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Biomedical subjects

Y L Chen

Publications and source records attributed to Y L Chen.

At least 73 records · Page 4Linked to original sources

[Modulation of nerve growth factor on wound healing of burn].

OBJECTIVE: To investigate the effect of nerve growth factor(NGF) on the burn wound healing and to study the mechanism of burn wound healing. METHODS: Six domestic pigs weighting around 20 kg were used as experimental animals. Twenty-four burn wound, each 2.5 cm in diameter, were induced on every pigs by scalding. Three different concentrations of NGF, 1 microgram/ml, 2.5 micrograms/ml, 5 micrograms/ml were topically applied after thermal injury, and saline solution used as control group. Biopsy specimens were taken at 3, 5 and 9 days following treatment and immunohistochemistry method was used to detect the epidermal growth factor(EGF), EGF receptor (EGF-R), NGF, NGF receptor (NGF-R), NGF, NGF-R, CD68 and CD3. RESULTS: The expression of EGF, EGF-R, NGF, NGF-R CD68 and CD3 were observed in the experimental group, especially at 5 and 9 days, no expression of those six items in the control group. CONCLUSION: NGF can not only act directly on burn wound, but also modulate other growth factors on the burn wound to accelerate the healing of burn wound.

Animals↗

[Isolation of a gene related to trichosanthin-induced apoptosis (GRETA)].

Disturbance of the apoptosis-related signaling pathway is regarded as one of the critical factors for tumorigenesis. Isolation of the genes involved in the process of apoptosis would be thereby helpful to explore the mechanism of tumor transformation and to develop novel therapeutic approaches. Here we report a gene fragment GRETA, the gene related to trichosanthin-induced apoptosis, isolated from a leukemia cell line U937 undergoing apoptosis induced by a plant protein Trichosanhin (TCS). A 293bp segment of GRETA was revealed to be 78.3% homologous to Bruton's tyrosine kinase at nucleic acid level. And Northern blot analysis showed that three messengers of RNA with the size of about 0.8-kb, 2.0-kb and 7.0-kb, respectively, were detected in TCS-untreated U937 cells when probed with GRETA, but there were only 0.8-kb and 2.0-kb transcripts appeared in apoptotic U937 cells. In addition, the abundance of each transcript changed apparently. The 0.8-kb transcript, for example, was the main band in Northern analysis in apoptotic U937 cells while was only detected marginally in TCS-untreated cells. These data suggested a possible relationship between the alternative splicing patterns of GRETA and the apoptosis.

Apoptosis↗

Red wine inhibits monocyte chemotactic protein-1 expression and modestly reduces neointimal hyperplasia after balloon injury in cholesterol-Fed rabbits.

BACKGROUND: Wine consumption decreases the risk of myocardial infarction. Intimal hyperplasia contributes to restenosis after angioplasty. Local ethanol delivery inhibits intimal hyperplasia after balloon injury in rabbit iliac and pig coronary arteries. The effects of wine consumption on intimal response and monocyte chemotactic protein-1 (MCP-1) expression were studied in cholesterol-fed rabbits. METHODS AND RESULTS: Male rabbits were fed a 2% cholesterol diet together with red wine (12.5% vol, 5 mL/kg body wt per day; n=7), white wine (13.3% vol, 5 mL/kg body wt per day; n=7), or no wine as a control (n=8) for 6 weeks. A balloon injury of the abdominal aorta was performed at the end of the third week. Abdominal aortas were harvested at the end of 6 weeks. Neointimal hyperplasia was measured morphometrically. MCP-1 expression was determined by Northern blot, in situ hybridization, and immunohistochemistry. Rabbits fed red wine had significantly less neointimal hyperplasia than did control rabbits (intima/media area ratio 0.59+/-0.05 [red wine group] versus 0.79+/-0.07 [control group], P<0.05). However, rabbits fed white wine showed a trend (but not significant) toward less intimal response compared with control rabbits (intima/media area ratio 0.65+/-0.04 [white wine group] versus 0.79+/-0.07 [control group], P=0.165). Both red wine and white wine significantly reduced MCP-1 mRNA and protein expression in the aorta. CONCLUSIONS: Long-term consumption of red wine and white wine inhibits MCP-1 expression, and in the small number of animals studied, red wine modestly reduces neointimal hyperplasia. Since red wine exhibits higher antioxidant capacity than does white wine, the decreased intimal response might be partly attributed to its antioxidant effects.

Animals↗

Reconstitution of acid-denatured holomyoglobin studied by time-resolved electrospray ionization mass spectrometry.

Time-resolved electrospray ionization (ESI) mass spectrometry (MS) is a new technique for studying the kinetics of protein folding reactions. It can monitor both changes in the protein conformation and the loss or binding of protein ligands as a function of time. Time-resolved ESI MS was previously used to monitor the acid-induced unfolding of holomyoglobin (hMb). The native form of this protein is characterized by a tightly folded conformation and a heme group that is noncovalently attached to the protein. Acid-induced denaturation induces substantial unfolding of the polypeptide chain and disruption of the heme-protein interactions. In this work, time-resolved ESI MS is used to study the reverse reaction, i.e., reconstitution of acid-denatured hMb. To examine the mechanism and the kinetics of this reaction, a continuous-flow setup with two sequential mixing steps was developed. The data presented in this work show that reconstitution involves the formation of various short-lived intermediates such as tightly folded myoglobin without a heme group and several nativelike forms of the protein that are bound to more than one heme. The occurrence of these transient states is most likely due to the rapid aggregation of free heme in solution.

Acetic Acid↗

Vertebral centroid measurement of lumbar lordosis compared with the Cobb technique.

STUDY DESIGN: An observational study in which vertebral centroid measurement of lumbar lordosis, developed in this study, was used to examine lumbar curvature. The intra- and interobserver reliability of the vertebral centroid measurement of lumbar lordosis and the Cobb technique were compared. OBJECTIVES: To evaluate the reliability of a new method of measuring lumbar lordosis and to examine the changes in the lordotic curve from 0 degree to 90 degrees flexion of the trunk. SUMMARY OF BACKGROUND DATA: Several different methods are used to measure lumbar lordosis. The Cobb technique, based on measurement of vertebral endplates, is the method most frequently adopted for clinical diagnosis. However, because of the variations in the vertebral endplate architecture, the vertebral surface angle is difficult to identify. This reduces the reliability of the Cobb technique. METHODS: Lateral radiographs of 16 study participants were taken from the upright position to a trunk flexion of 90 degrees in 30 degrees increments. The lumbar lordotic curve was measured by three observers individually using two applications of the traditional Cobb technique and the vertebral centroid measurement of lumbar lordosis. RESULTS: Correlation coefficients of lumbar lordosis between the two methods ranged from 0.589 to 0.772 with participants standing upright (all P < 0.05). Interobserver reliability coefficients were 0.903 for vertebral centroid measurement of lumbar lordosis, 0.826 for Cobb (L1-L5), and 0.784 for Cobb (L1-S1), although the three measurements all revealed an excellent intraobserver reproducibility (r greater than 0.9). The vertebral centroid measurement of lumbar lordosis showed the smallest mean absolute differences between any two observers' measurements (< 1.7 degrees). CONCLUSIONS: The findings from this study indicate that the vertebral centroid measurement of lumbar lordosis is more reliable than the Cobb method for assessing lumbar lordosis. The vertebral centroid measurement of lumbar lordosis also can be used to evaluate the actual lumbar curvature in outline at various angles of trunk flexion.

Adult↗

Oxytocin and vasopressin constrict rat isolated uterine resistance arteries by activating vasopressin V1A receptors.

Both oxytocin and vasopressin cause potent and long-lasting vasoconstriction of uterine arteries from several species, including humans, and the resulting tissue ischemia is thought to be involved in the pathogenesis of primary dysmenorrhea. We have studied the effects of oxytocin and vasopressin in isolated resistance arteries (diameter, 90-120 microm) from non-pregnant rat uteri using two potent and selective receptor antagonists, SR 49059, a selective vasopressin V1A antagonist, and atosiban, a selective oxytocin antagonist. Uterine arteries with intact endothelium were mounted in a microvessel chamber, and pressurized to 75 mm Hg to allow the development of myogenic tone. Both vasopressin and oxytocin elicited a concentration-dependent vasoconstriction with a similar maximum effect (i.e., total vessel occlusion). The EC50 was 0.44 +/- 0.02 and 25 +/- 3.1 nM for vasopressin and oxytocin, respectively. Thus, vasopressin was 57-fold more potent than oxytocin. Schild analysis indicated that SR 49059 yielded a similar pA2 value against vasopressin-induced (pA2 = 8.96 +/- 0.60) or oxytocin-induced (pA2 = 9.06 +/- 0.23) contractions, suggesting that both agonists activated the vasopressin V1A receptor. In addition, atosiban (10(-7) M), a selective antagonist of the oxytocin receptor in the rat, did not antagonize the effect of vasopressin and oxytocin, showing that the oxytocin receptor is not involved in the response. In conclusion, these results suggest that V1A receptor stimulation is responsible for the vasoconstricting effects of both vasopressin and oxytocin in small diameter resistance arteries from the rat uterus.

Animals↗

Geometric measurements of the lumbar spine in Chinese men during trunk flexion.

STUDY DESIGN: An analysis of the geometric data of the lumbar spine in Chinese men. Lateral radiographs were obtained of 16 men in the upright position to a trunk flexion of 90 degrees in 30 degrees increments. OBJECTIVES: To establish reference data concerning the geometry of the lumbar spine for various degrees of trunk flexion. SUMMARY OF BACKGROUND DATA: In previous studies, the morphometry and alignment of the lumbar spine have been investigated. Reports of systematic analyses of the lumbar spine in trunk flexion have lacked adequate radiographic data. Moreover, few studies have focused on measuring lumbar spine geometric data in Chinese individuals. METHODS: A total of 67 landmarks on each radiograph were identified. They were marked by investigators and digitized using a HyperSpace digitizing system (V.17: Mira Imaging Inc., Salt Lake City, UT). The geometric configuration of the vertebrae and discs then was derived from the digitized points. RESULTS: The dimensions of the vertebrae and disc on the lumbar spine obtained in this study were similar to those of previous studies. The motion of the lumbosacral spine had the greatest contribution in trunk flexion, approximately 90% (i.e., rotating at the rate of 9 degrees for each 10 degrees of trunk flexion) when the trunk was flexed from 30 degrees to 60 degrees. After that, the hips were dominant in accomplishing the trunk flexion from 60 degrees to horizontal. This motion patterns may be useful for making clinical diagnoses of lumbar function in Chinese men. CONCLUSIONS: No obvious interracial differences were found in the geometric data found in this study, which suggests that morphometric data obtained from Caucasian individuals may be applied to Chinese patients for clinical purposes.

Adult↗

Simian virus 40 T antigen induces p53-independent apoptosis but does not suppress erbB2/neu gene expression in immortalized human epithelial cells.

Previously, we have shown that simian virus 40 (SV40) T antigen can directly cause apoptosis in immortalized human epithelial cells under normal growth conditions. In this study, we further characterized the mechanism of T-antigen-mediated apoptosis involving p53 and whether T antigen can suppress erbB2/neu gene expression. Our results show the differential regulation of erbB2/neu gene expression in different cell clones in response to T antigen transgene, indicating that the regression of erbB2/neu gene by SV40 T is cell-type dependent. Our previous study reported T-antigen-induced apoptosis in p53 mutant cells; however, in this study we find increased levels of p53 protein in T-antigen-containing cells. Therefore, we examined the transactivation function of p53 in these cells. Our data show the failure to transactivate p53, suggesting that increased p53 protein in T antigen expressing cells is functionless at least for transcriptional activation.

Antigens, Viral, Tumor↗

Cyclic strain stimulates monocyte chemotactic protein-1 mRNA expression in smooth muscle cells.

Hemodynamic forces are important determinants for the formation of atherosclerotic plaques. The recruitment of circulating monocytes into the arterial wall is an important step during atherogenesis. Monocyte chemotactic protein-1 (MCP-1) has been shown to be a key factor for monocyte transmigration. This study examined the effects of cyclic strain on MCP-1 mRNA expression levels of cultured rat aortic smooth muscle cells. The MCP-1 mRNA levels of aortic smooth muscle cells first increased as the duration of cyclic strain increased, reaching the maximum at 6-12 h, maintained at high levels throughout the 48-h strain period. To explore signaling pathways mediating cyclic strain-stimulated MCP-1 mRNA expression, we examined the involvement of tyrosine kinase and protein kinase C (PKC). Tyrosine kinase inhibitors, genistein and tyrphostin 51, at 50 microM blocked cyclic strain-stimulated MCP-1 mRNA expression. Preincubation with a PKC activator, phorbol 12-myristate 13-acetate (PMA), 2 microM, for 24 h to downregulate PKC did not decrease cyclic strain-induced MCP-1 mRNA expression. A 6-h incubation with 0. 1 microM PMA to activate PKC, which stimulated MCP-1 expression when applied alone, abolished the stimulatory effects of cyclic strain. A specific PKC inhibitor, calphostin C (0.1 microM), diminished cyclic strain-stimulated MCP-1 mRNA expression. Angiotensin II at 10 or 1,000 nM induced a moderate upregulation of MCP-1 mRNA, and no synergistic effects were observed between angiotensin II and cyclic strain. These results indicate that cyclic strain stimulates MCP-1 mRNA expression in smooth muscle cells through signaling pathway(s) mediated by tyrosine kinase activation.

Angiotensin II↗

Electrophoretic characterization of a novel cysteine protease produced by Vibrio harveyi.

Electrophoretic characterization of a novel cysteine protease produced by pathogenic luminous Vibrio harveyi, originally isolated from diseased tiger prawn Penaeus monodon in Taiwan, is demonstrated in the present study using native polyacrylamide gel electrophoresis (native PAGE), sodium dodecyl sulfate-PAGE (SDS-PAGE), crossed immunoelectrophoresis (CIE) and isoelectric focusing (IEF) gels. The protease has a pI of 6.4 and exhibits a fast-migrating feature in native-PAGE and CIE gels indicating that it is a negatively charged protease. The protease electrophoresed as a 22 kDa protein band in native- and SDS-PAGE (in SDS - buffer with or without the presence of 2-mercaptoethanol) while it electrophoresed as a 38 kDa protein band in SDS-PAGE when the samples were boiled for 10 min prior to electrophoresis. The results reveal that the enzyme is an SDS-resistant monomeric protease and its high negative charge is not influenced by SDS (detergent) without boiling the sample. The present results are useful in determining proteins of similar nature to this unique cysteine protease.

Cysteine Endopeptidases↗

Monocyte chemotactic protein-1 gene and protein expression in atherogenesis of hypercholesterolemic rabbits.

Monocyte adherence to the endothelium and subsequent migration into the subendothelial space are important early events in atherogenesis. Monocyte chemotactic protein-1 (MCP-1) has been shown to be highly expressed in both human atheroma and advanced atherosclerotic lesions of experimental animals. To establish the temporal correlation between MCP-1 expression and plaque development, we examined the expression of MCP-1 during atherogenesis of hypercholesterolemic rabbits using Northern blot analysis, in situ hybridization, and immunohistochemistry. New Zealand White rabbits were fed with 2% cholesterol-containing diet for 1 day, 3 days, 1 week, 3 weeks or 6 weeks. The plasma levels of total cholesterol were significantly increased 3 days after cholesterol feeding and continued to increase during the entire cholesterol-feeding period. Northern blot analysis showed that MCP-1 mRNA levels remained unchanged following cholesterol feeding for up to 1 week, were higher than control levels at 3-week and increased even higher at 6-week. In situ hybridization showed that after 3 weeks of cholesterol feeding, MCP-1 mRNA expression was up-regulated in newly-formed fatty streaks and parts of tunica media in the presence or absence of fatty streaks. At 6-week, pronounced MCP-1 mRNA expression was detected with similar distribution. In contrast, MCP-1 mRNA was detected only in a few endothelial cells and adventitia in control and experimental groups feeding cholesterol up to 1-week. Immunostaining of serial sections indicated that MCP-1 was expressed by macrophages and smooth muscle cells in rabbits fed with cholesterol for 3 or 6 weeks. No MCP-1 was detected in intima or media in all other groups. These results show that a lag period exists between serum cholesterol increase and upregulation of MCP-1 expression, suggesting that cholesterol modifications (e.g. oxidation) are required to stimulate MCP-1 expression. In addition, MCP-1 expressed by both macrophages and smooth muscle cells during the initial stages of atherosclerosis is likely to contribute to the development of fatty streaks in hypercholesterolemic rabbits.

Animals↗

An unusual arrangement of pur and lpx genes in the photosynthetic purple sulfur bacterium Allochromatium vinosum.

The nucleotide sequence of a 1634 bp DNA fragment from the photosynthetic purple sulfur bacterium Allochromatium vinosum contains one complete and two partial open reading frames. Sequence comparisons to genes from other organisms suggest that this A. vinosum DNA fragment contains, starting from the 5' end, the following: (1) 234 bp at the 3' end of the A. vinosum purH gene, coding for 78 amino acids at the C-terminus of the bi-functional 5'-phosphoribosyl-5-aminoimidazole-4-carboxamide formyltransferase/IMP cyclohydrolase (EC 2.1.2.3), an enzyme involved in de novo purine biosynthesis; (2) 777 bp of the A. vinosum lpxA gene, coding for all 259 amino acids of the UDP-N-acetylglucosamine-O-acyltransferase, an enzyme involved in lipid A biosynthesis; and (3) 567 bp at the 5' end of the A. vinosum purD gene, coding for 189 amino acids at the N-terminus of 5'-phosphoribosyl glycinamide synthetase (EC 6.3.4.13), a second enzyme involved in de novo purine biosynthesis. The presence of a gene coding for an enzyme involved in lipid A biosynthesis between two genes coding for enzymes of the de novo purine biosynthesis pathway represents a unique arrangement of these genes.

Acyltransferases↗

Enterotoxigenic profiles and polymerase chain reaction detection of Bacillus cereus group cells and B. cereus strains from foods and food-borne outbreaks.

Bacillus cereus is one of the important food pathogens. Since B. cereus group cells, such as B. cereus, B. thuringiensis, B. anthracis and B. mycoides, share many phenotypical properties and a high level of chromosomal sequence similarity, it is interesting to investigate the virulence profiles for B. cereus group cells, including B. cereus strains isolated from foods and samples associated with food-poisoning outbreaks. For this investigation, the presence of enterotoxin genes, such as those of haemolysin BL, B. cereus enterotoxin T and enterotoxin FM, were assayed by polymerase chain reaction (PCR) methods. Meanwhile, their enterotoxin activities were assayed using the BCET-RPLA kit, haemolytic patterns on sheep blood agar and their cytotoxicity to Chinese hamster ovary (CHO) cells. Results showed that there were 12 enterotoxigenic profiles for the 98 B. cereus group strains collected. In addition, if any of the three types of enterotoxins was present in the B. cereus group cells, these cells were shown to be cytotoxic to the CHO cells. Similar enterotoxigenic profiles could be found among strains of B. cereus, B. mycoides and B. thuringiensis. Thus, all B. cereus group strains may be potentially toxigenic and the detection of these cells in foods is important. We thus designed PCR primers, termed Ph1/Ph2, from the sphingomyelinase gene of B. cereus cells. These primers were specific for all B. cereus group strains and could be used for the detection of B. cereus cells contaminated in food samples.

Animals↗

The new design of an infrared-controlled human-computer interface for the disabled.

This paper reports on the development of an eyeglass- type infrared (IR)-controlled computer interface for the disabled. This system may serve to assist those who suffer from spinal cord injuries or other handicaps to operate a computer. This system is comprised of three major components: 1) an infrared transmitting module, 2) an infrared receiving/signal-processing module, and 3) a main controller, the Intel-8951 microprocessor. The infrared transmitting module utilizes tongue-touch circuitry which is converted to an infrared beam and a low power laser (<0.1 mW) beam. The infrared receiving/signal-processing module, receives the infrared beam and fine tunes the unstable infrared beam into standard pulses which are used as control signals. The main controller is responsible for detecting the input signals from the infrared receiving/signal-processing module and verifying these signals with the mapping table in its memory. After the signal is verified, it is released to control the keys of the computer keyboard and mouse interface. This design concept was mainly based on the idea that the use of an infrared remote module fastened to the eyeglasses could allow the convenient control of the input motion on the keys of a computer keyboard and mouse which are all modified with infrared receiving/signal-processing modules. The system is designed for individuals with spinal cord injuries and disabled in which the subjects' movement are severely restricted. The infrared transmitting module can be easily mounted on eyeglasses or artificial limbs.

Adult↗

Nef-induced CD4 and major histocompatibility complex class I (MHC-I) down-regulation are governed by distinct determinants: N-terminal alpha helix and proline repeat of Nef selectively regulate MHC-I trafficking.

The Nef protein of primate lentiviruses triggers the accelerated endocytosis of CD4 and of class I major histocompatibility complex (MHC-I), thereby down-modulating the cell surface expression of these receptors. Nef acts as a connector between the CD4 cytoplasmic tail and intracellular sorting pathways both in the Golgi and at the plasma membrane, triggering the de novo formation of CD4-specific clathrin-coated pits (CCP). The downstream partners of Nef in this event are the adapter protein complex (AP) of CCP and possibly a subunit of the vacuolar ATPase. Whether Nef-induced MHC-I down-regulation stems from a similar mechanism is unknown. By comparing human immunodeficiency virus type 1 (HIV-1) Nef mutants for their ability to affect either CD4 or MHC-I expression, both in transient-transfection assays and in the context of HIV-1 infection, it was determined that Nef-induced CD4 and MHC-I down-regulation constitute genetically and functionally separate properties. Mutations affecting only CD4 regulation mapped to residues previously shown to mediate the binding of Nef to this receptor, such as W57 and L58, as well as to an AP-recruiting dileucine motif and to an acidic dipeptide in the C-terminal region of the protein. In contrast, mutation of residues in an alpha-helical region in the proximal portion of Nef and amino acid substitutions in a proline-based SH3 domain-binding motif selectively affected MHC-I down-modulation. Although both the N-terminal alpha-helix and the proline-rich region of Nef have been implicated in recruiting Src family protein kinases, the inhibitor herbimycin A did not block MHC-I down-regulation, suggesting that the latter process is not mediated through an activation of this family of tyrosine kinases.

CD4 Antigens↗

Inhibition of NO synthesis or endothelium removal reveals a vasoconstrictor effect of insulin on isolated arterioles.

In this study we tested the hypothesis that insulin may differentially affect isolated arterioles from red (RGM) and white gastrocnemius muscles (WGM) because of their differences in function and metabolic profile. We also determined whether the responses of these arterioles are endothelium dependent and mediated by either prostaglandins or nitric oxide (NO). Arterioles were isolated, pressurized to 85 mmHg, equilibrated in Krebs bicarbonate-buffered solution (pH 7.4) gassed with 10% O2 (5% CO2-85% N2), and studied in a no-flow state. Control diameters for first-order arterioles from RGM averaged 77 +/- 8 micrometers and from WGM averaged 77 +/- 5 micrometers. Cumulative dose-response curves to insulin (10 microU/ml, 100 microU/ml, 1 mU/ml, and 10 mU/ml) were obtained in arterioles before and after endothelium removal or administration of either indomethacin (Indo, 10(-5) M) or NG-nitro-L-arginine (L-NNA, 10(-4) M). Insulin evoked concentration-dependent increases in control diameter of intact RGM and WGM arterioles of 6-26% and 9-28%, respectively. Indo was without any effect on insulin-induced dilation in RGM and WGM arterioles. Insulin-evoked dilation in both RGM and WGM arterioles was completely inhibited and converted to vasoconstriction by endothelium removal and administration of L-NNA. These results indicate that in endothelium-intact arterioles from RGM and WGM, insulin evokes an endothelium-dependent dilation that is equivalent and mediated by NO. In contrast, in the absence of a functional endothelium, insulin evokes arteriolar constriction. The finding that insulin can constrict arterioles, at physiological concentrations, suggests that insulin may play a more significant role in the regulation of vascular tone and total peripheral resistance than previously appreciated.

Animals↗

Telescopic video microlaryngeal surgery.

Although traditional microscopic laryngeal surgery has been well developed, we still may encounter some clinical problems, including exposure difficulty, blind areas during examination, and inadvertent cutting through lesions. These problems may lead to an increased risk of complications or protracted or even repeated operations. We have partially overcome these problems since October 1992 with a new telescopic endolaryngeal surgery technique. This technique was developed by combining and modifying the principles of bronchoscopic surgery and endoscopic sinus surgery. With this technique, it is not necessary to use a very slim laryngoscope or apply overpressure on the larynx when introducing and suspending the laryngoscope. Difficult-to-examine regions can be accessed more easily. We can also get a 3-dimensional impression of lesions and related structures, so it is easier to determine a proper cutting line to avoid cutting through lesions. We consider this telescopic method to be a useful alternative to conventional microscopic laryngeal surgery.

Adult↗