Objective To investigate the effect of sodium butyrate on the growth suppression and cellular differentiation of cultured SHG 44 glioma cell line Methods Human glioma SHG 44 cells were treated with 2 mmol/L sodium butyrate The inhibition of cell growth was assessed by MTT spectrophotometric analysis and AgNORs analysis The cellular differentiation was determined by flow cytometry, electron microscopy, immunohistochemical method and agglutination test Results Treatment of SHG 44 cells with sodium butyrate resulted in significant inhibition of cell growth and G1 S transition in time dependent and dose dependent manners Sodium butyrate could induce differentiation of SHG 44 cells with regularized nuclei and increased heterochromatins The mitochondria and rough endoplasmic reticulum increased in density in normal state Furthermore, SHG 44 cells expressed more GFAP but less vimentin, and the agglutination degree decreased obviously Conclusion Sodium butyrate may suppress glioma cells growth and induce differentiation of human glioma cells

In order to increase the yield and quality of the medicinal plant and enhance the competitive power of industry of medicinal plant in our country, this paper analyzed the status, problem and countermeasure of the tissue culture of medicinal plant on large scale. Although the biotechnology is one of the most efficient and promising means in production of medicinal plant, it still has problems such as stability of the material, safety of the transgenic medicinal plant and optimization of cultured condition. Establishing perfect evaluation system according to the characteristic of the medicinal plant is the key measures to assure the sustainable development of the tissue culture of medicinal plant on large scale.
Drug Industry ; methods ; standards ; Medicine, Chinese Traditional ; methods ; standards ; Plants, Genetically Modified ; Plants, Medicinal ; genetics ; growth & development ; Quality Control ; Tissue Culture Techniques ; methods ; standards

BACKGROUND: Preactivation of peripheral blood mononuclear cells (PBMCS) is one of the most important eerly events and facilitating factor for the formation of atherosclerosis. Tanshinone is a lipolytic component extracted from traditional Chinese medicine of denshen, it has definite anti-atherosclerotic effect.OBJECTTVE: To analyze whether PBMCS preactivation existed at early essential hypertension, and investigate the effects of tanshinone on inhibiting the PBMCS activation cultured in vitro by detecting the adhesion and excretory activities of PBMCS.DESTGN: A case-controlled analysis.SETTING: Department of Emergency and Research Room of Traditional Chinese Medicine, Tongji Hospital affiliated to Tongji Medical College, Huazhong University of Science and Technology.PARTTCTPANTS: Thirty patients with untreated essential hypertension or with withdrawal from antihypertensives for at least 2 weeks were selected from the Department of Cardiology, Tongji Hospital affiliated to Tongji Medical College,Huazhong University of Science and Technology from January 2003 to October 2004, including 16 males and 14 females, aged (44.6±7.4) years, body mass index of (26.2±4.5) kg/m2, average disease course of (38.5±16.9) months.Informed contents were obtained from all the subjects. Their hypertension was grade Ⅰ-Ⅱ according to the diagnostic standards for hypertension by WHO/ISH in 1999. Secondary hypertension, organic heart disease, hyperglyceridemia,diabetes mellitus, liver and kidney dysfunction, heart, brain, kidney, vessel and other target damaged induced by infection and other clinical conditions and hypertension were excluded by history, physical examination and assistant examination.Another 30 healthy physical examinees with normal blood pressure were enrolled as the normal control group. Human umbilical vein endothelial cells (Species Reserving Center of Wuhan University); Tanshinone injection (Yaan Sanjiu Pharmaceutical, Co., Ltd., batch number: 020724);METHODS: ① Venous blood samples (4.0-5.0 mL) were drawn from all the subjects, and mononuclear cells were separated by means of Ficoll-Hypaque density gradient centrifugation and plastic adhesion, then incubated at 37 ℃ for 40-60 minutes, and the adherent cells were the PBMCS. These cells (viability ＞ 95%, Trypan blue staining) had the characteristics of mononuclear cells (Wright staining). The newly separated adherent PBMCS were resuspended, and then inoculated to the 24-well plate (4×107 L-1). There were 3 wells for each sample: the first was for basic excretion, the second for angiotensin Ⅱ stimulation, and the third for tanshinone pretreatment. The PBMCS were co-incubated with tanshinone for 30 minutes before angiotensin Ⅱ stimulation. The terminal concentration was 1×10-8 mol/L and 1×10-8 g/L for angiotensin Ⅱ andtanshinone respectively, and that of PBMCS was 2×107 L-1. The cells were cultured in the incubator (CO2 of 0.05 in volume fraction) at 37 ℃ for 24 hours, then the supernatant and cell ingredients were collected respectively. ② The PBMCS suspension was preparedl, and the cellular density was adjusted to 2.5×109 L-1. The human umbilical vein endothelial cells were cultured on the 24-well plate with M199 medium containing fetal bovine serum (0.1 in volume fraction), and spread to monolayer after the cells entered the logarithm phase. Each well was added with PBMCS suspension (100 μL), incubated at 37 ℃ for 2 and 4 hours respectively. The unadherent cells were removed, and the adherent ones were counted after fixed with 20 g/L glutaral, 40 visual sights were counted for each well under high power microscope, and the average value was used. ③ The double-antibody sandwich enzyme-linked immunoabsorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) were used to determine the concentrations of tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β) and interleukin-6 (IL-6) in supernatant of PBMCS, and the expressions of their mRNA.MATN OUTCOME MEASURES: ① Changes of PBMCS adhesion activity; ② Concentrations of cytokines and their mRNA expressions in supernatant of PBMCS.RESULTS: ① At 2 and 4 hours of inoculation, the numbers of PBMCS adhered to endothelial cells under basic conditions were similar between the hypertension group and normal control group (t =1.153-1.577, P ＞ 0.05); After angiotensin Ⅱ stimulation, the adherent cells were obviously more in the hypertension group than in the normal control group (t =3.842-4.536, P ＜ 0.01); The numbers of the adherent cells were decreased to the same levels after tanshinone pretreatment (t =0.855-1.702, P ＞ 0.05). ②Under basic state, the concentrations of TNF-α, IL-1β and IL-6 in supernatant were all lower in both groups (t =0.981-1.829, P ＞ 0.05); The concentrations of the cytokines after angiotensin Ⅱ stimulation were obviously higher in the hypertension group than in the normal control group (t = 2.442, 5.075, P ＜ 0.01,0.01, 0.05); The concentrations of the cytokines after tanshinone pretreatment were all decreased to different extent, and there were no significant differences (t =1.227-1.940, P ＞ 0.05). Similar changes were observed in the mRNA expressions of the cytokines in PBMCS in the two groups.CONCLUSTON: ① The number of PBMCS adhered to endothelial cells, the concentrations and mRNA expressions of the cytokines under basic state in patients with essential hypertension were at the levels of normal subjects, and they were significantly increased after angiotensin Ⅱ stimulation, suggesting that the PBMCS were at preactivation at early essential hypertension. ② Tanshinone could decrease the adhesion and excretory activities of PBMCS in patients with essential hypertension to the normal levels, it is proved that tanshinone can inhibit the further activation of the preactivated PBMCS, and can prevent the occurrence of atherosclerosis to some extent.

Endocardial Cushion Defects ; complications ; Female ; Humans ; Infant, Newborn ; Lung ; abnormalities ; Lung Diseases ; complications ; congenital

BACKGROUND: +Gz-induced acute dysencephalia and its protection is one of the significant topics in Aero-medical researches. Its pathological mechanism, however, is still unclear and protective measures should be developed further. OBJECTIVE: To observe the expression of inducible nitricoxide synthase (iNOS) in brain tissue after +Gz exposure and to analyze the protective effects of danshen and basic fibroblast growth factor (bFGF) on repeated +Gz exposure-induced brain injury. DESIGN: Randomized controlled animal study. SETTING: Researching Center of Molecular Biology, Air-force General Hospital of Chinese PLA.MATERIALS: The experiment was carried out at the Researching Center of Molecular Biology, Air-force General Hospital of Chinese PLA from April to August 2000. A total of 20 healthy SD rats of clean grade were divided into 5 groups according to randomly digital table, including control group, +Gz exposure group, bFGF group, danshen group and saline group with 4 in each group.METHODS: All rats were fixed on rotatory arm of centrifugal apparatus,and their heads were towards core of the apparatus. Except the rats in control group, the value of +Gz exposure was +14 Gz, and the growth rate was 1.5 G/s. The exposure at peak value lasted for 45 s. +Gz exposure was done for three times, and the interval was 30 minutes. Rats in the control group were also treated with the same +Gz exposing procedure, but the G value was +1 Gz. Rats in bFGF group and danshen group were intraperitoneally injected with 100 μg/kg of bFGF and/or 15 g/kg of danshen solution, respectively, at 30 minutes before centrifugation and immediateness after centrifugation; moreover, rats in saline group were injected with the same volume of saline. Six hours after exposure, rats were cut off their heads to obtain the brains which were maintained in liquid nitrogen for RNA extraction. The expression of iNOS mRNA in brain tissues of the rats in each group was detected with semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and calculated on the basis of ratio between iNOS and glyceraldehyde-3-phosphate dehydrognase.MAIN OUTCOME MEASURES:Expressed level of iNOS mRNA in brain tissue of rats.RESULTS: Expression of iNOS mRNA in brain tissue was higher in repeated +Gz exposure group than that in control group (0.452 ±0.014,0.065±0.008, P ＜ 0.01); however, that was lower in bFGF group and dan-shen group than that in +Gz exposure group (0.196±0.010, 0.183±0.011,0.452±0.014, P ＜ 0.01).CONCLUSION: Repeated +Gz exposure can increase the expression of iNOS mRNA, this plays an important role in cerebral injury induced by repeated +Gz exposure. Moreover, bFGF and danshen have protective effects on cerebral injury induced by +Gz exposure.

Objective To explore the relation between the level of oxygen pressure and lactate in arterial blood and prognosis of cardiopulmonary resuscitation (CPR).Methods 36 patients with sudden cardiac arrest received CPR were selected.Patients were divided into 3 groups.The 21 patients who 30 minutes of CPR didn't regain heart beats were group A,those 10 who kept blood circulation for over 2 hours but finally died were group B,and the remaining 5 who survived were group C.Control group was made up of 20 people with no disease.In CPR immediately (0h),1h,3h,6h,12h were measured in blood specimens from arterial partial pressure of oxygen and lactate levels were compared among groups of patients dynamic changes of the index.Results The 3 groups A,B,C significantly reduced blood oxygen pressure,blood lactate increased significantly the CPR in the early,and compared with the control group there was significant difference( P ＜ 0.01 ).The group B CPR 3h,6h,and continue until no significant changes in 12h;The group C CPR 3h,6h began to increase blood oxygen pressure and blood lactate levels begin to decline,blood oxygen pressure increased significantly,lower blood lactate to a lower level after 12h.Taken with marked changes in B group,the difference was significant ( P ＜ 0.05 ).Conclusion Arterial partial pressure of oxygen and blood lactate levels could be used as indicators of prognosis of patients with CPR.Sustained increase in arterial oxygen partial pressure showed a good prognosis,lactate levels continued to rise that a poor prognosis.

BACKGROUND: Aldosterone is an important pathogenic factor of left ventricular hypertrophy. There has been evidence that the extract of red sage root (a Chinese herb) can intervene the synthesis of aldosterone.OBJECTIVE: To investigate the effects of tanshinone Ⅱ A on expression of genes related to aldosterone synthesis in myocardium. DESIGN:A randomized and controlled trial. SETTING :Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology. MATERIALS:The experiment was performed at the laboratory of Emergency Department, Tongji Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology from November 2002 to March 2004. Totally 20 male 12-week-old rats with spontaneous hypertension were randomly divided as hypertension group and tanshinone Ⅱ A group. METHODS:Rats in each group were injected tanshinone Ⅱ A or distilled water in the same volume respectively through caudal vein. After 12-week administration,the rats were put to death by decapitation, and the samples of myocardium were prepared. The expressions of CYP11B1mRNA and CYP11B2 mRNA, or myocardial genes related to aldosterone synthesis were examined with reverse transcriptase polymerase chain reaction (RTPCR) and referring to the amplification primers of glyceraldehydes 3-phosphate dehydrogenase (GAPDH) gene. MAIN OUTCOME MEASURES:The levels of CYP11B1 and CYP11B2 mRNA related to aldosterone synthesis in myocardium.RESULTS :Totally 20 rats were involved in the trial and all entered in the final result analysis without any loss of value. ① Quantitative analysis of expression of myocardial CYP11B1 and CYP11B2 genes in rats of two groups: Taking 100bp Plus Ladder as Marker,clear amplified strands could be seen at 440bp,461bp and 336bp sites,DNA sequencing proved they were the encoding gene segments of CYP11B1,YP11B2 and GAPDH. ②Qualitative analysis of expression of myocardial CYP11B1 and CYP11B2 genes in rats of two groups:The expressive levels of CYP11B1 and CYP1 1B2 mRNA contents in tanshinone Ⅱ A group were significantly lower than those in hypertension group (0.924±0.121 vs 1.343±0.132,P ＜ 0.05;1.017±0.119 vs 1.675±0.126,P 0.01). CONCLUSION:Tanshinone Ⅱ A could directly down-regulate the expression of CYP1 1B1 and CYP1 1B2 mRNA, the genes related to aldosterone syn thesis, so that it inhibited the biological synthesis of aldosterone in myocardinm, thus playing a role to resist hypertensive left ventricular hypertrophy.

BACKGROUND:Aldosterone is main pathogenic factor of left ventricular hypertrophy, for which, to inhibit its biosynthesis effectively may prevent and treat hypertension induced by left ventricular hypertrophy.OBJECTIVE: To investigate the effects of Tanshinone Ⅱ A on myocardiac aldosterone synthesis and expression of its relevant genic CYP11B1and CYP1 1B2 in spontaneously hypertensive rats(SHR) and explore the possi ble nechanism of Tanshinone Ⅱ A on inhibiting hypertension induced by left ventricular hypertrophy.DESIGN:SHR were employed as the objects in the experiment and WKY rats with normal blood pressure (WKY rats) were employed in the controlgroup. Complete group division and randomized control experiment was designed. Analysis of variance was used for the means comparison among groups.SETTING :Department of Emergency and Institute of Integrated Traditional Chinese and Western Medicine in Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS :The experiment was performed in the laboratory of Department of Emergency in Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from January 2003 to February 2004, in which, 10 male WKY rats with normal blood pressure of 12-week-old were employed in the control group and 20 male SHR of 12-week-old were randomized into two groups, named hypertension group and Tanshinone ⅡA group, 10 rats each group.METHODS:In Tanshinone Ⅱ A group, Tanshinone Ⅱ A 1.5 mg/kg was injected abdominally everyday. The distilled water of same volume was injected abdominally in hypertension group and the control group. Twelve weeks after experiment, myocardial specimens were collected after rats sacrificed. Radioimmunoassay was used to determine the contents of aldosteron and angiotensin Ⅱ in myocardial tissue and reverse transcriptase polymerase chain reaction (RT-PCR) were used to measure mRNA level of CYP11B1 and CYP11B2 genes relevant to aldosterone synthesis.MAIN OUTCOME MEASURES: Contents of aldosterone and angiotensin Ⅱ in myocardial tissue and mRNA expression of CYP11B1 and CYP1 1B2 genes.RESULTS:All of 20 SHR and 10 WKY rats entered result analysis. ① Myocardial aldosterone content of rats in 12 weeks after experiment: that in hypertension group and Tanshinone Ⅱ A group was higher remarkably than that in the control group [(0.056±0.014), (0.031±0.010), (0.018±0.009) ng/g,P ＜ 0.01, 0.05]; that in Tanshione Ⅱ A group was lower remarkably than that in the hypertension group (P ＜ 0.05). ② Myocardial angiotension Ⅱ content of rats in 12 weeks after experiment: that in the hypertension group and Tanshinone ⅡA group was higher remarkably than that in the control group [(0.093±0.016), (0.088±0.024), (0.043±0.012) ng/L, P ＜ 0.01]. ③ Expression of CYP11B1 gene in myocardial tissue of rats in 12 weeks after experiment: that in the hypertension group and Tanshinone Ⅱ A group was higher remarkably than that in the control group (2.774±0.138, 2.533 ±0.127, 1.973±0.102, P ＜ 0.05). ④Expression of CYP11B2 gene in myocardial tissue of rats in 12 weeks after experiment; that in the hypertension group and Tanshinone ⅡA group was higher remarkably than that in the control group (1.573±0.106, 1.024±0.113, 0.786±0.121, P ＜ 0.01,0.05); that in the Tanshinone ⅡA group was lower remarkably than that in the hypertension group (P ＜ 0.05). ⑤The electrophoresis band positions of myocardial CYP11B1 and CYP11B2 as well as RT-PCR products of glyceraldehydes-3-phosphate dehydrogenase (GAPDH), the consulting gene,were in conformity with the theoretic values.CONCLUSION:The inhibition of Tanshinone Ⅱ A on hypertension induced by left ventricular hypertrophy is probably contributed to its downregulating effect on myocardiac CYP11B2 gene expression relevant to aldosterone synthesis and to its reducing action on local biosynthesis of aldosterone in heart.

BACKGROUND: As a reliable marker of inflammation, intercellular adhesion molecule-1 (ICAM-1) plays an important role in atherogenesis. Recently, it is assumed in researches of recent years that chronic inflammation mediated by ICAM-1 is involved in hypertensive left ventricular hypertrophy probably, but there are still some objections reported. Tanshinone ⅡA is a kind of liposoluble extract from danshen(Radix Salviae Mitiorrhizae) . It is verified in animal experiment that it can inhibit hypertensive left ventricular hypertrophy.OBJECTIVE: To investigate the relationship between ICAM-1 and hypertensive left ventricular hypertrophy and the effect of tanshinone ⅡA on expression of ICAM-1.DESIGN:A randomized controlled observation was designed.SETTING: Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: The experiment was performed in the Laboratory of Emergency Department of Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from October 2002 to January 2004. Ten male WKY rats(10 WKY rats) of 12-week-old and 20 rats with spontaneous hypertension(20 SHR rats) were employed and divided into the control group(10 WKY rats), hypertension group(10 SHR rats) and tanshinone ⅡA group(10 SHR rats).injected from caudal vein for treatment and distilled water at the same dose was injected in the other two groups. Twelve weeks later, the rats were sacrificed and myocardium was collected for specimen preparation. Haematoxylin and eosin(HE) staining, VG staining, immunocytochemical staining and myocardial ED1 labeling were applied to determine myocardial macrophage infiltration degree. The reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay(ELISA) were used to determine the expressions of ICAM-1 mRNA and protein.MAIN OUTCOME MEASURES: Degree of macrophages infiltration and the expressions of ICAM-1 mRNA and protein in myocardium in each group.RESULTS: Twelve SHR and 10 WKY rats were employed in the experiment pared with the control group, ICAM-1 mRNA and protein expressions in hypertrophic myocardium were increased significantly in hypertension group (0.176±0.087,0.537±0.195;0.104±0.011,0.173±0.027, P ＜0.01or P ＜ 0.05) . Infiltration of macrophage was significant(0. 62 ±0.07,non Ⅱ A group, the expressions of ICAM-1 mRNA and protein were decreased significantly(0. 537 ±0. 195, 0.291 ±0. 106; 0. 173 ±0.027, 0. 125± 0. 014, P ＜ 0.01 or P ＜ 0. 05); the amount of macrophages infiltration was decreased(1.85 ±0. 23, 1.16 ±0. 17, P ＜ 0.05) and the degree of cardiomyocytic hypertrophy and interstitial fibrosis was remarkably relieved.CONCLUSION: Excessive expression of myocardial ICAM-1 and its mediated inflammatory cellular infiltration plays an important role in hypertensive left ventricular hypertrophy. The inhibition of tanshinone ⅡA on left ventricular hypertrophy may be contributed to decreased expression of ICAM-1 and alleviated inflammatory cellular infiltration in myocardium.

OBJECTIVETo explore the effects of integrative medicine (IM) rehabilitation protocolon motor function, activity of daily living, and quality of life (QOL) in hemiplegia patients after stroke.

METHODSTotally 120 patients with post-stroke hemiplegia were allocated to four groups using sealed envalope drawing, i.e., the rehabilitation group, the Chinese medical treatment group, the acupuncture group, and the comprehensive rehabilitation group, 30 cases in each group. Based on routine rehabilitative training, patients in the Chinese medical treatment group, the acupuncture group, and the compre-hensive rehabilitation group received standardized treatment based on syndrome typing, Shi's Consciousness-Restoring Resuscitation acupuncture, Chinese herbs + acupuncture comprehensive rehabilitatino protocol, respectively. The treatmet cycle consisted of 4 weeks with 24-week follow-ups. Fugl-Meyer motor assessment (FMA), Modified Barthel Index (MBI), and Stroke-Specific Quality of Life Scale(SS-QQL), and safety assessment were taken as main effect indices before treatment, at week 4 of treatment, at week 12 and 24 of follow-ups, respectively.

RESULTSThere was no statistical difference in FMA score, MBI score, SS-QOL score among the four groups before treatment (P > 0.05). These scores were significantly improved in the four groups at week 4 of treatment, week 12 and 24 of follow-ups, respectively (P < 0.05). Besides, FMA score and SS-QOL score were significantly improved in the comprehensive rehabilitation group at each corresponding time point, as compared with other treatment groups (P < 0.05).

CONCLUSIONSThe comprehensive protocol could significantly improve motor function, activity of daily living in hemiplegia patients after stroke, and further improve their QOL. Its effect was better than other single treatment.

Activities of Daily Living ; Acupuncture Therapy ; Hemiplegia ; rehabilitation ; Humans ; Integrative Medicine ; methods ; Medicine, Chinese Traditional ; Motor Skills ; Quality of Life ; Stroke Rehabilitation ; Treatment Outcome