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Background Acute exposure to mercury chloride (HgCl2) can cause liver damage. Whether oleanolic acid (OA) as a hepatoprotective drug can protect against liver injury induced by acute exposure to HgCl2 and related mechanism of action remain unclear. Objective To investigate the protective effect and possible mechanism of OA on liver injury in mice caused by acute exposure to HgCl2. Methods Forty SPF C57BL/6 male mice were randomly divided into four groups with 10 mice in each group according to body weight. The four groups were named control group, OA group (300 mg·kg−1), HgCl2 group (5 mg·kg−1), and OA + HgCl2 group (300 mg·kg−1 OA + 5mg·kg−1 Hgcl2). Soybean oil and OA solution were administered intragastric once a day for two consecutive days. HgCl2 solution was injected intraperitoneally 2 h after the second intragastric administration. Mice were sacrificed after 48 h, and their serum and liver were collected. Liver coefficient was calculated. The changes of liver structure and iron deposition were observed by hematoxylin-eosin (HE) staining and Prussian blue staining. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), total superoxide dismutase (T-SOD), reduced glutathione (GSH), malondialdehyde (MDA), and tissue iron content were measured with commercial kits. Western blotting was used to detect nuclear factor erythroid-2 related factor 2 (Nrf2), heme oxygenase 1 (HO-1), glutathione peroxidase 4 (Gpx4), transferrin receptor 1 (TFR1,) and solute carrier family 7 member 11 (SLC7A11). Results The AST and ALT levels of the HgCl2 group were (76.447±9.695) U·g−1 and (98.563±24.673)U·g−1, respectively, which were higher than those of the control group (P<0.05). After the OA pretreatment, the liver coefficient and the above indexes were decreased to (4.769±0.237)%, (57.086±10.087) U·g−1, and (87.294±27.181)U·g−1, respectively. The liver coefficient and AST level of the OA + HgCl2 group were significantly different from those of the HgCl2 group (P<0.05). After acute exposure to HgCl2, the hepatocytes of mice were disordered, accompanied by inflammatory infiltration, positive blue particles appeared in Prussian blue staining of liver tissue, and the above changes in liver tissue were alleviated after the OA pretreatment. The iron content in the HgCl2 group was (3.646±0.238) μmol·g−1, which was higher than that in the control group, (2.948±0.308) μmol·g−1. After the OA pretreatment, the iron content decreased to (3.429±0.415) μmol·g−1. Compared with the control group, acute exposure to HgCl2 resulted in decreased levels of GSH and T-SOD, decreased protein expression levels of Nrf2, HO-1, SLC7A11, and Gpx4, increased level of MDA, and increased protein expression level of TFR1 (P<0.05). After the OA pretreatment, all indicators were improved including increased GSH level, decreased MDA level, increased Nrf2, HO-1, and SLC7A11 protein expression levels, and decreased TFR1 protein expression level; compared with the HgCl2 group, the differences were statistically significant (P<0.05). Conclusion Acute HgCl2 exposure could induce liver injury in mice, and its mechanism may involve iron overload and ferroptosis. OA may alleviate the liver injury caused by acute HgCl2 exposure by affecting iron overload and the ferroptosis-related protein expression.
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Based on the International Federation of Nursing Anesthetists (IFNA) education and training base, our hospital has built a training base for anesthesia specialized nurses in Nanjing from the following aspects: the application for training base of anesthesia specialized nurses, the qualification and examination of students' and teachers' qualification, the settings of training curriculum, the examination contents and methods, and the evaluation of post-training effect. This article summarizes the construction experience of this base, therefore, providing support and standard for the training of anesthesia specialized nurses.
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OBJECTIVE：To investigate the correlat ion betwe en N-methyl-D-aspartate（NMDA）receptor subunit gene GRIN2B polymorphism and Han population with epilepsy in south Fujian. METHODS ：In retrospective study ，167 healthy people who had physical examination in the Affiliated Dongnan Hospital of Xiamen University were selected from Jan. 2017 to May 2018 as control group；163 epileptic patients who were monitored the blood concentration of sodium valproate were selected as epilepsy group. The clinical data and peripheral blood of 2 groups were collected. 12 loci of GRIN2B genotype（rs11055514，rs11055515，rs12814951， rs74816802，rs2160517，rs2193149，rs966664，rs1805476，rs1806201，rs1805522，rs3764030，rs1019385） in subjects were genotyped. Haploview 4.2 software was used to perform linkage disequilibrium （LD）analysis，and Pearson correlation was used to analyze haplotype. Distribution differences of wild homozygote （AA），mutant heterozygote （Aa）and mutant homozygote （aa） genotypes at 12 loci of GRIN2B gene between 2 groups were analyzed statistically by using GENO ，TREND，DOM and REC. Logistic regression model was used to analyze the correlation of epilepsy induction among 12 loci of GRIN2B gene. RESULTS ： Totally 12 loci of GRIN2B gene were all in line with Hardy-Weinberg equilibrium in 2 groups（P＞0.05）. There was an obvious LD phenomenon between the block 1 composed of rs 11055514，rs11055515，rs12814951，rs74816802，rs2160517，rs2193149 and rs966664 and the block 2 composed of rs 3764030 and rs 1019385（D’＞0.9，r2＞1/3）. There was a correlation between CGGACAG monoploid in block 1 and the occurrence of epilepsy （P＜0.05）. There was statistical significance in the distribution difference of rs74816802 and rs 2193149 between 2 groups（P＜0.05）. The mutation of rs 2193149 locus may cause epilepsy （addition and effect of alleles ：OR＝1.529，L95＝1.017，P＝0.041）. CONCLUSIONS ：The mutation of GRIN2B gene rs 2193149 locus may be one of the risk factors of epilepsy in Han population from south Fujian.
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Objective: To deeply investigate the effect of S100A6 in the microenvironment on migration of colorectal carcinoma LoVo cells, and to explore the possible molecular mechanism. Methods: The recombinant protein glutathione S-transferase (GST) and the fusion protein GST-human S100A6 (GST-hS1 00A6) were prepared and identifed. After the treatment with GST-hS1 00A6 (30 (μg/mL), the migration of LoVo cells was detected by wound healing assay, the expressions of total protein kinase B (PKB, also known as Akt) and phosphorylation of Akt (p-Akt) in LoVo cells were detected by Western blotting. After the treatment with GST-hS1 00A6 and phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway inhibitor LY294002 alone or in combination, the migration of LoVo cells was detected by wound healing assay. After the macrophages were treated with the conditioned medium of LoVo cells stimulated by GST-hS1 00A6 (named as CM-A6-LoVo), the expression levels of M2 phenotype marker CD206 and M1 phenotype marker inducible nitric oxide synthase (iNOS) in the macrophages were detected by real-time fuorescent quantitative PCR, and the migration ability of LoVo cells co-cultured with the macrophages was detected by wound healing assay. Results: GST-hS100A6 and GST protein (as a control) were successfully prepared. Compared with GST group, the wound healing rate of LoVo cells treated with GST-hS1 00A6 was significantly elevated (P 0.05), and the macrophages co-cultured with LoVo cells could significantly promote the migration of LoVo cells (P < 0.05). Conclusion: S100A6 in microenvironment can directly and indirectly promote the migration of colorectal carcinoma LoVo cells, which maybe related to the regulation of PI3K/Akt signaling pathway and the induction of macrophage polarization to M2 phenotype.
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AIM:To detect the endogenous expression of B-cell leukemia/lymphoma 6 member B (BCL6B) in FHC and LoVo cells, and to investigate the effects of BCL6B on proliferation and migration of LoVo cells for further explo-ring the underlying mechanism .METHODS:The endogenous expression of BCL 6B in the FHC and LoVo cells was detec-ted by RT-PCR and Western blot .The methods of MTT assay , colony formation assay , wound healing assay and Transwell chamber experiment were employed to examine the biological functions of BCL 6B in the LoVo cells.The mRNA and protein levels of BCL6B, cyclin D1 and matrix metalloproteinase-9 ( MMP-9) were determined by RT-PCR and Western blot , re-spectively.The level of phosphorylated protein kinase B (p-AKT) was detected by Western blot.RESULTS:BCL6B ex-pression was notably repressed in the LoVo cells as compared with the FHC cells , which were significantly increased by transfection with pcDNA3.1-BCL6B.The abilities of proliferation and migration of the LoVo cells at 72 h were inhibited by 28.33%(P<0.01) and 36.11%(P<0.05) in BCL6B group.The mRNA levels of cyclin D1 and MMP-9 in the cells of BCL6B group were decreased by 39.90%(P<0.01) and 77.36% (P <0.05), and the protein levels of cyclin D1, MMP-9 and p-AKT were reduced by 44.00%(P<0.05), 47.06%(P<0.01) and 32.88% (P<0.05), respectively. CONCLUSION:BCL6B inhibits proliferation and migration of the LoVo cells , and the PI3K/AKT signaling pathway is in-volved in this process .
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BACKGROUND:The research and development of scaffold materials is the key to tissue engineering, as the scaffold can provide a stable external environment for cell growth.OBJECTIVE:To summarize the clinical advances in tendon tissue engineering materials.METHODS:We searched CBM, CNKI, CSTJ and PubMed database for relevant articles published from January 2004 to May 2016. The keywords were tissue engineering, tendon injuries, biological scaffold, tendon healing in Chinese and English, respectively.RESULTS AND CONCLUSION:The commonly used tissue engineering tendon materials include natural polymer materials, biological derivatives, synthetic materials and composite materials. Natural polymer materials retain the three-dimensional network structure of the normal tissue, with good biocompatibility but poor mechanical properties and degradation speed. Synthetic polymer materials present with good mechanical properties and biodegradability, but have low hydrophilicity and poor cell adhesion capability. Composite materials as an effective combination of the two above-mentioned materials exhibit a certain potential in clinical practice. Biological derivatives come from organisms, and have a net structure and biomechanical properties most similar to the human body after appropriate treatment. Additionaly, these derivatives also have the normal physiological activity and functions, which are considered as the future development direction of biomedical materials.
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Objective To investigate the effect of internal fixation with proximal femoral nail antirotation (PFNA) and bone cement on peritrochanteric tumors due to metastatic carcinoma.Methods Clinical data of 19 patients with peritrochanteric tumors due to metastatic carcinoma during June 2007 and July 2013 treated with PFNA and bone cement were retrospectively analyzed.Results Visual analogue scale(VAS) was (8.37 ± 1.12) scores before surgery,and (2.58 ± 1.26) scores after 3 d surgery.There was significant difference (t =22.45,P ＜ 0.05).Conclusion For patients with peritrochanteric tumors due to metastatic carcinoma,internal fixation with PFNA and bone cement is an effective way to relieve pain,restore and improve hmb function and enhance quality of life.
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Objective Nano-structured lipid carriers ( NLC) is a concerned research area in pharmaceutics , which as a novel drug delivery system to promote oral absorption of poorly soluble drugs .The literatures were reviewed to explore and summarize the mechanism that nano-structured lipid carriers in promotion of the absorption of poorly soluble drugs .
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Objective To review the newly developed solid self-microemulsifying drug delivery system (S-SMEDDS).Meth-ods Relevant literatures at home and abroad in recent years were consulted and summarized .Results Solid self-microemulsions car-rier, solidification technology and controlled release formulations were discussed , in order to provide relevant references for improving the bioavailability of water-insoluble drugs and the SMEDDS technology for drug release characteristics .Conclusion The utilization of the solid self-emulsifying drug delivery system could significantly enhance the oral bioavailability of water -insoluble drugs .As a new formulation, S-SMEDDS presented huge potential .
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Objective To prepare insulin thiolated hyaluronic acid nanoparticles (Ins-HA-Cys-NPs)and study its physicochemical properties. Methods The Ins-HA-Cys-NPs was prepared by ultrasonic emulsifying method,and the properties of nanoparticles including morphology,mean diameter,Zeta potential,entrapment efficiency and drug loading efficiency were studied,as well as the cryoprotectant selection.Results The prepared nanoparticles was round in appearance and the mean diameter was(178.5 ±0.8)nm,the polydispersity index was (0.214 ±0.013)and the Zeta potential was -(38.47 ±0.46 )mV,while the entrapment efficiency was (48.85 ±0.66 )%,drug loading efficiency was (4.79 ±0.13 )%;10%mannitol as cryoprotectant provided uniform and well dispersed suspension of nanoparticles with blue opalescence after redispersion.Conclusion The thiolated hyaluronic acid nanoparticles may be used as the carrier for oral drug delivery system of insulin,and it provides a basis for studies on rats in vivo.
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BACKGROUND:Recently,researches have found that insulin-like growth factor-1(IGF-1)can induce the differentiation of bone marrow-derived mesenchymal stem cells(BMSCs)into chondrocytes,but there are no reports concerning the differentiation of adipose-derived mesenchymal stem cells(ADMSCs)into chondrocytes induced by IGF-1,as well as interaction with transforming growth factor-β1(TGF-β1)during this process.OBJECTIVE:To explore the possibility of inducing ADMSCs chondrogenic differentiation by using IGF-1 and the interaction with TGF-β1 in induction.METHODS:ADMSCs were obtained,and seeded at 2×10~5 cells/cm~2 in culture flask.Insulin-free chondrogenic media containing IGF-1 or(and)TGF-β1 were used to induce ADMSCs.2 weeks later,cells were harvested and stained by using toluidine blue and collagen Ⅱ antibody immunohistochemistry.Intracellular sulfated proteoglycan and collagen Ⅱ coloring were observed.Reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the expression of collagen Ⅱ,aggrecan and Sox9 mRNA.RESULTS AND CONCLUSION:After induced,toluidine blue stain exhibited that the cells in the three induction groups were polygonal,with cytoplasm and cell membrane of blue different dyeing.Immunohistochemistry for type Ⅱ collagen demonstrated that cytoplasm and cell membrane were stained brown in three induction groups.RT-PCR revealed that the expression of collagen Ⅱ,aggrecan,Sox9 mRNA of IGF + TGF group were significantly greater than the IGF and TGF groups,and IGF and TGF groups were significantly stronger than the control group.No significant difference was determined between the IGF and TGF groups.These results indicated that IGF-1 can induce chondrogenic differentiation from ADMSCs,expressing chondrocyte specific cell phenotype.There is synergism of IGF-1 and TGF-01 to induce the differentiation of ADMSCs into chondrocytes.
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OBJECTIVE: To establish headspace capillary gas chromatography for the content determination of 3 kinds of residual organic solvent (furanidine,methanol and ethanol) in betamethasone sodium phosphate raw material.METHODS: The sample was dissolved in water and n-propanol was used as internal standard.The residual solvent in betamethasone sodium phosphate was separated on HP-INNOWAX (PEG) capillary column with column temperature set at 60 ℃.The injector temperature and FID detector temperature were controlled at 180 ℃ and 250 ℃,respectively.The carried gas was nitrogen at flow rate of 1.0 mL?min-1.The splitting-ratio was 10 ∶ 1.The containers of head-space injector were in equilibrium at 80 ℃ for 30 min.Injection time was 1 min.RESULTS: With this chromatographic condition,those solvents could be separated completely.The linear range were 0.014 4～0.071 8 mg?mL-1 for furanidine,0.060～0.300 mg?mL-1 for methanol and 0.099 3～0.497 mg?mL-1 for ethanol.The average recovery were 103.7% (RSD=0.53%,n=6),95.8% (RSD=0.30%,n=6) and 95.0% (RSD=0.48%,n=6) respectively.The minimum quantitation limit were 0.057 3 ?g?mL-1,0.486 ?g?mL-1,0.145 ?g?mL-1,respectively.3 kinds of residual organic solvents were all in line with the standard stated in Chinese Pharmacopeia.CONCLUSION: The established method is simple,sensitive and accurate for the content determination of residual solvents in betamethasone sodium phosphate raw material.