OBJECTIVETo study the effect of Flos Daturae alkaloids (FDA) on TGF-beta1-1uuuu;U epithelial-mesenchymal transition (EMT) of human pulmonary adenocarcinoma A549 cells.

METHODSA549 cells in vitro cultured were randomly divided into 5 groups, i.e., the blank control group, the TGF-beta1 group, the low dose FDA group, the medium dose FDA group, and the high dose FDA group. The morphologies of A549 cells were observed. Expressions of cytokeratin (CK)-19 and alpha-smooth muscle actin (alpha-SMA) were detected by Western blot and real-time PCR at 24, 48, and 72 h, respectively.

RESULTSA549 cells in the TGF-beta1, group turned from cobblestone to spindle shape gradually. Those in low, medium and high dose FDA groups showed similar shapes to those of the TGF-beta1 group. There was no statistical difference in the morphology of A549 cells among the 3 dose FDA groups (P > 0.05). Western blot showed that, when compared with the blank control group, the expression of CK-19 was down-regulated, but the expression of alpha-SMA was up-regulated in the TGF-beta1 group (P < 0.01). Compared with the TGF-beta1, group, the expression of CK-19 was up-regulated, but the expression of alpha-SMA was suppressed in low, medium and high dose FDA groups (P < 0.01). The CK-19 expression obviously increased, but the alpha-SMA expression was suppressed in high dose FDA group at 72 h (P < 0.01). Real-time PCR results showed, as compared with the TGF-beta1 group, the mRNA expression of CK-19 was increased, but the mRNA expression of alpha-SMA was reduced in low, medium and high dose FDA groups (P < 0.01).

CONCLUSIONSFDA had no effect on EMT morphological changes of TGF-beta1 induced A549 cells. FDA could reverse characteristic markers of A549 cells during EMT to some extent, such as expressions of CK-19 and alpha-SMA. The expression of CK-19 (as the epithelium marker) increased and the expression of alpha-SMA (as the mesenchymal marker) was reduced. Besides, they were most obviously seen in the high dose FDA group at 72 h in a dose- and time-dependent manner.

Actins ; Adenocarcinoma ; Alkaloids ; pharmacology ; Cell Line, Tumor ; Datura ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; Epithelial-Mesenchymal Transition ; Epithelium ; Humans ; Transforming Growth Factor beta1 ; metabolism

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With the aging of the global population,frailty is now considered a unique old age syndrome,has become an important worldwide public health problems.The elderly people is at increased risk of falls,hip fractures,disability,death and other risks.Motor system disease as an independent risk factor participates in frailty progress and interacts with each other.This article reviews recent studies on frailty and motor system diseases.

Frailty is a geriatric syndrome that has been implicated as a causative and prognostic factor for cardiovascular disease (CVD).Frailty and CVD are often concurrent and mutually promotive.The prevalence of frailty ranges from 10％ to 60％ in patients with CVD,depending on different tools and cutoffs chosen to define frailty.Short-and long-term prognosis of CVD are both affected by frailty.The presence of frailty is correlated with an increase in complications,outpatient and emergency department visits,hospital admissions and stays,and mortality in patients with CVD.Early prevention and clinical intervention can delay or even reverse the development of frailty,thus improving the prognosis for CVD.

In the present study, an ultra performance liquid chromatography coupled with time-of-fight mass spectrometry (UPLC-TOF/MS) based chemical profiling approach was used to evaluate chemical constitution between co-decoction and mixed decoction of ginseng and Radix Aconiti Praeparata. Two different kinds of decoctions, namely co-decoction of ginseng and Radix Aconiti Praeparata: water extract of mixed two herbs, and mixed decoction of ginseng and Radix Aconiti Praeparata: mixed water extract of each individual herbs, were prepared. Batches of these two kinds of decoction samples were subjected to UPLC-TOF/MS analysis. The datasets of t(R) m/z pairs, ion intensities and sample codes were processed with supervised partial least squared discriminant analysis (OPLS-DA) to holistically compare the difference between these two decoction samples. Significant difference between the two decoction samples was showed in the results of positive ion mode. The contents of hypaconitine and deoxyaconitine decreased, while that of benzoylmesaconine, benzoylhypaconine and dehydrated benzoylmesaconine increased in the samples of co-decoction of ginseng and Radix Aconiti Praeparata. The content of diester-diterpenoid alkaloids decreased, while that of monoester-diterpenoid alkaloids increased, which is probably the basis of toxicity-attenuated action when combined ginseng with Radix Aconiti Praeparata.

BACKGROUND:Preliminary studies of our research group mainly focus on the role of nucleus of solitary tract in gastric noxious stimulation and acupuncture point, while dorsal nucleus of vagus nerve, nucleus of solitary tract and area postrema are the necessary component for vagus nerve complex, and dorsal nucleus of vagus nerve plays a crucial role in the regulation of gastric functions. OBJECTIVE:To observe the effect of acupuncture at Zusanli, Neiguan, Pishu and Shenshu acupoints on the gastric-related neurons in dorsal motor nucleus of the vagus nerve. METHODS:The gastric related neurons in dorsal motor nucleus of the vagus nerve of gastric distension rats were identified using extracellular recording technique, according to neuroelectrophysiology method. Acupuncture stimulation was given to Zusanli, Neiguan, Pishu and Shenshu for 15 seconds. The effect of body stimulation on the gastric distension related neurons in dorsal motor nucleus of the vagus nerve of rats was recorded. RESULTS AND CONCLUSION:Among the involved 24 rats, 90 neurons at paraventricular hypothalamic necleus exhibited discharge. Among the discharged neurons, 54 neurons were related to gastric distension. After acupuncture stimulation at Zusanli, Neiguan, Pishu and Shenshu, there were 44, 39, 38, 27 neurons giving a response, the reaction rate was 82%, 72%, 70%and 50%, respectively. Acupunture at different nerve segment acupoints can activate the dorsal motor nucleus of the vagus nerve to different degrees, which is involved in the regulation of gastric motility.

BACKGROUND:Previous studies concerning neuromechanism of acupuncture for regulating gastric function mainly focused on peripheral nerve, involving in central nervous system. In particular, there are few studies addressing higher central nervous system.
OBJECTIVE:To explore the effects of acupuncture at Neiguan (PC6) and Zusanli (ST36) on gastric-related neurons in the hypothalamic paraventricular nucleus.
METHODS:Using microelectrode extracellular recording technique, gastric related neurons in the paraventricular nucleus were found in gastric distension rats. Stimulation was conducted for 30 seconds by hand acupuncture at Neiguan and Zusanli. The effects of acupuncture on gastric-related neurons in the paraventricular nucleus were observed.
RESULTS AND CONCLUSION:The discharge of 109 neurons in the hypothalamic paraventricular nucleus of 60 rats was recorded. A total of 56 gastric-related neurons of the 109 neurons were observed. The number of neurons response to acupunctures at Zusanli, Neiguan, Pishu (BL20) and Weishu (BL21) were respectively 44, 47, 29, and 33, with the reaction proportion of 78.57%, 83.47%, 51.79%and 58.93%. Results suggested that there exist gastric distension stimulation and acupuncture stimulations from neurons soma to visceral reaction in the hypothalamic paraventricular nucleus, which simultaneously participates in the regulation of acupuncture on the gastric function.

Air Pollutants, Occupational ; analysis ; Coke ; Fatty Liver ; epidemiology ; Humans ; Logistic Models ; Male ; Occupational Exposure ; adverse effects ; Risk Factors

Objective To establish the enzyme-linked immunosorbent assay (ELISA) methods for the quantitative determination of IgG antibodies against diphtheria (DT) and tetanus (TT).MethodsPurified diphtheria toxiod and tetanus toxoid were respectively used as the coating antigens,the human-derived serum antibody standard substance of DT and TT served as the standard substance.The dose-response curves of the tested samples and standard substance were fitted.Then the two quantitative ELISA methods for determining the antibody to DT (Anti-DT) and antibody to TT (Anti-TT) were established with the parallel lines method.Then the methodological verification and application study were conducted.Results The validation results of the two quantitative ELISA measurement methods were in accordance with the regulations.The quantity limit of ELISA method for quantitative detection of Anti-DT demonstrated to be 0.084 mIU/mL,its average recovery rate was 97.6%.The intra-assay coefficient of variation(CV) and inter-assay CV of this Anti-DT assay were ≤ 3.40% and ≤5.05%,respectively.The quantity limit of ELISA method for quantitative detection of Anti-TT demonstrated to be 0.175 mIU/mL,its average recovery rate was 97.5%.The intra-assay CV and inter-assay CV of this Anti-TT assay were ≤ 2.42% and ≤5.58%,respectively.These two methods were applied for the immunogenicity evaluation after infantile basic immunization by diphtheria and tetanus vaccines.Conclusion The two established quantitative ELISA methods demonstrate high accuracy and good reproducibility,which are suitable for the ordinary laboratory to carry out the work and can be used in the serological effect evaluation after diphtheria and tetanus vaccine immunization and epidemiological study of diphtheria and tetanus disease.

BACKGROUND: Silibinin has broad pharmaceutical effects, such as anti-free radicals, anti-lipid peroxidation, anti-lipoid oxidase, anti-glutathione (GSH) depletion, anti-neoplastic and serum lipid-lowering effects. Clinically, silibinin is often used in treating alcoholic liver disease. OBJECTIVE: To investigate the pharmacological mechanism of silibinin for alcoholic fatty liver in rats. DESIGN: Randomized and controlled study.SETTING: Guangzhou Hospital of Traditional Chinese Medicine.MATERIALS: The experiment was conducted at the Animal Experimental Laboratory of Guangdong Pharmaceutical Institute from August to October 2003. Totally 57 SD rats, without unusual bacteria, weighting (150±10)g and of either gender, were selected. Yiganling tablets containing 38.5 mg silibinin were produced by Zhuzhou No.3 Pharmaceutical Factory (Batch No. 20020808).METHODS: Among the 57 SD rats, 18 rats were regarded as normal control group. Rats in normal control group were administered with normal saline by gavage, and fed with normal food and distilled water in place of alcohol for 10 weeks. Rats in model group and silibinin group were fed with high-calorie food and 100 mL/L alcohol for 6 weeks to establish model of rat alcoholic fatty liver. The other rats were divided into model control group (n=18) and silibinin group (n=21). Rats in model control group were treated with distilled water while those in silibinin group were treated with 100 mg/kg silibinin. Meanwhile, 100 mL/L ethanol and hyperalimentation feed were given for 4 weeks. After animals were killed, TG, SOD, GSH and MDA levels were measured with liver suspension.MAIN OUTCOME MEASURES: Contents of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP), total cholesterol (TC), triacylglycerol (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), tumor necrosis factor (TNF)-α , and transforming growth factor (TGF)-β1.RESULTS: All the 57 rats entered the final analysis. Silibinin could inhibit the activities of serum AST, ALT and AKP [(2 550.5±400.1), (533.4±100.0), (2 217.1±750.2)nkat/L], and the differences were significant as compared with those in model control group [(3 600.7±666.8), (800.2±100.0), (2 900.6±1 333.6) nkat/L, P ＜ 0.05-0.01]. Contents of TG, LDL-C, TNF-α and TGF-β1 in silibinin group [(1.8±0.8), (0.17±0.04), (6.66±1.38), (24.1±4.1) mmol/L] were lower than those in model group [(2.8±1.4), (0.20±0.05), (7.81±1.06), (28.8±6.3) mmol/L] with significant differences (P ＜ 0.05-0.01). Silibinin could increase the content of HDL-C but decrease the contents of TG and MDA (P ＜ 0.05-0.01), and improve SOD activity as well as hepatocyte and fatty degeneration (P ＜ 0.01).However, it had no obvious effect on the content of reduced estathion (P ＞ 0.05).CONCLUSION: Silibinin can inhibitthe formation of alcoholic fatty liver in rats. The pharmacological mechanism of silibinin may involve anti-oxidation, removing free radicals, inhibiting lipid peroxidation, regulating blood lipid component, reducing fatty sediment in liver, and anti-immunoinflammation and anti-hyperplasia effects.