Objective:To study the effects of diosgenin on cell proliferation, differentiation and OPG/RANKL mRNA expression of rat osteoblasts cultured in vitro for analyzing the prevention and treatment mechanism of diosgenin. Methods: Rat osteoblast cultured in vitro was treated by a series of different concentrations of diosgenin for different time respectively. The proliferation of osteoblast was detected by MTT method. The differentiation of osteoblast was detected by the activity of ALP mRNA expression of osteoprotegerin(OPG) and RANKL(receptor activator nuclear factor-kappa B ligand,RANK) were semi-quantified by RT-PCR, and the ratio of OPG/RANKL mRNA was used to evaluated the effect of diosgenin on osteoclast signal transduction pathway of OPG/RANKL/RANK system. Results: Diosgen at the concentration of 3.64?10-8 and 3.64?10-7mol/L can promote the proliferation, enhance the activity of ALP and raise the ratio of OPG/RANKL mRNA of rat osteoblast cultured in vitro(P

The effects of magnolol (Mag) on hyperglycemia and hyperlipemia, hepatic oxidative stress and cytochrome P4502E1 (CYP2E1) activity of diabetic rats induced by high-fat diet (HFD) and streptozotocin (STZ) were studied. After oral administration of Mag (25, 50 and 100 mg x kg(-1) x d(-1)) for continuous 10 weeks, the blood glucose and lipids (TC, TG and LDL-C) levels, as well as the hepatic CYP2E1 activity and MDA content of diabetic rats, decreased significantly (P < 0.05 or P < 0.01), whereas the oral glucose tolerance and hepatic antioxidant enzymatic activities (CAT and GSH-Px) of diabetic rats, increased significantly (P < 0.05 or P < 0.01). The results indicated that Mag was effective against the hepatic oxidative damage, hyperglycemia and hyperlipemia of diabetic rats induced by HFD and STZ, and the inhibition of Mag on hepatic CYP2E1 activity could be an important mechanism of Mag against hepatic insulin resistance and oxidative damage.

Objective:To study the effects of ginsenoside Rb1 on cell proliferation,differentiation and OPG/RANKL mRNA expression of rat osteoblast cultured in vitro.Methods:Rat osteoblast cultured in vitro was treated by a series of different concentration of ginsenoside Rb1 for different time respectively.The proliferation of osteoblast was detected by MTT method.The differentiation of osteoblast was detected by the activity of ALP.mRNA expression of osteoprotegerin (OPG) and RANKL(receptor activator nuclear factor-kappa B ligand,RANK) were semi-quantified by RT-PCR,and the ratio of OPG/RANKL mRNA was used to analyze the differentiation of ginsenoside Rb1 to osteoclast.Results:Ginsenoside Rb1 at a concentration of 1.12?10-9mol/L to 1.12?10-5mol/L can promote the proliferation of osteoblast after 72h of treatment (P

Objective:To investigate the effects of Maijun'an Tablets on expression of endothelin-1(ET-1) and nitricoxide synthase(NOS) in tissues and plasma of spontaneous hypertensive rats (SHR). Methods:12 spontaneous hypertensive rats (SHR) were divided randomly into normal control group and drug administration group. Rats in drug administration group were orally given with Maijun'an Tablets (hydrochlorothiazide 10 mg?kg-1.d-1,puerarin 36 mg?kg-1.d-1 and total alkaloid of rhynchophylla 122 mg?kg-1.d-1) once a day for 4 weeks,and rats in control group were orally dosed with equal amount of physiological saline. All animals were sacrificed after 4 weeks. The mRNA and protein contents of ET-1 and NOS in plasma,liver,kidnay and blood vessel were determined. Results:Compared with the control group,Maijun'an Tablets can effectively reduce blood pressure of SHRs,regulate down the content of ET-1 and regulate up the content of NOS at mRNA and protein levels. Conclusion:The up-regulation of NOS and down-regulation of ET-1 at mRNA and protein levels were the major antihypertensive machnisam of Maijun'an Tablets for SHRs.

Objective To investigate the growth curve,breeding rate,and blood physiological and biochemical parameters in Npc1 gene mutant mice (Npc1-/-) for providing theoretical evidence in research on Niemann-Pick disease type C1 (NPC1) patient.Methods 1) The body mass of Npc1-/-,Npc1+/-,and Npc1+/+ mice (n=120;60♀,60♂) was measured from 0 to 77 days;(2) As Npc1-/-mice were born only by the mating Npc1+/-mice,the breeding rate of Npc1+/-mice was counted here from the 1st to 4th generation;(3) The blood physiological and biochemical parameters were measured on both Npc1-/-and Npc1+/+ mice at 60 days.Results 1) Compared with the wild type controls,the body weight of Npc1-/-mice was progressively increased up to 7 weeks and then decreased,and died around 11 weeks.The body weight of the Npc1+/-and Npc1+/+ mice was increased as time went on.After 4 weeks,the male mice showed a higher weight gain than the females;(2) The generations of Npc1+/-mice had no significant difference in mating-parturition interval,litter size,weaning litter and the number of male and female (P>0.05),but the weaning rate of the 2nd generation was significantly higher than that of the 1st generation (P<0.05);(3) The hematological parameters showed a significant difference only in mean corpuscular hemoglobin (MCH) and mean peroxidase index (MPXI) between the Npc1-/-and Npc1+/+ mice (P<0.05).No significant difference was found in other hematological parameters (P>0.05).Among the biochemical parameters,aspartate aminotransferase (AST),glucose (GLU),lactate dehydrogenase (LDH),potassium (K) and copper (Cu) had a significant difference between the Npc1-/-and Npc1+/+ mice (P<0.05).Conclusions 1) The growth curves of Npc1-/-,Npc1+/-,and Npc1+/+ mice are different due to different genotype and sex;(2) The reproduction rates of Npc1+/-mice have no significant difference among different generations;(3) The blood physiological parameters (MCH,MPXI) and biochemical parameters (UREA,AST,GLU,LDH,K,Cu) are significantly different between Npc1-/-and Npc1+/+ mice.

This study sought to determine the impact of dental fluorosis severity on demineralization and remineralization of human fluorosed teeth in vitro. Surface enamel microhardness was measured on the enamel blocks before and after demineralization and after remineralization. The results showed that after demineralization, the sequence of % Surface microhardness demineralization (% SMHD) was TFI4 (18.92 +/- 1.31) < TFI3 (20.50 +/- 1.32) < TFI2 (25.08 +/- 1.69) < TFI1 (27.77 +/- 1.79) < TFI0 (30.70 +/- 1.35) (P < 0.05), and there was no statistically significant differences between TFI1 (27.77 +/- 1.79) and the normal group TFI0 (30.70 +/- 1.35). After remineralization, the sequence of % Surface microhardness remineralization (% SMHR) was TFI1 (55.17 +/- 1.23) > TFI0 (53.97 +/- 3.05) > TFI2 (49.17 +/- 1.81) > TFI3 (44.85 +/- 1.89) > TFI4 (36.51 +/- 2.95) (P < 0.05). Moderately fluorosed enamel showed a significatnt resistance to caries, but mildly fluorosed enamel could get better remineralization. These facts and figures deserve clinicians' attention.
Dental Enamel ; chemistry ; pathology ; Fluorosis, Dental ; metabolism ; Humans ; In Vitro Techniques ; Tooth Demineralization ; Tooth Remineralization

OBJECTIVEDust sample mass gain is too smaller to satisfy the limit of detection (LOD) even in most cases during dust sampling at workplaces nowdays, especially for respirable fraction. Therefore, it is aimed to solve the problem by increasing sample load with high flow rate samplers.

METHODSIn A and B two shipyards respirable welding fume was sampled by high flow rate cyclone samplers of FSP-10 (10 L/min) for 2-2.5 hours and normal flow rate FSP-2 (2 L/min) for 3-4 hours with a stratigy of parallele sampling at the same workpalce, in order to compare their mass gain, coincidence rate with LOD, and airborn dust concentration.

RESULTSSample mass gain of 0.97±0.40 mg and 1.61±0.86 mg respectively in the two factories by FSP-10 was significantly higher than that of 0.29±0.12 mg and 0.51±0.27 mg by FSP-2 (t-test, P<0.05 in both cases) , increasing herewith the coincidence rate with LOD from 26.8% (when sampling with FSP-2, calculated together with samples of the two factories) to 89.7%. However there was no significant difference in dust concentrations by the two different samplers, 0.53±1.88 vs 0.73±1.61 mg/m(3) by FSP-2 and FSP-10 in the shipyard A and 1.14±1.78 vs 1.01±1.63 mg/m(3) in the factory B (t-test, P>0.05 in every case) . In addtion, sample loading by FSP-2 was found to be correlated to sampling time (R(2)=0.7906, y=0.002 6x) , therefore, it has to sample for ≥192.3 min to meet the LOD (0.5 mg) in case of normal flow rate.

CONCLUSIONBy using of high flow rate cyclone FSP-10 the problem of LOD could be solved, along with increased sample mass and similar respirable dust concentration by the two samplers. Some techincal improvements of FSP-10 and increasing of LOD coincidence rate by other methods was also disscussed.

Air Pollutants, Occupational ; analysis ; Construction Industry ; Dust ; analysis ; Environmental Monitoring ; instrumentation ; Occupational Exposure ; Ships ; Workplace

Objective To analyze the differences in immune system between Npc1 gene mutant (Npc1-/ -) and wild-type (Npc1+/ +) mice for better understanding the pathogenesis of Niemann-Pick disease type C1 (NPC1) from an immunological perspective and providing reference for NPC1 treatment in clinic.Methods Body, thymus and spleen weight of Npc1-/ -and Npc1+/ + mice aged (14±2) days, (42±2) days and (63±2) days (Day14±2 , Day42±2 and Day63±2 ) were recorded and the associated organ index were calcu-lated. White blood cell count in peripheral blood of mice aged Day42±2 was examined by routine blood test. Expression of cytokines at mRNA level in mouse peripheral blood was detected by qPCR. Percentages of CD4+, CD8+ and CD19+ lymphocytes in peripheral blood and spleen of mice aged Day42±2 were measured by flow cytometry. Apoptosis and senescence of spleen in mice aged Day63±2 were examined by immunofluores-cence and β-galactosidase staining. Results Compared with Npc1+/ + mice, there was no significant differ-ence in the weight of spleen and thymus in Npc1-/ - mice aged Day14±2; the weight of spleen in Npc1-/ - mice aged Day42±2 significantly increased, but the weight of thymus showed a significant decrease; furthermore, both the weight of spleen and thymus in Npc1-/ - mice aged Day63±2 significantly decreased; and the body weight of Npc1-/ - mice of each age group significantly decreased. Moreover, compared with Npc1+/ + mice, the absolute number of lymphocytes in the peripheral blood of Npc1-/ - mice aged Day42±2 showed no signifi-cant difference, but the percentage in whole white blood cells significantly decreased due to the significantly increased neutrophils. Expression of cytokines ( IL-1, IL-2, IFN-γ, TNF-α, IL-4, granzyme A and granzyme B) at mRNA level in the peripheral blood leukocytes of Npc1-/ - mice aged Day42±2 was abnormal as compared with that in Npc1+/ + mice. The number of T (CD4+ and CD8+) lymphocytes in Npc1-/ - mice aged Day42±2 significantly decreased, while the number of B (CD19+) lymphocytes increased significantly as com-pared with those in the Npc1+/ + mice. Compared with Npc1+/ + mice, apoptosis and senescence of the spleen in Npc1-/ - mice aged Day63±2 aggravated significantly. Conclusion The abnormal lipid metabolism triggered by Npc1 gene mutation causes severe immune dysfunction in Npc1-/ - mice. Therefore, immune dysfunction should be taken into full consideration when treating patients with NPC1, which might help improve the life quality and prolong the survival time.

In recent years, the biopharmaceutical industry has developed rapidly, creating urgent demand for high-quality, innovative, and application-oriented talents. In the context of "first-class undergraduate education", it is of great significance to reform and explore biopharmaceutics blended learning to foster professional talents who can adapt to the industrial development. The blended teaching of biopharmaceutics course in Hubei University was based on small private online course (SPOC) and ChaoXing platform, aiming to meet the first-class "AIC (advanced, innovation, challenge)". The course strengthened the three phases of teaching: before, during, and after class, and innovated teaching methods actively to achieve curriculum goals, and integrated typical cases organically. In addition, the course improved the discriminative power of assessment by strengthening the formative performance evaluation. Moreover, the course provided guidance for students to improve the learning efficiency through investigating the students' learning behavior and employing the marginal utility curve to analyze the characteristics of group activities. Furthermore, the course also offered students personalized learning guidance based on their career planning. The reform of biopharmaceutics blended teaching has achieved significant outcomes, such as improving students' satisfaction, students' innovation and entrepreneurship ability, and curriculum construction level, thus may serve as a reference for the teaching reform and research of the related courses.
Humans ; Biopharmaceutics ; Curriculum ; Learning ; Students

The recombinant adenoviruses expressing miR-22 (Ad-miR-22) was constructed and the effect of Ad-miR-22 on insulin signal pathway and glucose uptake in HepG2 cells was analyzed. MiR-22 gene was amplified by PCR from human hepatocytes and cloned into the pAdTrack-CMV vector to generate the shuttle plasmid pAdT-22. The positive colonies were confirmed by PCR and sequencing. The resultant shuttle plasmid was linearized with Pme I, followed by co-transformation into competent BJ5183 cells containing an adenoviral backbone plasmid (pAdEasy-1) to create the recombinant plasmid pAd-miR-22. After digested with Pac I, the linearized pAd-miR-22 was transfected into 293A packaging cell line to generate recombinant adenoviruses Ad-miR-22. HepG2 cells were infected with Ad-miR-22 or control Ad-GFP (adenoviruses expressing green fluorescent protein), and then the miR-22 expression levels were analyzed by qPCR. The result shows that adenovirus-mediated overexpression of miR-22 significantly decreased insulin-induced glucose uptake in HepG2 cells. Moreover, overexpression of miR-22 markedly decreased insulin-induced phosphorylation of GSK-3β. miR-22 also increased the mRNA levels of gluconeogenic genes in HepG2 cells. Furthermore, Western blotting results indicate that the protein expression of SIRT1 decreased in Ad-miR-22 infected HepG2 cells as compared with Ad-GFP infected HepG2 cells. In summary, overexpressing of miR-22 significantly increased gluconeogenesis while decreased glucose uptake in HepG2 cells. The effect of miR-22 on glucose metabolism may be mediated by SIRT1.
Adenoviridae ; genetics ; Glucose ; metabolism ; Glycogen Synthase Kinase 3 beta ; metabolism ; Hep G2 Cells ; Humans ; MicroRNAs ; genetics ; metabolism ; Signal Transduction ; genetics ; Transfection