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1.
Article in Chinese | WPRIM | ID: wpr-1014665

ABSTRACT

AIM: To investigate the regulatory effects of silybin on hepatic lipid metabolism in mice with non -alcoholic steatohepatitis (NASH) induced by high - fat and high-cholesterol (HFHD) diet. METHODS: Mice were fed a HFHD diet to construct a NASH model, and serum levels of triacylglycerol (TAG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were measured using biochemical kits. H&E staining and oil red O staining were used to detect histopathological changes in the liver. Lipidomics was used to detect the alterations of hepatic lipid metabolism in NASH mice. RESULTS: Silybin significantly inhibited the increase of body weight, liver weight and abdominal fat, decreased serum T-CHO, TAG and LDL-C levels, improved hepatic lipid droplet accumulation and ballooning degeneration, and back-regulated hepatic palmitoleic acid (C16: 1) and polyunsaturated long-chain fatty acids (PUFAs) in NASH mice. CONCLUSION: Silybin possibly reduced hepatic lipid accumulation and lipotoxicity by modulating abnormal hepatic lipid metabolism in mice induced by HFHC diet.

2.
Yao Xue Xue Bao ; (12): 3082-3089, 2023.
Article in Chinese | WPRIM | ID: wpr-999046

ABSTRACT

An LC-MS method with natural isotope abundance correction and a 1H NMR relative quantitative method were established to determine the deuterium incorporation of donafenib tosilate, a new deuterated drug molecule. First, the peak areas of isotopic impurities (non-deuterated and incompletely deuterated impurities) and deuterated drug were recorded through the single ion monitoring (SIM) mode of the established LC-MS method and then corrected in terms of the natural isotope abundance offered by ChemDraw soft, removing the nature isotope interference from 13C, 37Cl, etc. The corrected areas were subsequently used to calculate mol% of isotopologues (D0, D1, D2, D3) and Atom% D, namely, deuterium incorporation. In addition, a 1H qNMR experiment was conducted with the aromatic proton at δ 8.63 and the residual proton of isotopic impurities at δ 2.79 as quantitative peaks. The mixture of DMSO-d6 and D2O (10∶1) was employed as the solvent to change the spin-coupling between the residual proton and active hydrogen so that the residual proton could be measured as the single peak, and the sensitivity was greatly improved. The acquisition parameters were also optimized, and Atom% 1H and the deuterium incorporation were then calculated. The two methods were applied to samples of three commercial batches, and the testing results were almost consistent. Both methods proved accurate, sensitive, fast and independent of standard substances and accurate weighing, which could be applied to the determination of the deuterium incorporation of donafenib tosilate and provide a reference for other deuterated drugs.

3.
Chin. med. j ; Chin. med. j;(24): 2444-2455, 2020.
Article in English | WPRIM | ID: wpr-877859

ABSTRACT

In recent years, the research of immune checkpoint inhibitors has made a great breakthrough in lung cancer treatment. Currently, a variety of immune checkpoint inhibitors have been applied into clinical practice, including antibodies targeting the programmed cell death-1, programmed cell death-ligand 1, and cytotoxic T-lymphocyte antigen 4, and so on. However, not all patients can benefit from the treatment. Abnormal antigen presentation, functional gene mutation, tumor microenvironment, and other factors can lead to primary or secondary resistance. In this paper, we reviewed the molecular mechanism of immune checkpoint inhibitor resistance and various combination strategies to overcome resistance, in order to expand the beneficial population and enable precision medicine.


Subject(s)
Humans , B7-H1 Antigen , CTLA-4 Antigen , Drug Resistance , Immune Checkpoint Inhibitors , Immunotherapy , Lung Neoplasms/drug therapy , Tumor Microenvironment
4.
Article in Chinese | WPRIM | ID: wpr-743230

ABSTRACT

Objective To explore the value of trauma-care check list (TCC) and quick sequential organ failure assessment (qSOFA) on the early diagnosis of severe trauma with sepsis,and analyze the treatment time lines.Methods Totally 120 patients with severe trauma treated in Taizhou People's Hospital from February 2017 to January 2018 were reviewed.Sixty cases adopted TCC and qSOFA trauma care integration process (integration group),and the rest 60 cases adopted systemic inflammatory response syndrome (SIRS) score and emergency surgery multi-section support process (traditional group).According to the 2016 International Sepsis Guide Criteria,the diagnostic sensitivity and specific degrees of the two groups were calculated.The treatment time node,blood loss,complication rate,postoperative survival rate,and the total length of hospital stay of the two groups were analyzed.Results Of the 60 cases in the integration group,32 cases were confirmed severe trauma with sepsis,and 27 cases were confirmed in 41 primary diagnosed patients,with a diagnostic sensitivity of 84.38% and a specific degree of 50.00%.In the traditional group,30 cases were confirmed severe trauma with sepsis,and 25 cases were confirmed in 38 primary diagnosed patients with a diagnostic sensitivity of 83.33% and a specific degree of 56.67%.The significant shorter MDT consultation time,primary diagnosis time of sepsis,the duration from injury to surgery time and total hospitalization time were statistically significant different between the two groups (P<0.05).Patients in the integration group had significantly lower incidence of postoperative complications and 28-day fatality rate,but there was no significant difference between them (P>0.05).Conclusions TCC and qSOFA score in the treatment of severe trauma can optimize salvage process,significantly shorten the treatment time,and reduce postoperative complications.Moreover,qSOFA score and SIRS score have the same effect on the early diagnosis of sepsis in patients with severe trauma.

5.
Article in Chinese | WPRIM | ID: wpr-497720

ABSTRACT

Objective To investigate the role of intestinal fatty acid binding protein (IFABP) and blood procalcitonin (PCT) in diagnosis of traumatic Intestinal rupture in early stage.Methods The clinical data of 58 patients with abdominal injuries admitted from May 2012 to April 2016 were retrospectively analyzed.All 58 patients were divided into intestinal rupture group (n =21) and nonintestinal rupture group (n =37).The concentrations of IFABP and PCT were detected,analyzed and compared between two groups at different intervals.Results The IFABP and PCT in intestinal rupture group were significantly higher than those in non-intestinal rupture group.The IFABP and PCT in intestinal rupture group significantly decreased after operations.There were significantly differences in IFABP and PCT between two groups at admission,4 hours after admission,preoperative period,and 24 hours after operation.However,these differences disappeared at 72 hours after operation.At the same time,the accuracy rate 92.4%,sensitivity 96.3%,specificity 72.8% found in combination of these two biomarkers were significantly higher than those of IFABP and PCT measured separately.Conclusions The combination of IFABP and PCT detection can be used as an indicator for the diagnosis of traumatic intestinal rupture in the early stage.

6.
Chinese Pharmacological Bulletin ; (12): 465-469, 2015.
Article in Chinese | WPRIM | ID: wpr-465655

ABSTRACT

The degranulation of mast cells represents a pivotal e-vent in the allergic disorders.The Src family kinases(SFKs)are as a starting signal in the activation of mast cell.Lyn,Fyn and Syk play important regulatory role in the degranulation of mast cells.Regulating SFKs can reduce the degranlation process and inhibit the allergic disorders.Therefore,SFKs inhibitors can be potential drugs in the allergy.It is necessary to study the targeted medicine of SFKs,which will be a new direction of drug develop-ment.

7.
Article in Chinese | WPRIM | ID: wpr-239415

ABSTRACT

<p><b>OBJECTIVE</b>To study the cell morphology change in dormancy and proliferation stage of colorectal cancer stem cells in order to provide reference to the treatment of colorectal cancer.</p><p><b>METHODS</b>The subpopulation of EpCAM(high)/CD44(+)/CD133(+) was isolated from fresh colorectal cancer tissues. These cells were tested by xenograft assay in NOD/SCID nude mice. Colorectal cancer stem cells underwent three-dimensional culture, and the growth curve of stem cells was drawn by WST-1. The expression of P27 and Ki-67 was examined by flow cytometry to understand the phase of dormancy and proliferation of colorectal cancer stem cells. Then the morphological differences of colorectal cancer stem cells between dormant and proliferation stages were recognized by immunofluorescence staining of actin.</p><p><b>RESULTS</b>The percentage of EpCAM(high)/CD44(+)/CD133(+) was 1.6%, and the subpopulation was confirmed to be colorectal cancer stem cells by means of the experiment of tumorigenicity in vivo. The growth curve of colorectal cancer stem cells was "S" type. Colorectal cancer stem cells grew slowly in the first three days. The expression of P27 was gradually up-regulated, and the level of Ki-67 was very low. These cells remained quiescence, which was the so-called dormancy. The expression of Ki-67 of colorectal cancer stem cells was at high level since the fourth day, and the P27 level was very low. According to the growth curve, this period belonged to the proliferative stage of colorectal cancer stem cells. On immunofluorescence staining, colorectal cancer stem cells with high level of P27 were round, large, and few pseudopodium, but no obvious death was found. These cells showed characteristics of dormancy. In contrast, the stem cells with high level of Ki-67 had much pseudopodium, showing proliferation and invasion.</p><p><b>CONCLUSIONS</b>Cancer recurrence and metastasis may be associated with the change of growth state of cancer stem cells. Colorectal cancer stem cells in the proliferation stage show greater proliferative and invasive ability as compared to the dormancy stage, which provides a new perspective for the treatment of colorectal cancer, and recurrence and metastasis of other tumors.</p>


Subject(s)
Animals , Mice , Antigens, CD , Antigens, Neoplasm , Cell Adhesion Molecules , Cell Cycle Checkpoints , Cell Proliferation , Cell Shape , Colorectal Neoplasms , Epithelial Cell Adhesion Molecule , Glycoproteins , Mice, Inbred NOD , Mice, Nude , Mice, SCID , Neoplasm Recurrence, Local , Neoplastic Stem Cells , Cell Biology
8.
Article in Chinese | WPRIM | ID: wpr-447651

ABSTRACT

Objective To investigate the utilization of damage control surgery (DCS) in treatment of the severe abdominal traumas.Methods The clinical data of 56 patients with severe abdominal traumas treated by DCS (Emergency operation,fluid resuscitation and definitive operation were used gradually.)from January 2011 to January 2013 in Department of General Surgery,Taizhou People's Hospital were retrospectively analyzed.Results Of the 56 patients,52 were cured and discharged from hospital,the clinical curative rate was 92.9%; 4 patients dead.Conclusions Abdominal traumas under going operations by applying the damage control surgery significantly increased the clinical curative rate.

9.
Yao Xue Xue Bao ; (12): 1817-1822, 2013.
Article in Chinese | WPRIM | ID: wpr-298005

ABSTRACT

Naringin has been reported to possess a wild range of biological activities. However, the route and metabolites of naringin produced by intestinal bacteria are not well understood. In this paper, different bacteria were isolated from human feces and their abilities to convert naringin to different metabolites were studied. Ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) with automated data analysis software (MetaboLynx) was applied to fast analysis of naringin metabolites. Using MSE and mass defect filter techniques, three metabolites were detected and tentatively identified. The results indicated that acetylation, hydrolyzation and hydrolyzation with hydrogenation were the major metabolic pathways of naringin in vitro. Then, we studied the gene sequence of the 16S rRNA of the bacteria by extraction of genomic DNA of the strain, PCR amplification and clone of the 16S rRNA. The consequence proved that Enterococcus sp.30, Bacillus sp.46, Escherichia sp.54 and Escherichia sp.63 have the peculiar metabolism characteristic of naringin.


Subject(s)
Female , Humans , Bacillus , Genetics , Metabolism , Chromatography, High Pressure Liquid , Enterococcus , Genetics , Metabolism , Escherichia , Genetics , Metabolism , Feces , Microbiology , Flavanones , Metabolism , Metabolic Networks and Pathways , Phylogeny , RNA, Ribosomal, 16S , Genetics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
10.
Article in Chinese | WPRIM | ID: wpr-435023

ABSTRACT

Objective To investigate the role of Gly82Ser polymorphism of receptor for advanced glycation end-products (RAGE) in the genesis and progression of colon cancer in Chinese population.Methods Using the method of PCR-restriction fragment length polymorphism (PCR-RFLP),the Gly82Ser genotype of RAGE were examined in 90 colon cancer patients and 78 control subjects age and sex matched.Analyses stratified by TNM and tumor differentiation were conducted to check the associations of the Gly82Ser gene polymorphisms in RAGE and development of colon cancer.Results The genotype distribution was in agreement with that predicted under Hardy-Weinberg equilibrium for both patients and controls (both P > 0.05).With the GG genotype as reference,the odds ratio (OR) for heterozygous GG and carriers with S allele (GS and SS) were 2.037 (95% CI:1.207-3.438) and 2.022 (95% CI:1.275-3.208),respectively,which had a significantly higher risk of colon cancer.Moreover,the elevated colon cancer risk was especially evident in patients with TNM (Ⅲ + Ⅳ) and/or patients with poor differentiation by stratification analysis (OR,3.575,95% CI:1.495-8.550 and OR,3.580,95% CI:1.390-9.217,respectively).Conclusions The RAGE Gly82Ser polymorphism may confer not only an increased risk of colon cancer but also with invasion of colon cancer in the Chinese population.

11.
Zhongguo dangdai erke zazhi ; Zhongguo dangdai erke zazhi;(12): 866-869, 2013.
Article in Chinese | WPRIM | ID: wpr-345690

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of sleep deprivation on intelligence development in primary school students.</p><p><b>METHODS</b>Between June 2009 and April 2010, 316 grade 5 students aged 10-11 years were selected from four primary schools in four administrative districts of Changsha, China by stratified random sampling. The intelligence characteristics of children with varying degrees of sleep deprivation were investigated using the Chinese Wechsler Intelligence Scale for Children.</p><p><b>RESULTS</b>A total of 286 valid questionnaires were received, with a response rate of 90.5%. The survey was comprised of a sleep deprivation group (sleep time <8 hours per night; n=180) and a control group (sleep time ≥8 hours per night; n=106). The sleep deprivation group had significantly lower subtest scores, verbal intelligence quotient (IQ) (VIQ), performance IQ (PIQ) and full scale IQ (P<0.05) and significantly lower verbal comprehension factor score and memory/attention factor score compared with the control group (P<0.05). Compared with the control group, the moderate sleep deprivation subgroup had significantly decreased VIQ and full scale IQ as well as verbal comprehension factor score and memory/attention factor score (P<0.05), and the severe sleep deprivation subgroup showed decreases in all scores (P<0.05). The sleep deprivation group and moderate and severe sleep deprivation subgroups had significantly higher proportions of children with VIQ-PIQ imbalance than the control group.</p><p><b>CONCLUSIONS</b>Sleep deprivation adversely affects intelligence development, especially VIQ, in primary school students, and the adverse effects of sleep deprivation are mainly seen in students with moderate and severe sleep deprivation.</p>


Subject(s)
Child , Female , Humans , Male , Intelligence , Sleep Deprivation , Psychology
12.
Zhongguo Zhong Yao Za Zhi ; (24): 2140-2146, 2013.
Article in Chinese | WPRIM | ID: wpr-346426

ABSTRACT

To provide a scientific evidence for the initial primary processing method, an ultra-high performance liquid chromatography combined with a triple quadrupole electrospray tandem mass spectrometry (UPLC-MS/MS) was used to analyze the contents variation of catechins, flavonoids, flavonoid glycosides, biflavones, terpene lactones and phenolic acids during the process of drying in the sun, in the shade, and baked with 35, 45, 60, 80 degrees C, respectively. The results show that drying in the 80 degrees C is conducive to the accumulation of catechins, flavonoid glycosides, terpene lactones, better than the effects of other procedures. Therefore, the fast drying at 80 degrees C is beneficial for the retention of various types of active ingredient of Ginkgo biloba, and this method could be applied as a preferably dry processing.


Subject(s)
Chromatography, High Pressure Liquid , Ginkgo biloba , Chemistry , Plant Leaves , Chemistry , Tandem Mass Spectrometry , Technology, Pharmaceutical
13.
Zhongguo Zhong Yao Za Zhi ; (24): 376-380, 2013.
Article in Chinese | WPRIM | ID: wpr-346813

ABSTRACT

<p><b>OBJECTIVE</b>To establish an ultra-high performance liquid chromatography coupled with triple quadrupole mass (UPLC-TQ-MS) for determination of four terpene lactones.</p><p><b>METHOD</b>Chromatographic separation was carried out on a ACQUITY UPLC BEH C18 column (2.1 mm x 100 mm, 1.7 microm) with isocratic elution of 70% methanol at a flow rate of 0.4 mL x min(-1), the column temperature was set at 30 degrees C; Waters Xevo TQ worked in multiple reaction monitoring mode.</p><p><b>RESULT</b>All calibration curves were linear (r > 0.990 3) over the tested ranges. The average recoveries ranged from 98.83% to 103.9% with RSD value below 3.0%. The contents of total terpene lactones in Ginkgo biloba leaves were significantly different in different ages. The contents in the leaves of young ginkgo tree were higher than that in old tree.</p><p><b>CONCLUSION</b>The method was simple and fast with high precision, sensitivity and repeatability, which can be used for qualitative and quantitative analysis of terpene lactones in G. biloba leaves.</p>


Subject(s)
Calibration , Chromatography, High Pressure Liquid , Methods , Ginkgo biloba , Chemistry , Lactones , Mass Spectrometry , Methods , Plant Leaves , Chemistry , Reproducibility of Results , Terpenes , Time Factors
14.
Yao Xue Xue Bao ; (12): 1043-1048, 2012.
Article in Chinese | WPRIM | ID: wpr-276202

ABSTRACT

By investigating the interaction between components from Flos Genkwa (FG) and Radix et Rhizoma Glycyrrhizae (RRG) and the dissolution profile of toxic components in co-decoction, the characteristics and possible mechanism of incompatibility were revealed. Ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) and ultra-high performance liquid chromatography triple-quadrupole mass spectrometry (UPLC-TQ/MS) were used to analyze multi-components in different herb extractions prepared by different ratios of FG/FG processed by vinegar (FGV) and RRG, which reflect the interaction and characteristics of multiple components in incompatibility combinations. The results showed that the components dissolution was influenced by compatibility ratio with certain regularity. Whether FG processed by vinegar or not, with the increase of RRG in co-decoction, the dissolution of diterpenes, especially for yuanhuacine, yuanhuadine and yuanhuajine, the toxic ingredients of FG, increased significantly. From these results, the material basis and one possible mechanism of incompatibility between FG and RRG is the increasing dissolution of diterpenes, toxic components of FG in co-decoction process, which caused by interaction between multi-components in these two herbs.


Subject(s)
Acetic Acid , Chemistry , Chromatography, High Pressure Liquid , Daphne , Chemistry , Diterpenes , Drug Incompatibility , Flowers , Chemistry , Glycyrrhiza uralensis , Chemistry , Plant Roots , Chemistry , Plants, Medicinal , Chemistry , Rhizome , Chemistry , Spectrometry, Mass, Electrospray Ionization , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Tandem Mass Spectrometry , Terpenes
15.
Zhongguo Zhong Yao Za Zhi ; (24): 3256-3263, 2012.
Article in Chinese | WPRIM | ID: wpr-308605

ABSTRACT

<p><b>UNLABELLED</b>To establish a rapid component identification method for chemical constituents contained in traditional Chinese medicines and natural products by using liquid chromatography-mass spectrometry database technology.</p><p><b>METHOD</b>Ultra-high performance liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was applied to collect the retention time and mass spectrum data of chemical constituents contained in traditional Chinese medicines and natural products, and establish a mass spectrum database. The Identify function from Chromalynx XS module in Masslynx workstation was applied to identifiy each chromatographic peak in an automatic and rapid manner. The retention time correction method was used to correct the retention time of identical compounds in different chromatographic systems, in order to improve the adaptability of the method.</p><p><b>RESULT</b>Currently, the UPLC-MS library contains LC-MS information of more than 210 such chemical constituents as alkaloids, flavonoids, quinones, phenylpropanoids and terpenes, including retention times, xact molecular weights of quasi-molecular ion peaks and their fragment ions under positive and negative modes, respectively. The detection rate of identifying samples by suing the method was around 90%.</p><p><b>CONCLUSION</b>Compared with manual identification, the method is simple, rapid and highly accurate. The usage of the linear correction for retention times helps solve the disunity of mass spectrum databases caused by different retention times in different chromatographic systems to a certain extent, and expand the application range of the method.</p>


Subject(s)
Chromatography, Liquid , Methods , Databases, Factual , Mass Spectrometry , Methods , Medicine, Chinese Traditional
16.
Yao Xue Xue Bao ; (12): 1374-1379, 2011.
Article in Chinese | WPRIM | ID: wpr-232980

ABSTRACT

Sinisan is a widely used traditional Chinese medicine (TCM) in treating various diseases; however, the in vivo metabolic profile of its multiple components remains unknown. In this paper, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was applied to identify the metabolites of Sinisan extract in rat plasma, urine, feces and bile after intragastric administration. Using MS(E) and mass defect filter techniques, 41 metabolites of 10 parent compounds (naringin, naringenin, hesperidin, neohesperidin, liquiritin, liquiritigenin, glycyrrhizic acid, glycyrrhetinic acid, saikosaponin a and saikosaponin d) were detected and tentatively identified. It was shown by our results that these compounds was metabolized to the forms of hydroxylation, glucuronidation, sulfation, glucuronidation with sulfation and glucuronidation with hydroxylation in vivo.


Subject(s)
Animals , Male , Rats , Administration, Oral , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Metabolism , Pharmacokinetics , Flavanones , Metabolism , Pharmacokinetics , Glucosides , Metabolism , Pharmacokinetics , Glycyrrhizic Acid , Metabolism , Pharmacokinetics , Hesperidin , Metabolism , Pharmacokinetics , Hydroxylation , Plants, Medicinal , Chemistry , Random Allocation , Rats, Sprague-Dawley , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
17.
Zhongguo dangdai erke zazhi ; Zhongguo dangdai erke zazhi;(12): 973-976, 2011.
Article in Chinese | WPRIM | ID: wpr-272418

ABSTRACT

<p><b>OBJECTIVE</b>To study the adverse effects of sleep problems and sleep insufficiency on attention in class and pupils' acadamic achievements.</p><p><b>METHODS</b>A total of 1138 students from four primary schools at ages of 6-12 years were randomly sampled from four districts of Changsha city, Hunan Province June 2009 to April 2010. The inquired items included sleep problems, sleep time, sleep quality, attention in class and academic achievements. Teachers and parents observed the pupils according to the unified requirements for 3 months and then filled out the questionnaires.</p><p><b>RESULTS</b>The total valid inquiry tables were 1091 with the answering rate of 95.87%, including 549 boys and 542 girls. The sleep quality was more poor in children with sleep problems or sleep insufficiency than in children with normal sleep. The sleep quality was reduced and aggravated along with the increasing sleep problems and the reducing sleep time. The attention in class and academic achievements in children with sleep problems or sleep insufficiency were more poor than in children with normal sleep. The sleep quality index was negatively correlated with attention in class and academic achievements. The attention in class was positively correlated with academic achievements.</p><p><b>CONCLUSIONS</b>The decline in sleep quality directly affects the attention in class and reduce the academic achievements in primary school children.</p>


Subject(s)
Child , Child, Preschool , Female , Humans , Male , Achievement , Attention , Schools , Sleep
18.
Zhonghua Yu Fang Yi Xue Za Zhi ; (12): 721-725, 2010.
Article in Chinese | WPRIM | ID: wpr-291508

ABSTRACT

<p><b>OBJECTIVE</b>To achieve secretory and extracellular production of recombinant dengue virus serotypes I-IV envelope glycoprotein domain III (DENV-1-4 EDIII) in Pichia pastoris.</p><p><b>METHODS</b>EDIII genes of DENVI-IV were amplified and cloned into vector pPIC9K, respectively. These recombinant plasmids were then linearized and transferred into Pichia pastoris strain GS115. Clones highly produced in 4.0 mg/ml G418 were amplified and induced by methanol to achieve the secreted recombinant proteins. Ni-NTA agarose beads were used for purification, while SDS-PAGE and Western blotting were used for identification.</p><p><b>RESULTS</b>The recombinant plasmids pPIC9K-DENV-1-4 EDIII were constructed and successfully transferred into Pichia pastoris strain GS115. The recombinant EDIII proteins were expressed in a secretory way with the molecular weight about 12 × 10(3) and specifically identified by anti-His monoclonal antibody and anti-DENVI-IV mice sera.</p><p><b>CONCLUSION</b>DENVI-IV EDIII proteins are successfully achieved from Pichia pastoris expression system and could be used for development of dengue vaccines, diagnostic reagents and study of biological function of the E protein.</p>


Subject(s)
Dengue Virus , Genetics , Genetic Vectors , Pichia , Metabolism , Recombinant Proteins , Genetics , Viral Envelope Proteins , Bodily Secretions
19.
Zhongguo zhenjiu ; (12): 213-216, 2009.
Article in Chinese | WPRIM | ID: wpr-257115

ABSTRACT

<p><b>OBJECTIVE</b>To probe into effective methods for treatment of ischemic cerebral infarction and specificity of acupoints in the acupuncture treatment.</p><p><b>METHODS</b>The rat model of middle cerebral artery occlusion (MCAO) was established with thread ligation according to Zea-Longa's method. They were divided into normal group, non-operation group, model control group, non-acupuncture group and acupuncture group, and the acupuncture group was divided into sham-acupuncture group and four Xingnao Kaiqiao groups (contain Shuigou group, Neiguan group, Chize group, Sanyinjiao group and Weizhong group). Then they were treated by acupuncture at "Shuigou" (GV 26), "Neiguan" (PC 6), "Chize" (LU 5), "Sanyinjiao" (SP 6), "Weizhong" (BL 40) and non-acupoints, 3 times/second, for 5 seconds. Cerebral blood blow (CBF) was used for assessment of the effect.</p><p><b>RESULTS</b>As compared with the model control group, in the non-acupuncture group CBF did not significantly change (P>0.05); compared with the non-acupuncture group, after acupuncture CBF was significantly increased in the Shuigou group and the Neiguan groups (all P<0.05), but did not significantly increased in the Chize group, Sanyinjiao group and Weizhong group (all P>0.05).</p><p><b>CONCLUSION</b>CBF has a tendency of spontaneous cure within 72 h after cerebral infarction in the MCAO rat; acupuncture can significantly improve CBF in the MCAO rat, so it is an effective method for treatment of ischemic stroke; among the acupoints in the "Xingnao Kaiqiao" needling method, "Shuigou" (GV 26) and "Neiguan" (PC 6) have obvious effect in improvement of CBF, indicating acupoint specificity of "Shuigou" (GV 26) and "Neiguan" (PC 6) in treatment of ischemic stroke.</p>


Subject(s)
Animals , Humans , Male , Rats , Acupuncture Points , Acupuncture Therapy , Brain , Brain Ischemia , Therapeutics , Cerebral Infarction , Therapeutics , Disease Models, Animal , Random Allocation , Rats, Wistar
20.
Zhongguo zhenjiu ; (12): 733-738, 2009.
Article in Chinese | WPRIM | ID: wpr-257915

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of acupoint Shuigou (GV 26) and non-acupoint on inhibiting the neuronal necrosis induced by the middle cerebral artery obstruction (MCAO) in rats through the morphological observation.</p><p><b>METHODS</b>Forty two healthy adult male Wistar rats were randomly divided into a normal group, false-ope ration group, model control group, non-acupuncture group, Shuigou-acupuncture group and non-acupoint acupuncture group, 7 rats in each group. Besides the normal and false-operation groups, the experimental modal of MCAO was established in the other groups by using the Zea-Longa thread method. Acupoint Shuigou (GV 26) and non-acupoint (below the costal region) were selected respectively in the Shuigou-acupuncture group and non-acupoint acupuncture group for puncturing at 180 times/min for 5 seconds. Its effect was estimated by measuring the rate of the neuronal necrosis at the level of the light microscope and the degree of the neuronal necrosis at the level of the electron microscope.</p><p><b>RESULTS</b>(1) At the level of the light microscope, comparing with the model control group (0.66 +/- 0.18), of the neuronal necrosis there were no significant differences in the rate of neuronal necrosis in the non-acupuncture group (0.67 +/- 0.34) and non-acupoint acupuncture group (0.59 +/- 0.11) (both P > 0.05), while it was significantly decreased in the Shuigou-acupuncture group (0.200 +/- 12) (P < 0.05). It indicates that no intervention and wrong (non-acupoint) intervention could light the damage of the neurons, however, only right intervention (Shuigou, GV 26) can obviously decrease the rate of the neuronal necrosis, showing with the specificity of acupoint. (2) At the level of the electron microscope, the ultrastructure of the involved neurons in the non-acupuncture group and non-acupoint acupuncture group is similar to that of the model control group, showing with the serious edema and structure damage. In contrast, the ultrastructure of the involved neurons in the Shuigou-acupuncture group is similar to that of the normal group and false-operation group, showing with light neuronal damage.</p><p><b>CONCLUSION</b>Acupuncturing Shuigou (GV 26) could protect the involved neurons and inhibit the neuronal necrosis induced by the MCAO in rats, but acupuncturing the non-acupoint couldn't, which imply that there is the specificity of acupoint in the acupuncture.</p>


Subject(s)
Animals , Humans , Male , Rats , Acupuncture Points , Acupuncture Therapy , Brain Ischemia , Pathology , Therapeutics , Disease Models, Animal , Necrosis , Neurons , Pathology , Random Allocation , Rats, Wistar
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