Visible Light and Its Influence on the Embryonic Viability of the Cricket Acheta domesticus

During in vitro fertilization, human embryos are incubated without light, and these conditions do not ensure embryo survival. This study explored whether environmental conditions can influence the embryo viability rates of the house cricket, Acheta domesticus. In particular, the experiment tested what colors of visible light provide the best incubation conditions to ensure cricket embryo viability. The concept was to use house cricket embryos to represent human embryos. Cricket embryos were chosen as their eggs have soft outer membrane casings and resemble human embryos during the first few days after fertilization. During the experiment, the adult crickets laid their eggs into one of six soil-filled boxes called substrates. Each substrate was placed into one of six storage containers filled with adult crickets and lit with a different colored visible light (red, yellow, green, blue, white, or no light). After two days of breeding, the egg-filled substrates were removed from the adult crickets and placed in another storage container of the same color light. After incubation under heat-emitting lamps and under one of six light colors, nymphs were counted after hatching to determine embryo viability. After three trials, the red light provided the significantly highest viability rate, with yellow and no light being comparable seconds. The green, blue, and white lights showed significantly lower viability rates than no visible light. My results raise the speculation that exposing fertilized mammal eggs to visible light colors might have the same effects during the in vitro fertilization process.

Viability and Action of CPL Lectin on <i>in Vitro</i>Germinability of Pollen Grains of <i>Malpighia emarginata</i>DC.—(Malpighiaceae)

This study aimed to observe the viability of the pollen grains of Malpighia emarginata DC. (West Indian cherry) and the action of a lectin in their germination. Lectins are proteins that specifically interact with carbohydrates, but don’t modify them and bind with high affinity and specificity, promoting a transfer of information that is clearly central to many cellular processes in living beings. For the viability test was used aniline blue in lactophenol. The in vitro germination test used was the hanging drop method, in control medium containing sucrose, boric acid, calcium nitrate and agar. The experiment dealt with three different treatments in order to pollen germination: growing medium without addition of lectin, with 1.0 μg/ml and with 3.0 μg/ml CPL lectin. This lectin is extracted from seeds of Crotalaria pallida L.-Leguminosae. Data on pollen grains were analyzed by one-way ANOVA, with 95% reliability and comparison of averages by Turkey test at 5% probability. It was found that despite previous high viability of the pollen grains (c. 70%), a germinability rate has been low, yet exceptionally larger in pollen samples placed on a growing medium with addition of 3.0 μg/ml of CPL lectin (24%). There was no significant difference among the pollen grains germinate in medium without lectin and those in medium with the addition of only 1.0 μg/ml. According to data obtained in the treatments, CPL lectin, with concentration of 3.0 μg/ml, influenced the formation of the pollen tube and thus more pollen germinated in Malpighia emarginata.

A review on cell damage,viability,and functionality during 3D bioprinting

Three-dimensional(3D)bioprinting fabricates 3D functional tissues/organs by accurately depositing the bioink composed of the biological materials and living cells.Even though 3D bioprinting techniques have experienced significant advancement over the past decades,it remains challenging for 3D bioprinting to artificially fabricate functional tissues/organs with high post-printing cell viability and functionality since cells endure various types of stress during the bioprinting process.Generally,cell viability which is affected by several factors including the stress and the environmental factors,such as pH and temperature,is mainly determined by the magnitude and duration of the stress imposed on the cells with poorer cell viability under a higher stress and a longer duration condition.The maintenance of high cell viability especially for those vulnerable cells,such as stem cells which are more sensitive to multiple stresses,is a key initial step to ensure the functionality of the artificial tissues/organs.In addition,maintaining the pluripotency of the cells such as proliferation and differentiation abilities is also essential for the 3D-bioprinted tissues/organs to be similar to native tissues/organs.This review discusses various pathways triggering cell damage and the major factors affecting cell viability during different bioprinting processes,summarizes the studies on cell viabilities and functionalities in different bioprinting processes,and presents several potential approaches to protect cells from injuries to ensure high cell viability and functionality.

Effect of different drying methods on the amino acids,α-dicarbonyls and volatile compounds of rape bee pollen

The significant demand for high quality food has motivated us to adopt appropriate processing methods to improve the food nutritional quality and flavors.In this study,the effects of five drying methods,namely,pulsed vacuum drying(PVD),freeze drying(FD),infrared drying(IRD),hot-air drying(HAD)and sun drying(SD)on free amino acids(FAAs),α-dicarbonyl compounds(α-DCs)and volatile compounds(VOCs)in rape bee pollen(RBP)were determined.The results showed that FD significantly released the essential amino acids(EAAs)compared with fresh samples while SD caused the highest loss.Glucosone was the dominantα-DCs in RBP and the highest loss was observed after PVD.Aldehydes were the dominant volatiles of RBP and SD samples contained more new volatile substances(especially aldehydes)than the other four drying methods.Comprehensively,FD and PVD would be potential methods to effectively reduce the quality deterioration of RBP in the drying process.

Viability and hatchability of brine shrimp Artemia franciscana cysts after passing through the digestive system of eared grebes Podiceps nigricollis

Brine shrimp Artemia franciscana provide food for many migrating and staging birds that spend summer and fall on Great Salt Lake,Utah,USA.Artemia produce live young and cysts(hard-walled eggs);these cysts are commercially harvested on Great Salt Lake and support a large industry in Utah.It is unclear the impact that millions of hungry birds have on the Artemia population in the lake.To help assess that,this study evaluated cyst viability(percentage of cysts that contain an embryo)and hatchability(percent of cysts that hatch)from cysts that had passed through the digestive tract of eared grebes Podiceps nigricollis and cysts obtained directly from Great Salt Lake at the same site where each grebe was collected.Hatchability was significantly higher for cysts collected from the water column(19%)than from the stomach(0.3%)or intestines(3%)of eared grebes.Viability also was significantly different for cysts collected from the water column(29%),stomach(0.7%),and intestines(5%).These results indicate that eared grebes nutritionally benefit from eating cysts and that they may be an important food source for grebes in late fall after the adult population of Artemia dies off due to the water becoming too cold.Also,enough cysts survive their passage through the digestive system that grebes can vector hatchable cysts to other waterbodies.

Pollen viability of Polygala paniculata L.(Polygalaceae)using different staining methods

Polygala paniculata L.is a medicinal plant that grows in the Brazilian Atlantic coast,known as‘barba-de-São-João’,‘barba-de-bode’,‘vassourinha branca’,and‘mimosa’.In this study,pollen viability was estimated by three different staining methods:2%acetic orcein,2%acetic carmine,and Alexander’s stain.The young inflorescences of twenty accessions were collected and fixed in a solution of ethanol:acetic acid(3:1)for 24 hours,then stored in ethanol 70%under refrigeration.Six slides per plant,two for each stain,were prepared by squashing,and 300 pollen grains per slide were analyzed.Pollen viability was high(>70%)for most accessions of P.paniculata using the Alexander’s stain,which proved the most adequate method to estimate pollen viability.

Modern pollen-vegetation relationships along altitudinal vegetation belts in the Leigong Mountain of southwestern China

Understanding the distribution,dispersal,and correlation of modern pollen with vegetation in mountainous regions is essential for establishing accurate modern analogs for fossil pollen records.This study,conducted in Leigong Mountain on the YunnanGuizhou Plateau of southwestern China,involved the collection of 35 surface soil samples from diverse vegetation communities along an elevational gradient ranging from 1210 to 1875 meters.The results reveal a close correspondence between modern pollen assemblages and vegetation zones.Principal Component Analysis(PCA)results indicate that pollen assemblages can effectively distinguish between subtropical montane evergreen broad-leaved forest(SEBF)and subtropical montane deciduous broadleaved forest(SDBF).However,both SEBF and SDBF show significant overlap with subtropical montane evergreen-deciduous broad-leaved mixed forest(SEMF).Detrended Correspondence Analysis(DCA)results clearly distinguish the three vegetation zones,and the first axis of DCA shows a significant positive correlation with elevation(p<0.01,R=0.48).Discriminant Analysis(DA)successfully assigns 94.4%of the modern pollen samples to their respective vegetation zones.Pollen taxa such as Impatiens,Astertype,and Rosaceae exhibit significant indicative capabilities for the SEBF zone,effectively distinguishing this vegetation zone from others.Pinus and Alnus display overrepresentation in the Leigong Mountain region,while Quercus(D,deciduous-type)and Poaceae exhibit high representation in the SEBF zone.In the SEBF zone,both pollen diversity and richness are the lowest.Our study reveals the complex relationship between the richness and diversity of pollen and vegetation.The diversity and richness of tree and shrub pollen are found to be lower than those of the corresponding plants.The pollen-vegetation relationship elucidated in this study serves as a critical reference for reconstructing ancient environments from fossil pollen retrieved in this region.

High temperature treatment induced production of unreduced 2n pollen in Camellia oleifera

Unreduced gametes through chromosome doubling play a major role in the process of plant polyploidization.Our previous work confirmed that Camellia oleifera can produce natural 2n pollen,and it is possible to induce the 2n pollen formation by high temperature treatment.This study focused on the optimization of the 2n pollen induction technique and the mechanisms of high temperature-induced2n pollen formation in C.oleifera.We found that the optimal protocol for inducing 2n pollen via high temperature was to perform 45℃with4 h at the prophaseⅠstage of the pollen mother cells(PMCs).Meanwhile,high temperature significantly decreased the yield and fertility of2n pollen.Through the observation of meiosis,abnormal chromosome and cytological behaviour was discovered under high-temperature treatment,and we confirmed that the formation of 2n pollen is caused by abnormal cell plate.Based on weighted gene co-expression network analysis,fifteen hub genes related to cell cycle control were identified.After male flower buds were exposed to heat shock,polygalacturonase gene(CoPGX3)was significantly upregulated.We inferred that high temperature causes the CoPGX3 gene to be overexpressed and that CoPGX3 is redistributed into the cytosol where it degrades cytoplasmic pectin,which leads to an abnormal cell plate.Furthermore,abnormal cytokinesis resulted in the formation of dyads and triads,and PMCs divided to produce 2n pollen.Our findings provide new insights into the mechanism of 2n pollen induced by high temperature in a woody plant and lay a foundation for further ploidy breeding of C.oleifera.

Decreased Pollen Viability and Thicken Pollen Intine in Antisense Silenced Brassica campestris Mutant of BcMF19

Brassica campestris male fertility 19 （BcMF19; GenBank accession number GQ902048.1）, a gene that is specially expressed in tapetum cells and microspores during anther development in B. campestris ssp. chinensis, which is learned from the previous in situ hybridization study. In the present study, we constructed antisense-silenced plants of BcMF19 using B. campestris ssp. chinensis to validate this prediction. The morphology of the pistils, long anthers, and short anthers was significantly affected in 35sbcmf19 compared with the control samples. 4＇-6-Diamidino-2-phenylindole staining revealed that two generative nuclei and one large vegetative nucleus were not affected in the mutant compared with control. Statistical analysis of Alexander＇s staining results showed that 96% of the control pollen grains had vitality, whereas only 86% of the mutant pollen grains did. Under scanning electron microscopy, the mutant demonstrated numerous abnormal pollen grains and resembled dried persimmon. The frequency of normal pollen grains was approximately 18%. Under transmission electron microscopy, the pollen intine during the binucleate and mature pollen stages in 35sbcmf19 exhibited abnormal thickening, especially at the germinal furrows, compared with control. In vitro pollen germination test showed that the tips of the mutant pollen tubes transformed into globular alveoli and stopped growing compared with control. On the other hand, in vivo pollen germination test suggested that BcMF19 affected the pollen tube extension in the pistil. These findings indicate that BcMF19 is essential to the pollen development and pollen tube extension orB. campestris ssp. chinensis.

GAMYB transcription factor LoMYB65 from lily plays a vital role in pollen development

Lily(Lilium spp.)is an important horticultural crop,but its use is limited due to serious pollen contamination problems.There are many studies on pollen development in model plants,but few on flower crops such as lilies.Gibberellin(GA)is a large class of hormones and plays an important role in plant vegetative growth and reproductive development.GAMYB is a group of the R2R3-MYB family upregulated by gibberellin,and plays an important role in anther development.Here,we isolated a novel GAMYB,named LoMYB65,from lily,which was closely related to the AtMYB65 and AtMYB33 in Arabidopsis.Fluorescence quantitative PCR results showed that LoMYB65 was mainly expressed in lily anthers.LoMYB65 could be activated by 288μmol·L^(-1)GA3treatment and the LoMYB65 protein was located in the nucleus and cytoplasm,and had transactivation in yeast and tobacco leaf cells.The conserved motif within 226 amino acids of the C-terminal of LoMYB65 contributed to its transactivation.Overexpression of LoMYB65 caused dwarf phenotype,unnormal tapetum development,less seeds of siliques in transgenic Arabidopsis plants,the transgenic plants showed partly male sterile.Simultaneously,silencing of LoMYB65 with VIGS(Virus Induced Gene Silencing)in lily anthers caused unnormal pollen development and reduced the pollen amount.Overexpression of LoMYB65 in Arabidopsis and silencing of LoMYB65 in lily resulted in decreased pollen counts,so we speculate that LoMYB65 may be dose-dependent.Overall,these findings suggest that LoMYB65 may play an important role in anther development and pollen formation in lily.LoMYB65 may provide a useful candidate gene for pollenless breeding of lily.

Effect of wall-disruption on nutrient composition and in vitro digestion of camellia and lotus bee pollens

The nutrient digestion,absorption and biological activity of bee pollen may be limited due to the complex pollen wall.Here,the effect of superfine grinding technology on the release of nutrients from bee pollen were investigated,and their antioxidant activities and in vitro digestion were explored in this study.Results showed that the content of nutrients in bee pollen increased after wall disruption.Among them,fat content increased by 22.55%-8.31%,protein content increased by 0.54%-4.91%,starch content increased by 36.31%-48.64%,soluble sugar content increased by 20.57%-29.67%,total phenolic acid content increased by 11.73%-86.98%and total flavonoids content increased by 14.29%-24.79%.At the same time,the antioxidant activity increased by 14.84%-46.00%.Furthermore,the active components such as phenolic compounds in the wall-disruption bee pollen were more readily to be released during the in vitro digestion,and easier to be absorbed because of their higher bioaccessibility.Antioxidant activities during in vitro digestion were also improved in walldisruption bee pollen.These findings provide evidence that bee pollen wall disruption was suggested,thus,it is more conducive to exerting the value of bee pollen in functional foods.

Allergen degradation of bee pollen by lactic acid bacteria fermentation and its alleviatory effects on allergic reactions in BALB/c mice

Food allergy as a global health problem threatens food industry.Bee pollen(BP)is a typical food with allergenic potentials,although it performs various nutritional/pharmacological functions to humans.In this study,lactic acid bacteria(LAB)were used to ferment Brassica napus BP for alleviating its allergenicity.Four novel allergens(glutaredoxin,oleosin-B2,catalase and lipase)were identified with significant decreases in LAB-fermented BP(FBP)than natural BP by proteomics.Meanwhile,metabolomics analysis showed significant increases of 28 characteristic oligopeptides and amino acids in FBP versus BP,indicating the degradation of LAB on allergens.Moreover,FBP showed alleviatory effects in BALB/c mice,which relieved pathological symptoms and lowered production of allergic mediators.Microbial high-throughput sequencing analysis showed that FBP could regulate gut microbiota and metabolism to strengthen immunity,which were closely correlated with the alleviation of allergic reactivity.These findings could contribute to the development and utilization of hypoallergenic BP products.

DIA-based proteome profiling with PRM verification reveals the involvement of ER-associated protein processing in pollen abortion in Ogura CMS cabbage

Ogura cytoplasmic male sterility(Ogura CMS)is extensively applied in hybrid seed production in cruciferous crops.However,the posttranscriptional molecular basis of Ogura CMS in cruciferous crops remains elusive.Here,a data-independent acquisition-based proteomic approach coupled with a parallel reaction monitoring-based targeted proteomic assay was used to analyze the proteome dynamics of Ogura CMS cabbage line RM and its maintainer line RF during floral bud development to obtain insights into the mechanism underlying Ogura CMS in cruciferous crops.A total of 9162 proteins corresponding to 61464 peptides were identified in RM and RF floral buds.The proteomic fluctuation of RM was weaker than that of RF.Differences in protein expression between RM and RF gradually enlarged with floral bud development.Fifteen continually up-regulated and eight continually down-regulated proteins were found in RM relative to RF throughout floral bud development.Differentially expressed proteins between RM and RF during floral bud development were implicated in the endoplasmic reticulum(ER)-associated protein processing pathway,in which most of them exhibited down-regulated expression in RM.These data suggest that ER-associated protein processing may be involved in pollen abortion in Ogura CMS cabbage by inhibiting the expression of critical factors.Our findings not only deepen the understanding of the molecular mechanisms of Ogura CMS in cruciferous crops but also provide better guidance for applying Ogura CMS in the hybrid breeding of cruciferous crops.

甲醛固定对Live/Dead BacLight Bacterial Viability Kit死活细菌染液荧光显微计数海洋细菌的影响

利用Live/Dead BacLight Bacterial Viability Kit死活细菌染液对采自湛江东海大堤海水、沉积物细菌和大型海藻拟刚毛藻(Cladophoropsis zollingeri)内生细菌数量进行了甲醛固定处理前后的荧光显微计数对比分析。结果表明,新鲜样品(不加甲醛固定)、甲醛刚固定样品、甲醛固定1周样品和甲醛固定2周样品中海洋细菌数量差异不显著(p>0.05)。甲醛固定对Live/Dead BacLight Bacterial Viability Kit死活细菌染液荧光显微计数海洋细菌数量无显著影响,固定后的样品可在2周内完成计数。

Determination of A canthopanax senticosus ＇ s Seed Viability

以6个产地的刺五加种子为试验材料，采用四唑染色法（TTC法）、红墨水染色法、溴麝香草酚蓝法（BTB法）、炒种法及纸上荧光法测定种子生活力，筛选最佳的刺五加种子生活力测定方法。结果显示：溴麝香草酚蓝法是测定刺五加种子生活力的最佳方法。

Effects of primary PCI and facilitated PCI on myocardial viability and ventricular systolic synchrony in acute myocardial infarction patients

Objective To evaluate short time effects of primary percutaneous coronary intervention (pPCI) and rtPA thrombolysis+PCI (rtPA+PCI) on myocardial viability and ventricular systolic synchrony in AMI patients.Methods Eighty seven patients with first AMI were divided into two groups: group A ( n =42), pPCI group, the patients underwent PCI within 6h after onset of AMI; group B ( n =45), rtPA+PCI group, the patients underwent PCI after thrombolysis within 6h after onset of AMI; Myocardial viability was measured by 99m Tc MIBI SPECT. While, the parameters of cardiac function LVEF and ventricular systolic synchrony LVPS were measured by 99m Tc gated cardiac blood pool image on the first and the fourth weekend. Results (1) The peak CK MB was significantly lower in group A than that in group B( P <0.01 ). (2) Myocardial infarction area (MIA) was decreased and radioactivity counts in MIA was significantly increased in group A and B on the 4th weekend compared with that on the first weekend ( P <0.01 ), but there were no significant difference between group A and group B. (3) LVEF, LVPS were no significant difference between group A and group B.Conclusions (1)pPCI in acute myocardial infartion can limit infarct area, maintain ventricular systolic synchrony and improve ventricular function; (2) but, in those hospitals that there were no any condition for PCI, they should transfer the patients to central hospital for PCI after thrombolysis at the first time. It is beneficial to improve myocardial viability and ventricular systolic synchrony of AMI patients in short time.

Influence of perfusate on liver viability during hypothermic machine perfusion

AIM: To optimize the perfusates used for hypothermicmachine perfusion(HMP).METHODS: Sprague-Dawley rats were assigned randomly to three groups(n = 12 per group) that received either saline, University of Wisconsin coldstorage solution(UW) or histidine-tryptophan-ketoglutarate solution(HTK) as the perfusate. Each group was divided into two subgroups: static cold storage(SCS) and HMP(n = 6 per subgroup). The liver graft was retrieved according to the method described by Kamada. For the SCS group, the graft was directly placed into cold perfusate(0-4?℃) for 6 h after liver isolation while the portal vein of the graft was connected to the perfusion machine for the HMP group. Then the perfusates were collected at different time points for analysis of aspartate aminotransferase(AST), alanine transaminase(ALT) and lactate dehydrogenase(LDH) levels. Liver tissues were obtained for evaluation of histology, dry/wet weight(D/W) ratio, and malondialdehyde(MDA) and adenosine-triphosphate(ATP) levels. The portal vein pressure and velocity were monitored in real time in all HMP subgroups.RESULTS: Comparison of HMP and SCS: Regardless of the perfusate, HMP improved the architecture of donor graft in reducing the congestion around sinusoids and central vein and maintaining sinusoid lining in morphology; HMP improved liver function in terms of ALT, AST and LDH, especially during the 3-6 h period(SCS vs HMP using saline: ALT3, 225.00 ± 105.62 vs 49.50 ± 18.50, P = 0.047; LDH3, 1362.17 ± 563.30 vs 325.75 ± 147.43, P = 0.041; UW: LDH6, 2880.14 ± 948.46 vs 2135.00 ± 174.27, P = 0.049; HTK, AST6, 307.50 ± 52.95 vs 185.20 ± 20.46, P = 0.041); HMP decreased MDA level(saline, 2.79 ± 0.30 vs 1.09 ± 0.09, P = 0.008; UW, 3.01 ± 0.77 vs 1.23 ± 0.68, P = 0.005; HTK, 3.30 ± 0.52 vs 1.56 ± 0.22, P = 0.006). Comparison among HMP subgroups: HTK showed less portal vein resistance than UW and saline(vs saline, 3.41 ± 0.49 vs 5.00 ± 0.38, P < 0.001; vs UW, 3.41 ± 0.49 vs 4.52 ± 0.63, P = 0.007); UW reduced edema most efficiently(vs saline, 0.68 ± 0.02 vs 0.79 ± 0.05, P = 0.013), while HTK maintained ATP levels best(vs saline, 622.60 ± 29.11 vs 327.43 ± 44.66, P < 0.001; vs UW, 622.60 ± 29.11 vs 301.80 ± 37.68, P < 0.001).CONCLUSION: HMP is superior to SCS in maintaining both architecture and function of liver grafts. Further, HTK was found to be the optimal perfusate for HMP.

Protective Effects of Trimetazidine on Bone Marrow Mesenchymal Stem Cells Viability in an ex vivo Model of Hypoxia and in vivo Model of Locally Myocardial Ischemia

Bone marrow mesenchymal stem cells (MSCs) have shown potential for cardiac repair following myocardial injury,but this approach is limited by their poor viability after transplantation.The present study was to investigate whether trimetazidine (TMZ) could improve survival of MSCs in an ex vitro model of hypoxia,as well as survival,differentiation,and subsequent activities of transplanted MSCs in rat hearts with acute myocardial infarction (AMI).MSCs at passage 3 were examined for their viability and apoptosis under a transmission electron microscope,and by using flow cytometry following culture in serumfree medium and exposure to hypoxia (5% CO2,95% N2) for 12 h with or without TMZ.Thirty Wistar rats were divided into 3 groups (n=10 each group),including groupⅠ(AMI control),groupⅡ (MSCs transplantation alone),and group Ⅲ (TMZ+MSCs).Rat MSCs (4×107) were injected into peri-infarct myocardium (MSCs group and TMZ+MSCs group) 30 min after coronary artery ligation.The rats in TMZ+MSCs group were additionally fed on TMZ (2.08 mg?kg-1?day-1) from day 3 before AMI to day 28 after AMI.Cardiac structure and function were assessed by echocardiography at 28th day after transplantation.Blood samples were collected before the start of TMZ therapy (baseline),and 24 and 48 h after AMI,and inflammatory cytokines (CRP,TNF-α) were measured.Then the sur-vival and differentiation of transplanted cells in vivo were detected by immunofluorescent staining.The cellular apoptosis in the peri-infarct region was detected by using TUNEL assay.Furthermore,apoptosis-related proteins (Bcl-2,Bax) within the post-infarcted myocardium were detected by using Western blotting.In hypoxic culture,the TMZ-treated MSCs displayed a two-fold decrease in apoptosis under serumfree medium and hypoxia environment.In vivo,cardiac infarct size was significantly reduced,and cardiac function significantly improved in MSCs and TMZ+MSCs groups as compared with those in the AMI control group.Combined treatment of TMZ with MSCs implantation demonstrated further decreased MSCs apoptosis,further increased MSCs viability,further decreased infarct size,and further improved cardiac function as compared with MSCs alone.The baseline levels of inflammatory cyto-kines (CRP,TNF-α) had no significant difference among the groups.In contrast,all parameters at 24 h were lower in TMZ+MSCs group than those in MSCs group.Furthermore,Western blotting indicated that the expression of antiapoptotic protein Bcl-2 was upregulated,while the proapoptotic protein Bax was down-regulated in the TMZ+MSCs group,compared with that in the MSCs group.It is suggested that implantation of MSCs combined with TMZ treatment is superior to MSCs monotherapy for MSCs viability and cardiac function recovery.

Effect of human autologous serum and fetal bovine serum on human corneal epithelial cell viability,migration and proliferation in vitro

AIM： To analyze the concentration-dependent effects of autologous serum （AS） and fetal bovine serum （FBS） on human corneal epithelial cell （HCEC） viability, migration and proliferation. METHODS： AS was prepared from 13 patients with non- healing epithelial defects Dulbecco＇s modified eagle medium/ Ham＇s F12 （DMEM/F12） with 5% FBS, 0.5% dimethyl sulphoxide （DMSO）, 10 ng/mL human epidermal growth factor, 1% insulin-transferrin-selenium, then were incubated in serum media： DMEM/F12 supplemented by 5%, 10%, 15% or 30% AS or FBS. HCEC viability was analyzed using cell proliferation kit XTI＇, migration using a wound healing assay, proliferation by the cell proliferation enzyme-linked immunosorbent assay （ELISA） BrdU kit. Statistical analysis was performed using the generalized linear model, the values at 30% AS or 30% FBS were used as the baselines. RESULTS： HCEC viability was the highest at 30% AS or 15% FBS and the lowest at 10% AS or 30% FBS application. HCEC migration was the quickest through 30% AS or 30% FBS and the slowest through 5% AS or 5% FBS concentrations. Proliferation was the most increased through 15% AS or 5% FBS and the least increased through 30% AS or 30% FBS concentrations. HCEC viability at 10% and 15% AS was significantly worse （P=0.001, P=0.023） compared to baseline and significantly better at 15% FBS （P=0.003） concentrations. HCEC migration was significantly worse （P〈0.007） and HCEC proliferation significantly better （P〈0.001） in all concentration groups compared to baseline. CONCLUSION： For the best viability of HCEC 30% AS or 15% FBS, for HCEC migration 30% AS or 30% FBS, for proliferation 15% AS or 5% FBS should be used. Therefore, we suggest the use of 30% AS in clinical practice.