1.Effects of compound Daqiqi decoction combined cisplatin on Bcl-2/Bax expression of nude mice ovarian cancer subcutaneous transplanted tumor.
Yang-bojun YANG ; Shuang WANG ; Shu LI ; Rong CHEN
China Journal of Chinese Materia Medica 2015;40(8):1575-1579
OBJECTIVETo determine the compound Daqiqi decoction( CDQD) combined cisplatin plays on the cell apoptosis of subcutaneous transplanted ovarian cancer in nude mice, to provide theory evidence for clinical treatment.
METHODMaking the models of subcutaneous transplanted ovarian cancer in nude mice, and divide the 40 mice with tumor into 5 groups (n = 8), the model control group, CDQD low dosed group, CDQD high dosed group, cisplatin group, cisplatin combined CDQD group, killing all the mice after 3 weeks' treatment and stripping tumors. Measure the volume and weight of the tumor and calculate tumor growth curve damps, the inhibition rate. Examining the expression of the Bcl-2(B-cell leukemia /lymphoma 2) and Bax (Bcl-2 associated x protein) mRNA and protein by the RT-PCR and the Western blot.
RESULT(O)The tumor weight shows that there was certain lighter effect in each CDQD group, and compared with cisplatin groups has no statistical significance, the cisplatin combined CDQD group is obviously lighter than that of the other group(P <0.01). The tumor growth curve damps shows that compared with model control group, the treatment group tumors had some extent narrowing (P < 0.01). (2) RT-PCR results: Bax (Bcl-2 associated x protein) mRNA expression shows that compared with model control group, the treatment group has increased (P < 0.01), and the cisplatin combined CDQD group compared with the other group is the highest, there was significant difference with the rest of the treatment group (P < 0.01). In the Bcl-2 mRNA express lowest in cisplatin combined CDQD group (P < 0.01), there has no difference between CDQD high dose group and cisplatin group. (Western blot shows: compared with model control group, the Bax protein of treatment group has increased expression (P < 0.01), the expression of Bax protein in cisplatin combined CDQD group is the highest(P < 0.01). There has no difference between CDQD high dose group and cisplatin group. The Bcl-2 protein expression of the cisplatin combined CDQD group is the lowest (P < 0.01), there has no difference between CDQD high dose group and cisplatin group.
CONCLUSIONThe effect of CDQD on subcutaneous transplantation ovarian tumor has promoting apoptosis function, its mechanism may be related to downgrade the Bcl-2 expression, higher expression of bax, stimulation on the apoptosis of tumor cells; cisplatin combined CDQD have synergistic effect.
Animals ; Cell Line, Tumor ; Cisplatin ; administration & dosage ; Disease Models, Animal ; Drug Therapy, Combination ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Ovarian Neoplasms ; drug therapy ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism
2.Effect of curcumin on the learning, memory and hippocampal Ca+/CaMK II level in senescence-accelerated mice.
Chen-you SUN ; Shuang-shuang QI ; Shu-hong SUN
Chinese Journal of Integrated Traditional and Western Medicine 2011;31(3):376-380
OBJECTIVETo explore effect of curcumin in different concentrations on learning and memory of senescence-accelerated mice (SAM) and their possible mechanisms.
METHODSMice were randomly divided into six groups: the SAMR1 normal control group, the SAMP8 model control group, the SAMP8 + solvent (the peanut oil) control group, SAMP8 + low, middle and high dose curcumin groups. Mice were gastrogavage for 25 successive days. On the next day of ending the experiment, changes of learning and memory in mice of each group were observed by Morris water maze. The hippocampal [Ca2+] was determined. Expressions of hippocampal calmodulin-dependent protein kinase II (CaMK II) and Calmodulin (CaM) mRNA were detected using Western blot and reverse transcription polymerase chain reaction (RT-PCR) respectively.
RESULTSThe latency to find the hidden platform was remarkably prolonged, the hippocampal [Ca2+]i was markedly increased, the expression of CaMK II in the hippocampal membrane and the level of hippocampal CaM mRNA were significantly reduced in the SAMP8-model control group (P < 0.05, P < 0.01). The latency to find the hidden platform was remarkably shortened in the SAMP8 + middle dose curcumin and the SAMP8 + high dose curcumin groups (P < 0.01). The hippocampal [Ca2+]i was markedly lowered, the expression of CaMK II in the hippocampal membrane and the level of hippocampal CaM mRNA obviously increased in the SAMP8 + low, middle and high dose curcumin groups (P < 0.05, P < 0.01).
CONCLUSIONCurcumin could improve learning and memory Ca2+/capacities of SAM by lowering hippocampal [Ca2+] overload, increase the hippocampal CaM mRNA level and CaMK II expression in the hippocampal dose-dependently.
Aging ; drug effects ; metabolism ; Animals ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; metabolism ; Calmodulin ; metabolism ; Curcumin ; pharmacology ; Hippocampus ; drug effects ; metabolism ; Maze Learning ; drug effects ; Memory ; drug effects ; Mice ; RNA, Messenger ; genetics
3.Characterization and subcellular localization of two SBP genes and their response to abiotic stress in soybean (Glycine max (L.) Merr.).
Yan YANG ; Shuang WANG ; Liyan HUANG ; Hongyu MA ; Yingjie SHU ; Xiaoling HE ; Hao MA
Chinese Journal of Biotechnology 2014;30(11):1709-1719
High temperature and humidity stress during seed growth and development of spring soybean can result in seed deterioration in South China. We isolated two genes (GmSBP and GmSBPL) encoding putative SBP proteins from soybean (Glycine max (L.) Merr.) to study their biological functions and response to abiotic stress,. The two SBP proteins are hydrophilic and incomplete membrane ones. Real-time quantitative (RT-PCR) analysis reveals that the expression of the two genes in the developing seeds of the seed deterioration resistant cultivar Xiangdou No. 3 and sensitive cultivar Ningzhen No. 1 was significantly affected by high temperature and humidity treatment. Meanwhile, the levels of sucrose and soluble sugar in the developing seeds of both cultivars were also affected under high temperature and humidity stress. During seed growth and development, the expression of the two genes as well as the levels of sucrose and soluble sugar reached the highest at 30 days after flower. GmSBP2 and GmSBPL were found to be differentially expressed in different soybean tissues. Sub-cellular localization indicated that two genes were located in cytoplasm and cell membrane. Our results indicate that GmSBP2 and GmSBPL might be involved in the response to abiotic stress, which will enrich our understanding of pre-harvest seed deterioration and resistance in soybean from one side.
China
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Gene Expression Regulation, Plant
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Genes, Plant
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Membrane Transport Proteins
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genetics
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Plant Lectins
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genetics
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Real-Time Polymerase Chain Reaction
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Seeds
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Soybean Proteins
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genetics
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Soybeans
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genetics
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Stress, Physiological
4.Relieving pre-exam anxiety syndrome with wrist-ankle acupuncture: a randomized controlled trial.
Shi SHU ; Tongming LI ; Fanfu FANG ; Houluo HE ; Qinghui ZHOU ; Wei GU ; Shuang ZHOU
Journal of Integrative Medicine 2011;9(6):605-610
Background: Pre-exam anxiety syndrome is a common condition occurring in pre-exam students and directly affects their examination performance and physical state. Wrist-ankle acupuncture has significant therapeutic effects in treating mental disorders and may also relieve the symptoms of pre-exam anxiety syndrome. Objective: To assess the therapeutic effect of wrist-ankle acupuncture on pre-exam anxiety syndrome. Design, setting, participants and interventions: A total of 60 students who met the inclusion criteria of pre-exam anxiety syndrome were enrolled from a university in Shanghai and they were randomly divided into treatment group and control group. There were 30 cases in each group, and no case failed to follow-up. In the treatment group, wrist-ankle acupuncture was adopted to point upper 1 bilaterally (impression between flexor carpi ulnaris tendon and ulnar margin), and there was no requirement for Deqi (arrival of qi). In the control group, sham acupuncture was adopted. The treatment was applied 3 times totally in both groups one week before the exam, once every other day, each time with the needles retained for 30 min. Main outcome measures: The therapeutic effects were compared between two groups. Before and after 3 treatments, Sarason Test Anxiety Scale (TAS) and Expectation and Treatment Credibility Scale (ETCS) were measured and evaluated. Results: The therapeutic effect experienced by the treatment group was better than that of the control group (P<0.05). There were no statistically significant differences in TAS and ETCS before treatment between the two groups. The scores of TAS after treatment in two groups were higher than those before treatment (P<0.05, P<0.01). There were statistical differences in TAS absolute difference and TAS relative difference between the two groups and the treatment group had better results (P<0.05, P<0.01). After treatment, patients in the treatment group had higher scores in ETCS than those in the control group (P<0.05, P<0.01). No adverse reaction was reported. Conclusion: Wrist-ankle acupuncture can relieve the symptoms of pre-exam anxiety syndrome significantly, and this therapy is highly safe.
5.Analysis of the data for inpatients with acute organophosphorous pesticide poisoning in Wucheng.
Yun-he HUO ; Xiu-hua WANG ; Xiao-ying SHANG ; Shuang-lian LIU ; Guang-shu CUI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2004;22(1):32-32
Acute Disease
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Adolescent
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Adult
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Aged
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Child
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Child, Preschool
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China
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Humans
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Infant
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Inpatients
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Insecticides
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poisoning
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Middle Aged
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Organophosphorus Compounds
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Pesticides
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poisoning
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Poisoning
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etiology
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mortality
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therapy
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Risk Factors
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Survival Rate
6.Chemical constituents from Euphorbia lunulata.
Ming ZHAO ; Shuang WU ; Jun LI ; Wan-Xia TANG ; Jin-Lan WANG ; Shu-Jun ZHANG
China Journal of Chinese Materia Medica 2014;39(12):2289-2294
The chemical constituents from Euphorbia lunulata was investigated in this paper. Fourteen compounds were isolated and purified by column chromatographies on silica gel and preparative HPLC. Their structures were identified by physiochemical properties and NMR data analysis as lupeol (1), euphol (2), cassipourol(3) , 24-methylenecycloartan-3beta-ol (4), 24-hydroperoxycycloart-25-en-3beta-ol (5), 25-hydroperoxycycloart-23-en-3beta-ol (6), betulin (7), uvaol (8), (23E) -25-methoxycycloart-23-en-3beta-ol (9), (23E) -cycloart-23,25-dien-3beta-ol (10), 24-methylenecycloartan-3beta, 28-diol (11), salicinolide (12), 2alpha, 3beta, 5alpha, 9alpha, 15beta-pentaacetoxy-11,12-epoxy-7beta, 8alpha-diisobutyryloxyjatropha-6 (17) -en-14-one (13) and 3beta, 5alpha, 15beta-triacetoxy-7beta-isobutyryloxy-9alpha-nicotinoyloxyjatropha-6 (17), 11(E)-dien-14-one (14). Among them, compounds 1-11 were isolated from E. lunulata for the first time.
Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Euphorbia
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chemistry
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Magnetic Resonance Spectroscopy
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Molecular Structure
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Spectrometry, Mass, Electrospray Ionization
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Stereoisomerism
7.Expression of TLR9 mRNA in peripheral blood mononuclear cells of patients with acute pancreatitis
Yujian ZENG ; Shuang LIU ; Liang SUN ; Ruo SHU ; Chengmin SHI ; Shujing GUO ; Kunhua WANG ; Huayou LUO
Chongqing Medicine 2015;(5):623-625
Objective To investigate the expression of TLR9 mRNA of peripheral blood mononuclear cells (PBMCs) in patients with acute pancreatitis .Methods Fifty two AP patients with the disease duration in 24 h were collected ,peripheral EDTAK2 coag‐ulation vein blood were collected on the first ,third and fifth day ,then plasma were cryop reserved to detect pancreatic elastase , proinflammatory cytokines and anti‐inflammatory cytokines .Then the peripheral EDTAK2 coagulation vein blood two to three months after treatment were collected in the same method to undertake these tests ,and act as the reference level value .Peripheral blood was collected from 36 acute pancreatitis patients .Three months later ,peripheral blood was collected again from these 36 peo‐ple as controls .PBMCs were isolated by Ficoll‐Hypaque gradient centrifugation .RT‐PCR was adopted to determine the relative con‐tent of the expression of TLR9 mRNA of the PBMCs .Results The relative content of expression of TLR9 mRNA were significant‐ly up‐regulated in the patients with acute pancreatitis ,compared with that of controls (P<0 .05) .The up‐regulated expression of TLR9 mRNA was related to expression of TNF‐a and IL‐1 .Conclusion The up‐regulated expression of TLR9 mRNA in acute pan‐creatitis patients indicates that infective factors might be mediated by TLR 9 .
8.The value of anti-SUMO antibody in the clinical diagnosis and treatment of primary biliary cholangitis
Shuang QI ; Lixin SHU ; Chongxu HAN
International Journal of Laboratory Medicine 2019;40(3):298-303,307
Objective In this study, we constructed a plasmid, specifically expressed SUMO protein, and to study the expression level of anti-SUMO antibody in the serum of patients with primary biliary cholangitis, systemic lupus erythematosus, rheumatoid arthritis, Connective tissue disease, sjogren′s syndrome which were typical of autoimmune diseases.And to investigate whether anti-SUMO antibodies can be used as specific serum markers of PBC.Methods Plasmids containing SUMO1, SUMO2 and SUMO3 fragments were prepared by PCR and ET cloning and introduced into E.coli for protein induction and purification, respectively.Dot blot was used to preliminarily screen anti-SUMO antibody-positive specimens from serum samples of PBC patients and were verified by western blot to obtain positive reference serum.Through the establishment of the optimal anti-SUMO antibody ELISA diagnostic system, the positive rates of three subtypes of anti-SUMO antibody in PBC, SLE, SS, RA and CTD were detected, and their differences were analyzed by chi-square test.ResultsThe anti-SUMO antibody label has a specificity of up to 99%in PBC and a sensitivity of around 86%.After chi-square test analysis, the positive detection rate of anti-SUMO antibody in PBC was higher than that of nonPBC autoimmune disease and healthy controls (P<0.01).There was no significant difference in the expression of anti-SUMO antibody in other autoimmune disease populations (non-PBC autoimmune diseases) (P>0.05).Conclusion The highly specific anti-SUMO antibody is expected to become a novel antibody for diagnosis of PBC, which is of great significance for improving the clinical diagnosis efficiency of PBC.
9.The effect of vagus nerve stimulation on the prognosis of rats after cardiopulmonary resuscitation under different treatment timings
Weijing SHAO ; Shuang XU ; He HUANG ; Tingting SHU ; Yuhan ZHANG ; Licai LIANG ; Yuran ZHOU ; Peng SUN
Chinese Journal of Emergency Medicine 2021;30(1):31-36
Objective:To explore the protective effect of vagus nerve stimulation (VNS) on the prognosis of rats suffering from cardiac arrest/cardiopulmonary resuscitation (CA/CPR) under different treatment timings.Methods:The method of percutaneous epicardial electrical stimulation was used to establish CA model of rat. Fifty-three male SD rats were randomly (random number) divided into the sham group ( n=5), CPR group ( n=12), PRE group ( n=12), POST5 group ( n=12) and POST30 group ( n=12). The sham group did not experience CA/CPR. VNS treatment was started at 30 min before CA (PRE group, n=12), 5 min after recovery of spontaneous circulation (ROSC) (POST5 group, n=12), and 30 min after ROSC (POST30 group, n=12) in different VNS-treated group, respectively. The electrical stimulation was applied to the vagus nerve for 30 min with a unified parameter. The neurological deficit scores at 24, 48, and 72 h after ROSC were recorded, and the survival rate in each group was observed. TUNEL staining was used to detect the apoptosis of cortical area and the expression of α7 nicotinic acetylcholine receptor (α7nAChR) in brain tissue was measured by immunofluorescence at 72 h after ROSC. Variables were compared with one-way analysis of variance, and survival for Kaplan-Meier curves were tested with the log-rank test. A P value less than 0.05 was considered statistically significant. Results:Compared with the CPR group (survival rate 33.33%), both pre-treatment (survival rate 75%) and post-treatment of VNS (POST5 group survival rate 75% and POST30 group survival rate 83.33%) significantly improved the 72 h survival rate after CPR ( P<0.05), mitigated neurological deficits after ROSC, reduced the positive rate of apoptosis neurons, and up-regulated the expression of α7nAChR in cerebral cortex. There was no significant difference among the VNS-treated groups (all P>0.05). Conclusions:Both pre-treatment and post-treatment of VNS can play a protective role in rats after CA/CPR, which may be related to the activation of α7nAChR-mediated anti-inflammatory and anti-apoptosis effects.
10.Development of a Forensic Multiplex Amplification STR Kit for 15 Autosomal STR Loci and 10 Y-STR Loci.
Yan DONG ; Shuang-shuang LIN ; Yu CAO ; Wei-wei WU ; Shu-qin HUANG ; Wei-guo ZHENG ; Fa-yuan LI ; Bin-wen GE ; Yu-lin GUO ; Huai-gu ZHOU
Journal of Forensic Medicine 2015;31(5):373-380
OBJECTIVE:
To establish a multiplex STR genotyping method for autosomal STR and Y-STR loci in forensic biological practice.
METHODS:
Widely used autosomal STR loci and Y-STR loci were selected. A set of PCR primers was designed, and a 5-dye fluorescent labeled STR multiplex PCR reagent kit was developed.
RESULTS:
A kit was developed which can simultaneously detect 15 autosomal STR loci, 10 Y-STR loci, and an Amelogenin.
CONCLUSION
The 15 autosomal STR plus 10 Y-STR kit in combination with capillary electrophoresis method was used to STR genotyping with accurate and reliable results. The new one-step testing kit can potentially be widely used in forensic cases and DNA databank in the future.
Amelogenin
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Chromosomes, Human, Y/genetics*
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DNA Primers
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Databases, Nucleic Acid
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Forensic Genetics/methods*
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Genotype
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Genotyping Techniques/instrumentation*
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Humans
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Indicators and Reagents
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Microsatellite Repeats
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Multiplex Polymerase Chain Reaction