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Dual-color imaging of nascent blood vessels vascularizing pancreatic cancer in an orthotopic model demonstrates antiangiogenesis efficacy of gemcitabine.

BACKGROUND: The stem cell marker nestin recently has been shown to be expressed in nascent blood vessels in nestin-driven green fluorescent protein (ND-GFP) transgenic nude mice. MATERIALS AND METHODS: In the present study, we visualized by dual-color fluorescence imaging tumor angiogenesis in the ND-GFP transgenic nude mice after orthotopic transplantation of the MIA PaCa-2 human pancreatic cancer line expressing red fluorescent protein. Mice were treated with gemcitabine at 150 mg/kg/dose on days 3, 6, 10, and 13 after tumor implantation. At day 14, mice were sacrificed and mean nascent blood vessel density and tumor volume were calculated and compared to control mice. RESULTS: Nestin was highly expressed in proliferating endothelial cells and nascent blood vessels in the growing tumor. Results of immunohistochemical staining showed that CD31 co-localized in ND-GFP-expressing nascent blood vessels. The density of nascent blood vessels in the tumor was readily quantitated. Gemcitabine significantly decreased the mean nascent blood vessel density in the tumor as well as decreased tumor volume. CONCLUSION: The dual-color model of the ND-GFP nude mouse orthotopically implanted with RFP-expressing pancreatic tumor cells enabled the simultaneous visualization and quantitation of tumor angiogenesis and tumor volume. These results demonstrated for the first time that gemcitabine is an inhibitor of angiogenesis as well as tumor growth in pancreatic cancer. The results have important implications for the clinical application of gemcitabine in this disease.

Angiogenesis Inhibitors↗

Method for modelling cerebral blood vessels and their bifurcations using circular, homogeneous, generalised cylinders.

A method for automatic modelling of blood vessels and their bifurcations from 3D scans of the brain is presented. The method is a three-step procedure. First, a skeleton of the cerebral blood vessels is developed, and then the surfaces of the blood vessels are located using an active contour approach. The active contour approach uses circular homogeneous generalised cylinders (CHGCs) to model the thin, elongated blood vessels. Finally, a novel method for modelling the surfaces of the bifurcations in a vessel tree is presented. The method was tested on simulated data: a computed tomography angiography (CTA) and four magnetic resonance angiography (MRA) volumes. Furthermore, the method was tested on ten magnetic resonance images (MRIs) to demonstrate its robustness. The test on the simulated data indicated that the approach for the surface modelling of vessels had a mean radius error of less than 0.1 mm and a mean localisation error of 0.1 mm. Surface models evaluated by an expert in vascular neurosurgery were found to have a smooth appearance and generally agreed with the image data. The test on the MRI scans indicated that the method performed well in noisy environments.

Brain↗

Decrease of elastic tissue fibres in stem villus blood vessels of the human placenta during IUGR and IUGR with concomitant pre-eclampsia.

In a recent study we described an increase of elastic tissue fibres in blood vessel walls of placental stem villi during pre-eclampsia when compared to uncomplicated pregnancies. Furthermore, the thickness of these blood vessel walls was enhanced in pre-eclampsia. Since it is known that elastic tissue fibres increase in systemic hypertension, it may be assumed that the enhancement of elastic tissue fibres in placental stem villi during pre-eclampsia may be induced by the hypertension. To get further insight into this assumption, we examined the amount of elastic tissue fibres in stem villus blood vessels of placentae of pregnancies complicated by intrauterine growth retardation (isolated IUGR, fourteen cases), a disease without hypertension of the mother and such with pre-eclampsia and concomitant IUGR (IUGR+PE, nine cases). Each study group was compared with uncomplicated pregnancies (twenty-six cases). Unfixed cryostat serial sections were processed for conventional orcein staining and for the demonstration of alpha-actin-immunoreactivity. The intensity of orcein staining of stem villus blood vessel walls was evaluated by a semiquantitative score method. Significant lower intensities of orcein staining were calculated for blood vessel walls of placentae of isolated IUGR (P=0.0007) and IUGR+PE (P=0.0039) when compared to uncomplicated pregnancies each. Additionally, the blood vessel wall thickness of stem villi of isolated IUGR (P=0.0081) and IUGR+PE (P=0.0007) was significantly reduced. In comparison to the above mentioned investigation, our results show that, in contrast to isolated pre-eclampsia, elastic tissue fibres are decreased during pregnancies complicated by IUGR, independently of the occurrence of concomitant pre-eclampsia when compared to uncomplicated pregnancies. From our studies it may be considered that the increase of elastic tissue fibres in placentae of patients with isolated pre-eclampsia may be induced by systemic hypertension. Furthermore, our study underline arguments that IUGR may be an independent disease of the fetus.

Adult↗

Acoustical imaging and processing of blood vessel and the related materials using ultrasound Doppler effect.

In the present paper a method is proposed to measure the degree of the degradation of the elasticity in natural blood vessel and the related materials by using ultrasound Doppler effect. It was found that the deformation rate and its acceleration in the radial direction of the blood vessel can be detected by acoustical imaging and processing using this method. These results were proven to correspond to the degree of the degradation of the elasticity, that is, the degree of viscoelasticity in the blood vessel from the wave versus time pattern detected and its simple analysis. This method was applied to predicting the arteriosclerosis of blood vessels of humans by acoustical imaging and processing uninvadedly, as the characteristics of viscoelasticity in blood vessels.

Algorithms↗

Characterization of sumatriptan-induced contractions in human isolated blood vessels using selective 5-HT(1B) and 5-HT(1D) receptor antagonists and in situ hybridization.

The 5-HT(1B/1D) receptor agonist sumatriptan is effective in aborting acute attacks of migraine and is known to cause constriction of cranial arteries as well as some peripheral blood vessels. The present study set out to investigate whether 5-HT(1B) and/or 5-HT(1D) receptors mediate contractions of the human isolated middle meningeal and temporal arteries (models for anti-migraine efficacy) and coronary artery and saphenous vein (models for side-effect potential). Concentration-response curves were made with sumatriptan (1 nm-100 microm) in blood vessels in the absence or presence of selective antagonists at 5-HT(1B) (SB224289) and 5-HT(1D) (BRL15572) receptors. SB224289 antagonized sumatriptan-induced contractions in all blood vessels, although the antagonism profile was different amongst these blood vessels. In the temporal artery, SB224289 abolished contraction to sumatriptan, whereas in the middle meningeal artery and saphenous vein sumatriptan-induced contractions were blocked in an insurmountable fashion. Moreover, SB224289 acted as a weak surmountable antagonist in the coronary artery (pK(B): 6.4 +/- 0.2). In contrast, BRL15572 had little or no effect on sumatriptan-induced contractions in the four blood vessels investigated. In situ hybridization revealed the expression of 5-HT(1B) receptor mRNA in the smooth muscle as well as endothelial cells of the blood vessels, whereas the mRNA for the 5-HT(1D) receptor was only very weakly expressed. These results show that the 5-HT(1B) receptor is primarily involved in sumatriptan-induced contractions of human cranial as well as peripheral blood vessels.

Adult↗

Pharmacokinetics of an active cadralazine metabolite in plasma and blood vessels of spontaneously hypertensive rats.

The plasma and blood vessel (aorta and mesenteric artery) levels of (+/-)-6-[ethyl(2-hydroxypropyl)amino]-3-hydrazinopyridazine (ISF-2405), an active metabolite of cadralazine, were determined and compared with the changes in blood pressure in spontaneously hypertensive rats after single oral administration of cadralazine. The mean plasma level of ISF-2405 reached a maximum at 0.5 h after oral administration of 3 mg/kg of cadralazine, followed by a rapid elimination with a half-life of 1.0 h, whereas the mean aorta level of ISF-2405 reached a maximum at 4 h after dosing, followed by a slow elimination with a half-life of 6.5 h. The mean mesenteric artery levels of ISF-2405 were almost the same as the mean aorta levels over the time investigated. Both patterns of aorta and mesenteric artery levels of ISF-2405 were in good agreement with those of the hypotensive effects. These findings substantiate the assumption that the slow onset and long duration of the pharmacological effect of cadralazine is closely related to the distribution pattern of the active metabolite ISF-2405 in blood vessels, a target tissue of antihypertensive vasodilator drugs.

Animals↗

Characterization of a vasopressin-like peptide in rat and bovine blood vessels.

The present study examined the possibility that blood vessels of the rat and cow contain arginine vasopressin (AVP) and that the vascular stores of this potent vasoconstrictor are of local, rather than blood-borne, origin. Vessels from rat or cow were homogenized in acid, and the supernatants were assayed for AVP by radioimmunoassay. AVP immunoreactivity was detected in all vessels taken from the rat including aorta, mesenteric artery, renal artery, vena cava, and renal vein, as well as atria of the heart and also in cow aorta. Blood vessels from hypophysectomized and Brattleboro rats also contained AVP at levels similar to those of intact control rats. Further characterization of this immunoreactive material showed that, in both radioimmunoassay and radioreceptor assay, vascular extracts produced competition curves, which were parallel to those of synthetic AVP. Additionally, immunoreactive AVP in aortic extracts comigrated with synthetic AVP and pituitary extract on both high-pressure liquid chromatography and Sephadex G-25 chromatography. Furthermore, intravenous administration of the aortic extract produced pressor responses in conscious rats, and these responses were significantly attenuated by pretreatment with a specific AVP V1-receptor antagonist. These data demonstrate that blood vessels contain an AVP-like peptide that appears indistinguishable from authentic AVP and further suggest that this vascular peptide is of local origin.

Animals↗

Presence of angiotensin converting enzyme in the adventitia of large blood vessels.

BACKGROUND: Angiotensin converting enzyme (ACE) is present in the endothelial cells of all vascular beds. There are, however, many reports of converting enzyme activity in blood vessels not associated with the endothelium. METHODS: ACE was localized in large blood vessels of a number of mammals by in vitro autoradiography using the radioligand 125I-351A. To characterize this binding further, immunohistochemistry was performed on rabbit aorta using polyclonal antisera raised to two different preparations of rabbit lung ACE. RESULTS: In all of the blood vessels studied, which included the rabbit pulmonary artery, rabbit, dog and sheep aorta, human internal mammary artery and human saphenous vein, high levels of radioligand binding were found in endothelial cells, as expected. In addition, a very high density of punctate binding was observed interspersed between diffuse moderate labelling in the adventitia. Immunoreactivity was confined to the endothelium of both the intima and the vasa vasorum of the adventitia. The immunostaining correlated well with the autoradiography. The ACE inhibitors lisinopril and perindoprilat displayed similar high affinities in competing for the binding of 125I-351A to the endothelium and adventitia of the sheep aorta, suggesting that at these two sites the radioligand was binding to ACE. CONCLUSIONS: We find that ACE in the adventitia of large blood vessels is confined to the vaso vasorum. The results of this study help to explain the findings of many studies that ACE activity persists in endothelium-denuded blood vessels and also reveals a source of ACE distant from the luminal endothelial surface.

Angiotensin-Converting Enzyme Inhibitors↗

[Development of artificial blood vessel suitable for cerebrovascular surgery: improvement in the mechanical properties].

Saphenous vein interposition grafts have been commonly used for the reconstruction of occlusive lesions in the extracranial cerebral arteries, such as carotid or vertebral arteries. In contrast, cerebral revascularization using an artificial blood vessel has not been so common. This is due to the fact that conventional artificial blood vessels have been too stiff for use in the cerebrovascular surgery. Another reason is that the patency rate of small-caliber artificial blood vessels has usually been inferior to that found in autologous vein grafts. The purpose of this study was to develop a pliable and compliant artificial blood vessel suitable for the reconstructive cerebrovascular surgery. This new artificial blood vessel is made of polyurethane, porous in structure (porous polyurethane). Thus, multiple small-sized pores exist both in the inner and outer surfaces, and in the wall of the graft. To test its mechanical properties, stress-strain curves and compliance were evaluated. In comparison to expanded polytetrafluoroethylene graft, which has been one of the most commonly used artificial blood vessels in the cardiovascular surgery, the mechanical properties of the porous polyurethane graft more closely resembled those of the common carotid artery in dogs. This means not only that it can be maneuvered with technical ease for anastomosis but also that there is a reduction of compliance mismatch between the host vessel and the artificial vessel. Compliance mismatch has been documented as a major factor in the inducement of intimal hyperplasia, which causes a delayed occlusion of the grafts. In the in vivo animal experiments, a porous polyurethane graft (2.3-3.2 mm in diameter, 40-50 mm in length) was transplanted into the common carotid artery of dogs. The mean size of pore varied from 0 (group 1), 1.7 (group 2), 4.4 (group 3), 5.5 (group 4), 7.4 (group 5), and 30.0 microns (group 6). In the follow-up study conducted 2 months after the transplantation, all grafts in group 4, 5, and 6 were shown to be patent, whereas no patency could be obtained in groups 1, 2 and 3. In addition, all grafts in group 6 were patent 6 months after transplantation, though one showed remarkable stenosis due to anastomotic intimal hyperplasia. Histological examination showed that the luminal surface of the graft was covered with monolayer of endothelium-like cells. Neoadventitia made of collagen developed around the external surface of the graft. Foreign body reactions were minimal. In conclusion, a porous polyurethane graft could be developed, which has improved mechanical properties suitable for the reconstructive surgery of the extracranial cerebral arteries.

Animals↗

Evidence for production and functional activity of nitric oxide in seminiferous tubules and blood vessels of the human testis.

Previous studies have demonstrated that nitric oxide (NO) influences Leydig cell function. Here we provide evidence for NO production and activity in seminiferous tubules and blood vessels of the human testis. By immunohistochemistry, the soluble guanylyl cyclase (sGC), the intracellular NO receptor, and the second messenger, cyclic guanosine monophosphate (cGMP), were detected in myofibroblasts of the peritubular lamina propria in Sertoli cells, as well as in endothelial and smooth muscle cells of testicular blood vessels. Performed with isolated tubules and blood vessels, the biological activity of sGC could be proved by cGMP generation in response to treatments with the NO donor, sodium nitroprusside. The endothelial and neuronal subtypes of NO synthase (NOS) were localized immunohistochemically to the same cell types that express sGC and cGMP. In isolated tubules and vessels, the presence of endothelial NOS and neuronal NOS was confirmed by immunoblotting, and NOS activity was demonstrated by decreased cGMP production upon incubation with the NOS inhibitor L-nitro arginine methylester. These findings show that peritubular cells, Sertoli cells, and testicular blood vessels may be sites of NO production and activity, possibly involved in relaxation of seminiferous tubules and blood vessels to modulate sperm transport and testicular blood flow, respectively.

Adult↗

Relaxant effects of butylidenephthalide in isolated dog blood vessels.

We investigated the reason why butylidenephthalide (Bdph) can have an antianginal effect without changing blood pressure in conscious rats. Isolated dog coronary artery (CA), femoral vein (FV), femoral artery (FA), and mesenteric artery (MA) were used to evaluate the relaxant effects of Bdph. Bdph concentration-dependently relaxed isolated CA, FV, FA, and MA precontracted by KCl (60 mM) and phenylephrine (phe, 5 microM) with the exception that CA was precontracted by prostaglandin F 2 alpha (PGF 2 alpha, 2 microM) instead of phe. The potency order of Bdph to these blood vessels was FV > CA > FA > or = MA. Bdph also concentration-dependently and non-competitively inhibited cumulative KCl (5 - 120 mM)- and phe (0.1 - 100 microM)-induced contractions in normal, and inhibited cumulative Ca 2+-induced contractions in depolarized blood vessels. The potency order of Bdph to these blood vessels was FV congruent with CA > FA congruent with MA. Bdph (0.02 - 0.04 mM) concentration-dependently and leftward-shifted the log concentration-response curves in parallel and significantly increased the pD 2 value of forskolin, but not nitroprusside in FV. Bdph (0.1 mM) did both in CA. Bdph (0.225 - 0.45 mM) did the opposite to that of nitroprusside, but not forskolin, in FA. Bdph (0.45 - 0.9 mM) did neither in MA. Bdph (0.1 - 1 mM) significantly inhibited cAMP- but not cGMP-PDE activities in these four blood vessels, suggesting that Bdph more selectively inhibited the former in these tissues. The above results suggest that the higher potencies of Bdph on FV and CA than on FA and MA, may be interpreted as the reason why Bdph is useful in the treatment of angina pectoris without changing blood pressure, after Bdph administration in vivo, because the venoreturn may be reduced and the coronary flow may be increased without affecting the arterioles, such as MA, the main determinant of blood pressure. Abbreviations. Bdph:butylidenephthalide Phe:phenylephrine PGF 2alpha :prostaglandin F 2alpha CA:coronary artery FV:femoral vein FA:femoral artery MA:mesenteric artery cAMP:adenosine 3',5'-cyclic monophosphate cGMP:guanosine 3',5'-cyclic monophosphate PDE:phosphodiesterase

Angina Pectoris↗

The blood vessel wall as an insulin target tissue.

Study of the effects of insulin and insulin-like growth factors on blood vessel walls has been expedited by the ability to isolate and grow components of large and small blood vessels in pure culture. This paper summarizes current knowledge about the effects of insulin and related molecules on blood vessel derived cells, with an emphasis on: (1) peptide receptors, their regulation, and transport and processing of molecules through the endothelium; (2) metabolic and (3) mitogenic effects of insulin and related peptides on the blood vessel wall, with a discussion of major differences between large and small vessel endothelia; and (4) interactions of insulin with barrier endothelia, for example cerebral microvascular endothelium, site of the blood-brain barrier.

Aminoisobutyric Acids↗

[Change of NOS activity in hypoxia and cold-induced blood vessels damage and its biological significance].

OBJECTIVES: To explore the therapeutic methods for hypoxia and cold induced blood vessel damage by observing the changes of NO synthase (NOS) activity in hypoxia and development of coldm induced blood vessels damage, and studying the effects of SOD and vitamin C on the changes of NOS activity. METHODS: The main artery was isolated from Wistar rats and exposed to hypoxia and/or cold environment in vitro. LDH activity was measured using an automatic biochemical analyzer and NOS and SOD activities in blood vessels were analyzed respectively by Griess assay and adrenaline autoxidantion method. RESULTS: NOS activity in hypoxia, cold and hypoxia-cold damaged vessels decreased by 18.2%, 19.1% and 25.9% respectively after exposure as compared with the control vessels. The changes of NOS activity were time dependent. The NOS activity in damaged vessels was negatively correlated with LDH activity in the culture medium of the respective vessels. The SOD activity in hypoxia and/or cold damaged vessels also decreased significantly. Giving SOD (200 u/ml) or vitamin C (50 mg/ml) immediately after hypoxia and/or cold exposure increased the NOS activity and SOD activity in the damaged vessels significantly, as compared with the non-treatment controls. CONCLUSIONS: The decrease of NOS activity is closely related to the development of damage induced by hypoxia and/or cold injury. SOD and vitamin C could protect the blood vessels exposed to hypoxia and/or cold from damage, possibly by increasing NOS activity.

Animals↗

The relationship of glands to thick-wall blood vessels as a marker of invasion in endocervical adenocarcinoma.

The recognition of early invasion in endocervical adenocarcinomas can be difficult. We evaluated the proximity of endocervical glands and their pattern of infiltration in relation to thick-wall blood vessels as potential markers of invasion in a series of invasive endocervical adenocarcinomas removed by cone biopsy or hysterectomy. Routinely stained slides were examined from 50 invasive endocervical adenocarcinomas (37 of usual type and 13 of minimal deviation type), 26 noninvasive lesions (14 cases of adenocarcinoma in situ, 7 cases of hyperplasia, 4 cases of tunnel clusters, 1 adenomyoma), and 20 normal cervices, including 7 with deep nabothian cysts. The range of vessel wall thickness using an ocular micrometer was recorded in each specimen. A blood vessel with a wall > or =36-microm thick was defined as thick-walled. Both the blood vessel wall thickness and distance to the closest gland was recorded for each case. The median distance from a thick-wall blood vessel to a gland in the invasive tumors was 30 microm compared with 168 microm in the noninvasive group. In both groups, the measured blood vessel wall had a median thickness of 42 microm. The pattern of infiltration of glands around the thick-wall vessels was classified as "circumferential," in which multiple glands circumferentially enveloped the vessel, or "molded," in which single or multiple distorted glands were molded around the vessel. The circumferential and molded patterns were identified in 35 (70%) and 10 (20%) of 50 invasive adenocarcinomas, respectively. Of the 26 lesions in the noninvasive group, only 3 (6%) had a circumferential pattern and none had a molded pattern. Importantly, none of the cases in the noninvasive group showed either pattern in the presence of glandular atypia. In conclusion, close proximity of glands to thick-wall blood vessels (distance from the closest gland to a thick-wall vessel less than or equal to the thickness of the vessel wall) seems to be a useful feature in the diagnosis of invasive endocervical adenocarcinoma. This feature, in combination with certain glandular growth patterns and cytologic atypia, can be highly suggestive, if not diagnostic, of invasive carcinoma.

Adenocarcinoma↗

Transillumination optical tomography of tissue-engineered blood vessels: a Monte Carlo simulation.

A Monte Carlo technique has been developed to simulate the transillumination laser computed tomography of tissue-engineered blood vessels. The blood vessel was modeled as a single cylinder layer mounted on a tubular mandrel. Sequences of images were acquired while rotating the mandrel. The tomographic image was reconstructed by applying a standard Radon transform. Angular discrimination was applied to simulate a spatial filter, which was used to reject multiply scattered photons. The simulation results indicated that the scattering effect can be overcome with angular discrimination because of the thin tissue thickness. However, any refractive-index mismatch among the tissue, the surrounding media, and the mandrel could produce significant distortions in the reconstructed image.

Animals↗

Measurement of retinal blood vessel width using computerized image analysis.

We report a new method for quantitating retinal blood vessel width from fluorescein angiogram negatives using a computerized image analyser. Interuser and intrauser variability were 3 to 4 times lower than reported with older methods. In addition, model blood vessels were used to test the accuracy of our method under four clinically relevant conditions: variations in vessel width, fluorescein concentration, flash intensity and background fluorescence. Although computed measurements were affected by vessel width, fluorescein concentration and flash intensity, the linearity of the actual versus computed width was maintained during wide variations in all four conditions (r = 0.95, P less than 0.0001). Accuracy was best achieved at a flash intensity of 150 W/sec and a fluorescein concentration of 45 micrograms/ml. The results of this study provide a better understanding of factors affecting the apparent width of retinal blood vessels in fluorescein angiograms. This technique should be useful in rapidly obtaining accurate measurements of blood vessel width from fluorescein angiograms.

Animals↗

[Repair of defects at both ends of blood vessels in extremities with a great disparity in diameter by vein transplantation].

OBJECTIVE: To repair defects at both ends of the blood vessels with a considerable disparity in the diameter of the both sides or with a large diameter in extremities by phleboplasty of branched and double autogenous veins. METHODS: Three kinds of phleboplasty: funnel-shaped, raincape-shaped and transposed Y-shaped were designed. Experiments in fresh blood vessels in vitro were completed successfully. These methods were used clinically to repair injured external iliac veins, femoral arteries and veins, and popliteal arteries and veins, to replant severed fingers and to transplant toenail flaps on thumbs by harvesting autogenous great saphenous veins, small saphenous veins and forearm veins in 36 cases, including 35 cases in emergency operation and 1 case in selective operation. The length of The phleboplasty of funnel-shaped could enlarge the grafted blood vessels ranged from 1.0 cm to 15.0 cm. RESULTS: The phleboplasty of funnel-shaped could enlarge the diameter by 1.0-1.25 times in anastomotic stomas. The phleboplasty of raincape-shaped could enlarge the diameter large enough to meet the demands for various blood vessels in extremities. The phleboplasty of transposed Y-shaped could provide large vein transplants. In 36 grafted veins, 35 were in patency. The blood supply in extremities was normal. CONCLUSION: The funnel-shaped and raincape-shaped phleboplasty of branched veins can enlarge the anastomotic stomas of grafted veins. The transposed Y-shaped phleboplasty of double femoral veins is an ideal way to repair injured primary blood vessels with a considerable disparity in the diameter of the both sides or with a large diameter in extremities.

Adolescent↗

[Clinicopathological study of the fundic varices--blood vessel structure in gastric paries].

The structures of blood vessels in 18 cases with fundic varices (FV) were studied by injecting gelatin-added barium into varicose vessels of resected specimens. There was no direct connection outside the gastric paries between the inflow path and the outflow path of FV. Just outside the gastric paries, the outflow path was not branched. Blood vessels which formed FV were found in the submucous layer, but not in the lamina propria mucosae. FVs were classified into two types according to the ramification form of FV-forming vessels inside the gastric paries: Type I, with only a few rami observed, and type II, with many rami recognized. In this study, type I FV accounted for about 78%, and type II FV about 22% of 18 cases observed. These findings deserve full consideration in the diagnosis and treatment of FV.

Adult↗