Cell culture medium supplemented with taurine decreases basic charge variant levels of a monoclonal antibody.

OBJECTIVE: To explore the impact of taurine on monoclonal antibody (mAb) basic charge variants in Chinese hamster ovary (CHO) cell culture. RESULTS: In fed-batch culture, adding taurine in the feed medium slightly increased the maximum viable cell density and mAb titers in CHO cells. What's more, taurine significantly decreased the lysine variant and oxidized variant levels, which further decreased basic variant contents from 32 to 27%. The lysine variant content in the taurine culture was approximately 4% lower than that in control condition, which was the main reason for the decrease in basic variants. Real-time PCR and cell-free assay revealed that taurine played a critical role in the upregulation of relative basic carboxypeptidase and stimulating extracellular basic carboxypeptidase activities. CONCLUSION: Taurine exhibits noticeable impact on lower basic charge variants, which are mainly due to the decrease of lysine variant and oxidized protein variants.

[Polydatin induces human cervical cancer cell apoptosis via PI3K/AKT/mTOR signaling pathway].

To observe the effect of polydatin on proliferation and apoptosis of cervical cancer HeLa cells and explore its possible mechanism. The growth inhibitory effect was detected with MTT assay. After HeLa cells were treated with different concentrations (50, 100, 150 µmol•L⁻¹) of polydatin, MTT assay was used to detect the inhibitory effect of polydatin on proliferation of HeLa cells; Acridine orange/ethidium bromide staining was used for morphological changes in apoptotic HeLa cells； Annexin/propidium iodide staining was applied to detect HeLa cell apoptotic rate. In addition, flow cytometry was employed to analyze apoptosis and cell cycle distribution； RT-PCR and Western blot assay were used to detect PI3K, AKT, mTOR, and P70S6K mRNA and protein expression levels. The results showed that polydatin significantly inhibited HeLa cells proliferation in a dose-dependent manner. Polydatin can cause S phase arrest for HeLa cells, promote cell apoptosis and decrease the mRNA and protein expression levels of PI3K, AKT, mTOR and P70S6K. It indicated that polydatin could inhibit proliferation and induce apoptosis of cervical cancer HeLa cells, and the mechanism may be associated with inhibiting the PI3K/AKT/mTOR signaling pathway and suppressing downstream gene expression.

Electroacupuncture activates enteric glial cells and protects the gut barrier in hemorrhaged rats.

AIM: To investigate whether electroacupuncture ST36 activates enteric glial cells, and alleviates gut inflammation and barrier dysfunction following hemorrhagic shock. METHODS: Sprague-Dawley rats were subjected to approximately 45% total blood loss and randomly divided into seven groups: (1) sham: cannulation, but no hemorrhage; (2) subjected to hemorrhagic shock (HS); (3) electroacupuncture (EA) ST36 after hemorrhage; (4) vagotomy (VGX)/EA: VGX before hemorrhage, then EA ST36; (5) VGX: VGX before hemorrhage; (6) α-bungarotoxin (BGT)/EA: intraperitoneal injection of α-BGT before hemorrhage, then EA ST36; and (7) α-BGT group: α-BGT injection before hemorrhage. Morphological changes in enteric glial cells (EGCs) were observed by immunofluorescence, and glial fibrillary acidic protein (GFAP; a protein marker of enteric glial activation) was evaluated using reverse transcriptase polymerase chain reaction and western blot analysis. Intestinal cytokine levels, gut permeability to 4-kDa fluorescein isothiocyanate (FITC)-dextran, and the expression and distribution of tight junction protein zona occludens (ZO)-1 were also determined. RESULTS: EGCs were distorted following hemorrhage and showed morphological abnormalities. EA ST36 attenuated the morphological changes in EGCs at 6 h, as compared with the VGX, α-BGT and HS groups. EA ST36 increased GFAP expression to a greater degree than in the other groups. EA ST36 decreased intestinal permeability to FITC-dextran (760.5 ± 96.43 ng/mL vs 2466.7 ± 131.60 ng/mL, P < 0.05) and preserved ZO-1 protein expression and localization at 6 h after hemorrhage compared with the HS group. However, abdominal VGX and α-BGT treatment weakened or eliminated the effects of EA ST36. EA ST36 reduced tumor necrosis factor-α levels in intestinal homogenates after blood loss, while vagotomy or intraperitoneal injection of α-BGT before EA ST36 abolished its anti-inflammatory effects. CONCLUSION: EA ST36 attenuates hemorrhage-induced intestinal inflammatory insult, and protects the intestinal barrier integrity, partly via activation of EGCs.

Combination treatment with whole body vibration and a kidney-tonifying herbal Fufang prevent osteoporosis in ovariectomized rats.

OBJECTIVE: To assess the ability of whole body vibration (WBV) with the kidney-tonifying herbal Fufang (Bushen Zhuanggu Granules, BZG) to prevent osteoporosis in ovariectomized rats. METHODS: Fifty 6-month-old female Sprague Dawley rats were divided into five groups: sham-operated (SHAM), ovariectomized (OVX), OVX with WBV (OVX + WBV), OVX with BZG (OVX + BZG), OVX with both WBV and BZG (OVX + WBV + BZG). The SHAM group received normal saline. After 12 weeks of treatment, the rats were killed, their serum concentrations of osteopontin (OPN), receptor activator of nuclear factor kappa-B ligand RANKL and bone turnover markers assayed and bone mineral density (BMD), histomorphometry and bone strength evaluated. RESULTS: Concentrations of OPN were significantly lower in the SHAM, OVX + WBV and OVX + WBV + BZG groups at 12 weeks, whereas concentrations of RANKL had decreased significantly in the SHAM, OVX + WBV, OVX + BZG and OVX + WBV + BZG groups. In the OVX + WBV, OVX + BZG and OVX + WBV + BZG groups the amount of bone turnover had been significantly antagonized. Compared with OVX group, BMD, % trabecular area (Tb.Ar), number of trabeculae (Tb.N) and assessed biomechanical variables were higher in OVX+WBV group, whereas and BMD, %Tb.Ar, Tb.N, maximal load and yield load were higher in the OVX + BZG group. All tested indices were significantly lower in the OVX + WBV and OVX + BZG groups than in the OVX + WBV + BZG group. CONCLUSION: Either WBV or BZG alone prevents OVX-induced bone loss. However, BZG enhances the effect of WBV by further enhancing BMD, bone architecture and strength.

Electroacupuncture at Zusanli (ST36) promotes gastric emptying and mucosal blood flow during oral resuscitation of scalded rats with a pyruvate-enriched ORS.

AIM: The aim of this study was to investigate the effect of electroacupuncture at ST36 (EA ST36) on gastric emptying and mucosal blood flow during intragastric resuscitation with pyruvate-enriched oral rehydration solution (Pyr-ORS) in scalded rats. METHODS: The rats were subjected to a 35% total body surface area (TBSA) of scald injury and randomly divided into five groups (N=24) and two subgroups (n=12) in each group. The Pyr-ORS was delivered intragastrically according to the Parkland formula immediately after scalding at a dose of 1 mL kg(-1) %TBSA(-1) in 1 h. In these animals, the bilateral Zusanli points (ST36) were electroacupunctured at a constant voltage (2 mA and 2-100 HZ) for 0.5 h immediately after intragastric resuscitation. At 2 and 4 h after scalding, the gastric emptying rate (GER) and gastric mucosal blood flow (GMBF) were determined, and the motilin levels of the plasma and gastric tissues were also analyzed at two time points, respectively. RESULTS: GER and GMBF were markedly decreased in groups with scalding and resuscitation, compared with the sham groups at two time points (P<0.05), but they were greatly improved in groups byEAST36 at 2 and 4 h after sustaining scald injuries (P<0.05). Bilateral vagotomy further aggravated the reduction of GER and GMBF in scalded rats. EA after gastric vagotomy failed to raise GER and GMBF. Neither EA nor vagotomy had effects on the reduced motilin levels of plasma and gastric tissues in animals after scalding. CONCLUSION: EA ST36 has a significant effect on improving gastric emptying and mucosal ischemia in the oral resuscitation of burn injury, possibly through the activation of a cholinergic nerve-dependent mechanism. In addition, EA ST36 showed no effects on motilin levels, but requires further investigations.

Factors affecting quantity of pollen dispersal of spray cut chrysanthemum (Chrysanthemum morifolium).

BACKGROUND: Spray cut chrysanthemum is a vital flower with high ornamental value and popularity in the world. However, the excessive quantity of pollen dispersal of most spray cut chrysanthemum is an adverse factor during its flowering stage, and can significantly reduce its ornamental value and quickly shorten its vase life. More seriously, excessive pollen grains in the air are usually harmful to people, especially for those with pollen allergies. Therefore, in order to obtain some valuable information for developing spray cut chrysanthemum with less-dispersed or non-dispersed pollen in the future breeding programs, we here investigated the factors affecting quantity of pollen dispersal of spray cut chrysanthemum with four cultivars, i.e. 'Qx-097', 'Noa', 'Qx-115', and 'Kingfisher', that have different quantity of pollen dispersal. RESULTS: 'Qx-097' with high quantity of pollen dispersal has 819 pollen grains per anther, 196.4 disk florets per inflorescence and over 800,000 pollen grains per inflorescence. The corresponding data for 'Noa' with low quantity of pollen dispersal are 406, 175.4 and over 350,000, respectively; and 219, 144.2 and nearly 160,000 for 'Qx-115' without pollen dispersal, respectively. 'Kingfisher' without pollen dispersal has 202.8 disk florets per inflorescence, but its anther has no pollen grains. In addition, 'Qx-097' has a very high degree of anther cracking that nearly causes a complete dispersal of pollen grains from its anthers. 'Noa' has a moderate degree of anther cracking, and pollen grains in its anthers are not completely dispersed. However, the anthers of 'Qx-115' and 'Kingfisher' do not crack at all. Furthermore, microsporogenesis and pollen development are normal in 'Qx-097', whereas many microspores or pollen degenerate in 'Noa', most of them abort in 'Qx-115', and all of them degrade in 'Kingfisher'. CONCLUSIONS: These results suggest that quantity of pollen dispersal in spray cut chrysanthemum are mainly determined by pollen quantity per anther, and capacity of pollen dispersal. Abnormality during microsporogenesis and pollen development significantly affects pollen quantity per anther. Capacity of pollen dispersal is closely related to the degree of anther dehiscence. The entire degeneration of microspore or pollen, or the complete failure of anther dehiscence can cause the complete failure of pollen dispersal.

[Selection of winter plant species for wetlands constructed as sewage treatment systems and evaluation of their wastewater purification potentials].

In order to establish an evaluation system for selection of winter wetland plants possessing high wastewater purification potentials in subtropics areas, designed sewage treatment experiments were carried out by introducing into the constructed wetlands 25 species of winter wetland plants. Cluster analysis was performed by including harmful environment-resistant enzyme and substrate enzyme activities into the commonly applied plant screening and assessment indexes system. The obtained results indicated that there were significant differences among the tested winter plants in their root length and vigor, leaf malonaldehyde (MDA), biomass, average nitrogen and phosphorus concentration and uptake, and urease and phosphoric acid enzyme activities in the root areas. Based on the established evaluation system, the tested plants were clustered into 3 groups. The plants in the 1st group possessing high purification potentials are Oenanthe javanica, Brassicacapestris, Juncus effusu, Saxifragaceae, Iris pseudoacorus, Osmanthus fragrans and Iris ensata; those in the 2nd group possessing moderate purification potentials are Brassica oleracea var acephala, Calendula officinalis, Aucuba japonica, Ligustrum lucidu, Beta vulgaris, Rhododendron simsii and Ilex latifolia; and those in the 3rd group with low purification potentials are Brassica oleracea var acephala, Calistephus chinensis, Rosa chinensis, Antirrhinums, Liriope palatyphylla, Zephyranthes candida, Fatshedera lizei, Petunia hybrida, Ilex quihoui, Dianthus caryophyllus and Loropetalum chinensis.

[Human osteoblast-like cells OS-732 intervened with alcohol and treated with liuweidihuang pill medicated serum].

OBJECTIVE: To investigate the mechanism of osteonecrosis of the femoral head caused by alcohol and the therapeutic effects of the method of invigorating the kidney. METHODS: The osteoblast-like cells were intervened with 0.3, 0.9, 1.5 mol/L alcohol and observed by flow cytometry and light microscopy and electro microscopy. The cells intervened by 0.15 mol/L alcohol were treated with liuweidihuang pill medicated serum and observed by flow cytometry and light microscopy and electro microscopy. RESULTS: Apoptosis rate and the degree of cell necrosis had positive correlation with the concentration and the acting time of alcohol. The polymorphism of cells wer turned into roundness, and the cellular ecphyma was shortened and thinning, ribosome and endocytoplasmic reticulum were diminished. Part of the ruffle in mitochondria was disappeared. After treatment, the cellular ecphyma was recovered and the endoplasmic reticulum and golgi apparatus were visible. CONCLUSION: The osteoblast-like cells could be necrosis and apoptosis by the alcohol and were positive correlation with the intervened concentration and acting time. Chinese medicine with method of invigorating the kidney has effect on rehabilitating the cellular activity.

Treatment of infantile hemangioma with optimal pulse technology.

BACKGROUND AND OBJECTIVE: The treatment of infantile hemangioma must address both the effectiveness of the treatment and have as few adverse events as possible for the patient as a result of the therapy. The intense pulsed light (IPL) source can be useful in this regard in treating infantile hemangioma. IPL with optimal pulse technology (OPT) represents a new generation of IPLs and in this clinical investigation, the efficacy and adverse event profiles of treating infantile hemangiomas with an IPL with OPT will be reviewed. METHODS: A total of 62 patients with infantile hemangiomas were included in this clinical trial. The mean age of the patients was 6 months old. The Fitzpatrick skin types for those enrolled was either Type III or Type IV. Each patient was subjected to a treatment protocol which included four to five IPL treatments at 4-week intervals. The patients were then assessed at 3 months following their last IPL treatment and clinical improvement was determined by comparisons of pre- and post-therapy photographs. The parents of the patients were asked to score their overall satisfaction with the treatments. RESULTS: From the clinical trial presented, 76% of the infantile hemangiomas were noted to improve with great satisfaction in this clinical trial. A clearance rate of more than 80% was observed. Adverse events, as a result of the IPL treatment, was minimal with less than 5% of the treatments resulting in an adverse event, all of which were noted to be transient in nature. No scarring or pigmentary disturbances were seen in any of the patients evaluated. CONCLUSIONS: This new generation IPL with OPT can be considered a safe and effective modality for the treatment of infantile hemangioma. Marked improvement was noted in the majority of study patients and adverse events were noted to be minimal.

Glycomyces endophyticus sp. nov., an endophytic actinomycete isolated from the root of Carex baccans Nees.

An actinomycete, designated strain YIM 56134(T), was isolated from the root of a Chinese medicinal plant, Carex baccans Nees, collected from Yunnan, south-west China, and subjected to a polyphasic taxonomic study. An analysis of 16S rRNA gene sequence similarities showed that strain YIM 56134(T) was a member of the genus Glycomyces, being most closely related to Glycomyces algeriensis NRRL B-16327(T) (99.0 % similarity), Glycomyces lechevalierae DSM 44724(T) (99.0 %), Glycomyces rutgersensis IFO 14488(T) (98.9 %) and Glycomyces harbinensis IFO 14487(T) (98.7 %). Strain YIM 56134(T) could be distinguished from other established Glycomyces species on the basis of relatively low sequence similarity (<97 %). Phenotypic and chemotaxonomic data supported the affiliation of this strain to the genus Glycomyces. The results of DNA-DNA hybridization and some physiological and biochemical tests allowed differentiation of the strain from related Glycomyces species. Therefore, strain YIM 56134(T) represents a novel species of the genus Glycomyces, for which the name Glycomyces endophyticus sp. nov. is proposed. The type strain is YIM 56134(T) (=KCTC 19152(T) =DSM 45002(T)).

Pivotal advance: eosinophils mediate early alum adjuvant-elicited B cell priming and IgM production.

Alum, aluminum-hydroxide-containing compounds, long used as adjuvants in human vaccinations, functions by ill-defined, immunostimulatory mechanisms. Antigen-free alum has been shown to act via a previously unidentified, splenic Gr1(+), IL-4-expressing myeloid cell population to stimulate early B cell priming. We demonstrate that the alum-elicited and -activated splenic myeloid cells are IL-4-expressing eosinophils that function to prime B cell responses. Eosinophils are the principal Gr1(+), IL-4(+) cells in the spleens 6 days following i.p. alum administration. Alum-elicited splenic B cell priming, as evidenced by MHC II cross-linking-mediated calcium mobilization developed in wild-type BALB/c mice, was absent in DeltadblGATA BALB/c eosinophil-deficient mice and could be reconstituted by adoptive eosinophil infusions into the eosinophil-deficient mice. Moreover, early antigen-specific IgM antibody responses in alum-antigen-immunized mice were impaired in eosinophil-deficient mice and were restored with adoptive transfers of eosinophils. Thus, eosinophils, leukocytes of the innate immune system that contain preformed cytokines, including IL-4, have novel, immunomodulatory roles in the initial priming of B cells elicited by the adjuvant alum and in the optimal early B cell generation of antigen-specific IgM.

[Design of a lithium-ion battery recharging circuit used in wireless electrical stimulation apparatus for urinary incontinence].

We introduce in this paper a novel design of the lithium-ion battery automatically-charging circuit used in portable wireless electronic stimulation therapeutic apparatus for urinary incontinence, including its working principles, structure and characteristic.

[Bioactivity of endophytic actinomycetes from medicinal plants and secondary metabolites from strain D62].

It is believed that genetic recombination of the endophytes with the hosts that occurred in evolutionary time could result in some endophytes producing certain phytochemical originally characteristic of the host. Based on this widely accepted hypothesis, there have been increasing research efforts focused on screening for novel natural products from endophytes. In this study, antimicrobial and antitumor activities of 165 actinomycetes isolated from medicinal plants collected from Xishuangbanna were tested by agar diffusion method and WST-8 assay respectively. The results showed that over 42% of the isolates exhibited antagonism against pathogenic strains, and 54.5% displayed excellent inhibition against mouse melanoma cell line B16 or/and human alveolar epithelial cell line A549. These results are superior to those of soil actinomycetes, indicating tremendous potential of endophytic of actinomycetes for exploration. Six compounds that had both antimicrobial and antitumor activities were separated and purified from isolate Streptomyces sp. D62 by resin adsorption, silica-gel column and sephadex chromatography, etc. On the basis of spectral analyses, they were identified as antimycin A4a (1), antimycin A7a (2), antimycin A2a (3), antimycin A1a (4), 10-hydroxy-10-methyl-dodec-2-en-1,4-olide (5) and 6-(2-(4-aminophenyl)-2-oxoethyl)-3,5-dimethyl-tetrahydropyran-2-one(6), with the last one defined as a novel compound. Based on all these results, it is convinced that endophytic actinomycetes are a promising resource for bioactive natural product discovery.

[Clinical observation on treatment of 34 advanced gastric carcinoma patients by chemotherapy of DCF regimen combined with Fuzheng Hewei Decoction].

OBJECTIVE: To evaluate the therapeutic and adverse reactions of integrative Chinese and Western medicine (ICWM) for treatment of advanced gastric carcinoma. METHODS: Sixty-six patients with advanced gastric carcinoma were randomized into two groups. Patients in the treatment group (34 patients) received chemotherapy of DCF (docetaxel/cisplatin/5-fluorouracil) regimen combined with Chinese herbal medicine, and patients in the control group (32 patients) were treated with chemotherapy of the same regimen alone. RESULTS: No significant difference between the two groups was found in response rate (55.9% versus 50.0%), clinical benefiting rate (85.3% versus 81.3%) and half year survival rate (82.4% versus 65.6%); but the 1-year survival rate in the treatment group was superior to that in the control group (52.9% versus 25.0% , P < 0.05). As for the adverse reactions, significant difference between the two groups was shown in incidence of leucopenia, nausea and vomiting, mucosa reaction and fatigue, these reactions occurred lesser in the treatment group (P< 0.05). CONCLUSION: Although treatment of ICWM for advanced gastric carcinoma cannot improve response rate, it could alleviate the adverse reactions of the treatment, improve quality of life and prolong the survival rate in patients.

[Effects of carotenoids extracts from Potamogeton crispus on apoptosis of human hepatoma cell line QGY-7703 in vitro].

Human hepatoma cell line QGY-7703 was treated with the medium containing 10, 20, 40 and 60 micromol/L carotenoids extracts from Potamogoton crispus L. (CEPC) for 48 h, 96 h and 144 h, respectively. The inhibition rates to hepatoma cells were determined by MTT assay. The results showed the average inhibition rates of the three treatments varied from 0.14%-23.07%, 39.59%-70.61% and 71.65%-87.01%, respectively. After hepatoma cells were treated with the medium containing 10, 20 and 40 micromol/L of CEPC for 24 h, 48 h and 72 h, respectively, many typical morphology features of apoptotic cells were observed by Laser Scanning Confocal Microscope (LSCM), such as the distinct decrease in number and volume of cells, the shrinkage and deformation of cells, the shape of cell nucleuses appearing like crescent even piece, the decrease in area of yellow DNA in cell nuclei. The percentage of hepatoma cells in every phase of cell cycle was analyzed by Flow Cytometer (FCM) after being treated with 10 micromol/L and 20 micromol/L CEPC for 48 h. Compared with the control group, the cells in G0/G1 phase increased 23.8% (P<0.01) and 35.6% (P<0.01) respectively, in S phase decreased relevantly, while the cells in G2/M phase had no significant change. The Ca2+ concentration (fluorescence intensity) in cytoplasm of hepatoma cells was determined by LSCM after being treated with 20 micromol/L CEPC for 48 h. The Ca2+ concentration increased significantly (P<0.01) and was 1.5 times that of control group. These results demonstrated that the CEPC inhibited the proliferation of hepatoma cells QGY-7703 significantly in a dose and time-dependent manner in vitro. The hepatoma cells treated with CEPC were evidently arrested at G0/G1 phase in the cell cycle. The concentration of Ca2+ in test group cells cytoplasm increased significantly, it might be the important cause that CEPC induced the apoptosis of hepatoma cells QGY-7703. This paper provided scientific basis for further studying and developing the function and value of carotenoids in Potamogoton crispus L.