Systematic investigation of the material basis, effectiveness and safety of Thesium chinense Turcz. and its preparation Bairui Granules against lung inflammation.

BACKGROUND: Thesium chinense Turcz. (Named as Bai Rui Cao in Chinese) and its preparations (e.g., Bairui Granules) have been used to treat inflammatory diseases, such as acute mastitis, lobar pneumonia, tonsillitis, coronavirus disease 2019 (COVID-19), and upper respiratory tract infection. However, the material basis, pharmacological efficiency, and safety have not been illustrated. METHODS: Anti-inflammatory activity-guided isolation of constituents has been performed using multiple column chromatography, and their structures were elucidated by NMR spectroscopy and ECD calculations. The inhibitory effects on lung inflammation and safety of the crude ethanol extract (CE), Bairui Granules (BG), and the purified active constituents were evaluated using lipopolysaccharide (LPS)-stimulated acute lung inflammation (ALI) mice model or normal mice. RESULTS: Seven new compounds (1-7) and fifty-six known compounds (8-63) were isolated from T. chinense, and fifty-four were reported from this plant for the first time. The new flavonoid glycosides 1-2, new fatty acids 4-5, new alkaloid 7 as well as the known constituents including flavonoid aglycones 8-11, lignans 46-54, alkaloids 34 and 45, coumarins 57, phenylpropionic acids 27, and simple aromatic compounds 39, 44 and 58 exhibited anti-inflammatory activity. Network pharmacology analysis indicated that anti-inflammation of T. chinense was attributed to flavonoids and alkaloids by regulating inflammation-related proteins (e.g., TNF, NF-κB, TGF-ß). Furthermore, constituents of T. chinense including kaempferol-3-O-glucorhamnoside (KN, also named as Bairuisu I, 19), astragalin (AG, Bairuisu II, 12), and kaempferol (KF, Bairuisu III, 8), as well as CE and BG could alleviate lung inflammation caused by LPS in mice by preventing neutrophils infiltration and the expression of the genes for pro-inflammatory cytokines NLRP3, caspase-1, IL-1ß, and COX-2. After a 28-day subacute toxicity test, BG at doses of 4.875 g/kg and 9.750 g/kg (equivalent to onefold and twofold the clinically recommended dose) and CE at a dose of 11.138 g/kg (equivalent to fourfold the clinical dose of BG) were found to be safe and non-toxic. CONCLUSIONS: The discovery of sixty-three constituents comprehensively illustrated the material basis of T. chinense. T. chinense and Bairui Granules could alleviate lung inflammation by regulating inflammation-related proteins and no toxicity was observed under the twofold of clinically used doses.

Integrated network pharmacology and pharmacological investigations to explore the potential mechanism of Ding-Chuan-Tang against chronic obstructive pulmonary disease.

ETHNOPHARMACOLOGICAL RELEVANCE: Ding-Chuan-Tang (Abbreviated as DCT) is frequently prescribed for treatment of respiratory diseases, including chronic obstructive pulmonary disease (COPD), which is characterized by coughing, wheezing, and chest tightness in traditional Chinese medicine (TCM). However, the potential mechanism of DCT has not been investigated. AIM OF STUDY: The aim of the study is to explore the efficiency of DCT in the treatment of COPD in vivo and in vitro, and to illustrate the possible mechanism against COPD. METHODS: COPD model was induced by exposure of mice to cigarette smoke (CS) for 16 weeks. Enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay, Western blot, etc., were used to explore the efficiency and mechanisms of DCT. Network pharmacology analysis, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, etc., was performed to explore the potential targets in the treatment of DCT on COPD. RESULTS: DCT significantly alleviated pulmonary pathological changes in mouse COPD model, and inhibited inflammatory response induced by CS and LPS in vivo and in vitro. Network pharmacology analysis suggested that DCT alleviated COPD via inhibiting inflammation by regulating PI3K-AKT pathway. In cell-based models, DCT suppressed the phosphorylation of PI3K and AKT, which further regulated its downstream targets Nrf2 and NF-κB, and inhibited inflammatory response. CONCLUSIONS: DCT effectively attenuated COPD in the mouse model induced by CS. The therapeutic mechanism of DCT against COPD was closely associated with the regulation of PI3K-AKT pathway and its downstream transcription factors, Nrf2 and NF-κB.

Methotrexate Chemotherapy Causes Growth Impairments, Vitamin D Deficiency, Bone Loss, and Altered Intestinal Metabolism-Effects of Calcitriol Supplementation.

Vitamin D deficiency or insufficiency is prevalent in childhood cancer patients and survivors after chemotherapy; further studies are needed to investigate the underlying aetiology and effectiveness of vitamin D supplementation in preventing chemotherapy-induced bone loss. This study used a rat model of treatment with antimetabolite methotrexate to investigate whether methotrexate chemotherapy causes vitamin D deficiency and if vitamin D supplementation attenuates the resultant bone loss. Methotrexate treatment (five daily injections) decreased serum vitamin D levels (from 52 to <30 ng/mL), reduced body and bone lengthening and tibial trabecular bone volume, and altered intestinal vitamin D metabolism, which was associated with intestinal mucosal damage known to cause malabsorption of nutrients, including dietary vitamin D and calcium. During the early stage after chemotherapy, mRNA expression increased for vitamin D activation enzyme CYP27B1 and for calcium-binding protein TRPV6 in the intestine. During the intestinal healing stage, expression of vitamin D catabolism enzyme CYP24 increased, and that of TRPV6 was normalised. Furthermore, subcutaneous calcitriol supplementation diminished methotrexate-induced bone loss due to its effect suppressing methotrexate-induced increased bone resorption. Thus, in young rats, methotrexate chemotherapy causes vitamin D deficiency, growth impairments, bone loss, and altered intestinal vitamin D metabolism, which are associated with intestinal damage, and vitamin D supplementation inhibits methotrexate-induced bone loss.

Chemotherapy-Induced Intestinal Microbiota Dysbiosis Impairs Mucosal Homeostasis by Modulating Toll-like Receptor Signaling Pathways.

Chemotherapy-induced intestinal mucositis, a painful debilitating condition affecting up to 40-100% of patients undergoing chemotherapy, can reduce the patients' quality of life, add health care costs and even postpone cancer treatment. In recent years, the relationships between intestinal microbiota dysbiosis and mucositis have drawn much attention in mucositis research. Chemotherapy can shape intestinal microbiota, which, in turn, can aggravate the mucositis through toll-like receptor (TLR) signaling pathways, leading to an increased expression of inflammatory mediators and elevated epithelial cell apoptosis but decreased epithelial cell differentiation and mucosal regeneration. This review summarizes relevant studies related to the relationships of mucositis with chemotherapy regimens, microbiota, TLRs, inflammatory mediators, and intestinal homeostasis, aiming to explore how gut microbiota affects the pathogenesis of mucositis and provides potential new strategies for mucositis alleviation and treatment and development of new therapies.

Retracted: Icariin attenuates methotrexate chemotherapy-induced bone marrow microvascular damage and bone loss in rats.

Methotrexate (MTX), a widely used antimetabolite in paediatric cancer to treatment, has been widely reported to cause bone loss and bone marrow (BM) microvascular (particularly sinusoids) damage. Investigations must now investigate how MTX-induced bone loss and microvasculature damage can be attenuated/prevented. In the present study, we examined the potency of icariin, an herbal flavonoid, in reducing bone loss and the dilation/damage of BM sinusoids in rats caused by MTX treatment. Groups of young rats were treated with five daily MTX injections (0.75 mg/kg) with and without icariin oral supplementation until Day 9 after the first MTX injection. Histological analyses showed a significant reduction in the bone volume/tissue volume (BV/TV) fraction (%) and trabecular number in the metaphysis trabecular bone of MTX-treated rats, but no significant changes in trabecular thickness and trabecular spacing. However, the BV/TV (%) and trabecular number were found to be significantly higher in MTX + icariin-treated rats than those of MTX alone-treated rats. Gene expression analyses showed that icariin treatment maintained expression of osteogenesis-related genes but suppressed the induction of adipogenesis-related genes in bones of MTX-treated rats. In addition, icariin treatment attenuated MTX-induced dilation of BM sinusoids and upregulated expression of endothelial cell marker CD31 in the metaphysis bone of icariin + MTX-treated rats. Furthermore, in vitro studies suggest that icariin treatment can potentially enhance the survival of cultured rat sinusoidal endothelial cells against cytotoxic effect of MTX and promote their migration and tube formation abilities, which is associated with enhanced production of nitric oxide.

Chinese herbal medicines in the prevention and treatment of chemotherapy-induced nausea and vomiting.

PURPOSE OF REVIEW: Chemotherapy-induced nausea and vomiting (CINV) critically impact on cancer patients' quality of life, causing various complications, and increasing the risk of chemotherapeutic failure. Current treatments addressing CINV are reasonably costly and have many side effects. Chinese herbal medicines (CHMs) have been widely used in treating CINV in China. Clinical trials and modern experimental studies on CINV using CHMs are reviewed for clues that may help us to better prevent and treat CINV with CHMs, and develop safe and effective treatments. RECENT FINDINGS: Since 1985, 92 clinical trials on CINV were identified, which comprised of 3778 patients with different cancer types, chemotherapy regimens, prescriptions, treatment durations, and combinations with Western medicines. The most frequently used herbs are Pinellia Rhizoma, Glycyrrhizae Radix et Rhizoma, Poria, and Atractylodis Macrocephalae Rhizoma, aiming to regulate the flow of Qi and remove phlegm and dampness residing in the stomach and spleen. Although all of the reported trials are randomized open trials, the curative effects range from 55.81 to 100%. SUMMARY: CHMs seem effective in the prevention and treatment of CINV based on the clinical trials and some animal experiments, although the scientific validity of many clinical studies remain to be confirmed as many of them were inadequately designed. Further well designed clinical trials and more preclinical studies with reliable markers are required to confirm the usefulness of CHMs as complementary medicines for treating CINV and to use CHMs as rational prescriptions rather than based only on empirical knowledge.

Identification of novel Nrf2 activators from Cinnamomum chartophyllum H.W. Li and their potential application of preventing oxidative insults in human lung epithelial cells.

Human lung tissue, directly exposed to the environmental oxidants and toxicants, is apt to be harmed to bring about acute or chronic oxidative insults. The nuclear factor erythroid 2-related factor 2 (Nrf2) represents a central cellular defense mechanism, and is a target for developing agents against oxidative insult-induced human lung diseases. Our previous study found that the EtOH extract of Cinnamomum chartophyllum protected human bronchial epithelial cells against oxidative insults via Nrf2 activation. In this study, a systemic phytochemical investigation of the aerial parts of C. chartophyllum led to the isolation of thirty chemical constituents, which were further evaluated for their Nrf2 inducing potential using NAD(P)H: quinone reductase (QR) assay. Among these purified constituents, a sesquiterpenoid bearing α, ß-unsaturated ketone group, 3S-(+)-9-oxonerolidol (NLD), and a diphenyl sharing phenolic groups, 3, 3', 4, 4'-tetrahydroxydiphenyl (THD) significantly activated Nrf2 and its downstream genes, NAD(P)H quinone oxidoreductase 1 (NQO-1), and γ-glutamyl cysteine synthetase (γ-GCS), and enhanced the nuclear translocation and stabilization of Nrf2 in human lung epithelial cells. Importantly, NLD and THD had no toxicities under the Nrf2 inducing doses. THD also demonstrated a potential of interrupting Nrf2-Keap1 protein-protein interaction (PPI). Furthermore, NLD and THD protected human lung epithelial cells against sodium arsenite [As(III)]-induced cytotoxicity. Taken together, we conclude that NLD and THD are two novel Nrf2 activators with potential application of preventing acute and chronic oxidative insults in human lung tissue.

Screening of traditional Chinese medicines with therapeutic potential on chronic obstructive pulmonary disease through inhibiting oxidative stress and inflammatory response.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a major public health problem and gives arise to severe chronic morbidity and mortality in the world. Inflammatory response and oxidative stress play dominant roles in the pathological mechanism of COPD, and have been regarded to be two important targets for the COPD therapy. Traditional Chinese medicines (TCMs) possess satisfying curative effects on COPD under guidance of the TCM theory in China, and merit in-depth investigations as a resource of lead compounds. METHODS: One hundred ninety-six of TCMs were collected, and extracted to establish a TCM extract library, and then further evaluated for their potency on inhibitions of oxidative stress and inflammatory response using NADP(H):quinone oxidoreductase (QR) assay and nitric oxide (NO) production assay, respectively. RESULTS: Our investigation observed that 38 of the tested TCM extracts induced QR activity in hepa 1c1c7 murine hepatoma cells, and 55 of them inhibited NO production in RAW 264.7 murine macrophages at the tested concentrations. Noteworthily, 20 of TCM extracts simultaneously inhibited oxidative stress and inflammatory responses. CONCLUSION: The observed bioactive TCMs, particularly these 20 TCMs with dual inhibitory effects, might be useful for the treatment of COPD. More importantly, the results of the present research afford us an opportunity to discover new lead molecules as COPD therapeutic agents from these active TCMs.

Steamed root of Rehmannia glutinosa Libosch (Plantaginaceae) alleviates methotrexate-induced intestinal mucositis in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Intestinal mucositis induced by chemotherapy is a severe clinical problem in cancer patients that currently lack effective interventions. In traditional Chinese medicine, chemotherapeutic toxicity is diagnosed as Qi and Yin deficiency, and steamed rehmannia root (SRR) is frequently prescribed to these patients. Whether SRR can prevent the adverse effects remains to be confirmed experimentally. The present study used a rat model to investigate potential efficacy and action mechanisms of SRR in attenuating the adverse effects caused by chemotherapy. MATERIALS AND METHODS: Intraperitoneal injection of a single dose of anti-metabolite methotrexate (MTX, 25mg/kg) was given to adult Wistar rats, which also received oral gavage of water or SRR (1.08g/kg twice daily 3 days before and 4 days after MTX treatment), or calcium folinate (CF, a clinically used MTX antidote as a comparison, at 1mg/kg twice daily 36h after MTX treatment), or SRR and CF in combination. Animals were sacrificed 4 days after MTX treatment. Complete blood cell counting was carried out. Jejunum was analyzed histologically for mucosal damage, immunohistochemically for proliferating cell nuclear antigen (PCNA), and biochemically for thiobarbituric acid reactive substances (TBARS) and reduced glutathione (GSH), as well as for tumor necrosis factor alpha (TNF-α). RESULTS: MTX treatment led to weight loss, leucopenia, polycythemia, increase in large thrombocyte ratio, intestinal villus atrophy, crypt loss and reduction in PCNA positive crypt cells, increases in mucosal TBARS and TNF-α and decrease in GSH. All these alterations were inhibited by SRR administration except leucopenia, and the effects of CF or CF plus SRR supplementation were found to be inferior to those of SRR. CONCLUSIONS: SRR can alleviate MTX-induced gut mucositis, which may be achieved by inhibiting MTX-induced oxidative stress and inflammatory response. These findings support the application of SRR in chemotherapy but not the combined application of SRR and CF.

Characteristics and kinetics of catalpol degradation and the effect of its degradation products on free radical scavenging.

BACKGROUND: The dried and steamed roots of Rehmannia glutinosa have different pharmacological functions and indications. Catalpol, the main active component of the dried root, was found to be entirely degraded together with amino acids and some oligosaccharides during preparation of the steamed root. Its degradation may contribute to the differences between dried and steamed roots. OBJECTIVE: To reveal the characteristics and kinetics of catalpol degradation, and evaluate its influence on the antioxidant properties of steamed Rehmannia roots. MATERIALS AND METHODS: Purified catalpol was heated under different pH and temperature values for different times, alone or with sugars or amino acids. Catalpol concentration was determined by high-performance liquid chromatography. Browning was expressed by the absorbance at 420 nm (A420), and antioxidation was displayed by 2,2-diphenyl-1-picrylhydrazyl free radical scavenging ability (SADPPH). Activation energy was calculated using Arrhenius plotting. RESULTS: Catalpol was stable in neutral conditions and sensitive to acidic pH under high temperatures. Sugars had no influence on catalpol degradation; however, most amino acids, except for proline, could promote the degradation, and were associated with an increase in A420 and SADPPH values. These changes were proved to be mainly related with catalpol aglycone and were dependent on the presence of amino acids. Catalpol degradation was found to obey first-order kinetics. The activation energies were 81.7, 88.8 and 98.7 kJ/mol at pH 4.0, 5.0, and 6.0 respectively, and 70.7 kJ/mol at pH 4.0 value and in the presence of glycine. CONCLUSIONS: Catalpol degradation, especially, in the presence of amino acids can substantially boost antioxidant properties of the products; therefore, the traditional method for processing Rehmannia root seems rather apt.

[Discussion on the quality marker of rehmanniae radix praeparata in the Chinese Pharmacopoeia].

OBJECTIVE: In the Chinese Pharmacopoeia (2010 edition), the minimum limit of verbascoside (acteoside) in Rehmanniae Radix Praeparata (RRP) was set at 0.20%, the rationality of the standard was evaluated in this paper. METHODS: 10 samples of Rehmanniae Radix (RR) and RRP were collected or prepared. According to the traditional method, different samples of RRP were prepared from the same batch of RR, and the content of verbascoside was determined by HPLC. RESULTS: The average content of verbascoside in RR and RRP was 0.0226% and 0.0097%, respectively, and their difference was up to the level of P < 0.01. The content of verbascoside was found decreased with processing time increasing. Long-time storage also results in substantial loss of verbascoside. CONCLUSION: Processing results in verbascoside decreasing significantly. It is unreasonable that RR and RRP have the same standard in the content of verbascoside. It is suggested that the maximum limit of verboscoside should be set in the new version of Chinese pharmacopoeia.

Yeast exposure in the preparation of steamed rehmannia root improving its effects on alloxan-induced diabetic rats.

ETHNOPHARMACOLOGICAL RELEVANCE: The root of Rehmannia glutinosa (Gaertn.) DC. has been used in China for more than 2000 years to treat various diseases including diabetes mellitus (DM) and its complications. Steamed rehmannia root (SRR) is one of the mainly used forms. During its preparation, the material has the chance to be exposed to microorganisms, particularly yeast. The aim of this study is to verify the effect of SRR on diabetes complications and the necessity of yeast exposure. MATERIALS AND METHODS: Water extract of SRR was incubated with alcohol yeast to obtain fermented SRR (FSRR). Alloxan-induced diabetic rats were administrated with medicated animal chows for 8 weeks. Urine volume, fasted blood glucose and food intake were monitored, and open field test and tail immersion test were conducted in the last week, plasma and urine samples were subjected to biochemical examinations. RESULTS: In DM rats, defecation in open field test was found reduced, and tail flick latency in tail immersion test increased. In the meantime, urinary excretions of Na(+), K(+), aldosterone, albumin and creatinine increased, and plasma concentrations of Na(+), K(+) and creatinine reduced and those of aldosterone, TXB2/6-Keto-PGF(1α) and urea nitrogen elevated. Most of these indicators were significantly improved by FSRR administration, but the effects of SRR were relatively inferior in several aspects. However, SRR and FSRR could not improve the typical symptoms of DM. CONCLUSIONS: Our data demonstrated that both SRR and FSRR have no obvious hypoglycemic effect, but have the potential to prevent the onset and development of diabetes complications, and this function can be improved by yeast exposure.

[Simultaneous determination of 6 active components in Chrysanthemum morifolium by HPLC].

OBJECTIVE: To develop a HPLC method quantitative method for simultaneous determination of chlorogenic acid, 1, 5-dicaffeoylquinic acid, isochlorogenic acid A, isochlorogenic acid C, luteolin-7-O-beta-D-glucoside and apigenin-7-O-beta-D-glucoside in Chrysanthemum morifolium Ramat. METHOD: A Phenomenex Gemini-NX C18 column (4.6 mm x 250 mm, 5 microm) was used with CH3 OH and 0.4% H3PO4 as mobile phases. The flow rate was 1 mL x min(-1), the column temperature was 25 degrees C, and the detection wavelength was set at 350 nm. RESULT: The 6 active components were in baseline separation. The linearity of this method was good (r > or = 0.999 7), and the average recoveries were 100.6% - 102.4%, RSD < 3%. Except isochlorogenic acid A, the contents of the determined components in the steam-blanched flower heads were significantly higher than those non blanched. The contents of chlorogenic acid and isochlorogenic acid A in the steam-blanched semiopened flower heads were higher than fully opened ones by 53% and 41%, respectively. CONCLUSION: The method is sensitive, accurate, reliable and repeatable, which can be used for quality evaluation of Chrysanthemum.

Antidepressant effect of Shudihuang on mice exposed to unpredictable chronic mild stress.

AIM OF THE STUDY: Depression is a severe mood disorder. It was treated with Shudihuang, the steamed roots of Rehmannia glutinota Libosch. (SRG), in traditional Chinese medicine. The present paper was designed to verify its antidepressant effect. MATERIALS AND METHODS: A mouse model of depression was established though unpredictable chronic mild stress (UCMS). Low and high doses of SRG were administered orally. Fur state, body and organ weight, and gastric ulcers were examined. Locomotion was assayed in open field test. Liver antioxidant indexes were measured spectrophotometrically. RESULTS: Fur state, body and organ weight were found to be insensitive to UCMS. The locomotion reduced by UCMS was restored by low dose of SRG (2.5 g/kg BW) but not by high dose (5 g/kg BW). UCMS resulted in aggravated gastric ulceration, elevated liver malondialdehyde, together with reduced total antioxidant capability, glutathione content, and superoxide dismutase and catalase activities. The alterations were improved by SRG in a dose-dependent manner. The differences in the activity of glutathione peroxidase were statistically nonsignificant among groups. Clomipramine the positive drug was similar to SRG especially in antioxidation. CONCLUSION: SRG is of therapeutic value for depression-like disorders, and antioxidation may be one of the mechanisms underlying its antidepressant action.

[Optimization of manninotriose preparation from stachoyse].

OBJECTIVE: To optimize the processing of mannatriose preparation from stachyose by acidolysis. METHODS: An orthogonal experiment was performed to select a better condition of acidolysis. Fructoes and stachyose was determined by HPLC. RESULTS: Among the factors set in the present work, temperature and time have little influence on the acidolysis, while the effect of pH value was significant (P < 0.05). The best processing was selected as heating stachosy solution for 12h at pH 2.5 and 90 degrees C. CONCLUSION: It is practicable to prepare manninotriose from stachyoses by acidolysis.

[Advances in research on pharmacodynamics and chemical conversion of catalpol].

Catalpol is a active component in Rehmannia Root, and its pharmacological effects are extensive. In this paper its effects and chemical changes were summarized. This will offer the reference for further research work catalpol.

[Extraction and salting-out purification of alpha-galactosidase and beta-glucosidase from fresh roots of Rehmannia glutinosa].

OBJECTIVE: To extract and preliminarily purify alpha-galactosidase and beta-glucosidase from the fresh roots of Rehmannia glutinosa. METHODS: With the enzyme activity as a criterion, the best procedure of extraction was selected though orthogonal design method, and the desired saturation of ammonium sulfate in two-step salting-out was settled by gradient sedimentation of root extract according to enzyme activity and protein content. RESULTS: Temperature and solvent volume affect the extraction of alpha-galactosidase significantly, while solvent type to beta-glucosidase. Therefore the procedure for extracting two enzymes was decided as mixing comminuted fresh root with 3 times phosphate buffer, and placing the mixture in refrigeratory at 4 degrees C for 4 hours, and then obtaining the enzyme liquid by centrifuging at 4 degrees C. 30% and 60% saturation was defined as the lower and upper point for two-step ammonium sulfate salting-out of the two enzymes. CONCLUSION: alpha-Galactosidase and beta-glucosidase exist in the fresh roots, and can be preliminarily purified through two-step salting-out.

[Advances in study on chemical constituents from plants of genus Pfaffia and their bioactivities].

The progress in the studies on chemical constituents and pharmacological activity of the genus Pfaffia is summarized in recent 20 years. These plants contain various chemical constituents and have broad bioactivities such as sthenic, anti-tumor, analgesic and anti-inflammatory and should be further investigated.

[Preservation of germplasm of Rehmannia glutinosa in vitro].

OBJECTIVE: To establish a method for the germplasm preservation of R. glutinosa. METHOD: Plantlets of different cultivars obtained by tip culture were inoculated into test tubes with MS medium supplemented with 0.2 mg.L-1 BA and 0.02 mg.L-1 NAA, and preserved at 4-6 degrees C in dark. At the same time, different media (A. distilled water + 10 g.L-1 agar; B. 1/2 MS + 5 g.L-1 agar; C. MS + 10 g.L-1 agar; D. 1/2 MS + 0.5 BA + 0.02 NAA + 10 g.L-1 agar; E. MS + 0.5 BA + 0.02 NAA + 10 g.L-1 agar) were set to conserve plantlets of "85-5" on the same condition. RESULT: 4 months later, the death rate of "Xinggeda" was 73%, "Tucheng" 60%, "85-5" 33%, and "Beijing 1" 9%. All of the plantlets of "85-5" in different media kept alive. 10 months later, most of the preserved plantlets browned and wilted except those on medium A. CONCLUSION: The low temperature endurance of R. glutinosa is cultivar-dependent. Medium A can preserve "85-5" for more than 10 months in vitro.

[Photosynthetic pigment contents of different germplasm of Rehmannia glutinosa and the relationship between pigments and leaf color].

OBJECTIVE: To evaluate the germplasm of Rehmannia glutinosa on the basis of photosynthetic pigment contents (PPC). METHOD: 20 cultivars were planted on the same condition. On Oct. 23 and Sept. 25, 3 leaves per cultivar were collected on different plants, and 80 mg mesophyll was collected among upper lateral veins and was ground in 96% alcohol, and the supernatant was subjected to measure on a spectrophotometer (Angilent 8453). RESULT: The PPCs among cultivars were significantly different at a P < or = 0.01 level. The results of the measurements were similar. Chlolophyll a was the most abundant pigment, but varied to a great extent among different cultivars. 20 cultivars were divided into 9 homogeneous groups according to the contents of chlorophyll a by Duncan's multiple range test at P < or = 0.05. In addition, the content of chlorophyll a was closely related to leaf color. The cultivars with higher chlolophyll a had deep green leaves, and those with lower had yellow green or pale green leaves. CONCLUSION: PPC was an inherent character and an important index for the germplasm evaluation of R. glutinosa.