BACKGROUND: Epoxyeicosatrienoic acids (EETs), which are metabolites of arachidonic acid catalyzed by cytochrome P450 epoxygenase, are degraded into inactive dihydroxyeicosatrienoic acids by soluble epoxide hydrolase (sEH). Many studies have revealed that sEH gene deletion exerts protective effects against diabetes. Vascular calcification is a common complication of diabetes, but the potential effects of sEH on diabetic vascular calcification are still unknown. METHODS: The level of aortic calcification in wild-type and Ephx2-/- C57BL/6 diabetic mice induced with streptozotocin was evaluated by measuring the aortic calcium content through alizarin red staining, immunohistochemistry staining, and immunofluorescence staining. Mouse vascular smooth muscle cell lines (MOVAS cells) treated with β-glycerol phosphate (0.01 mol/L) plus advanced glycation end products (50 mg/L) were used to investigate the effects of sEH inhibitors or sEH knockdown and EETs on the calcification of vascular smooth muscle cells, which was detected by Western blotting, alizarin red staining, and Von Kossa staining. RESULTS: sEH gene deletion significantly inhibited diabetic vascular calcification by increasing levels of EETs in the aortas of mice. EETs (especially 11,12-EET and 14,15-EET) efficiently prevented the osteogenic transdifferentiation of MOVAS cells by decreasing nidogen-2 (NID2) expression. Interestingly, suppressing sEH activity by small interfering ribonucleic acid or specific inhibitors did not block osteogenic transdifferentiation of MOVAS cells induced by β-glycerol phosphate and advanced glycation end products. NID2 overexpression significantly abolished the inhibitory effect of sEH gene deletion on diabetic vascular calcification. Moreover, NID2 overexpression mediated by adeno-associated virus 9 vectors markedly increased insulin-like growth factor 2 (IGF2) and phospho-ERK1/2 expression in MOVAS cells. Overall, sEH gene knockout inhibited diabetic vascular calcification by decreasing aortic NID2 expression and, then, inactivating the downstream IGF2-ERK1/2 signaling pathway. CONCLUSIONS sEH gene deletion markedly inhibited diabetic vascular calcification through repressed osteogenic transdifferentiation of vascular smooth muscle cells mediated by increased aortic EET levels, which was associated with decreased NID2 expression and inactivation of the downstream IGF2-ERK1/2 signaling pathway.
Animals ; Mice ; Vascular Calcification/metabolism* ; Mice, Inbred C57BL ; Epoxide Hydrolases/metabolism* ; Diabetes Mellitus, Experimental/genetics* ; Male ; Gene Deletion ; MAP Kinase Signaling System/genetics* ; Cell Line ; Immunohistochemistry ; Muscle, Smooth, Vascular/metabolism* ; Signal Transduction/genetics* ; Mice, Knockout

Objective:To explore the correlation between color Doppler ultrasound, Cyfra21-1, Fer and malignant ovarian lesions and the value of combined model identification.Methods:A total of 108 patients with suspected ovarian cancer admitted to our hospital from Jan. 2021 to Dec. 2023 were selected as subjects. All patients were tested by color Doppler ultrasound, Cyfra21-1 and Fer, and were divided into malignant group (61 cases) and benign group (47 cases) according to the pathological test results. The baseline data, ultrasound parameters, Cyfra21-1 and Fer levels of the two groups were compared, and multivariate regression analysis was performed to establish a multi-factor combined model, and ROC curve was used to evaluate the differential efficacy of the combined model.Results:There was no significant difference in baseline data such as age and course of disease between the two groups ( P>0.05). In the malignant group, the lesion diameter was (7.66±1.24) cm, with solid echo detection in 37 cases and inner wall with nipple detection in 38 cases. In the benign group, the lesion diameter was (5.14±1.03) cm, with solid echo detection in 18 cases and inner wall with nipple detection in 17 cases. In the malignant group, PS was (22.94±4.61) cm/s, EDV was (15.67±3.42) cm/s, VM was (12.43±3.15) cm/s. PS, EDV and VM in benign group were (15.63±4.24) cm/s, (8.95±3.04) cm/s and (17.08±4.21) cm/s respectively, and those in malignant group were higher than those in benign group ( P<0.05). RI in the malignant group was 0.35±0.06, RI in the benign group was 0.58±0.13, and RI in the malignant group was lower than that in the benign group ( P<0.05). Multivariate regression analysis showed that lesion diameter, solid echo, inner wall with papilla, PS, EDV, VM, Cyfra21-1, Fer and RI were all independent risk factors for ovarian malignant lesions, and lesion diameter, solid echo, inner wall with papilla, PS, EDV, VM, Cyfra21-1 and Fer were positively correlated with ovarian malignant lesions. RI was negatively correlated with ovarian malignant lesions ( P<0.05). ROC curve results showed that the diagnostic efficiency of the combined model was higher than that of the single use of Doppler ultrasound, Cyfra21-1 and Fer. Conclusion:The combined diagnosis model of color Doppler ultrasound, Cyfra21-1 and Fer is helpful to improve the diagnostic efficiency of ovarian malignant lesions.

A retrospective analysis of the clinical data of seven acute B-lymphoblastic leukemia (B-ALL) patients with TCF3-HLF fusion gene-positive admitted to the First Affiliated Hospital of Soochow University from June 2017 to August 2022 was conducted to summarize their clinical features and prognoses. The seven B-ALL patients comprised four males and three females, with a median age of 18 (11-33) years. Five patients tested positive for CD33 expression, and four patients had a normal karyotype. Two patients had hypercalcemia at the initial diagnosis, and one patient developed hypercalcemia at relapse. Six patients presented with coagulation dysfunction at diagnosis. After induction chemotherapy, five out of seven patients achieved complete remission, of which four subsequently relapsed. Two patients did not achieve remission even after two rounds of induction chemotherapy, with one achieving complete remission after treatment with blinatumomab immunotherapy. Three patients underwent chimeric antigen receptor T cell therapy, whereas three patients subsequently underwent hematopoietic stem cell transplantation. Five patients died, while two patients survived with sustained complete remission. TCF3-HLF-positive B-ALL is rare and has a high relapse rate and poor prognosis.

Objective To explore the status-quo and possible constraints of scientific research in a municipal public hospital,provide countermeasures for the improvement of scientific research administration capacity.Methods Questionnaire survey was conducted to 1 356 medical staff in a tertiary hospital in Zhejiang province,information collected including the statusquo of scientific research,attitude towards scientific research,difficulties and suggestions.Results Among 1 316 valid questionnaires,61 ％ of the respondents agreed that scientific research was very important for the development of hospitals,83 ％ of the respondents were willing to use their spare time to conduct scientific research projects,and 61.2％ of them usually had plans to do research but did not know where to start.The main constraints identified were the poor academic atmosphere,lack of scientific research training,and lack of scientific research facilities and resources.Conclusions The medical staffs in a municipal public hospital have high subjective enthusiasm for scientific research,low personal research ability and poor research environment.It is suggested to improve the scientific research ability of the staff on the basis of improving the objective environmental conditions for research.

Objective To study thO application valuO of ultrasound guidOd-corO nOOdlO biopsy(US-CNB) in thO diagnosis of brOast lOsions.Methods From January 2016 to April 2018,51 patiOnts with brOast massOs who undOrwOnt US-CNB in thO POoplO′s Hospital of TongchOng wOrO sOlOctOd. ThO pathological rOsults aftOr surgOry wOrO usOd as thO gold standard, and thO puncturO succOss ratO, sOnsitivity, spOcificity and accuracy of US -CNB wOrO analyzOd.Results Forty-ninO lOsions wOrO diagnosOd by qualitativO diagnosis, of which 42 lOsions wOrO malignant and 7 lOsions wOrO bOnign. Two lOsions wOrO diagnosOd by qualitativO diagnosis, and thO succOss ratO of puncturO was 96.1% (49/51).ThO postopOrativO pathology confirmOd that 44 casOs wOrO malignant and 6 casOs wOrO bOnign. ThO sOnsitivity, spOcificity and accuracy of US-CNB wOrO 97.7% (42/43),100.0% (42/42),98.0% (48/49),rOspOc-tivOly.Conclusion ThO succOss ratO and accuracy of US-CNB puncturO in thO diagnosis of bOnign and malignant lOsions arO high, consistOnt with thO gold standard. It has clinical significancO for thO basic hospital lOvOl.

Objective To investigate the anti-inflammatory and analgesic effects of extracts from Garden Balsam seeds in order to provide theoretical basis for rational clinical application. Methods The mouse ear swelling induced by xylene, the body torsion induced by acetic acid or hot plate method, the rat foot swelling induced by egg white and granuloma model were set up to study anti-inflammatory and analgesic effects of extracts from Garden Balsam seeds. Results The water extracts at the middle and high doses reduced the body torsion counts of mice (P<0. 05), alleviated the rat toe swelling, but did not affect mice ear swelling. The extracts at the high dose prolonged pain threshold at 60 min (P<0. 01) and 90 min (P<0. 05); the alcohol ex-tracts at the middle and high doses significantly lowered body twisting counts (P<0. 01), attenuated the ear swelling of mice (P<0. 05), remarkably prolonged the pain threshold (P<0. 01)at 30 min, weakened the rat foot swelling, and elevated 60 min pain threshold at the high doses (P<0. 01), but the extracts exerted no effects on the rats granuloma. Conclusion The extracts from Garden Balsam seeds have anti-inflammatory and analgesic effects.

Aim To research dynamically the changes of endogenous cystathionine- β-synthase/hydrogen sul-fide system in PC12 cells injury induced by rotenone. Methods Rotenone-induced injury in PC12 cells ( characteristic of dopaminergic neurons) was used as a PD cell model. The expression of CBS was evaluated by Western blot. Intracellular CBS activity and H2 S production were detected by Methylene blue spectro-phot-ometric method. The viability of PC12 cells was measured by CCK-8 assay. GSH detection kit was used to detect the intracellular GSH content. Results In the groups of 6 and 12 hours, the expression and activ-ity of CBS were elevated, and the production of H2 S was increased. In the groups of 24 and 48 hours, CBS expression and activity were significantly decreased, and the amount of H2 S was significantly reduced. Ap-plication of 1. 5 μmol·L-1 rotenone for different time (6-48h) could decrease the cell viability and intra-cellular GSH contents in a time-dependent manner. Conclusions The expression and activity of endoge-nous CBS, stimulated by rotenone, are elevated firstly and then decreased. The generation of H2 S, stimulated by rotenone, is increased and then reduced significant-ly, which may be related to PC12 cells against oxida-tive stress damage induced by rotenone.

OBJECTIVETo explore the clinical and laboratory features of chronic myeloid leukemia (CML) with atypical e14a3 and e19a2 BCR-ABL fusion gene subtypes.

METHODSWe retrospectively analyzed a cohort of CML patients with Ph chromosome positive confirmed by cytogenetic and FISH but classical e13a3(b2a2), e14a2(b3a2)and e1a2 fusion transcripts negative identified by conventional real-time quantification RT-PCR (RQ-PCR). Further RQ-PCR was done with the forward primer and reverse primer designed to detect rare atypical BCR-ABL fusion genes including e14a3 and e19a2 transcripts. Direct sequencing analysis was performed on the PCR products and mutations in the BCR-ABL kinase domain were detected. The clinical data of patients were retrospectively analyzed.

RESULTSSix CML patients were found to carry t(9;22) abnormality and BCR-ABL rearrangement confirmed by FISH but classical BCR-ABL fusion genes negative detected by RQ-PCR. Further RQ-PCR and sequencing analysis confirmed the fusion of BCR exon 14 and ABL exon 3 in five CML patients (case 1-5) and the fusion of BCR exon 19 and ABL exon 2 in one CML patient (case 6). E255K and I293T IM-resistant mutations were detected in case 1 and 2, respectively. Among five cases with e14a3 transcripts, four were CML-CP, one CML-AP. Four patients were male and one was female. The median age was 48 years. The patient (case 6) with e19a2 transcripts was 40-year-old female with a diagnosis of CML-CP and PLT count was more than 1 000×10⁹/L. Imatinib (IM) therapy was administer in case 1, 2, 3, 4 and hematopoietic stem cell transplantation (HSCT) was undergone in case 5 after hydroxyurea (Hu) or interferon failure. Case 1 who had E255K IM resistant mutation, responded poorly to IM but obtained a complete cytogenetic remission (CCyR) after a substitution of dasatinib for IM. Case 2 and 3 achieved CCyR 6 months later after IM treatment and had been maintained well with IM despite I293T mutation in case 2. Case 4 attained CCyR 3 months later after IM treatment but relapsed and died soon. Case 5 was still in CCyR after HSCT. Case 6 with e19a2 transcripts got complete hematologic response after Hu treatment and CCyR was achieved soon after IM therapy.

CONCLUSIONIncidence of CML with atypical transcripts is extremely low. They could benefit from tyrosine kinase inhibitors or HSCT. Rare and atypical BCR- ABL fusion gene subtypes could be missed by conventional RQ-PCR.

Adult ; Female ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; classification ; diagnosis ; genetics ; Male ; Middle Aged ; Retrospective Studies

Objective To investigate the level of follicular fluid NO, VEGF and ET-1 in assessing IVF-ET outcome. Methods Totally 131 patients undergoing IVF-ET cycles were recruited. The level of follicular fluid NO was measured by chromatometry. The follicular fluid VEGF and ET-1 were measured by ELISA. Transvaginal ultrasound was performed on human chorionic gonadotropin (hCG) injection day to determine ovarian volume and antral follicle count. Results The pregnancy rate was 37.40% (49/131). There were significantly increased level of follicular fluid NO, VEGF and decreased level of follicular fluid ET-1 in the pregnant group than those in the non-pregnant group (P<0.05). Total ovarian volume and antral follicle count on HCG injection day were significantly higher in the pregnant group than those in the non-pregnant group (P<0.05). The levels of follicular fluid NO and VEGF had positive correlations with the total ovarian volume and antral follicle count. However, the level of follicular fluid ET-1 had a negative correlation with the total ovarian volume and antral follicle count. Conclusions The high level of NO, VEGF and low level of ET-1 in follicular fluid are good predictors of ovarian blood flow and ovarian response in IVF-ET.

OBJECTIVE:To study the pharmacokinetics of boanmycin and to make a clinical evaluation on which.METHODS:36patients with malignant tumor were subjected to the first stage of clinical study after injected intramuscular injection.with0.5～7.5mg/m 2 boanmycin.Of which,8patients were chosen as the subjects for the pharmacokinetic study by microbioassay.RESULTS:Boanmycin had little influence on the functions of heart,liver,kidney and lungs and which had no inhibitory action on bone marrow.After intramuscular injection with5.29～7.65mg/m 2 boanmycin,parameters of pharma?cokinetics in the patients were determined as the following:t 1/2? was(14.036?4.409)min,t 1/2? was(39.160?5.599)min,AUC was(14.697?1.826)(?g?min)/ml and CLs was(0.781?0.102)L/min.CONCLUSION:Serum clearance of boanmycin shows two-compartment model.