Study on pathological mechanism and establishment of animal model of ICU acquired weakness / 医学研究生学报

ObjectiveThere are few animal experiments on ICU acquired weakness (ICU AW), and suitable animal models are the main constraints. The study was to explore the method of ICU AW animal model which satisfies the clinical requirements of ICU and suitable for large-scale animal trials.MethodsThirty six SD rats were randomly divided into Control group(0.9%NS at 2.5 ml/kg intraperitoneal injection once a day for consecutive 3 days), immobilization group(the left hindlimb was immobilizated for 7d, then the immobilization was removed to 14 d), sepsis group(lipopolysaccharide at 2.5 mg/kg intraperitoneal injection once a day for consecutive 3 days) and sepsis-immobilization group(the left hindlimb was immobilizated for 7d, then the immobilization was removed to 14 d and lipopolysaccharide at 2.5 mg/kg intraperitoneal injection once a day for consecutive 3 days).To determine whether the model was successful, the muscle strength of left hindlimb, gastrocnemius/body weight ratio and pathological changes of gastrocnemius were measured at 0 d( immediately after intervention), 3d, 7 d, 10 d, 14 d. To explore the possible pathological mechanism, creatinine/body weight ratio, albumin, lymphocyte, and gastrocnemius pathological scores were measured.Results7 days later, the scores of left hindlimb muscle strength and pathology in sepsis immobilization group were significantly higher than those in the sepsis group and the control group(P<0.05), contary body weight and gastrocnemius weight were lower than those in control group, immobilization group and sepsis group(P<0.05), and gastrocnmiu/body weight ratio(0.528±0.018) was significantly lower than those in control group(0.756±0.315) and sepsis group(0.813±0.040)(P<0.05). Creatinine / body weight in sepsis immobilization group(0.283±0.0268) was significantly higher than those of blank group (0.185±0.022), immobilization group (0.207±0.027) and sepsis group (0.246±0.043)(P<0.05). The lymphocyte count [(5.193±1.493) ×109/L] was significantly lower than that in the blank group[(7.005±0.702) ×109/L] and the immobilization group[(7.208±0.832) ×109/L)](P< 0.05). 14 days later, the scores of left hindlimb muscle strength, body and gastrocnemius weight in sepsis immobilization group were significantly lower than those in control group, immobilization group and sepsis group(P<0.05). Gastrocnmiu/body weight ratio in sepsis immobilization group(0.519± 1.493) was significantly lower than those in control group(0.798±0.015), immobilization group (0.570±0.022)and sepsis group(0.693±0.022)(P<0.05).ConclusionThe qualified animal model of ICU AW can be established by repeated intraperitoneal injection in low dose of lipopolysaccharide combined with limb immobilization. Immunosuppression and Hypercatabolism in ICU AW rats is an important reason that ICU AW can not to be mitigated. Thus, we supposed that it may be the mechanisms for the development of ICU AW,which needs further experimental verification.

Investigation on the Medical Reference Range of the Normal Neonates with the Amino Acid and Acyl Carnitine Tandem Mass Spectrometry in Gansu Province / 现代检验医学杂志

Objective To establish the medical reference range of amino acid and acyl car-nitine tandem mass spectrometry in normal neonates in Gansu province,and provide the basis for the determination of amino acid and acyl carnitine test results in the screening of neonatal genetic metabolic diseases.Methods A non derivatization tandem mass spectro-metry kit was used to screen 77 957 samples of neonates in Gansu province.Statistical soft ware SPSS19.0 was used to analyze the 95％ reference range of 11 amino acids and 31 kinds of acylcarnitine indexes.Results The medical reference value(μmol/L) of the series mass spectra of newborn genetic metabolic diseases in Gansu province was established:ALA (216.17 ～ 727.58),ARG (1.80～33.03),CIT (4.87～30.67),GLY (183.43～841.46),LEU (79.85～289.45),MET (3.32～25.86),ORN (34.09～225.15),PHE (27.04～83.37),PRO (79.44～337.59),TYR (37.61～177.79),VAL (59.31～250.95),C0 (9.35～45.35),C2 (2.62～25.40),C3 (0.46～3.3),C3DC_C4OH (0.02～0.20),C4 (0.08～0.31),C4DC_C5OH (0.10～0.32),C5 (0.05～0.30),C5:1 (0.00～0.01),C5DC_C6OH (0.04～0.22),C6 (0.01～0.06),C6DC (0.03～0.13),C8 (0.02～0.08),C8:1 (0.05～0.31),C10 (0.02～0.12),C10:1 (0.03～0.10),C10:2 (0.01～0.10),C12 (0.02～0.13),C12:1 (0.02～0.10),C14 (0.07～0.30),C14:1 (0.03～0.13),C14:2 (0.01～0.03),C14OH (0.00～0.02),C16 (0.59～4.91),C16:1 (0.03～0.30),C16:1-OH (0.01～0.08),C16OH (0.01～0.03),C18 (0.29～1.30),C18:1 (0.57～2.32),C18:1-OH (0.01～0.05),C18:2 (0.08～0.51) and C18OH (0.00～0.02).Conclusion The establishment of the medical reference range of the normal neonatal dryblood spot amino acid and acyl carnitine tandem mass spectrometry in Gansu province can provide reference for the determination of the results of the cluster mass spectrometry in this region.

Preparation and antitumor effects of tanshinone ⅡA loaded albumin nanoparticles / 中国中药杂志

In this study, the tanshinone ⅡA loaded albumin nanoparticles were prepared by high pressure homogenization method. The formulation was optimized by central composite design-response surface method (CCD-RSM), with the particle size, encapsulation efficiency, and drug loading as indexes to investigate their in vitro anti-tumor effect. The results showed that the prepared nanoparticles had uniformly spherical morphology and uniform particle size distribution. The average particle size, encapsulation efficiency and drug loading of nanoparticles were about (175.7± 3.07) nm, 90.8%±1.47% and 5.52%±0.09%, respectively. Tanshinone ⅡA loaded albumin nanoparticles showed a more powerful antitumor effect than free tanshinone ⅡA for human promyelocytic leukemia NB4 cells. The preparation method of the drug-loaded albumin nanoparticles was simple and easy, and can significantly improve the solubility of tanshinone ⅡA, so it was helpful to extend its application in therapies against hematological malignancies.

Preparation of CD123 mono-antibody modified tanshinone ⅡA loaded immunoliposome and its in vitro evaluation / 中国中药杂志

In the study, we developed a novel formulation, CD123 mono-antibody (mAb) modified tanshinone ⅡA loaded immunoliposome (CD123-TanⅡA-ILP) to achieve the targeted drug delivery for leukemia cells. Orthogonal test was used to optimize liposome preparation, and the TanⅡA-loaded PEGylated liposomes (TanⅡA-LP) of S100PC-Chol-(mPEG2000-DSPE)-TanⅡA at 19∶5∶1∶1 molar ratio were prepared by the thin film hydration-probe ultrasonic method. A post-insertion method was applied to prepare CD123-TanⅡA-ILP via thiolated mAb conjugated to the terminal of maleimide-PEG2000-DSPE. The cellular uptake assay was measured by flow cytometry, and the inhibitory effect of CD123-TanⅡA-ILP on NB4 cells proliferation was tested by using MTT assay. The results of cellular uptake assay showed that CD123-ILP could significantly increase the drug uptake of NB4 cells as compared with free drugs and LP. The IC₅₀ values at 48 h incubation were 20.87, 11.71, 7.17 μmol•L⁻¹ respectively for TanⅡA，TanⅡA-LP and CD123-TanⅡA-ILP. CD123-ILP demonstrated a potential and promising targeted drug delivery strategy for acute myelogenous leukemia (AML) treatment.

Effect of licorice flavonoids on kainic acid-induced seizure in mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of licorice flavonoid (LF) on kainic acid (KA)-induced seizure in mice and its mechanism.</p><p><b>METHODS</b>Male adult ICR mice were injected with 25 mg/kg KA to induce temporal lobe seizure. LF was administrated 7 d before seizure induction (pre-treatment) or 24 h after seizure induction (post-treatment) for 7 d. Acute seizure latency, seizure stage and duration were observed and compared between LF- and vehicle-treated mice. From d2 on, mice with status epilepticus were video-monitored for spontaneous seizures, 10 h/d for 6 w. Immunohistochemical analysis of BrdU and Timm staining was conducted to detect the neurogenesis and mossy fiber sprouting, respectively.</p><p><b>RESULTS</b>No significant difference was observed in acute seizure latency, seizure stage and duration between LF-and vehicle-treated mice. KA-induced acute seizure resulted in spontaneous seizure in mice, and the seizure frequency was increased with time. Pre- and post-treatment with LF decreased seizure frequency from w3 after modeling [(0.58±0.15)/d, (0.38±0.38)/d vs (1.23±0.23)/d, P <0.05]. Furthermore, KA-induced seizure resulted in robust neurogenesis and mossy fiber sprouting, while treatment with LF both pre- and post- KA injection significantly inhibited neurogenesis (15.6±2.6, 17.1±3.1 vs 28.9±3.5, P <0.05) and mossy fiber sprouting (1.33±0.31, 1.56±0.42 vs 3.0±0.37, P <0.05).</p><p><b>CONCLUSION</b>LF has no significant anti-seizure effect. However, it can decrease epileptogenesis through inhibition of neurogenesis and mossy fiber sprouting.</p>

Effect of IGF-1 on long-term anxiety-like behavior in rats after hypoxic-ischemic brain damage / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To observe the changes in anxiety-like behavior among rats in the recovery stage after hypoxic-ischemic brain damage (HIBD) during the perinatal period and to investigate the effect of insulin-like growth factor 1 (IGF-1) on the long-term anxiety-like behavior and its action mechanism among rats with HIBD.</p><p><b>METHODS</b>Ninety neonatal rats (7 days old) were randomly and equally divided into normal control, HIBD, and HIBD+IGF-1 groups. A neonatal rat model of HIBD was established by Rice method in the HIBD and HIBD+IGF-1 groups. The rats in the HIBD+IGF-1 group were intraperitoneally injected with IGF-1 (0.2 mg/kg) immediately after HIBD, and the other two groups were intraperitoneally injected with an equal volume of normal saline. The anxiety-like behavior was evaluated by elevated plus-maze test on postnatal days 21 and 28. The expression of tyrosine hydroxylase (TH) in the substantia nigra was measured by immunohistochemistry on postnatal days 14, 21, and 28.</p><p><b>RESULTS</b>On postnatal days 21 and 28, the open-arm time (OAT) and percentage of OAT for the HIBD and HIBD+IGF-1 groups were significantly lower than those for the normal control group (P<0.05), but there were no significant differences between the HIBD and HIBD+IGF-1 groups (P>0.05); the percentage of open arm entry showed no significant difference between the three groups (P>0.05). On postnatal day 14, there were no significant differences in percentage of TH immunostaining-positive area between the three groups (P>0.05). On postnatal days 21 and 28, the HIBD and HIBD+IGF-1 groups had significantly lower percentages of TH immunostaining-positive area than the normal control group (P<0.05), but there was no significant difference between the HIBD and HIBD+IGF-1 groups (P>0.05).</p><p><b>CONCLUSIONS</b>HIBD in the perinatal period may cause the changes in anxiety-like behavior in adolescent rats, which may be related to decreased expression of TH in the substantia nigra. Neonatally given IGF-1 cannot improve the long-term anxiety-like behavior in rats after HIBD, and it does not affect TH expression in the substantia nigra. IGF-1 may not regulate the changes in long-term anxiety-like behavior in adolescent rats.</p>

Clinical features and treatment of acute clenbuterol poisoning in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study clinical features, treatment and curative effects in children with acute clenbuterol poisoning, in order to provide a basis for early diagnosis and treatment.</p><p><b>METHODS</b>Clinical data of 28 hospitalized children with acute clenbuterol poisoning in April 2011 were retrospectively studied.</p><p><b>RESULTS</b>Of the 28 patients, there were 15 males and 13 females, aged 1 to 13 years (mean age 6.5±4.8 years). Vomiting, palpitations and limb shaking were found as main clinical manifestations in the patients. Main changes of blood biochemical included hypokalemia, lactic acidosis, hyperglycemia, hypsocreatinkinase. Snus tachycardia and S-T segment depression were observed on ECG. Patients' symptoms were gradually alleviated after 12-78 hours by use of beta blockers, potassium supplement, protecting the heart and other symptomatic and supportive treatment. Blood biochemical indexes were improved after 48 hours of admission. All of the patients were cured after 5 days. The symptoms of the patients do not longer occur during a follow up of half a month.</p><p><b>CONCLUSIONS</b>Acute clenbuterol poisoning is characterized by vomiting, palpitations, limb shaking, hypokalemia, lactic acidosis and tachycardia in children. An early effective treatment of this disease can improve prognosis in children.</p>

Changes in Th17 and CD4+CD25+ Treg cells and their significance among children with hand, foot and mouth disease / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the changes in peripheral blood Th17 and CD4(+)CD25(+) regulatory T (Treg) cells and their significance among children with hand, foot and mouth disease (HFMD).</p><p><b>METHODS</b>Eighty-nine children with HFMD, including 55 cases of common HFMD and 34 cases of severe HFMD, were included in the study; and 30 healthy children were selected as the control group. The percentages of Th17 and CD4(+)CD25(+) Treg cells in CD4(+) T cells in peripheral blood were determined by flow cytometry. The expression levels of interleukin (IL)-10, transforming growth factor-β (TGF-β), and IL-17 were measured by enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>Compared with the control group, the cases of common HFMD and severe HFMD had significantly increased levels of Th17 cells and IL-17 (P<0.05) but significantly decreased levels of CD4(+)CD25(+) Treg cells, IL-10, and TGF-β (P<0.05). The severity of the HFMD was positively correlated with the levels of Th17 cells and IL-17 in peripheral blood but negatively correlated with the levels of CD4(+)CD25(+) Treg cells, IL-10, and TGF-β.</p><p><b>CONCLUSIONS</b>Children with HFMD have increased response of Th17 cells but decreased response of CD4(+)CD25(+) Treg cells in peripheral blood. Th17/CD4(+)CD25(+) Treg cell imbalance may play an important role in the pathogenesis of HFMD.</p>

Effect of rhIGF-1 on cytochrome C and caspase-3 expression in neonatal rats with hypoxic-ischemic brain damage in vivo / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effect of rhIGF-1on the mRNA and protein expression of cytochrome C (Cyt-C) and caspase-3 in neonatal rats with hypoxic-ischemic brain damage (HIBD).</p><p><b>METHODS</b>Ninety neonatal Sprague-Dawley rats were randomly divided into three groups: normal control, HIBD, and HIBD+rhIGF-1 (rhIGF-1 was given intraperitoneally right after HI). Rat HIBD model was prepared according the Rice-Vannucci method. RT-PCR and Western blot methods were used to measure the mRNA and protein expression of Cyt-C and caspase-3 24, 48 and 72 hrs after HI (n=10 each time point).</p><p><b>RESULTS</b>At all time points, both Cyt-C mRNA and caspase-3 mRNA expression levels in the HIBD group increased compared with those in the normal control group, and those in the HIBD+rhIGF-1 group also increased compared with that in the normal control group but decreased compared with that in the HIBD group. There were statistical significances among the three groups (P<0.01). At all time points, the changes of both Cyt-C and caspase-3 protein expression in the three groups were similar to those of the mRNA expression: both Cyt-C and caspase-3 protein expression levels increased in the HIBD group compared with those in the normal control group, and those in the HIBD+rhIGF-1 group also increased compared with those in the normal control group but decreased compared with those in the HIBD group. There were statistical significances among the three groups (P<0.01).Pearson correlation analysis showed that mRNA and protein expression of Cyt-C were positively correlated to casapse-3 mRNA and protein expression in the HIBD and the HIBD+rhIGF-1 groups.</p><p><b>CONCLUSIONS</b>rhIGF-1 may inhibit the Cyt-C release and caspase-3 expression, and thus provides neuroprotection against HIBD in neonatal rats.</p>

Pathologic changes and expression of Heme oxygenase-1 in paraquat-induced renal injury / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the mechanism of paraquat-induced renal injury in rats.</p><p><b>METHODS</b>Adult healthy Sprague-Dawley (SD) rats (female and male in half) were randomly divided into two groups, the control group and the paraquat poisoned group. The rats in the paraquat poisoned group were treated with PQ (25 mg/kg) intraperitoneally while the rats in the control group were treated with the same dose of normal saline. Its histopathological change was observed and the expression of HO-1 and the mRNA expression of HO-1 were detected by RT-PCR at 3rd h, 6th h, 12th h, on 1st d, 2nd d, 3rd d and 5th d.</p><p><b>RESULTS</b>(1) In the control group, the tissue structure was clear without edema, vacuolar degeneration, cloudy swelling and necrosis. In the paraquat poisoned group, there were obvious lesions in the renal tubule of cortical part, including cellular swelling, the narrow cannula, the mesenchymal congestion and edema. These pathologic changes gradually became more severe, reached the peak on the 1st day, and did not relieve until the end of this study; there was the karyopyknosis and the cyto-architecture disappeared in some severe cases; Some glomerulus and medulla were also involved. (2) In the control group, there was no or weak expression of HO-1 and HO-1 mRNA. At the 3rd hour, the expressions of HO-1 in the paraquat poisoned group were observed in the membrane and cytoplasm of renal tubular epithelial cell of cortical part. Immunohistochemistry score (IHS) in the paraquat poisoned group was higher than that in the control group (P<0.05), except the HIS of the 5th day. At the 3rd hour, the expression of HO-1 mRNA increased, reached the peak on the 1st day, and then decreased. The expression of HO-1 mRNA was (0.53 +/- 0.21), (0.55 +/- 0.31), (0.56 +/- 0.22), (0.64 +/- 0.14) and (0.43 +/- 0.25) at the time point other than on the 3rd and 5th day. It showed statistical difference between the paraquat poisoned group and the control group from the 3rd hour to the 2nd day (P<0.05).</p><p><b>CONCLUSION</b>The mechanism of paraquat induced-renal injury is multiple. The higher expression of HO-1 and HO-1 mRNA were involved in the procedures of paraquat-induced renal injury.</p>

Study on the relationship between somatic health and incidence of depression in the elderly / 中华流行病学杂志

<p><b>OBJECTIVE</b>In order to explore the risk factors of geriatric depression, a longitudinal follow-up study was conducted on elderly population living in the community so as to provide evidence for the development of depression prevention and control.</p><p><b>METHODS</b>A sampled population consisting 2506 elderly was selected from urban and rural communities in Beijing, using well-established sampling techniques as cluster, stratification and random selection. Data was collected by trained staff members, using standard survey instruments in 2000 and 2004.</p><p><b>RESULTS</b>Longitudinal study showed that the four-year cumulative incidence of the geriatric depression in Beijing was 10.58%. Difference on were evident intelligence/education, with the rates for illiteracy (15.2%) and primary school (10.5%) significantly higher than that of junior high school and above (5.1%) (chi2 = 26.587, P = 0.000). Rates also varied substantially with place of residence, individuals living in rural areas had a substantially higher rate of depression (15.4%) than those individuals dwelling in urban district (6.1%) (chi2 = 31.163, P = 0.000). Poor self-rated health condition (chi2 = 23.385, P = 0.000), cognitive impairment (chi2 = 11.947, P = 0.001) and limitations in physical functioning (ADL: chi2 = 15.930, P = 0.000; IADL: chi2 = 9.501, P = 0.002) were related to the worsening of depressive symptoms. Results from logistic regression analysis indicated that education level, dwelling area, self-rated health condition and ADL were the independent risk factors.</p><p><b>CONCLUSION</b>Lower educational level, dwelling situation, poor self-rated health condition as well as ADL damage might increase the incidence of depression, suggesting more attention needs to be paid to improve somatic function of elderly in order to decrease the incidence of geriatric depression and to improve the prognosis of the disease and the quality of life.</p>

Hypermethylation of promoter region of RASSF1A gene in ovarian malignant epithelial tumors / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate hypermethylation of promoter region of RASSF1A and its relationship with ovarian malignant epithelial tumors.</p><p><b>METHODS</b>Methylation-specific PCR (MSP) was used to determine the hypermethylation of promoter region of ras association domain family 1 (RASSF1A) gene in 80 cases of ovarian malignant epithelial tumors.</p><p><b>RESULTS</b>No methylation of promoter region of RASSF1A gene was found in all 80 normal control tissues (0). Of 80 ovarian malignant epithelial tumors 42 were hypermethylated in promoter region of RASSF1A gene (52.5%). There was no statistically significant difference in the frequency of hypermethylation of RASSF1A gene among serious adenocarcinomas, mucinous adenocarcinomas and endometrioid adenocarcinomas (54.2%, 52.4% and 45.5%, respectively; P > 0.05). Hypermethylation of RASSF1A gene happened more often in tumors in stage III and IV (66.7% and 77.8%) than that in stage I and II (21.4% and 16.7%; P < 0.05). It was less frequently observed in well and moderately differentiated tumors (34.5% and 35.0%) than in poorly differentiated tumors (80.6%; P < 0.05).</p><p><b>CONCLUSION</b>High frequency of methylation of RASSF1A promoter exists in ovarian malignant epithelial tumors as a tumor suppressor gene, its suppressor activity may be abrogated by an epigenetic mechanism. Hypermethylation of RASSF1A promoter in patients with epithelial malignant ovarian tumors is related to clinical stage and histopathological grade. It indicates poor prognosis.</p>

Expression of RASSF1A and RASSF1C transcripts in human primary ovarian cancers / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the expression of two major alternative transcripts of RASSF1 gene (RASSF1A and RASSF1C) in human primary ovarian cancers and their biological implication as a new tumor suppressor gene.</p><p><b>METHODS</b>Reverse transcription-polymerase chain reaction (RT-PCR) and laser capture microdissection (LCM) were used to determine mRNA expression of the two major alternative transcripts of RASSF1 gene (RASSF1A and RASSF1C) in 3 ovarian cancer cell lines and 80 cases of primary ovarian cancers.</p><p><b>RESULTS</b>RASSF1A mRNA was undetectable in SK-OV-3 cell line. Expression of RASSF1A and RASSF1C in 80 primary ovarian cancers were 40.0% (32/80) and 91.3% (73/80) respectively. RASSF1A mRNA expression was detectable more frequently in stage I and II (71.4%, 10/14; 75.0%, 9/12) than in stage III and IV ovarian cancers (26.7%, 12/45; 14.1%, 1/9) (P < 0.05). The expression level was also higher in well and moderately differentiated tumor groups (58.6%, 17/29; 50.0%, 10/20) than in poorly differentiated tumor group (16.1%, 5/31) (P < 0.05).</p><p><b>CONCLUSION</b>There is a preferential loss of RASSF1A expression in human ovarian cancers and its expression is correlated with the tumor stage and the degree of histological differentiation which, as a tumor suppressor gene, might play an important role in the tumorigenesis of human primary ovarian cancer.</p>

Detection of circulating hypermethylated tumor-specific RASSF1A DNA in ovarian cancer patients / 中华病理学杂志

<p><b>OBJECTIVE</b>To detect hypermethylated tumor-specific RASSF1A DNA in the circulation and its significance in ovarian cancers patients.</p><p><b>METHODS</b>Methylation-specific polymerase chain reaction (MSP) was used to study the hypermethylation of RASSF1A in preoperative serum samples from 51 ovarian cancer patients.</p><p><b>RESULTS</b>The RASSF1A gene was not methylated in peripheral blood samples from 51 normal patients and 51 patients with benign ovarian tumors. Hypermethylation of RASSF1A gene was found in circulating tumor-specific DNA in 43.1% of patients (22 out of 51 cases) with ovarian cancers (P < 0.05). There was no difference in hypermethylation of RASSF1A gene amongst various ovarian cancer subtypes (P < 0.05). On the other hand, hypermethylation of RASSF1A gene was more frequently encountered in stage III and IV than stage I and II tumors (P < 0.05). It was rarely seen in well and moderately differentiated groups, as compared with poorly differentiated group (P < 0.05).</p><p><b>CONCLUSIONS</b>There is a higher frequency of RASSF1A hypermethylation in circulating tumor-specific DNA of ovarian cancer patients. RASSF1A has been postulated to play an important role as tumor suppressor gene and can be silenced by promoter hypermethylation. This methylation correlates with clinical stage and histopathologic grade. Such observation may carry diagnostic and prognostic implications when assessing ovarian tumors.</p>