Enhanced Production of Hypocrellin A in Submerged Cultures of Shiraia bambusicola by Red Light.

Hypocrellin A (HA), a promising photosensitizer for anticancer photodynamic therapy (PDT), is a fungal perylenequinone pigment from the fruiting body of Shiraia bambusicola, a traditional Chinese medicine for treating skin diseases. The mycelial cultures are becoming a biotechnological alternative for HA production. In this study, light of different wavelengths was investigated to develop an effective eliciting strategy for HA production in the cultures. Under red LED light (627 nm) at 200 lux, the maximum HA production (175.53 mg L-1 ) in mycelium cultures was reached after 8 days, about 3.82-fold of the dark control. Red light not only promoted HA biosynthesis in mycelia (intracellular HA), but also stimulated HA secretion into the medium (extracellular HA). We found 14 of 310 transcripts differentially expressed under red light treatment were possible candidate genes for HA biosynthetic pathway. Gene ontology (GO) analysis revealed that red light treatment could change the gene expressions responsible for HA biosynthesis and the transmembrane activity, suggesting both intracellular HA and its secretion could contribute to the enhancement of total HA production in the cultures. The results provided new insights of red light elicitation and effective strategy for HA production in mycelium cultures.

[Plasmakinetic enucleation of the prostate versus holmium laser enucleation of the prostate for benign prostatic hyperplasia].

OBJECTIVE: To evaluate the clinical effects of plasmakinetic enucleation of the prostate (PKERP) and holmium laser enucleation of the prostate (HoLEP) in the treatment of BPH. METHODS: We retrospectively analyzed the clinical data on 78 BPH patients treated by PKERP (n = 38) or HoLEP (n = 40) from January 2016 to October 2017. We recorded the operation time, intraoperative hemoglobin level, catheter-indwelling time, bladder irrigation time, hospital stay, 6-month postoperative IPSS, quality of life (QOL), maximum urinary flow rate (Qmax), postvoid residual urine (PVR), PSA level, International Index of Erectile Function (IIEF) and postoperative complications, and compared the obtained parameters between the two groups and some of them with the baseline. RESULTS: In comparison with the baseline, both the PKERP and HoLEP groups showed statistically significant differences at 6 months after surgery in the QOL score (4.82 ± 0.56 and 4.70 ± 0.67 vs 2.44 ± 0.69 and 2.92 ± 0.49, P < 0.01), IPSS (19.52 ± 4.96 and 19.44 ± 4.08 vs 9.56 ± 2.5 and 9.81 ± 2.5, P < 0.01), Qmax (ï¼»4.54 ± 1.86ï¼½ and ï¼»4.42 ± 2.89ï¼½ ml/s vs ï¼»17.72 ± 3.46ï¼½ and ï¼»17.27 ± 4.42ï¼½ ml/s, P < 0.01), and PVR (ï¼»83.73±55.33ï¼½ and ï¼»109.65 ± 89.58ï¼½ ml vs ï¼»19.93 ± 11.07ï¼½ and ï¼»18.31 ± 15.03ï¼½ ml, P < 0.01). Statistically significant differences were also found between the PKERP and HoLEP groups in the reduced hemoglobin level (ï¼»21.04 ± 16.96ï¼½ vs ï¼»7.88 ± 6.65ï¼½ g/dl, P = 0.01), catheter-indwelling time (ï¼»7.67 ± 2.27ï¼½ vs ï¼»3.93 ± 2.18ï¼½ d, P = 0.01), bladder irrigation time (ï¼»1.67 ± 0.62ï¼½ vs ï¼»1.3 ± 0.54ï¼½ d, P = 0.05), hospital stay (ï¼»4.22 ± 1.55ï¼½ vs ï¼»3.26 ± 0.9ï¼½ d, P = 0.01), and 6-month postoperative QOL score (ï¼»2.44 ± 0.69ï¼½ vs ï¼»2.92 ± 0.49ï¼½, P = 0.05), but not in the other parameters. CONCLUSIONS: Both PKERP and HoLEP are safe and effective for the treatment of BPH, the former more feasible in primary hospitals, while the latter with the advantages of less bleeding, shorter catheterization and hospital stay, and higher 6-month postoperative QOL score.

Biosynthesis of silver nanoparticles using Artemisia annua callus for inhibiting stem-end bacteria in cut carnation flowers.

A biological method for synthesising silver nanoparticles (AgNPs) was developed using the callus extracts from Artemisia annua L. under sunlight at 25,000 lx. The AgNPs were characterised using transmission electron microscopy, atomic force microscope, X-ray diffraction and Fourier transform infrared spectroscopy. The AgNPs were mostly spherical with the size of 2.1 to 45.2 nm (average 10.9 nm). Pulse treatments of AgNPs at 125, 250 and 500 mg/l for 1 h extended vase life of cut carnation (Dianthus caryophyllus cv. Green Land) flowers. Four dominant bacteria strains Arthrobacter arilaitensis, Kocuria sp., Staphylococcus equorum and Microbacterium oxydans were isolated from the stem-ends of cut D. caryophyllus flowers. AgNP pulse inhibited significantly bacterial growth in vase solution and cut stem ends during all of the vase period. The bacteria related blockage in the stem-ends was significantly alleviated by AgNP pulse because of its higher antibacterial efficacy against the dominant bacteria. In addition, ethylene release of cut carnation flowers was inhibited in response to AgNP pulse. This is the first time that the biologically synthesised AgNPs could be applied as a promising preservative agent for cut carnation flowers.

Antioxidant and DNA Damage Protecting Activity of Exopolysaccharides from the Endophytic Bacterium Bacillus cereus SZ1.

An endophytic bacterium was isolated from the Chinese medicinal plant Artemisia annua L. The phylogenetic and physiological characterization indicated that the isolate, strain SZ-1, was Bacillus cereus. The endophyte could produce an exopolysaccharide (EPS) at 46 mg/L. The 1,1-diphenyl-2-picrylhydracyl (DPPH) radical scavenging activity of the EPS reached more than 50% at 3-5 mg/mL. The EPS was also effective in scavenging superoxide radical in a concentration dependent fashion with an EC50 value of 2.6 mg/mL. The corresponding EC50 for scavenging hydroxyl radical was 3.1 mg/mL. Moreover, phenanthroline-copper complex-mediated chemiluminescent emission of DNA damage was both inhibited and delayed by EPS. The EPS at 0.7-1.7 mg/mL also protected supercoiled DNA strands in plasmid pBR322 against scission induced by Fenton-mediated hydroxyl radical. The preincubation of PC12 cells with the EPS prior to H2O2 exposure increased the cell survival and glutathione (GSH) level and catalase (CAT) activities, and decreased the level of malondialdehyde (MDA) and lactate dehydrogenase (LDH) activity in a dose-dependent manner, suggesting a pronounced protective effect against H2O2-induced cytotoxicity. Our study indicated that the EPS could be useful for preventing oxidative DNA damage and cellular oxidation in pharmaceutical and food industries.

Negative-Pressure Cavitation Extraction of Secoisolariciresinol Diglycoside from Flaxseed Cakes.

The negative-pressure cavitation extraction (NPCE) technique was applied firstly to extract secoisolariciresinol diglucoside (SDG) from flaxseed cakes. The significant extraction parameters were screened by fractional factorial design (FFD). The optimal parameters were determined using the central composite design (CCD) with the two variables, NaOH amount and the liquid/solid ratio. The conditions of the extraction were optimized by using response surface methodology (RSM). Under the optimal conditions, the extraction yield and the extraction purity of SDG was 16.25 mg/g and 3.86%, respectively. The efficiency of NPCE was compared with that of conventional extraction methods. Our results demonstrated that NPCE was comparable to the well-known ultrasound-assisted extraction in term of extraction yield and purity. This extraction technique has advantages of less time-consuming, low solvent usage and high throughput capability.

Antioxidant properties and PC12 cell protective effects of a novel curcumin analogue (2E,6E)-2,6-bis(3,5- dimethoxybenzylidene)cyclohexanone (MCH).

The antioxidative properties of a novel curcumin analogue (2E,6E)-2,6-bis(3,5-dimethoxybenzylidene)cyclohexanone (MCH) were assessed by several in vitro models, including superoxide anion, hydroxyl radical and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and PC12 cell protection from H2O2 damage. MCH displayed superior O2•- quenching abilities compared to curcumin and vitamin C. In vitro stability of MCH was also improved compared with curcumin. Exposure of PC12 cells to 150 µM H2O2 caused a decrease of antioxidant enzyme activities, glutathione (GSH) loss, an increase in malondialdehyde (MDA) level, and leakage of lactate dehydrogenase (LDH), cell apoptosis and reduction in cell viability. Pretreatment of the cells with MCH at 0.63-5.00 µM before H2O2 exposure significantly attenuated those changes in a dose-dependent manner. MCH enhanced cellular expression of transcription factor NF-E2-related factor 2 (Nrf2) at the transcriptional level. Moreover, MCH could mitigate intracellular accumulation of reactive oxygen species (ROS), the loss of mitochondrial membrane potential (MMP), and the increase of cleaved caspase-3 activity induced by H2O2. These results show that MCH protects PC12 cells from H2O2 injury by modulating endogenous antioxidant enzymes, scavenging ROS, activating the Nrf2 cytoprotective pathway and prevention of apoptosis.

Glaucocalyxin A and B-induced cell death is related to GSH perturbation in human leukemia HL-60 cells.

Glaucocalyxin (Gla) A-C are major ent-kauranoid diterpenoids isolated from Rabdosia japonica var. glaucocalyx, a plant used in Chinese traditional medicine as an antitumor and anti-inflammatory agent. The present investigation was carried out to observe whether cellular reduced glutathione (GSH) plays important roles in Gla -induced cytotoxicity. Among major ent-kauranoid diterpenoids isolated, Gla A and B dose-dependently decreased the growth of HL-60 cells with an IC50 of approximately 6.15 and 5.86 µM at 24 h, respectively. Both Gla A and B could induce apoptosis, G2/M-phase cycle arrest, DNA damage and the accumulation of reactive oxygen species (ROS) in HL-60 cells. Moreover, Gla A, B caused rapid decrease of the intracellular GSH content, while inhibition of cellular GSH synthesis by buthionine sulfoximine (BSO) augmented the induced cytotoxicity and apoptosis in HL-60 cells. On the other hand, the administration of GSH or GSH precursor N-acetyl-cysteine (NAC) could rescue Gla A, B-depleted cellular GSH, and abrogate the induced cytotoxicity, G2/M-phase cycle arrest, DNA damage and ROS accumulation in HL-60 cells. Furthermore, Gla A, B decreased the activity of the GSH-related enzymes including glutathione reductase (GR) and glutathione peroxidase (GPX). These data suggest that the intracellular GSH redox system plays important roles in regulating the Gla A, B-induced cytotoxicity on HL-60 cells.

Nitric oxide elicitation for secondary metabolite production in cultured plant cells.

Nitric oxide (NO) is an important signal molecule in stress responses. Accumulation of secondary metabolites often occurs in plants subjected to stresses including various elicitors or signal molecules. NO has been reported to play important roles in elicitor-induced secondary metabolite production in tissue and cell cultures of medicinal plants. Better understanding of NO role in the biosynthesis of such metabolites is very important for optimizing the commercial production of those pharmaceutically significant secondary metabolites. This paper summarizes progress made on several aspects of NO signal leading to the production of plant secondary metabolites, including various abiotic and biotic elicitors that induce NO production, elicitor-triggered NO generation cascades, the impact of NO on growth development and programmed cell death in medicinal plants, and NO-mediated regulation of the biosynthetic pathways of such metabolites. Cross-talks among NO signaling and reactive oxygen species, salicylic acid, and jasmonic acid are discussed. Some perspectives on the application of NO donors for induction of the secondary metabolite accumulation in plant cultures are also presented.

[Observation on therapeutic effect of peripheral facial paralysis treated by string puncture combined with electroacupuncture].

OBJECTIVE: To explore the effective treatment of peripheral facial paralysis. METHODS: Ninety-five cases of peripheral facial paralysis were randomly divided into an observation group (48 cases) and a control group (47 cases). In observation group, the acupoints located on paralyzed muscle, such as Yangbai (GB 14), Taiyang (EX-HN 5), Tongziliao (GB 1), Jiache (ST 6) and Dicang (ST 4), etc. were selected; string puncture was applied and the body of needles went through the muscular layer at 40 degrees-60 degrees angle with the direction of muscle fiber; Yangbai (GB 14), Taiyang (EX-HN 5) etc. were selected for electroacupuncture. In control group, the same acupoints and routine manipulations were applied. The clinical therapeutic effects, courses and the House-Brackmann (H-B) facial nerve function grading of both groups were compared. RESULTS: The total effective rate in observation group was 100.0% (47/47) which was superior to 93.6% (44/47) in control group (P < 0.01); the average effective, marked effective and cured durations of cured patients in observation group were much shorter than those in control group (all P < 0.01); the H-B facial nerve function grading of both groups were improved after treatment (all P < 0.01), and more significant in observation group (P < 0.01). CONCLUSION: By string puncture therapy, peripheral facial paralysis is treated more effective than that by routine puncture, and the curative courses are obviously shortened.

Glaucocalyxin A induces apoptosis in human leukemia HL-60 cells through mitochondria-mediated death pathway.

Glaucocalyxin A (GLA) is a biologically active ent-kauranoid diterpenoid isolated from Rabdosia japonica var. glaucocalyx, a traditional Chinese medicinal herb, which has been shown to inhibit tumor cell proliferation. However, the mechanism underlying GLA-induced cytotoxicity remains unclear. In this study, we focused on the effect of GLA induction on apoptosis, the mitochondria-mediated death pathway and the accumulation of reactive oxygen species (ROS) in human leukemia cells (HL-60). GLA could induce a dose-dependent apoptosis in HL-60 cells as characterized by cell morphology, DNA fragmentation, activation of caspase-3, -9 and an increased expression ratio of Bax/Bcl-2. The mitochondrial membrane potential (Δψ(m)) loss and cytochrome c release from mitochondria to cytosol were observed during the induction. Moreover, GLA caused a time- and dose-dependent elevation of intracellular ROS level in HL-60 cells, and N-acetyl-l-cysteine (NAC, a well-known antioxidant) could block GLA-induced ROS generation and apoptosis. These data suggest that GLA induces apoptosis in HL-60 cells through ROS-dependent mitochondrial dysfunction pathway.

Tanshinone biosynthesis in Salvia miltiorrhiza and production in plant tissue cultures.

Salvia miltiorrhiza Bunge (Lamiaceae) root, generally called Danshen, is an important herb in Chinese medicine widely used for treatment of cardiovascular diseases. Diterpenoid tanshinons are major bioactive constituents of Danshen with notable pharmacological activities and the potential as new drug candidates against some important human diseases. The importance of Danshen for traditional and modern medicines has motivated the research interest over two decades in the biosynthesis and biotechnological production of tanshinones. Although diterpenes in plants are presumably derived from the non-mevalonate (MVA) pathway, tanshinone biosynthesis in S. miltiorrhiza may also depend on the MVA pathway based on some key enzymes and genes detected in the early steps of these pathways. Plant tissue cultures are the major biotechnological processes for rapid production of tanshinones and other bioactive compounds in the herb. Various in vitro cultures of S. miltiorrhiza have been established, including cell suspension, adventitious root, and hairy root cultures, which can accumulate the major tanshinones as in the plant roots. Tanshinone production in cell and hairy root cultures has been dramatically enhanced with various strategies, including medium optimization, elicitor stimulation, and nutrient feeding operations. This review will summarize the above developments and also provide our views on future trends.

Effect of Al2O3 mole fraction and cooling method on vitrification of an artificial hazardous material. Part 1: variation of crystalline phases and slag structures.

This study investigated how Al ions affect slag structure. During vitrification, pure Al(2)O(3), CaO, and SiO(2) served as the encapsulation phases with the use of Al mol% as an operating parameter. All specimens with the same basicity (mass ratio of CaO to SiO(2)) of 2/3 were vitrified at 1400 degrees C and cooled by air cooling or water quenching. XRD was used to measure the volume fractions of crystalline and amorphous phases. In a non-Al environment, CaSiO(3) was formed in air-cooled and water-quenched slags. With the addition of Al(2)O(3), no crystalline phases were observed in water-quenched slags. With the increase of Al mol% in specimens, the Al ions in air-cooled slags initially acted as an intermediate linking one tetrahedron chain to another and reducing the amount of crystalline phase, then behaved as a network former making the slags amorphous, and finally replaced Si ions in silicate frames to generate a large amount of CaAl(2)Si(2)O(8). Air cooling improved the formation of crystallize structures with more leachable metal ions. A highly crystallized Al-framed structure is not suitable for encapsulating hazardous metals in vitrified slags.

Effect of Al2O3 mole fraction and cooling method on vitrification of an artificial hazardous material. Part 2: encapsulation of metals and resistance to acid.

This study investigated the encapsulation of metals in vitrified slags using a sequential extraction procedure and how Al ions and slag structure affected the encapsulation of heavy metals. It is found that the substitution of Al ions for Si ions weakened the encapsulation of the glass matrix due to the relatively weaker single bond strength of Al-O. In addition, the substitution also governed the phase distribution of metals that tended to stay slags. In comparison to air cooled slags, water quenched slags were more amorphous and offered relatively better encapsulation of metals. These findings were also supported by the SEM observation and XRD analysis. Although Al(2)O(3) may connect the non-bridging oxygen and polymerize the slag structure, the excess addition of Al(2)O(3) reduced the metal encapsulation and acid resistance of slags.

[Relevant low toxicities with rhG-CSF mobilized and cryopreserved autologous peripheral blood stem cell return infusions in children].

The purpose of this study was to evaluate the safety of cryopreserved and thawed peripheral blood stem cell (PBSC) fractionated return infusions in children. 35 children patients with malignant tumors (13 acute leukaemias, 15 neuroblastomas and 7 malignant lymphomas) received fractionated return infusions of cryopreserved stem cells after undergoing high-dose chemotherapy without or with total body irradiation. The toxicities of 70 return infusions were evaluated. All patients were mobilized by chemotherapy plus recombination human granulocyte colony-stimulating factor (rhG-CSF), and then PBSCs were collected by a separator CS-3000 plus or COBE spectra-4. The grafts were cryopreserved in 10% dimethyl sulfoxide (DMSD) and stored in liquid nitrogen. There were totally 70 PBSC transfusions. The total volume of PBSCs transfused: 190 - 420 ml (265 +/- 73 ml or 13.7 +/- 4.2 ml/kg) with a mean of (4.43 +/- 1.91) x 10(8)/kg of PBSCs, and 0.94 +/- 0.18 g/kg of DMSO. The single dose: 90 - 300 ml (132 +/- 37 ml or 6.6 +/- 5.2 ml/kg) with a mean of 0.68 +/- 0.12 g/kg of DMSO. Symptoms occurring during the infusions were recorded. All patients were monitored for 24 hours after infusion. Pulse, blood pressure, body temperature, and respiratory rate were recorded every 15 minutes. At four hours before and 8 hours after infusion, urinalysis was performed. Serum potassium, sodium, creatinine, total bilirubin, aspartate amino transferase (AST), and alanine amino transferase (ALT) levels were examined within 24 hours before and after the first infusion. The results showed that the toxicities observed included hemoglobinuria in 54 return infusions (77.1%), headache in 28 (40.0%), nausea in 24 (34.3%), vomiting in 17 (24.3%), and abdominal pain in 8 (11.4%). Patients who received a graft > 200 ml tended to have a higher frequency of hemoglobinuria, headache, nausea, vomiting, or abdominal pain (P<0.01), and they disappeared quickly, too. Total bilirubin increased after the first return infusion (P<0.01), and there was a significant correlation between the volume of infusion and the degree of total bilirubin increase (r=0.8977, P<0.01). No renal failure or shock occurred. It is concluded that transient hemoglobinuria, headache, nausea, vomiting, and abdominal pain are common toxicities associated with PBSC autograft, and these toxicities are related with a single volume of PBSCs transfused. Total bilirubin increase is correlated with the volume of infusion. In a word, the toxicity is less frequent and lower severe in children with fractionated infusions of cryopreserved peripheral blood stem cell.

[Long-term effect of high dose chemotherapy combined with stem cell transplantation on stage IV neuroblastoma in children].

OBJECTIVE: Neuroblastoma is a highly malignant tumor. Stage IV neuroblastoma has a very poor long-term outcome by conventional chemotherapy and surgery and better therapies are essential. This study aimed to explore the long-term effect of high dose induction chemotherapy combined with autologous peripheral blood stem cell transplantation and 13-cis retinoid acid treatment on stage IV neuroblastoma in children. METHODS: Twenty-eight children with stage IV neuroblastoma, aged 2.1-11.5 years (mean 3.3 +/- 1.9 years), were employed for the study. Primary sites of the tumors included adrenal (n=23), chest (n=3), chest-abdomen (n=1) and sacrum (n=1). Before autologous peripheral blood stem cell transplantation the patients received 6 courses of intensive induction chemotherapy. During chemotherapy the autologous peripheral blood stem cells were harvested and the tumor excision was done. After transplantation the local radiation and 13-cis retinoid acid therapy were administered. RESULTS: After 6 courses of induction chemotherapy 13 patients got complete remission (CR), 11 got partial remission (PR), and 4 had no response. The 24 patients who received CR or PR completed the full therapy. A 3.5 +/- 0.7 years follow-up showed that the 4-year event-free survival of the CR and PR patients was 29.2%. The median no-relapse survival time in CR patients was 4.1 +/- 0.7 years but 2.8 +/- 0.5 years in PR patients (t= 3.9, P < 0.01). CONCLUSIONS: High dose chemotherapy combined with autologous peripheral stem cell transplantation and 13 cis-retinoid acid treatment can improve the long-term outcome of patients with stage IV neuroblastoma. The patients in CR before transplantation had better outcomes than those in PR.

[Treatment of high-risk neuroblastoma with intensive chemotherapy, autologous peripheral blood stem cell transplantation, and 13-cis-retinoic acid].

OBJECTIVE: The prognosis for neuroblastoma in advanced stage is still poor, even under conventional chemotherapy. This study aimed to investigate if very high dose chemotherapy in conjunction with autologous peripheral blood stem cell transplantation and 13-cis-retinoic acid could get excellent results in children with high risk neuroblastoma. METHODS: Six children, aged from 4 to 8 years, with stage IV neuroblastoma were included in the study. The duration of the illness before admission was 1 to 12 months. Primary sites of the diseases were in the abdominal cavity (n = 5) and thoracic cavity (n = 1). All of patients had bone marrow metastasis, and one had multiple bone metastasis and orbital metastasis. All of the patients received very high dose chemotherapy, surgery, local radiation (20-30 Gy), and autologous peripheral blood stem cell transplantation as well as 13-cis retinoic acid. Induction chemotherapy included vincristine 0.67 mg/(m2 x 24 h, x 3), cyclophosphamide 2.1 g/(m2 x 24 h, x 2) and doxorubicin 25 mg/(m2 x 24 h, x 3) for 4 courses. Drugs were given as 24 hour-continuous intravenous infusion. Etopside 200 mg/(m2 x 24 h, x 3) and cisplatin 50 mg/(m2 x 24 h, x 3) were given for 2 courses. Conditioning regimen included carboplatin 400 mg/(m2.d) for 4 days, etoposide 300 mg/(m2.d) for 4 days and melphalan 70 mg/(m2.d) for 3 days. 13-cis retinoic acid 160 mg/(m2.d) started on +59 days for 6 courses, each course including 14 days therapy and 14 days rest. RESULTS: Six patients got a complete response before stem cell transplantation. Their bone marrow metastasis disappeared and so did bone and orbital metastasis. However, marrow suppression due to very high dose chemotherapy occurred in all of the patients, which lasted for 3-4 weeks for peripheral leukocyte recovery. Fever occurred after they finished 1/3 course of chemotherapy. Infection, however, was cured with the use of Fortum and Imipenem, ect. Autologous peripheral blood stem cell transplantation was initiated and successful in all cases. Follow-up studies revealed that all the patients were in CR status 4-18 months after transplant, and the cardiac and liver and renal functions were normal. Meanwhile, bone marrow was recovered or in the process of recovery. CONCLUSION: The new strategies focused on very high dose chemotherapy, autologous peripheral blood stem cell transplantation and biological therapy might be a good option for patients with advance neuroblastoma.