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Objective:To investigate the antianxiety effect of schisandrin B (SchB) in the ICR mice,and to elucidate its related mechanisms.Methods:The ICR mice were divided into blank control group (administered with distilled water intragastrically),2.5 mg · kg-1 SchB group,5 mg · kg-1 SchB group,and 10 · mg · kg-1 SchB group (administered with SchB intragastrically).After the successive administration for 7 d,cross maze test and zero maze test were conducted in all the mice,and then the peripheral blood and brain tissue samples of the mice in blank control group and 10 mg · kg-1 SchB group were taken.The levels of γ-aminobutyric acid (GABA) and glutamic acid (Glu) in the peripheral blood and brain tissue of the mice were detected by ELISA,and the ratio of GABA/Glu was calculated.The expression levels of the α1 subunit of the γ-aminobutyric acid (GABAARα1) and the γ2 subunit of the γ-aminobutyric acid (GABAARγ2) in the brain tissue of the mice were detected by Western blotting method.Results:In the elevated plus maze test,the percentages of number of open arm entries in 5.0 and 10.0 mg · kg 1 SchB groups were significantly increased compared with blank control group (P＜0.05 or P=0.01),and the percentages of time of open arm entries were significantly increased (P=0.05 or P＜0.01).In the elevated zero maze test,the percentages of number of open arm entries of the mice in 5.0 and 10.0 mg · kg-1SchB groups were significantly increased compared with blank control group (P＜0.01),and the percentages of number of entering open arm probe were significantly increased (P＜0.01).Compared with blank control group,the GABA levels in the peripheral blood and brain tissue of the mice in 10.0 mg · kg-1 SchB group,were significantly increased (P＜0.01),the Glu levels in the peripheral blood and brain tissue were significantly reduced (P＜0.01),the ratios of GABA/Glu were significantly increased (P＜0.01),and the expression levels of GABAARα1and GABAARγ2 in the brain tissue were significantly increased (P＜0.01).Conclusion:SchB has an antianxiety effect in the mice,and the effect may be related to its regulating the levels of GABA and Glu in the peripheral blood and brain tissue and the expression levels of GABAA Rα1 and GABAA Rγ2 in the brain tissue.
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Objective:To investigate the lipid-lowering effect and antioxidant activity of polysaccharide from Schisandra Chinensis (SCP)in the rats with non-alcoholic fatty liver disease (NAFLD)induced by high-fat diet,and to provide a theoretical basis for the development and utilization of Schisandra Chinensis.Methods: A total of 32 male Wistar rats were selected.Sixteen from the 32 rats were randomly selected and divided into normal control group (intragastrical administration of water,combined with normal diet,n = 8)and SCP group (intragastrical administration of 50 mg·kg-1 SCP,combined with normal diet,n=8).The remaining 16 rats were fed with high-fat diet for 4 weeks and the confirmed NAFLD rat models were set up.A total of 16 NAFLD rats were randomly divided into NAFLD group (intragastrical administration of water, combined with high-fat diet,n = 8 ) and NAFLD+SCP group (intragastrical administration of 50 mg·kg-1 SCP,combined with high-fat diet,n=8).After treated for 12 weeks,the body weights of all the rats were weighed and the liver index was calculated.The levels of total cholesterol (TC),triglyceride (TG),low density lipoprotein cholesterol (LDL-C),high density lipoprotein cholesterol (HDL-C),alanine aminotransferase (ALT)and aspartate aminotransferase (AST)in the serum of all the rats were determined.The levels of TC and TG in liver tissue of the rats were measured by enzymatic method. The malondialdehyde (MDA)levels and superoxide dismutase (SOD)activities in serum and liver tissue and the glutathione (GSH)levels in liver tissue of the rats were analyzed by TBA,xanthinoxidase and microscale enzyme methods,respectively. HE staining was used to observe the pathomorphology of liver tissue of the rats. Results:Compared with normal control group,the liver index of the rats in NAFLD group was increased (P <0.01);the levels of TC,TG,LDL-C,ALT and AST in serum of the rats were increased (P <0.01),the levels of TC and TG in liver tissue of the rats were increased (P <0.01),the MDA level was increased (P <0.01)and the SOD activity was decreased (P <0.01),and the GSH levels in liver tissue and serum were decreased (P <0.01). Compared with NAFLD group,the body weight and liver index,serum levels of TC,TG,LDL-C ,ALT and AST of the rats in NAFLD + SCP group were decreased (P < 0.05 ),the levels of TC and TG in liver tissue were decreased (P <0.01),the MDA level was decreased (P <0.01),the SOD activities in serum and liver tissue were increased (P < 0.01),and the level of GSH in liver tissue was increased (P < 0.01).The HE staining results showed that the structure of hepatic lobules of the rats in NAFLD group was disordered and showed significant hepatic steatosis,and the hepatic steatosis in hepatic lobules of the rats in NAFLD+SCP group was significantly reduced.Conclusion:SCP has a regulation effect in the NAFLD rats induced by high-fat diet,and its mechanism may be related to the anti-oxidative stress.
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Objective:To observe the improvement of acidic polysaccharose of Schisandrae Chinensis (SCP-A) on the learning and memory functions of Alzheimer's disease (AD)model mice induced by Aβ25-35 ,and to clarify the related mechanisms. Methods:A total of 75 male C57BL mice were randomly divided into control group (injected with saline in intracerebroventricular,given distilled water intragastrically),model group (injected with Aβ25-35 intracerebroventricularly,given distilled water intragastrically),and 5,10,20 mg·kg-1 SCP-A groups (injected with Aβ25-35 intracerebroventricularly,given SCP-A intragastrically) (n = 15).The agents were administered once daily for 14 d,in which Aβ25-35 was injected intracerebroventricularly on the 8th day.On the 15th day after administration,step-through test and Morris water maze test were used to detect the escape latency,number of errors,and time of finding platform,number of passing platforms,dwell time in target quadrant of the mice;Western blotting method was used to detected the levels of Tau,GSK-3β and their phosphorylated proteins in hippocampus tissue of the mice.Results:The step-through test results showed that compared with control group, the escape latency of the mice in model group was obviously shortened and the number of errors was significantly increased (P <0.01);compared with model group,the escape latencies of the mice in 10 and 20 mg·kg-1 SCP-A groups were significantly prolonged (P <0.01),the number of errors was significantly decreased (P <0.05 or P <0.01).The Morris water maze test results showed that compared with control group,the time of finding platform of the mice in model group was significantly prolonged,the number of passing platform and dwell time in target quadrant were significantly reduced (P <0.01);compared with model group,the time of finding platform of the mice in 10 and 20 mg· kg-1 SCP-A groups was significantly shortened (P < 0.01),and the number of passing platform and the dwell time in target quadrant were significantly increased (P <0.01).The Western blotting results showed that compared with model group,the expression levels of phosphorylated protein Tau Ser199,Tau Ser396 and Tau Ser404 in hippocampus of the mice in 20 mg·kg-1 SCP-A group were significantly decreased (P <0.05), and the expression level of GSK-3βwas significantly decreased (P <0.01);the expression level of phosphorylated protein GSK-3βTyr216 was significantly decreased,and the expression level of phosphorylated protein GSK-3βSer9 was significantly increased (P <0.05).Conclusion:SCP-A has an anti-AD effect,which is related to regulating the activity of GSK-3βto reduce the level of phosphorylated protein Tau in the hippocampus tissue of the mice.
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Objective:To study the effects of compound schisandra extracts (CSE) (schisandra,astragalus,acanthopanax,and rhodiola)on the exhaustive swimming time and the levels of blood urea nitrogen(BUN),serum lactic acid(LD),liver glycogen and muscle glycogene,superoxide dismutase(SOD)activity,and malonaldehyde(MDA) level in the mice and to charify its anti-fatigue effect and the mechanism.Methods:Eighty male ICR mice were randomly divided into blank control group,50 mg·kg-1 CSE group,100 mg·kg-1 CSE group,and 200 mg·kg-1CSE group;there were 20 mice in each group.The mice were administered orally for 30 d.Then 10 mice were randomly selected for exhaustive swimming test in each group and the exhaustive swimming time of the mice was recorded.The remaining 10 mice in each group were used for 90 min swimming,then all the mice were sacrificed and the blood and tissue samples were taken for the measurement of the levels of BUN,LD,liver glycogen and muscle glycogen,the SOD activity and MDA level;the total inhibitory rate of oxidation of CSE in vitro was determined by linoleic acid-ferric thiocyanate method.Results:Compared with blank control group,the exhaustive swimming time of the mice in 50,100,and 200 mg·kg-1 CSE groups were significantly increased (P<0.01);the levels of BUN and LD of the mice in 100 and 200 mg·kg-1 CSE groups were significantly decreased (P<0.05 or P<0.01);the levels of liver glycogen and muscle glycogen of the mice in 100 and 200 mg·kg-1 groups were significantly increased(P<0.05 or P<0.01);whereas the SOD activities were significantly increased and the levels of MDA were decreased(P<0.05 or P<0.01).Compared with 100 mg·kg-1 CSE group,the levels of serum BUN and LD of the mice in 200 mg·kg-1 CSE group were decreased (P<0.01),and the levels of liver glycogen and muscle glycogen were increased(P<0.05).The total inhibitory rate of oxidation of 5 g·L-1CSE was 76.94%.Conclusion:CSE has an anti-fatigue effect and the mechanism may be related to anti-oxidation effect.
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Objective:To establish a method based on the iPLEX analysis of MassARRAY mass spectrometry platform to detect multiplex genetic mutations among Chinese lung cancer patients. Methods:We reviewed the related literature and data of lung cancer treatments. We also determined 99 mutation hot spots in 13 target genes, namely, EGFR, KRAS, ALK, FGFR1, FGFR2, FGFR3, PIK3CA, BRAF, PTEN, MET, ERBB2, AKT1, and STK11, which are closely related to the pathogenesis, drug resistance, and metastasis of lung cancer and are associated with relevant transduction pathways. A total of 297 primers comprising 99 paired forward and reverse amplification primers and 99 matched extension primers were designed by using Assay Design in accordance with the mutation label and format requirements of the MassARRAY platform. The detection method was established by analyzing eight cell lines and six lung cancer specimens;the proposed method was then validated through comparisons with a LungCarta kit. The sensitivity and specificity of the proposed method were evaluated by directly sequencing EGFR and KRAS genes in 100 lung cancer cases. Results:The proposed method could detect multiplex genetic mutations in the lung cancer cell lines, and this finding is consistent with that observed using previously reported methods. The proposed method could also detect such mutations in clinical lung cancer specimens;this result is also consistent with that observed by using the LungCarta kit. However, an FGFR2 mutation was detected only by using the proposed method. The measured sensitivity and specificity were 100%and 96.3%, respectively. Conclusion:The proposed MassARRAY technology-based method could detect multiplex genetic mutations among Chinese lung cancer patients. Indeed, the proposed method can be potentially applied to detect mutations in cancer cells.
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Objective To copy the mouse aging model with D-galactose,and to investigate the role of Schisandra total lignin (SCL)in the mouse brain tissue aging and its mechanism.Methods 50 mice were radomly divided into control group,model group (100 mg·kg-1 ·d-1),low dose (35 mg·kg-1 ·d-1)of SCL group (SCL-L), middle dose (70 mg· kg-1 · d-1 )of SCL group (SCL-M)and high dose (140 mg· kg-1 · d-1 )of SCL group (SCL-H)(n=10).D-galactose (100 mg·kg-1 ·d-1 )was injected into the mice hypodermically for 10 weeks to induce aging models in all the groups except control group,and 35,70,and 140 mg· kg-1 · d-1 SCL were administered for 10 weeks in SCL groups.The learning and memory abilities were measured by the Water Maze test.The expression levels of Bax,Bcl-2,ubiquitin (Ub),microtubule-associated protein light chain 3 (LC3)in the brain tissue of the mice in various groups were observed by Western blotting method. The LC3 protein expressions in mouse brain cortex and hippocampus were observed by immunohistochemistry.Results In learning and memory test,compared with control group,the swimming time of the mice in model group was increased (P<0.05),and the number of errors was increased (P<0.05);compared with model group,the swimming time in SCL-L,SCL-M and SCL-H groups was decreased (P<0.05)and the number of errors was also decreased (P<0.05). Compared with control group,the expression level of Bax was increased (P<0.05),the expression level of Bcl-2 was decreased (P<0.05),the expression levels of Ub and LC3-Ⅱ/LC3-Ⅰ proteins were increased (P<0.05)in model group;compared with model group,the expression level of Bax was decreased (P<0.05),the expression level of Bcl-2 was incerased (P<0.05),and the expression levels of Ub and LC3-Ⅱ/LC3-Ⅰ proteins were decreased (P<0.05)in SCL-L,SCL-M and SCL-H groups.In control group,the neuronal morphology was normal,and none of brown granules were visible in the cytoplasm of mouse brain cortex and hippocampus and the expression of LC3 protein was negative.In model group,the neurons were degeneration,and the number of LC3 protein positive cells in the cerebral cortex and hippocamptal tissue was increased (P<0.05).In SCL-L,SCL-M and SCL-H groups,the number of degenerative neurons was decreased,and the number of LC3 protein positive cells was decreased (P<0.05).Conclusion SCL can inhibit the D-galactose-induced brain tissue aging in the mice, and the mechanism is related to regulating autophagy and inhibiting apoptosis.
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ObjectiveTo investigate the fusion sequence complexity of EML4-ALK in non-small cell lung cancer (NSCLC) patients,and the potential mutation in tyrosine kinase ( TK ) domain of ALK gene.MethodsIn routine practice,a novel echinoderm microtubule-associated protein-like4 and anaplastic lymphoma kinase (EML4-ALK) V3c variant was detected by rapid amplification of cDNA ends-polymerase chain reaction ( RACE-PCR )-sequencing technology in a patient with NSCLC.The further consecutive 39 cases( total of 40 cases)were screened by use of reverse transcription (RT)-PCR for EML4-ALK fusion.Positive PCR products were purified and cloned into T vectors,transformed into DH5a germ cells and colony picked up and sequenced for sequence complexity analysis.Tyrosine kinase domain of ALK was amplified by RT-PCR and sequenced.ResultsThree out of 40 cases had EML4-ALK fusion.One case had six novel variants of EML4-ALK co-existing,termed as V3c ( 64.6％ ),V3d ( 25.0％ ),V3e ( 2.1％ ),V3f (4.2％ ),V3g(2.1％ )and V3h(2.1％ ) variants,whereas without common V3a and V3b variants.In other two positive cases,one was V1 variant,another was concurrent V2,V3a and V3b variants.No mutations were detected in the TK domain of EML4-ALK in any case.ConclusionsSeveral EML-ALK variants could co-exist in a given lung cancer tissue,which suggest that the diversity and sequence complexity of EML4-ALK fusion are exist.Attentions should be paid to screen all the variants in clinic to improve the pick-up rate.
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ObjectiveTo investigate the D8/17 antigen expression of patients with rheumatic heart disease (RHD) in Guangdong province and study the antigen's characteristics.Methods The level of D8/17 antigen expression on B lymphocytes was determined with flow cytometry assay in 96 RHD patients and 83 unaffected controls.The percentage of B-cells expressing the D8/17 antigen having more than 10％ was considered to be positive. D8/17 antigen was extracted by immunopreeipitation,and the antigen characteristics was analyzed by tandem mass spectrometry. Results The mean percentage of B-cells expressing the D8/17 antigen was (85.36 ± 15.15)％ in the RHD patients and (82.89 ±4.55)％ in the controls,with no significant difference between the two groups ( P =0.436).Moreover,the positive rate of the D8/17 cxprcssion was 100％ in either the RHD patients or the controls.The molecular weight of D8/17 antigen was found to be 40 000-67 000,and the purified protein was most likely to match moesin or β-actin.ConclusionsB-cell antigen D8/17 is not associated with RHD in Guangdong province of China.Moesin or β-actin is the most likely protein to match D8/17 antigen.
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Objective To investigate the protective effect of Schizandrae Lignanoid(SCL)on the apoptosis of PC12 cells induced by H2O2 and its relative mechanisms.Methods PC12 cells were divided into four groups: control group,model group,high dose SCL(SCL1) group,and low dose SCL(SCL2) group.Apoptosis of PC12 cells was induced by H2O2.The cell activity was determined by MTT,the cell apoptotic rate was determined by Annexin V-FITC/PI and ??m was detected by flow cytometry.The expressions of bcl-2 and bax were detected by immunohistochemistry.Results Compared with model group,SCL increased the survival rate of PC12 cells(P
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Objective To investigate the protective effect of Schizandrae Lignanoid (SCL) on myocardial ischemia reperfusion injury in hyperlipidemic rats and its mechanisms and provide theoretical basis and experiment foundation for treatment of myocardial ischemia complicated with hyperlipidemic disease with SCL in the future. Methods After hyperlipidemic rat models were set up by oral administration of high lipid emulsion,the rats were divided into seven groups: control group,sham group,model group,SCL 60,20,5 mg?kg-1 groups and 200 mg?kg-1 CDT group.After reperfusion,the blood samples taken from the ventricles were assayed for blood lipid;MPO activities of ischemic myocardium were measured;Infarct area of left ventricles was measured by Evens blue-TTC staining,and myocardial pathological changes were also observed.Results Compared with model group, the myocardial infarct area/ventricle area ratios and MPO activities in SCL 60,20,5 mg?kg-1 groups decreased (P
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Objective To explore the epidemic conditions of rheumatic fever (RF) with a simple method at some regions.Methods Antistreptococcal DNase B test microtiter method was adopted to detect human sera antibody in natural populations who are responsible to RF.All 216 schoolchildren aged 10～12 years old were chose by the random cluster sampling.The schoolchildren′s serum antibody against DNase B in each season from september 1988 to August 1989 was detected,and compared with antistreptolysin “O” (ASO);furthermore,5 different regional schoolchildren′s serum antibody to DNase B was tested, and 100 schoolchildren aged 10～13 years old were chose in every region.Meanwhile,the RF incidence of 5～18 year schoolchildren was investigated for 4 years,and approximately 364 915 person times were inquired.All the results of the anti DNase B and ASO were compared with the schoolchildren′s RF incidence.Results ① The levels of the anti DNase B were higher during fall,winter and spring seasons,and lower in summer,which were coincided with the RF incidence,and linear relation was very good ( r =0 913, P
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At the concentrations of 10-8mol/L, 5.5?l0-8mol/L, 10-7mol/L morphine produced a dose-dependent increase of beating rate in cultured rat heart cells with 9%,32%and 81% respectively. The increase was antagonized by opiate antagonist naloxone ( 10-5mol/L ) , but not affected by propranolol ( 10-5mol/L ) which could cancel the positive chronotropy of isoprenaline ( 10-9mol/L ) on the beating cells. The results support the concept that morphine may be a direct effect on myocadium, perhaps through opiate receptors.