汤剂煎煮-剂数-服用频次·时机与疗效

汤剂服法是辨证施治重要步骤,对提高服药质量,提升临床疗效,具重要意义和实用价值。因病定剂:依疾病势态发展确定服药剂数,病情轻缓,病势不急,每日一剂;病程较长,不能急攻或药物具有烈性和毒性,易损伤正气,宜两日一剂;病情急重或病势凶猛,须截断扭转,缓和病势,或药剂轻清,不宜久煎,病势又须即时控制,则用同一处方,每日二剂或三剂以截病势;病情复杂,正虚邪实,攻之伤正,补之碍邪,攻补同用,病又难除,可双处方交替使用,既攻邪势,又扶正气。因病定次:病情急、邪势重,需急挫邪势,缓解病情,或药力集中,扭转病势,或轻药不得重用,或峻药慎用,顿服或日一服;病情不急,病势不凶,需药力接续,常日服二、三次;儿科病,起病急,发展快,服药较难,不易达到和维持血药浓度及有效剂量,宜小量多次服;需药物充分接触患处,饮之不及,宜含咽之法。因病定量:平均等量,适用于病情变化小,病势不急;药量渐增,以防不良反应,中病即止;集中服量,急救时立即服用,以求即刻扭转病势;视体质服量,防攻伐太过,损伤正气;试病服量,以防过剂给药;探病服量,以便确定治疗方案,依病严控,以知为度,中病即止,渐增渐减,积累获效。因病定服:病有寒热上下,药有温热寒凉,根据疾病性质、部位,"病在胸膈以上者,先食而后服药,病在胸腹以下者,先服药而后食,病在四肢血脉者,宜空腹而在旦,病在骨髓者,宜饱满而在夜"。

Real-Time PCR Technique and Its Application in Quantification of Plant Nucleic Acid Molecules

Real-time PCR is a closed DNA amplification system that skillfully integrates biochemical, photoelectric and computer techniques. Fluorescence data acquired once per cycle provides rapid absolute quantification of initial template copy numbers as PCR products are generated. This technique significantly simplifies and accelerates the process of producing reproducible quantification of nucleic acid molecules. It not only is a sensitive, accurate and rapid quantitative method, but it also provides an easier way to calculate the absolute starting copy number of nucleic acid molecules to be tested. Together with molecular bio-techniques, like microarray, real-time PCR will play a very important role in many aspects of molecular life science such as functional gene analysis and disease molecular diagnostics. This review introduces the detailed principles and application of the real-time PCR technique, describes a recently developed system for exact quantification of AUX/IAA genes In Arabidopsis, and discusses the problems with the real-time PCR process.

Quantitative analysis of transforming growth factor beta 1 mRNA in patients with alcoholic liver disease

AIM: To investigate the expression of the transforming growth factor beta 1(TGF-beta 1) mRNA in different stages of alcoholic liver disease (ALD) and its clinical value. METHODS: One hundred and seven male alcoholics were grouped by clinical findings into four groups: alcohol abusers without liver impairment (n =22), alcoholic steatosis (n =30); alcoholic hepatitis (n=31); and alcoholic cirrhosis(n=24). Using peripheral blood mononuclear cells (PBMC) as samples the gene expression of TGF-beta 1 was examined quantitatively by reverse transcription polymerase chain reaction (RT-PCR) and dot blot. There are 34 healthy subjects served as control. RESULTS: The expression of TGF-beta 1 from all ALD patients was significantly greater than that in controls (1.320 +/- 1.162 vs 0.808 +/- 0.276, P【0.001). The differences of the expressions were significant between the patients from each groups (alcoholic steatosis, alcoholic hepatitis and alcoholic cirrhosis) and the controls (1.168 +/- 0.852, 1.462 +/- 1.657, 1.329 +/- 0.610 vs 0.808 +/- 0.276, P【0.050). No significant differences of TGF -beta 1 mRNA expression were observed between alcohol abusers without liver impairment and controls. The expressions in patients with alcoholic hepatitis and alcoholic cirrhosis were significantly greater than that in alcohol abusers respectively (1.462 +/- 1.657, 1.329 +/- 0.610 vs 0.841 +/- 0.706, P【0.050). No significant differences of TGF-beta 1 mRNA expression were observed between alcoholic fatty liver men and alcohol abusers. CONCLUSION: TGF-beta 1 expression level can be a risk factor for alcoholic liver disease and might be related to the inflammatory activity and fibrosis of the liver in patients.

Function of nonstructural 5A protein of genotype 2a in replication and infection of HCV with gene substitution

AIM:To explore the function of Nonstructural 5A(NS5A) protein of genotype 2a(JFH1)in the replication and infection of hepatitis C virus(HCV).METHODS:Intergenotypic chimera FL-J6JFH/J4NS5A was constructed by inserting NS5A gene from 1b stain HC-J4 by the overlapping polymerase chain reaction (PCR)method and the restriction enzyme reaction.In vitro RNA transcripts of chimera,prototype J6JFH and negative control J6JFH1(GND)were prepared and transfected into Huh-7.5 cells with liposomes.Immunofluorescence assay(IFA),fluorescence quantitative PCR and infection assay were performed to determine the protein expression and gene replication in Huh-7.5 cells.RESULTS:The HCV RNA levels in FL-J6JFH/J4NS5A chimera RNA transfected cells were significantly lower than the wild type at any indicated time point(2.58 ±5.97×106 vs 4.27±1.72×104,P=0.032).The maximal level of HCV RNA in chimera was 5.6±1.8× 104 GE/μg RNA at day 34 after transfection,while the wild type reached a peak level at day 13 which was 126 folds higher(70.65±14.11×105 vs 0.56±0.90 ×105,P=0.028).HCV proteins could also be detected by IFA in chimera-transfected cells with an obviously low level.Infection assay showed that FL-J6JFH/J4NS5A chimera could produce infectious virus particles,ranging from 10±5 ffu/mL to 78.3±23.6 ffu/mL,while that of FL-J6JFH1 ranged from 5.8±1.5×102 ffu/mL to 2.5±1.4×104 ffu/mL.CONCLUSION:JFH1 NS5A might play an important role in the robust replication of J6JFH1.The establishment of FL-J6JFH/J4NS5A provided a useful platform for studying the function of other proteins of HCV.

Expression of the B-Cell Lymphoma/Leukemia 11A Gene in Malignant Hematological Cell Lines through Quantitative Reverse Transcription Polymerase Chain Reaction

The B-cell lymphoma/leukemia 11A （BCL11A） gene is essential for normal lymphoid development and has been associated with hematological malignancies. In the current study, the relative expression level of BCL11A in malignant hematological cell lines was evaluated through real-time quantitative reverse transcription polymerase chain reaction （qRT-PCR）. METHODS The relative expression level of BCLllA mRNA in malignant hematological cell lines was determined through qRT- PCR using SYBR Green I dye. Glyceraldehyde-3-phosphate dehydro- genase was used as the reference gene to confirm the relative expression level of BCL11A gene mRNA. RESULTS The relative expression level of BCL11A mRNA in cell lines from B-cell malignancies was significantly higher compared with that from acute rnyeloid leukemia （P 〈 0.05）. Different cell lines with malignant B-cells exhibited a wide range of BCL11A expressions ranging from 27.37 to 93.38. CONCLUSION The overexpression of BCL11A gene mRNA in malignant B-cells might play a role in B-cell lymphoma/leukemia.

A Quantitative Assay for Measuring of Bovine Immunodeficiency Virus Using a Luciferase-based Indicator Cell Line

In order to quantitate the bovine immunodeficiency virus line （BIVL） was established by transfecting baby hamster kidney （BIV） cells infection in vitro, a BIV indicator cell with reporter plasmids containing the firefly luciferase gene driven by a BIV long terminal repeat promoter. The BIV activates promoter activity of the LTR to express luciferase upon infection. BIV infection could therefore by quantified by detection of luciferase activity. Compared to standard assays used to detect BIV infection, the BIVL-based assay is 10 times more sensitive than the the CPE-based assay, and has similar sensitivity with the viral capsid protein Western blot assay BIV indicator cell line could detect BIV infection specifically. Luciferase activity of BIV infected BIVL cells showed a time dependent manner, and 60 h post infection is the optimal time to detect BIV infection. Luciferase activity of BIVL cells correlates with the BIV capsid protein expression. Moreover, a linear relationship was found between MOI and the activated intensity of luciferase expression. In brief, the BIV indicator cell line is an easy, robust and quantitive method for monitoring BIV infection.

What the Hell Am I Doing Here？ A Look at a Series of Geriatric Centers, Profile of the Institutionalized Persons and Reasons for Admission

The main objective of this study was always to know the profile of the institutionalized people in our environment, to know their reality： age, age of admission, level of dependency and evolution during their stay, years of stay, number of children, reason for admission. We use data from more than 600 patients from different residences of different backgrounds： public and private, lay and religious, rural and urban. We performed a descriptive study expressing the results in percentages with standard deviation and later evaluated the statistical significance of the differences using the student＇s t-test for the quantitative and Chi-square variables to compare qualitative variables. The results of the study are numerous and of diverse nature, because of their extension, from the general profile of the resident, to the important gender differences, attributed in principle to the different roles of each gender in the studied generations. There are also differences depending on the funding, the reason for admission or the environment. This is at the end only the beginning of a large comparative study with non-institutionalized population, in order to compare this population.

Expression of pathogenesis-related genes in cotton roots in response to Verticillium dahliae PAMP molecules

Verticillium wilt disease becomes a major threat to many economically important crops. It is unclear whether and how plant immunity takes place during cotton-Verticillium interaction due to the lack of marker genes. Taking advantage of cotton （Gossypium hirsutum） genome, we discovered pathogenesis-related （PR） gene families, which have been widely used as markers of immune responses in plants. To profile the expression of G. hirsutum PR genes in the process of plant immunity, we treated cotton roots with two immunogenic peptides, fig22 and nlp20 known as pathogen-associated molecular patterns, as well as three Verticillium dahliae-derived peptides, nlp20vd2, nlp23vd3, and nlp23vd4 which are highly identical to nlp20. Quantitative real-time PCR results revealed that 14 G. hirsutum PR gene （GhPR） families were induced or suppressed independently in response to fig22, nip20, nlp20va2, nlp23vd3, and nlp23vd4. Most GhPR genes are expressed highest at 3 h post incubation of immunogenic peptides. Compared to fig22 and nlp20, nlp20vd2 is more effective to trigger up-regulated expression of GhPR genes. Notably, both nlp23vd3 and nlp23vd4 are able to induce GhPR gene up-regulation, although they do not induce necrosis on cotton leaves. Thus, our results provide marker genes and new immunogenic peptides for further investigation of cotton-V, dahliae interaction.