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Biomedical subjects

M H Chen

Publications and source records attributed to M H Chen.

At least 127 records · Page 7Linked to original sources
PubMed1995

Homology modeling of an immunoglobulin-like domain in the Saccharomyces cerevisiae adhesion protein alpha-agglutinin.

The Saccharomyces cerevisiae adhesion protein alpha-agglutinin is expressed by cells of alpha mating type. On the basis of sequence similarities, alpha-agglutinin has been proposed to contain variable-type immunoglobulin-like (IgV) domains. The low level of sequence similarity to IgV domains of known structure made homology modeling using standard sequence-based alignment algorithms impossible. We have therefore developed a secondary structure-based method that allowed homology modeling of alpha-aggulutinin domain III, the domain most similar to IgV domains. The model was assessed and where necessary refined to accommodate information obtained by biochemical and molecular genetic approaches, including the positions of a disulfide bond, glycosylation sites, and proteolytic sites. The model successfully predicted surface exposure of glycosylation and proteolytic sites, as well as identifying residues essential for binding activity. One side of the domain was predicted to be covered by carbohydrate residues. Surface accessibility and volume packing analyses showed that the regions of the model that have greatest sequence dissimilarity from the IgV consensus sequence are poorly structured in the biophysical sense. Nonetheless, the utility of the model suggests that these alignment and testing techniques should be of general use for building and testing of models of proteins that share limited sequence similarity with known structures.

Agglutinins↗
PubMed1995

Comparative cutaneous testing with purified protein derivative and the antigen complex A60 in vaccinated subjects and tuberculosis patients.

Some 840 bacille Calmette-Guérin (BCG)-vaccinated healthy controls and tuberculosis patients from two Chinese hospitals were submitted to comparative skin tests with purified protein derivative of tuberculin (PPD; as reference) and with the antigen complex A60 from Mycobacterium bovis BCG. In a first trial, including 581 persons (185 healthy juveniles, 180 healthy adults and 216 tuberculosis patients), a limited dose of A60 (1 microgram) was used. Performance of the A60 test was similar to that of 5 I.U. PPD for controls (cut-off values = 5 mm induration diameter), but lower than that seen for tuberculosis patients (10 mm cut-off values). A second survey was conducted on 259 persons (109 recently revaccinated healthy persons, considered as tuberculin-negative in the first trial, and 150 tuberculosis patients), using a higher dose of A60 (2 micrograms) and the same dose of PPD (5 I.U.). Similar results were obtained with the two tests in all cases, thus supporting the possibility of PPD replacement by A60 in cutaneous testing. The pattern of induration diameter distribution in healthy subjects who took part in the first testing round (64% positively rate) was displaced to the inactivity side (with a peak at 5 to 9-mm diameter), in comparison with the second round (90% positivity rate and peak at 10-14 mm). This indicates a progressive fading of cellular immunity reactions after BCG vaccination. In tuberculosis patients, no correlation was found among the following three parameters: positivity at cutaneous testing (with PPD or A60), titer of anti-A60 mycobacterial immunoglobulins in blood (IgG titer higher than cut-off line) and presence of mycobacteria in sputum.

Adolescent↗
PubMed1995

Laparoscopic-assisted abdominal aortic aneurysm repair.

Since the advent of laparoscopy, the sweeping changes seen in general surgery have not been paralleled in vascular surgery. There have been case reports of laparoscopic-assisted aortobifemoral bypass for occlusive disease. Because aneurysmal disease comprises the majority of aortic surgery, we pursued animal and cadaveric feasibility studies for laparoscopic-assisted abdominal aortic aneurysm (AAA) repair. We present a case report of the first clinical case performed under Institutional Review Board protocol using this technique. The patient was a 62-year-old male with a 6-cm infrarenal AAA. After obtaining a pneumoperitoneum, a modified fish retractor was used to exclude the bowel. Ten 11-mm ports provided access to laparoscopically dissect the neck of the aneurysm and the iliac vessels. Then, a 10-cm minilaparotomy was performed and standard vascular clamps were inserted via the port incisions. Standard aneurysmorraphy was performed with a polytetrafluoroethylene (PTFE) tube graft. Laparoscopy conferred three major benefits: better visualization of the aneurysm neck, less bowel manipulation, and avoidance of hypothermia. This case report illustrates the feasibility of laparoscopic-assisted aneurysm repair. Controlled human studies will define the role of laparoscopy in AAA surgery.

Aortic Aneurysm, Abdominal↗
PubMed1995

Intracardiac ultrasonographic assessment of atrial septal defect area: in vitro validation and technical considerations.

Assessment of atrial septal defect (ASD) size and shape is important for planning and guiding its transcatheter occlusion and can potentially be achieved by intracardiac ultrasonography (ICUS). ICUS accuracy, however, must first be established against stable standards and technical imaging requirements defined. We therefore used 10, 20, and 30 MHz ICUS catheters to examine 17 ASDs that were 0.16 to 6.7 cm2 in area and were surgically created in excised ovine hearts with 10, 20, and 30 MHz ICUS catheters. ASD shape and area by ICUS were compared with direct video images of the actual ASD. In all instances minimal area by ICUS pullback agreed well with actual values (y = 1.04x + 0.2, SEE = 0.23 cm2, r = 0.99) and corresponded well with defect shapes. The maximum angle between ultrasonography beam and septal plane allowing for complete ASD visualization was 20 degrees. The angle depended on transducer frequency and septal thickness. This new technique has potential value for the accurate assessment of ASD shape and size and may be especially useful in the setting of transcatheter occlusion.

Animals↗
PubMed1995

Helicobacter pylori associated with a high prevalence of duodenal ulcer disease and a low prevalence of gastric cancer in a developing nation.

This study examines the relationship between Helicobacter pylori infection and peptic ulcer disease and gastric cancer--in particular, the presence or absence of bacteria, the grading of gastritis, and the degree of inflammation in the antral and oxyntic mucosae. The grading of gastritis and the detection of H pylori were determined by histology using the Sydney system. Of the 1006 patients examined, 34.5% had duodenal ulcer disease, 3.5% gastric ulcer disease, and 2% with coexistent ulceration. Most patients (50.2%) were classified as having non-ulcer dyspepsia. Altogether 2.4% of patients had gastric cancer and two further patients had carcinoma in the gastric stump. Of the ulcer disease patients, 87.2% had histological evidence of H pylori infection. After patients who had taken antibiotics or bismuth compounds in the preceding four weeks were excluded, 98.9% of the duodenal ulcer disease, 100% of the gastric ulcer disease, and 100% of the coexistent ulcer disease patients had evidence of H pylori infection. In patients with gastric cancer who had not taken antimicrobial agents in the four weeks before endoscopy, 83.3% had evidence of H pylori infection. Thus, there was a high rate of duodenal ulcer disease and a low rate of gastric ulcer disease in southern China, an area of low gastric cancer mortality. There was a specific topographical relationship between H pylori, the histological response, and gastroduodenal disease. Our data suggest that the status of a nation as either 'developed' or 'developing' can not be used to predict the upper gastrointestinal disease profile of its population.

Adolescent↗
PubMed1995

Pharmacological and morphological effects of in vitro transluminal balloon angioplasty on normal and vasospastic canine basilar arteries.

Despite growing clinical use of transluminal balloon angioplasty (TBA) to treat cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH), the precise mechanism of action of balloon dilation on the cerebral arterial wall is unknown. In this experiment the authors examined the pharmacological and morphological changes in 10 normal and 12 vasospastic canine basilar arteries following in vitro silicone microballoon TBA. For the SAH group in which the double-hemorrhage model was used, vasospasm was confirmed by angiography and the animals were killed on Day 7 after the first SAH. In vitro TBA was performed on basilar arteries from normal and SAH dogs immediately after sacrifice and removal of the brain. The procedure was performed while the arteries were maintained in oxygenated Krebs buffer. In the pharmacological studies, potassium chloride, prostaglandin F2 alpha, serotonin, and noradrenaline were used as vasoconstrictors, and bradykinin and calcium ionophore A23187 were used to produce an endothelium-dependent dilation. In both normal and vasospastic groups, the pharmacological responses of dilated segments of basilar arteries were compared to those of nondilated segments of the same arteries. Vessels from all groups were examined using scanning electron microscopy (EM) and transmission EM. Scanning EM was used to study the intact vessel wall, the smooth-muscle cell layer obtained after digestion with hydrochloric acid, and the extracellular matrix obtained after digestion with bleach. Cross-sections of the vessel wall were examined using transmission EM. The most striking finding was that immediately after in vitro TBA of both normal and vasospastic canine basilar arteries, there was a significant reduction (p < 0.05) of responses to both vasoconstrictors and vasorelaxants. As revealed by scanning EM and transmission EM, both normal and vasospastic vessels dilated with TBA showed flattening and patchy denudation of the endothelium, and straightening and occasional rupturing of the internal elastic lamina. In addition, vasospastic vessels dilated with TBA showed decreased surface rippling and mild stretching and straightening of smooth-muscle cells, and mild thinning of the tunica media. There was no gross vascular disruption or obvious change in the extracellular matrix of the vessel walls of either normal or vasospastic arteries after TBA. These results suggest that functional impairment of vasoreactivity in the vessel wall as a result of mechanical stretching of the smooth-muscle layer plays a more important role than structural alteration, at least in the immediate dilation produced in vasospastic arteries by TBA.

Analysis of Variance↗
PubMed1995

The effect of cobalt-60 irradiation on bone marrow cellularity and alveolar osteoclasts.

The purpose of this study was to investigate the mechanisms of bone resorption in the rat dentoalveolar complex occurring as a result of orthodontic appliance therapy. Utilizing whole body radioactive cobalt 60 (Co60) irradiation in combination with orthodontic tooth movement, the activation, recruitment, and life span of osteoclasts was studied. Thirty-four adult Fischer 344 rats were irradiated with 10 and 20 Gray of Co60. Twelve days after irradiation, each rat was fitted with an orthodontic appliance; the rats were sacrificed 7, 14, or 21 days after appliance placement. To serve as controls, another group of 12 rats was subjected to orthodontic treatment only. Histologic sections were prepared from decalcified maxillary alveolar process, and osteoclasts were counted. In the control group, osteoclasts were presence in the periodontal membrane for four weeks after appliance placement, and the largest number of osteoclasts was observed in the second week. The smaller total irradiation dose (10 Gray, administered in 5 daily fractions) induced a transient reduction in the bone marrow cell count of more than 90%, followed by a complete rebound. The peak osteoclst number also was increased 110%. In contrast, the higher total dose (20 Gray, administered in 10 daily fractions) resulted in a reduction of 60% in bone marrow cellularity and a decrease in the peak osteoclast number by 40%. Fluctuations in bone marrow cellularity generally corresponded to similar variations in the osteoclast number, but there was an apparent lack of correspondence between bone marrow cell and white blood cell values. The total period of osteoclastic presence in the periodontal membrane following orthodontic activation, normally four weeks in duration, was reduced by one week in the irradiated animals. These findings lead us to speculate that the mechanisms of osteoclast activation and recruitment following orthodontic appliance therapy may involve three consecutive waves of osteoclast maturation. The total duration of osteoclastic bone resorption lasts 4 weeks, but the osteoclast lie span is calculated to be 9 to 10 days (9 days x 3 waves = 27 days). If the results of this study are proved to be reproducible, a scientific basis may have been provided to support the common practice in orthodontics wherein the duration between orthodontic appointments is 4-5 weeks.

Alveolar Process↗
PubMed1995

Gastric atrophy and regional variation in upper gastrointestinal disease.

OBJECTIVE: To compare the upper gastrointestinal disease profiles of Helicobacter pylori-infected patients drawn from two distinct regions of China, one with a low incidence of gastric cancer (Guangzhou) and the other with a high incidence of gastric cancer (Lanzhou). The age-standardized prevalence of H. pylori within the populations of these two cities was similar (approximately 56%). In these patient groups, the prevalence of different gastrointestinal disease states and the occurrence and severity of gastritis, gastric atrophy, and intestinal metaplasia were compared. METHODS: This study was based on consecutive patients: 265 from Guangzhou and 275 from Lanzhou. The grading of gastritis and the detection of H. pylori was determined by histology using the "Sydney System". RESULTS: The ratio of cases of duodenal to gastric ulceration for the two cities was: Guangzhou 14:1 and Lanzhou 3:1. Gastric cancer was more prevalent in the patients from Lanzhou (9.8%) than from Guangzhou (3%). In nonulcer dyspepsia patients from Guangzhou, gastritis was predominantly antral, whereas, in Lanzhou, gastritis was predominantly uniform. The amount and severity of atrophy was significantly greater in Lanzhou compared with Guangzhou. CONCLUSIONS: The prevalence of atrophy, rather than age of acquisition and prevalence of H. pylori infection, appears to be a marker of the major upper gastrointestinal disease profiles of a region. Development of atrophy, although apparently related to H. pylori infection, may be a multifactorial condition. Differences in diet, as seen between Guangzhou and Lanzhou, may be important in this regard. Understanding the factors leading to the development of atrophy may enhance our understanding of processes leading to gastric malignancy.

Atrophy↗
PubMed1994

A unique mitochondria-associated membrane fraction from rat liver has a high capacity for lipid synthesis and contains pre-Golgi secretory proteins including nascent lipoproteins.

An endoplasmic reticulum-like membrane fraction, termed the "mitochondria-associated membrane" (MAM), that co-isolates with mitochondria from rat liver has been characterized. One potential function of the MAM is as a membrane bridge between the endoplasmic reticulum and mitochondria that may be involved in transfer of phospholipids between these two organelles (Vance, J. E. (1990) J. Biol. Chem. 265, 7248-7256). A polyclonal antibody directed against a peptide corresponding to the C terminus of phosphatidylethanolamine N-methyltransferase-2, a specific marker protein of the MAM (Cui, Z., Vance, J. E., Chen, M. H., Voelker, D. R., and Vance, D. E. (1993) J. Biol. Chem. 268, 16655-16663), was used in immunofluorescence and immunogold electron microscopy localization studies in rat hepatocytes. Immunoreactive protein was clustered in regions of the cell that did not correspond to the bulk of endoplasmic reticulum. A second potential role for the MAM, as a component of the secretory pathway that supplies lipids for assembly into very low density lipoproteins, has been examined. The MAM contains enzymes of similar, or higher, specific activities to those enzymes in the endoplasmic reticulum for the synthesis of phospholipids, triacylglycerols, cholesterol, and cholesteryl esters. Specific activities of diacylglycerol acyltransferase, acyl-coenzyme A:cholesterol acyltransferase, and phosphatidylserine synthase (base exchange enzyme) are enriched 2.2-3.4-fold in the MAM compared with endoplasmic reticulum. In addition, the microsomal triacylglycerol transfer protein, which is required for the assembly/secretion of apolipoprotein B-containing lipoproteins, was present in the MAM. Nascent apolipoprotein B-containing lipoproteins were isolated from the lumen of the MAM. These lipoproteins had the same average density and composition as nascent apolipoprotein B-containing lipoproteins isolated from heavy and light endoplasmic reticulum fractions, from the Golgi, and lipoproteins newly secreted by cultured rat hepatocytes. The MAM is a pre-Golgi compartment of the secretory route, as shown by pulse-chase studies with apolipoprotein B and albumin, as well as the sensitivity of luminal apolipoprotein B to endoglycosidase H.

Animals↗
PubMed1994

Expression of alpha-amylases, carbohydrate metabolism, and autophagy in cultured rice cells is coordinately regulated by sugar nutrient.

A rice suspension cell culture system has been established to study how sugar depletion regulates alpha-amylase expression, carbohydrate metabolism, and other physiological and cellular changes. It is shown here that a group of 44 kDa alpha-amylases are constitutively expressed whether or not the cells are starved of sucrose. However, expression of a new group of alpha-amylases of 46 kDa is dramatically induced when cells are starved of sucrose. Cellular sugar and starch were rapidly consumed and metabolic activity was decreased in the starved cells. Extensive autophagy also occurred in the starved cells, which caused an increase in vacuolar volume and degradation of cytoplasmic constituents including amyloplasts. Immunocytochemical studies revealed that alpha-amylases are localized in starch granules within amyloplasts, in cell walls, and in some of the vacuoles. The presence of putative signal sequences in the N-termini of nine rice alpha-amylases suggests hitherto unidentified pathways for import of alpha-amylases into amyloplasts. The studies show that differential alpha-amylase expression, carbohydrate metabolism, metabolic activity, and vacuolar autophagy are coordinately regulated by the sugar level in the medium. As the starved suspension cells exhibit some sugar-regulated characteristics of alpha-amylase expression in germinating rice embryos as well as physiological changes similar to those in senescing cells, this system represents an ideal tool for studying cellular, biochemical, and molecular biological aspects of alpha-amylase gene regulation, carbohydrate metabolism, senescence, and protein targeting in plants.

Amino Acid Sequence↗
PubMed1994

Serological analysis of pulmonary and extrapulmonary tuberculosis with enzyme-linked immunosorbent assays for anti-A60 immunoglobulins.

IgA, IgG, and IgM antibodies to mycobacterial antigen A60 were measured by ELISA in blood, pleural fluid, and cerebrospinal fluid from 560 patients with pulmonary and/or extrapulmonary tuberculosis who were being treated at hospitals in northern China and from 734 uninfected controls. Among 529 healthy persons (most of whom had been vaccinated with bacille Calmette-Guérin [BCG] and 287 of whom were tuberculin-positive), the rate of false-positive results was negligible; this observation ruled out interference of remote BCG vaccination with A60 assays at the chosen cutoff level. Rates of positivity for IgM and IgG, respectively, were 80% and 36% among patients with active primary pulmonary tuberculosis, 31% and 88.5% among patients with active postprimary pulmonary tuberculosis, 0 and 41% among patients with inactive pulmonary tuberculosis, and 30%-61% and 69%-86% among patients with extrapulmonary tuberculosis. Paired samples of blood and pleural fluid from patients with pleurisy contained IgA antibody to A60 at equal titers; in contrast, most patients with tuberculous meningitis (100% of whom had a positive ELISA result) had higher levels of IgG antibody to A60 in cerebrospinal fluid than in blood--proof of intrathecal synthesis.

Adult↗
PubMed1994

Intracardiac ultrasound measurement of volumes and ejection fraction in normal, infarcted, and aneurysmal left ventricles using a 10-MHz ultrasound catheter.

BACKGROUND: Our objective was to examine the accuracy of intracardiac ultrasound (ICUS) measurement of left ventricular (LV) volumes and ejection fraction (EF) using a 10-MHz ultrasound catheter. ICUS can image the LV in cross sections at all levels along the long axis with a transducer mounted on the tip of a catheter. Sequential serial LV cross-sectional images can be obtained during cardiac catheterization and used to calculate LV volumes by Simpson's rule. This technique may be an alternative to contrast LV angiography. METHODS AND RESULTS: A beating-heart in vivo model was created to measure LV volume directly and continuously with an intracavity high-compliance latex balloon connected to a calibrated extracardiac reservoir in eight dogs in 35 experimental stages. A 10F ICUS catheter with a 10-MHz single-element transducer was introduced retrogradely via the aortic valve to the apex. Series of sequential LV cross-sectional images were recorded from the apex to the base during a calibrated pullback of the catheter. At each 5-mm interval, the LV cross section was traced at end diastole and end systole. LV volume was calculated by Simpson's rule by integrating all segmental areas multiplied by segmental height. The effect on accuracy of selecting 5-, 10-, or 15-mm heights or a single section at the midventricular level for measurement was assessed. The influence of distorted ventricular shape on the accuracy of ICUS measurements of LV volume was evaluated. This method was applied in 19 experimental stages in 10 intact dogs and pigs catheterized via the femoral artery. In the in vivo canine model, LV end-diastolic volume, end-systolic volume, and EF determined by ICUS using 5-, 10-, or 15-mm segments were not different from the actual measurements. But correlation and agreement between ICUS end-diastolic volume and direct measurements for 5- and 10-mm segments were significantly better than for 15-mm segments or a single section. Similar excellent correlations and agreement were observed for actual and ICUS-derived end-systolic volumes using 5-, 10-, or 15-mm segments. The ICUS-derived EF correlated very well with actual EF with a small measurement error of 3.91 +/- 2.59% for 5-mm or 4.13 +/- 2.79% for 10-mm segments but a significantly greater measurement error for 15-mm segments (5.35 +/- 3.76%) or single sections (14.8 +/- 12.2%). The presence of LV infarction or aneurysm did not significantly influence the accuracy of ICUS calculations for segmental heights < or = 10 mm. Application in intact animals demonstrated a good correlation between stroke volume measured by ICUS and by thermodilution or flowmeter. ICUS-derived LV volumes correlated well with biplane angiographic volumes, with a tendency toward underestimation. There was no significant difference between ICUS-determined LV EF and EF determined by angiography. CONCLUSIONS: Intracardiac echocardiography accurately measures LV volumes and global systolic function in both regularly shaped and distorted left ventricles. This technique directly and continuously visualizes circumferential LV endocardium and wall thickness without contrast agents or geometric assumptions for calculation of LV volume. Thus, it should be particularly useful in patients at high risk for contrast-related complications or distorted LV shapes in which geometric assumptions may not be valid.

Animals↗
PubMed1994

[The clinical significance of detection of anti-BCG-A60 antibody by ELISA in diagnosis of tuberculosis].

In order to find out the clinical significance of detection of BCG-A60 antibody by ELISA in diagnosis of tuberculosis, 734 cases of non-tuberculosis patients and 560 cases of tuberculosis has been examined. The result of A60-ELISA test for IgG as diagnosilic criteria of tuberculosis, sensitivity is 84.5%, specificity is 89.3%, It shows satisfactory effect, so it can be used as an helpful method in clinical diagnosis of tuberculosis. IgG, IgA and IgM are measured at same time, and find that a combined use of the IgG and IgM ELISA studies considerably increases the significance of the test.

Antibodies, Bacterial↗
PubMed1994

Transcutaneous ultrasound of the cervical esophagus in patients with esophageal carcinoma.

Transcutaneous ultrasound of the cervical esophagus was performed in 46 patients with esophageal carcinoma and in 35 controls. The former had 24 upper segmental lesions and 22 lower segmental lesions. The level of the sternoclavicular joint was used to divide the esophagus into the upper segmental (USE) and lower segmental esophagus (LSE). The anterior esophageal wall thickness and luminal dimensions were measured before and immediately after phonation. The mean wall thickness in the controls was 1.8 mm before phonation and 2.1 mm after phonation, with a significant difference (t test, P < 0.05). The mean wall thickness in the USE carcinoma group was 4.3 mm and 4.4 mm before and after phonation respectively. There was a significant difference between the controls and USE carcinoma groups (t test, P < 0.05). The cross sectional area, which was calculated as the product of anterio-posterior and lateral diameters, averaged 28 mm2 before phonation in the controls and increased to 44 mm2 after phonation (t test, P < 0.05). The mean cross sectional area in the USE carcinoma group was significantly smaller before phonation than that in the controls (t test, P < 0.05) and showed little change after phonation. No significant difference was found between LSE carcinoma and control groups. Transcutaneous ultrasound appears to be a promising non-invasive method of investigation for cervical esophageal carcinoma.

Adult↗
PubMed1994

[Manifestation of abdominal segment of esophagus by body surface ultrasonography and determination of normal values].

Scanning over the oblique section of xiphoid-left costal margin by body surface ultrasonography was adopted in the present study. The esophageal hiatus showed triangle-like strong echo which was called by the author "Hom Dill Casoue". Its dorsal aspect could be confirmed as the abdominal segment of the esophagus. In a general survey of 268 cases of high risk crowd in high incidence area, 82% (217 cases) were manifested clearly. The mean wall thickness was 2.58 +/- 0.54mm, and the mean length 23.7 +/- 5.6mm. Among the 9 cases suspicious of increased thickness, 5 were confirmed to have esophageal lesions. Among them there were 2 cases of esophageal carcinoma, and 2 cases of inflammatory ulcer with edema, and one case of hiatal hernia. The accuracy rate of diagnosis for those with clear manifestation by ultrasonography reached as high as 98% (218/222 cases). This method provides a new measure for the general survey of esophageal diseases in the abdominal segment.

Adult↗
PubMed1994

A non-excluded set method for the calculation of paternity probability.

Forensic judgment of paternity depends on genetic and nongenetic evidence. Sometimes genetic marker tests can provide clear evidence to exclude a falsely accused man, but they do not always succeed. On the other hand, if the accusation is true, the alleged father will not ever be excluded by genetic marker tests. When a nonexclusion case occurs after one or more marker tests, a report of paternity probability is required in the court. The current methods for calculating the paternity probability are the "paternity index method" and the "nonexclusion method." A number of recent articles have openly debated the fallacies, validity and utility of both methods. This paper briefly reviews the two methods and proposes a non-excluded set method along with examples for illustrating the various patterns of paternity probabilities of the three methods.

Forensic Medicine↗