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1.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 61-68, 2022.
Artigo em Chinês | WPRIM | ID: wpr-940693

RESUMO

ObjectiveTo explore the intervention effect of Erxian decoction on intestinal microflora after ovariectomy in rats by 16S rRNA gene sequencing. MethodThirty-two female healthy SD rats were randomly divided into a Sham operation (Sham) group, a model (OVX) group, an estrogen (E) group, and an Erxian decoction (EXD) group, with 8 rats in each group. The rats in the E group and the EXD group received 1.8×10-4 g·kg-1 estradiol valerate solution and 9 g·kg-1 Erxian decoction, respectively, and those in the Sham group and the OVX group received an equal volume of distilled water once a day for 16 weeks. After 16 weeks, the levels of serum estrogen and blood lipid were detected. The fecal DNA was extracted, followed by 16S rRNA gene sequencing and analysis. ResultCompared with the Sham group, the OVX group showed reduced serum estrogen level (P<0.01) and increased serum levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) (P<0.05). Compared with the OVX group, the E group and the EXD group showed increased serum estrogen level (P<0.01) and reduced TC and LDL-C (P<0.05). Alpha diversity showed that there was no significant change in intestinal microflora diversity after ovariectomy. Beta diversity showed that there were significant differences in the structure of intestinal microflora in the four groups. The intervention of Erxian decoction could improve the changes in intestinal microflora after ovariectomy. LEfSe was used to analyze the differential flora in the four groups. The results showed that the Sham group and the OVX group had 3 differential bacterial phyla and 18 differential bacterial genera, the OVX group and the E group had 1 differential bacterial phylum and 12 differential bacterial genera, and the OVX group and the EXD group had 3 differential bacterial phyla and 5 differential bacterial genera. Estrogen intervention could reverse the change trend of Ruminococcus 1, Anaerovibrio, and Turicibacter in the OVX group. Erxian decoction intervention could reverse the change trend of Bacteroidetes, Firmicutes, Prevotella 9, Ruminococcaceae UCG-014, Ruminococcus 1, and Fusicatenibacter in the OVX group. ConclusionThe structure and function of intestinal microflora in ovariectomized rats changed obviously, and Erxian decoction could ameliorate the change.

2.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 9-18, 2022.
Artigo em Chinês | WPRIM | ID: wpr-940512

RESUMO

ObjectiveTo explore the effect of Qishengwan on ileal flora during its treatment of Alzheimer's disease (AD) under the guidance of the theory of "interior-exterior relationship between heart and small intestine". MethodThe AD model was established by bilateral intraventricular injection of β-amyloid 1-42 (Aβ1-42). The rats were then randomly divided into the blank group, sham-operated group, model group, low-, medium-, and high-dose (5.6, 11.2,22.4 g·kg-1·d-1) Qishengwan groups, and donepezil (0.46 mg·kg-1·d-1) group. After medication for 28 successive days, the spatial memory ability of rats was observed in water maze test, and the levels of Aβ1-42, nuclear transcription factor-κB (NF-κB), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) in the hippocampus were analyzed by enzyme-linked immunosorbent assay (ELISA). Additionally, the contents of the ileum were collected and subjected to 16SrRNA-sequencing analysis for figuring out the changes in ileal flora. ResultCompared with the blank group and sham-operated group, the model group exhibited significantly reduced stay time in the target quadrant and number of target quadrant and platform crossings (P<0.05, P<0.01) and elevated Aβ1-42 content in the hippocampus (P<0.01) and central inflammatory factors NF-κB, TNF-α, and IL-6 (P<0.05, P<0.01). Compared with the model group, Qishengwan at each dose significantly alleviated the impaired spatial memory function (P<0.05, P<0.01), improved the deposition of Aβ1-42 in the hippocampus of rats (P<0.05, P<0.01), and reduced the expression of central nervous system inflammatory factors (P<0.05, P<0.01), thus exerting a good therapeutic effect on AD rats. The 16SrRNA-sequencing analysis results showed that the structure of the ileal flora in the model group was significantly separated from those in the blank group and sham-operated group. The abundance of Lachnospiraceae NK4A136 group was significantly increased (P<0.01), while that of Escherichia-Shigella was reduced (P<0.05, P<0.01). Qishengwan at each dose significantly changed the ileal flora structure and regulated the relative abundance of Lachnospiraceae NK4A136 group, Escherichia-Shigella, and Ruminococcaceae. ConclusionQishengwan has a positive therapeutic effect on AD. It can significantly enhance the memory and cognitive abilities in AD rats, which may be related to its regulation of the structure of rat ileal flora and the relative abundance of Lachnospiraceae NK4A136 group, Escherichia-Shigella, and Ruminococcaceae, the attenuation of the central neuroinflammatory response, and the reduction of central Aβ1-42 deposition.

3.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 53-61, 2020.
Artigo em Chinês | WPRIM | ID: wpr-873121

RESUMO

Objective::To investigate the evolution of cardiac function and blood pressure in ovariectomized rats and the effect and mechanism of Erxiantang. Method::Healthy 10-week-old female SPF SD rats were randomly divided into sham operation group, model group, estrogen group(estradiol valerate, 0.18 mg·kg-1) and Erxiantang group(7.5 g·kg-1). The rats were intragastrically administered 2 weeks after ovariectomy, once a day for 12 weeks.Sham operation groups and model groups were given equal volumes of purified water.At the 4th week, 8th week, and 12th week after administration, the cardiac function, blood pressure, and levels of estrogen (E2) in rat serum were measured by non-invasive ultrasound cardiogram (UCG), tail artery detection techniques and radioimmunoassay.The levels of endothelin-1 (ET-1) and angiotensin 2(Ang Ⅱ) in rat serum were detected by enzyme-linked immuno sorbent assay (ELISA). The cardiac morphology and apoptosis were detected by hematoxylin-eosin (HE) staining, electron microscopy and Terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL). Result::Compared with sham operation group, the ejection fraction (EF) decreased and the left ventricular end systolic volume (LVVols) increased in the model group at 4th week after administration(P<0.05). There was no significant difference in cardiac function between the groups at 8th week.The left ventricular end diastolic diameter (LVIDs), LVVols, left ventricular end diastolic diameter (LVIDd), and left ventricular end diastolic volume (LVVold) were significantly increased in the model group at 12th week (P<0.01). At the 4th weeks, 8th week and 12th week, the systolic blood pressure (SBP) of the model group increased (P<0.05) and showed an increasing trend, and the diastolic blood pressure (DBP) did not change significantly.At the 12th week, the levels of E2 in serum decreased (P<0.05), ET-1 and Ang Ⅱ increased of the model group (P<0.01). The cardiac myofibrils were irregular, some myofilament was broken, and mitochondrial palsy was disordered, broken or disappeared, and cardiac apoptosis increased (P<0.01). Compared with the model group, myocardial contraction and diastolic function were significantly improved in Erxian decoction group, and blood pressure was decreased.The levels of E2 in serum was increased (P<0.05). The levels of ET-1 was decreased (P<0.05), and AngⅡ in serum was significantly decreased (P<0.01). The mitochondrial morphological structure was improved and the cardiac apoptotic rate was significantly decreased (P<0.01). Conclusion::After the ovariectomy, the rats showed a series of pathological changes such as decreased heart function and increased blood pressure.Compared with the decrease of heart function, the changes of blood pressure appeared earlier.Erxiantang exerts its intervention on cardiac function and blood pressure in ovariectomized rats by regulating E2, blood active substances and cardiac apoptosis.

4.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 59-67, 2020.
Artigo em Chinês | WPRIM | ID: wpr-872650

RESUMO

Objective:To observe the changes of myocardial microvessel density, microvascular endothelial cell morphology and hemorheology in ovariectomized rats and explore the interventional effects of Erxian decoction. Method:Thirty-two healthy 10 week-old female SPF SD rats were randomly divided into sham operation group, model group, estrogen group (estradiol valerate, 0.18 mg·kg-1) and Erxian decoction group (9 g·kg-1). The rats were intragastrically administered 2 weeks after ovariectomy, once a day for 16 weeks. Sham operation groups and model groups were given equal volumes of purified water. After 16 weeks of administration, the cardiac function was measured by noninvasive ultrasound cardiogram (UCG), CD34 in the myocardial tissue was tested by immunofluorescence staining to measure the microvessel density, the morphological structure of microvessels of myocardial tissue were detected by transmission electron microscope, the levels of estrogen (E2) in rat plasma were detected by radioimmunoassay, the levels of endothelin-1 (ET-1), prostacyclin I2 (PGI2), thromboxane A2 (TXA2), endothelial growth factor (VEGF), and von Willebrand Factor (vWF) in rat plasma were detected by enzyme-linked immuno sorbent assay (ELISA), four items of coagulation was detected by blood coagulation analyzer, whole blood viscosity and plasma viscosity were detected by hemorheology. Result:Compared with sham operation group, the ejection fraction (EF) decreased (P<0.01), the left ventricular short axis shortening rate (FS) decreased (P<0.01), and the left ventricular end systolic volume (LVVols) increased (P<0.01), myocardial microvessel density significantly reduced (P<0.01), the endothelial cells were swollen and the cytoplasm was cavitation, E2 in rat plasma decreased (P<0.01), ET-1, VEGF, vWF increased (P<0.01), prostacyclin I2 /thromboxane A2 (PGI2/TXA2) decreased (P<0.01), plasma activated partial prothrombin time (APTT) decreased (P<0.01), fibrinogen (FIB) increased (P<0.01), whole blood viscosity, plasma viscosity, and cassone viscosity increased (P<0.01), whole blood high-cut, low-cut index, and red blood cell (RBC) aggregation index increased (P<0.05) in model group. Compared with model group, EF and FS increased (P<0.05), LVVols decreased (P<0.05), myocardial microvessel density significantly increased (P<0.01), the endothelial cell edema was improved, and transport vesicles were clearly visible, E2 in rat plasma increased (P<0.01), ET-1, VEGF, decreased (P<0.01), PGI2/TXA2 increased (P<0.01), APTT increased (P<0.01), whole blood viscosity, whole blood high shear relative index, RBC aggregation index decreased (P<0.05), Kasson viscosity and plasma viscosity decreased (P<0.01) in Erxian decoction group. Conclusion:Erxian decoction increases myocardial microvessel density, protects the structural integrity of microvascular endothelial cells, improves its endothelial secretion function and hemorheology in ovariectomized rats, and protects heart function.

5.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 387-393, 2019.
Artigo em Inglês | WPRIM | ID: wpr-776872

RESUMO

Replacement of the native promoter of theglobal regulator LaeA-like gene of Daldinia eschscholzii by a strong gpdA promoter led to the generation of two novel cyclopentenone metabolites, named dalestones A and B, whose structures were assigned by a combination of spectroscopic analysis, modified Mosher's reaction, and electronic circular dichroism (ECD). Dalestones A and B inhibit the gene expression of TNF-α and IL-6 in LPS-induced RAW264.7 macrophages.

6.
Chinese Journal of Immunology ; (12): 769-773, 2018.
Artigo em Chinês | WPRIM | ID: wpr-702815

RESUMO

Ethylene carbodiimide (ECDI) is a chemical coupling agent,more and more studies have focused on the immune tolerance induced by administration of ethylene carbodiimide (ECDI)-fixed syngeneic splenocytes (Ag-SPs) coupled to peptides or to whole myelin proteins,in order to establish a novel and effective tolerance therapy for autoimmune diseases.The mechanisms and applications of immune tolerance induced by Ag-SPs in several kinds of common autoimmune diseases were summarized,and were compared with their counterparts in the traditional treatment.

7.
Chinese Medical Journal ; (24): 1034-1037, 2018.
Artigo em Inglês | WPRIM | ID: wpr-686986

RESUMO

<p><b>Background</b>Several studies have investigated the association between smoking and anal abscess and anal fistula (AA/F) diseases. However, the relationship between cigarette smoking and AA/F remains unclear. This study sought to assess the role of smoking in anorectal male patients in a Chinese population.</p><p><b>Methods</b>In this retrospective study, a questionnaire, including smoking history, was completed over a 3-month period by male inpatients in the Proctology Department of China-Japan Friendship Hospital. "Cases" were patients who had AA/F, and "controls" were patients with other anorectal complaints. Mann-Whitney U-test and Chi-square test were carried out to examine differences in baseline characteristics between groups. Subsequently, multivariate logistic regression was used to explore any related factors.</p><p><b>Results</b>A total of 977 patients aged from 18 to 80 years were included, excluding those diagnosed with inflammatory bowel disease or diabetes mellitus. Out of this total, 805 patients (82.4%) completed the entire questionnaire. Among the 805 patients, 334 (41.5%) were cases and 471 (58.5%) were controls. Results showed significant differences between cases and controls (χ = 205.2, P < 0.001), with smoking found to be associated with the development of AA/F diseases (odds ratio: 12.331, 95% confidence interval: 8.364-18.179, P < 0.001).</p><p><b>Conclusions</b>This study suggested smoking to be a potential risk factor for the development of AA/F diseases in a Chinese population. Consequently, current smoking patients should be informed of this relationship, and further research should be conducted to explore and investigate this further.</p>


Assuntos
Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Doenças do Ânus , Epidemiologia , Modelos Logísticos , Razão de Chances , Fístula Retal , Epidemiologia , Estudos Retrospectivos , Fatores de Risco , Fumar , Inquéritos e Questionários
8.
Chinese Hospital Management ; (12): 61-62, 2017.
Artigo em Chinês | WPRIM | ID: wpr-666876

RESUMO

The service system of Jingyitong has been started,the registration and payment online service has been implemented as well,which improves the efficiency of hospitals,patients' experience,and the environment for medical treatment to a certain extent.However,this system has brought certain internal control issues,especially the risk of financial security management.tn response to these risks,appropriate control measures could be taken to strengthen the internal control management and risk prevention.

9.
Journal of Forensic Medicine ; (6): 337-340, 2015.
Artigo em Chinês | WPRIM | ID: wpr-984006

RESUMO

OBJECTIVE@#To investigate the relationship between the expression of secreted frizzled-related protein 5 (SFRP5) mRNA and the time interval after skeletal muscle injury in rats by real-time PCR.@*METHODS@#A total of ninety SD rats were randomly divided into the contusion groups at different times including 4h, 8h, 12h, 16h, 20h, 24h, 28h, 32h, 36h, 40h, 44h, 48h after contusion, incision groups at different times including 4h and 8h after incision and the control group. The samples were taken from the contused zone at different time points. The total RNA was isolated from the samples and reversely transcribed to analyze the expression levels of SFRP5 mRNA.@*RESULTS@#Compared to the control group, the expression of SFRP5 mRNA in contusion groups were down-regulated within 48 h after contusion and reached the lowest level at 20 h, and the expression of SFRP5 mRNA gradually increased from 20 h to 48 h after contusion. The expression of SFRP5 mRNA in the incised groups were significantly lower than that of the contusion groups at 4 h after injury. At the time of 8 h, the expression levels between the contusion and incision groups showed no statistically significant difference.@*CONCLUSION@#It is suggested that SFRP5 mRNA analysis may show regular expression and can be a marker for estimation of skeletal muscle injury age.


Assuntos
Animais , Ratos , Biomarcadores/metabolismo , Contusões/metabolismo , Proteínas de Membrana/metabolismo , Músculo Esquelético/metabolismo , RNA Mensageiro , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real
10.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 197-202, 2014.
Artigo em Chinês | WPRIM | ID: wpr-312845

RESUMO

<p><b>OBJECTIVE</b>IBS-D rat model was established to assess the effect of Shen warming Pi strengthening method (SWPSM) for intervening diarrhea-predominant irritable bowel syndrome (IBS-D) by observing rats' general state, stool properties, AWR ranking, and histopathological changes.</p><p><b>METHODS</b>Totally 72 rats were randomly divided into 6 groups, i.e. the normal group, the model group, the high, middle, low dose SWPSM groups, and the control group, 12 in each group. The IBS-D rat model was successfully established referring to AL-Chaer ED's modeling method. After modeling high, middle, and low dose SWPS Recipe boil-free granules were given by gastrogavage to rats in corresponding treatment groups. Sishen Pill boil-free granule was given by gastrogavage to those in the control group. Equal volume of normal saline was given by gastrogavage to rats in the model group. The medication lasted for 2 weeks. Rats' general state, stool properties, abdominal withdrawal reflex (AWR) ranking, and histopathological changes were observed.</p><p><b>RESULTS</b>After treatment, the general state of all rats got im- provement to various degrees. The improvement in the high and middle dose SWPS Recipe groups were superior to that in the low dose SWPS Recipe group and the control group (P < 0.05). There was no statistical difference in the growth rate between after and before treatment in each group (P > 0.05). Compared with the model group and the low dose SWPS Recipe group, the defecation amount within 4 h was less in the high and middle dose SWPS Recipe groups and the control group (P < 0.05). The Bristol ranking score, average ranking of loose stool, ratio of dry stool and wet stool were lower in the high and middle dose SWPS Recipe groups than in the control group and the low dose SWPS Recipe group (P < 0.05). The AWR ranking score was lower in the high and middle dose SWPS Recipe groups than in the control group when the volume of balloon dilation was 1.5 mL. There was no organic change of histological or morphological observation.</p><p><b>CONCLUSIONS</b>High sensitive IBS-D model was proved to be reliable. SWPSM could reduce the quantity of stools, lower Bristol ranking score, average ranking of loose stools as well as ratios of dry stool and wet stool, contributing to reducing the high sensitivity of rats' visceral organs to some extent.</p>


Assuntos
Animais , Masculino , Ratos , Diarreia , Tratamento Farmacológico , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas , Usos Terapêuticos , Síndrome do Intestino Irritável , Tratamento Farmacológico , Fitoterapia , Ratos Sprague-Dawley
11.
China Journal of Chinese Materia Medica ; (24): 628-631, 2008.
Artigo em Chinês | WPRIM | ID: wpr-284375

RESUMO

<p><b>OBJECTIVE</b>To study scutellaria thermosensitive gel in situ.</p><p><b>METHOD</b>The phase transition temperature of gel with various concentration polymer solutions was studied by using stirring method and the viscosity of gel was monitored under different temperatures. Eye irritation experiments were performed with rabbit. The duration of residence time in rabbit eyes of scutellaria thermosensitive gel was observed with 2% fluorescein.</p><p><b>RESULT</b>20% poloxmar407 and 10% poloxmar 188 were suitable to scutellaria thermosen-sitive gel in situ. The results suggested that the scutellaria thermosensitive gel in situ based on poloxmar were nonirritant and retention time on the surface of eye was (150 +/- 8) min.</p><p><b>CONCLUSION</b>The scutellaria thermosensitive gel in situ forms gel in eye and has longer release time than that of eye drops.</p>


Assuntos
Animais , Feminino , Masculino , Coelhos , Medicamentos de Ervas Chinesas , Química , Metabolismo , Toxicidade , Olho , Géis , Soluções Oftálmicas , Metabolismo , Poloxâmero , Química , Scutellaria , Química , Metabolismo , Toxicidade , Sensibilidade e Especificidade , Fatores de Tempo , Temperatura de Transição
12.
China Journal of Chinese Materia Medica ; (24): 668-671, 2007.
Artigo em Chinês | WPRIM | ID: wpr-283413

RESUMO

<p><b>OBJECTIVE</b>To find the patterns of the rDNA ITS sequence variation of Crocus sativus, Chrysanthemum chanetii, Nelumbo nucifera, Zea mays and Garthamus tinctorius and to establish the molecular biological method for the identification of C. sativus and the others.</p><p><b>METHOD</b>After the total DNA of Crocus sativus, C. vernus-w and C. vernus-p were extracted, the ITS sequence was amplified by PCR with universal primer of ITS and PCR product was sequenced after purification and cloning. The ITS sequences of Chrysanthemrnum chanetii, Nelumbo nucifera, Zea mays and Garthamus tinctorius were obtained from GenBank.</p><p><b>RESULT</b>The complete ITS sequence of Crocus sativus, C. vernus-w and C. vernus-p, including ITSI rDNA, 5.8S rDNA, ITS2 rDNA were measured. The GenBank accession No. was DQ094185, DQ224363 and DQ224364 respectively. The similarity of ITS sequence between C. sativus and the two garden species of C. vernus was above 91%; the identity was 99.84% between C. vernus-w and C. vernus-p. The range of diversity between C. sativus and other herbs was above 46% based on ITS1 and above 41% based on ITS2.</p><p><b>CONCLUSION</b>C. sativus can be distinguished from misused substitutes by the ITS sequence. The ITS sequence is an available molecular marker for identification of the C. sativus.</p>


Assuntos
Chrysanthemum , Genética , Crocus , Genética , DNA de Plantas , Química , Genética , DNA Ribossômico , Química , Genética , DNA Espaçador Ribossômico , Química , Genética , Variação Genética , Dados de Sequência Molecular , Nelumbo , Genética , Plantas Medicinais , Genética , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Zea mays , Genética
13.
China Biotechnology ; (12)2006.
Artigo em Chinês | WPRIM | ID: wpr-685187

RESUMO

Recombinant antibodies have become the major growth trends in biotech industry following their success on therapeutic application and good revenue. But the low level of mammalian expression and laggard fermentation process constrained the development of antibody industry in China. The global advances of antibody industry were reviewed, compared the respective advantage between dihydrofolate reductase and glutamine synthetase expression system, continuous perfusion and fed-batch processes were compared. Finally, based on the knowledge and experience of antibody expression and fermentation, the suitable strategy of antibody industrialization, e.g. the fermentation model and scale, should depend on the comprehensive consideration of entrepreneur for the productivity, manufacturing capacity and market revenue. It may be a wise choice to use glutamine synthetase expression system and continuous perfusion process for the need of Chinese antibody industrialization.

14.
Journal of Southern Medical University ; (12): 294-296, 2006.
Artigo em Chinês | WPRIM | ID: wpr-255329

RESUMO

<p><b>OBJECTIVE</b>To compare the characteristics of three- and two-color flow cytometry for immunologic surveillance of toll-like receptor 4 (TLR-4) after renal transplantation.</p><p><b>METHOD</b>The expressions of CD14, TLR4 and CD80 in peripheral blood monocytes were measured in 10 patients after renal transplantation by three- and two-color fluorescent staining flow cytometry, respectively. The percentages of TLR4 and CD80 in CD14-positive monocytes were calculated, and the characteristics of two methods of flow cytometry were compared.</p><p><b>RESULTS</b>The two detection methods did not significantly differ in their detection of TLR4 (P=0.198) and CD80 (P=0.872) expressions in CD14-positive monocytes, and the measurements of the two methods for the same sample showed positive correlation (r=1, P=0.000 for TLR4 and r=0.999, P=0.000 for CD80). Three-color flow cytometry was also capable of detecting monocytes positive for both TLR4 and CD80. Compared with two-color flow cytometry, three-color flow cytometry provided more information and required less blood sample and monoclonal antibody.</p><p><b>CONCLUSIONS</b>Three- and two-color flow cytometry have almost the same accuracy in immunologic surveillance of TLR4 after renal transplantation. The former method also provides information of the simultaneous expression of TLR4 and its down stream molecule CD80, which could be critical for imploring the mechanisms by which TLR4 takes part in alloimmunity through modulating CD80 expression, and requires less blood sample and monoclonal antibody.</p>


Assuntos
Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Anticorpos Monoclonais , Química , Alergia e Imunologia , Antígeno B7-1 , Sangue , Alergia e Imunologia , Citometria de Fluxo , Métodos , Fluoresceína-5-Isotiocianato , Química , Transplante de Rim , Alergia e Imunologia , Receptores de Lipopolissacarídeos , Sangue , Alergia e Imunologia , Monócitos , Alergia e Imunologia , Reprodutibilidade dos Testes , Receptor 4 Toll-Like , Sangue , Alergia e Imunologia
15.
Journal of Southern Medical University ; (12): 1609-1615, 2006.
Artigo em Chinês | WPRIM | ID: wpr-232825

RESUMO

<p><b>OBJECTIVE</b>To prepare the recombinant murine Toll-like receptor-2 N-terminal (mTLR-2N) fusion protein and obtain anti-mTLR-2N polyclonal antibody.</p><p><b>METHODS</b>The gene encoding 153 amino acids of mTLR-2N was amplified by PCR and cloned into pET32A vector with sequence verification. The recombinant fusion protein was expressed in E. coli and purified by Probond resin column. Rabbits were immunized with fusion protein to obtain the polyclonal anti-sera, and the antibodies were identified by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry.</p><p><b>RESULTS</b>The recombinant fusion protein was efficiently expressed and purified. The polyclonal antibodies could bind to the fusion protein expressed in different vectors as the antigens in ELISA, and also bind with RAW264.7 cells expressing mTLR-2 and CHO cells transfected with full-length mTLR-2 gene.</p><p><b>CONCLUSION</b>The recombinant mTLR-2N fusion protein is obtained and the anti-mTLR-2N polyclonal antibody can recognize natural mTLR-2 on the cell surface.</p>


Assuntos
Animais , Cricetinae , Camundongos , Coelhos , Anticorpos Monoclonais , Células CHO , Linhagem Celular , Clonagem Molecular , Cricetulus , Ensaio de Imunoadsorção Enzimática , Escherichia coli , Genética , Vetores Genéticos , Soros Imunes , Alergia e Imunologia , Imuno-Histoquímica , Proteínas Recombinantes de Fusão , Alergia e Imunologia , Receptor 2 Toll-Like , Genética , Alergia e Imunologia , Transfecção
16.
Chinese Journal of Biotechnology ; (12): 43-48, 2004.
Artigo em Chinês | WPRIM | ID: wpr-305230

RESUMO

To construct the combined site-directed random mutation library of recombinant human Lymphotoxin (rhLT) for in vitro molecular evolution study, and to study the structure and function relationship. The random point mutations at the sites of 46,106 and 130 were individually generated by overlap PCR amplification with the random nucleotide primers. The three point mutations were combined and cloned into pMD-18T vector to construct the combined mutation library. DNA sequencing was used to evaluate the diversity and randomness of the mutation sites. The combined mutation library was re-engineered, inserted in prokaryotic expression vector pBV220, transformed and expressed in Escherichia coli strain DH5alpha. The biological activity of some of the mutants was tested in 1929 mouse fibroblast cells. As much as 1.5 x 10(5) clones were obtained, which represents 4.5 times of the complete mutation libraries at 99% confidence. Sequencing 50 clones revealed no obvious bias in the nucleotide and amino acid mutations at the sites. Among the 30 expressed samples underwent for the bioassay, 70% (21 samples) were inactive, 23.3% (7 samples) had lower activity than rhLT, the remaining 6.7% (2 samples) had higher activity than rhLT. The combined site-directed random mutation library of rhLT has been constructed successfully. In combination with phase display, the library is ready for in vitro molecular evolution study.


Assuntos
Humanos , Sequência de Aminoácidos , Sequência de Bases , Escherichia coli , Genética , Evolução Molecular , Biblioteca Gênica , Linfotoxina-alfa , Genética , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Proteínas Recombinantes , Genética
17.
Acta Academiae Medicinae Sinicae ; (6): 457-461, 2002.
Artigo em Chinês | WPRIM | ID: wpr-350085

RESUMO

<p><b>OBJECTIVE</b>To investigate the types of lipid disorder and its relationship with macrovascular diseases and insulin resistance in Chinese diabetic patients.</p><p><b>METHODS</b>2,430 diabetic patients finished the diabetes complications assessment and were surveyed on their blood lipids, insulin level and macrovascular diseases. The insulin resistance (HOMA-IR) was calculated based on HOMA model.</p><p><b>RESULTS</b>Prevalence of lipids disorder was as high as 63.8% in this group, including 23.9% with both hypercholesterolemia and hypertriglyceridemia (HY-C-T), 16.1% with hypercholesterolemia only (HY-C), 15.0% with hypertriglyceridemia only (HY-T), 5.5% with lower HDL level (L-HDL), 3.3% with hypercholesterolemia, hypertriglyceridemia and lower HDL level together. Compared with the patients with normal lipid level (control group), no duration differences existed among these groups. There were more female patients in HY-C and HY-C-T groups. BMI and WHR were higher in all groups with lipid disorder except in HY-C group. Macrovascular diseases were more common in HY-C-T patients than in control group (33.6% vs 24.0%, P < 0.001). Average blood pressure and the prevalence of hypertension were significantly higher in HY-C-T, HY-C, HY-T groups than in control group (139/79, 138/76, 134/77 vs. 132/75 mmHg; 53.2%, 50.1%, 46.2% vs. 39.2%). Fasting insulin level was significantly higher in HY-C-T and HY-C-T-L-LDL groups than in controls. Insulin resistance was more severe in all patients with lipid disorder except in L-HDL group, particularly in the HY-C-T-L-HDL group.</p><p><b>CONCLUSIONS</b>(1) BMI, WHR increased significantly with the severity of lipid disorder. (2) Nearly two thirds of Chinese diabetic patients have lipid disorder, which is the strong risk factor of macrovascular diseases and aggravates insulin resistance. (3) There was severe insulin resistance in patients with hypertriglyceridemia or with hypertriglyceridemia plus other lipid disorder.</p>


Assuntos
Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , China , Epidemiologia , Complicações do Diabetes , Nefropatias Diabéticas , Retinopatia Diabética , Hiperlipidemias , Epidemiologia , Hipertensão , Epidemiologia , Resistência à Insulina , Prevalência
18.
Academic Journal of Second Military Medical University ; (12): 32-35, 2001.
Artigo em Chinês | WPRIM | ID: wpr-736796

RESUMO

Objective: To investigate the role of T cell in the antitumor immune responses induced by MIF gene-modified tumor vaccine. Methods: MIF gene was transferred into FBL3 erythroleukemia cel l by adenovirus carrier and a new type of tumor vaccine was prepared. The chang es of the number and the function of T cell in spleen and lymph node was observe d. Results: After the mice were immunized with MIF gene-m odified FBL3 vaccine, the number of lymphocyte in spleens and lymph nodes increa sed markedly and the specific CTL activities of splenocytes also increased great ly. FACS analysis showed that the CD3+, CD4+, CD8+ T cells and CD28 posi tive cells in draining lymph nodes of MIF-FBL3 group mice increased more marked ly than that of control groups. When the wild type FBL3 cells were injected into the mice immunized with MIF gene-modified FBL3 vaccine, the growth of tumors w ere obviously inhibited and the survival rate of the mice was increased. Conclusion: It is suggested that MIF gene-modified tumor vaccine can induce specific antitumor immune responses mediated by T cells and may be a candidate for gene therapy of tumor.

19.
Academic Journal of Second Military Medical University ; (12): 32-35, 2001.
Artigo em Chinês | WPRIM | ID: wpr-735328

RESUMO

Objective: To investigate the role of T cell in the antitumor immune responses induced by MIF gene-modified tumor vaccine. Methods: MIF gene was transferred into FBL3 erythroleukemia cel l by adenovirus carrier and a new type of tumor vaccine was prepared. The chang es of the number and the function of T cell in spleen and lymph node was observe d. Results: After the mice were immunized with MIF gene-m odified FBL3 vaccine, the number of lymphocyte in spleens and lymph nodes increa sed markedly and the specific CTL activities of splenocytes also increased great ly. FACS analysis showed that the CD3+, CD4+, CD8+ T cells and CD28 posi tive cells in draining lymph nodes of MIF-FBL3 group mice increased more marked ly than that of control groups. When the wild type FBL3 cells were injected into the mice immunized with MIF gene-modified FBL3 vaccine, the growth of tumors w ere obviously inhibited and the survival rate of the mice was increased. Conclusion: It is suggested that MIF gene-modified tumor vaccine can induce specific antitumor immune responses mediated by T cells and may be a candidate for gene therapy of tumor.

20.
Chinese Journal of Radiology ; (12)2001.
Artigo em Chinês | WPRIM | ID: wpr-679743

RESUMO

Objective To determine the methods and results of treating traumatic pseudoaneurysms in siphon segment of internal carotid artery(ICA)by interventional therapy.Methods Twelve cases of traumatic pseudoaneurysms in siphon segment of internal carotid artery were treated.The collateral circulation of Willis circle was observed after DSA.Different methods of treatment were applied according to the collateral circulation of Willis circle.Ten cases were treated by occlusion of ICA completely,1 case was embolized by guglielmi detachable coil(GDC)only.Results Nine of 12 treated by occlusion of ICA were cured.In the 3 cases who had poor collateral of Willis circle,one was cured by GDC embolization alone;one died 48 hours later after ICA occlusion though his consciousness and the activity of extremities were normal during the temporary balloon test occlusion(BTO)of ICA.One died during the training to improve the collateral of the Willis circle.Conclusion ICA embolization is feasible for treatment of traumatic pseudoaneurysms in siphon segment of internal carotid artery after evaluating the collateral circulation of Willis circle.

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