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This study aimed to construct an intelligent fluorescent nanocarrier for tumor cell tracing. Doxorubicin (DOX) was used as a model drug, and the gene targeting siBcl-2 was loaded to achieve synergistic inhibition of tumor cells. Mesoporous silicon nanoparticles (MSN) were prepared by a sol-gel method, and acetaldehyde cystine (AC) and polyethyleneimine (PEI) were covalently modified. The prepared nanocarrier MSN-AC-PEI was uniformly dispersed, with a particle size of 235.53 nm and a potential of 14.63 mV. The carrier material MSN-AC-PEI could load siRNA with the mass ratio of 60∶1 (Wvectors∶WsiRNA) and protect siRNA from RNase I degradation. To simulate the microenvironment of tumor, DOX release in phosphate buffer (pH 5) including 10 mmol·L-1 glutathione (GSH) was investigated. The cumulative release rate of DOX at 120 h is 35 times that of the normal physiological environment, which lays the foundation for the intelligent release of DOX in tumor cells. The results of cell experiments showed that the carrier material MSN-AC-PEI had significant green fluorescence, and the traceability can be maintained for 24 h after taken up by MCF-7 cells. After 24 hours of administration of the nano drug delivery system MSN-AC-PEI@DOX/siBcl-2, the inhibition rate of tumor cell proliferation reached 40.91%, and the late apoptosis rate was 60.84%. The Western blot results showed that compared with free DOX and siBcl-2, the nano-delivery system MSN-AC-PEI@DOX/siBcl-2 can significantly reduce the expression of anti-apoptotic protein Bcl-2, thereby enhancing its anti-tumor ability.
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The mortality rate of patients with cardiac arrest remains very high. At present, many unknown areas on cardiopulmonary resuscitation science need to be further investigated so as to provide scientific basis for the update of international guidelines of cardiopulmonary resuscitation and provide new ideas and new treatment means for improving the prognosis of patients. The standardization of research subjects is vital for any researches on cardiopulmonary resuscitation. From the standardized collection of data of patients with cardiac arrest, to the highly biomimetic manikins, and the construction of animal and cell models, each has its own advantages and disadvantages. Researchers can choose the appropriate research model according to their own experimental purposes.
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To study the effects of saikosaponin b2( SS-b2) on inflammatory factors and energy metabolism against lipopolysaccharide/galactosamine( LPS/Gal N) induced acute liver injury in mice. Mice were randomly divided into normal group( equal amount of normal saline),model group( 100 g·kg~(-1) LPS and 400 mg·kg~(-1) Gal N),low,medium,high dose group of SS-b2( SS-b25,10,20 mg·kg~(-1)·d-1) and positive control group( dexamethasone,10 mg·kg~(-1)). All of the groups except for the normal group were treated with LPS/Gal N though intraperitoneally injection to establish the acute liver injury model. The organ indexes were calculated. The levels of serum transaminases( ALT and AST) and the activities of ATPase( Na+-K+-ATPase,Ca2+-Mg2+-ATPase) in liver were detected. The activity of tumor necrosis factor-α( TNF-α),interleukin-1β( IL-1β) and interleukin-6( IL-6) were determined by the enzyme-linked immunosorbent assay( ELISA). The contents of lactate dehydrogenase( LDH) in liver were determined by micro-enzyme method. HE staining was used to observe the histopathological changes of the liver. Histochemical method was used to investigate the protein expression of liver lactate dehydrogenase-A( LDH-A). The protein expressions of Sirt-6 and NF-κB in the liver were detected by Western blot. According to the results,compared with the model group,there were significant changes in organ indexes in the high-dose group of SS-b2( P<0. 05). The level of ALT,AST,TNF-α,IL-1β,IL-6 and the activities of LDH in serum of mice with liver injury were significantly reduced in the medium and high dose groups of SS-b2( P<0. 01). With the increase of the concentration of SS-b2,the range of hepatic lesions and the damage in mice decreased. The activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase in liver of mice were significantly enhanced in each dose group( P<0. 01). The expression of NF-κB in liver tissues was significantly down-regulated in the medium and high dose group( P<0. 01). Meanwhile,the expression of Sirt-6 protein in the liver of mice with acute liver injury was significantly increased in each dose group( P<0. 01).In summary,SS-b2 has a significant protective effect on LPS/Gal N-induced acute liver injury in mice,which may be related to the down-regulation of NF-κB protein expression and up-regulation of Sirt-6 protein expression to improve inflammatory injury and energy metabolism.
Subject(s)
Animals , Mice , Chemical and Drug Induced Liver Injury , Drug Therapy , Cytokines , Metabolism , Energy Metabolism , Galactosamine , Inflammation , Drug Therapy , Lipopolysaccharides , Liver , NF-kappa B , Metabolism , Oleanolic Acid , Pharmacology , Random Allocation , Saponins , Pharmacology , Sirtuins , MetabolismABSTRACT
Objective To investigate the mechanism of Toll-like receptor in intestinal mucosal injury induced by Cryptospo-ridium parvum infection in mice.Methods Totally 30 male BALB/c mice were randomly divided into a normal control group,1-week infection group and 2-week infection group.The mice of the 1-week and 2-week infection groups were sacrificed 7 days and 14 days after the infection respectively,and the mice of the normal control group were sacrificed 14 days after the infection.The model of intestinal infection of C.parvum in mice was built by using the immunosuppressive method and oocyst intragastric ad-ministration.The pathological changes of the intestinal mucosa of mice were observed with a light microscope and the villus height,crypt depth and ratio of villus height/crypt depth were measured.The ultrastructure of the intestinal mucosa of mice was observed by a transmission electron microscope(TEM).The expressions of TLR2 and TLR4 in the intestinal mucosa were tested by qPCR and Western blotting.Results Under the light microscope,the intestinal villi were dropsical,obviously atrophied and shortened,and the submucosal structure was dropsical.The height of chorionic villi and the ratio of villus height to crypt depth in the jejunum of the 1-week and 2-week infection groups were significantly lower than those in the normal control group(all P<0.05),while the depth of the recess of the former two was significantly increased(all P<0.05).With the extension of the infection time,the villus height and the ratio of villus height to crypt depth in the jejunum of mice decreased significantly(both P<0.05),and the crypt depth increased significantly(P<0.01).The TEM observation showed that the structure of the oocyst of C.parvum in the jejunum of the infected mouse was intact,the villi around the oocyst were abscission seriously,and the oocyst wall was fused with the epithelial cell membrane.The qPCR observation showed that compared with the normal control group,the expressions of TLR2 mRNA and TLR4 mRNA in the intestinal mucosa of the 1-week and 2-week infection groups were significantly higher(all P<0.05).In addition,the expressions of TLR2 and TLR4 mRNA in the 2-week infection group were significantly higher than those in the 1-week infection group(both P<0.05).The Western blotting showed that the expres-sions of TLR2 protein and TLR4 protein in the intestinal mucosa of the 1-week and 2-week infection groups were significantly higher than those of the normal control group(all P<0.05).Furthermore,the expressions of TLR2 and TLR4 protein in the 2-week infection group were significantly higher than those in the 1-week infection group(both P<0.05).Conclusions TLR2 and TLR4 are important receptors for intestinal mucosal recognition of C.parvum.The C.parvum infection may lead to intestinal mucosal damage possibly via the mechanisms associated with the up-regulation of TLR2 and TLR4 expressions.
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Objective To evaluate and compare the primary clinical efficacy in high myopia patients undergoing implantable collamer lens (ICL) implantation or femtosecond laser-LASIK (FS-LASIK).Methods Totally 73 patients with myopia from August 2016 to March 2017 in our hospital were collected and divided into ICL group(35 patients with 69 eyes) who receiving ICL implantation and LASIK group (38 patients with 76 eyes) who underwent FS-LASIK.There was no significant difference in gender composition and age between the two groups (both P ＞ 0.05).In addition,there was no significant difference in contrast sensitivity function (CSF) and glare CSF (GCSF) between the two groups before operation (both P ＞ 0.05).After 6 months of follow-up,the patient's uncorrected visual acuity,best corrected visual acuity and equivalent spherical degree were detected,and efficacy index and safety index were calculated.Meanwhile,CSF and GCSF examination were performed,followed by the observation of the occurrence of complications.Results There was no significant difference in the uncorrected visual acuity and best corrected visual acuity between the two groups before surgery (all P ＞ 0.05).The preoperative spherical equivalent in the ICL group was -4.000 to-12.00 (-8.86 ±3.70)D,and-4.00 to-11.75 (-8.51 ±4.20)D in the LASIK group,with no statistically significant difference (P ＞ 0.05).There was no significant difference in the uncorrected visual acuity,best corrected visual acuity and spherical equivalent between the two groups after surgery (all P ＞ 0.05).The efficacy coefficient and safety index of the ICL group were better than those in the LASIK group after 6 months,and the differences were statistically significant (all P ＜ 0.05).The CSF in the ICL group was at 1.5 c · d-1,3.0 c · d-1,6.0 c · d 1,12.0 c · d-1,and 18.0 c · d-1 after surgery,which was superior to the LASIK group,and the differences were statistically significant (all P ＜ 0.05).The GCSF in the ICL group was at 1.5 c · d-1,3.0 c · d-1,6.0 c · d-1 and 18.0 c · d-1 after surgery,which was superior to the LASIK group,and the differences were statistically significant (all P ＜ 0.05).Four hours after operation,there were 4 patients (7 eyes) with increased intraocular pressure,which were controlled by anterior chamber puncture,but no lens opacity presented.Conclusion Both ICL and FS-LASIK can effectively correct moderateto highmyopia,but ICL is better than FS-LASIK at the effectiveness index,the safety index,CSF and GCSF.
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AIM:To evaluate the clinical effects of femto-LASIK using new Ziemer LDV Z6 femtosecond laser machine (Z6). METHODS: Two - hundred cases ( 400 eyes ) was randomly separated into two groups: Group A included 200 eyes which corneal flaps were made by Z6, and Group B included rest of 200 eyes which corneal flaps were examined by a traditional Ziemer LDV CrystalLine femtosecond laser machine (CrystalLine). Visual acuity, slit lamp, refraction, Sim-K average, intraocular pressure (IOP), non-invasive tear break-up time (NIAvg-BUT), operation difficulty and complications were compared between two groups preoperatively and postoperatively.RESULTS:There was no significant differences between two groups in visual acuity, refraction, Sim-K average, IOP and NIAvg - BUT either preoperatively or 6mo postoperatively (P>0. 05). Although there were significant differences in operation difficulty and complications were found between two groups ( P CONCLUSION:More careful and strict requirements are needed when using the new Z6 femtosecond laser for corneal flaps.
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?AIM:To evaluate the objective visual quality of patients who underwent corneal cross-linking for the keratoconus using double-pass analysis system.? METHODS: Advanced keratoconus patients who underwent UV - riboflavin corneal cross - linking from January to July 2015 were included. The outcomes of their objective scattering index ( OSI ) , predicted visual acuity ( VA ) , the cut - off frequency of modulation transfer function ( MTF cut- off ) , the Strehl ratio ( SR ) were compared before and 6mo after corneal cross-linking.?RESULTS: A total of 13 patients ( 16 eyes ) were included. There was no statistically significant difference between pre- and 6mo postoperative data in uncorrected visual acuity, best corrected visual acuity, refractions and mean value of Sim-k (P>0. 05). Non-invasive average tear film break up time ( NIAvg-BUT ) detected by the Sirius system decreased after corneal cross-linking ( P0. 05). Tear Film Analysis Mean OSI increased at 6mo postoperatively (P<0. 05).? CONCLUSION: The subjective visual quality isn’t effected by corneal cross-linking. The tear stabilities of patients are influenced by these operations at 6mo postoperatively. More observations on long-term effect are needed to be taken in the future.
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<p><b>OBJECTIVE</b>To study the relationship between JWA polymorphisms and the susceptibility to hypertension in workers exposed to heat stress.</p><p><b>METHODS</b>The exposure group included 158 steelworkers and rollers and 76 workers unexposed to heat stress served as control group. The general information was collected according to a questionnaire and the blood pressure was examined for all subjects. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to analyze the site 76 in promoter and site 723 in the 3rd exon of JWA gene in the peripheral lymphocytes. PHASE 2.0 software was utilized to calculate the haploid type.</p><p><b>RESULTS</b>In the exposure group, JWA76 G/G genotype frequencies of sub-group with normal blood pressure, sub-group with higher blood pressure and sub-group with hypertension were 82.35%, 69.70% and 65.00%, respectively, there were significant differences among 3 sub-groups (P < 0.05). JWA 76 G/G genotype frequencies decreased with blood pressure (χ² = 4.86, P = 0.027). The multiple logistic regression analysis showed that the subjects with G/C genotype were compared to the subjects with G/G genotype in the exposure group, the adjusted OR value was 3.67 (95%CI: 1.21 approximately 11.05) for the risk of hypertension and higher blood pressure. the subjects with GG/CT haploid type were compared to the subjects with non-GG/CT haploid type in the exposure group, the adjusted OR values for the risks of hypertension and higher blood pressure were 8.30 (1.39 approximately 49.44) and 8.55 (1.53 approximately 47.48), respectively.</p><p><b>CONCLUSION</b>The gene polymorphisms at site 76 and GG/CT haploid type of JWA gene were associated with hypertension in workers exposed to high temperature.</p>
Subject(s)
Adult , Humans , Male , Blood Pressure , Genetics , Control Groups , Gene Frequency , Genetic Predisposition to Disease , Genotype , Heat-Shock Proteins , Genetics , Hot Temperature , Hypertension , Epidemiology , Genetics , Intracellular Signaling Peptides and Proteins , Genetics , Polymorphism, Single Nucleotide , WorkplaceABSTRACT
The objective of this study was to investigate the protection by naringenin against doxorubicin-induced oxidative damage in normal blood cells. Inhibiting effects of naringenin, doxorubicin and naringenin combined with doxorubicind on K562 cells and polymorphonuclear leukocytes were detected with MTT method, the level of reactive oxygen species (ROS) and lipid peroxidation (MDA), the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were examined with spectrophotometric method in the K562 cells and polymorphonuclear leukocytes. The results indicated that the proliferation of K562 cells was not inhibited by the cytotoxicity of doxorubicin in combination of naringenin with doxorubicin. As compared with the doxorubicin, the addition of naringenin after doxorubicin for 1 hour, the levels of reactive oxygen species (ROS) and lipid peroxidation (MDA) obviously decreased, the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) obviously increased in the polymorphonuclear leukocytes, but these were not changed obviously in K562 cells. It is concluded naringenin can protect against doxorubicin-induced oxidative damage in normal blood cells. The mechanism of naringenin may be elevating activities of antioxidant enzyme and degrading oxidative production level in normal blood cells, and meanswhile decreasing level of oxidative products.
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Humans , Antioxidants , Pharmacology , Doxorubicin , Erythrocytes , Flavanones , Pharmacology , Oxidative Stress , Reactive Oxygen Species , Metabolism , Superoxide Dismutase , MetabolismABSTRACT
The objective of this study was to investigate the apoptosis-inducing effect and underlying mechanism of naringenin (NGEN) on K562 cells in vitro. The inhibition of NGEN on K562 cells was evaluated by means of MTT assay so as to observe the cytotoxicity of NGEN; The morphological changes of the cells treated by NGEN were observed by transmission electron microscope; cell apoptosis rate influenced by NGEN was assessed by flow cytometry; the enzyme activity changes of caspase-3 and caspase-8 in the process of NGEN-induced K562 apoptosis were detected by Caspase Colorimetric Assay Kit; immunohistochemistry technique was used to detect FAS, FASL protein expression in K562 cells. The results showed that the growth of cells was inhibited by NGEN in dose-and time-dependent manners (p<0.05); NGEN-induced K562 cells apoptosis and sub-G1 peak was observed; some typically early and final phase changes of cell apoptosis were revealed under transmission electron microscope; the enzyme activity of caspase-3 and caspase-8 and the expression of FAS remarkably increased, meanwhile the expression of FASL was down-regulated (p<0.05). It is concluded that NGEN exerts anti-cancer effect by inducing K562 cell apoptosis, and the regulation of the expression of FAS and FASL. The caspase-3 and caspase-8 co-pathway brings about one of the mechanisms.
Subject(s)
Humans , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Caspase 3 , Metabolism , Caspase 8 , Metabolism , Fas Ligand Protein , Genetics , Metabolism , Flavanones , Pharmacology , K562 Cells , fas Receptor , Genetics , MetabolismABSTRACT
<p><b>AIM</b>To investigate the effect of oridonin (ORI) on telomerase activity and cell cycle of human leukemic cell line K562 cells.</p><p><b>METHODS</b>Immunohistochemistry (IHC) technique was used to determine the expression of hTERT or C-myc. Telomerase activity was detected with TRAP-PCR-ELISA assay. In addition, the percentages of K562 cells in different cell cycle were determined by flow cytometry (FCM) at 24th and 48th hours separately after adding the different concentrations of ORI.</p><p><b>RESULTS</b>After the K562 cells were treated with ORI at 3.43 micromol x L(-1) for 48 h, the expression of hTERT and C-myc decreased obviously. There was statistical significant (P < 0.05) difference between experimental groups and the normal controls. In addition, the telomerase activity of K562 cells was significantly inhibited by ORI at the dose of 3.43 micromol x L(-1) for 48 h. At the same time, the cell cycle distribution changed, the percentage of G0/G1 or G2/M stages cells increased and that of the S stage cells decreased after ORI was added.</p><p><b>CONCLUSION</b>ORI can effectively inhibit telomerase activity in K562 cells. Arresting cell cycle and decreasing the expression of hTERT and C-myc may be the mechanism of action.</p>
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Humans , Antineoplastic Agents, Phytogenic , Pharmacology , Cell Cycle , DNA-Binding Proteins , Diterpenes , Pharmacology , Diterpenes, Kaurane , Isodon , Chemistry , K562 Cells , Plants, Medicinal , Chemistry , Proto-Oncogene Proteins c-myc , Metabolism , Telomerase , MetabolismABSTRACT
Chromogranin A (CGA) is a soluble protein existed in most secreted cells and neurons. It was recently found that the bovine CGA N terminal region has vasoinhibitory, antibacterial and antifungal activities. Since the need for effective antifungal agents increases in parallel with the expanding number of immunocompromised patients at risk for fungal infections, it becomes imperative to find antifungal compounds with low toxicity toward mammalian cells. To study the antifungal activity of CGA N terminal region, the DNA fragment encoding for the N terminal 1-76 amino acid sequence (CGA1-76) of human CGA was amplified by PCR technique. After DNA sequence analysis, the amplified DNA fragment was cloned into the Bacillus subtilis inducible and expression vector pSBPTQ constructed in this study and the resultant plasmid pSVTQ was then transformed into triple-protease deficient Bacillus subtilis strain DB403 competent cells. The transformants was screened on LB plates containing 10 microg/mL kanamycin. The positive transformant DB403 (pSVTQ) was grown on kanmycin containing 2 x MSR medium and sucrose was added to 2% final concentration for induction after 2h cultivation. The culture supernatant was used to run SDS-PAGE. The result of SDS-PAGE showed that the CGA1-76 was expressed by sucrose induction and the expressed product secreted into the medium with a yield of 5 mg/L. The expressed product reacts specifically with mouse anti CGA47-68 monoclonal antibody. The antifungal activity of the expressed product was examined by adding the culture supernatant to the fungal spore or Candida albican suspensions at appropriate proportion and found that the recombinant human CGA1-76 produced in Bacillus subtilis inhibits the growth of Fusarium sp. Alternaria sp. and Candida albican at the concerntration of 4 micromol/L. These results demonstrate that human CGA1-76 has expressed in Bacillus subtilis and the expressed product is immunogenic and has the antifungal activity.