[Purification and component identification of total proanthocyanidins in Choerospondias axillaris pericarp].

The present study determined the quantitative markers of total proanthocyanidins in the purification of the industrial waste Choerospondias axillaris pericarp based on the comparison results of high-performance liquid chromatography(HPLC) and mass spectrometry(MS) and optimized the purification process with two stable procyanidins as markers. The adsorption and desorption of five different macroporous adsorption resins, the static adsorption kinetics curve of NKA-Ⅱ resin, the maximum sample load, and the gradient elution were investigated. The UPLC-Q-TOF-MS/MS was employed for qualitative analysis of the newly-prepared total proanthocyanidins of C. axillaris pericarp. As revealed by the results, NKA-Ⅱ resin displayed strong adsorption and desorption toward total proanthocyanidins. The sample solution(50 mg·mL~(-1)) was prepared from 70% ethanol crude extract of C. axillaris pericarp dissolved in water and 7-fold BV of the sample solution was loaded, followed by static adsorption for 12 h. After 8-fold BV of distilled water and 6-fold BV of 10% ethanol were employed to remove impurities, the solution was eluted with 8-fold BV of 50% ethanol, concentrated, and dried under reduced pressure, and purified total proanthocyanidin powder was therefore obtained. Measured by vanillin-hydrochloric acid method, the purity and transfer rate of total proanthocyanidins were 47.67% and 59.92%, respectively, indicating the feasibi-lity of the optimized process. UPLC-Q-TOF-MS/MS qualitative analysis identified 16 procyanidins in C. axillaris total proanthocyanidins. The optimized purification process is simple in operation and accurate in component identification, and it can be applied to the process investigation of a class of components that are difficult to be separated and purified. It can also provide technical support and research ideas for the comprehensive utilization of industrial waste.

[Comparative of functional components, antioxidant and α-glucosidase inhibition activities between Choerospondias axillaris fruit peel vinegar and apple vinegar].

Based on the idea of plant metabolomics, ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) was used to compare the chemical composition between 6 batches of fruit vinegar brewed from Choerospondias axillaris fruit peel and 6 batches of apple vinegar purchased from 3 companies. Antioxidant and α-glucosidase inhibition activities were also tested in vitro. A total of 43 compounds were identified by reference substance, liquid chromatography-mass spectrometry(LC-MS/MS) fragmentation information or literature data. A total of 40 compounds were identified in the C. axillaris fruit peel vinegar. A total of 16 compounds were identified in apple vinegar. There were 13 common ingredients including organic acids and esters such as citric acid, 2-isopropyl malic acid, and triethyl citrate. The results of partial leastsquares-discriminant analysis(PLS-DA) indicated that they had 33 significantly different compounds such as proanthocyanidin oligomer, quercetin-3-O-rhamnoside and heptadecanoic acid. The proanthocyanidins and flavonoid glycosides in C. axillaris peel vinegar were more abundant than apple vinegar, so it had better health function than ordinary fruit vinegar. The results showed that C. axillaris fruit peel vinegar had stronger antioxidant and α-glucosidase inhibition activities in vitro. The vinegar brewed from waste C. axillaris fruit peel had more chemical ingredients than the apple vinegar. C. axillaris fruit peel vinegar had better biological activity and health function, so it had good development prospect. This study provided the scientific evidence for exploiting the C. axillaris fruit peel into high value-added products. It also provided ideas for the comprehensive development and utilization of similar Chinese medicine waste.

High-level expression and purification of the major house dust mite allergen Der p 2 in Escherichia coli.

Der p 2, a major allergen derived from the house dust mite Dermatophagoides pteronyssinus, is one of the most clinically relevant allergens worldwide. Recombinant Der p 2 (rDer p 2) is useful in clinical diagnosis and disease-specific immunotherapy. However, previous studies showed that Der p 2 can only be expressed in Escherichia coli (E. coli) cells as inclusion bodies, thus protein refolding is required to obtain functional products. Here we report a new method to produce biologically active Der p 2 protein in E. coli. N-terminal hexahistidine- and trigger factor (TF)-tagged Der p 2 was expressed in soluble form in E. coli and purified using a combination of chromatography processes. This procedure produced milligram-level high purity Der p 2 per liter of bacterial culture. Moreover, far-UV region circular dichroism (CD) analysis and serum specific IgE reactivity test demonstrated that the secondary structure and IgE reactivity properties of rDer p 2 produced in our study were almost identical to those of natural Der p 2 (nDer p 2). In conclusion, the method developed in this work provides a useful tool for the production of immunologically active recombinant Der p 2 for clinical applications.

[Studies on agronomic traits of seedlings from different F1 generations of Dendrobium officinale].

OBJECTIVE: To select superior mating combinations from different F1 generations of Dendrobium officinale. METHOD: Twenty-six superior parents from four provenances were selected to make up 26 mating combinations. The agronomic traits of different F1 generation were measured, counted, and analyzed by DPS software. RESULT: There were significant differences between different mating combinations among F1 generations. The variation among full-sib families was significantly higher than that among half-sib families, while the variation inside full-sib families was significantly lower than that inside half-sib families. Twelve agronomic traits were simplified into four principal components, of which the accumulative contribution rate was 84.127 3%. Twenty-six mating combinations have been divided into six groups, plants from the fifth and sixth group grew much better than the others. CONCLUSION: The selection of parents should be emphasized by using hybrid vigor. There were higher genetic gains and superior uniformity inside families when selection and breeding occurs in full-sib families than in half-sib families. Eight superior full-sib families (83 x 34, 66 x 9, 68 x 2, 91 x 69, 66 x 65, 69 x 91, 17 x 66, 66 x 17) have been selected which showed a good agronomic traits of seedlings.

Effects of Persimmon Tannin-Aloe vera Composite on Cytotoxic Activities, and Radioprotection Against X-rays Irradiated in Human Hepatoma and Hepatic Cells.

A persimmon tannin-Aloe vera composite powder (PT-A) was investigated for its capacity to protect against ionizing radiation. Human hepatic cells (L02 cells) and human hepatoma cells (HepG2 cells) were pretreated with different concentrations of PT-A or the single compounds (PT or Aloe vera) and radiated with X-rays. After radiation and post-incubation for 12 h or 24 h, the cell viability, apoptosis, and reactive oxygen species (ROS) production were analyzed by Cell Counting Kit 8 (CCK-8), 2',7'-dichlorfluorescein diacetate (DCFH-DA) staining, and Hoechst 33258 staining/flow cytometry, respectively. CCK-8 results illustrated that the optimal radiation dose L02 cells was 8 Gy for L02 cells, and the cell activity was 71.72% (IC50 = 412.1 µg/mL) after post-radiation incubation of 12 h. For HepG2 cells, the optimal radiation dose was 8 Gy, and the cell activity was 62.37% (IC50 = 213.0 µg/mL). The cell apoptotic rate was the lowest at a PT-A concentration of 200 µg/mL in L02 cells (4.32%, P < 0.05), and at 100 µg/mL in HepG2 cells (9.80%, P < 0.05). ROS production induced by radiation could be effectively inhibited by 200 µg/mL of PT-A in L02 cells, and by 100 µg/mL of PT-A in HepG2 cells. The PT-A composite has good radioprotective effects on cell vitality and apoptosis of X-rays radiation exposure towards L02 cells and HepG2 cells compared to the persimmon tannin or Aloe vera. Therefore, PT-A composite might be useful as a natural, harmless anti-ionizing radiation agent, and has various clinical application prospects in future.

[The estrogenic effect of formononetin and its effect on the expression of rats' atrium estrogen receptors].

OBJECTIVE: To observe the estrogenic effect of formononetin and its effect on the expressions of atrial estrogen receptor subtypes alpha and beta (ERalpha and ERbeta). METHODS: 50 femal rats were randomly divided into five groups: sham group, model group, nilestriol group, formononetin groups of low and high dose. Rats in sham group were cut a piece of fat before closing the abdomen, the others were ovariectomized. Vaginal exfoliated cell were observed from the fifth day to the tenth after operation to test if the model is successful. The sham and model group were given nomal saline in 10 mL/kg by gavage, the remaining three groups were given nilestriol 2.5 mg/(kg x w), low [20 mg/(kg x d) land high dose [100 mg/(kg x d)) of formononetin by gavage respectively. In the 8th week, vaginal exfoliated cell were observed, then decapitated the rats, removed the uterus, weighed and take wright staining microscopy. The relative expressions of ERalpha and ERbeta of right atrium were detected by RT-PCR. RESULTS: The vaginal cells exhibit a change of estrus after had been fed with high dose of formononetin after 8 weeks. Formononetin increase the uterus coefficient and the expression of atrial ERbeta (P < 0.01), but it dose not have any effect on the expression of ERalpha (P > 0.05). CONCLUSION: Formononetin have estrogenic effect in ovariectomized rats, and it can markedly upregulate the expression of rats' atrial ERbeta.

[Study on stray light in self-reversal background correction for atomic absorption spectrometry--the properties of stray light and its influence on background correction].

In self-reversal background correction the particularity of stray light that is not absorbed by the sample matrix is different from that in deuterium lamp and Zeeman background correction. The stray light is evaluated using the cut-off solution method and its effects were studied by means of simulation background absorption using potassium dichromate solution molecular absorption. Experiments confirmed that the stray light due to light source for the sample beam (Conventional pulse, low Current) and reference beam (narrow pulse, high current) is obviously different, with the former being higher than the latter. Therefore, the difference is an important source in self-reversal background correction errors and the errors can not be ignored in high background correction. The present paper also showed the relationships among stray light and lamp current, light energy, and spectral bandwidth. The results show that the ratio of stray light is much related to the lamp current, and the larger the lamp current, the smaller the ratio of stray light. The ratio of stray light that changes with lamp current is not directly related to the energy of analytical line. Stray light ratio decreases with the decrease in spectral bandwidth and its character is similar to continuous spectrum.

[Effect of astragalus injection on immune function in patients with congestive heart failure].

OBJECTIVE: To study the effect of Astragalus Injection (AI) on the humoral immunity (IgG, IgA and IgM), cellular immunity (T-lymphocyte subsets) and soluble interleukin-2 receptor (sIL-2R) in patients with congestive heart failure (CHF). METHODS: Sixty-two in-patients with CHF, whose heart function belonged to NYHA grade II-IV, were randomly divided into two groups. The treated group was treated with AI 30 ml (equivalent to 60 g crude drug), and the control group was treated by nitroglycerine injection 10 mg, the drugs were administered respectively by adding in 5% glucose solution 500 ml for intravenous dripping, once a day, 20 days as one therapeutic course. Venous blood from cubital vein was collected before and after treatment to detect the IgG, IgA, IgM, T-lymphocyte subsets and sIL-2R, and the clinical effect of treatment was evaluated. RESULTS: The clinical heart function markedly improved rate and total effective rate in the treated group was 25.8% and 74.2% respectively, significantly better than those in the control group respectively (P < 0.05 or P < 0.01), the left ventricular ejecting fraction (LVEF) and end syctolic volume (ESV) were improved in both groups (P < 0.05, P < 0.01), and the improvement in the treated group was superior to that in the control group (P < 0.05). In the treated group after treatment, the CD4 level and CD4/CD8 ratio increased (P < 0.05), levels of sIL-2R, IgG and IgA lowered (P < 0.05) significantly, while those in the control group were not changed significantly (P > 0.05). CONCLUSION: AI could improve the immune function of CHF patients, and can be taken as an important auxiliary treatment for CHF.