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Biomedical subjects

C H Wang

Publications and source records attributed to C H Wang.

At least 163 records · Page 9Linked to original sources

Leucine-2-alanine enkephalin induced delta opioid receptors internalization expressed stably in CHO cells.

AIM: To characterize the internalization of delta opioid receptors (DOR) stably expressed in Chinese hamster ovary (CHO) cells and the role of the C-terminal in this process. METHODS: Receptor membrane anchoring was shown by immunofluorescence microscopy. Receptor internalization was assessed by measuring the radioligand binding resistant to the acid-buffer wash. RESULTS: Originally, all the wild-type (CHO-W) and C-truncated (CHO-T) DOR expressed were localized to the membrane. Agonist [3H] leucine-2-alanine enkephalin (LAE) but not the antagonist [3H]diprenorphine (Dip) induced rapid receptor internalization. The internalization of C-truncated DOR in CHO-T was similar to that of the wild-type in maximal level, but climbed up more slowly. DOR internalization was extracellular osmolarity- and temperature-sensitive. Pertussis toxin and universal protein kinase inhibitor staurosporine had no effect on it. CONCLUSION: DOR internalization is an agonist and clathrin-coated pits dependent, but post-receptor cellular signal transduction independent process; moreover, the C-terminal of DOR, not engaged in membrane anchoring, affects the initialization of DOR internalization.

Animals↗

Myocardial effects of beta-agonist stimulation in rats with chronic left ventricular dysfunction treated with an angiotensin-converting enzyme inhibitor.

BACKGROUND: This study measured morphological and hemodynamic changes and renin-angiotensin responsiveness of the left ventricle (LV) to beta-agonist stimulation in a Sprague-Dawley rat model of myocardial dysfunction produced by coronary artery ligation. METHODS: The LV function and papillary muscle mechanics were measured after 12 weeks of captopril treatment (2 g/l in drinking water) following left coronary artery ligation or a sham operation. Fifty-two rats were divided into three groups: those with sham operations, those with small infarcts (infarct size [IS] < 30% LV) and those with large infarcts (IS > or = 30% LV). RESULTS: The results showed that LV end-diastolic pressures were elevated in the large-infarct group regardless of treatment with the angiotensin-converting enzyme inhibitor (ACEI), and the LV weight was reduced in the ACEI-treated rats. In addition, the uninfarcted LV posterior papillary muscle of the large-infarct rats showed an impaired response to isoproterenol stimulation, including the developed tension, positive and negative rate of tension development, time to peak tension, and time to half relaxation. CONCLUSION: Chronic captopril treatment improved isoproterenol-stimulated muscle isometric function in rats following myocardial infarction, possibly through the beta-receptor pathway.

Adrenergic beta-Agonists↗

[Ciliary neurotrophic factor prevents NO mediated cytotoxicity in cultured hippocampal neurons].

The effects of ciliary neurotrophic factor on nitric oxide (NO)-induced cytotoxicity were examined in primary cultures of rat hippocampal neurons. The neurotoxicity was induced by NO donors, such as sodium nitroprusside (SNP), s-nitroso-N-acetylpenicillamine (SNAP), NOS substrate, L-arginine (L-Arg) and ionomycin (a calcium ionophore). Preincubation of the cultures with ciliary neurotrophic factor (CNTF) for 24 h promoted neuronal survival and decreased the LDH efflux and the level of NO induced by ionomycin. The protective effects of CNTF against NO cytotoxicity were concentration-dependent. The above findings suggest that CNTF promotes the survival of neurons in the hippocampus by preventing NO mediated neurotoxicity.

Animals↗

[Protection of spinal motorneurons of section sciatic nerve by transplantation of NT-4 genetically expression cells into the side of section].

The NT-4 genetically engineered cells were made by infecting L-6TG cell (a rat myoblast cell line) in vitro with a retroviral vector pN2A containing the rat NT-4 cDNA. The bioactivities were determined by bioassay of PC12 cell survival rate. The rat with left sciatic nerve transaction was used as a model for treatment by implanting NT-4 genetically modified cell. The condition of motorneurons was assessed by Nissl stain and ChE stain. The results showed that: (1) the percentage of surviving Nissl-stained neurons on the lessened side of NT-4 (+) grafts significantly increased as compared to that with NT-4 (-) grafts 2-3 weeks and 3 months after sciatic nerve transaction, and (2) the grafted cells produced significant increase in the positive ChE stained area after sciatic nerve transaction in 1-3 weeks. Our observations indicate that adult motor neurons are still able to respond to neurotrophic factors and they may require the factors for survival.

Animals↗

[Relationship between the expression of p53 and C-erbB-2 oncoproteins and biologic behaviors of human mucoepidermal carcinoma of salivary gland]

OBJECTIVE:To study the relationship between the expression of p53 and c-erbB-2 proteins and biologic behaviors of human mucoepidermal carcinoma.METHODS:The expressions of oncogene proteins p53 and c-erbB-2 in 32 cases of human mucoepidermal carcinomas were investigated with microwave immunohistochemistry.RESULTS:p53 and c-erbB-2 oncoproteins in duct epithelial cells around cancer tissues were 10.0% and 15.0%,respectively,but those in the acinar cells affiliated with the salivary gland ducts were negative. The positive rates of p53 oncoprotein and c-erbB-2 oncoprotein in mucoepidermal carcinoma were 40.6% and 46.9%,respectively. Both p53 and c-erbB-2 oncoproteins were expressed in mucous cells,epidermoid cells and intermediate cells of mucoepidermal carcinoma. The expression of p53 and c-erbB-2 oncoproteins in human mucoepidermal carcinoma tissues were related to histological patterns,differentiation degree of cancer and tumor recurrence (P<0.01).CONCLUSION:This study suggested that both p53 and c-erbB-2 on coproteins,as the markers for differentiation of cancer,might be useful in screening progress and evaluating prognosis of patients with mucoepidermal carcinoma.

Journal Article↗

Diagnostic value of dobutamine echocardiography in patients with angina-like symptoms preceding syncope.

Dobutamine echocardiography was performed in 55 patients with syncope which was clinically suspected to be angina-related. We evaluated the value of using a single test, dobutamine echocardiography, in differentiating real ischemia-related from vasovagal syncope which was diagnosed by a tilt test. During testing, supraventricular arrhythmia was provoked in four (7.2%) patients. Dobutamine echocardiography identified all of six (10.9%) patients (sensitivity 100%), who were found with significant coronary stenosis by coronary angiograms. The etiology of syncope in the remaining 45 patients was investigated further by tilt testing, the findings of hypotension and bradycardia during which were compared head to head with those of dobutamine echocardiography. Tilt testing diagnosed vasovagal syncope in 31 patients, in whom only 19 (61.3%) patients developed vasovagal reflex during dobutamine echocardiography. Conclusively, dobutamine echocardiography had a high sensitivity in identifying syncope related to myocardial ischemia in patients with coronary stenosis, but a low sensitivity (61.3%), high specificity (90.5%) and high positive predictive value (81.8%) in detecting the syncope patients with angina caused by vasovagal effect.

Aged↗

Early induction and augmentation of parasitic antigen-specific antibody-producing B lymphocytes in the non-Peyer's patch region of the small intestine.

In this study, B lymphocytes from the small intestine of immunized rats were examined for their expression of specific antibodies against Trichinella spiralis (TS) antigen. The isotypes of the antigen-specific antibodies on B cells were examined via immunofluorescence microscopy. Monoclonal mouse anti-rat IgE, IgG1, IgG2a, IgG2b, IgG2c, IgA and IgM primary antibodies in conjunction with FITC-conjugated goat anti-mouse Ig secondary antibody and XRITC-conjugated 9D4 T. spiralis antigen were used to study the dynamics of the appearance of activated B lymphocytes in the small intestine, Peyer's patch, both the germinal center (PP-GC) and the non-germinal center (PP-NGC), the mesenteric lymph node (MLN), and the spleen. The results demonstrate that activated B cells are elicited by TS in the non-Peyer's patch region of the small intestine to express all isotypes of antibodies against TS antigen. IgG- and IgE-producing cells (Ab-PC) began proliferation only 1 and 2 days after infection, respectively. The strongest response was mounted by the IgE-PC in the lamina propria of the intestine. The response by IgA-PC generated was not only significantly delayed and also much weaker than that of the IgE- and IgG-PC. Peyer's patches failed to be a significant contributor in this immune response. Although this antigen-specific immune response was produced in the MLN and the spleen, it was weaker than that of the small intestine. The study indicates the potential ability of an immunized host to generate an early, yet effective, humoral immunity against T. spiralis in the non-Peyer's patch region of the small intestine."

Animals↗

Neuroimaging and treatment implications of patients with multiple epidural spinal metastases.

BACKGROUND: Although multiple spinal epidural metastases (MEMs) commonly occur in cancer patients, their clinical significance remains uncertain. The authors attempted to ascertain the incidence of MEMs and their association with the completeness of spinal imaging by magnetic resonance (MR) scanning versus myelography to determine how often they are missed because of incomplete spinal imaging and to assess their prognostic and treatment implications. METHODS: A review of 337 epidural spinal cord compression (ESCC) cases seen at the Mayo Clinic between 1985 and 1993 was conducted. RESULTS: ESCC patients undergoing myelography only were significantly more likely to undergo complete spinal imaging (CSI)than patients undergoing either MR scan only or both imaging modalities (P < 0.0001). MEMs were detected in 32% of patients undergoing CSI and 18% of patients with incomplete spinal imaging (P=0.02). Failure to image the cervical spine in patients with symptomatic thoracic or lumbar epidural lesions would have missed secondary epidural lesions in only 1% of patients; however, this figure increased to 21% for failure to image either the thoracic or lumbosacral spine when symptomatic disease was located elsewhere. Radiation oncologists included secondary epidural deposits in treatment ports in 93% of MEM cases. In a multivariate model, the presence of MEMs was an independent prognostic factor for poorer survival. CONCLUSION: The incidence of MEMs in patients with ESCC is approximately 30%, and their presence frequently alters treatment plans. It appears safe to forgo cervical spine MR scanning in patients with radiographically verified thoracic or lumbar ESCC; however, careful imaging of the thoracic and lumbar spine should be considered in all ESCC patients to detect MEMs.

Adolescent↗

Trends in the incidence of myocardial infarction and in mortality due to coronary heart disease, 1987 to 1994.

BACKGROUND AND METHODS: To clarify the determinants of contemporary trends in mortality from coronary heart disease (CHD), we conducted surveillance of hospital admissions for myocardial infarction and of in-hospital and out-of-hospital deaths due to CHD among 35-to-74-year-old residents of four communities of varying size in the United States (a total of 352,481 persons in 1994). Between 1987 and 1994, we estimate that there were 11,869 hospitalizations for myocardial infarction (on the basis of 8572 hospitalizations sampled) and 3407 fatal coronary events (3023 sampled). RESULTS: The largest average annual decrease in mortality due to CHD occurred among white men (change in mortality, -4.7 percent; 95 percent confidence interval, -2.2 to -7.1 percent), followed by white women (-4.5 percent; 95 percent confidence interval, -0.7 to -8.2 percent), black women (-4.1 percent; 95 percent confidence interval, -10.3 to +2.5 percent), and black men (-2.5 percent; 95 percent confidence interval, -6.9 to +2.2 percent). Overall, in-hospital mortality from CHD fell by 5.1 percent per year, whereas out-of-hospital mortality declined by 3.6 percent per year. There was no evidence of a decline in the incidence of hospitalization for a first myocardial infarction among either men or women; in fact, such hospital admissions increased by 7.4 percent per year (95 percent confidence interval for the change, +0.5 to +14.8 percent) among black women and 2.9 percent per year (95 percent confidence interval, -3.6 to +9.9 percent) among black men. Rates of recurrent myocardial infarction decreased, and survival after myocardial infarction improved. CONCLUSIONS: From 1987 to 1994, we observed a stable or slightly increasing incidence of hospitalization for myocardial infarction. Nevertheless, there were significant annual decreases in mortality from CHD. The decline in mortality in the four communities we studied may be due largely to improvements in the treatment and secondary prevention of myocardial infarction.

Adult↗

Prognostic value of dobutamine echocardiography in patients after Q-wave or non-Q-wave acute myocardial infarction.

We investigated the role of dobutamine echocardiography in predicting future spontaneous events in patients with Q-wave or non-Q-wave first acute myocardial infarction (AMI). DE was performed in 168 patients with a Q-wave AMI and 105 patients with a non-Q-wave AMI. Patients were observed for hard events (cardiac death and nonfatal reinfarction) and all spontaneous events (hard events and unstable angina). When compared to patients with a Q-wave AMI, patients with non-Q-wave AMI had a higher rate of positive dobutamine echocardiographic results (51.8% vs 80.0%, p <0.0001), greater changes in wall motion score index (WMSI) (0.31+/-0.17 vs 0.42+/-0.23, p = 0.001), and more remote zone ischemia (27.9% vs 43.8%, p = 0.0072). Patients with non-Q-wave infarct had a higher all-event rate, but a similar hard-event rate. In patients with a positive dobutamine echocardiogram (DE), the rate of hard or all events was similar, regardless of different infarct patterns. Patients with a negative DE had a higher event-free survival rate for all events in both Q-wave (85.2% vs 60.9%, p <0.0001) and non-Q-wave (76.2% vs 52.4%, p = 0.0083) groups. By stepwise analysis in the Q-wave group, the most important predictors were peak stress WMSI and diabetes for hard events, and a positive DE and baseline WMSI for all events. However, in the non-Q-wave group, the strongest predictors were dobutamine time for hard events and positive DE for all events. We conclude that a positive DE is a powerful predictor of future spontaneous events in patients after either a Q-wave or non-Q-wave AMI. However, for hard events, high-risk patients with different infarct patterns were recognized with variable efficiency by different dobutamine echocardiographic variables.

Aged↗

Characterization of kinetics of anti-Trichinella spiralis newborn larvae immunity in rats.

The kinetics of anti-T. spiralis newborn larvae (NBL) immunity and its dose effects were studied in vivo. Rats were either immunized with newborn larvae i.v. or muscle larvae per os and challenged with newborn larvae either i.v. or i.p. on day 7 up to day 27 after immunization. Immunity was assessed by examining the muscle larvae burden or the larval recovery from the peritoneal cavity. Recovered newborn larvae were further examined for cell adherence and viability. Results indicate that as early as 9 days after infection and only 3 days after newborn larvae production in vivo, specific anti-newborn larvae immunity was developed. Peritoneal cells as well as blood cells adhered to the cuticles of the larvae and killed them. When different doses of immunization were examined, it was found that 2,000 muscle larvae per os induced the strongest immunity as compared to 500, 5,000 or 6,000. Such immunity maintained its strength when challenge infection with newborn larvae reached 50,000 dosage and it declined significantly when the dose reached 100,000. This indicates that the immune cells and antibodies are not re-deployed.

Animals↗

Systemic chemotherapy alone for patients with non-acquired immunodeficiency syndrome-related central nervous system lymphoma: a pilot study of the BOMES protocol.

BACKGROUND: Anecdotal reports have suggested that systemic chemotherapy with agents that better cross the blood-brain barrier may result in long term disease remission in some patients with central nervous system (CNS) lymphoma. This treatment strategy has the advantage of sparing patients the late neurologic complications from brain irradiation. METHODS: Eligible patients were required to 1) have tissue-proven and measurable non-acquired immunodeficiency syndrome (AIDS)- related primary or metastatic CNS lymphoma; 2) have normal hemogram, renal function, and hepatic function; 3) be age < or = 75 years; and 4) have provided informed consent. Patients with lymphoblastic lymphoma or patients who previously had been exposed to nitrosoureas, etoposide, or high dose methotrexate were not eligible. The systemic chemotherapy (BOMES regimen) included carmustine, 65 mg/m2/day, intravenously (i.v.) on Days 1-2; vincristine, 2 mg/day, i.v. on Days 1 and 8; methotrexate, 1.5 g/m2, i.v. on day 15 followed by leucovorin rescue; etoposide, 50 mg/m2/day, i.v. on Days 1-5; and methylprednisolone, 200 mg/day, i.v. on Days 1-7; repeated every 4 weeks (BOMES regimen). Four doses of intrathecal methotrexate were given to patients who had involvement in the cerebrospinal fluid. RESULTS: Between March 1991 and March 1997 a total of 19 patients were enrolled on the study. There were 13 men and 6 women, with a median age of 57 years. Fourteen patients had primary CNS lymphoma and 5 patients had concurrent extra-CNS lymphoma. Nine patients previously had been treated by radiotherapy (four patients), chemotherapy (three patients), or both (two patients). There were 11 complete remissions (CR) (57.9%) and 5 partial remissions (26.3%), with a total remission rate of 84.2%. One patient had had progressive brain lymphoma during systemic chemotherapy with the conventional cyclophosphamide, doxorubicin, vincristine, and prednisolone regimen, but achieved CR soon after the regimen was changed to BOMES. The median time to progression of the responders was 6 months. At last follow-up, 4 patients were alive without lymphoma at 10, 47, 64, and 66 months, respectively. There were two treatment-related deaths due to sepsis. Another two patients died of fulminant hepatitis that most likely was chemotherapy-related reactivation of chronic B viral hepatitis. CONCLUSIONS: The authors believe systemic chemotherapy alone may result in long term disease remission in some select patients with non-AIDS-related CNS lymphoma. Further investigation for better protocols is mandatory.

Adult↗

Identification of a novel missense mutation of the SMN(T) gene in two siblings with spinal muscular atrophy.

Spinal muscular atrophy (SMA) is a motor neuron disease caused by mutations in the telomeric copy of the survival motor neuron (SMN(T)) gene. Over 90% of SMA patients harbor a deletion of SMN(T), but relatively few base-pair mutations have been reported. We report here a novel G279C mutation with a G to T transversion on exon 7 (nucleotide position 868) of SMN(T). Another missense mutation has been reported recently on position 869. The fact that two mutations on the same codon both result in SMA suggest a functional significance of this amino acid within the SMN protein.

Amino Acid Substitution↗

Diet design for a multicenter controlled feeding trial: the DELTA program. Delta Research Group.

OBJECTIVE: To describe the process and results of diet standardization, diet validation, and monitoring of diet composition, which were key components of protocol 1 of Dietary Effects on Lipoproteins and Thrombogenic Activity (DELTA-1), the initial protocol in a program of multicenter human feeding studies designed to evaluate the effects of amount and type of fat on lipoproteins and hemostasis parameters in various demographic groups. DESIGN: DELTA-1 was based on a randomized, blinded, crossover experimental design. Three diets were fed for 8 weeks to 103 healthy men and women aged 22 to 67 years at 4 field centers. Diet A, an average American diet, was designed to provide 37% of energy from fat, 16% of energy from saturated fatty acids (SFAs); diet B (step 1 diet) was designed to provide 30% of energy from fat, 9% of energy from SFA; and diet C (low SFA diet) was designed to provide 26% of energy from fat, 5% of energy from SFA. Key features of diet standardization included central procurement of fat-containing foods, inclusion of standard ingredients, precision weighing of foods--especially sources of fat and cholesterol--and use of standardized written procedures. SETTING: For menu validation, a set of 12 menus for each diet was prepared in duplicate and chemically assayed. For monitoring of diet composition during the study, an 8-day diet cycle (6 weekday and 2 weekend menus) was sampled by every field center twice during each of 3 feeding periods. STATISTICAL ANALYSES: Means (+/- standard error) were calculated and compared with target nutrient specifications. RESULTS: DELTA-1 was able to provide a standardized diet that met nutrient specifications across 4 field centers over 24 weeks of participant feeding spanning a total of 8 months. APPLICATIONS: Prestudy chemical validation of menus and continuous sampling and assay of diets throughout the study are essential to standardize experimental diets and to ensure that nutrient target goals are met and maintained throughout a controlled multicenter feeding study.

Adult↗

Dobutamine stress echocardiography in the prediction of acute or chronic myocardial infarction.

BACKGROUND: We assessed the value of dobutamine stress echocardiography (DSE) in predicting cardiac events in patients with acute or chronic myocardial infarction (MI), and we studied the association between DSE and these events. METHODS AND RESULTS: Two hundred sixty-six patients (mean [+/-SD] age 65.3 +/- 11.4 years) with acute (n = 139) or chronic (n = 127) MI were recruited from March 1995 through April 1997. Both groups underwent DSE and were followed up for an average of 14.1 +/- 8.0 months. DSE was positive in 111 (79.9%) patients with acute MI and 65 (51.2%) patients with chronic MI (P <.0001 ). Positive DSE results were associated with a higher rate of all cardiac events (cardiac mortality rate, reinfarction, and unstable angina) than negative DSE results in both patients with acute MI and patients with chronic MI (44 in 111 patients vs 6 in 28 patients, P =.052, and 31 in 65 patients vs 10 in 62 patients, P <.0001, respectively). Among patients with acute MI, the positive and negative predictive values of DSE for all cardiac events were 39.6% (95% confidence interval [CI] 30.5% to 48. 7%) and 78.6% (95% CI 63.4% to 93.8%), respectively. In chronic MI, the positive and negative predictive values were 47.7% (95% CI 35.5% to 59.8%) and 83.9% (95% CI 74.7% to 93.0%), respectively. In both acute (P =.03) and chronic (P <.0001 ) MI, positive DSE findings were independent predictors of all cardiac events. CONCLUSIONS: DSE is useful for predicting cardiac events. A positive finding on DSE is an independent predictor of cardiac events after both acute and chronic MI, whereas a negative DSE result predicts a low likelihood of subsequent cardiac events.

Acute Disease↗

Effects of hormone replacement therapy on cardiovascular risk factors in postmenopausal women.

OBJECTIVE: To investigate changes in plasma lipoprotein profile, hemostatic factors, platelet aggregation, endothelin-1, and cardiac function during postmenopausal sequential 6-month hormone replacement therapy (HRT). DESIGN: Open longitudinal prospective study. SETTING: Gynecologic department of a medical center. PATIENT(S): Twenty-one healthy hysterectomized postmenopausal women. INTERVENTION(S): Oral E2 valerate (2 mg/d) combined with medroxyprogesterone acetate (MPA) (10 mg/d) during the last 10 days of each 21-day cycle. The treatment period was 6 months. MAIN OUTCOME MEASURE(S): Plasma lipoprotein profile, hemostatic parameters, platelet aggregation, endothelin-1, and left ventricular function. RESULT(S): After 6 months of treatment, total cholesterol, triglyceride, and low density lipoprotein (LDL) cholesterol were significantly progressively reduced. Atherogenic indices of total cholesterol-to-high-density lipoprotein (HDL) cholesterol and LDL-to-HDL cholesterols also showed a significant progressive decline. The concentrations of antithrombin III were significantly increased. The maximum aggregation and slope of platelet aggregation were significantly reduced, but all parameters were more pronounced at 1 month of HRT than at 3 or 6 months. The concentrations of endothelin-1 were significantly reduced (by 16.1%). In the evaluation of left ventricular function, only peak atrial diastolic velocity was significantly reduced. CONCLUSION(S): Combined HRT had favorable effects on lipids and lipoproteins, hemostatic factors, platelet aggregation, endothelin-1, and left ventricular function. However, further study is needed to evaluate the long-term effects of combined HRT, especially on platelet aggregation and cardiac function.

Blood Coagulation Factors↗

Chromosomal abnormalities of a new nasopharyngeal carcinoma cell line (NPC-BM1) derived from a bone marrow metastatic lesion.

An epithelial cell line, NPC-BM1, was established from a bone marrow biopsy of a female Taiwanese patient with nasopharyngeal carcinoma (NPC). Histopathology of the bone marrow biopsy and xenografts grown in severe combined immunodeficiency mice showed that the tumor was a nonkeratinizing, poorly differentiated carcinoma. NPC-BM1 cells grown as monolayers had a doubling time of 28.5 hours. Chromosome analysis showed that NPC-BM1 had the following features: 1) hypotetraploidy with a modal chromosome number of 87 (84-90); 2) numerically and structurally normal chromosomes 18; 3) numerical abnormalities without apparent structural alterations on chromosomes 14, 16, 17, 19, and 20; 4) ten structural abnormalities, t(1;9)(p11;q11), t(3;?;4)(p13;?;q13), add(4p),del(6p), i(8) [corrected] (q10),der(?)t(?;12),(?;p12),[corrected] add(21)(p11), del(X)(q24), add(X)(q22), and marker 1 (M1), in all metaphases examined, which were found to be present in two to five cell lines from primary NPC tumors reported previously; and 5) four other abnormalities, t(2;?;2)(p11.2;?;q21),t(11;22)(q11;q11),i(22)(q10), and marker 2 (M2), unique to this metastatic cell line. To the best of our knowledge, NPC-BM1 is the first NPC cell line derived from a distant metastatic site. Further evaluation of this cell line and additional metastatic NPC cell lines as well as primary NPC cell lines with respect to relations between the timing, karyotypic anomalies, and immunobiological characteristics in NPC progression and metastasis is warranted.

Adult↗