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Background: The non-cultured epidermal cell suspension method is a well-established but tedious grafting modality in the management of stable vitiligo. Recently a more user-friendly automated epidermal harvesting system has been introduced. Aim: This was a pilot study to compare the efficacy and safety outcomes of the above two grafting procedures. Study design: The study was a single-blinded split-body randomised controlled trial. After scientific and ethical clearance, the trial was registered with CTRI (CTRI/2018/05/014225). Thirty consenting patients of stable vitiligo with 60 near-symmetrical patches were recruited. Block randomisation was done using computer-generated randomisation software and each patch was allocated either of the two grafting modalities. Efficacy was assessed by the Physician Global Assessment Scale on serial images and pain by the Numerical Rating Pain Scale. Results and conclusion: The non-cultured epidermal cell suspension was found to be an overall statistically superior technique to the automated epidermal harvesting system in terms of efficacy (re-pigmentation). Both donor and recipient site complications were significantly less with the automated epidermal harvesting system grafting and this method had the distinct advantage of being a painless and easy technique with minimal recovery time. A novel observation was that a good colour match and near-complete re-pigmentation occurred in patients with a darker skin colour with both techniques. Limitations: The main limitation of our study was the small sample size. Also, the size of the treated patches was limited such that they could be covered by the 5 × 5 cm size of the automated epidermal harvesting system blade. However, a larger area can be covered with multiple sessions.
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The aim of this study was to produce Erns protein of bovine viral diarrhea virus (BVDV) by using suspensively cultured CHO cells expression system and to analyze the immunogenicity of the purified Erns protein. In this study, the recombinant eukaryotic expression plasmid pcDNA3.1-BVDV-Erns was constructed based on the gene sequence of BVDV-1 NADL strain. The Erns protein was secreted and expressed in cells supernatant after transfecting the recombinant expression plasmid pcDNA3.1-BVDV-Erns into CHO cells. The expression and purification of the Erns protein was analyzed by SDS-PAGE, the reactivity was determined with anti-His monoclonal antibodies and BVDV positive serum with Western blotting. Immunogenicity analysis of the Erns protein was determined after immunizing New Zealand white rabbits, and the serum antibodies were tested by indirect ELISA (iELISA) and indirect immunofluorescence (IFA). The serum neutralizing titer of the immunized rabbits was determined by virus neutralization test. The concentration of the purified Erns protein was up to 0.886 mg/mL by BCA protein quantification kit. The results showed that the Erns protein could be detected with anti-His monoclonal antibodies and anti-BVDV sera. Serum antibodies could be detected by iELISA on the 7th day post-prime immunization, and the antibody level was maintained at a high titer until the 28th day post-immunization. The antibody titer was 1:128 000. Furthermore, the expression of the Erns protein in BVDV-infected MDBK cells could be detected with immunized rabbits sera by IFA. Moreover, antigen-specific neutralizing antibodies of 2.71 log10 was induced in rabbits. In this study, purified BVDV Erns protein was successfully produced using CHO suspension culture system, and the recombinant protein was proved to have a good immunogenicity, which may facilitate the development of BVD diagnosis method and novel subunit vaccine.
Subject(s)
Rabbits , Animals , Cricetinae , Cricetulus , CHO Cells , Antibodies, Viral , Diarrhea Viruses, Bovine Viral/genetics , Antibodies, Monoclonal/genetics , Diarrhea , Viral Vaccines/geneticsABSTRACT
【Objective】 To investigate the incidence of clinical massive blood transfusion in hospitals, the proportion of departments conducted massive blood transfusion and the current situation of component transfusion, so as to provide a theoretical basis for medical decision-making and further research on massive blood transfusion. 【Methods】 The basic clinical data and transfusion of blood components were retrospectively collected from 489 patients (514 occasions) who received massive blood transfusion at Sun Yat-sen Memorial Hospital of Sun Yat-sen University from Jan. 1 2014 to Dec. 31 2018. 【Results】 The incidence of massive blood transfusion during the 5-year period was 1.2/1 000 inpatients (95%CI: 1.1-1.3), and the 30-day all-cause mortality was 21.88%; in the departments where massive blood transfusion occurred, the mortality rate was the highest in the trauma emergency department (60%), followed by intensive care unit (56.25%) and other surgery department (46.67%), while there was no death in the obstetric department. All patients received red blood cells [median 14 U (11.5-19.13)] and plasma [median 1 600 mL (1 200-2 200)], of which 47% received platelet [median 0 U (0-10)] and 32.68% received cryoprecipitate [0 U (0-10)]. The results of logistics regression analysis of all-cause mortality risk showed that compared with the youth group, the risk of all-cause death at 30 days of elderly patients over 65 years old (65 80 years old: OR=7.563, 95%CI=[1.587, 36.049], P<0.05) and 24-hour RBC infusion volume greater than 18 U (18≤RBC<27: OR=2.948 95%CI=[1.592, 5.462], P<0.05; RBC≥28: OR=3.992, 95%CI=[1.178, 13.536], P<0.05) was higher. 【Conclusion】 A dynamic definition should be included in massive transfusion studies. If only a 24-hour RBC infusion volume ≥18 U was used as the mass transfusion definition, about 68% of cases would be lost. The mortality rate of patients with massive blood transfusion was higher, and the incidence of massive blood transfusion was higher in the departments of cardiac surgery, general surgery and orthopedics surgery. More attention should be paid to the increasing number of female patients with massive blood transfusion. In addition, the risk of 30-day all-cause death was highest in elderly patients over 65 years of age and those with a 24-hour erythrocyte transfusion level of ≥18 U.
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BACKGROUND: Flow cytometry is currently the advanced cell analysis technique with single cell suspension as a research basis, but there is no report on the preparation method of single cell suspension of diaphragm tissue. OBJECTIVE: To explore the feasibility of preparing single cell suspension of rat diaphragm tissue by mechanical grinding method and mechanical-enzymatic digestion method and to compare cell number and viability of the cells obtained using different methods. METHODS: The fresh diaphragm tissues of Sprague-Dawley rats were harvested. Based on the mechanical method, trypsin, collagenase I, collagenase II, collagenase IV and their different combinations were used to digest and prepare the single cell suspension of diaphragm tissues. Cell morphology was observed; cell number and viability were determined by trypan blue staining. The living cells, inactivated cells, and cell aggregates were counted, and cell survival rate and concentration of the single cell suspension were calculated. Statistical analyses were performed thereafter. RESULTS AND CONCLUSION: (1) The single cell suspension with better cell dispersion, more complete morphology, clearer boundary, fewer impurities and cell debris, and cleaner background were obtained by mechanical-enzymatic digestion compared with mechanical grinding method. (2) The single cell suspension prepared by simple mechanical grinding method has low number of living cells, high number of inactivated cells, low survival rate and many cell aggregates. (3) The number of living cells and concentration of single cell suspension obtained by same-volume addition of trypsin, collagenase I and collagenase IV mixed enzymes based on the mechanical grinding method were the highest, with (1.0-2.0)×106 cells per 0.1 g diaphragm tissue. There were significant differences between mechanical-enzymatic digestion and mechanical grinding method in terms of living cells, inactivated cells, cell aggregates, cell survival rate and suspension concentration (P < 0.05). Moreover, the single cell suspension prepared by the same-volume addition of trypsin and collagenase IV had higher suspension concentration, higher cell survival rate, and less inactivated cells and cell aggregates. To conclude, the single cell suspension of diaphragm tissues could be prepared successful by both mechanical and mechanical-enzyme digestion methods. Mechanical-enzyme digestion is superior to simple mechanical grinding method, with the best single cell suspension after same-volume addition of trypsin, collagenase I and collagenase IV. This is the preferred method for preparation of single cell suspension of the diaphragm tissue.
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BACKGROUND: Many methods have been developed to establish a rabbit VX2 tumor model, but the reliability of each method has not been explored. In order to develop a reliable method, we made some improvements based on the existing methods. OBJECTIVE: To investigate the reliability of rabbit VX2 tumor tissue block implantation and cell suspension via modified and traditional implantation to make the rabbit tibia VX2 tumor model. METHODS: Forty healthy adult New Zealand white rabbits were randomly divided into two groups. Group A was treated with tissue block implantation for tibia VX2 tumor modeling, and group B was treated with cell suspension for tibia VX2 tumor modeling. Modified and traditional implantation was performed on the left and right tibia of the experimental animals, respectively. One hour after successful modeling, ultrasound examination of the puncture site was performed to determine whether there is hematoma. All experimental animals were sacrificed at 3 weeks. X-ray examination of the bilateral tibia was performed to confirm the tumor growth range. Tumor tissue and soft tissue around the puncture site were taken for general and pathological observation to compare the size of the tumor and identify whether there is tumor cell metastasis. RESULTS AND CONCLUSION: One rabbit died in the tissue block group, and all the experimental animals in the cell suspension group survived. X-ray examination indicated the tumors in the tissue block group invaded the cortex, but the tumors in the cell suspension group did not invade the cortex. Gross observation revealed that the tumor volume of the tissue block group was greater than that of the cell suspension group. In the tissue block group, there were one and seven cases of hematoma around the puncture site at 1 hour after modified and traditional implantation, respectively. In the cell suspension group, there were two and nine cases of hematoma around the puncture site at 1 hour after modified and traditional implantation, respectively. Pathological examination showed that local tumor invasion was found in 1 and 8 cases in the tissue block group as well as in 2 and 11 cases in the cell suspension group at 3 months after modified and traditional implantation, respectively. Our findings indicate that the tissue block implantation method is easier and more convenient than the cell suspension method for making rabbit VX2 bone tumors, and the tumor invasion rate of the tissue block implantation method is lower than that of the cell suspension method. Improved tissue block implantation can effectively reduce the tumor invasion rate during modeling.
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Centella asiatica is an important medicinal plant which contains various phytocompounds. Asiatic acid and asiaticosideare two major compounds which are responsible for its various pharmaceutical activities. The present study analyzesthe effect of elicitor, i.e., methyl jasmonate on the synthesis of asiaticoside and asiatic acid (ATA) in shoot, callus, andcell suspension cultures of C. asiatica. A high-performance liquid chromatography analysis showed that the elicitationwith 100 µM concentration of methyl jasmonate enhanced asiaticoside content by 69-fold in callus culture, 39-fold inshoot cultures, and ATA by 1.9-fold in cell suspension culture. Thus, elicitation with methyl jasmonate is an effectivemethod of increasing the rate of biosynthesis of asiaticoside and ATA in plant cell cultures of C. asiatica
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Background: Available options for correction of facial volume loss, such as synthetic fillers, autologous fat and cultured fibroblasts, have limitations viz. temporary effect and high cost. Aim: To assess the use of a novel technique, autologous non-cultured dermal cell suspension transplantation, for correction of localized facial volume loss due to inflammatory pathologies. Methods: It was a pilot study conducted in the Dermatology Outpatient Department, All India Institute of Medical Sciences (AIIMS), New Delhi, India. Autologous non-cultured dermal cell suspension was transplanted in a total of 10 patients, out of which 5 had predominantly dermal loss and the rest had predominantly lipoatrophy. The donor tissue from the gluteal region was digested into a single cell suspension using collagenase-1 and injected into the recipient area. The outcome was assessed subjectively by patients and investigators and objectively using ultrasonography. Cell count, viability testing and measurement of mesenchymal stem cells were also done. Results: On assessment of patients, the median improvement in the predominantly dermal atrophy group at 3 and 6 months was 70% (range: 10–90%) and 80% (range: 0–90%), respectively, and in the predominantly lipoatrophy group, 0% (range: 0–40) and 0% (range: 0–50), respectively. Mean thickness of dermis + subcutis at the baseline was 1.835 mm (range: 0.89–6.04 mm), which increased to 2.912 mm (range: 0.88–7.07 mm, P = 0.03) at 6 months. Limitations: Our pilot study has some limitations such as small sample size and heterogeneity of the recruited patients. Conclusions: Autologous non-cultured dermal cell suspension transplantation appears to be safe and effective in localized facial dermal defects because of inflammatory pathologies, but not effective in deeper defects.
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Objective: To establish the plant tissue culture system of Cistanche tubulosa, and determine the effect of drought stress on accumulation of two respective phenylethanoid glycosides in it. Methods The major chemical constituents of C. tubulosa by tissue culture were analyzed by HPLC-UV and HR-MS. The cell growth curves were also determined. In addition, the effects of drought stress on the phenylethanoid glycosides (echinacoside and acteoside) content in the tissue culture system of C. tubulosa were also studied by using NaCl, mannitol and PEG6000 as osmotic regulators, respectively. Results:Chemical constituents analyses revealed that callus and suspension cultures of C. tubulosa could produce the respective phenylethanoid glycosides of echinacoside and acteoside as in wild plant; Cell growth curves indicated that 30 d were the optimum culture period of callus culture; The cell growth rate and the accumulation of echinacoside and acteoside were mostly inhibited when the callus cells were under drought stress induced by NaCl or mannitol. Meanwhile, the accumulation of echinacoside and acteoside in cell suspension culture of C. tubulosa could be effectively enhanced by treatment with PEG6000. The maximum biomass of echinacoside and acteoside could reach to (1.07 ± 0.10) g/L and (0.12 ± 0.01) g/L 15 d after induction, respectively. And their contents were 20.94% and 2.27% separately based on the cell dry weight (DW) after 15 d of treatment with 6% PEG6000, which were 1.29 and 1.19 fold higher than the control group. Conclusion:Drought stress induced by PEG6000 could effectively enhance the accumulation of echinacoside and acteoside in cell suspension culture of C. tubulosa.
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<p><b>Objective</b>To explore the clinical selection and application of cell suspension examination (CSE) or histopathological technique (HPT) in detecting sperm in the testis tissue obtained by testicular sperm aspiration (TESA) in patients with non-obstructive azoospermia (NOA).</p><p><b>METHODS</b>Totally, 1 006 NOA patients underwent TESA and their testis tissues were subjected to CSE or HPT for sperm detection. Based on the results of CSE, the testicular tissue samples were divided into groups A (with sperm, n = 567) and B (without sperm, n = 439) and the results were compared with those of HPT.</p><p><b>RESULTS</b>HPT showed 508 cases with but 59 without sperm in group A, and 403 with and 36 without sperm in group B. The consistency rate of CSE with that of HPT was 90.56% (Kappa =0.809), and CSE exhibited a significantly higher rate of sperm detection than HPT (56.36% vs 54.08%, P=0.023).</p><p><b>CONCLUSIONS</b>CSE combined with HPT for detecting sperm in the testis tissue of NOA patients undergoing diagnostic TESA helps clinical diagnosis and treatment. The results of CSE have a decisive significance for assisted reproductive therapy, while those of HPT may provide some definite etiological evidence for drug therapy or surgery.</p>
Subject(s)
Humans , Male , Azoospermia , Reproductive Techniques, Assisted , Sperm Retrieval , Spermatozoa , Suspensions , TestisABSTRACT
Vitiligo is a commonly acquired cutaneous depigmentation disorder that affects 0.5~1% of the population worldwide. While phototherapy is the primary treatment for vitiligo, surgical methods can be used for treating patients who are refractory to conventional treatments. Herein, we present the case of a 14-year-old Korean girl who developed vitiligo after hematopoietic stem cell transplantation for the treatment of acute lymphoblastic leukemia. She had multiple depigmented patches on her lower legs that did not respond to narrow-band ultraviolet B phototherapy for 7 months. The depigmented patches were successfully treated by transplantation of non-cultured epidermal cell suspension from the epidermal roof of the suction blister in the right inner thigh. No adverse event, such as secondary infection or scarring in both the donor and recipient sites, was noted. We recommended that non-cultured epidermal cell suspension transplantation using the suction blister method would be a safe and effective option for the treatment of refractory vitiligo.
Subject(s)
Adolescent , Female , Humans , Blister , Cicatrix , Coinfection , Hematopoietic Stem Cell Transplantation , Leg , Methods , Phototherapy , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Suction , Thigh , Tissue Donors , VitiligoABSTRACT
Objective To investigate the Diels-Alder adducts from cell suspension cultures of Morus alba. Methods A variety of column chromatography (CC) including silica gel CC, Sephadex LH-20 CC, C18 CC, and semi-preparative HPLC were used to separate Diels-Alder adducts from cell cultures of M. alba. Their structures were identified by physicochemical properties and various spectroscopic experiments, including MS, NMR, and ECD. Results Eight Diels-Alder adducts were obtained from the ethyl acetate extract of M. alba, and determined as mongolicin H (1), morbilisin J (2), mongolicin F (3), mulberrofuran G (4), artonin D (5), kuwanon R (6), morbilisin C (7), and mulberrofuran E (8). Conclusion Compounds 1-8 are all Diels-Alder adducts and show medium cytotoxic activity, and compounds 1 and 2 are new compounds.
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BACKGROUND: The ABO blood group typing test (ABO test) is an initial pre-transfusion test based on hemagglutination. Although various factors affect hemagglutination strength, few studies have examined how these factors can be applied in clinical laboratories and their effects on hemagglutination. This study was conducted to analyze the factors affecting hemagglutination strength in the ABO test using a tube method applied in many laboratories. METHODS: We conducted a detailed questionnaire survey of 51 laboratories which use the ABO test with a tube method. We also analyzed the results of the ABO test (cell and serum typing) with 40 specimens using factors affecting hemagglutination at a tube method and applied differently in each laboratory. RESULTS: Each laboratory used various methods to prepare red cell suspensions as specimens or reagents and used different reagent to sample ratios, centrifugation protocols, and shaking test tubes before evaluating hemagglutination strength. By testing various combinations of these factors, direct sampling from the red cell layer of the original specimen was found to have the largest effect on lowering hemagglutination strength in cell typing tests. In serum typing tests, various factors influenced hemagglutination strength, including shaking the tube before analysis and the concentration of a home-made red cell suspension used as a reagent. CONCLUSIONS: To achieve accurate results in the ABO test by the tube method, detailed guidelines that include the factors affecting hemagglutination strength determined in this study should be established.
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Centrifugation , Hemagglutination , Indicators and Reagents , Methods , SuspensionsABSTRACT
Cell suspension cultures of Thevetia peruviana were established under dark for 19 days to investigate kinetic behavior related to biomass, substrate, cardiac glycoside, polyphenols, reactive oxygen species and anti-oxidant activity. The results showed high biomass production (18.80gDW/L) as well as sucrose consumption in 7 days. Preferential glucose over fructose consumption was observed. Intracellular production of cardiac glycosides reached 2.58mg DE/gDW at day 19. Highest extracellular production was reached between day 2 and 7 (6.19mg DE/L). Highest extracellular phenolic content was 80.61 ± 5.16mg GAE/L at day 7. Intracellular phenolic content increased to 2.76 ± 0.14mg GAE/gFW at day 7 and remained constant until day 19. ROS production at day 7 could be related to sucrose and glucose total consumption. Pearson Product-Moment Correlation Coefficient (ρ) showed that the phenolic compounds in cell suspension cultures of T. peruviana were responsible for the observed anti-oxidant activity. All together, these results give the first steps in metabolic behavior in cell suspension cultures of T. peruviana.
Se establecieron cultivos en suspensión de la especie vegetal Thevetia peruviana en oscuridad, durante 19 días, para estudiar el comportamiento cinético de producción de biomasa, consumo de sustrato, producción de glicósidos cardiotónicos, polifenoles, especies reactivas de oxígeno y actividad antioxidante. Los resultados mostraron una alta producción de biomasa (18,80g PS/L), al igual que consumo total de sacarosa, a los 7 días de cultivo. Se observó un consumo preferencial de glucosa sobre fructosa durante todo el cultivo. La producción de glicósidos cardiotónicos intracelulares alcanzó valores de 2,58mg ED/g PS, al día 19. La mayor producción extracelular (6,19mg ED/L), se alcanzó entre los días 2 y 7. El mayor contenido de compuestos fenólicos extracelular fue de 80,61 ± 5,16mg GAE/L, en el día 7. El contenido de compuestos fenólicos intracelulares incrementó a 2,76 ± 0,14mg AGE/gPF, al día 7 y se mantuvo constante, hasta el día 19. La producción de EROs, al día 7, puede estar relacionada con el consumo total de sacarosa y glucosa. El coeficiente de correlación producto-momento de Pearson (ρ) indicó que los compuestos fenólicos en cultivos celulares en suspensión de T. peruviana eran los responsables de la actividad antioxidante observada. En conjunto, estos resultados brindan las primeras bases relacionadas al comportamiento metabólico de cultivos celulares en suspensión, de T. peruviana.
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AIM:To investigate the effects of amnion epithelial cell ( AEC) suspension liquid on the biological behavior of the rabbit's corneal epithelium, combined with the in vitro and in vivo experiments. · METHODS: The rabbit's corneal epithelium were cultured in the lower chamber of transwell, and AEC suspension liquid was dropwised in the upper chamber. There was only culture medium in the upper chamber of the control group. The proliferation of rabbit's corneal epithelium was observed with CCK-8 automated colorimetry and the expression of PCNA was detected by immunocytochemistry. We used the scratch wound assay to detect the migration of corneal epithelial cell ( CEC ) . The in vivo models were established by placing a 10mm diameter corneal trephine in the center of the cornea, within 1mol/L NaOH for 1min. We divided those into three groups: treatment group of AEC suspension liquid eye drop, AEC suspension liquid subconjunctival injection and the control group without any treatment. Using the slit- lamp biomicroscope and fluorescence staining to observe the cornea per week. After 28d we took the eyeballs with the HE staining. The expression of VEGF was detected by immunohistochemistry. ·RESULTS: The activity of CEC with AEC treatment was much higher than the control group ( P< 0. 05 ). The expression of PCNA increased in AEC group (P<0. 05). And the migration of CEC in the AEC group was faster than the control one. In vivo, the inflammation of the corneal and the CNV of the AEC group were all significantly reduced compared with the control group (P<0. 05 ). There were less invasive cells and more ordered organization arrangement in ACE group observed by the HE staining. The expression of VEGF and mcp-1 in these two AEC treatment groups all significantly decreased compared with the control group (P<0. 05). ·CONCLUSION: AEC suspension liquid can promote the proliferation and migration of the rabbit's corneal epithelium. The potential of AEC suspension liquid as a therapy for acute corneal alkali burn.
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OBJECTIVE@#To evaluate the impact of plant growth regulators including kinetin (KN), benzyl adenine and naphthalene acetic acid, yeast extract and casein hydrolyzate on biomass accumulation of Vietnamese ginseng Panax vietnamensis (P. vietnamensis) in cell suspension culture.@*METHODS@#Cell suspension cultures were established from friable calluses derived from leaves and petioles of 3-year-old in-vitro P. vietnamensis plants. The cell suspension cultures were grown in Murashige and Skoog basal media supplemented with various concentrations of KN, benzyl adenine, naphthalene acetic acid, and yeast extract and casein hydrolyzate.@*RESULTS@#All tested factors generated an increase in the cell biomass of P. vietnamensis in suspension culture, but the impact of each varies depended on the factor type, concentration, and incubation period. Addition of 2.0 mg/L KN resulted in the largest biomass increase after 24 d, (57.0 ± 0.9) and (3.1 ± 0.1) mg/mL fresh and dry weight, respectively, whereas addition of benzyl adenine or naphthalene acetic acid produced optimum levels of Panax cell biomass at 1.0 and 1.5 mg/L, respectively. Addition of the elicitor yeast extract led to a 1.4-2.4 fold increase in biomass of P. vietnamensis, while addition of casein hydrolyzate enhanced biomass accumulation 1.8-2.6 fold.@*CONCLUSIONS@#The addition of each factor causes significant changes in biomass accumulation of P. vietnamensis. The largest biomass accumulation is from cultures grown in MS media containing 2.0 mg/L KN for 24 d. The outcome of the present study provides new insights into the optimal suspension culture conditions for studies on the in vitro cell biomass production of P. vietnamensis.
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Objective To evaluate the impact of plant growth regulators including kinetin (KN), benzyl adenine and naphthalene acetic acid, yeast extract and casein hydrolyzate on biomass accumulation of Vietnamese ginseng Panax vietnamensis (P. vietnamensis) in cell suspension culture. Methods Cell suspension cultures were established from friable calluses derived from leaves and petioles of 3-year-old in-vitro P. vietnamensis plants. The cell suspension cultures were grown in Murashige and Skoog basal media supplemented with various concentrations of KN, benzyl adenine, naphthalene acetic acid, and yeast extract and casein hydrolyzate. Results All tested factors generated an increase in the cell biomass of P. vietnamensis in suspension culture, but the impact of each varies depended on the factor type, concentration, and incubation period. Addition of 2.0 mg/L KN resulted in the largest biomass increase after 24 d, (57.0 ± 0.9) and (3.1 ± 0.1) mg/mL fresh and dry weight, respectively, whereas addition of benzyl adenine or naphthalene acetic acid produced optimum levels of Panax cell biomass at 1.0 and 1.5 mg/L, respectively. Addition of the elicitor yeast extract led to a 1.4–2.4 fold increase in biomass of P. vietnamensis, while addition of casein hydrolyzate enhanced biomass accumulation 1.8–2.6 fold. Conclusions The addition of each factor causes significant changes in biomass accumulation of P. vietnamensis. The largest biomass accumulation is from cultures grown in MS media containing 2.0 mg/L KN for 24 d. The outcome of the present study provides new insights into the optimal suspension culture conditions for studies on the in vitro cell biomass production of P. vietnamensis.
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Objective Few reports are seen on the methods of establishing the rabbit model of pancreatic cancer .This study was to compare the effect of Panc-1 cell suspension orthotopic implantation with that of VX-2 tissue orthotopic implantation in construc-ting the rabbit model of pancreatic cancer . Methods Using the random number table method , we divided 30 healthy rabbits into a tissue suspension group ( n=15) and a cell suspension group ( n=15) , VX-2 tissue suspension employed for in-situ implanting in the former group and panc-1 cell suspension utilized in the latter .Then we evaluated the two modeling methods by B-ultrasonography , 3.0T MRI, and CT. Results In the third week after modeling , transpla-ntive metastasis of lots of tumor tissues was observed in the duode-num, colon, appendix, and peritoneal wall in 5 rabbits of the tissue suspension group , but only in the greater omentum of 3 rabbits in the cell suspension group , with high signals of MR T 2 in the posterior gastric body .One case of duodenal metastasis was seen in the cell suspension group , with slightly high signals of MR LAVA in the posterior gastric body .The model of pancreatic cancer was successfully established in all the 15 rabbits of the tissue suspension group , but only in 3 of the cell suspension group .The success rate of tumor im-planting at 3 and 4 weeks was significantly higher in the former ( 46.66%and 100%) than in the latter group ( 6.67%and 20.00%) (P<0.05). Conclusion VX-2 tissue orthotopic implantation is a more feasible and convenient method than Panc -1 cell suspension orthotopic implantation for establishing the rabbit model of pancreatic cancer .
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Objective To develop a measurement method for determination of optical parameters of a red blood cell ( RBC) suspension based on the measurement of spatial scattered light signals without using an integrating sphere.Methods Multiple independent photoelectric sensors and light intensity modulation were used to obtain the measured values of diffuse reflectance,diffuse transmittance and collimated transmittance.The measured data results were imported into a Monte Carlo simulation based RTE to inversely determine the absorption coefficient, scattering coefficient and anisotropy factor of the measured sample using a new perturbation method.Results The described measurement method was applied to determine the optical parameters of a polystyrene microsphere suspension with a mean diameter of 2.6μm,and the results were essen-tially consistent with the calculated optical parameters by Mie code.Then, the RBC suspension was used for testing optical parameters,and the results were basically consistent with the parameters in the literature.Conclusion The system based on the measurement of spatial scattered light signals without using an integrating sphere will provide a quick and accurate approach for quantitative analysis of free hemoglobins and RBC suspensions.
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Brazilian flora includes numerous species of medicinal importance that can be used to develop new drugs. Plant tissue culture offers strategies for conservation and use of these species allowing continuous production of plants and bioactive substances.
Subject(s)
Annona/metabolism , Anti-Bacterial Agents/pharmacology , Bacillus thuringiensis/growth & development , Plant Extracts/pharmacology , Plants, Medicinal/metabolism , Streptococcus pyogenes/growth & development , Brazil , Bacillus thuringiensis/drug effects , Cell Culture Techniques , Microbial Sensitivity Tests , Streptococcus pyogenes/drug effectsABSTRACT
Bryophytes were diverse, primitive non vascular amphibious taxa distributed worldwide and form the second largest category of plants. Bryophytes synthesize an array of phytochemicals to combat against the unhospitable environmental conditions including predation, UV radiation, high temperature and pest and pathogens. The present investigation was undertaken to elucidate flavonoids from in vitro cell cultures of the liverwort Marchantia linearis Lehm. & Lindenb. its fractionation and analysis of insecticidal potentialities. Initially, callus culture was initiated from spores in MS/5 media containing growth regulators BAP and NAA at the concentration of 2 mg/L and 0.5 mg/L. Agitation of the friable callus at lower rpm bring about lower level of cell dispersion, on the contrary at higher rpm might have risk of cell collision that is why rpm was kept at moderate speed i.e., 110 rpm. Continuous sub culturing process substantially improves cell growth and biomass. In the second phase, the flavonoids were isolated from cell suspension cultures of M. linearis and were fractionated by TLC and HPLC PAD chromatogram, which revealed the presence of quercetin, luteolin, apigenin, rutin and kaempferol. In vivo insecticidal analysis revealed significant antifeedant, larvicidal and pupicidal activities at all the concentrations against 5th instar larvae of Spodoptera litura. The extract also exhibited feeding deterrent activity with M. linearis. Similarly, the nutritional parameters were also affected i.e., reduced ECI (Efficiency of conversion of ingested food) and ECD (Efficiency of conversion of digested food) and increased AD (Approximate digestibility) and metabolic cost for the larvae, when compared with the control. The consumption of the basal diet with the incorporation of flavonoids by S. litura larvae was not significantly different compared to the consumption of the control diet by the larvae. Faecal production reduced proportionally with concentrations of the extract.Bryophytes were diverse, primitive non vascular amphibious taxa distributed worldwide and form the second largest category of plants. Bryophytes synthesize an array of phytochemicals to combat against the unhospitable environmental conditions including predation, UV radiation, high temperature and pest and pathogens. The present investigation was undertaken to elucidate flavonoids from in vitro cell cultures of the liverwort Marchantia linearis Lehm. & Lindenb. its fractionation and analysis of insecticidal potentialities. Initially, callus culture was initiated from spores in MS/5 media containing growth regulators BAP and NAA at the concentration of 2 mg/L and 0.5 mg/L. Agitation of the friable callus at lower rpm bring about lower level of cell dispersion, on the contrary at higher rpm might have risk of cell collision that is why rpm was kept at moderate speed i.e., 110 rpm. Continuous sub culturing process substantially improves cell growth and biomass. In the second phase, the flavonoids were isolated from cell suspension cultures of M. linearis and were fractionated by TLC and HPLC PAD chromatogram, which revealed the presence of quercetin, luteolin, apigenin, rutin and kaempferol. In vivo insecticidal analysis revealed significant antifeedant, larvicidal and pupicidal activities at all the concentrations against 5th instar larvae of Spodoptera litura. The extract also exhibited feeding deterrent activity with M. linearis. Similarly, the nutritional parameters were also affected i.e., reduced ECI (Efficiency of conversion of ingested food) and ECD (Efficiency of conversion of digested food) and increased AD (Approximate digestibility) and metabolic cost for the larvae, when compared with the control. The consumption of the basal diet with the incorporation of flavonoids by S. litura larvae was not significantly different compared to the consumption of the control diet by the larvae. Faecal production reduced proportionally with concentrations of the extract.