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1.
Neuroscience Bulletin ; (6): 263-274, 2022.
Article in English | WPRIM | ID: wpr-929087

ABSTRACT

Protein O-GlcNAcylation is a post-translational modification that links environmental stimuli with changes in intracellular signal pathways, and its disturbance has been found in neurodegenerative diseases and metabolic disorders. However, its role in the mesolimbic dopamine (DA) system, especially in the ventral tegmental area (VTA), needs to be elucidated. Here, we found that injection of Thiamet G, an O-GlcNAcase (OGA) inhibitor, in the VTA and nucleus accumbens (NAc) of mice, facilitated neuronal O-GlcNAcylation and decreased the operant response to sucrose as well as the latency to fall in rotarod test. Mice with DAergic neuron-specific knockout of O-GlcNAc transferase (OGT) displayed severe metabolic abnormalities and died within 4-8 weeks after birth. Furthermore, mice specifically overexpressing OGT in DAergic neurons in the VTA had learning defects in the operant response to sucrose, and impaired motor learning in the rotarod test. Instead, overexpression of OGT in GABAergic neurons in the VTA had no effect on these behaviors. These results suggest that protein O-GlcNAcylation of DAergic neurons in the VTA plays an important role in regulating the response to natural reward and motor learning in mice.


Subject(s)
Animals , Dopaminergic Neurons/physiology , GABAergic Neurons/physiology , Mice , Nucleus Accumbens/metabolism , Reward , Ventral Tegmental Area/metabolism
2.
International Eye Science ; (12): 2129-2133, 2020.
Article in Chinese | WPRIM | ID: wpr-829719

ABSTRACT

@#AIM: To study the application value of magnetic resonance(MR)high-definition readout segmentation of long variable echo-trains diffusion weighted imaging(RESOLVE-DWI)in the diagnosis of thyroid associated ophthalmopathy(TAO)and its predictive value for the activity of TAO activity.<p>METHODS: A total of 82 patients(155 eyes)with TAO admitted to the hospital from January 2017 to December 2019 were selected as TAO group. Another 50 patients(100 eyes)with Graves disease without TAO who were admitted during the same period were selected as control group. According to the clinical activity score(CAS)the patients with TAO were divided into active period and inactive period. All the participants were examined by magnetic resonance(MR). The difference of RESOLVE-DWI apparent diffusion coefficient(ADC)values between TAO Group and control group was compared. The receiver operating characteristic curve(ROC)was used to evaluate the diagnostic value of the ADC measurement of RESOLVE-DWI for TAO. Count the related factors that may affect the activity of TAO, and use Logistic regression analysis to clarify the risk factors. ROC curve was used to evaluated the predictive value of RESOLVE-DWI ADC for TAO activity.<p>RESULTS: The ADC value of the TAO group was greater than that of the control group(<i>P</i><0.001). ROC curve showed that the best cut-off point of RESOLVE-DWI ADC value in diagnosing TAO was 1.302×10<sup>-3</sup>mm<sup>2</sup>/s, and the sensitivity, specificity and AUC were 87.10%, 81.94% and 0.895, respectively. The age, grading standard promulgated by the American Thyroid Society(NOSPECS)and ADC value of TAO in active period were higher than those in inactive period, and the differences were statistically significant(<i>P</i><0.05). Logistic regression analysis showed that age, NOSPECS grade and ADC value were the risk factors of TAO activity, and the differences were statistically significant(<i>P</i><0.05). ROC curve analysis showed that the best cut-off point of RESOLVE-DWI ADC value in predicting TAO activity was 1.522×10<sup>-3</sup>mm<sup>2</sup>/s, and the sensitivity, specificity, and AUC were 82.58%, 76.77%, 0.801, respectively.<p>CONCLUSION: The best cut-off points of RESOLVE-DWI ADC value in diagnosing TAO and predicting activity are 1.302×10<sup>-3</sup>mm<sup>2</sup>/s and 1.522×10<sup>-3</sup>mm<sup>2</sup>/s, respectively, and use the best cut-off point for TAO diagnosis and activity sexual prediction has high clinical value.

3.
Article in Chinese | WPRIM | ID: wpr-873353

ABSTRACT

Objective:To investigate the anti-tumor efficacy, mechanism and safety of zeylenone on acute T lymphocytic leukemia. Method:In vitro, Molt-4 cells were treated with various concentrations of zeylenone (0.2, 0.4, 0.8, 1.6, 3.2 μmol·L-1) for 48 h, and the cell viability was measured with cell counting kit-8 (CCK-8) assay. nonobese diabetic-severce combined immunodeficient mice(NOD/SCID) mice were randomly divided into six groups: normal group, model group, vincristine group (1 mg·kg-1), low-dose zeylenone group (12.5 mg·kg-1), medium-dose zeylenone group (25 mg·kg-1), high-dose zeylenone group (50 mg·kg-1). With the exception of normal group, mice were pre-irradiated with 60Co and inoculated subcutaneously with Molt-4 cells to establish the Molt-4 xenograft model. Then NOD/SCID mice were sacrificed after 13 days of administration. The tumor inhibition rates, relative tumor growth rates and organ indexes were calculated. Hematoxylin and eosin (HE) staining was used to observe the pathological changes of liver and spleen tissues in mice. The expressions of phosphorylation signal transducer and activator of transcription (p-STAT3), B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax) and cysteine aspartate-specific protease-3 (Caspase-3) were detected in tumor tissues by Western blot. Result:In vitro, zeylenone had an obvious inhibitory effect on Molt-4 cells. IC50 values of zeylenone was 1.49 μmol·L-1. In vivo, compared with the model group, medium and high-dose zeylenone groups had significant tumor inhibition effects, with the inhibition rates of 50.24% and 60.75%, respectively (P<0.01). Additionally, liver and spleen injuries were slight in the above mentioned two groups compared with the vincristine group, indicating that zeylenone was safe. Western blot analysis showed that medium and high-dose zeylenone groups showed significant declines in proteins p-STAT3, Caspase-3 and Bcl-2, and marked increases in pro-apoptotic protein Bax compared with the model group (P<0.05, P<0.01). Conclusion:zeylenone could obviously inhibit the proliferation and induce the apoptosis of Molt-4 cells; and its mechanism may be related to the down-regulation of p-STAT3, Caspase-3, Bcl-2 and the up-regulation of Bax expressions. In addition, zeylenone had less damage to liver and spleen, and was safer than vincristine.

4.
Article in Chinese | WPRIM | ID: wpr-873107

ABSTRACT

Objective::To identify the active anti-tumor constituents of Benzoinum according to observation of the anti-tumor effect of chemical constituents from Benzoinum in vitro. Method::The 95%ethanol extract of Benzoinum was systematically separated by silica gel column chromatography, medium pressure liquid preparation chromatography and preparation liquid chromatography, and their structures were identified by physicochemical property and spectral data. Anti-tumor activities of the compounds of Benzoinum were screened by in vitro cells including human hepatoma cells in vitro (HepG2), human lung cancer cells (A549), human cervical cancer cells (HeLa), human breast cancer cells (MCF-7) and human prostate cancer cells (PC-3). Result::Fifteen compounds were isolated from Benzoinum and identified as myricadiol(1), 3-keto-oleanonic acid(2), (4E)-1, 5-bis(4-hydroxyphenyl)-1-methoxy-2-(methoxy-methyl)-4-pentene(3a and 3b), (E)-p-coumaryl alcohol γ-Ο-methyl ether(4), sesamin(5), 5-(3″benzoyloxypropyl)-7-methoxy-2-(3′, 4′-methylenedioxy phenyl)-benzofuran(6), dibutyl phthalate(7), methyl 4-hydroxy-3-methoxybenzoate(8), p-hydroxybenzaldehyde(9), p-hydroxyacetophenone(10), acetovanillone(11), 3-oxo-olean-11, 13(18)-dien-28, 19β-olide(12), vanillin(13), benzoic acid(14), and siaresinolic acid(15). Compounds 1 to 11 were isolated from the resin of Styrax tonkinensis for the first time. A part of these compounds had good anti-tumor activities. Among them, compound 2, 12 showed a strongest activity. Conclusion::The chemical constituents of Benzoinum have good prospects for the development and application of anti-tumor drugs.

5.
Acta Physiologica Sinica ; (6): 824-832, 2019.
Article in Chinese | WPRIM | ID: wpr-781393

ABSTRACT

Drugs of abuse leads to adaptive changes in the brain stress system, and produces negative affective states including aversion and anxiety after drug use is terminated. Corticotrophin-releasing hormone (CRH) is the main transmitter in control of response to stressors and is neuronal enriched in the central amygdala (CeA), a sub-region of the extended amygdala playing an important role in integrating emotional information and modulating stress response. The effect of CRH neurons in CeA on the negative emotions on morphine naïve and withdrawal mice is unclear. Thus, we utilized CRH-Cre transgenic mice injected with AAV-mediated Designer Receptors Exclusively Activated By Designer Drugs (DREADDs) to chemogenetically manipulate CRH neurons in CeA. And methods of behavior analysis, including conditioned place aversion (CPA), elevated plus maze and locomotor activity tests, were used to investigate morphine withdrawal-induced negative emotions in mice. The results showed that, inhibiting CRH neurons of CeA decreased the formation of morphine withdrawal-induced CPA, as well as the anxiety level of CRH-Cre mice. Furthermore, specifically activating CRH neurons in CeA evoked CPA and anxiety of morphine naïve mice. Neither inhibiting nor activating CRH neurons had effects on their locomotor activity. These results suggest that CRH neurons in CeA are involved in the mediation of morphine withdrawal-induced negative emotion in mice, providing a theoretical basis for drug addiction and relapse mechanism.


Subject(s)
Adrenocorticotropic Hormone , Animals , Central Amygdaloid Nucleus , Corticotropin-Releasing Hormone , Metabolism , Emotions , Physiology , Mice , Morphine , Metabolism , Neurons , Metabolism
6.
Acta Pharmaceutica Sinica ; (12): 2083-2088, 2019.
Article in Chinese | WPRIM | ID: wpr-780284

ABSTRACT

The morphological identification, chemical component analysis, and DNA barcode determination were investigated on Genus Mentha (including Mentha haplocalyx Briq., Mentha piperita Linn., Mentha spicata Linn. and Mentha cultivated) in order to reveal the origin of Herba Mentha as a drug, and ensure the accuracy in clinic application. The morphological characters, chemical composition analysis by GC-MS/MS and DNA content measure by polymerase chain reaction (PCR) were reported in this study for inter- or intra- species divergence. Based on the morphology, axillary verticillasters was recognized as the typical character for Mentha haplocalyx Briq. Carvone was used as an index component for chemical composition analysis of Mentha spicata Linn. Gene clustering analysis divided 22 batches of samples into two molecular groups. Mentha haplocalyx Briq. is distinguishably different from Mentha spicata Linn. Mentha piperita Linn. and other cultivated plants were distributed between these two species. The results obtained by morphological identification, chemical composition analysis, and DNA barcode determination show good correlations, but each identification method has its limit. In view of the fact that hybridization of the plants in Genus Mentha is common, identification relying on only one method is not recommended.

7.
Article in Chinese | WPRIM | ID: wpr-693733

ABSTRACT

Objective Brucine and strychnine monomer reference substance as extremely toxic substance had potential threat during transportation and utilization. In this study we investigated the homogeneity, stability and assignment accuracy of the mixture reference solution of strychnine and brucine, so as to provide reference for the quality control of extremely toxic chemical reference substances for Chinese medicine. Methods Following the assay in Chinese Pharmacopoeia volume I (2015), we prepared the mixture reference solution of brucine and strychnine, and investigated the solvents and the concentration of mixutre reference solution. The stability test lasted for 12 months. F-test was used for heterogeneity assay. Three researchers were involved for collaboration. Results Methanol and chloroform solution were selected as the solvents for the stability test. Results showed the difference was not statistically significant among various mixture solutions. The results of value assignment were 0.14 mg/mL for strychnine (sR = 0.5%)and 0.10 mg/mL for brucine (sR = 1.0%). The stability of mixture solution were better under the conditions of methanol solution at 4 ℃ or -20 ℃. Conclusion The results provide a possible way to develop the mixture solution in place of the monomer reference, and the mixture reference solution is expected for the quality control in the slices of Semen Strychni and its compound preparations.

8.
Acta Physiologica Sinica ; (6): 463-473, 2018.
Article in Chinese | WPRIM | ID: wpr-687806

ABSTRACT

Drug addiction is a chronic psychiatric disorder characterized by compulsive drug taking, and involves neuronal plasticity changes in multiple brain regions. The prelimbic cortex (PrL) is a key region of the dorsomedial prefrontal cortex and contains majority of pyramidal neurons. The excitatory projections from PrL play a very important role in the drug seeking behaviors. PrL also contains a small amount of GABAergic interneurons, which regulate the information integration and transmission of the pyramidal neurons. However, the roles of the GABAergic interneurons in PrL in drug-induced behavior changes are not clear. In the PrL, parvalbumin (PV) and somatostatin (SST) interneurons are two major GABAergic interneurons, which have been reported to regulate the activity of glutamatergic input, and form inhibitory synaptic transmission to regulate the output of downstream signals. Here, we used PV-Cre and SST-Cre mice combined with chemical genetics to explore the role of PV and SST interneurons in PrL in morphine-induced behavior changes. Our data showed that specific inhibiting SST interneurons in PrL significantly increased the anxiety level and decreased morphine-induced locomotor activity and the conditioned place preference (CPP) score. Instead, specific inhibiting PV interneurons in PrL had no effect on the anxiety level, morphine induced-locomotor activity and CPP. Our findings provide a new insight into the cellular and neuronal specific mechanism for drug addiction.

9.
Acta Pharmaceutica Sinica ; (12): 439-443, 2018.
Article in Chinese | WPRIM | ID: wpr-779894

ABSTRACT

The antioxidant activities of Vleriana jatamansi Jones were investigated and the relationship between the antioxidant effect and the chemical structure was explored. The free radical scavenging test, 2,2-diphenyl-1-picrylhydrazyl (DPPH•), was used to evaluate the antioxidant activities of the extracts of Vleriana jatamansi Jones with 0−100% menthol as extraction solvents. The polar and nonpolar HPLC conditions were conducted to isolate the main chemical compositions. The DPPH• tests were used in analysis of the free radical scavenging activities. Under polar HPLC separation conditions, 5 kinds of compounds were detected:chlorogenic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, hesperidin, and coffeic acid; under nonpolar HPLC separation conditions, acevaltrate, 1β-acevaltrate and valtrate were founded. Chlorogenic acid, 3,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid presented high DPPH• free radical scavenging activities. The results of antioxidant activity suggested that the coffee acyl from chlorogenic acid-like compounds had a high DPPH• free radical scavenging ability. Our investigation indicated that structure of the ortho hydroxyl phenol of chlorogenic acid-like compounds play a significant role in antioxidant activities. In addition, this work can also provide method and theory reference for improving the antioxidant activities of Vleriana jatamansi Jones.

10.
Acta Physiologica Sinica ; (6): 241-251, 2017.
Article in Chinese | WPRIM | ID: wpr-348278

ABSTRACT

Rac1 belongs to the family of Rho GTPases, and plays important roles in the brain function. It affects the cell migration and axon guidance via regulating the cytoskeleton and cellular morphology. However, the effect of its dynamic activation in regulating physiological function remains unclear. Recently, a photoactivatable analogue of Rac1 (PA-Rac1) has been developed, allowing the activation of Rac1 by the specific wavelength of light in living cells. Thus, we constructed recombinant adeno-associated virus (AAV) of PA-Rac1 and its light-insensitive mutant PA-Rac1-C450A under the control of the mouse glial fibrillary acidic protein (mGFAP) promoter to manipulate Rac1 activity in astrocytes by optical stimulation. Primary culture of hippocampal astrocytes was infected with the recombinant AAV-PA-Rac1 or AAV-PA-Rac1-C450A. Real-time fluorescence imaging showed that the cell membrane of the astrocyte expressing PA-Rac1 protruded near the light spot, while the astrocyte expressing PA-Rac1-C450A did not. We injected AAV-PA-Rac1 and AAV-PA-Rac1-C450A into dorsal hippocampus to investigate the role of the activation of Rac1 in regulating the associative learning. With optical stimulation, the PA-Rac1 group, rather than the PA-Rac1-C450A group, showed slower learning curve during the fear conditioning compared with the control group, indicating that activating astrocytic Rac1 blocks the formation of contextual memory. Our data suggest that the activation of Rac1 in dorsal hippocampal astrocyte plays an important role in the associative learning.


Subject(s)
Animals , Astrocytes , Physiology , Cell Membrane , Cell Movement , Conditioning, Classical , Cytoskeleton , Dependovirus , Fear , Hippocampus , Physiology , Memory , Mice , Mice, Inbred C57BL , Neuropeptides , Genetics , Physiology , Optogenetics , rac1 GTP-Binding Protein , Genetics , Physiology
11.
Chinese Journal of Pathophysiology ; (12): 1932-1937, 2017.
Article in Chinese | WPRIM | ID: wpr-667675

ABSTRACT

AIM:To observe the effects of sevoflurane preconditioning on brain injury in hypoxic mice and its possible mechanism. METHODS:Male C57BL/6J mice were randomly divided into control (C) group, hypoxia (H) group,2% sevoflurane preconditioning for 30 min + hypoxia(S1+H) group,2% sevoflurane preconditioning for 60 min+hypoxia (S2+H) group and 4% sevoflurane preconditioning for 30 min + hypoxia(S3+H) group. The hypoxia model was established by continuous inhalation of(6.5±0.1)% O2for 24 h. The sevoflurane preconditioning treatments,S1,S2 and S3,were conducted by inhalation of 2% sevoflurane for 30 min,2% sevoflurane for 60 min and 4% sevoflurane for 30 min,respectively,with the carrier of(21.0±0.5)% O2,followed by washout for 15 min and then hypoxia treatment. The histological changes of the hippocampal CA1 area were observed under light microscope and transmission electron micro-scope(TEM),and serum lactate dehydrogenase (LDH) activity was measured by colorimetric method. Furthermore, the protein levels of erythropoietin (EPO) and vascular endothelial growth factor(VEGF) in brain tissue homogenate were ex-amined by ELISA,and the content of malondialdehyde(MDA) and the activity of superoxide dismutase(SOD) and gluta-thione peroxidase(GPx) were measured by microplate reader. RESULTS:After hypoxia for 24 h,cell edema or pyknosis in the hippocampal CA1 area was observed in H group. Sevoflurane preconditioning reduced hypoxic injury, and the cell ultrastructure under TEM was significantly improved in S2+H group. Compared with C group,the serum LDH activity and the levels of EPO,VEGF and MDA in brain tissues were significantly increased in H group,while the activity of SOD and GPx decreased. After sevoflurane pretreatment,the serum LDH activity and the levels of EPO and VEGF in brain tissues were lower than those in H group,and the most significant difference was observed in S2+H group. Moreover, the MDA content and SOD activity decreased,and the GPx activity increased in the sevoflurane preconditioning groups. CONCLU-SION:Sevoflurane preconditioning attenuates brain injury in hypoxic mice by regulating antihypoxic protein synthesis and reducing oxidative stress.

12.
Acta Physiologica Sinica ; (6): 233-240, 2016.
Article in Chinese | WPRIM | ID: wpr-331661

ABSTRACT

Caloric diet, such as fat and sugar intake, has rewarding effects, and has been indicated to affect the responses to addictive substances in animal experiments. However, the possible association between sucrose reward and the motivation for addictive drugs remains to be elucidated. Thus, we carried out behavioral tests after sucrose self-administration training to determine the effects of sucrose experience on rats' motivation for cocaine, locomotor sensitivity to cocaine, basal locomotor activity, anxiety level, and associative learning ability. The sucrose-experienced (sucrose) group exhibited higher lever press, cocaine infusion and break point, as well as upshift of cocaine dose-response curve in cocaine self-administration test, as compared with the control (chow) group. Additionally, despite similar locomotor activity in open field test and comparable score in cocaine-induced conditioned place preference, the sucrose group showed higher cocaine-induced locomotor sensitivity as compared with the chow group. The anxiety level and the performance in vocal-cue induced fear memory were similar between these two groups in elevated plus maze and fear conditioning tests, respectively. Taken together, our work indicates that sucrose experience promotes the rats' motivation for cocaine.


Subject(s)
Animals , Cocaine , Conditioning, Classical , Conditioning, Operant , Memory , Motivation , Rats , Reward , Self Administration , Sucrose
13.
Article in English | WPRIM | ID: wpr-258846

ABSTRACT

<p><b>OBJECTIVE</b>To explore the relationship between different components of fine particulate matter (PM2.5) emitted from coal combustion and their cytotoxic effect in the vascular endothelial cells.</p><p><b>METHODS</b>Coal-fired PM(2.5) was sampled using a fixed-source dilution channel and flow sampler. The sample components were analyzed by ion chromatography and inductively coupled plasma atomic emission spectroscopy (ICP-AES). The PM(2.5) suspension was extracted using an ultrasonic water-bath method and then human umbilical vein endothelial cells (EA.hy926) were treated with various concentrations of the PM(2.5) suspension. Cell proliferation, oxidative DNA damage, and global DNA methylation levels were used to measure the cellular toxicity of PM(2.5) emitted from coal combustion.</p><p><b>RESULTS</b>Compared to other types of coal-fired PM(2.5) preparations, the PM2.5 suspension from Yinchuan coal had the highest cytotoxicity. PM(2.5) suspension from Datong coal had the highest toxic effect while that from Yinchuan coal had the lowest. Exposure to coal-fired PM(2.5) from Jingxi coal resulted in lower 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels. At the same dose, PM(2.5) emitted from coal combustion could produce more severe DNA impairment compared to that produced by carbon black. Cell survival rate was negatively correlated with chloride and potassium ions content. The 5-methylcytosine (5-mC) level was positively correlated with Mn and negatively correlated with Zn levels. The 8 OHdG% level was positively correlated with both Mn and Fe.</p><p><b>CONCLUSION</b>PM(2.5) emitted from coal combustion can decrease cell viability, increase global DNA methylation, and cause oxidative DNA damage in EA.hy926 cells. Metal components may be important factors that influence cellular toxicity.</p>


Subject(s)
Cell Proliferation , Coal Ash , Toxicity , DNA Damage , DNA Methylation , Human Umbilical Vein Endothelial Cells , Toxicity Tests
14.
Acta Pharmaceutica Sinica ; (12): 439-2016.
Article in Chinese | WPRIM | ID: wpr-779189

ABSTRACT

Free amino acids play a great role in traditional Chinese medicine in injections of animal products, as they may take part in peptide synthesis and exhibit a bioactivity in vivo. However, most of the national standards for drugs and peer-reviewed papers only focus on the total amount of amino acids after peptide hydrolysis. We compare the advantage and disadvantage among high performance thin layer chromatography (HPTLC), pre-column derivatization ultra performance liquid chromatography (UPLC) and ion chromatography. As a result, the HPTLC and pre-column derivatization UPLC are suitable for quality analysis, while there is high matrix influence for ion chromatography analysis. The verified pre-column derivatization UPLC method is utilized in quantitative analysis. 24 kinds of amino acid were detected by this method, and 8 of them were reported for the first time from the injection. The system has high repeatability and accuracy with LOD on the level of pmol·mL-1.

15.
Chinese Journal of Endemiology ; (6): 311-314, 2012.
Article in Chinese | WPRIM | ID: wpr-642373

ABSTRACT

ObjectiveTo investigate current iodine nutritional status of different groups of people in Harbin city, and to provide the basis for development of salt iodization standard and scientific iodine supplementation.MethodsThree urban districts and three surrounding counties were chosen in Harbin,2011.In each chosen urban district and county,one district office (township) was selected,and one residents committee (village) was chosen in each of the district office(township),and 30 households were selected by systematic sampling.Iodized salt,water iodine and iodine intake per capita were investigated.In each of the residents committee (village),20 adults aged 18 - 45,30 pregnant women or lactating women,and 100 school children aged 8 - 10 were selected.Urine samples were collected and urinary iodine level were tested.Salt iodine was determined by direct titration,water and urinary iodine by arsenic cerium catalytic spectrophotometry.Iodine uptake and iodine nutritional status of different populations in Harbin urban and rural areas were compared.ResultsThe edible rate of qunlified iodized salt were 93.3%(84/90) and 96.3%(156/162) in Harbin urban and rural residents,respectively,which were all greater than 90%,and the highest value of salt iodine were 38.3,46.0 mg/kg,respectively,in urban and rural areas,which all did not exceed the upper limit(50 mg/kg) of qualified iodized salt,but there were some samples of salt iodine content below the national standard(20 mg/kg).Water iodine value in urban and rural areas,even the highest value(9.40,8.40 μg/L),was failed to meet the national standard 10 μg/L; salt eaten by rural people perperson a day(8.33 g) was significantly higher than that of the urban people(7.03 g,Z=- 2.750,P < 0.01); in addition to rural children aged 8 - 10,whose urinary iodine value(228.6 μg/L) was higher,the values in urban and rural adults ( 111.3,195.6 μg/L),pregnant women ( 193.0,172.9 μg/L),lactating women ( 128.4,173.7 μg/L)and urban children ( 186.8 μg/L ) were all in appropriate level.The urinary iodine medians ( 195.6,228.6 μg/L )of adults and children in rural were significantly higher than that of urban adults and children(111.3,186.8 μg/L,Z =- 2.294,- 5.434,P < 0.05 or < 0.01,respectively).Population composition of iodine deficiency in both urban and rural adults,lactating and pregnant women[46.7%(28/60),21.6%(13/60) ; 21.1%(19/90),21.3% (18/89) ; 27.8% ( 25/90 ),42.2% (38/90) ] were significantly higher than that of the population composition with iodine excess[4.6%(4/60),5.0%(3/60) ; 16.7%(15/90),16.9%(15/89) ; 4.4%(4/90),0.0%(0/90)],but proportion of iodine excessive population in rural children [26.3%(79/300)] was significantly higher than proportion of iodine deficiency[5.6%(17/300)].ConclusionsThe natural environment of Harbin city is still in the iodinedeficient state.In addition to children in rural areas,the iodine intake and iodine nutrition level is basically appropriate; the risk of disease caused by iodine deficiency in adults,lactating and pregnant women is higher than by iodine excess,but the situation of children in rural is on the opposite.Therefore,we should strengthen the monitoring of different populations,and supplement iodine scientifically based on their need.

16.
National Journal of Andrology ; (12): 445-449, 2010.
Article in Chinese | WPRIM | ID: wpr-295043

ABSTRACT

<p><b>OBJECTIVE</b>NYD-SP5 is a newly cloned gene highly expressed in human testes, which consists of 3 598 nucleotides including a 1 027-amino acid open reading frame. It is a human-mouse homologous gene. The domain analysis indicated that the NYD-SP5 protein is a transmembrane protein. This study aimed to design and establish recombinant plasmids of small hairpin interfering RNA (shRNA) against NYD-SP5, and to pave the way for the analysis of the function of NYD-SP5 in the testis using the transgenic mouse model.</p><p><b>METHODS</b>Four sequences of oligonucleotides with the small hairpin structure were designed based on the NYD-SP5 mRNA sequence. Recombinant plasmids were constructed by cloning these oligonucleotides into pGPU6/GFP/Neo vectors. Interfering plasmids against GAPDH were established as positive controls and those targeting non-specific genes used as negative controls. The positive constructs were verified by enzyme digestion and sequencing.</p><p><b>RESULTS</b>Plasmid screening and sequencing showed the sequences of the recombinant plasmids to be the same as the shRNA transcribed sequences, which indicated the successful establishment of the recombinant vectors.</p><p><b>CONCLUSION</b>The shRNA expression vector targeting NYD-SP5 could be established successfully.</p>


Subject(s)
Animals , Base Sequence , Genetic Vectors , Humans , Male , Mice , Mice, Transgenic , Molecular Sequence Data , Plasmids , Proteins , Genetics , RNA, Small Interfering , Genetics
17.
Journal of Experimental Hematology ; (6): 1027-1030, 2010.
Article in Chinese | WPRIM | ID: wpr-237602

ABSTRACT

The study was aimed to evaluate the adverse effects of PAD (bortezomib + adriamycin + dexamethasone) and VAD (vincristine + adriamycin + dexamethasone) as chemotherapy regimens in multiple myeloma patients. 27 and 30 patients with multiple myeloma (MM) were enrolled in PAD and VAD groups respectively. MM patients accepted 3 - 5 cycles of VAD or PAD regimens. The type, degree and occurrence time of adverse reactions during the treatment were observed. The results showed that the rash was found in two patients only in PAD group, leucocytopenia, thrombocytopenia, peripheral neuropathy, infection, fatigue, nausea, constipation, and adverse effects of cortex hormone (hypertension, glycemia, hypokalemia, hyponatremia and acne) were found in the both two groups. The thrombosis was not observed in both two groups during treatment. Although statistical analysis indicated that only the incidence of thrombocytopenia was higher in PAD group than in VAD group with statistical difference but the incidence of leucocytopenia, peripheral neuropathy and infection in PAD group were higher than those in VAD group. Rash, constipation, peripheral neuropathy could be found in the first course of chemotherapy, while the others mostly emerged after 3 courses of treatment. The main reasons for the patients who's treatment was stopped include infection and intolerable peripheral neuropathy. Although peripheral neuropathy could be found in the two groups, but the chemotherapy was stopped only in 2 patients of PAD group after 3 cycles of treatment. It is concluded that compared with conventional VAD chemotherapy, PAD may improve therapeutic effect, but it may bring more severe toxicities to the patients with multiple myeloma.


Subject(s)
Adult , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Boronic Acids , Bortezomib , Dexamethasone , Therapeutic Uses , Doxorubicin , Therapeutic Uses , Female , Humans , Male , Middle Aged , Multiple Myeloma , Drug Therapy , Pyrazines , Vincristine , Therapeutic Uses
18.
Chinese Journal of Hematology ; (12): 404-408, 2009.
Article in Chinese | WPRIM | ID: wpr-314471

ABSTRACT

<p><b>OBJECTIVE</b>To explore the expansion method of high purity NK cells from human peripheral blood and explore the changes in biological functions of NK cells after ex vivo expansion.</p><p><b>METHODS</b>NK cells were isolated from peripheral blood mononuclear cells (PBMNCs) by using miniMACS (Magnetic cell-selection) and NK Cell Isolation Kit II, and cultured in SCEM (Stemline Hematopoietic Stem Cell Expansion Medium, Sigma) supplemented with 10% human AB serum and different combinations of interleukin (IL)-2 and/or IL-12, IL-15 for 15 days. Cultures were semi-exchanged with fresh media and cytokines every 3 days. Evaluation for cell expansion, phenotype, perforin and granzyme B mRNA expressions, and IFN-gamma secretion before and after the culture period.</p><p><b>RESULTS</b>CD3(-) CD56(+) cells concentration increased from (11.2 +/- 5.2)% to (94.2 +/- 3.5)%. In group IL-2 + IL-15 and IL-2 + IL-15 + IL-12 group, cells were expanded 50.5 +/- 4.3 and 52.3 +/- 6.7 - fold, respectively, being significantly higher than that in other three groups [(15.4 +/- 1.1 fold in IL-2 group, 19.9 +/- 3.9 fold in IL-2 + IL-12 group, 6.1 +/- 1.0 fold in control group)] (P<0.01), but no significant difference between each other (P>0.05). The purity of CD3(-) CD56(+) NK cells was over 94% in all groups except the control. The perforin and granzyme B mRNA expressions of expanded NK cells in four experimental groups were significantly higher than those of before expansion (P<0.01) and the expressions in IL-2 + IL-15 and in IL-2 + IL-12 + IL-15 group were significant higher than in other three groups (P<0.01) while no significant difference between each other (P>0.05). IFN-gamma levels in the supernatants of four experiment groups were significantly higher than that in control group (P<0.01) and its levels order was IL-2 + IL-15 + IL-12 group > IL-2 + IL-12 group > IL-2 + IL-15 group > IL-2 group (P<0.01).</p><p><b>CONCLUSION</b>High purity NK cells isolated by negative selection using miniMACS can be efficiently expanded with IL-2 + IL-15, and their biological functions were enhanced.</p>


Subject(s)
Cell Culture Techniques , Cell Proliferation , Cell Separation , Cells, Cultured , Granzymes , Metabolism , Humans , Interferon-gamma , Metabolism , Interleukin-12 , Pharmacology , Interleukin-15 , Pharmacology , Interleukin-2 , Pharmacology , Interleukins , Pharmacology , Killer Cells, Natural , Cell Biology , Allergy and Immunology , Metabolism , Perforin , Metabolism
19.
Article in Chinese | WPRIM | ID: wpr-333996

ABSTRACT

This study was purposed to explore the changes in biological functions of human peripheral blood derived NK Cells after ex vivo expansion with different combinations of interleukin IL2 and/or IL12, IL15. According to different combination of cytokines, cultured NK cells were divided into 4 groups: group IL2, group IL2 + IL12, group IL2 + IL15 and group IL2 + IL15 + IL12. The group in which NK cells were cultured without cytokines was used as control. The cytotoxicity of cultured NK cells to target K562 cells was determined by using cell counting kit-8; the level of IFN-gamma in supernatants of NK cell culture was detected by ELISA; the perforin and granzyme B mRNA expressions were assayed by competitive quantitative RT-PCR. The results showed that the cytotoxicity of expanded NK cells in groups cultured with cytokines at different E:T ratio was significantly higher than that in group without cytokines (p < 0.01), although the cytotoxicity of NK cells in IL2 + IL15 + IL12 group seem to be slightly higher than that in IL2 + IL15 group, but there was no statistic difference (p > 0.05). The IFN-gamma levels in the supernatants of NK cell culture in the presence of cytokines significantly increased, and the IFN-gamma levels in IL2 + IL15 + IL12 group and IL2 + IL12 group were significantly higher than that in others (p < 0.01). The expressions of perforin and granzyme B mRNA of expanded NK cells in groups cultured with cytokines was significantly higher than that in control group (p < 0.01), and was consistent with cytotoxicity of NK cells. It is concluded that there are differences in the functions of NK cells cultured with different cytokines. IL2 and IL15 have synergistic effect on strengthening cytotoxicity of NK cells and promoting cell expansion. However, the main function of IL12 promotes NK cells to secrete IFN-gamma, which plays a role in immunoregulation.


Subject(s)
Humans , Interferon-gamma , Bodily Secretions , Interleukin-12 , Pharmacology , Interleukin-15 , Pharmacology , Interleukin-2 , Pharmacology , K562 Cells , Killer Cells, Natural , Allergy and Immunology
20.
Article in Chinese | WPRIM | ID: wpr-253322

ABSTRACT

This study was aimed to evaluate the effects of recombinant human interleukin 11 (rhIL-11) and recombinant human granulocyte colony stimulating factor (rhG-CSF) in mobilization for autologous peripheral blood stem cell transplantation (APBSCT). 16 patients with non-Hodgkin's lymphoma or acute myeloblastic leukemia were given myelosuppressive chemotherapy, then were mobilized by using rhG-CSF 5 microg/(kg.d) for median 5.5 days and rhIL-11 50 microg/(kg.d) for median 4 days (experimental group) or rhG-CSF 5 microg/(kg.d) alone for median 5.5 days (control group). After mobilizing, the peripheral blood leucocyte and platelet counts, total mononuclear cells, CD34+ cells and CFU-GM counts in PBSC collection, and amount of apheresed platelet transfusion were assayed. The results showed that the peripheral blood leucocyte and platelet counts, total mononuclear cell, CD34+ cell and CFU-GM counts in PBSC collection were no significant difference between two groups (p>0.05). After APBSCT, the median time for neutrophil count>or=0.5x10(9)/L and the median time for platelet count>or=20x10(9)/L were 10.5 and 11.5 days in experimental group, while were 13 and 13 days in control group, respectively. The median amount of apheresed platelet transfusion was 3.5 unit in experimental group and 5 unit in control group. Data were significantly different between two groups (p<0.05). The adverse reactions of mobilization were mild fever, fatigue, symptoms like as common cold, poor appetite, dizziness, muscular soreness in experimental group, but only mild fever in control. These symptoms were well tolerated and overcome with drug withdrawal. It is concluded that the regimen of rhIL-11 in combination with rhG-CSF after myelosuppressive chemotherapy to mobilize PBSC is efficient and safe with rapid hematologic reconstitution and less platelet transfusions after APBSCT were used.


Subject(s)
Adolescent , Adult , Female , Granulocyte Colony-Stimulating Factor , Hematopoietic Stem Cell Mobilization , Methods , Humans , Interleukin-11 , Leukemia, Myeloid, Acute , Therapeutics , Lymphoma, Non-Hodgkin , Therapeutics , Male , Middle Aged , Peripheral Blood Stem Cell Transplantation , Methods , Recombinant Proteins , Transplantation, Autologous , Young Adult
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