Effect Study of the Recombinant Human Brain Natriuretic Peptide in Patients with Heart Failure Combined with Hypotension

Objective: This paper aims to investigate the effect of applying recombinant human brain natriuretic peptide in patients with heart failure combined with hypotension. Recombinant human brain natriuretic peptide is a synthetic polypeptide drug that is primarily used to treat acute heart failure. Its mechanism of action closely mimics that of human endogenous brain natriuretic peptide. By binding to receptors on cardiomyocytes, it exerts its pharmacological effects. Methods: For the study, 76 heart failure patients with hypotension were selected from our hospital between May 2022 and June 2023. These patients were divided into two groups: a control group and an observation group, each comprising 38 patients. The control group received dopamine treatment, while the observation group was treated with recombinant brain natriuretic peptide. The objective was to compare the effects of the treatments in both groups by analyzing cardiac function indices and levels of vasoactive substances to identify any significant differences in outcomes. Results: The overall response rate of the patients in the observation group and the control group was 94.74% and 73.68%, significantly higher as compared with the observation group (P 0.05). After the following treatment, BNP, ANNP and urine output in the observation group were significantly different compared with the control group, of the statistical significance (P Conclusion: For the treatment of heart failure patients with hypotension, the clinical application of recombinant human brain natriuretic peptide is the most ideal, and significantly improves the cardiac function of patients, which is worth popularizing.

Lyophilized recombinant human brain natriuretic peptide for chronic heart failure:Effects on cardiac function and inflammation

BACKGROUND Chronic heart failure(CHF)is a serious and prevalent condition characterized by impaired cardiac function and inflammation.Standard therapy for CHF has limitations,prompting the exploration of alternative treatments.Recombinant human brain natriuretic peptide(BNP)has emerged as a potential therapy,with evidence suggesting that it can improve cardiac function and reduce inflammation in patients with CHF.However,further research is required to determine the efficacy and safety of lyophilized recombinant human BNP in CHF patients and its impact on microinflammatory status.This study aimed to investigate the effects of lyophilized recombinant human BNP therapy on CHF patients’cardiac function and microinflammatory status.AIM To investigate the effects of freeze-dried recombinant human BNP therapy on cardiac function and microinflammatory status in patients with CHF.METHODS In total,102 CHF patients admitted to our hospital from January 2021 to January 2022 were randomly assigned to control and observation groups(n=51 patients/group).The control patients were treated with standard HF therapy for 3 d,whereas the observational patients were injected with the recombinant human BNP for 3 d.Clinical efficacy,inflammatory factor levels,myocardial damage,cardiac function before and after the treatment,and adverse reactions during treatment were compared between the two groups.RESULTS The overall clinical efficacy was higher in the observation group than in the control group.Compared with baseline,serum hypersensitive C-reactive protein,N-terminal proBNP,and troponin I level,and physical,emotional,social,and economic scores were lower in both groups after treatment,with greater reductions in levels and scores noted in the observation group than in the control group.The overall incidence of adverse reactions in the observation group was not significantly different compared with that in the control group(P>0.05).CONCLUSION Freeze-dried recombinant human BNP therapy can improve heart function and enhance microinflammatory status,thereby improving overall quality of life without any obvious side effects.This therapy is safe and reliable.

Photon penetration depth in human brain for light stimulation and treatment:A realistic Monte Carlo simulation study

Light has been clinically utilized as a stimulation in medical treatment,such as Low-level laser therapy and photodynamic thenapy,which has been more and more widely accepted in public.The penetration depth of the treatment light is important for precision treatment and safety control.The isue of light penetration has been highlighted in biomedical optics field for decades.However,quantitative research is sparse and even there are conflicts of view on the capability of near-infrared light penet ration into brain tissue.This study attempts to quantitatively revisit this issue by innovative high-realistic 3D Monte Caro modeling of stimulated light penetnation within high precision Visible Chinese human head.The properties of light,such as its wave-length,ilumination profle and size are concern in this study.We made stra ightforward and quantitative comparisons among the ffects by the light properties(i.e.,wavelengths:660,810 and 980 nn;beam types:Gaussi an and flat beamn;bear dianeters:0,2,4 and 6 cm)which are in the range of light treatment.The findings include about 3%of light dosage within brain tisue;the combination of Gaussian beam and 810nm light make the max imum light penetration(>5cm),which allows light to cross through gray matter into white mater.This study offered us,the first time as we know,quantitative guide for light stimulation parameter optimization in medical treatment.

Are human dental papilla-derived stem cell and human brain-derived neural stem cell transplantations suitable for treatment of Parkinson’s disease?

Transplantation of neural stem cells has been reported as a possible approach for replacing impaired dopaminergic neurons. In this study, we tested the efficacy of early-stage human dental papilla-derived stem cells and human brain-derived neural stem cells in rat models of 6-hydroxydopamine-induced Parkinson＇s disease. Rats received a unilateral injection of 6-hydroxydopamine into right medial forebrain bundle, followed 3 weeks later by injections of PBS, early-stage human dental papilla-derived stem cells, or human brain-derived neural stem cells into the ipsilateral striatum. All of the rats in the human dental papilla-derived stem cell group died from tumor formation at around 2 weeks following cell transplantation. Postmortem examinations revealed homogeneous malignant tumors in the striatum of the human dental papilla-derived stem cell group. Stepping tests revealed that human brain-derived neural stem cell transplantation did not improve motor dysfunction. In apomorphine-induced rotation tests, neither the human brain-derived neural stem cell group nor the control groups （PBS injection） demonstrated significant changes. Glucose metabolism in the lesioned side of striatum was reduced by human brain-derived neural stem cell transplantation. [18F]-FP-CIT PET scans in the striatum did not demonstrate a significant increase in the human brain-derived neural stem cell group. Tyrosine hydroxylase （dopaminergic neuronal marker） staining and G protein-activated inward rectifier potassium channel 2 （A9 dopaminergic neuronal marker） were positive in the lesioned side of striatum in the human brain-derived neural stem cell group. The use of early-stage human dental papilla-derived stern cells confirmed its tendency to form tumors. Human brain-derived neural stem cells could be partially differentiated into dopaminergic neurons, but they did not secrete dopamine.

Differential Spontaneous Photon Emissions from Cerebral Hemispheres of Fixed Human Brains: Asymmetric Coupling to Geomagnetic Activity and Potentials for Examining Post-Mortem Intrinsic Photon Information

The emissions of biophotons have been considered a ubiquitous property of living systems and their components. We measured the “spontaneous” photon emissions from fixed whole and sectioned human brains within hyper-dark settings. Significant differences in photon counts were measured from different spatial planes. The flux densities were in the order of 2 × 10-12 W per m2. The right hemispheres but not the left hemispheres displayed more photon emissions whose spectral power density profiles exhibited a conspicuous amplitude peak between 7.9 and 8 Hz. Brains measured in the hyperdark (~10-12 W·m2) after removal from the typical lighting of the laboratory emitted more photons than those that had been maintained in the hyperdark for one week. The significant correlation between the numbers of photons emitted from the left hemisphere (but not the right) and global geomagnetic activity also exhibited energy equivalence between the photon flux densities and the geomagnetic shift within the cerebral volumes. These results indicate that what has been assumed to be fixed unresponsive human brain tissue still emits small numbers of photons that may be residuals from ambient light and can potentially interact with global geomagnetic activity. The medical implications for post-mortem intrinsic photonic information based upon the anisotropic microstructures within the hemispheres of the human cerebrum are discussed.

Combined spatial frequency spectroscopy analysis with visible resonance Raman for optical biopsy of human brain metastases of lung cancers

The purpose of this study is to examine optical spatial frequency spectroscopy analysis(SFSA)combined with visible resonance Raman(VRR)spectroscopic method,for thefirst time,to discriminate human brain metastases of lung cancers adenocarcinoma(ADC)and squamous cell carcinoma(SCC)from normal tissues.A total of 31 label-free micrographic images of three type of brain tissues were obtained using a confocal micro-Raman spectroscopic system.VRR spectra of the corresponding samples were synchronously collected using excitation wavelength of 532 nm from the same sites of the tissues.Using SFSA method,the difference in the randomness of spatial frequency structures in the micrograph images was analyzed using Gaussian functionfitting.The standard deviations,calculated from the spatial frequencies of the micrograph images were then analyzed using support vector machine(SVM)classifier.The key VRR biomolecularfingerprints of carotenoids,tryptophan,amide II,lipids and proteins(methylene/methyl groups)were also analyzed using SVM classifier.All three types of brain tissues were identified with high accuracy in the two approaches with high correlation.The results show that SFSA–VRR can potentially be a dual-modal method to provide new criteria for identifying the three types of human brain tissues,which are on-site,real-time and label-free and may improve the accuracy of brain biopsy.

Age-related changes of lateral ventricular width and periventricular white matter in the human brain: a diffusion tensor imaging study

Introduction Aging is the accumulation of multidimensional deterioration of process- ing of biological, psychological, and social changes with expansion over time （Bowen and Atwood, 2004; Grady, 2012）. Aging-related changes are typically accompanied by decline in cognitive function, urinary control, sensory-motor function, and gait ability （Bradley et al., 1991; Bowen and Atwood, 2004; Hedden and Gabrieli, 2004; Grady, 2012; Moran et al., 2012）. In addition, a number of studies have suggested changes in brain structure with normal aging, such as decrease in cortical thickness or increase in ventricular width （Blatter et al., 1995; Tang et al., 1997; Uylings and de Brabander, 2002; Preul et al., 2006; Apostolova et al., 2012）. In particular, ventricular enlargement has been suggested as a structural biomarker for normal aging and progression of some illnesses, such as Alzheimer＇s disease （Blatter et al., 1995; Tang et al.,

Clinical study of recombinant human brain natriuretic peptide in patients with acute myocardial infarction complicating congestive heart failure

Objectives To observe the efficacy and safety of recombinant human brain natriuretic peptide(rh-BNP) on patients with acute myocardial infarction complicating congestive heart failure.Methods 40 patients with acute myocardial infarction complicated by congestive heart failure were randomly divided into control group and treatment group of 20 cases.The control group,15 cases of acute anterior myocardial infarction,5 cases of acute inferior wall myocardial infarction, 15 males and 5 females,aged 55-70 years,mean age 58±12 years;treated 16 cases of acute anterior myocardial infarction,4 cases of acute myocardial infarction,16 males and 4 females,aged 56-70 years,mean age 59±11 years;two groups of age,gender,severity of disease and vascular lesions no significant difference and comparable(P】0.05).Conventional group were given aspirin,clopidogrel, statins,Inotropic,diuretic and vasodilator therapy.In the con- ventional treatment group based on the use of recombinant human brain natriuretic peptide(new bios,Tibet Pharmaceutical Co.,Ltd.Chengdu Nuodikang biopharmaceutical production, usage:1.5μg/Kg intravenous injection(impact), then 0.0075μg-0.01μg/(kg·min)infusion rate).Continuous medication 72 h.The clinical symptoms observed for 3 days in patients before treatment and after treatment,heart rate,blood pressure and left ventricular ejection fraction (LVEF) and tumor necrosis factor(TNF-α),brain natriuretic peptide(BNP) levels were measured.Results In control group,8 cases markedly effect,5 cases effect and 7 cases no effect,the total effective rate was 65%;In treatment group,13 cases markedly effect,6 cases effect and 1 cases no effect,the total effective rate was 95%,compared with two groups P New bios treatment group significantly increased cardiac index(CI) in patients with heart failure and left ventricular ejection fraction(LVEF) than the control group(all P【0.05),further reduce the levels of tumor necrosis (TNF-α) and brain natriuretic peptide(BNP).Conclusions rh-BNP can improve symptoms and heart function,reduced plasma tumor necrosis factor(TNF-α) and BNP levels of acute myocardial infarction patients with congestive heart failure,the treatment safe and reliable.As small sample size observed,larger sample to be accumulated to further evaluate its efficacy and safety.

Orthodontic tooth separation activates the hypothalamic area in the human brain

OBJECTIVES： An animal experiment clarified that insertion of an orthodontic apparatus activated the trigeminal neurons of the medulla oblongata. Orthodontic tooth movement is known to be associated with the sympathetic nervous system and controlled by the nucleus of the hypothalamus. However, the transmission of both has not been demonstrated in humans. The purpose of this study were to examine the activated cerebral areas using brain functional magnetic resonance imaging（MRI）, when orthodontic tooth separators were inserted, and to confirm the possibility of the transmission route from the medulla oblongata to the hypothalamus.METHODS： Two types of alternative orthodontic tooth separators（brass contact gauge and floss） were inserted into the right upper premolars of 10 healthy volunteers. Brain functional T2＊-weighted images and anatomical T1-weighted images were taken.RESULTS： The blood oxygenation level dependent（BOLD） signals following insertion of a brass contact gauge and floss significantly increased in the somatosensory association cortex and hypothalamic area.CONCLUSION： Our findings suggest the possibility of a transmission route from the medulla oblongata to the hypothalamus.

The role of heme-oxygenase-1 in pathogenesis of cerebral malaria in the co-culture model of human brain microvascular endothelial cell and ITG Plasmodium falciparum-infected red blood cells

Objective: To investigate the role of human host heme-oxygenase-1(HO-1) in pathogenesis of cerebral malaria in the in vitro model,Methods: The effect of human host HO-1 [human brain microvascular endothelial cell(HBMEC)] on hemoglobin degradation in the co-culture model of HBMEC and ITG Plasmodium falciparum-infected red cells(i RBC) through measurement of the enzymatic products iron and bilirubin,Results: Following exposure to the HO-1 inducer Co PPIX at all concentrations,the HBMEC cells apoptosis occurred,which could be prominently observed at 15 μM of 3 h exposure,In contrast,there was no significant change in the morphology in the non-exposed i RBC at all concentrations and exposure time,This observation was in agreement with the levels of the enzymatic degradation products iron and bilirubin,of which the highest levels(106.03 and 1 753.54% of baseline level,respectively) were observed at 15 μM vs,20 μM at 3 h vs,24 h exposure,For the effect of the HO-1 inhibitor Zn PPIX,HBMEC cell morphology was mostly unchanged,but significant inhibitory effect on cell apoptosis was seen at 10 μM for the exposure period of 3 h(37.17% of baseline level),The degree of the inhibitory effect as reflected by the level of iron produced was not clearly observed(highest effect at 10 μM and 3 h exposure),Conclusions: Results provide at least in part,insight into the contribution of HO-1 on CM pathogenesis and need to be confirmed in animal model.

Study on threshold segmentation of multi-resolution 3D human brain CT image

In order to effectively improve the pathological diagnosis capability and feature resolution of 3D human brain CT images,a threshold segmentation method of multi-resolution 3D human brain CT image based on edge pixel grayscale feature decomposition is proposed in this paper.In this method,first,original 3D human brain image information is collected,and CT image filtering is performed to the collected information through the gradient value decomposition method,and edge contour features of the 3D human brain CT image are extracted.Then,the threshold segmentation method is adopted to segment the regional pixel feature block of the 3D human brain CT image to segment the image into block vectors with high-resolution feature points,and the 3D human brain CT image is reconstructed with the salient feature point as center.Simulation results show that the method proposed in this paper can provide accuracy up to 100%when the signal-to-noise ratio is 0,and with the increase of signal-to-noise ratio,the accuracy provided by this method is stable at 100%.Comparison results show that the threshold segmentation method of multi-resolution 3D human brain CT image based on edge pixel grayscale feature decomposition is signicantly better than traditional methods in pathological feature estimation accuracy,and it effectively improves the rapid pathological diagnosis and positioning recognition abilities to CT images.

FUNCTIONAL MAPPING OF THE HUMAN BRAIN DURING ACUPUNCTURE WITH MAGNETIC RESONANCE IMAGINGSOMATOSENSORY CORTEX ACTIVATION

Hegu (LI 4) is one of the most frequently used and most important analgesic points in Chinese acupuncture. It is particularly effective for treating disorders of the head and face. According to the meridian theory in Traditional Chinese Medicine (TCM), the Large Intestine Meridian to which it belongs originates in the hand and terminates in the face. This theory is based, however, more on thousands of years of clinical experience rather than on scientific evidence. In our study of acupuncture effects on normal human volunteers with the non-invasive BOLD (blood oxygenation level dependant)technique for FMRI (functional magnetic resonance imaging), we demonstrated widespread effects in the brain during acupuncture at Hegu and Zusanli (ST 36). A finding of special interest was observed in the primary somatosensory cortes (SI) during Hegu acupuncture. In additlon to activation of the area representing the hand in response to the sensory impulses arising from the site of stimulation, activation also occurred in the face representation in all 3 subjects brains studied by coronal brain sections. In one of these subjects activation in the face representation was even stronger than that in the hand representation. Areas representing the neck, trunk and other parts of the upper extremity also exhibited increase in signal intensity, subject to individual variability. As compared with Hegu, such effects were either absent or much weaker with acupuncture at Zusanli (ST 36) or with other forms of sensory stimulation to the hand. Functional mapping of the brain with MRI has provided the first direct evidence in support of the important role of Hegu acupuncture in TCM.

Cerebral organoids exhibit mature neurons and astrocytes and recapitulate electrophysiological activity of the human brain

Multiple protocols have been devised to generate cerebral organoids that recapitulate features of the developing human brain, including the presence of a large, multi-layered, cortical-like neuronal zone. However, the central question is whether these organoids truly present mature, functional neurons and astrocytes, which may qualify the system for in-depth molecular neuroscience studies focused at neuronal and synaptic functions. Here, we demonstrate that cerebral organoids derived under optimal differentiation conditions exhibit mature, fully functional neurons and astrocytes, as validated by immunohistological, gene expression, and electrophysiological, analyses. Neurons in cerebral organoids showed gene expression profiles and electrophysiological properties similar to those reported for fetal human brain. These important findings indicate that cerebral organoids recapitulate the developing human brain and may enhance use of cerebral organoids in modeling human brain development or investigating neural deficits that underlie neurodevelopmental and neuropsychiatric conditions, such as autism or intellectual disorders.

Pain perception and its genesis in the human brain

In the past two decades,pain perception in the human brain has been studied with EEG/MEG brain topography and PET/fMRI neuroimaging techniques.A host of cortical and subcortical loci can be activated by various nociceptive conditions.The activation in pain perception can be induced by physical(electrical,thermal,mechanical),chemical(capsacin,ascoric acid),psychological(anxiety,stress,nocebo) means,and pathological(e.g.migraine,neuropathic) diseases.This article deals mainly on the activation,but not modulation,of human pain in the brain.The brain areas identified are named pain representation,matrix,neuraxis,or signature.The sites are not uniformly isolated across various studies,but largely include a set of cores sites:thalamus and primary somatic area(SI),second somatic area(SII),insular cortex(IC),prefrontal cortex(PFC),cingulate,and parietal cortices.Other areas less reported and considered important in pain perception include brainstem,hippocampus,amygdala and supplementary motor area(SMA).The issues of pain perception basically encompass both the site and the mode of brain function.Although the site issue is delineared to a large degree,the mode issue has been much less explored.From the temporal dynamics,IC can be considered as the initial stage in genesis of pain perception as conscious suffering,the unique aversion in the human brain.

Integrative analyses of RNA editing,alternative splicing,and expression of young genes in human brain transcriptome by deep RNA sequencing

Next-generation RNA sequencing has been successfully used for identification of transcript assembly,evaluation of gene expression levels,and detection of post-transcriptional modifications.Despite these large-scale studies,additional comprehensive RNA-seq data from different subregions of the human brain are required to fully evaluate the evolutionary patterns experienced by the human brain transcriptome.Here,we provide a total of 6.5 billion RNA-seq reads fromdifferent subregions of the human brain.A significant correlation was observed between the levels of alternative splicing and RNA editing,which might be explained by a competition between the molecularmachineries responsible for the splicing and editing of RNA.Younghuman protein-coding genesdemonstrate biased expression to the neocortical and non-neocortical regions during evolution on the lineage leading to humans.Wealso found that a significantly greater number of young human protein-coding genes are expressed in the putamen,a tissue that was also observed to have the highest level of RNA-editing activity.The putamen,which previously received little attention,plays an important role in cognitive ability,and our data suggest a potential contribution of the putamen to human evolution.

Mapping Underlying Maturational Changes in Human Brain

Human brain development is a complex process that continues between birth and maturity, and monitoring the underlying maturational changes at these stages is crucial for our understanding of typical development as well as neurodevelopmental disorders. During the critical periods of brain development, on one hand, many human capacities originate, but on the other hand, a brain undergoing rapid plastic changes may also be vulnerable to neuropsychiatric disorders . Multi-modal magnetic resonance imaging （MRI） has been increasingly used for its ability to noninvasively reveal structural and functional changes in the brain. However, interpretation of the neurobiological processes underlying the findings obtained with MRI is very limited .

Quercetin protects human brain microvascular endothelial cells from fibrillarβ-amyloid_(1–40)-induced toxicity

Amyloid beta-peptides(Aβ) are known to undergo active transport across the blood-brain barrier, and cerebral amyloid angiopathy has been shown to be a prominent feature in the majority of Alzheimer's disease. Quercetin is a natural flavonoid molecule and has been demonstrated to have potent neuroprotective effects, but its protective effect on endothelial cells under Aβ-damaged condition is unclear. In the present study, the protective effects of quercetin on brain microvascular endothelial cells injured by fibrillar Aβ_(1–40)(f Aβ_(1–40)) were observed. The results show that f Aβ_(1–40)-induced cytotoxicity in human brain microvascular endothelial cells(h BMECs) can be relieved by quercetin treatment. Quercetin increases cell viability, reduces the release of lactate dehydrogenase, and relieves nuclear condensation.Quercetin also alleviates intracellular reactive oxygen species generation and increases superoxide dismutase activity. Moreover, it strengthens the barrier integrity through the preservation of the transendothelial electrical resistance value, the relief of aggravated permeability, and the increase of characteristic enzyme levels after being exposed to f Aβ_(1–40). In conclusion, quercetin protects h BMECs from f Aβ_(1–40)-induced toxicity.

De novo identification and quantification of single amino-acid variants in human brain

The detection of single amino-acid variants （SAVs） usually depends on single-nucleotide polymorphisms （SNPs） database. Here, we describe a novel method that discovers SAVs at proteome level independent of SNPs data. Using mass spectrometry-based de novo sequencing algorithm, peptide-candidates are identified and compared with theoretical protein database to generate SAVs under pairing strategy, which is followed by database re-searching to control false discovery rate. in human brain tissues, we can confidently identify known and novel protein variants with diverse origins. Combined with DNA/RNA sequencing, we verify SAVs derived from DNA mutations, RNA alternative splicing, and unknown post-transcriptional mechanisms. Furthermore, quantitative analysis in human brain tissues reveals several tissue-specific differential expressions of SAVs. This approach provides a novel access to high-throughput detection of protein variants, which may offer the potential for clinical biomarker discovery and mechanistic research.

DNA extraction from fresh-frozen and formalin-fixed, paraffinembedded human brain tissue

Both fresh-frozen and formalin-fixed,paraffinembedded（FFPE）human brain tissues are invaluable resources for molecular genetic studies of central nervous system diseases,especially neurodegenerative disorders.To identify the optimal method for DNA extraction from human brain tissue,we compared methods on differently-processed tissues.Fragments of LRRK2 and MAPT（257 bp and 483 bp/245 bp）were amplified for evaluation.We found that for FFPE samples,the success rate of DNA extraction was greater when using a commercial kit than a laboratory-based method（successful DNA extraction from 76%versus 33%of samples）.PCR amplicon size and storage period were key factors influencing the success rate of DNA extraction from FFPE samples.In the fresh-frozen samples,the DNA extraction success rate was 100%using either a commercial kit（QIAamp DNA Micro）or a laboratorybased method（sample boiling in 0.1 mol/L NaOH,followed by proteinase K digestion,and then DNA extraction using Chelex-100）regardless of PCR amplicon length or tissue storage time.Although the present results demonstrate that PCR-amplifiable genomic DNA can be extracted from both fresh-frozen and FFPE samples,fresh brain tissue is recommended for DNA extraction in future neuropathological studies.

β-Estradiol 17-acetate enhances the in vitro vitality of endothelial cells isolated from the brain of patients subjected to neurosurgery

In the current landscape of endothelial cell isolation for building in vitro models of the blood-brain barrier,our work moves towards reproducing the features of the neurovascular unit to achieve glial compliance through an innovative biomimetic coating technology for brain chronic implants.We hypothesized that the autologous origin of human brain mic rovascular endothelial cells(hBMECs)is the first requirement for the suitable coating to prevent the glial inflammato ry response trigge red by foreign neuroprosthetics.Therefo re,this study established a new procedure to preserve the in vitro viability of hBMECs isolated from gray and white matter specimens taken from neurosurge ry patients.Culturing adult hBMECs is generally considered a challenging task due to the difficult survival ex vivo and progressive reduction in proliferation of these cells.The addition of 10 nMβ-estradiol 17-acetate to the hBMEC culture medium was found to be an essential and discriminating factor promoting adhesion and proliferation both after isolation and thawing,suppo rting the well-known protective role played by estrogens on microvessels.In particular,β-estradiol 17-acetate was critical for both freshly isolated and thawed female-derived hBMECs,while it was not necessary for freshly isolated male-derived hBMECs;however,it did countera ct the decay in the viability of the latter after thawing.The tumo r-free hBMECs were thus cultured for up to 2 months and their growth efficiency was assessed befo re and after two periods of cryopreservation.Des pite the thermal stress,the hBMECs remained viable and suitable for re-freezing and storage for several months.This approach increasing in vitro viability of hBMECs opens new perspectives for the use of cryopreserved autologous hBMECs as biomimetic therapeutic tools,offering the potential to avoid additional surgical sampling for each patient.