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1.
China Journal of Chinese Materia Medica ; (24): 1760-1769, 2023.
Article in Chinese | WPRIM | ID: wpr-981393

ABSTRACT

The present study aimed to investigate the effect of diosgenin on mammalian target of rapamycin(mTOR), fatty acid synthase(FASN), hypoxia inducible factor-1α(HIF-1α), and vascular endothelial growth factor A(VEGFA) expression in liver tissues of rats with non-alcoholic fatty liver disease(NAFLD) and explore the mechanism of diosgenin on lipogenesis and inflammation in NAFLD. Forty male SD rats were divided into a normal group(n=8) fed on the normal diet and an experimental group(n=32) fed on the high-fat diet(HFD) for the induction of the NAFLD model. After modeling, the rats in the experimental group were randomly divided into an HFD group, a low-dose diosgenin group(150 mg·kg~(-1)·d~(-1)), a high-dose diosgenin group(300 mg·kg~(-1)·d~(-1)), and a simvastatin group(4 mg·kg~(-1)·d~(-1)), with eight rats in each group. The drugs were continuously given by gavage for eight weeks. The levels of triglyceride(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C), alanine transaminase(ALT), and aspartate transaminase(AST) in the serum were detected by the biochemical method. The content of TG and TC in the liver was detected by the enzyme method. Enzyme-linked immunosorbent assay(ELISA) was used to measure interleukin 1β(IL-1β) and tumor necrosis factor α(TNF-α) in the serum. Lipid accumulation in the liver was detected by oil red O staining. Pathological changes of liver tissues were detected by hematoxylin-eosin(HE) staining. The mRNA and protein expression levels of mTOR, FASN, HIF-1α, and VEGFA in the liver of rats were detected by real-time fluorescence-based quantitative polymerase chain reaction(PCR) and Western blot, respectively. Compared with the normal group, the HFD group showed elevated body weight and levels of TG, TC, LDL-C, ALT, AST, IL-1β, and TNF-α(P<0.01), increased lipid accumulation in the liver(P<0.01), obvious liver steatosis, up-regulated mRNA expression levels of mTOR, FASN, HIF-1α, and VEGFA(P<0.01), and increased protein expression levels of p-mTOR, FASN, HIF-1α, and VEGFA(P<0.01). Compared with the HFD group, the groups with drug treatment showed lowered body weight and levels of TG, TC, LDL-C, ALT, AST, IL-1β, and TNF-α(P<0.05, P<0.01), reduced lipid accumulation in the liver(P<0.01), improved liver steatosis, decreased mRNA expression levels of mTOR, FASN, HIF-1α, and VEGFA(P<0.05, P<0.01), and declining protein expression levels of p-mTOR, FASN, HIF-1α, and VEGFA(P<0.01). The therapeutic effect of the high-dose diosgenin group was superior to that of the low-dose diosgenin group and the simvastatin group. Diosgenin may reduce liver lipid synthesis and inflammation and potentiate by down-regulating the mTOR, FASN, HIF-1α, and VEGFA expression, playing an active role in preventing and treating NAFLD.


Subject(s)
Rats , Male , Animals , Non-alcoholic Fatty Liver Disease/metabolism , Vascular Endothelial Growth Factor A/metabolism , Tumor Necrosis Factor-alpha/metabolism , Cholesterol, LDL , Rats, Sprague-Dawley , Liver , Inflammation/metabolism , Diet, High-Fat/adverse effects , TOR Serine-Threonine Kinases/metabolism , RNA, Messenger/metabolism , Body Weight , Mammals
2.
China Journal of Chinese Materia Medica ; (24): 5304-5314, 2023.
Article in Chinese | WPRIM | ID: wpr-1008728

ABSTRACT

This study aims to observe the effects of diosgenin on the expression of mammalian target of rapamycin(mTOR), sterol regulatory element-binding protein-1c(SREBP-1c), heat shock protein 60(HSP60), medium-chain acyl-CoA dehydrogenase(MCAD), and short-chain acyl-CoA dehydrogenase(SCAD) in the liver tissue of the rat model of non-alcoholic fatty liver disease(NAFLD) and explore the mechanism of diosgenin in alleviating NAFLD. Forty male SD rats were randomized into five groups: a control group, a model group, low-(150 mg·kg~(-1)·d~(-1)) and high-dose(300 mg·kg~(-1)·d~(-1)) diosgenin groups, and a simvastatin(4 mg·kg~(-1)·d~(-1)) group. The rats in the control group were fed with a normal diet, while those in the other four groups were fed with a high-fat diet. After feeding for 8 weeks, the body weight of rats in the high-fat diet groups increased significantly. After that, the rats were administrated with the corresponding dose of diosgenin or simvastatin by gavage every day for 8 weeks. The levels of triglyceride(TG), total cholesterol(TC), alanine transaminase(ALT), and aspartate transaminase(AST) in the serum were determined by the biochemical method. The levels of TG and TC in the liver were measured by the enzyme method. Oil-red O staining was employed to detect the lipid accumulation, and hematoxylin-eosin(HE) staining to detect the pathological changes in the liver tissue. The mRNA and protein levels of mTOR, SREBP-1c, HSP60, MCAD, and SCAD in the liver tissue of rats were determined by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR) and Western blot, respectively. Compared with the control group, the model group showed increased body weight, food uptake, liver index, TG, TC, ALT, and AST levels in the serum, TG and TC levels in the liver, lipid deposition in the liver, obvious hepatic steatosis, up-regulated mRNA and protein expression levels of mTOR and SREBP-1c, and down-regulated mRNA and protein expression levels of HSP60, MCAD, and SCAD. Compared with the model group, the rats in each treatment group showed obviously decreased body weight, food uptake, liver index, TG, TC, ALT, and AST levels in the serum, TG and TC levels in the liver, lessened lipid deposition in the liver, ameliorated hepatic steatosis, down-regulated mRNA and protein le-vels of mTOR and SREBP-1c, and up-regulated mRNA and protein levels of HSP60, MCAD, and SCAD. The high-dose diosgenin outperformed the low-dose diosgenin and simvastatin. Diosgenin may prevent and treat NAFLD by inhibiting the expression of mTOR and SREBP-1c and promoting the expression of HSP60, MCAD, and SCAD to reduce lipid synthesis, improving mitochondrial function, and promoting fatty acid β oxidation in the liver.


Subject(s)
Rats , Male , Animals , Non-alcoholic Fatty Liver Disease/genetics , Sterol Regulatory Element Binding Protein 1/metabolism , Diet, High-Fat/adverse effects , Diosgenin/metabolism , Chaperonin 60/therapeutic use , Rats, Sprague-Dawley , Liver , Signal Transduction , TOR Serine-Threonine Kinases/metabolism , Triglycerides , RNA, Messenger/metabolism , Simvastatin/therapeutic use , Body Weight , Lipid Metabolism , Mammals/metabolism
3.
Chinese Journal of Radiation Oncology ; (6): 589-593, 2022.
Article in Chinese | WPRIM | ID: wpr-932711

ABSTRACT

Radiation-induced brain injury (RBI) is one of the complications after radiotherapy for head and neck malignant tumors, which seriously affects the quality of life of patients. The pathophysiological mechanism of RBI is not completely clear. Current studies suggest that it is involved in a variety of cells in the central nervous system (CNS), whereas astrocyte, as the largest number of glial cells in the CNS, plays an important role in maintaining the CNS homeostasis and responding to CNS injury. In this article, the role of astrocytes in RBI was reviewed.

4.
Chinese Medical Journal ; (24): 269-276, 2020.
Article in English | WPRIM | ID: wpr-781574

ABSTRACT

BACKGROUND@#China's two-child policy has led to a trend of aging in pregnancy which was associated with adverse outcomes. This study aimed to identify the clinically cutoff maternal age for adverse obstetric outcomes in China.@*METHODS@#This secondary analysis of a multicenter retrospective cohort study included data of childbearing women from 39 hospitals collected in urban China during 2011 to 2012. Logistic regression was used to assess the adjusted odds ratios (aOR) of adverse outcomes in different age groups in comparison to women aged 20 to 24 years. The adjustments included the location of the hospital, educational level, and residence status. Clinically cutoff age was defined as the age above which the aOR continuously become both statistically (P  2) significant.@*RESULTS@#Overall, 108,059 women were recruited. In primiparae, clinically cutoff maternal ages for gestational diabetes (aOR: 2.136, 95% confidence interval [CI]: 1.856-2.458, P < 0.001), placenta previa (aOR: 2.400, 95% CI: 1.863-3.090, P < 0.001), cesarean section (aOR: 2.511, 95% CI: 2.341-2.694, P < 0.001), hypertensive disorder (aOR: 2.122, 95% CI: 1.753-2.569, P < 0.001), post-partum hemorrhage (aOR: 2.129, 95% CI: 1.334-3.397, P < 0.001), and low birth weight (aOR: 2.174, 95% CI: 1.615-2.927, P < 0.001) were 27, 31, 33, 37, 41, and 41 years, respectively. In multiparae, clinically cutoff ages for gestational diabetes (aOR: 2.977, 95%CI: 1.808-4.904, P < 0.001), hypertensive disorder (aOR: 2.555, 95% CI: 1.836-3.554, P < 0.001), cesarean section (aOR: 2.224, 95% CI: 1.952-2.534, P < 0.001), post-partum hemorrhage (aOR: 2.140, 95% CI: 1.472-3.110, P < 0.001), placenta previa (aOR: 2.272, 95% CI: 1.375-3.756, P < 0.001), macrosomia (aOR: 2.215, 95% CI: 1.552-3.161, P < 0.001), and neonatal asphyxia (aOR: 2.132, 95% CI: 1.461-3.110, P < 0.001) were 29, 31, 33, 35, 35, 41, and 41 years, respectively.@*CONCLUSIONS@#Early cutoff ages for gestational diabetes and cesarean section highlight a reasonable childbearing age in urban China. The various optimized cutoff ages for different adverse pregnancy outcomes should be carefully considered in childbearing women.

5.
Chinese Journal of Cancer Biotherapy ; (6): 410-415, 2020.
Article in Chinese | WPRIM | ID: wpr-821175

ABSTRACT

@#[Abstract] Objective:To explore the clinical significance of multiple serumcytokines in early diagnosis and progression assessment of gastric adenocarcinoma. Methods: Peripheral blood samples of 85 healthy subjects (healthy control group) and 81 patients with pathologically confirmed gastric adenocarcinoma (gastric cancer group) were collected from November 2017 to February 2018 at Shanxi Cancer Hospital. Serum levels of 17 cytokines (including IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-15, IL-17A, TNF-α, TNF-β, GM-CSF, G-CSF, IFN-γ, IP-10, MCP-1 andVEGF-A) were measured byAimPlex multiplex assay technology.Their diagnostic values were analyzed by receiver operating characteristic (ROC) curve. Results: Serum levels of IL-10, IL-8, IL-6, IP-10, MCP-1, VEGF-Aand IL-12p70 were significantly higher in gastric cancer patients than those in healthy controls (all P<0.01). There were significantly increasedlevelsofIL-8,IL-6and VEGF-Ain advanced-stage gastriccancer(stageI/II)groupoverearly-stage gastric cancer (stage III/IV) group (all P<0.01).AUC (areas under the curve) of IL-8, IL-6, IL-10, IP-10, MCP-1, IL-12p70 and VEGF-Afor distinguishing early-stage gastric cancer patientsfromhealthy controls was0.98,0.92,0.89,0.84,0.76,0.74 and 0.58, respectively. The diagnostic sensitivity of IL-8, IL-6 and IL-10 was 97.4%, 89.5% and 97.4%, respectively, and the specificity was 87.1%, 85.9%and 77.6%, respectively.TheAUCof IL-8, IL-6 andVEGF-Afor distinguishing advanced-stage gastric cancer patients from early-stage gastric cancer patients was 0.82, 0.72 and 0.69, respectively. Thediagnosticsensitivity of IL-8, IL-6 and VEGF-A was 83.7%, 60.5% and 41.9%, respectively, and the specificity was71.1%,76.3%and 92.1%, respectively. Conclusion: ThecombineddetectionofserumIL-8,IL-6andIL-10 may be a potential approach for early screening of gastric adenocarcinoma, which canalsobeusedtoassessthe progression of gastric adenocarcinoma.

6.
Chinese Journal of Surgery ; (12): 110-113, 2020.
Article in Chinese | WPRIM | ID: wpr-787668

ABSTRACT

To examine the effect of VAE and open surgery on the postoperativelocal recurrence of benign phyllodes tumors of breast and to investigate the clinical efficacy of VAE in the treatment of benign phyllodes tumors. The clinical data of 128 patients with benign phyllodes tumors of breast admitted to the Guangdong Women and Children Hospital from January 2013 to January 2018 were retrospectively analyzed. All patients were female, aged (37.7±9.1) years (range: 16 to 56 years). Eighty patients underwent ultrasound-guided VAE (minimally invasive group) and 48 patients underwent open surgery (open group). The -test, χ(2) test or Fisher exact probability method were used to compare the clinical characteristics of the two groups of patients. Logistic regression was used to analyze the prognostic factors of postoperative local recurrence. The maximum diameter of tumor in the minimally invasive group was smaller than that in the open group ((20.6±7.4) mm . (42.0±2.0) mm, -7.173, 0.000). The follow-up time was (36.4±1.8) months (range: 12 to 71 months). There were 7 cases of local recurrences during the follow-up period. The local recurrence rates in the minimally invasive and open groups were 5.0% (4/80) and 6.3% (3/48). The results of multivariate analysis showed that the maximum tumor diameter of 25 mm was an independent prognosis factor for postoperativelocal recurrence (0.122, 95: 0.016 to 0.901, 0.039). While surgical procedure, age, menopausal status and history of fibroadenomas in the ipsilateral breast is not an independent prognostic factor for postoperative local recurrence. In the minimally invasive surgery group, the local recurrence rates were 2.9% (2/69) and 2/11 in patients with tumor maximum diameters<25 mm and ≥25 mm, respectively. Local recurrence of breast benign phyllodes tumors is closely related to the tumor size. For patients with tumor diameter25 mm, the postoperative local recurrence rate of VAE is low, which can be used in clinical practice. Intraoperative complete resection to achieve a negative surgical margin should be guaranteed to avoid local recurrence.

7.
Biomedical and Environmental Sciences ; (12): 126-135, 2018.
Article in English | WPRIM | ID: wpr-776072

ABSTRACT

OBJECTIVE@#To better understand the pathological causes of bone loss in a space environment, including microgravity, ionizing radiation, and ultradian rhythms.@*METHODS@#Sprague Dawley (SD) rats were randomly divided into a baseline group, a control group, a hindlimb suspension group, a radiation group, a ultradian rhythms group and a combined-three-factor group. After four weeks of hindlimb suspension followed by X-ray exposure and/or ultradian rhythms, biomechanical properties, bone mineral density, histological analysis, microstructure parameters, and bone turnover markers were detected to evaluate bone loss in hindlimbs of rats.@*RESULTS@#Simulated microgravity or combined-three factors treatment led to a significant decrease in the biomechanical properties of bones, reduction in bone mineral density, and deterioration of trabecular parameters. Ionizing radiation exposure also showed adverse impact while ultradian rhythms had no significant effect on these outcomes. Decrease in the concentration of the turnover markers bone alkaline phosphatase (bALP), osteocalcin (OCN), and tartrate-resistant acid phosphatase-5b (TRAP-5b) in serum was in line with the changes in trabecular parameters.@*CONCLUSION@#Simulated microgravity is the main contributor of bone loss. Radiation also results in deleterious effects but ultradian rhythms has no significant effect. Combined-three factors treatment do not exacerbate bone loss when compared to simulated microgravity treatment alone.


Subject(s)
Animals , Biomechanical Phenomena , Bone Density , Physiology , Bone Resorption , Metabolism , Femur , Metabolism , Hindlimb Suspension , Rats, Sprague-Dawley , Tibia , Metabolism , Ultradian Rhythm , Weightlessness Simulation , X-Rays
8.
Biomedical and Environmental Sciences ; (12): 484-493, 2016.
Article in English | WPRIM | ID: wpr-296578

ABSTRACT

<p><b>OBJECTIVE</b>To explore the role of p21 in ionizing radiation-induced changes in protein levels during the G2/M transition and long-term G2 arrest.</p><p><b>METHODS</b>Protein expression levels were assessed by western blot in the human uveal melanoma 92-1 cells after treatment with ionizing radiation. Depletion of p21 was carried out by employing the siRNA technique. Cell cycle distribution was determined by flow cytometry combined with histone H3 phosphorylation at Ser28, an M-phase marker. Senescence was assessed by senescence- associated-β-galactosidase (SA-β-gal) staining combined with Ki67 staining, a cell proliferation marker.</p><p><b>RESULTS</b>Accompanying increased p21, the protein levels of G2/M transition genes declined significantly in 92-1 cells irradiated with 5 Gy of X-rays. Furthermore, these irradiated cells were blocked at the G2 phase followed by cellular senescence. Depletion of p21 rescued radiation-induced G2 arrest as demonstrated by the upregulation of G2/M transition kinases, as well as the high expression of histone H3 phosphorylated at Ser28. Knockdown of p21 resulted in entry into mitosis of irradiated 92-1 cells. However, cells with serious DNA damage failed to undergo cytokinesis, leading to the accumulation of multinucleated cells.</p><p><b>CONCLUSION</b>Our results indicated that p21 was responsible for the downregulation of G2/M transition regulatory proteins and the bypass of mitosis induced by irradiation. Downregulation of p21 by siRNA resulted in G2-arrested cells entering into mitosis with serious DNA damage. This is the first report on elucidating the role of p21 in the bypass of mitosis.</p>


Subject(s)
Humans , Cell Cycle Checkpoints , Radiation Effects , Cell Line, Tumor , Cyclin-Dependent Kinase Inhibitor p21 , Genetics , Metabolism , DNA Damage , Down-Regulation , Fibroblasts , Metabolism , Radiation Effects , Gene Expression Regulation , Radiation Effects , Mitosis , Radiation Effects , RNA Interference , RNA, Small Interfering , Radiation, Ionizing , Up-Regulation
9.
Acta Pharmaceutica Sinica ; (12): 824-829, 2015.
Article in Chinese | WPRIM | ID: wpr-257061

ABSTRACT

This paper is to report the exploration of the activation of Rho/ROCK signal pathway in 5-HT-induced proliferation of rat pulmonary artery smooth muscle cells (PASMCs) and the inhibitory effect of m-Nis on this pathway. PASMCs were cultured with the explant technique. MTT assay was used to explore the proliferation of PASMCs after 5-HT treated for different time and the intervening effect of m-Nis. RT-PCR and Western blot were used respectively to explore the mRNA expression of RhoA, ROCK1 and the protein expression of p-MYPT1 in 5-HT-treated PASMCs and intervening effect of m-Nis. The results of MTT assay suggested that 5-HT (1 µmol · L(-1)) treatment for 12-72 h significantly induced the proliferation of rat PASMCs (P<0.05 or P < 0.01), which were inhibited by m-Nis (1 x 10(-5), 1 x 10(-6), l x 10(-7), 1 x10(-8) mol · L(-1)) in dose-dependent manners (P < 0.05 or P < 0.01). Similarly, the mRNA expression of RhoA, ROCK1 and the protein expression of p-MYPT1 were also inhibited by m-Nis in different degrees (P < 0.05 or P < 0.01). Thus, the results of this study suggested that Rho/ROCK pathway played an important role in 5-HT-induced proliferation of rat PASMCs, m-Nis inhibited 5-HT-induced proliferation obviously, which may be related to the blockage of Rho/ROCK signal pathway.


Subject(s)
Animals , Rats , Cell Proliferation , Myocytes, Smooth Muscle , Cell Biology , Nisoldipine , Pharmacology , Protein Phosphatase 1 , Metabolism , Pulmonary Artery , Cell Biology , Serotonin , Pharmacology , Signal Transduction , rho-Associated Kinases , Metabolism , rhoA GTP-Binding Protein , Metabolism
10.
Shanghai Journal of Preventive Medicine ; (12): 113-115,127, 2014.
Article in Chinese | WPRIM | ID: wpr-789269

ABSTRACT

Objective] To analyze the effect of enhanced immunization for school-age children in Hongkou District of Shanghai in immunology and epidemiology . [ Methods] One month after completing immunization, the blood antibody titers were measured by ELISA method , and compared with the those of non-enhanced immunization , for epidemiological survey were collected history of measles immunization and close contact with the measles . [ Results] It could not be thought that strengthening immunization im-proved antibody positivity rate , the protection rate , and geometric antibody average concentrations .There was not statistical difference in antibody levels between measles vaccination 2 times and 3 -4 times. [ Conclusion] In the regions where routine immunization rates reach a high level , strengthening immuni-zation done on large scale is a waste of vaccine resources , human resources and financial resources , and leak re-vaccination should be done as a cost-effective preventive measure .

11.
Journal of Southern Medical University ; (12): 1575-1578, 2011.
Article in Chinese | WPRIM | ID: wpr-333860

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the aberrance of histone H3 lysine 4 trimethylation (H3K4me3) in patients with IgA nephropathy (IgAN).</p><p><b>METHODS</b>In 15 patients with IgAN and 15 healthy volunteers, H3K4me3 variations in peripheral blood mononuclear cells (PBMCs) were analyzed using chromatin immunoprecipitation and microarray analysis (ChIP-chip). ChIP real-time PCR was used to validate the microarray results. Quantitative real-time PCR (qRT-PCR) was carried out to examine the correlations between the mRNA expression profiles and H3K4me3 levels.</p><p><b>RESULTS</b>We identified 83 genes that displayed significant H3K4me3 differences in IgAN patients compared with healthy subjects. Among them, 39 genes showed increased H3K4me3 and 44 genes had decreased H3K4me3 levels. The results of ChIP real-time PCR were well consistent with the microarray data. Quantitative RT-PCR revealed the correlations between the mRNA expressions and the methylation levels of H3K4me3.</p><p><b>CONCLUSION</b>IgAN patients have significant alterations in H3K4me3, and the genes with aberrant H3K4me3 may provide insights into the pathogenesis of IgAN.</p>


Subject(s)
Female , Humans , Male , Case-Control Studies , CpG Islands , Genetics , DNA Methylation , Glomerulonephritis, IGA , Genetics , Metabolism , Histones , Genetics , Metabolism , Leukocytes, Mononuclear , Metabolism , Lysine , Genetics , Metabolism
12.
Chinese Journal of Oncology ; (12): 444-447, 2010.
Article in Chinese | WPRIM | ID: wpr-260381

ABSTRACT

<p><b>OBJECTIVE</b>To study the clinicopathological and immunohistochemical features, histogenesis and biological behavior, clinical treatment and prognosis of solid pseudopapillary tumor of the pancreas (SPT).</p><p><b>METHODS</b>Routine HE and immunohistochemical (SP) stainings were used in the pathological examination of 18 cases of SPT. Their clinical data were retrospectively analyzed. All the 18 postoperative patients were followed-up for 3 months to 10 years with an average of 29.2 months.</p><p><b>RESULTS</b>There were 16 females and 2 males, age ranging from 9 to 65 years with mean age of 25.3 years. Abdominal pain and palpable mass were among the major complains. Tumors were encapsulated and mixed with solid and cystic tissues. Histological features were pseudopapillary structure with a fibrovascular core. Immunhistologically, most tumors were positive for alpha-AT, alpha-ACT and Vim, with a high percentage of 94.4%. The eighteen cases were followed-up from 3 to 120 months. Five cases received reoperation after recurrence, and 14 cases were alive. Maximum survival time was 121 months and the minimum survival time was 3 months, with a median survival time of 23.0 months. The 5-year survival rate was 72.2%. A Kaplan-Meier analysis revealed that patient's age, tumor size, pathologic features, metastasis were major prognostic factors for SPT.</p><p><b>CONCLUSION</b>SPT is a tumor of low-grade malignancy and may be derived from multipotent stem cells. SPT most frequently affects young female, and has distinct clinicopathologic manifestation with excellent prognosis after surgical treatment.</p>


Subject(s)
Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Young Adult , Carcinoma, Papillary , Diagnosis , Metabolism , Pathology , General Surgery , Follow-Up Studies , Neoplasm Recurrence, Local , General Surgery , Pancreatectomy , Methods , Pancreatic Neoplasms , Diagnosis , Metabolism , Pathology , General Surgery , Reoperation , Retrospective Studies , Survival Rate , Vimentin , Metabolism , alpha 1-Antichymotrypsin , Metabolism , alpha 1-Antitrypsin , Metabolism
13.
Journal of Zhejiang University. Medical sciences ; (6): 71-78, 2010.
Article in Chinese | WPRIM | ID: wpr-259239

ABSTRACT

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus vector containing bone morphogenic protein-7 (BMP-7) gene and to identify its biological activities in proximal tubule epithelial cells (PTECs).</p><p><b>METHODS</b>Forty-six fragments of BMP-7 gene were obtained by PCR method and then were ligated to the full-length gene. The full length sequence of BMP-7 was subcloned into pBluescript II(+) vectors, and confirmed by sequencing. Double digested with Not I and Hind III, BMP-7 gene was inserted into pShuttle-CMV. EcoR I pre-linearized pShuttle plasmid was transformed into competence bacterium BJ5183 to obtain the recombinant adenovirus-BMP-7 by efficient homologous recombination. Then the AdBMP-7 was obtained by packaging Pac I linearized in 293 cells. Adenoviral titer was determined by adenovirus fluorescent detection kit. The protein expression of BMP-7 in PTECs was respectively detected by ELISA and Western blot. RT-PCR method was used for analyzing the alpha-smooth muscle action (alpha-SMA) expression in PTECs, which was treated consecutively with TGF-beta and AdBMP-7.</p><p><b>RESULTS</b>The recombinant plasmid AdBMP-7 was successfully generated, which increased BMP-7 protein expression levels in PTECs, and down-regulated TGF-beta-induced alpha-SMA expression.</p><p><b>CONCLUSION</b>The bioactive recombinant adenovirus AdBMP-7 has been successfully constructed, which may be effective in inhibition of chronic renal fibrosis.</p>


Subject(s)
Animals , Rats , Adenoviridae , Genetics , Metabolism , Base Sequence , Bone Morphogenetic Protein 7 , Genetics , Cells, Cultured , Epithelial Cells , Metabolism , Genetic Vectors , Genetics , Kidney Tubules, Proximal , Cell Biology , Metabolism , Molecular Sequence Data , Recombinant Proteins , Genetics
14.
Journal of Experimental Hematology ; (6): 1192-1197, 2009.
Article in Chinese | WPRIM | ID: wpr-343321

ABSTRACT

The aim of this study was to investigate the ability of calcium ionophore (CI ) to induce the differentiation of CML cells into dendritic cells (DC), to analyze the P210 expression in DCs and to evaluate the stimulatory effect of CML-DC on production of cytotoxic activity against CML cells via activating the autologous T cells. The mononuclear cells were isolated from bone marrow of CML patients whose WBC counts were more than 30x10(9)/L when samples were collected, then the lymphocytes and monocytes were discarded by pouring out supernatant twice at different culture time point. Slightly adherent cells were cultured in RPMI 1640 containing 10% FCS, with or without CI (375 ng/ml) and GM-CSF (200 ng/ml) at 37 degrees C, 5% CO2, fully humidified atmosphere for 96 hours. The cell morphology was observed under the inverted microscope and electron microscope; the expression of CD antigens was analyzed with flow cytometry; the P210 expression was measured with Western blot. LDH assay was used to evaluate the effect of cultured CML cells (CML-DC) generating cytotoxic T lymphocyte (CTL) activity against CML cells. The results indicated that after treatment with calcium ionophore and GM-CSF for 96 hours, CML cells showed DC morphological characteristics under inverted microscope and electron microscope. The expression of CD83, CD86, CD40, CD80 and HLA-DR increased remarkably. P210 was expressed in the CML-DC, but the expression level was lower than that in CML cells without CI and GM-CSF treatment. LDH assay showed that the CTL activity against CML was found greater in autologous T cells activated by CML-DC than that by CML cells. It is concluded that the CML cells can be induced to quickly differentiate into DC when cultured with CI and GM-CSF. CML-DC expresses P210, but the expression level is lower than that in CML cells. CML-DC can stimulate autologous T cells to produce CTL against CML.


Subject(s)
Female , Humans , Male , Antigens, CD , Metabolism , Bone Marrow Cells , Cell Biology , Calcium , Pharmacology , Cell Differentiation , Dendritic Cells , Cell Biology , Granulocyte-Macrophage Colony-Stimulating Factor , Pharmacology , Ionophores , Pharmacology , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Pathology , Monocytes , Cell Biology , Tumor Cells, Cultured
15.
Acta Pharmaceutica Sinica ; (12): 259-266, 2008.
Article in English | WPRIM | ID: wpr-277865

ABSTRACT

Effect of strophanthidin (Str) on intracellular calcium concentration ([Ca2+]i) was investigated on isolated ventricular myocytes of guinea pig. Single ventricular myocytes were obtained by enzymatic dissociation technique. Fluorescent signal of [Ca2+]i was detected with confocal microscopy after incubation of cardiomycytes in Tyrode' s solution with Fluo3-AM. The result showed that Str increased [Ca2+]i in a concentration-dependent manner. The ventricular myocytes began to round-up into a contracture state once the peak level of [Ca2+]i was achieved in the presence of Str (10 micromol L(- 1)), but remained no change in the presence of Str (1 and 100 nmol L(-1)). Tetrodotoxin (TTX), nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str (1 and 100 nmol L(-1)) , but had no obvious effects on the action of Str (10 micromol L(-1)). The elevation of [Ca2+]i caused by Str at all of the detected concentrations was partially antagonized by rynodine (10 micromol L(-1)) or the removal of Ca2+ from Tyrode's solution. In Na+, K+ -free Tyrode' s solution, the response of cardiomycytes in [Ca2+]i elevation to Str (10 micromol L(-1)) was attenuated, while remained no change to Str (1 and 100 nmol L(-1)). TTX, nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str at all of the detected concentrations in Na+, K+ -free Tyrode's solution. The study suggests that the elevation of [Ca2+]i by Str at the low (nomomolar) concentrations is partially mediated by the extracellular calcium influx through Ca2+ channel or a "slip mode conductance" of TTX sensitive Na+ channel. While the effect of Str at high (micromolar) concentrations was mainly due to the inhibition of Na+, K+ -ATPase. Directly triggering the release of intracellular Ca2+ from sarcoplasmic reticulum (SR) by Str may be also involved in the mechanism of [Ca2+]i elevation.


Subject(s)
Animals , 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester , Pharmacology , Aequorin , Pharmacology , Calcium , Metabolism , Calcium Channel Blockers , Pharmacology , Calcium Channels , Metabolism , Fura-2 , Pharmacology , Guinea Pigs , Myocardium , Pathology , Nifedipine , Pharmacology , Ryanodine , Pharmacology , Sarcolemma , Metabolism , Pathology , Sarcoplasmic Reticulum , Metabolism , Sodium-Calcium Exchanger , Sodium-Potassium-Exchanging ATPase , Strophanthidin , Pharmacology , Tetrodotoxin , Pharmacology , Thapsigargin , Pharmacology
16.
Chinese Journal of Biotechnology ; (12): 206-210, 2007.
Article in Chinese | WPRIM | ID: wpr-325392

ABSTRACT

S-adenosylmethionine-dependent uroporphyrinogen III methyltransferase (SUMT) is a novel red fluorescence indicator. However, the production of SUMT in Escherichia coli is restricted by its relatively low solubility, and little is known about the red fluorescent materials that are associate with SUMT. Two individual SUMT mutations, L166A and L88R/L89G double mutant were produced by site-directed mutagenesis. Both mutants were overexpressed in E. coli and purified by Ni-NTA chromatography. The reddish mixtures isolated from the purified L88R/L89G double mutant were analyzed by UV-visible spectra scanning and mass analysis(MS). The L88R/L89G double mutant has enzymatic activity in vivo, whereas L166A mutant loses the activity. Trimethylpyrrocorphin is identified as the main constituent in the isolated pigments. The purified L88R/L89G mutant increases protein solubility, which is applied potentially as the fluorescent indicator denoting the solubility of protein fusion partner.


Subject(s)
Amino Acid Substitution , Electrophoresis, Polyacrylamide Gel , Escherichia coli , Genetics , Fluorescence , Mass Spectrometry , Methyltransferases , Chemistry , Genetics , Metabolism , Molecular Weight , Mutagenesis, Site-Directed , Methods , Mutation , Pigments, Biological , Chemistry , Metabolism , Plant Proteins , Chemistry , Genetics , Metabolism , Recombinant Proteins , Chemistry , Metabolism , Solubility , Spectrophotometry, Ultraviolet , Zea mays , Genetics
17.
National Journal of Andrology ; (12): 134-137, 2007.
Article in Chinese | WPRIM | ID: wpr-289033

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the semen quality of the Chinese army men.</p><p><b>METHODS</b>Ten-item sperm quality analyses were made by manual methods and the computer assisted sperm analysis system in 1054 young Chinese army men. The subjects were divided into 4 age groups (18-20 yrs., 21-25 yrs., 26-30 yrs and 31-35 yrs.), and the results of the analyses were compared.</p><p><b>RESULTS</b>Among the 1 054 young males investigated, the semen volume was (2.6 +/- 1.4) ml, sperm density (55.9 +/- 46.5) x 10(6)/ml, sperm grade a + b motility (47.1 +/- 19.0)%, sperm viability (70.6 +/- 22.1)%, morphologically normal sperm (84.7 +/- 10.2)%, and acrosomal integrity (86.1 +/- 7.2)%. As for the percentages of the quality indexes that met WHO standards, the sperm volume was 73.5%, liquefaction time 91.1%, pH 93.0%, grade a + b motility 45.5%, viability 86.7%, sperm density 80.4%, morphologically normal sperm 98.2%, and the sperm total number 78.0%. Those who accorded with all the WHO standards accounted for 40.2%.</p><p><b>CONCLUSION</b>The semen quality of the 18-35 year old army men was better than previously reported in the similar literature. And that of the 26-30 yrs. group was the best among all the age groups.</p>


Subject(s)
Adolescent , Adult , Humans , Male , Hydrogen-Ion Concentration , Military Personnel , Sampling Studies , Semen , Chemistry , Physiology , Sperm Count , Sperm Motility
18.
National Journal of Andrology ; (12): 690-692, 2007.
Article in Chinese | WPRIM | ID: wpr-232084

ABSTRACT

<p><b>OBJECTIVE</b>To study the influence of varicocele on sperm chromatin structure and sperm motility.</p><p><b>METHODS</b>Routine semen analysis and sperm chromatin structure assay (SCSA) were performed in a varicocele group (n=74) and a control group (n=89).</p><p><b>RESULTS</b>Sperm concentration (41.4 +/- 38.7] x 10(6)/ml) grade a+b sperm percentage ([31.7 +/- 16.9]% and sperm viability ([62.8 +/- 22.2]%) in the varicocele group were evidently lower than those ([80.9 +/- 63.1] x 10(6)/ml, [46.8 +/- 20.5]%, [77.2 +/- 17.5])% in the control group (P < 0.05) and so were VCL, VSL and VAP ([37.4 +/- 12.5 microm/s, [23.4 +/- 7.8] microm/s, [26.5 +/- 8.2] microm/s) in the varicocele group than those ([42.4 +/- 10.7] microm/s, [27.3 +/- 7.3] microm/s, [30.7 +/- 7.8] microm/s) in the control (P < 0.05). MAD was increased (P < 0.01), and the COMP alphat of SCSA (23.2 +/-16.2) was obviously higher in the former than in the latter (14.1 +/- 11.8) (P < 0.05).</p><p><b>CONCLUSION</b>Varicocele causes damage to sperm DNA and changes sperm motility, which may result in male infertility.</p>


Subject(s)
Adolescent , Adult , Humans , Male , Chromatin , Genetics , Metabolism , DNA Damage , Sperm Count , Sperm Motility , Genetics , Physiology , Spermatozoa , Cell Biology , Metabolism , Varicocele , Genetics
19.
National Journal of Andrology ; (12): 306-308, 2007.
Article in Chinese | WPRIM | ID: wpr-297737

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the effect of Long-term exposure to low intensity microwave radiation on male reproductivity.</p><p><b>METHODS</b>A total of 289 married male radar operators were included in the radar group and 148 married men unexposed to microwave radiation were enrolled as controls. Questionnaires were used and the intensity of microwave radiation in different working areas was detected.</p><p><b>RESULTS</b>The rate of sexual dysfunction was 43.6% in the radar group and 24.4% in the control group (P < 0.01). The natural pregnancy rate was 53.6% within 1 year of marriage and 46.4% after 1 year of marriage in the radar group, as compared with 81.1% and 18.9% in the control group (P < 0.01).</p><p><b>CONCLUSION</b>Long-term exposure to low intensity microwave radiation evidently increased the sexual dysfunction rate and decreased natural pregnancy rate in men.</p>


Subject(s)
Female , Humans , Male , Pregnancy , Dose-Response Relationship, Radiation , Erectile Dysfunction , Epidemiology , Fertility , Radiation Effects , Microwaves , Military Personnel , Occupational Exposure , Pregnancy Outcome , Radar , Radiation Dosage , Surveys and Questionnaires
20.
China Journal of Chinese Materia Medica ; (24): 1188-1191, 2006.
Article in Chinese | WPRIM | ID: wpr-356709

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of Rongshi granule on osteopontin(OPN) expression.</p><p><b>METHOD</b>The urlisthiasis rats were induced by ethylene glycol (EG) and ammonium chloride, the control group rats were non-treated, and the Rongshi granule groups (low-dose group, middle-dose group and high-dose group) were administered Rongshi granule in addition to EG and ammonium chloride in 21 days. Pooled 24 h urine samples from each group were collected weekly with the use of metabolic cages, the concentration of uric calcium and oxalic acid were respectively measured by EDTA and photoelectric colorimetric method. Eight animals from each group were killed at 0, 7, 14, and 21 days, kidneys were histologic examinaed and immunohistochemical staining.</p><p><b>RESULT</b>The expression of kidney osteopontin in model group was obviously higher than that of control group (P <0.01), and was up to the highest at 21 days with 1.4 times (0.281 3/0.201 8) of the control group. The expression of kidney osteopontin in all of the Rongshi granule groups were lower than those of model group (P < 0.05), with an obvious dose-dependent manner. The degree of the kidney calcium oxalate crystal of the rats in all the Rongshi granule groups was much lower than that of model group, and the uric calcium and oxalic acid were much lower than those of model group (P < 0.01).</p><p><b>CONCLUSION</b>The Rongshi granule could inhibit the expression of osteopontin in rat urolithiasis model.</p>


Subject(s)
Animals , Female , Male , Rats , Ammonium Chloride , Calcium , Urine , Calcium Oxalate , Metabolism , Dose-Response Relationship, Drug , Drug Combinations , Drugs, Chinese Herbal , Pharmacology , Ethylene Glycol , Kidney , Metabolism , Kidney Calculi , Metabolism , Osteopontin , Metabolism , Oxalic Acid , Urine , Plants, Medicinal , Chemistry , Random Allocation , Rats, Sprague-Dawley
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