ThermomiR-377-3p-induced suppression of Cirbp expression is required for effective elimination of cancer cells and cancer stem-like cells by hyperthermia.

BACKGROUND: In recent years, the development of adjunctive therapeutic hyperthermia for cancer therapy has received considerable attention. However, the mechanisms underlying hyperthermia resistance are still poorly understood. In this study, we investigated the roles of cold­inducible RNA binding protein (Cirbp) in regulating hyperthermia resistance and underlying mechanisms in nasopharyngeal carcinoma (NPC). METHODS: CCK-8 assay, colony formation assay, tumor sphere formation assay, qRT-PCR, Western blot were employed to examine the effects of hyperthermia (HT), HT + oridonin(Ori) or HT + radiotherapy (RT) on the proliferation and stemness of NPC cells. RNA sequencing was applied to gain differentially expressed genes upon hyperthermia. Gain-of-function and loss-of-function experiments were used to evaluate the effects of RNAi-mediated Cirbp silencing or Cirbp overexpression on the sensitivity or resistance of NPC cells and cancer stem-like cells to hyperthermia by CCK-8 assay, colony formation assay, tumorsphere formation assay and apoptosis assay, and in subcutaneous xenograft animal model. miRNA transient transfection and luciferase reporter assay were used to demonstrate that Cirbp is a direct target of miR-377-3p. The phosphorylation levels of key members in ATM-Chk2 and ATR-Chk1 pathways were detected by Western blot. RESULTS: Our results firstly revealed that hyperthermia significantly attenuated the stemness of NPC cells, while combination treatment of hyperthermia and oridonin dramatically increased the killing effect on NPC cells and cancer stem cell (CSC)­like population. Moreover, hyperthermia substantially improved the sensitivity of radiation­resistant NPC cells and CSC­like cells to radiotherapy. Hyperthermia noticeably suppressed Cirbp expression in NPC cells and xenograft tumor tissues. Furthermore, Cirbp inhibition remarkably boosted anti­tumor­killing activity of hyperthermia against NPC cells and CSC­like cells, whereas ectopic expression of Cirbp compromised tumor­killing effect of hyperthermia on these cells, indicating that Cirbp overexpression induces hyperthermia resistance. ThermomiR-377-3p improved the sensitivity of NPC cells and CSC­like cells to hyperthermia in vitro by directly suppressing Cirbp expression. More importantly, our results displayed the significantly boosted sensitization of tumor xenografts to hyperthermia by Cirbp silencing in vivo, but ectopic expression of Cirbp almost completely counteracted hyperthermia-mediated tumor cell-killing effect against tumor xenografts in vivo. Mechanistically, Cirbp silencing-induced inhibition of DNA damage repair by inactivating ATM-Chk2 and ATR-Chk1 pathways, decrease in stemness and increase in cell death contributed to hyperthermic sensitization; conversely, Cirbp overexpression-induced promotion of DNA damage repair, increase in stemness and decrease in cell apoptosis contributed to hyperthermia resistance. CONCLUSION: Taken together, these findings reveal a previously unrecognized role for Cirbp in positively regulating hyperthermia resistance and suggest that thermomiR-377-3p and its target gene Cirbp represent promising targets for therapeutic hyperthermia.

Effect of a Traction Exercise Neck Brace on Cervical Spondylopathy Radiculopathy: A Clinical Study and Finite Element Analysis.

Traction of cervical spine is an effective method for the treatment of cervical spondylotic radiculopathy (CSR). In this study, a cervical tractor named traction exercise neck brace (TENB) was used to evaluate its effect on the patients with CSR. Forty CSR volunteers were recruited and randomly divided into two groups. One group was subjected to cervical muscle exercise with TENB under static traction condition. Another group was subjected to (JOBT) as controls. Symptoms of CSR were evaluated by the visual analogue scale (VAS) and neck disability index (NDI). Imaging characteristics were assessed by curvature of the cervical spine and size of the intervertebral foramen. A finite element (FE) analysis model of cervical spine was established by 3D reconstruction to simulate the TENB traction, which evaluates the biomechanical performance. Results showed that TENB significantly reduced scores of VAS and NDI in subjects, and this improved effect on symptoms of pain and radiculopathy is better than that of JOBT. TENB also improved the cervical curvature and enlarged intervertebral foramen at the C4-C6 level. Moreover, FE analysis found that simulated TENB traction increased the spacing of intervertebral foramen, intervertebral disc, and zygapophyseal and uncovertebral joints and changed the stress distribution on the facet joints and nucleus pulposus. This study demonstrates that TENB relieves the symptoms of CSR by adjusting structure of cervical vertebra and restoring its biomechanical performance, which may be a promising instrument in the treatment of CSR.

Rapid quantification of multi-components in alcohol precipitation liquid of Codonopsis Radix using near infrared spectroscopy (NIRS).

A near infrared spectroscopy (NIRS) approach was established for quality control of the alcohol precipitation liquid in the manufacture of Codonopsis Radix. By applying NIRS with multivariate analysis, it was possible to build variation into the calibration sample set, and the Plackett-Burman design, Box-Behnken design, and a concentrating-diluting method were used to obtain the sample set covered with sufficient fluctuation of process parameters and extended concentration information. NIR data were calibrated to predict the four quality indicators using partial least squares regression (PLSR). In the four calibration models, the root mean squares errors of prediction (RMSEPs) were 1.22 µg/ml, 10.5 µg/ml, 1.43 µg/ml, and 0.433% for lobetyolin, total flavonoids, pigments, and total solid contents, respectively. The results indicated that multi-components quantification of the alcohol precipitation liquid of Codonopsis Radix could be achieved with an NIRS-based method, which offers a useful tool for real-time release testing (RTRT) of intermediates in the manufacture of Codonopsis Radix.

Comprehensive Quantitative Analysis of SQ Injection Using Multiple Chromatographic Technologies.

Quality control of Chinese medicine injections remains a challenge due to our poor knowledge of their complex chemical profile. This study aims to investigate the chemical composition of one of the best-selling injections, Shenqi Fuzheng (SQ) injection (SQI), via a full component quantitative analysis. A total of 15 representative small molecular components of SQI were simultaneously determined using ultra-high performance liquid chromatography (UHPLC) coupled with quadrupole tandem time-of-flight mass spectrometry (Q-TOF-MS); saccharide composition of SQI was also quantitatively determined by high performance liquid chromatography (HPLC) with evaporative light scattering detector (ELSD) on an amino column before and after acid hydrolysis. The existence of polysaccharides was also examined on a gel permeation chromatography column. The method was well validated in terms of linearity, sensitivity, precision, accuracy and stability, and was successfully applied to analyze 13 SQI samples. The results demonstrate that up to 94.69% (w/w) of this injection product are quantitatively determined, in which small molecules and monosaccharide/sucrose account for 0.18%-0.21%, and 53.49%-58.2%, respectively. The quantitative information contributes to accumulating scientific evidence to better understand the therapy efficacy and safety of complex Chinese medicine injections.

In vitro assays suggest Shenqi Fuzheng Injection has the potential to alter melanoma immune microenvironment.

ETHNOPHARMACOLOGICAL RELEVANCE: A modern agent Shenqi Fuzheng Injection (SFI), prepared from Codonopsis Radix and Astragali Radix, has been commonly used as a supplementary therapy for cancers including melanoma. This agent was derived from a formula documented in the "National Collection of Chinese Medicine Prescriptions". The formula has long been used as a remedy for Qi deficiency that is closely associated with cancer-related fatigue and poor quality of life. However, the antimelanoma mechanisms of SFI remain unclear. Here we tested if SFI exerted antimelanoma effects by reprograming the tumour immune microenvironment using in vitro assays. MATERIALS AND METHODS: The cytotoxic activities of Jurkat T cells when co-cultured with A375 cells were determined in the presence or absence of SFI. The migratory activities of Jurkat T cells were examined in the transwell assay system. The mRNA expression and production of cytokines (IL-10, TGF ß and VEGF) in A375 cells in the presence or absence of SFI were determined by real-time PCR and ELISA, respectively. RESULTS: When A375 cells were co-cultured with Jurkat T cells in the presence of SFI (220µg/mL), a potent cytotoxicity effect against A375 cells was observed. Supernatants from A375 cells that were treated with SFI (110 and 220µg/mL) significantly increased the migratory capacity of Jurkat T cells in transwell assays. SFI also markedly reduced the mRNA expression levels and the release of immunosuppressive cytokines IL-10, TGF-ß and VEGF in A375 cells in a concentration-dependent manner. CONCLUSIONS: SFI enhanced the cytotoxic and migratory activities of Jurkat T cells towards A375 melanoma cells. The effects were associated with SFI's suppression on immunosuppressive cytokines for their release from and gene expressions in A375 melanoma cells. These in vitro findings suggested that SFI might reprogramme the immunosuppressive melanoma microenvironment in vivo to enhance the cytotoxicity of tumour-infiltrating immune cells. This study provides a pharmacological basis for the adjunctive use of SFI in melanoma treatment.

[Simultaneous determination of six components in water extract and alcohol precipitation liquid in Astragali Radix by HPLC-UV-ELSD].

A HPLC-UV-ELSD method was established for simultaneous determination of six components in two intermediates of Shenqi Fuzheng injection (SFI) and the feasibility of establishing quantitative analysis of multi-components by single marker (QAMS) methods on different detectors was further explored. Calycosin-7-O-ß-D-glucoside and astragloside Ⅳ were selected as internal reference substances for respectively flavonoids and saponins, and relative correlation factors (RCF) of formononetin-7-O-ß-D-glucoside, 9, 10-dimethoxypterocarpan-3-O-ß-D-glucopyranoside, 2'-dihydroxy-3', 4'-dimethoxyisoflavan-7-O-ß-D-glucopyranoside and astragloside Ⅱ were calculated. Eventually, quantitative results of the 14 samples were compared between QAMS and external standard method. The sample concentrations calculated by QAMS were similar with concentrations calculated by external standard method, and the absolute values of relative deviations were generally less than 5% according to the UV detection of flavonoids. On the basis of ELSD detection for saponins, however, the absolute values of relative deviation of the two methods ranged from 0.48% to 23.17%. The QAMS method built on ultraviolet (UV) detectors was stable and can be used as a substitute method to reduce the consumption of standard compounds; meanwhile, the accuracy of QAMS method built on evaporative light scattering detector (ELSD) was inferior to that of external standard method, and the working principle of ELSD and feasible concentration range remain to be further studied.

[A new bisabolane-type sesquiterpenoid from Coreopsis tinctoria].

To study the chemical constituents of the inflorescences of Coreopsis tinctoria from Xinjiang, isolation and purification of constituents were carried out by column chromatography on macroporous resin (D101) , MCI gel, MDS gel, silica gel, Sephadex LH-20 and semi-preparative HPLC. The structures of the compounds were identified by physicchemical properties and spectral data analysis. Fourteen compounds were isolated and identified as coretinterpenoid A (1), coretinphenol (2), quercetin (3), quercetin-3-O-ß-glucopyranoside (4), luteolin (5), taxifolin (6), 7, 3', 5'-trihydroxyflavanone (7), isookanin (8), isookanin-7-O-ß-D-glucopyranoside (9), 5, 7, 3', 5'-tetrahydroxyflavanone-7-O-ß-D-glucopyranoside (10), butein (11), okanin (12), sulfuretin (13), and linocinnamarin (14). Compound 1 was a new isabolane-type sesquiterpenoid and compounds 4, 10 and 13 were isolated from this plant for the first time.

[Detection of agent "zhuanggenling" and investigation of utilization of plant growth retardants in traditional Chinese medicine cultivation].

Plant growth retardant as one of plant growth regulator can inhibit the cell division, elongation and growth rate in shoot apical meristem (SAM), which can be reversed by gibberellin regulate the product of photosynthesis transfer to the root and rhizome part. As commonly used plant growth retardant, paclobutrazol, uniconazole, chlorocholine chloride, mepiquat chloride, choline chloride and daminozide are used to promote the growth of root and rhizome, call as "zhuanggenling", "pengdasu", "pengdaji" etc. Single or recombination of plant growth regulator is registered as pesticide, and called as pesticide "zhuanggenling" in this paper. Growth regulator which registered as a foliar fertilizer or fertilization was called agricultural fertilizer "zhuanggenling" in this paper. The author investigate the usage of "zhuanggenling" in the root and rhizome of medicinal plants cultivation from 2012 to 2014 in Sichuan province, Huangyuan town, Mianyang (Ophiopogonis Radix); Pengzhou Aoping town (Chuanxiong Rhizoma); Pengshan Xiejia town (Alismatis Rhizoma); Jiangyou Taiping town and Zhangming town (Aconiti Lateralis Radix Praeparata); Yunnan Wenshan (Notoginseng Radix et Rhizoma); Henan province, Wuzhidafeng Town (Rehmanniae Radix, Achyranthis Bidentatae Radix, Dioscoreae Rhizoma); Gansu Min county (Codonopsis Radix, Angelicae Sinensis Radix); Gansu Li county (Rhei Radix et Rhizoma). The result showed that "zhuanggenling" were applied in the most medicinal plant cultivation except Rhei Radix et Rhizoma. It has been applied widespreadly in Ophiopogonis Radix, Alismatis Rhizoma, Achyranthis Bidentatae Radix, Codonopsis Radix; Rehmanniae Radix, commonly in Angelicae Sinensis Radix application, and occasionally in Chuanxiong Rhizoma, Aconiti Lateralis Radix Praeparata, Notoginseng Radix et Rhizoma and Dioscoreae Rhizoma. In 53 collected sample from plantation areas, fifteen (28%) were pesticide "zhuanggenling", thirty-eight (72%) were pesticide "zhuanggenling". UPLC analysis results showed that 38 farmers fertilizer "zhuanggenling" content of 6 kinds of plant growth retardant. It is regarded that fertilizer "zhuanggenling" was dominant in medicinal plant cultivation, and that the plant growth retardant is added widespreadly in farm fertilizer "zhuanggenling". All evidence proves conclusively that "zhuanggenling" have been used in the proper way, whereas some others have been misused or even abused in the use regarding to type, number, use frequency. The root or rhizoma are increased to 20%-200%. But there is lack of evaluation to appraise the quality of medicinal materials from the aspects of research or industry. "zhuanggenling" has become a important Chemical control material besides fertilizer, insecticidal sterilization of pesticide

[Design space approach to optimize first ethanol precipitation process of Dangshen].

Design space approach is applied in this study to enhance the robustness of first ethanol precipitation process of Codonopsis Radix (Dangshen) by optimizing parameters. Total flavonoid recovery, dry matter removal, and pigment removal were defined as the process critical quality attributes (CQAs). Plackett-Burman designed experiments were carried out to find the critical process parameters (CPPs). Dry matter content of concentrated extract (DMCE), mass ratio of ethanol to concentrated extract (E/C ratio) and concentration of ethanol (CEA) were identified as the CPPs. Box-Behnken designed experiments were performed to establish the quantitative models between CPPs and CQAs. Probability based design space was obtained and verified using Monte-Carlo simulation method. According to the verification results, the robustness of first ethanol precipitation process of Dangshen can be guaranteed by operating within the design space parameters. Recommended normal operation space are as follows: dry matter content of concentrated extract of 45.0% - 48.0%, E/C ratio of 2.48-2.80 g x g(-1), and the concentration of ethanol of 92.0% - 92.7%.

A new phenylpropanoid and a new isoflavone glycoside from Shenqi Fuzheng Injection.

A new phenylpropanoid and a new isoflavone glycoside were isolated from Shenqi Fuzheng Injection. Their structures were elucidated as (αS)-α-ethenyl-4-hydroxy-3-methoxy-benzenemethanol (1) and calycosin 7-O-[α-d-glucopyranosyl (1 â†’ 4)]-ß-d-glucopyranoside (2) by means of spectroscopic methods including UV, IR, HR-ESI-MS, and NMR. The absolute configurations of 1 and 2 were confirmed by quantum chemical calculation and acid hydrolysis.

Anticancer effect of total annonaceous acetogenins on hepatocarcinoma.

OBJECTIVE: To confirm the anticancer effect of total annonaceous acetogenins (TAAs) abstracted from Annona squamosa Linn. on human hepatocarcinoma. METHODS: The inhibitory effect of TAAs was demonstrated in H22-bearing mice. The potency of TAAs was confirmed as its 50% inhibiting concentration (IC50) on Bel-7402 cell under Sulfur Rhodamine B staining. Both underlying mechanisms were explored as cellular apoptosis and cell cycle under flow cytometry. Mitochondrial and recipient apoptotic pathways were differentiated as mitochondrial membrane potential under flow cytometry and caspases activities under fluorescence analysis. RESULTS: The inhibitory rate of TAAs in mice was 50.98% at 4 mg/kg dose. The IC50 of TAAs on Bel-7402 was 20.06 µg/mL (15.13-26.61µg/mL). Effective mechanisms of TAAs were confirmed as both of arresting cell cycle at G1 phase and inducing apoptosis dose- and time-dependently. Mitochondrial and recipient pathways involved in apoptotic actions of TAAs. CONCLUSION: TAAs is effective for hepatocarcinoma, via inhibiting proliferation and inducing apoptosis.

Two new acylated flavonol glycosides from the seeds of Lepidium sativum.

Two new acylated flavonol glycosides named kaempferol-3-O-(2-O-sinapoyl)-ß-D-galactopyranosyl-(1 → 2)-ß-D-glucopyranoside-7-O-α-L-rhamnopyranoside (1) and quercetin-3-O-(6-O-benzoyl)-ß-D-glucopyranosyl-(1 → 3)-ß-D-galactopyranoside-7-O-α-L-rhamnopyranoside (2), were isolated together with six known compounds from the seeds of L. sativum. Their structures were elucidated on the basis of spectroscopic analysis and chemical methods. In vitro 1 and 2 inhibited nitric oxide production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, with IC50 values of 25.36 and 25.08 µM, respectively.

[Randomized, double-blind, multi-center, positive parallel control clinical trial of compound Wuzhigan capsules on anemopyretic cold].

Compound Wuzhigan capsules is a compound preparation composed of Wuzhigan, Shidagonglao, Gangmei, Shanzhima. A Randomized, double-blind, multi-center, positive parallel control designed to evaluate the clinical efficacy and safety of compound Wuzhigan capsules on anemopyretic cold. One hundred and twenty anemopyretic cold patients were given compound Wuzhigan capsules (test group), 2 capsules one time, three times a day, 119 patients were given compound Wuzhigan tablets (control group) ,4 tablets one time, three times a day; three days of treatment The study showed, the markedly effective rate and total effective rate respectively were 63. 3% and 80% of the test group. For the control group, the markedly effective rate and total effective rate respectively were 72. 5% and 80. 7%. The difference was not statistically significant. Compound Wuzhigan capsules can reduce the dosage, and get better patient compliance.

[Application of solid-phase extraction column for determination of matrine and oxymatrine in Sophora flavescens].

A Cleanert Alumina-N-SPE column (0.5 g/6 mL) chromatograpy with 5 mL of chloroform-methanol (7: 3) as eluent, instead of aluminum oxide column (100-200 mesh, 5 g, 1 cm) chromatograpy eluted successively with chloroform and the chloroform-methanol (7:3) (20 mL each), was applied to enrich matrine and oxymatrine in Sophora flavescens. Also, the optimization of the HPLC determination conditions with acetonitrile-ethanol absolute-3% phosphoric acid solution (84: 6: 10) as mobile phase, instead of acetonitrile-ethanol absolute -3% Phosphoric acid solution (80: 10: 10) recorded in Chinese Pharmacopoeia 2010 Edition, was more suitable for determination of matrine and oxymatrine in S. flavescens. This method has advantage of reducing sample handling time and solvent volume and increasing the accuracy and feasibility, which can simplify the procedure for determination of matrine and oxymatrine in S. flavescens.

[Study on content of two sesquiterpenes in Alpinia oxyphylla].

OBJECTIVE: To develop an HPLC method for determining oxyphyllenodiol A (1) and teuhetenone A (2) contained in Alpinia oxyphylla and to compare the contents of the two components contained in medicinal materials and prepared herbal medicines in pieces sold in the market and different fractions. METHOD: HPLC and Waters sunfire C18 column (4. 6 mm x 250 mm, 5 microm) were adopted for gradient elute with the mobile phase of acetonitrile and water. The flow rate was 1.0 mL x min(-1) and the detection wavelength was 250 nm. RESULT: 1 and 2 showed a good linear relationship within the range of 0.1296 - 0.8640 microg and 0.1635 - 1.0900 microg respectively, with the average recoveries of 99.08% and 97.80%. Their content ranges were 0.0059% - 0.0149% and 0.0080% - 0.0164% in different samples. The mean value of 1 and 2 were 0.0085% and 0.0104% in the whole fruits, and 0.0137% and 0.0157% in the seeds. They were undetected in the nutshells. CONCLUSION: The method is so precise, accurate and highly reproducible that it can be used to determine the contents of oxyphyllenodiol A and teuhetenone A in A. oxyphylla. The contents of the two components are mainly extracted from the seeds, with certain difference among different samples. There are a higher contents and no significant difference in the salted and raw seeds.

Antihypertensive effects of Hsian-tsao and its active compound in spontaneously hypertensive rats.

The present study aimed to investigate the effects of Hsian-tsao (Mesona procumbens Hemsl.) and its active compound on blood pressure, lipid peroxidation and total antioxidant status of spontaneously hypertensive rats (SHRs). Male SHRs were orally administrated either a water extract of Hsian-tsao (WEHT) (1.0 g/kg) or caffeic acid (CA) [0.1 g/kg of body weight (BW)] on a daily basis for 6 weeks. The results indicated that both hepatic and plasmatic malondialdehyde concentration were increased and total liver glutathione (GSH) levels and antioxidant enzyme activities were decreased in SHRs when compared to the control Wistar Kyoto rats at the end of the trail. In SHRs, oral administration of WEHT or CA for 6 weeks reduced blood pressure as well as plasma and hepatic malondialdehyde levels and increased hepatic antioxidant enzyme activities when compared to SHRs control rats. Reverse transcriptase-polymerase chain reaction results indicated that the changes in hepatic antioxidant enzyme mRNA levels by WEHT or CA were similar to those noted in the enzyme activity levels. The hepatic levels obtained from WEHT or CA-administrated rats had significantly greater oxygen radical absorbance capacity values and total GSH levels than those of control rats. Following oral administration of CA, phenolic acid was detected in the plasma, and C(max) value after 1.0 h administration was 0.92 micromol/L. These findings indicate that a supplement of Hsian-tsao may prevent development of increased blood pressure and enhance the total antioxidant status in vivo.

[Studies on furmaric acid and isofraxidin content in Sarcandra glabra of different provenances].

OBJECTIVE: To study the content variation of furmarid acid and isofraxidin in Sarcandra glabra from 21 different provenances and provide the basis for resource utilization and quality optimization of S. glabra. METHOD: HPLC method was developed to determine the contents of furmarid acid and isofraxidin in 330 samples of S. glabra which were collected respectively from 21 different provenances. RESULT: There were significant differences in the contents of isofraxidin and furmarid acid in S. glabra from different provenances. The contents of isofraxidin and furmarid acid dropped off from low altitude to high altitude, which were also close with longitude and latitude. The content of isofraxidin in S. glabra at central area of natural distribution was the highest. The different parts of the plant had different results, the influence on the contents of the acitive components in stem were more obvious than the leaf. CONCLUSION: This simple, accurate and reproducible method could be use to determine the contents of furmarid acid and isofraxidin in S. glabra. The results represented the status of medicines quality and difference of Chinese S. glabra. These agreed with the traditional views that the medicines quality of Sarcandra glabra in Jiangxi, Fujian, Zhejiang was better. These provenances were considered as important areas of medicines breeding and bases building on S. glabra in future.

[Studies on fingerprinting and identification of main species of Herba Epimedii].

OBJECTIVE: To establish an effective method for the identification of Herba Epimedii. METHOD: The HPLC fingerprints of 8 officinal species including 5 species recorded in 'Chinese Pharmacopoeia', which are Epimedium brevicornu, E. sagittatum, E. pubescens, E. koreanum and E. wushanense, and the other 3 species, E. acuminatum, E. myrianthum and E. leptorrhizum, recorded in 'Guizhou Quality Criteria for Traditional Chinese Medical Material and Nationality Medical Material', and of other 24 species in genus Epimedium in China as well had been analyzed and some fingerprinting features for the identification had been pointed out. RESULT AND CONCLUSION: The fingerprinting features of E. koreanum are rather different from others and can be used for the species identification. There are some common fingerprinting features in the most samples of E. brevicornu, E. acuminatum and E. myrianthum, respectively, and these features are seldom found in the samples of other species. The HPLC fingerprints of E. pubescens and E. leptorrhizum appear steadily among all samples of the species respectively, but it is difficult to distinguish the spcecies from other species with the HPLC fingerprints only. The HPLC fingerprints of E. sagittatum and E. wushanense vary greatly within the species. It is also showed that the quality of Herba Epimedii varied significantly according to the HPLC features.

[Study on total alkaloids content and its influential factors in medicinal materials of Tripterygium].

OBJECTIVE: To investigate the total alkaloids content profile and diversity of medicinal materials of Tripterygium. METHOD: The total alkaloids were extracted by organic solvent and purified by column chromatography (aluminium oxide), and determined by UV spectrometry. RESULT AND CONCLUSION: The total alkaloids content in phloem was 2 to 5 times richer than that in xylem, and their changes had shown concordance. For the different species/types, the content from high to low in order was T. hypoglaucum, Tripterygium F, T. wilfordii and T. regellii, but in each type, the disparity among different populations content was very small. The populations with higher content were in Suichuan, Jiangxi Province, Huangshan, Anhui Province, Jingning, Zhejiang Province and Kunming, Yuannan Province; which came from T. hypoglaucum and Tripterygium F. The degree of difference of total alkaloids content among individuals in each population was different. The populations, which had a rich content diversity, were in Jingning, Zhejiang Province, Taining, Fujian Province and Kunming, Yunnan Province. They will be used for good germplasm selecting. The thickness of medicinal materials did not affect the total alkaloids content in the same plant. The total alkaloids content in xylem changed with seasons: the highest in March, then declining from June to December, the lowest in December, While this did not happen in phloem.

Fast quality control of Herba Epimedii by using Fourier transform infrared spectroscopy.

Herba Epimedii is a well-known traditional Chinese medicine (TCM) having the effect of nourishing the kidney and strengthening the 'Yang'. Its primary effective constituents are considered to be the 8-prenyl flavonols, which can be assorted into 4'-methoxyl-prenylflavonols (MPFs) and 4'-hydroxyl-prenylflavonols (HPFs), according to the group (methoxyl or hydroxyl) located at 4' in their structures. The Fourier transform infrared spectroscopy (FT-IR) has been widely used in the researches of TCMs. In the present study, the FT-IR was attempted to be applied in the quality control of Herba Epimedii. We compared the IR spectra of 17 pure flavonoids, of which eight were derived from Herba Epimedii, and found a characteristic absorption peak at 1259+/-1 cm(-1), corresponding to the MPFs, the major 8-prenyl flavonols in the aerial parts of the Epimedium species. This peak could also be found in the IR spectra of both the herbal samples and their 70% ethanol extracts. Moreover, the intensity of this peak was in the direct correlation with the total content of MPFs. The correlation values, representing the semblance of two spectra, of the IR spectrum of herbal sample and icariin, in the range of 1280-1200 cm(-1), had been established to be a good index for the quality control of the herbs. Accordingly, a correlation value of not less than 0.50 could be used as the essential screening criteria for the herbs. The FT-IR could be used for the fast and effective quality control of Herba Epimedii.