Rhizome Research Articles

Drug analysis of Li zhenhua for the treatment of chronic atrophic gastritis based on the data mining

Objective The method of data mining and sorting analysis was used to analyze and summarize the drug experience of Professor Li Zhenhua in the treatment of chronic atrophic gastritis (CAG). Methods Professor Li's medical records of effective diagnosis and treatment of 139 CAG patients were collected. Find-Replace method of Microsoft office word 2007 was used to count the major syndromes and main prescription of CAG. SPSS statistical software was adopted to perform entry-analysis-descriptive statistics and data analysis of and main syndrome, main formula and frequency of administration so as to obtain the commonly used drugs, commonly used prescription and drug laws of CAG. Results Professor Li Zhenhua believed that the clinical syndromes of CAG included the disharmony of liver-stomach-spleen syndrome, the damp heat of spleen-stomach syndrome, the deficiency and damp heat of spleen syndrome, the liver depression and spleen deficiency syndrome, the deficiency of spleen-stomach syndrome, the liver and stomach yin deficiency and qi stagnation syndrome, the stagnant heat of liver-stomach syndrome and the blood stasis of stomach meridian syndrome; the commonly used drugs were: bupleurum, white peony root, orange peel, licorice, poria, skullcap, ginger, fried atractylodes, golden thread, prepared pinellia, licorice, lily, stir-baking Sanxian, nutgrass galingale rhizome, heterophylly falsesatarwort root, combined spicebush root, Chinese date, tangshen, immature orange fruit, prepared rhizome pinellize without adjuvant, and oyster shell..The commonly used prescriptions were: Xiaochaihu decoction, Sini powder, Chaihu-Guizhi-Longgu-Muli decoction, Chaihu-Shugan powder, Huanglian-Wendan decoction, Banxia-Xiexin decoction, Xiaoyao powder, Xiangsha-Liujunzi decoction. Conclusion Professor Li pay attention to treat spleen and stomach disease from liver by clearing heat and removing dampness from spleen and stomach. He used the dialectical methods like invigorating qi and strengthening the spleen, regulating qi digestion, activating blood flow to eliminating blood stasis. Key words: Chronic atrophic gastritis; Expert experience

After the rhizome

After the rhizome

An Analysis of Contact Improvisation as Rhizome

An Analysis of Contact Improvisation as Rhizome

Content and antioxidant activity of total polyphenols and tannins from Athyrium multidentatum （Doll.）Ching

Objective To determine the content and to evaluate the antioxidant activity of total polyphenols and tannins from Athyrium multidentatum（Doll.）Ching rhizome. Methods Folin phenol reagent-casein method was used to determine the content of total polyphenols and tannins. Spectrophotometry was used to evaluate the antioxidant activity. Results The content of total polyphenols and tannins was 24.61%and 4.59%, respectively. The total polyphenols and tannins exhibited strong reducing power and hydroxyl radicals/superoxide radicals scavenging capability. Conclusion The antioxidant activity of total polyphenols and tannins from Athyrium multidentatum（Doll.）Ching rhizome is significant and further investigation is deserved in the future. Key words: Athyrium multiclentatum （Doll.） Ching; Tannins; Polyphenols; Antioxidants （TCD）Spectrophotometry

Nutritional Evaluation of some Edible Underground Plant Parts Used by the Tribal People of South Odisha, India

The paper deals with the nutrient analysis of eleven edible underground plant parts of South Odisha, India, most of which are used by the tribal/ rural people at the time of food scarcity. The nutritive value such as crude protein, total sugar, total lipid, vitamin B and vitamin C content of the selected edible underground plant parts were analysed. The plants with edible underground parts collected for nutritional analysis are Amorphophallus paeoniifolius (Dennst.) Nicolson (Amorphophallus campanulatus Decne.), Colocasia esculenta (L. ) Schott, Curcuma angustifolia Roxb., Dioscorea alata L., D. oppositifolia L., D . wallichii Hook. f. (bulbil and rhizome), Musa × paradisiaca L., Nelumbo nucifera Gaertn., Nymphaea nouchali Burm. f., Raphanus raphanistrum subsp. sativus (L.) Domin ( Raphanus sativus L.).

Identification of Marker Nucleotides for the Molecular Authentication of Arisaematis Rhizoma Based on the DNA Barcode Sequences

ABSTRACT Objectives : Official Arisaematis Rhizoma is described only three species, Arisaema amurnse , Arisaema erubescens , and Arisaema heterophyllum, in national Pharmacopoeia. However, other Arisaema species, Arisaema ringens, Arisaema takesimense and Arisaema serratum, also have been distributed as an inauthentic Arisaematis Rhizoma in the herbal market. To develop a reliable molecular a uthentication method for Arisaematis Rhizoma in species level, we analyzed DNA barcode regions using six Arisaema species.Methods : Thirty-eight samples of six Arisaema plants species (A. amurense, A. amurense f. serratum, A. heterophyllum, A. takesimense, and A. serratum) were collected from different habitate and nucleotide sequences of DNA barcode regions (rDNA-ITS, matK, and rbcL gene) were analyzed after PCR amplification. The species-spec ific sequences and phylogenetic relations were estimated using entire sequence s of three DNA barcodes based on the analysis of ClastalW and UPGMA, respectively.Results : The comparative analysis of DNA barcode sequences were revealed inter-species specific nucleotides to distinguish the medicinal plant of Arisaema Rhizoma in species levels excluding between A. amurense and its subspecies (A. amurense f. serratum) and A. takesimense and A. serratum, respectively. However, we obtained sequence differences enough to discriminate authentic and inaut hentic Arisaematis Rhizoma. Therefore, we suggest that these SNP type molecular genetic markers were an r eliable method avaliable to identify official herbal medicines.Conclusions : These marker nucleotides could be useful to identify the official herbal medicines by providing definitive information that can identify original medicinal pla nt and distinguish from inauthentic adulterants and substitutes.Key words : Arisaema species, Arisaematis Rhizoma, DNA barcode, Marker Nucleotide, Molecular Authentication

A study of functional components antioxidant activity and sensory characteristics of Gastrodiae Rhizoma by steaming-drying cycles

ABSTRACT Objectives : The objective of this study is to determine the best numbers o f steaming-drying cycles of Gastrodiae Rhizoma for both efficacy and taste. We investigated various characteristics among Gastrodiae Rhizoma samples (GSD1, GSD3, GSD5 and GSD7) through the number of steaming-dryi ng cycles increased.Methods : Gastrodiae Rhizoma were steamed and dried at different repeated numbers. They wer e divided into samples (GSD1; steamed and dried once, GSD3; steamed and dried three times, GSD5; steamed and dried five times, GSD7; steamed and dried seven times) for experiment. The y were extracted using water, freeze dried and powdered to analyze proximate composition, free sugar amount, f unctional components, antioxidant activity and sensory evaluation.Results : Proximate composition and the amount of free sugars of Gastrodiae Rhizoma did not have meaningful differences among samples. Phenolic and flavonoid content of sa mples were increased by increasing steaming-drying numbers. Gastrodin content had different values and GSD7 was th e highest in comparison with others. Increasing steaming-drying numbers led to a increasing in radic al and nitrate scavenging activity in samples. Regarding to sensory evaluation, GSD5 was selected as the best sample according to its highest hedonic score mean (5.54/7) among all samples for appearance, color, taste an d overall acceptability.Conclusions : The results indicated that 5th-cycling sample was effective in views of functional components, antioxidant activity and sensory characteristics. Moreover, it was suggested that steaming-drying process improved remarkably the effects of Gastrodiae Rhizoma.Key words : Gastrodiae Rhizoma, steaming-drying cycles, gastrodin, antioxidant activity, sens ory characteristics

Liver protective effects of Siphonostegia chinensis decoction in rats with liver fibrosis

Objective To investigate the liver protective effects of Siphonostegia chinensis decoction in rats with liver fibrosis induced by carbon tetrachloride（CCl4）.Methods A total of 120 male SD rats were randomly divided into six groups： a blank control group, a model group, a Siphonostegia chinensis decoction group, a Salvia miltiorrhiza decoction group, a Rhizoma polygoni cuspidati decoction group and a polyene phosphatidylcholine capsules group, with 20 rats in each group. The model of liver fibrosis in rats was induced by subcutaneous injection with CCl4, except the blank group. Except rats in the blank control group and the model group, the rats in the other four groups were treated with Siphonostegia chinensis decoction, Salvia miltiorrhiza decoction, Rhizoma polygoni cuspidati decoction and polyene phosphatidylcholine capsules（10 ml/kg）. The serum level of tumor necrosis factor-α（TNF-α）was determined with enzyme-linked immunosorbent assay, the fibrosis stage assessment were determined by histopathological examination.Results Compared with the model group, Siphonostegia chinensis decoction group significantly reduced the stage of liver fibrosis（P〈0.01）and there were no obvious difference in liver fibrosis stage among the Salvia miltiorrhiza decoction group, Rhizoma polygoni cuspidati decoction group and polyene phosphatidylcholine capsules group, and the Siphonostegia chinensis decoction group（allP〉0.05）.Compared with liver fibrosis model group（368.06±24.90）U/L, the serum level of TNF-α in the Siphonostegia chinensis decoction group（336.61± 20.20）U/L significantly reduced（P〈0.01）, and there were no obvious difference among the Siphonostegia chinensis decoction group, the Salvia miltiorrhiza decoction group（337.81±21.04）U/L, the Rhizoma polygoni cuspidati decoction group（338.95±24.43）U/L and the polyene phosphatidylcholine capsules group（337.11± 23.64） U/L（allP〉0.05）.Conclusion Siphonostegia chinensis decoction has certain liver protective effect in rats with liver fibrosis induced by CCl4. Key words: Siphonostegia chinensis; Liver Fibrosis; Drug-Induced Liver Injury; Carbon Tetrachloride

Effects of Imperatae Rhizoma Extract on T helper 2 cell differentiation

Objectives : The aims of this study were to exploring the therapeutic effect of Imperatae Rhizoma Extract(IRE) on Asthma. Methods : To investigate biological modulation activities of IRE, we conducted the cell-based assay whether IRE could regulate T helper 2 cells activity with EL4 T cells and mouse splenocytes, and followed animal study to conform the efficacy of their therapeutic potential on OVA-induced asthmatic mouse. Results : In cell study, IRE suppress the nuclear translocation of GATA binding protein-3 protein in phorbol 12-myristate 13-acetate/Ionomycin-stimulated EL4 T cells and Interleukine(IL)-4, IL-5 and IL-13 production in splenocytes at concentration dependent manner. In animal study, IRE-treated groups both 100mg/ml and 200mg/ml improve airway hypersensitibility reaction(AHR) response to methacholine about 30% and 40% with positive control group. Peritoneal blood analysis reveal that eosinophil number and ovalbumin-specific IgE is reduced by IRE treatment. Cell number of eosinophil is also reduced in bronchoalveolar lavage of IRE group like to peritoneal cell and real time-polymerase chain reaction data show that expression levels of IL-4, IL-5 and IL-13 were down regulated in lung tissue. Finally, histological analysis indicate that IRE protect the bronchial tissue damages through the accumulation of inflammatory cells and collagen, and these effect may be cause by interfering Th2 cells activity. Conclusions : Our data represent that IRE potentiates therapeutic activities to the allergic diseases such as asthma by regulating Th2 cells differentiation.

Inhibition Effect of Gamisoyo-san on MITF, TRP-1, TRP-2, Tyrosinase mRNA Expression in Melanoma Cells (B16F10)

ABSTRACT Objectives : Gamisoyo-san complex prescription were made with Angelicae Gig antis Radix, Paeoniae Radix, Atractylodes rhizome white, Hoelen, Bupleuri Radix, Moutan Cortex Radicis, Gardeniae Fructus, Zingiberis Rhizoma Crudus, Menthae Herba. The purpose of this study was to research the whitening effect of the extract from Gamisoyo-san, which is one of the used herbal complex prescription.Methods : This study investigated inhibitory effect of Gamisoyo-san in t yrosinase activity. Cell viability were performed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Then, Gamisoyo-san measured reversed-transcription-PCR for mRNA expression using B 16F10 mouse melanoma cells.Results : For whitening effects, the tyrosinase inhibition effect of extract was shown to 52.4% at 5,000㎍/㎖ concentration. The cell viability on B16F10 melanoma cells of G amisoyo-san extract showed higher than 75% at 1,000㎍/㎖ concentration. In this study, an experiment was performed b y setting the non-toxic concentration range of 50, 150, 250㎍/㎖. The Glyceraldehyde-3-phosphate dehydrogenase ( GAPDH) was used as a positive control. The microphthalmia-associated transcription factor (MITF), tyrosina se related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), tyrosinase mRNA expression inhibitory by re verse transcription-PCR of Gamisoyo-san extract were decreased by 95.3%, 98.8%, 96.3% and 49.5% at 250㎍/㎖ which the highest concentration.Conclusions : All these findings could verify that whitening effects of Gamisoyo-san extract by tyrosinase inhibitory activity and mRNA expression. The Gamisoyo-san could be used as material for functional cosmetics, such as skin whitening products.Key words : Gamisoyo-san, MITF, TRP-1, TRP-2, tyrosinase

Composition of the essential oils of three Cyperus species from Congo

Essential oils from underground parts of Cyperus articulatus (L.), Cyperus sp. and Kyllinga brevifolia (S.) were analyzed by gas chromatography mass spectrometry (GC-MS). Thirty four (34) components were identified in the oil of C. articulatus. The main components were α-cadinol (12.07%), trans-pinocarveol (9.86%), cyperenone (7.28%), cyperene (6.29%), myrtenol (5.50%), myrtenal (4.90%) and cis-carveol (4.42%). Fifteen (15) components were identified in the oil of Cyperus sp. The main components were farnesol (28.61%), humulene epoxide (12.86%) and caryophyllene oxide (8.96%). Twenty three components were identified in the oil of K. brevifolia with the main components being manoyl oxide (44.08%), β-pinene (13.58%), cyperene (7.63%) and γ-terpineol (7.37%). Only β-pinene and limonene were present in the three species of Cyperaceae. The findings show broad differences in chemical compositions of the essential oils of C. articulatus, Cyperus sp. and K. brevifolia. When compared with previous studies available in literature, it was also noticed that a difference of composition existed for oils of C. articulatus and K. brevifolia of other origins, respectively. Key words: Cyperus articulatus, Cyperus sp., Kyllinga brevifolia, Cyperaceae, essential oil, rhizome.

Effect of ginger rhizome extract on lymphocytopenic guinea pig with azathioprine

The objectives of this study were to investigate quantitative alterations in leukocytes and immunosuppressive effects of ginger rhizome associated with oral administration of Azathioprine (AZA) on guinea pig model. This study was carried out at the Al Jouf University during the period from March to April 2013. A total of twelve guinea pigs randomly divided into three groups (four guinea pigs each) were used in this study. Group I, received daily oral administration of 50 mg/kg body weight of AZA in combination with 50 mg/ml of ginger rhizome extract for five days; Group II was treated with 50 mg AZA daily for five days, while Group III was used as a control group. Hematological alterations associated with ginger rhizome and AZA were measured using standard hematological and data analysis techniques. Ginger rhizome in combination with AZA was found to enhance the reduction in the lymph (%) compared to AZA treatment (P < 0.05). Lymphocyte was significantly lower in group II (AZA) and group I (AZA + ginger) (P < 0.05), 13 and 18%, respectively than the control group, 38%. This may be due to either inhibition in one of the lymphopoiesis growth factors or shortening the life span of lymphocytic cells by the action of AZA. The results of the present study indicated an immune suppressive effect of ginger rhizome extract associated with immunosuppressive agent (AZA) with mainly lymphocytopenia. The current findings suggest that ginger may have a role in either the stimulation of lymphopoiesis or discontinue the effect of AZA, as the percentage of lymphocytes was significantly higher in group II versus group I after mixing up AZA + ginger. Accordingly, further studies would be useful to estimate lymphocytic growth factors to know the relationship between ginger, AZA and lymphopoiesis. Key words: Azathioprine, ginger rhizome, blood cell counting, hematological alterations.

Bacteriostatic or bactericidal action of four aqueous plant extracts on multi-drug resistant bacteremia and their effect on cells morphology recorded using scanning electron microscopy (SEM)

Four aqueous traditional plant extracts from rosemary leaves (Rosmarinus officinalis), clove buds (Syzygium aromaticum [L.] Merr at Perry, Myrtaceae), cinnamon bark (Cinnamomum zeylanicum) and ginger rhizomes (Zingiber officinale) were tested as natural antibiotics against multi-drug resistant bacteremia isolates: Six Gram negative isolates, viz. Acinetobacter baumanni/haemolyticus, Citrobacter freundii, Escherichia coli, E. coli ESβL, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella sp. and four Gram positive isolates viz. methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus epidermidis, Streptococcus pneumoniae and Bacillus sp. were which used to test antibiotic activity. The usage of clove extract (25X) appeared to have maximum antibacterial activity against all tested Gram negative bacteria, when compared with other tested plant extracts at the same concentration. In the case of G+ve bacteria, tested extracts appeared to have almost the same efficiency. Swabs were taken from inhibition zone resulting from the inhibitory effect of aqueous extracts towards the tested isolates and re-inoculated to detect either the bacteriostatic or bactericidal action. Again, clove extract exhibited the highest effect when compared with other tested against the isolates from bloodstream. Scanning electron microscopy was used to record external morphological changes of the bacterial cells such as appendages, shortness and lyses. Key words: Aqueous plants extracts, bacteremia, antibiotics, rosemary, clove, cinnamon, ginger, Escherichia coli, Salmonella, Acinetobacter, Citrobacter, Klebsiella, Pseudomonas, Streptococcus, Bacillus, Staphylococcus.

A Study on “becoming-” process as Rhizome’s breakaway －in text 《Avatar》－

A Study on “becoming-” process as Rhizome’s breakaway －in text 《Avatar》－

NON TIMBER FOREST PRODUCTS UTILIZATION DURING ADVERSE CLIMATIC CONDITIONS : A CASE OF SAORA TRIBES OF GANJAM DISTRICT, ODISHA

Saoras are one of the most ancient tribes in Odisha mentioned in Hindu myths and classics are also expert climbers and hunters. Saoras of Ganjam district mostly depend on forest for their livelihood and the area is rich in biodiversity and also have rare medicinal flora. They eat several NTFPs, wild roots, rhizome, seeds, fruits and mushrooms during harsh climatic conditions like severe drought. They use stored wild tubers mostly Dioscoreas like Bowla ( Dioscorea bulbifera L.), Bonda, Chun aru ( D. daemona Roxb.), Karondi aru ( D.bellophylla L.), Pani aru ( D. oppositifolia L.) and Arguna ( Cycas spp. ) to fulfill their food, medicinal and nutritional requirements particularly in harsh climatic conditions. They eat various wild leafy vegetables like Kolod ( Lathyrus sativa L.),Bilo ( Pisum sativum L.), Kulthi ( Microtylum uniflorum Lam.), Banana stem ( Musa superb a L.) Ban poi ( Basella alba L.). They also utilize NTFPs like Bamboo karda ( Bambusa vulgaris Schrad exJC Wendl.), Moha ( Madhuca indica J.F Gmel.), Amla ( Emblica officinalis Gaertn.), fruits. Some wild mushrooms like Banschattu, Ambachattu ( Agaricus spp. ) after drying and crushing were eaten by them. Indigenous practices of Saora tribes to cope up extreme environmental conditions and utilization of forest products during drought were discussed in the paper.

Curcumin ameliorates gastrointestinal dysfunction and oxidative damage in diabetic rats

Diabetes is known to be associated with gastrointestinal complications characterized by nausea, vomiting, early satiety, bloating, and abdominal discomfort or pain commonly occurring in the advanced stages of the disease. Curcumin is the lipid-soluble antioxidant obtained from the rhizomes of Curcuma longa Linn, also known as turmeric. Curcumin targets multiple chemotherapeutic and oxidative stress pathways and has demonstrated safety and tolerability in humans, supporting its potential as a therapeutic agent; however, literature lacks conclusive evidence supporting its use as a therapeutic agent for the treatment of diabetes induced gastrointestinal complications. Hence, Curcumin was given in different doses to SD rats after 4 weeks of diabetic GI complication induction. At the end of 4 weeks, significant GI dysfunction characterized by weight loss, delayed gastric emptying and intestinal transit associated with reduction in antioxidant enzyme levels and increased lipid peroxidation was observed. Upon treatment with Curcumin for further 4 weeks, reversal of GI dysfunction evidenced by restoration of body weight, GI emptying, intestinal transit, and restoration of antioxidant enzyme level and lipid peroxidation proves the beneficial role of Curcumin in diabetes induced GI complications due to its antioxidant potential.

BIOASSAY DIRECTED FRACTIONATION AND IDENTIFICATION OF BIOACTIVE COMPONENTS FROM ROOT EXTRACT FROM PIPER LONGUM

Piper longum L. (Piperaceae) commonly known as “long pepper” is a well-known medicinal plant in traditional and Ayurveda system. Different parts of this plant, such as root, seed, fruit, whole plant etc. are used traditionally in various ailments. In the present study, we have investigated the antimicrobial activity of ethanol and methanol extract from P. longum rhizome against major skin pathogens Staphylococccus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus vulgaris, and Proteus mirabilis . The most promising activity of methanol extract was observed against Klebsiella pneumonia, Proteus vulgaris, and Staphylococccus aureus with zone of inhibition of 17mm diameter. Further bioactivity-guided fractionation of methanol extract by silica gel column chromatography resulted in eleven major fractions. Among these, fraction-1, 4, 5 and 6 showed higher antimicrobial activity against selected skin pathogens. The bioactive fraction of the methanol extract fraction-5 was subjected for further bio analytical studies. The presence of various components as alkaloids, trepenes, and flavonoids were confirmed after phyto-chemical analysis. The GC-MS analysis of the bioactive fraction identified the major compounds as Piperine, piperlogume, and other components in traces along with some unidentified constituents. The data obtained confirmed its promising bioactivity and led a basis for its significant use as a therapeutic, both in traditional and folk medicine.

Analysis of five bioactive compounds from naturally occurring Rhodiola rosea in eastern Canada

Saunders, D., Poppleton, D., Struchkov, A. and Ireland, R. 2014. Analysis of five bioactive compounds from naturally occurring Rhodiola rosea in eastern Canada. Can. J. Plant Sci. 94: 741-748. The plant Rhodiola rosea produces several bioactive compounds including salidroside, tyrosol, rosarin, rosavin, and rosin. These compounds have been used in traditional medicinal practices in Asia and are currently found in many nutraceutical mixtures. The nutraceutical qualities of these five compounds has resulted in over harvesting of natural populations of R. rosea and instigated several industrial cultivation operations. The purpose of this study was to examine R. rosea from five distinct natural populations in the maritime provinces of Canada to determine the concentrations of these five bioactive compounds. Ten samples were obtained from each of five sites and the concentration of each compound in the rhizomes was determined. Reverse phase-HPLC with a diode array detector was used to quantify the five compoun...

Analysis the outcomes of traditional Chinese medicine clinical pathway in the treatment of cervical spondylotic radiculopathy

Objective To investigate the effect of treating cervical spondylotic radiculopathy with traditional Chinese medicine. Methods 110 patients with cervical spondylotic radiculopathy were treated with traditional Chinese medical pathway： Chinese herbal medicine soaking （incised notoptetygium rhizome, Chinese angelica, divaricate saposhnikovia root, safflower, dahurian angelica root, myrrh, common floweringqince fruit, and tuberculate speranskia herb, etc.）-Acupuncture-cervical traction-massage, once daily, 10 times as a course of treatment. After two courses, therapeutic effect score was analyzed. Results Of all 110 cases, 35 cases（31.82%）were cured; 53 cases（48.18%）had obvious effectiveness; 17 cases（15.45%）had effectiveness; 5 cases（4.55%）had no effectiveness. The total efficiency was 95.45%. The patient’s age（χ2=0.296）, sex（U=0.023）, and duration of disease（χ2=0.577）had no significant effect on the efficacy（P>0.05）;After the treatment, neck pain, upper extremity pain, upper extremity numbness, neck tenderness, intervertebral foramina compression test, sensory disturbances limbs, myodynamia of upper limbs, and tendon reflexes were significantly improved compared with before treatment, with significant difference（t=16.355、11.958、7.755、4.927、11.780、8.647、10.485、3.190、3.873、3.367,P<0.05）. Conclusion The use of traditional Chinese medicine clinical pathway in treating cervical spondylotic radiculopathy was effective and it could improve the quality of medical care. Key words: Cervical spondylotic radiculopathy; Traditional Chinese medicine clinical pathway; Clinical efficacy

Effects of traditional Chinese drugs on cultured murine CD8+ cytotoxic T lymphocytes

Objective To establish a model for studying CD8+ cytotoxic T lymphocyte proliferation in vitro and to screen traditional Chinese drugs (TCDs) with immunosuppressive effects.Methods Spleen tissue was isolated from mice,and made into single cell suspensions followed by separation of CD8+ T lymphocytes with specific antibodies.Then,the CD8 + T lymphocytes were seeded into anti-CD3/CD28 antibody-coated 96-well plates and cocultured with the extracts of 23 TCDs (100 mg/L) separately for 96 hours.Those ceils cultured with and without the presence of anti-CD3/CD28 antibody alone served as the positive control and negative control respectively.The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,inner salt (MTS) assay was performed to evaluate the proliferation of cells and to select the top four TCDs with the strongest inhibitory effect.The relationship between the inhibitory effect and TCD concentrations was further assessed for the four selected TCDs.Enzyme-linked immunospot (ELISPOT) assay was carried out to estimate the influence of the four TCDs on the secretion of interferon (IFN)-γ by CD8+ T lymphocytes induced by anti-CD3/CD28 antibodies.Statistical analysis was done by nonparametric rank sum test.Results Of the 23 TCDs,14 significantly inhibited the proliferation of CD8+ T lymphocytes (all P ＜ 0.05),of which,Rhizoma Coptidis,Radix Scutellariae,Radix Aucklandiae and Rhizoma Curcumae Longae displayed the strongest inhibitory capacity with the 50％ inhibitory concentration being 25,35,50 and 60 mg/L respectively,and the 100％ inhibitory concentration being 200,100,200 and 200 mg/L respectively.The anti-CD3/CD28 antibody-induced secretion of IFN-γby CD8+ T lymphocytes was markedly suppressed by Radix Scutellariae,Radix Aucklandiae and Rhizoma Curcumae Longae at the concentration of 100 mg/L,but not by Rhizoma Coptidis at this concentration.Conclusions A model for studying the proliferation of CD8+ T lymphocytes is successfully developed in vitro,and four TCDs with strong inhibitory effects on the proliferation of CD8+ T lymphocytes have been screened out with this model. Key words: CD8-positive T-lymphocytes; Drug evaluation, preclinical; Drugs, Chinese herbal; Cell proliferation