3.Investigation on influencing factors of 5-HMF content in Schisandra.
Qing XU ; Ying-Hua LI ; Xiu-Yang LÜ
Journal of Zhejiang University. Science. B 2007;8(6):439-445
In order to investigate the influencing factors of 5-hydroxymethyl-2-furaldehyde (5-HMF) content in Schisandra, confirm the theory of 5-HMF deriving mainly from Schisandra processing course, and give some suggestions about the Schisandra processing method, the 5-HMF contents in decoctions of Schisandra under different heating temperature, decocting time, soaking time, processing methods and treatment with different solvents before decocting the Schisandra were measured by RP-HPLC method. The results showed that there is great difference of 5-HMF level in decoctions from differently processed Schisandra and unprocessed Schisandra; decocting time of 60 min has some effects on 5-HMF level in decoctions and there is certain quantity 5-HMF in processed Schisandra itself and very little 5-HMF in unprocessed Schisandra. Heating time, heating temperature and treating solvents all have effect on 5-HMF level in decoction of Schisandra. 5-HMF in Schisandra was mainly from processing course. Both long heating time and high heating temperature can increase 5-HMF level in Schisandra. The production of 5-HMF in Schisandra may have some relationships with some polar components, which can dissolve in water, ethanol and acetone, especially in ethanol. To control processing temperature, processing time and treatment with some solvent is very important for controlling 5-HMF level in Schisandra.
Chromatography, High Pressure Liquid
;
Furaldehyde
;
analogs & derivatives
;
analysis
;
Plant Extracts
;
chemistry
;
Schisandra
;
chemistry
;
Temperature
;
Time Factors
4.Clinical observation on treatment of cervicogenic headache with turtle-probing needling at Tianzhu (BL 10).
Chinese Acupuncture & Moxibustion 2006;26(11):796-798
OBJECTIVETo observe therapeutic effect of turtle-probing needling at Tianzhu (BL 10) on cervicogenic headache.
METHODSSeventy cases were randomly divided into a treatment group (n=36) and a control group (n= 34). The treatment group were treated with turtle-probing needling at Tianzhu (BL 10) on the affected side, first perpendicularly needling the point and then penetration-needling homolateral Fengchi (GB 20), Fengfu (GV 16), below cervical Jiaji points (EX-B2), and occipital tuberosity in order. And the control group were treated with acupuncture at Fengchi (GB 20),Tianzhu (BL 10), Shuaigu (GB 8), Touwei (ST 8) and corresponding cervical Jiaji points (EX-B2), Houxi (SI 3), Ashi points at the affected side with routine needling manipulation. Treatment was given once each day, 6 sessions constituting one course.
RESULTSThere was no significant difference between the two groups in therapeutic effect (P > 0.05), but the cured and markedly effective rates for analgesic effect in the observation group at the first three sessions were higher than those in the control group (P < 0.05).
CONCLUSIONThe turtle-probing needling method and the routine needling have a similar therapeutic effect, but the turtle-probing needling method more rapidly produces the analgesic effect.
Acupuncture Points ; Acupuncture Therapy ; methods ; Adolescent ; Adult ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Post-Traumatic Headache ; therapy
5.Efficacy comparison of different forms of dog-days acupoint application for prevention and treatment of chronic bronchitis
Yang LIU ; Lin-Hua CUI ; Qing LI ; Xiao XING ; Cong-Hui QI ; Shao-Jin WANG ; Ying LÜ
Journal of Acupuncture and Tuina Science 2019;17(3):168-173
Objective: To analyze the clinical effect of traditionaldog-days acupoint application and Magic Acupuncture Patch (Manji) in the prevention and treatment of chronic bronchitis (CB) in remission stage in the past five years, and explore the principle of action and effective stimulation, to provide the evidence for treating CB by acupoint application. Methods: A retrospective study was conducted on 405 patients with CB who met the inclusion criteria. All patients were treated with dog-days acupoint application or Magic Acupuncture Patch between 2013 and 2017. The clinical data of 405 patients were statistically analyzed to compare the prevention and treatment effects of dog-days acupoint application and Magic Acupuncture Patch, and different degrees of stimulation of dog-days acupoint application. Results: Among the dog-days acupoint application groups, the total effective rate was 63.6% in the light stimulation group, 93.1% in the moderate stimulation group, and 94.8% in the strong stimulation group. The differences in the total effective rate between the light stimulation group and the moderate stimulation group, as well as the strong stimulation group, were statistically significant (both P<0.05). There was no significant difference in the total effective rate between the moderate stimulation group and the strong stimulation group (P>0.05). The total effective rate was 83.9% in the dog-days acupoint application group, versus 45.4% in the Magic Acupuncture Patch group, and there was a statistically significant difference between the two groups (P<0.05). Conclusion: The efficacy of dog-days acupoint application in the prevention and treatment of CB is better than that of Magic Acupuncture Patch; the degree of stimulation is the basis for the effect of dog-days acupoint application on prevention and treatment of CB, and the moderate and strong stimulations are more appropriate.
6.Investigation of RHD 1227A allele in five pedigrees in Zhejiang Han population.
An-xin CHEN ; Hao LÜ ; Feng-juan XU ; Li-ying ZHANG ; Ying-hua NI ; Hong-hui YE ; Ying ZHAO
Chinese Journal of Medical Genetics 2006;23(5):536-539
OBJECTIVETo analysis the genetic mode of Rh DEL phenotype and RHD 1227A allele in Zhejiang Han population through family investigations.
METHODSRh DEL phenotypes were identified by a serologic adsorption-elution method. Two polymerase chain reaction-sequence specific prime (PCR-SSP) methods which detectED RHD 1227A allele and Rhesus hybrid box, respectively, and a nucleotide sequencing method focused on the exon 9 of RHD 1227A allele were employed to determine the zygosity of RHD allele.
RESULTSAll five probands with Rh DEL phenotype harbored a RHD 1227A allele and had a RHD allele deletion, and they were RHD 1227A/RHd heterozygote. One of the parent members was found to contain a RHD 1227A allele and a normal RHD allele in pedigree 1, 2 and 3, respectively. Thus, they were RHD 1227A/RHD heterozygotes and presented normal D positive phenotype. The son of proband No 1. inherited the RHD 1227A allele and presented a normal D positive phenotype due to a RHD 1227A/RHD heterozygote; The offsprings of proband No. 2, No. 4, and No. 5 did not inherit RHD 1227A allele and presented a normal D positive phenotype.
CONCLUSIONRHD 1227A allele is an important genetic marker of Rh DEL phenotype; RHD 1227A is recessive to normal RHD allele and dominant to RHd allele; RHD 1227A allele is an ancestral, but not a spontaneously mutated allele.
Alleles ; China ; Female ; Genotype ; Humans ; Male ; Pedigree ; Phenotype ; Polymerase Chain Reaction ; methods ; Rh-Hr Blood-Group System ; genetics
7.Effects of transforming growth factor beta 1 on the growth of rhabdomyosarcoma cell line RD.
Lü YE ; Hong-ying ZHANG ; Hua WANG ; Guang-hua YANG ; Hong BU ; Li ZHANG ; Shou-li WANG
Chinese Medical Journal 2005;118(8):678-686
BACKGROUNDTransforming growth factor-beta (TGF-beta) can inhibit the growth of most epithelial and endothelial cells. The growth regulative role of TGF-beta on soft tissue sarcoma was seldom reported. Here we examined TGF-beta1 effects on the growth of human rhabdomyosarcoma cell RD and searched the relative molecular mechanism.
METHODSThe viability of RD was examined by [(3)H]-thymidine incorporation and [3-(4,5-dimethylthiazol-z-yl)-2,5-diphenyl tetrazolium bromide] (MTT) assay. RD cell cycle was analysed by flow cytometry. The protein and mRNA of cell cycle regulative factors in RD were detected by Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. The kinase activity of cdk2 or cdk4 was examined by immunoprecipitation and kinase assay. Immunofluorescent staining was used to detect the location of cell cycle regulative factors in RD by laser scanning confocal microscope.
RESULTSTGF-beta1 inhibits RD proliferation by G1-arrest in cell cycle progression. TGF-beta1 can prominently up-regulate P27 of RD, then augment P27 to bind cyclinE-cdk2 complexes, which effectively suppress cdk2 kinase activity. P21 increased and c-myc decreased in RD due to TGF-beta1. Both P15 and cdk4 have not been involved in the growth inhibitory event. TGF-beta1 treatment induced P27 to congregate around nucleus. P21 pervaded from nucleus to both nucleus and cytoplasm by TGF-beta1 treatment.
CONCLUSIONTGF-beta1 inhibits the proliferation of human rhabdomyosarcoma cell line RD and induces RD G1-arrest. This course is accomplished by TGF-beta1 up-regulating P27 to suppress cdk2 kinase activity. The induction of P21 and down-regulation of C-myc might participate in the growth-arrest event.
Cell Cycle Proteins ; analysis ; genetics ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p27 ; G1 Phase ; drug effects ; Genes, myc ; Humans ; RNA, Messenger ; analysis ; Rhabdomyosarcoma ; pathology ; Transforming Growth Factor beta ; pharmacology ; Transforming Growth Factor beta1 ; Tumor Suppressor Proteins ; analysis ; genetics
8.Regulation of growth inhibition by transforming growth factor beta1 in rhabdomyosarcoma RD cell line.
Lü YE ; Hong-Ying ZHANG ; Hong BU ; Guang-Hua YANG ; Shou-Li WANG ; Hua WANG
Chinese Journal of Pathology 2004;33(6):541-545
OBJECTIVETo study the regulatory effect of TGF-beta1 on growth of rhabdomyosarcoma RD cell line.
METHODSAfter various durations of TGF-beta1 treatment, the viability of RD cell line was examined by growth rate measurement, MTT assay and (3)H-thymidine incorporation. The cell cycle was analyzed by flow cytometry. Immunofluorescent staining was used to localize p15, p21 and p27 in RD cell line under laser scanning confocal microscope. The protein and mRNA of p15, p21 and p27 in RD cell line were detected by western blot and reverse transcriptase-polymerase chain reaction respectively.
RESULTSThe viability of RD cell line treated with TGF-beta1 was obviously decreased. RD cell line was arrested in G(1) phase by TGF-beta1. There was increased expression of p21 and p27 in RD cell line with TGF-beta1 treatment at protein and mRNA levels. The expression of p21 in RD cell line was seen in both nucleus and cytoplasm after 24 hours of TGF-beta1 treatment. The expression of p15 showed no obvious changes upon TGF-beta1 treatment.
CONCLUSIONSTGF-beta1 inhibits growth of RD cell line and induces G(1)-arrest. It up-regulates protein and mRNA of p21 and p27 and shows no obvious influence on p15 expression. The growth arrest of RD cell line may result from the up-regulation of p21 and p27 by TGF-beta1.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p15 ; biosynthesis ; genetics ; Cyclin-Dependent Kinase Inhibitor p21 ; biosynthesis ; genetics ; Cyclin-Dependent Kinase Inhibitor p27 ; biosynthesis ; genetics ; G1 Phase ; drug effects ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; Rhabdomyosarcoma ; pathology ; Transforming Growth Factor beta1 ; pharmacology
9.Smad4 and ERK2 stimulated by transforming growth factor beta1 in rhabdomyosarcoma.
Hua GUO ; Hong-ying ZHANG ; Shou-li WANG ; Lü YE ; Guang-hua YANG ; Hong BU
Chinese Medical Journal 2007;120(6):515-521
BACKGROUNDTransforming growth factor beta (TGF-beta) plays an essential role in the regulation of normal physiologic processes of cells. TGF-beta has been shown to regulate several mitogen-activated protein kinases (MAPK) pathways in several epithelial cells. However, the effects of TGF-beta on soft tissue sarcoma are seldom reported. Our previous studies suggested that there should be some other signal transduction pathways besides Smads, which are important to regulate the growth of human embryonal rhabdomyosarcoma (RMS) cells. In the present study, we examined the expression and functional relations of extracellular signal-regulated kinase 2 (ERK2) and Smad4 in human RMS tissue and a RMS cell line, RD.
METHODSRD cells and normal human primary skeletal myoblasts (Mb) were treated with TGF-beta1 to establish the expression profile of ERK2 at the mRNA and protein levels detected by RT-PCR and immunofluorescence. Immunohistochemistry was used to detect the expression of ERK2 and Smad4 in 50 tissue specimens of human RMS and 23 specimens of normal skeletal muscles. Follow-up of specimens was performed 6 months to 70 months later.
RESULTSRD cells and human RMS tissues showed the higher expression of ERK2 and Smad4 than the normal control, either the protein level or the mRNA level. And, exogenous TGF-beta1 stimulation can lead to higher expression of ERK2 and its nuclear translocation, so TGF-beta1 can also activated MAPK (ERK2) pathway, resulting in a sustained activation of ERK2 for at least 2 hours. Immunohistochemistry analysis, however, showed that there was no correlation between ERK2 and Smad4 protein. The overexpression of ERK2 and Smad4 had no indicative effects on histological subtypes, histological grading, gender, age, and prognosis.
CONCLUSIONSIn RMS, signaling of TGF-beta1 from cell surface to nucleus can also be directed through the MAPK (ERK2) pathway besides the TGF-beta1/Smads pathway. The activation of ERK2 by TGF-beta1 may be Smad4 independent. Moreover, there may be some other tanglesome relationships between the TGF-beta1/Smads pathway and the MAPK pathway which takes part in the development, invasion and metastasis of tumor cells.
Cells, Cultured ; Humans ; Mitogen-Activated Protein Kinase 1 ; physiology ; Muscle, Skeletal ; metabolism ; RNA, Messenger ; analysis ; Rhabdomyosarcoma ; metabolism ; Signal Transduction ; Smad4 Protein ; physiology ; Transforming Growth Factor beta1 ; pharmacology
10.Effects of transforming growth factor-beta/Smad signaling on the growth and apoptosis of human rhabdomyosarcoma cell line RD.
Lü YE ; Hong-ying ZHANG ; Guang-hua YANG ; Hong BU ; Hua GUO
Chinese Journal of Pathology 2005;34(7):407-412
OBJECTIVETo study the effects of TGF-beta/Smad signaling on the growth and apoptosis of human rhabdomyosarcoma cell line RD.
METHODSBiosynthesized short hairpin RNA (shRNA) was transfected into RD cells by cation liposome vector to suppress Smad4 expression. The mRNA and protein expression of Smad4 in RD after shRNA-transfection were examined by RT-PCR and Western blot respectively. Immunofluorescent staining was used to detect the location of Smad2/3 in RD by laser scanning confocal microscopy. The viability of RD cells was examined by MTT method and (3)H-thymidine incorporation assay. The apoptosis of RD was examined by flow cytometry analysis and fluorescent staining.
RESULTSThe expression of mRNA and protein of Smad4 in RD were effectively suppressed by shRNA interference. Such suppression effectively interrupted the endogenous TGF-beta/Smad signaling and consequently blocked the translocation of Smad2/3. The interruption of endogenous TGF-beta/Smad signaling not only inhibited the growth of RD but also induced apoptosis of RD. Exogenous TGF-beta1 inhibited the growth of RD but did not influence the apoptosis of RD.
CONCLUSIONshRNA interference can effectively interrupt the TGF-beta/Smad signaling by suppressing the expression of Smad4. TGF-beta/Smad signaling subtly regulates the growth and apoptosis of RD.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Cell Proliferation ; drug effects ; Cytoplasm ; metabolism ; Humans ; Protein Transport ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; pharmacology ; Rhabdomyosarcoma ; genetics ; metabolism ; pathology ; Signal Transduction ; Smad2 Protein ; metabolism ; Smad3 Protein ; metabolism ; Smad4 Protein ; biosynthesis ; genetics ; Transfection ; Transforming Growth Factor beta1 ; pharmacology