Fibroblast growth factor -21 (FGF-21) is a recently discovered metabolic regulation factor, regulating glucose and lipid metabolism and increasing insulin sensitivity. FGF-21 is expected to be a potential anti-diabetic drug. Expression of FGF-21 as inclusion bodies has advantages for high yield and purity, but the bioactivity of the protein is almost totally lost after denature and renature. That is why FGF-21 is currently expressed in soluble form. As a result, the yield is considerably low. In this study, we used SUMO vector to express SUMO-human FGF-21 (SUMO-hFGF-21) in form of inclusion body. We optimized the culture conditions to increase the yield of the bioactive human fibroblast growth factor-21. We applied the hollow fiber membrane filtration column to enrich the bacteria, wash, denature and renature inclusion bodies. After affinity and gel filtration chromatography, we examined the hypoglycemic activity of FGF-21 by the glucose uptake assay in HepG2 cells. We also detected the blood glucose concentration of type 2 diabetic db/db model mice after short or long-term treatment. The results show that the yield of ihFGF-21 was 4 times higher than that of shFGF-21. The yield was 20 mg/L for ihFGF-21 vs. 6 mg/L for shFGF-21. The purity of ihFGF-21 was above 95%, while shFGF-21 was 90%. Compared with the traditional method of extracting inclusion bodies, the production cycle was about three times shortened by application of hollow fiber membrane filtration column technology, but the bioactivity did not significantly differ. This method provides an efficient and cost-effective strategy to the pilot and industrial production of hFGF-21.
Animals ; Bacteria ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Disease Models, Animal ; Fibroblast Growth Factors ; biosynthesis ; Genetic Vectors ; Glucose ; metabolism ; Hep G2 Cells ; Humans ; Hypoglycemic Agents ; isolation & purification ; Inclusion Bodies ; metabolism ; Mice ; Recombinant Fusion Proteins ; biosynthesis ; Small Ubiquitin-Related Modifier Proteins ; biosynthesis

AIM: To investigate the effect of ambroxol on pulmonary and vascular injury in chronically hypoxic rats. METHODS: 36 male Wistar rats were randomly divided into 3 groups: normal control,chronically intermittent hypoxia(CIH) and ambroxol precaution group(AP).The CIH and AP groups were made into the chronically hypoxic models .The mean pulmonary artery pressure(PAPM) and the levels of plasma superoxide dismutase (SOD) and plasma nitric oxide (NO),lipid peroxide(LPO) were determined. The levels of the lung homogenates SOD, LPO, NO and the changes in pulmonary vascular structure were also examined. RESULTS: The levels of plasma and lung homogenates SOD,NO in CIH group were respectively significantly lower than that of normal control and AP group ( P

AIM: To establish a guinea pig asthma model and to evaluate the effect of airway remodeling on airway responsiveness. METHODS: The guinea pig asthma model was established by ovalbumin (OVA) sensitization and challenge repeatedly. Bronchial provocation tests were conducted through intravenous injection of acetylcholine. The airway morphologic parameters were measured by computer image analysis system. White blood cells and the differential count in bronchoalveolar lavage fluid (BALF) were examined. RESULTS: The resistance of airway was increased significantly after 4 weeks of OVA exposure, but the increase disappeared upon prolonged exposure. After 8 weeks of OVA exposure, fiber tissue in large airway was increased, and the thickness of smooth muscle layer of small airway was enlarged, as compared with that in control animals. CONCLUSION: Airway responsiveness has changed after prolonged OVA exposure in guinea pigs. This change is related to airway remodeling. [

AIM: To reproduce hypercapnic models and approach some pathophysiological changes in rats. METHODS: The mixed gases of high concentrated carbon dioxide (8% CO 2, 21% O 2, 71% N 2) were given to wistar rats 7 hours a day for 28 days. The various indexes were compared between control group (group A) and hypercapnic group (group B). RESULTS: The PaCO 2 [(55.90?4.34) mmHg] and the lipid peroxides (LPO) contents in plasma, lung tissue and right ventricle were significantly higher in group B than those in group A ( P

AIM: To investigate the effects of chronic hy poxia and antagonistic effects of aminophylline on airway inflammation and oxida tive lung damage in rats. METHODS: Thirty-four male Wistar rats were randomly divided into three groups: normal control group (n=10); hypoxia group (n=12); aminop hlline-treated group (n=12). The last two groups were both exposed to hypoxi a 7 hours per day for 21 days. The third group was treated with aminophlline (1 00 mg?kg -1?d -1) before exposed to hypoxia. The level of tumor ne c rosis factor (TNF) -?, interleukin (IL)-10, lipid peroxide (LPO) and the activi ty of superoxide dismutase (SOD) were determined in blood and homogenates of lung tissue. RESULTS: Compared to the control group, the levels of TNF-?, IL -10 and LPO were significantly increased (P

OBJECTIVETo develop the preparation method of licorice juice as a processing excipient and to determine the proper proportion of licorice juice and fructus evodiae during process.

METHODNine experiments were carried out through L9 (3(4)) orthogonal design. The wetting results were compared by using different proportion of licorice juice and fructus evodiae.

RESULTThe optimal preparation process was as follows: the licorice roots were cutted into slices and 12 times of water was added. Then they were decocted for 1 h. The second and third decoction were also carried out by 12 times of water and 1 h. The proper proportion of licorice juice and fructus evodiae was 1:1.

CONCLUSIONThe preparation process is scientific and suitable to prepare the processing excipient.

Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Evodia ; chemistry ; Excipients ; chemistry ; Glycyrrhiza ; chemistry

Previous studies proposed that the synergistic effect of fibroblast growth factor-21 (FGF-21) and insulin may be due to the improvement of insulin sensitivity by FGF-21. However, there is no experimental evidence to support this. This study was designed to elucidate the mechanism of synergistic effect of FGF-21 and insulin in the regulation of glucose metabolism. The synergistic effect of FGF-21 and insulin on regulating glucose metabolism was demonstrated by investigating the glucose absorption rate by insulin resistance HepG2 cell model and the blood glucose chances in type 2 diabetic db/db mice after treatments with different concentrations of FGF-21 or/and insulin; The synergistic metabolism was revealed through detecting GLUT1 and GLUT4 transcription levels in the liver by real-time PCR method. The experimental results showed that FGF-21 and insulin have a synergistic effect on the regulation of glucose metabolism. The results of real-time PCR showed that the effective dose of FGF-21 could up-regulate the transcription level of GLUT1 in a dose-dependent manner, but had no effect on the transcription level of GLUT4. Insulin (4 u) alone could up-regulate the transcription level of GLUT4, yet had no effect on that of GLUT1. Ineffective dose 0.1 mg kg(-1) FGF-21 alone could not change the transcription level of GLUT1 or GLUT4. However, when the ineffective dose 0.1 mg x kg(-1) FGF-21 was used in combination with insulin (4 u) significantly increased the transcription levels of both GLUT1 and GLUT4, the transcription level of GLUT1 was similar to that treated with 5 time concentration of FGF-21 alone; the transcription level of GLUT4 is higher than that treated with insulin (4 u) alone. In summary, in the presence of FGF-21, insulin increases the sensitivity of FGF-21 through enhancing GLUT1 transcription. Vice versa, FGF-21 increases the sensitivity of insulin by stimulating GLUT4 transcription in the presence of insulin. FGF-21 and insulin exert a synergistic effect on glucose metabolism through mutual sensitization.

OBJECTIVETo investigate the protective effects of beta-carotene in rats against the development of chronic bronchitis induced by cigarette smoking.

METHODSForty-two Male Wistar rats were randomly divided into three study groups: (1) control (n = 15), animals underwent no treatment; (2) cigarette smoking (n = 15), animals developed chronic bronchitis through long-term cigarette smoking twice a day for 75 d; (3) beta-carotene plus cigarette smoking animals (n = 12) were given 1 ml or 15 mg/kg beta-carotene orally every day just before cigarette smoking. The levels of IL-6, IL-8, NO, superoxide dismutase (SOD) and lipoperoxide (LPO) in serum, bronchoalveolar lavage fluid (BALF) and lung tissue were measured and the pathological changes to lung tissue were analyzed using light microscopy.

RESULTSLong-term cigarette smoking caused an obvious increase in the amount of IL-6, IL-8 and LPO and a sharp decrease in the levels of NO and SOD in smoking animals compared to controls. beta-carotene intake reversed all the changes induced by smoking and alleviated the pathological changes caused by chronic bronchitis.

CONCLUSIONSQuantitative oral intake of beta-carotene had protective effects against chronic bronchitis induced by long-term cigarette smoking, which was associated with the increased production of NO, the clearance of some oxidative free radicals (OFR) and the alleviation of chronic inflammation.

Animals ; Bronchitis ; blood ; etiology ; prevention & control ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Male ; Nitric Oxide ; blood ; Rats ; Rats, Wistar ; Smoking ; adverse effects ; Superoxide Dismutase ; blood ; beta Carotene ; pharmacology

AIM: To explore the effect of hydrogen sulfide on the senescence of human umbilical vein endothe -lial cells (HUVECs) induced by high glucose.METHODS: Senescence model was established by treating HUVECs with33 mmol/L glucose for 48 h.The parameters were detected to demonstrate the effect of hydrogen sulfide on senescence andthe mechanism involved was also investigated .RESULTS: In the cells treated with high glucose, the proliferation was attenuatedwith a higher number of senescence -associated β-galactosidase (SA-β-Gal) positive cells, and plasminogen activatorinhibitor 1 (PAI-1) protein expression, malondialdehyde (MDA) production and NF-κB p65 activity were increasedsignificantly, but the expression of superoxide dismutase 1 (SOD1) was decreased.However, the cell number and SOD1expression were increased, and the number of SA-β-Gal positive cells, PAI-1 protein expression, MDA production and theactivity of NF-κB p65 were decreased after sodium hydrosulfide (100 and 200 μmol/L) treatment.CONCLUSION: Exogenoushydrogen sulfide prevents HUVECs against high glucose -induced senescence by suppressing oxidative stress and NF -κB p65 activity.