masticatory system show the typical characters of chaos, the methods of nonlinear dynamics are effective to analyze the sEMG signals of mastication system.

Fibroblast growth factor 21 (FGF21) is a member of FGF family. It has been demonstrated that FGF21 is an independent, safe and effective regulator of blood glucose levels in vivo. In order to improve the activity of FGF21, we exchanged the beta10-beta12 domain of the human FGF21 with that of the mouse FGF21 to construct a novel FGF21 gene (named hmFGF21), and then subcloned hmFGF21 gene into the SUMO expression vector to create pSUMO-hmFGF21 and transformed it into E. coli Rosetta for expression of the fusion protein SUMO-hmFGF21. Both in vitro and in vivo glucose regulation activity of hmFGF21 was evaluated. The SDS-PAGE result showed that compared with wild-type hFGF21, the soluble expression of hmFGF21 increased about 2-fold. HmFGF21 was more potent in stimulation of glucose uptake in HepG2 cells in vitro. The results of anti-diabetic effect on db/db mice demonstrated that hmFGF21 had better efficacy on controlling the blood glucose of the db/db diabetic animals than wild-type hFGF21. These results suggest that the biological properties of FGF21 are significantly improved by optimization.
Amino Acid Sequence ; Animals ; Blood Glucose ; metabolism ; Cysteine Endopeptidases ; Diabetes Mellitus, Experimental ; blood ; Endopeptidases ; genetics ; Escherichia coli ; Fibroblast Growth Factors ; genetics ; metabolism ; pharmacology ; Genetic Vectors ; Glucose ; metabolism ; Hep G2 Cells ; metabolism ; Humans ; Hypoglycemic Agents ; metabolism ; pharmacology ; Male ; Mice ; Mutation ; Plasmids ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Transformation, Genetic

OBJECTIVETo evaluate the effect of water pollution with dexamethasone on intestinal flora in mice.

METHODSTwenty Balb/c mice were randomly divided into control group and low-, moderate- and high-dose dexamethasone groups. The mice in dexamethasone groups were exposed to dexamethasone sodium phosphate in drinking water at doses of 0.035, 0.225, and 2.25 ng for 36 days. The changes in behaviors, fur condition, and feces of the mice were observed daily. All the mice were sacrificed at 36 days and the tissues in the ileocecal region was collected for denaturant gradient gel electrophoresis (DGGE) of 16S rDNA V6 variable regions of microbes and sequence analysis with BLAST.

RESULTSThe mice in the 3 dexamethasone groups all showed aggressive behaviors. Cluster analysis of DGGE graph showed relatively stable floras in the ileocecal region in all the mice, but principal component analysis identified differences in the dominating flora among the groups. Diversity analysis of the flora revealed significantly increased amount and types of bacteria in the intestinal flora in all the 3 dexamethasone groups (P<0.05 or 0.01) compared with the control group. Sequence analysis of 16S rDNA V6 regions showed 15 common bacterial species and 2 differential species between the dexamethasone groups and the control group with changes in the type and proportion of the dominating bacterium in the dexamethasone groups. Lactobacillus colonization was detected in the control group but not in moderate- and high-dose dexamethasone groups, and Shigella species were found in the latter two groups.

CONCLUSIONSWater contamination with dexamethasone can affect the nervous system of mice, cause changes in the types and amounts of intestinal bacteria and the dominating bacteria, and inhibit the colonization of probiotics in the intestinal floras to increase the risk of invasion by intestinal pathogenic bacteria.

Animals ; Bacteria ; classification ; Dexamethasone ; pharmacology ; Drinking Water ; chemistry ; Feces ; Gastrointestinal Microbiome ; drug effects ; Lactobacillus ; isolation & purification ; Mice ; Mice, Inbred BALB C ; Probiotics ; RNA, Bacterial ; genetics ; RNA, Ribosomal, 16S ; genetics ; Shigella ; isolation & purification

OBJECTIVETo compare the bidirectional effect of rhubarb total anthraquinone (TA) and total tannins (TT) on rats' liver.

METHODSOne hundred rats were randomly divided into 10 groups, i.e., the blank group, the model group, the blank + high dose TA group, the blank +low dose TA group, the blank + high dose TT group, the blank + low dose TT group, the model + high dose TA group, the model + low dose TA group, the model +high dose TT group, and the model + low dose TT group, 10 in each group. The carbon tetrachloride (CCI4) was used to prepare the acute liver injury rat model. TA and TT of rhubarb (at 5.40 g crude drugs/kg and 14.69 g crude drugs/kg) were intragastrically administrated to rats in all groups except the blank group and the model group, once daily for 6 successive days.The general state of rats, biochemical indices such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), laminin (LN), hyaluronic acid (HA), transforming growth factor beta1 (TGF-beta1), as well pathological results of rat liver tissues. Finally the protection laws of TA and TT for rats' liver were analyzed using factor analysis.

RESULTSCompared with the blank control group, all biochemical indices increased in the blank group (P < 0.05, P < 0.01). HA also increased in the blank + high dose TA group; AST, ALT, and HA also increased in the blank +high dose TT group (P < 0.05). Compared with the model group, AST, ALT, ALP, HA, and TGF-beta1 significantly decreased in the model + low dose TA group, the model + high dose TA group, the model + low dose TT group (P < 0.05, P < 0.01). Serum AST, ALT, and ALP also decreased in the model + high dose TT group (P < 0.05, P < 0.01). Pathological results showed that mild swollen liver cells in the model + high dose TA group. Fatty degeneration and fragmental necrosis around the central veins occurred in the blank + high dose TA group. The pathological injury was inproved in the model +low dose TA group. Two common factors, liver fibrosis and liver cell injury, were extracted by using factor analysis. TA showed stronger improvement of the two common factors than TT.

CONCLUSIONSRhubarb TA and TT showed protective and harmful effects on rats' liver. At an equivalent dosage, TA had better liver protection than TT. High dose TT played a role in liver injury to some extent.

Animals ; Anthraquinones ; adverse effects ; pharmacology ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; drug therapy ; pathology ; Dose-Response Relationship, Drug ; Female ; Liver ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry ; Tannins ; adverse effects ; pharmacology

Objective: To observe the clinical efficacy of ginger-partitioned moxibustion plus pediatric massage (tuina) in treating infantile diarrhea due to spleen deficiency. Methods: Ninety infants were randomly divided into a massage plus moxibustion group, a massage group and a drug group by the random number table method, with 30 cases in each group. The intervention was conducted for two consecutive courses. The infants in the massage plus moxibustion group were treated with pediatric massage and ginger-partitioned moxibustion at Shenque (CV 8). The infants in the massage group were treated with pediatric massage alone, while those in the drug group were treated with smecta. The primary and secondary symptom scales were assessed before and after treatment and at the follow-ups, and the total effective rate was evaluated after treatment. Results: The total effective rate in the massage plus moxibustion group was significantly different from that in the massage group and drug group (both P<0.05). After treatment, the scores of primary and secondary symptoms decreased in all three groups, with statistically significant intra-group differences (all P<0.05); the scores of primary symptoms were significantly different between the massage plus moxibustion group and the drug group (P<0.05); the scores of secondary symptoms in the massage plus moxibustion group and the massage group were significantly different from that in the drug group (both P<0.05). The differences in the time to recover normal bowel movement frequency among the three groups were not statistically significant (P>0.05). Conclusion: Ginger-partitioned moxibustion plus pediatric massage compared with pediatric massage or smecta monotherapy shows superior clinical efficacy in treating infantile diarrhea due to spleen deficiency, and has the advantages of appetite improvement, physique strengthening and short course.