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1.
Biomedical and Environmental Sciences ; (12): 419-436, 2022.
Article in English | WPRIM | ID: wpr-927681

ABSTRACT

Objective@#To investigate the function of primary cilia in regulating the cellular response to temozolomide (TMZ) and ionizing radiation (IR) in glioblastoma (GBM).@*Methods@#GBM cells were treated with TMZ or X-ray/carbon ion. The primary cilia were examined by immunostaining with Arl13b and γ-tubulin, and the cellular resistance ability was measured by cell viability assay or survival fraction assay. Combining with cilia ablation by IFT88 depletion or chloral hydrate and induction by lithium chloride, the autophagy was measured by acridine orange staining assay. The DNA damage repair ability was estimated by the kinetic curve of γH2AX foci, and the DNA-dependent protein kinase (DNA-PK) activation was detected by immunostaining assay.@*Results@#Primary cilia were frequently preserved in GBM, and the induction of ciliogenesis decreased cell proliferation. TMZ and IR promoted ciliogenesis in dose- and time-dependent manners, and the suppression of ciliogenesis significantly enhanced the cellular sensitivity to TMZ and IR. The inhibition of ciliogenesis elevated the lethal effects of TMZ and IR via the impairment of autophagy and DNA damage repair. The interference of ciliogenesis reduced DNA-PK activation, and the knockdown of DNA-PK led to cilium formation and elongation.@*Conclusion@#Primary cilia play a vital role in regulating the cellular sensitivity to TMZ and IR in GBM cells through mediating autophagy and DNA damage repair.


Subject(s)
Humans , Antineoplastic Agents, Alkylating/therapeutic use , Brain Neoplasms/metabolism , Cell Line, Tumor , DNA/therapeutic use , Glioblastoma/metabolism , Radiation, Ionizing , Temozolomide/therapeutic use
2.
Chinese Journal of Applied Physiology ; (6): 339-345, 2019.
Article in Chinese | WPRIM | ID: wpr-776501

ABSTRACT

OBJECTIVE@#To understand and analyze the rules of endurance exercise on the cerebral cortex adaptive mechanism in aged rats.@*METHODS@#In this study, 3-month-old (n=20), 13-month-old (n=24) and 23-month-old (n=24) specific-pathogen free (SPF) male Sprague-Dawley Rat (SD) rats were divided into young (Y-SED), middle-aged (M-SED) and old-aged (O-SED) sedentary control group, and the corresponding Y-EX, M-EX and O-EX in the endurance exercise runner group. The 10-weeks of regular moderate-intensity aerobic exercise intervention were carried out in the endurance exercise runner group. The exercise mode is treadmill exercise (slope 0), and the exercise intensity gradually increases from 60%~65% of the maximum oxygen consumption (V·O) to 70%~75%, and the exercise time is 10 weeks. Hematoxylin and eosin (HE) staining was used to detect age-related morphological changes. The expressions of superoxide dismutase(SOD) and brain-derived neurotrophic factor (BDNF) and the expressions of synapsin 1 (SYN1) and Ca/calmodulin- dependent protein kinases IIα (CaMK IIα) / AMP-activated protein kinase α1(AMPKα1) / mammalian target of rapamycin (mTOR) pathway -related genes were detected.@*RESULTS@#The cerebral cortex structure of the rats in each group showed age-related aging changes, the expression of SOD in the cortex showed a gradual decline, the expression of BDNF showed an age-increasing trend, and the expression levels of SYN1 and CaMK IIα were increased with age. The changes in AMPKα1 and SirT2 and IP3R, AKT1 and mTOR mRNA levels were increased slightly in middle-aged rats and decreased in aged rats. Compared with the rats in each sedentary control group, the nucleus of the cerebral cortex was tightly arranged and the number of nuclei observed under the microscope was increased significantly in each exercise group. Exercise promoted the expressions of SOD, BDNF and synaptophysin SYN1 in the cortex of rats, and the expression levels of SOD and BDNF in aged rats were up-regulated significantly (P< 0.01). The expression level of SYN1 in rats was up-regulated significantly (P<0.05) in the young and aged rats. The expression of CaMK IIα in the cortex of middle-aged and aged rats was up-regulated (P<0.01), while the expression level of CaMK IIα in young rats was down-regulated (P<0.01). Exercise could up-regulate the expression level of AMPKα1 in the cortex of young rats (P< 0.05), but not in middle-aged and old-age rats. Exercise could up-regulate the expression of SirT2 in the cortex of rats in all age groups (P<0.05). Exercise up-regulated the expression of phosphoinositide 3-kinase (IP3R)/ protein kinase B 1(AKT1) /mTOR in the cortex of rats, among which young IP3R was significantly up-regulated (P<0.01) in the young group, mTOR was significantly up-regulated in young and middle-aged group (P<0.01), and mTOR was also significantly up-regulated in the aged group (P<0.05).@*CONCLUSION@#Endurance exercise up-regulates BDNF expression, regulates CaMKIIα signaling, activates AMPK signaling pathway and IP3R / AKT1 / mTOR signaling pathway, and improves synaptic plasticity in the cortex.


Subject(s)
Animals , Male , Rats , Age Factors , Cerebral Cortex , Physiology , Neuronal Plasticity , Physical Conditioning, Animal , Physical Endurance , Rats, Sprague-Dawley , Signal Transduction
3.
China Journal of Chinese Materia Medica ; (24): 2559-2565, 2019.
Article in Chinese | WPRIM | ID: wpr-773226

ABSTRACT

Small molecules with physiological or pharmacological activities need to interact with biological macromolecules in order to function in the body. As the protein with the highest proportion of plasma protein,serum albumin is the main protein binding to various endogenous or exogenous small molecules. Serum albumin interacts with small molecules in a reversible non-covalent manner and transports small molecules to target sites. Bovine serum albumin( BSA) is an ideal target protein for drug research because of its low cost and high homology with human serum albumin. The research on the interaction between drugs and BSA has become a hotspot in the fields of pharmacy,medicine,biology and chemistry. In this research,molecular docking method was used to study the interaction between three small ginsenosides with high pharmacological value( Rg_1,Rb_1,Ro) and bovine serum albumin( BSA),and the binding mode information of three ginsenosides interacting with BSA was obtained. The results of molecular docking showed that ginsenosides and amino acid residues in the active pocket of proteins could be combined by hydrophobic action,hydrogen bonding and electrostatic action. The interaction between small ginsenosides and bovine serum albumin is not the only form,and their interaction has many forms of force. The interaction between these molecules and various weak forces is the key factor for the stability of the complex. The results of this study can provide the structural information of computer simulation for the determination of the interaction patterns between active components and proteins of ginseng.


Subject(s)
Animals , Cattle , Binding Sites , Computer Simulation , Ginsenosides , Chemistry , Molecular Docking Simulation , Protein Binding , Serum Albumin, Bovine , Chemistry , Spectrometry, Fluorescence , Thermodynamics
4.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 372-380, 2019.
Article in English | WPRIM | ID: wpr-776874

ABSTRACT

Macrophages play an important role in inflammation, and excessive and chronic activation of macrophages leads to systemic inflammatory diseases, such as atherosclerosis and rheumatoid arthritis. In this paper, we explored the anti-inflammatory effect of broussonin E, a novel phenolic compound isolated from the barks ofBroussonetia kanzinoki, and its underlying molecular mechanisms. We discovered that Broussonin E could suppress the LPS-induced pro-inflammatory production in RAW264.7 cells, involving TNF-α, IL-1β, IL-6, COX-2 and iNOS. And broussonin E enhanced the expressions of anti-inflammatory mediators such as IL-10, CD206 and arginase-1 (Arg-1) in LPS-stimulated RAW264.7 cells. Further, we demonstrated that broussonin E inhibited the LPS-stimulated phosphorylation of ERK and p38 MAPK. Moreover, we found that broussonin E could activate janus kinase (JAK) 2, signal transducer and activator of transcription (STAT) 3. Downregulated pro-inflammatory cytokines and upregulated anti-inflammatory factors by broussonin E were abolished by using the inhibitor of JAK2-STAT3 pathway, WP1066. Taken together, our results showed that broussonin E could suppress inflammation by modulating macrophages activation statevia inhibiting the ERK and p38 MAPK and enhancing JAK2-STAT3 signaling pathway, and can be further developed as a promising drug for the treatment of inflammation-related diseases such as atherosclerosis.

5.
China Journal of Orthopaedics and Traumatology ; (12): 853-860, 2019.
Article in Chinese | WPRIM | ID: wpr-773820

ABSTRACT

OBJECTIVE@#To explore the effect of lentivirus-mediated BMP-2 overexpression plasmid transfection into bone marrow mesenchymal stem cells and silk fibroin scaffold on osteoblast transformation.@*METHODS@#The lentivirus BMP-2 overexpression vector was constructed, bone marrow mesenchymal stem cells were cultured, and the combined culture system of nuclear scaffolds was constructed. Alizarin red staining and alkaline phosphatase staining were used to detect the osteogenic transformation of bone marrow mesenchymal stem cells in vitro. Ten New Zealand white rabbits, weighing 3.2 to 4.5 kg(averaging 3.9 kg), aged (2.89±0.45) years old, were selected to construct the rabbit tibial defect model by drilling a conical tibial defect (5 mm in length, 2 mm in width and 3 mm in depth) with an oral drill. The repair of the tibial defect in the animal model was observed by HE staining. The experimental group was implanted with silk fibroin scaffold + BMP-2 overexpression vector bone marrow mesenchymal stem cell complex, while the negative control group was implanted with silk fibroin scaffold+non-transfected bone marrow mesenchymal stem cell complex.@*RESULTS@#Compared with the control group(silk fibroin scaffold+non-transfected bone marrow mesenchymal stem cells), the number of adherent cells on the surface of the scaffold in the experimental group(silk fibroin scaffold+transfected BMP-2 overexpression vector BMP-2 complex) increased significantly. Compared with the control group, the ECM secretion in the experimental group increased significantly. EDX analysis showed that the content of calcium ion was 0.22% in the control group and 0.86% in the experimental group, which showed that the ability of inducing calcium ion formation in the experimental group was stronger than that in the control group. Alizarin red staining of calcium nodules showed that there was no obvious change in the naked eye of the control group, and a small amount of calcium nodules could be seen under the microscope. In the experimental group, obvious red area staining was observed by naked eye, and a large number of calcium nodules were observed by microscopy. The results of alkaline phosphatase staining showed that there was no obvious change in the naked eye of the control group, and no obvious change in the microscopic observation. In the experimental group, purple area staining was observed by naked eyes, and ALP staining was strongly positive by microscopy. The combined culture system of silk fibroin scaffold and bone marrow mesenchymal stem cells can repair cartilage defects. The repair effect of BMP-2 bone marrow mesenchymal stem cells after transfection is obviously better than that of non-transfection group. HE staining showed that inflammatory cells decreased and scaffolds disappeared slightly in the control group. In the experimental group, inflammatory cells were significantly reduced, scaffolds disappeared and angiogenesis was observed.@*CONCLUSIONS@#Lentivirus-mediated BMP-2 overexpression plasmid can promote BMSC to differentiate into osteocytes and secrete more extracellular matrix containing Ca²⁺ to promote bone defect repair.


Subject(s)
Animals , Rabbits , Bone Marrow Cells , Bone Morphogenetic Protein 2 , Cells, Cultured , Fibroins , Lentivirus , Mesenchymal Stem Cells , Osteoblasts , Osteogenesis , Plasmids , Transfection
6.
International Journal of Traditional Chinese Medicine ; (6): 592-595, 2018.
Article in Chinese | WPRIM | ID: wpr-693653

ABSTRACT

Objective In order to observe the clinical curative effect of magnesium sulfate combined with Xingshen-Kaiqiao acupuncture for the diffuse axonal injury (DAI). Methods A total of 60 patients with DAI were divided into the Xingshen-Kaiqiao group and control group according to the random number table method (30 in each). The control group was given traditional Chinese medicine wash-out on the basis of conventional therapy, saline 100 ml+25% magnesium sulfate 8 ml and slow intravenous infusion, then 5% glucose 500 ml+25% magnesium sulfate 8 ml and slow intravenous drip; during the duration of the drug, the concentration of blood magnesium was strictly monitored and the dosage was adjusted as appropriate. And the Xingshen-Kaiqiao group was given acupuncture of Xingshen-Kaiqiao on the basic treatment of the control group. All treatments lasted 4 weeks. After treatment, the expression levels of IL-1β and TNF-α, GSC score, recovery of consciousness and GOS were evaluated and compared before and after treatment. Results After treatment, the IL-1β (62.38 ± 16.92 pg/ml vs. 88.37 ± 14.71 pg/ml, t=8.554), the TNF-α (3.59 ± 1.05 pg/ml vs. 4.06 ± 1.62 pg/ml, t=5.081) of Xingshen-Kaiqiao group were significantly lower than those of the control group (P<0.05). The GSC scores after 1st week (8.26 ± 0.76 vs. 6.83 ± 0.82, t=-5.036), 2nd week (10.61 ± 0.82 vs. 8.91 ± 0.35, t=-4.387), 3rd week (12.52 ± 1.07 vs. 10.95 ± 0.67, t=-5.212), 4th week (13.26 ± 1.08 vs. 11.58 ± 1.86, t=-5.031) of Xingshen-Kaiqiao group were significantly higher than those of the control group ( P<0.05). After treatment, the Xingshen-Kaiqiao group awakening rate was 70.0% (21/30) and the control group was 43.3% (13/30), and the difference in awakening rate was statistically significant (χ2=4.344, P=0.037). The awakening time (7.6 ± 3.1 d vs. 11.5 ± 4.9 d, t=6.586) of Xingshen-Kaiqiao group was significantly lower than the control group (P<0.05); the percentage of GOS was significantly higher than the control group (Z=-2.093, P=0.036). Conclusions Magnesium sulfate combined with acupuncture of Xingshen-Kaiqiao can reduce the awakening time, the death rate and showe more satisfactory clinical efficacy for the patients with DAI.

7.
Chinese Journal of Immunology ; (12): 450-453,459, 2018.
Article in Chinese | WPRIM | ID: wpr-702752

ABSTRACT

Regulatory T (Treg) cells is an indispensable subset of T lymphocyte with the ability of immunosuppression in the periphery.Thymus-derived Treg cells(CD4+CD25+FOXP3+Treg cells) play a fundamental role in maintaining immune homeostasis in vivo.Treg cells have been actively involved in the onset and development of major human diseases including malignant tumors, autoimmune diseases,infectious diseases,allergic diseases and graft versus host disease(GVHD).Exosomes are membranous vesicles of endosomal origin that released from multiple cells into the extracellular space.They are currently considered to be vehicles containing protein,RNA and MicroRNA which can been transferred to recipient cells to modulate their activity by cell-contact-independent mecha-nism.Exosomes have a great impact on the induction and proliferation of Treg cells,understanding the relation of them will lead to novel therapeutic approaches for cancer immunotherapy,treating autoimmunity,infection,allergy and organ transplantation.

8.
Journal of Xinxiang Medical College ; (12): 6-11, 2018.
Article in Chinese | WPRIM | ID: wpr-699459

ABSTRACT

Objective To investigate the effect of different concentrations of magnesium-calcium alloy extract on the expression of matrix metalloproteinase-9 (MMP9) and tissue inhibitor of metalloproteinase-3 (TIMP3) in human colonic epithelial NCM460 cells.Methods The different concentrations of extracts (the volume fraction was 10%,50% and 100% respectively) were made with magnesium-calcium alloy.The 5 × 106 L-1 NCM460 suspension was randomly divided into control group,experimental group 1,experimental group 2 and experimental group 3.The cells in the control group were cultured by 2 000 μL high glucose Dulbecco's modified Eagle's medium (containing 10% volume fraction of fetal bovine serum).The cells in the experimental group 1,2 and 3 were cultured by 2 000 μL magnesium-calcium alloy extract with volume fraction of 10%,50% and 100% respectively.The expressions of MMP9 and TIMP3 mRNA in NCM460 cells was detected by real-time fluorescence quantitative polymerase chain reaction,and the expression of MMP9 and TIMP3 protein in NCM460 cells was detected by Western blot at after one,three and five days of cultivation respectively.Results The expression of MMP9 mRNA and TIMP3 mRNA in NCM460 cells of the experimental group 1,2 and 3 was significantly lower than that in the control group after one day of cultivation (P < 0.05).After three and five days of cultivation,the expression of MMP9 mRNA in NCM460 cells of the experimental group 1 was significantly lower than that in the control group (P < 0.05),but the expression of MMP9 mRNA in the NCM460 cells of the experimental group 2 and 3 was significantly higher than that in the control group and the experimental group 1 (P < 0.05).The expression of MMP9 mRNA in NCM460 cells of the experimental group 3 was significantly higher than that in the experimental group 2 after five days of cultivation (P < 0.05).The expression of MMP9 mRNA in NCM460 cells of the experimental group 1,2 and 3 after three and five days of cultivation was significantly higher than that after one day of cultivation(P < 0.05).There was no significant difference in the expression of MMP9 mRNA in NCM460 cells of the experimental group 1 between three and five days of cultivation (P > 0.05).The expression of MMP9 mRNA in NCM460 cells of the experimental group 2 and 3 after five days of cultivation was significantly higher than that after three days of cultivation(P < 0.05).The expression of TIMP3 mRNA in NCM460 cells of the experimental group 2 and 3 was significantly higher than that in the experimental group 1 after one day of euhivation (P < 0.05).After three days of cultivation,the expression of TIMP3 mRNA in NCM460 cells of the experimental group 1,2 and 3 was significantly lower than that in the control group (P < 0.05);the expression of TIMP3 mRNA in NCM460 cells of the experimental group 2 was significantly lower than that in the experimental group 1 and 3 (P < 0.05).After five days of cultivation,the expression of TIMP3 mRNA in NCM460 cells of the experimental group 1,2 and 3 was significantly higher than that in the control group (P < 0.05).The expression of TIMP3 mRNA in NCM460 cells after three and five days of cultivation was significantly higher than that after one day of cultivation (P < 0.05),and the expression of TIMP3 after five days of cultivation was significantly higher than that after three days of cultivation in the experimental group 1 (P < 0.05).The expression of TIMP3 mRNA in NCM460 cells after three days of cultivation was significantly lower than that after one day of cultivation (P < 0.05),and the expression of TIMP3 after five days of cultivation was significantly higher than that after one and three days of cultivation in the experimental group 2 (P < 0.05).The expression of TIMP3 mRNA in NCM460 cells after five days of cultivation was significantly higher than that after one and three days of cultivation in the experimental group 3 (P < 0.05).After five days of cultivation,there was no significant difference in the expression of MMP9 protein in NCM460 cells between the experimental group 1 and control group (P > 0.05),the expression of MMP9 protein in NCM460 cells of the experimental group 2 and 3 was significantly higher than that in the control group and the experimental group 1 (P < 0.05),but there was no significant difference in the expression of MMP9 protein in NCM460 cells between the experimental group 2 and 3 (P > 0.05).After five days of cultivation,the expression of TIMP3 protein in NCM460 cells of the experimental group 1,2 and 3 was significantly higher than that in the control group (P <0.05);but there was no significant difference in the expression of TIMP3 protein in NCM460 cells among the experimental group 1,2and 3 (P > 0.05).Conclusions The high concentration of magnesium-calcium alloy extract has certain influence on the expression of MMP9 and TIMP3 gene in NCM460 cells,which may lead to the early inflammatory reaction,and the mechanism may be related to the calcium ion concentration in the extract.

9.
Journal of Sun Yat-sen University(Medical Sciences) ; (6): 269-273, 2018.
Article in Chinese | WPRIM | ID: wpr-712945

ABSTRACT

[Objective]To explore the diagnostic value of urine ErbB3 protein in clear cell renal cell carcinoma.[Methods]We collected 31 urine samples of clear cell renal cell carcinoma patients who received operations in the Urology Department of the First Affiliated Hospital of Sun Yat-Sen University from April 2,2016 to August 31,2016.Meanwhile we collected 50 urine samples of normal people as control.We tested the expression of urine ErbB3 protein in experimen-tal group and control group,and analyzed the differences between the two groups.Then we established the ROC curve of which diagnosing clear cell renal carcinoma by urine ErbB3 protein. Also,we analyzed the relation between ErbB3 pro-tein in urine and the patients'BMI,preoperative creatinine,tumor diameter and underlying diseases such as hyperten-sion and hyperglycemia.[Results]①The expression of urine ErbB3 protein in clear cell renal cell carcinoma group was significantly lower than normal group(P<0.001). ② When diagnosing clear cell renal carcinoma by ErbB3 protein,the AUC of ROC was 0.802(P<0.001). When setting the cutoff as 13.98 pg/mL,the max Youden index was 0.525,the sensitivity was 0.645 and the specificity was 0.880. The Kappa value of diagnostic test was 0.542(P<0.001). ③ There was no correlation between the ErbB3 content and patients'BMI,tumor diameter or preoperative creatinine by correlation analysis. Also,there was no correlation between the urine ErbB3 protein content and blood pressure or blood glucose.[Conclusion]The urine ErbB3 protein of clear cell renal cell carcinoma was significantly lower than normal people,and it is meaningful for applying urine ErbB3 protein to early diagnosis of clear cell renal cell carcinoma.

10.
China Journal of Chinese Materia Medica ; (24): 2084-2089, 2015.
Article in Chinese | WPRIM | ID: wpr-337979

ABSTRACT

The rhizome of Panax japonicus var. major have been used as the natural medicinal agent by Chinese traditional doctors for more than thousand years. Most of the therapeutic effects of P. japonicus var. major had been reported due to the presence of tetracyclic or pentacyclic triterpene saponins. In this study, Illumina pair-end RNA-sequencing and de novo splicing were done in order to understand the pathway of triterpenoid saponins in this species. The valid reads data of 15. 6 Gb were obtained. The 62 240 unigenes were finally obtained by de novo splicing. After annotation, we discovered 19 unigenes involved in ginsenoside backbone biosynthesis. Additionally, 69 unigenes and 18 unigenes were predicted to have potential function of cytochrome P450 and UDP-glycosyltransferase based on the annotation results, which may encode enzymes responsible for ginsenoside backbone modification. This study provides global expressed datas for P. japonicus var. major, which will contribute significantly to further genome-wide research and analysis for this species.


Subject(s)
Gene Expression Profiling , Panax , Genetics , Saponins , Sequence Analysis, RNA
11.
Acta Pharmaceutica Sinica ; (12): 1183-1188, 2013.
Article in Chinese | WPRIM | ID: wpr-259495

ABSTRACT

P2X7 is the most important subtype of the ATP receptors known so far. Recent investigations showed that the downstream signaling pathway of P2X7 is coupled with several key inflammatory molecules including IL-1beta and IL-18, this suggests P2X7 might have roles in the inflammatory diseases. Moreover, attenuation of P2X7 by selective antagonists in vitro and knockout mice in vivo reducing the inflammatory response indicated that P2X7 is a potential therapeutic target for inflammatory diseases. However, most previous studies on P2X7 were focused on nerve system diseases most, while its effects in inflammatory respiratory diseases, especially in asthma, chronic obstructive pulmonary disease (COPD) and lung cancer have been poorly investigated. In this paper, we reviewed the research progress on the structure, distribution, biological activities of P2X7 and its relationship with inflammatory respiratory diseases including asthma, COPD and lung cancer, along with the development of P2X7 antagonist as therapeutics.


Subject(s)
Animals , Humans , Mice , Asthma , Drug Therapy , Metabolism , Inflammation , Drug Therapy , Metabolism , Interleukin-18 , Metabolism , Interleukin-1beta , Metabolism , Lung Neoplasms , Drug Therapy , Metabolism , Polymorphism, Single Nucleotide , Pulmonary Disease, Chronic Obstructive , Drug Therapy , Metabolism , Purinergic P2X Receptor Antagonists , Therapeutic Uses , Receptors, Purinergic P2X7 , Chemistry , Genetics , Metabolism , Respiratory Tract Diseases , Drug Therapy , Metabolism
12.
Chinese Medical Journal ; (24): 3690-3693, 2013.
Article in English | WPRIM | ID: wpr-236188

ABSTRACT

<p><b>BACKGROUND</b>Von Hippel-Lindau disease (VHL), a heritable autosomal dominant disease characterized by neoplasia in multiple organ systems, has rarely been reported in Asia. We genetically investigated a unique Chinese family with VHL disease and performed an analysis of the VHL protein stability.</p><p><b>METHODS</b>Genomic deoxyribonucleic acid (DNA) extracted from peripheral blood was amplified by polymerase chain reaction (PCR) to three exons of the VHL gene in 9 members of the Chinese family with VHL disease. PCR products were directly sequenced. We estimated the effects of VHL gene mutation on the stability of pVHL, which is indicated by the free energy difference between the wild-type and the mutant protein (ΔΔG).</p><p><b>RESULTS</b>The Chinese family was classified as VHL type 1. Three family members, including two patients and a carrier, had a T to G heterozygotic missense mutation at nucleotide 515 of the VHL gene exon 1. This missense mutation resulted in the transition from leucine to arginine in amino acid 101 of the VHL protein. There was low stability of the VHL protein (the ΔΔG was 12.71 kcal/mol) caused by this missense mutation.</p><p><b>CONCLUSIONS</b>We first reported a family with this VHL gene mutation in Asia. This missense mutation is predicted to significantly reduce the stability of the VHL protein and contribute to the development of the renal cell carcinoma (RCC) phenotype displayed by this family. The genetic characterization and protein stability analysis of families with VHL disease are important for early diagnosis and prevention of the disease being passed on to their offspring.</p>


Subject(s)
Adult , Female , Humans , Male , Middle Aged , China , Mutation, Missense , Protein Stability , Von Hippel-Lindau Tumor Suppressor Protein , Chemistry , Genetics , von Hippel-Lindau Disease , Genetics
13.
China Journal of Chinese Materia Medica ; (24): 1747-1750, 2013.
Article in Chinese | WPRIM | ID: wpr-294030

ABSTRACT

Eight compounds were isolated from the leaves of Turpinia arguta by various chromatograhic techniques such as D101 macroporous resin, polyamide, Sephadex LH-20,and HPLC chromatography, and their structures were elucidated as rhoifolin (1), apigenin-7-O- [2"-O-alpha-L-rhamnopyranosyl-6"-O-alpha-L-rhamnopyranosyl] -beta-D-glucopyranoside (2), acacetin-7-O- [2"-O-alpha-L-rhamnopyranosyl-6"-O-beta-D-glucopyranosyl] -beta-D-glucopyranoside (3), acacetin-7-O- [2"-O-alpha-L-rhamnopyranosyl-6"-O-alpha-L-rhamnopyranosyl] -beta-D-glucopyranoside(neobudofficide, 4), luteolin-7-O-[2"-O-beta-D-glucopyranosyl] -beta-D-glucopyranoside (5), chrysoeiml-7-O-[2"-O-beta-D-glucopyranosyl] -beta-D-glucopyranoside (6), acacetin-7-O-alpha-L-rhamnopyranosyl-(1 --> 6) -O-beta-D-glucopyranoside (buddleoside, linarin, 7), and apigenin 6, 8-di-C-beta-D-glucopyranoside (8) on the basis of spectral data analysis. Compounds 3-8 were isolated from T. arguta for the first time. Compounds 2, 3 showed weak anti-inflammatory effect on LPS-stimulated RAW264.7 cell.


Subject(s)
Animals , Mice , Anti-Inflammatory Agents , Chemistry , Pharmacology , Cell Line , Drugs, Chinese Herbal , Chemistry , Pharmacology , Flavonoids , Chemistry , Pharmacology , Glycosides , Chemistry , Pharmacology , Magnoliopsida , Chemistry , Mass Spectrometry , Molecular Structure , Plant Leaves , Chemistry
14.
Braz. j. med. biol. res ; 45(8): 737-745, Aug. 2012. ilus, tab
Article in English | LILACS | ID: lil-643650

ABSTRACT

Reports remain insufficient on whether and how prostate-specific membrane antigen (PSMA) can influence in vivo osseous metastasis of prostate cancer (PCa). In the present study, the authors induced stable expression of PSMA in mouse PCa cell line RM-1. In vivo osseous metastasis was induced in 37 6-week-old female C57BL/6 mice weighing 22.45 ± 0.456 g. RM-1 cells were actively injected into the femoral bone cavity, leading to bilateral dissymmetry of bone density in the femoral bone. Tumor cells were also detected in bone tissue by pathological examination. The impact on bone density was demonstrated by the significant difference between animals injected with RM-PSMA cells (0.0738 ± 0.0185 g/cm²) and animals injected with RM-empty plasmid cells (0.0895 ± 0.0241 g/cm²). The lytic bone lesion of the RM-PSMA group (68.4%) was higher than that of the control group (27.8%). Immunohistochemistry showed that the expression of both vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) was distinctly higher in the RM-PSMA group than in the control group, while ELISA and Western blot assay indicated that VEGF and MMP-9 were higher in the RM-PSMA group compared to the control group (in vitro). Thus, the present study proposed and then confirmed for the first time that PSMA can promote in vivo osseous metastasis of PCa by increasing sclerotic destruction of PCa cells. Further analyses also suggested that PSMA functions positively on the invasive ability of RM-1 by increasing the expression of MMP-9 and VEGF by osseous metastases in vivo.


Subject(s)
Animals , Female , Male , Mice , Antigens, Surface/metabolism , Bone Neoplasms/secondary , Glutamate Carboxypeptidase II/metabolism , Prostatic Neoplasms/pathology , Antigens, Surface/pharmacology , Bone Density/drug effects , Bone Density/physiology , Bone Neoplasms/pathology , Cell Line, Tumor , Glutamate Carboxypeptidase II/pharmacology , Immunohistochemistry , Matrix Metalloproteinase 9/metabolism , Neoplasms, Experimental/metabolism , Neoplasms, Experimental/pathology , Prostatic Neoplasms/metabolism , Vascular Endothelial Growth Factor A/metabolism
15.
Chinese Medical Journal ; (24): 2784-2786, 2012.
Article in English | WPRIM | ID: wpr-244352

ABSTRACT

A patient referred to our hospital, diagnosed with left idiopathic chronic orchialgia, was evaluated with a thorough medical and psychiatric history, physical examination, scrotal ultrasound and magnetic resonance imaging. Conservative management failed. The patient had temporary pain relief after undergoing outpatient cord block three times. Microsurgical denervation of the left spermatic cord was operated in March, 2011. A pain questionnaire was used to determine efficacy before and after operation, and complete pain relief was noted at one week after operation. The follow up period was 12 months, at the end of which the pain score was still zero. No complications, including testicular atrophy and hydrocele, occurred. Microsurgical denervation of the spermatic cord can be a minimally invasive, safe and effective management option for treatment of idiopathic chronic orchialgia.


Subject(s)
Humans , Male , Middle Aged , Denervation , Methods , Spermatic Cord , General Surgery , Testicular Diseases , General Surgery
16.
Chinese Journal of Oncology ; (12): 61-64, 2012.
Article in Chinese | WPRIM | ID: wpr-335343

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the clinical characteristics and treatment of localized Castleman's disease (CD), and review the literatures to improve the diagnosis and management of this disease.</p><p><b>METHODS</b>The clinical symptoms, histopathology, CT, MRI findings and results of surgery in 20 patients with localized CD were evaluated retrospectively.</p><p><b>RESULTS</b>The average age of the patients was 37.7 years. The lesions were located in the retroperitoneal space (9 cases), mediastinum (7 cases), pelvic cavity (1 case), neck (1 case), upper arm (1 case), and axillary (1 case). All patients underwent surgical resection, including 9 cases for retroperitoneal resection (6 cases had open operation and 3 cases laparoscopic resection) and 7 cases for mediastinal resection (open operation in 5 cases and thoracoscopic resection in 2 cases). The Castleman's disease was confirmed by histopathology. There were hyaline vascular type of CD in 17 cases, plasma cell type of CD in 1 case, and mixed cellularity type of CD in 2 cases. The duration of follow-up ranged from 12 to 165 months for 16 cases. Among them 15 patients were alive without recurrence, and 1 case had recurrence in the primary site at 47 months after the operation.</p><p><b>CONCLUSIONS</b>Patients with Castleman's disease have no typical clinical symptoms and have normal laboratory results. The majority of patients are of hyaline vascular type of the disease. Imaging examination is helpful to diagnosis, and the final diagnosis depends on pathologic examination. Complete surgical resection of the tumor is the best treatment for localized Castleman's disease.</p>


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Castleman Disease , Diagnosis , Diagnostic Imaging , Pathology , General Surgery , Follow-Up Studies , Magnetic Resonance Imaging , Mediastinum , Recurrence , Retroperitoneal Space , Retrospective Studies , Tomography, X-Ray Computed , Treatment Outcome
17.
Chinese Medical Journal ; (24): 3681-3686, 2012.
Article in English | WPRIM | ID: wpr-256667

ABSTRACT

<p><b>BACKGROUND</b>Multiple recurrences are common in non-muscle invasive bladder cancer, but the risk of multiple recurrences has not been fully described. Identifying patients at high risk of multiple recurrences will help to select an optimal therapeutic strategy and to improve prognosis. This study was conducted to identify the risk factors for multiple recurrences of non-muscle invasive bladder cancer.</p><p><b>METHODS</b>We reviewed the clinical data of all patients with non-muscle invasive bladder cancer in our hospital between January 2003 and February 2010. Patients with at least one recurrence were included. Multivariate analysis was performed for theorized risk factors (age, gender, tumor stage, grade, size, location, number of lesions, adjuvant intra-vesical chemotherapy after transurethral resection, and recurrence-free survival after each resection) to clarify risk factors for multiple recurrences of non-muscle invasive bladder cancer.</p><p><b>RESULTS</b>Of the 278 patients with non-muscle invasive bladder cancer, 84 were with at least one recurrence and a total of 222 recurrences among them were followed up for 6 - 70 months (mean, 36.1 months). Recurrence-free survival after initial resection predicted the overall frequency of bladder cancer recurrence (risk ratio (RR) = 37.83, 95% confidence interval (CI) = 3.45 - 396.13, P = 0.001) and second recurrence (RR = 6.15, 95%CI = 1.28 - 29.57, P = 0.023). Similarly, recurrence-free survival after a second resection was the only significant risk factor for third recurrence (RR = 31.08, 95%CI = 2.53 - 381.47, P = 0.007). Moreover, recurrence-free survival after initial resection was the only significant factor to predict later progression to muscle invasive bladder cancer (RR = 8.62, 95%CI = 1.47 - 58.34, P = 0.001).</p><p><b>CONCLUSIONS</b>Recurrence-free survival after resection is an independent predictor of multiple recurrences of non-muscle invasive bladder cancer. The shorter the period between resection and recurrence is, the higher the risk of multiple recurrences.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Cystectomy , Methods , Neoplasm Recurrence, Local , Retrospective Studies , Risk Factors , Time Factors , Urethra , Urinary Bladder Neoplasms , Mortality , Pathology , General Surgery
18.
Journal of Medical Biomechanics ; (6): E021-E026, 2012.
Article in Chinese | WPRIM | ID: wpr-803976

ABSTRACT

Objective To develop a novel measurement system composed of micro-CT, mechanical loading device and digital volume correlation (DVC) technique, so as to measure the three-dimensional microstructural deformation field in bone tissue. Methods Uniaxial compression was applied on the specimen with the micromechanical loading device, and CT scans were also conducted while maintaining the same loads; then sequential CT images were matched and searched accordingly by DVC method to calculate the micro-displacement in the specimen along three directions before and after loading; repeated scanning of zero-displacement and rigid body translation were used to evaluate the accuracy and precision of the system. The three-dimensional distribution of displacement field in bovine cancellous bone was measured by the system. Results The result from repeated scanning of zero-displacement showed that the highest accuracy of measurement was performed in the loading direction and the precision was less than tenth of the CT resolution. The result of rigid body translation showed that the standard deviation was 0.001~0.002 μm. For cancellous bone specimen under the load of 600 N, the range of micro-displacement was 100.35~110.25 μm, with multilayer field distribution. Conclusions The accuracy and precision of this measurement system can meet the requirement of DVC method. It is proved that this system can be used for measuring the three-dimensional micro-deformation field in the cancellous bone and as a measurement platform for investigating the relationship between deformation distribution and structural response in bone tissue for the future research.

19.
National Journal of Andrology ; (12): 502-506, 2011.
Article in Chinese | WPRIM | ID: wpr-305856

ABSTRACT

<p><b>OBJECTIVE</b>To assess the role of transrectal ultrasonography (TRUS) in the etiological diagnosis of male obstructive azoospermia.</p><p><b>METHODS</b>We retrospectively analyzed the clinical data and TRUS findings of 695 patients with obstructive azoospermia from January 2007 to May 2009.</p><p><b>RESULTS</b>Concerning the etiology of obstructive azoospermia, the main TRUS findings included ejaculatory duct abnormality (29.2%), seminal vesicle abnormality (25.4%) and prostate midline cyst (18.5%). TRUS revealed 203 cases of ejaculatory duct dilation, 177 cases of seminal vesicle abnormality (including 108 with absence or agenesis and 51 with dilation of the seminal vesicle), and 128 cases of prostate midline cyst (including 75 with ejaculatory duct cyst and 39 with Müllerian cyst). Calcification of the verumontanum or ejaculatory duct was suspected to be the causes of obstructive azoospermia in 34 cases. However, no significant etiological abnormality was found in 153 cases. Obvious etiology was shown by TRUS in 78.0% of the patients.</p><p><b>CONCLUSION</b>TRUS can clearly display the structural abnormality of the ejaculatory duct and seminal vesicle, and provide important information on the etiology of male obstructive azoospermia.</p>


Subject(s)
Adult , Humans , Male , Middle Aged , Azoospermia , Diagnostic Imaging , Rectum , Diagnostic Imaging , Retrospective Studies , Ultrasonography
20.
Journal of Southern Medical University ; (12): 1268-1272, 2011.
Article in Chinese | WPRIM | ID: wpr-235146

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of platelet-derived growth factor-BB (PDGF-BB) in the degeneration of the hypertrophied ligamentum flavum (LF) in the lumbar spine.</p><p><b>METHODS</b>Surgical specimens of degenerative hypertrophied LF were obtained from 11 patients with lumbar spinal stenosis (LSS, mean age 57.8 years), with those from 10 age-matched patients with lumbar disc herniation (LDH, mean age 53.5 years) as the normal controls. The thickness of the LF was measured preoperatively by axial T1-weighted magnetic resonance imaging (MRI) at the facet joint level. Immunohistochemistry was employed to determine the expression of PDGF β and PDGF-BB in the LF. The mRNA and protein expressions of PDGF-BB in the LF were detected using real-time PCR and Western blotting, respectively.</p><p><b>RESULTS</b>The thickness of the LF was 5.30±1.12 mm in the degenerative group and 2.80±1.53 mm in the control group, showing a significant difference between the two groups (P<0.001). PDGF-β and PDGF-BB were positive in the fibroblasts in hypertrophied LF. The mRNA and protein expressions of PDGF-BB were significantly higher in the degenerative group than in the control group (P=0.013 and 0.023, respectively).</p><p><b>CONCLUSION</b>The high expression of PDGF-BB in the hypertrophied LF suggests its important role in the development of hypertrophy of LF in lumbar spinal canal stenosis.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Hypertrophy , Metabolism , Intervertebral Disc Degeneration , Metabolism , Pathology , Ligamentum Flavum , Metabolism , Pathology , Lumbar Vertebrae , Proto-Oncogene Proteins c-sis , Metabolism , Spinal Stenosis , Metabolism , Pathology
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