Objective To investigate the effect of resveratrol(Res) on neonatal rat cardiomyocyte lesion induced by hypoxia. Methods The cardiomyocytes of neonatal rats were cultured in vitro and the model of cardiomyocytes hypoxia was established. The cardiomyocyte vitalities were determined by MTT assay. The cardiomyocyte apoptosis was detected by Hoechst33258 fluorescent staining. The levels of total antioxidant capacity(T-AOC) and glutathione peroxidase(GSH-Px) were measured as well. Results After the administration of hypoxia for 8, 12, 16 and 24 hours, the cardiomyocyte inhibitory was(22.13?3.22)%, (29.75?0.34)%, (37.43?6.42)% and (45.47?7.32)%, respectively. After the administration of hypoxia for 24 hours, the typical morphologic changes of apoptosis in cardiomyocytes were showed.The intracellular GSH-Px activity decreased from(46.96?8.36)U/ml to(27.13?4.76)U/ml (P

In order to better fulfill the tasks of research,to turn out more quality papers,to produce outstanding results,and to further strengthen management and supervision of scientific research,the"quantitative economic management of scientific research quotas" was established in the hospital.Applying of the measure in scientific research management in the past eight years it was shown that the desired results were achieved,the academic advancement and the personnel growth were greatly promoted.

Objective To investigate function of Arg327Ile (R327I) and Arg327Ala(R327A) FⅨ mutants and to study the molecular pathogenesis of haemophilia B(HB) caused by R3271 mutation.Methods Hygromycin-resistant cell line was screened and the secretion of FⅨ antigen into the medium was measured by ELISA.The cell line with appropriate expression levels of F Ⅸ antigen was selected for culture.Recombinant F Ⅸ (rF Ⅸ ) was purified from concentrated medium by two step methods of Q-Sepharose Fast Flow and anion exchange chromatography.The concentration and purity of rF Ⅸ were determined by ELISA and SDS-PAGE,respectively.The activation of wild-type ( WT),R327I and R327A of rFⅨ by FⅦa/TF/Ca2+ or FⅪa/Ca2+ was identified by Western blot in different time periods.The FⅨa and FⅧa complex formed by interaction with different concentrations of FⅧa was used to activate F X,the apparent dissociation constant (Kd) for FⅧa binding was calculated by the kinetic results.The kinetic data of the activation of FX by WT,R327I and R327A FⅨa with or without FⅧa were calculated.Results The amount of WT,R327I and R327A rFⅨ were 450,210,64 μg,and the purity of rFⅨ was confirmed by SDSPAGE.Both R3271 and R327A could be normally activated by FⅧa/TF/Ca2+ or FⅪa/Ca2+.Kd for FⅧa binding showed that the binding capacities of R327I and R327A were 4 and 5 times lower than WT,respectively.The catalytic efficiencies of R327I and R327A F Ⅸ a for F X were 6 and 8 times lower with FⅧa,and 3 and 7.4 times lower without F Ⅷ a,respectively.Conclusions R327I and R327A rF Ⅸ mutants impair their binding to the FⅧa.The site on R327 contributes to FⅧa binding.It is partly related to the activation of FX.The low FⅧa binding to R327I FⅨa may cause HB.

Objective To discuss the tympanic membrane puncture or tympanostomy tube under endotoscope combined with adenoidectomy under nasal endoscopy on the influence of invalid conservative treatment of pediatric secretory otitis media.Methods A total of 112 cases of secretory otitis media with invalid conservative treatment were selected in the department of otorhinolaryngology of children′s hospital of Nanjing Medical University from July 2013 to August 2015.They were divided into three groups,the group A of 38 cases(68 ears)underwent transnasal endoscopic pure adenoidectomy;the group B of 37 cases(60 ears)underwent transnasal endoscopic adenoidectomy combined with tympanocentesis;the group C of 37 cases(59 ears)underwent tympanostomy tube insertion.The clinical treatment effect,the period of middle ear effusion,postoperative recurrence and complication were observed and compared.Results The total effective rate of the 3 groups was improved after 3 months(P<0.05);the total effective rate in group B and group C at one week and 3 months after operation were higher than those of group A(P<0.05);and the total effective rate in group C at 3 months after operation was significantly higher than that of group B(P<0.05).The level of auditory brainstem evoked potential(ABR)and the latency of ABR wave Ⅰ latency were decreased at 1 year after operation(P<0.05);the ABR changes in group B and group C at 1 week and 1 year after operation were lower than those in group A(P<0.05).The incidence of recurrence rate and the period of middle ear effusion in group C were lower than those in group A and B(P<0.05).The incidence of complication of group A was lower than those of group B and group C(P<0.05).Conclusion Using tympanostomy tube combined with adenoidectomy under endoscope can improve the children with hearing,which not only can shorten the time of the middle ear effusion,but also effectively reduce the recurrence rate.

Objective: To explore the effect of ISBAR communication mode on the bed shift in severe patients. Methods: The "ISBAR Nursing Delivery Card near the Bed of Severe Diseases Medicine" was formulated. 100 cases of severe patients admitted from February 2018 to March 2018 were set up as control group by experimental research methods. 100 cases of severe patients admitted from April 2018 to May 2018 were set up as experimental group and non-random control was established. The patients in the experimental group were treated with ISBAR nursing shift card for bed shift, and the control group were treated with normal oral shift. Observe and compare the incidence of nurse shift problems, the score of nurse shift assessment scale and the nurse′s knowledge of the patient′s condition "ten know" score between the two groups. Results: The incidence of nurse shift problems dropped from 39.50% (79/200) of control group to 16.50% (33/200) of experimental group, which had a significant difference (χ²=42.938, P<0.01). The score of the nurse′s succession assessment table was increased from (57.80 ± 3.61) points of control group to (73.96 ± 2.33) points of experimental group, which had a significant difference (t=-26.10, P<0.05). The nurse′s knowledge of the patient′s condition "ten know" had been increased from (72.14±4.13) points of control group to (87.75±2.54) points of experimental group, which had a significant difference (t=-32.01, P<0.05). Conclusion The use of ISBAR communication mode for serious patients in the severe medical department can effectively reduce the incidence of nurse shift problems, effectively improve nurses 'satisfaction with shift and the degree of mastery of patients′ conditions, and the effect of shift is better.

OBJECTIVE: To investigate incidents and clinical features of auditory neuropathy in Nanjing deaf school students. METHOD: Three hundred and fifty-eight deaf students in the school accepted the first examination including otoscopic examination, tympanometry and transiently evoked otoacoustic emissions (TEOAE) screening. Detailed audiological and vestibular evaluations including pure-tone audiometry, immittance audiometry and acoustic reflex measures, transiently evoked otoacoustic emissions (TEOAE), distortion product otoacoustic emissions (DPOAE), auditory brain stem response (ABR), electronystagmography (ENG) and vestibular evoked myogenic potential (VEMP) were given to whom had positive TEOAE screening. RESULT: Three hundred and twenty-three students entered the program of screening for auditory neuropathy. One student had positive TEOAE in single ear while the other two had positive TEOAE in both ears. In the screening stage,there were strong evidences in these three students with auditory neuropathy in the detailed audiological procedures. CONCLUSION Auditory neuropathy, which can also be found in deaf schools, is not as rare as we thought before. Early identification and intervention may help those children to avoid entering the deaf school and to return to normal society.
Adolescent ; Adult ; Audiometry, Pure-Tone ; Child ; China ; epidemiology ; Cochlear Microphonic Potentials ; Deafness ; epidemiology ; physiopathology ; Evoked Potentials, Auditory, Brain Stem ; Female ; Hearing Loss, Central ; epidemiology ; physiopathology ; Humans ; Male ; Otoacoustic Emissions, Spontaneous ; Students ; Young Adult

OBJECTIVETo investigate the effects of resveratrol (Res) on neonatal rat cardiomyocyte lesion induced by hypoxia.

METHODThe cardiomyocyte of neonatal rats were cultured in vitro and the model of cardiomyocyte hypoxia was established. The cardiomyocyte vitalities were determined by MTT assay, the HIF-1alpha expression levels in myocardial cells was detected by immunohistochemical, the activities of peroxidase (GSH-Px) and lactate dehydrogenase (LDH) were measured as well.

RESULTAfter the administration of hypoxia for 24 hours, the HIF-1alpha expression in myocardial cells was significantly increased. The LDH level in the culture medium was increased from (93.07 +/- 15.84) U x L(-1) to (750.77 +/- 181.51) U x L(-1) (P < 0.01). The intracellular GSH-Px activity was decreased from (46.96 +/- 8.36) U x mL(-1) to (27.13 +/- 4.76) U x mL(-1) (P < 0.05). Res 25, 50 and 75 micromol x L(-1) could dose-dependently inhibit the raising of the HIF-1alpha expression in myocardial cells induced by hypoxia. The LDH activities were decreased dose-dependently to (486.17 +/- 69.97), (189.43 +/- 32.07), (155.34 +/- 29.57) U x L(-1), respectively (P <0.05 or P <0.01). The GSH-Px activities were increased dose-dependently (33.55 +/- 6.34), (37.67 +/- 6.73), (41.44 +/- 7.91) U x mL(-1) (P < 0.05 or P < 0.01).

CONCLUSIONRes has a protective effect on neonatal rat cardiomyocyte lesion induced by hypoxia.

Animals ; Animals, Newborn ; Cell Hypoxia ; physiology ; Cells, Cultured ; Dose-Response Relationship, Drug ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Myocytes, Cardiac ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stilbenes ; pharmacology

Objective To identify the clinical features, the molecular diagnosis and the molecular mechanism of three unrelated factor X deficiency families. Methods Three probands were male and the diagnosis was validated by coagulant parameters. The F X coagulation activity ( F X∶ C ) and antigen (FX∶ Ag) were tested by clotting test and ELISA method. The cross-corrected test was used to rule out the inhibitor of FX in plasma. Thrombin generation test was evaluated. The antigen and the molecule weight of the FX in plasma were measured with western blotting. Gene mutations were analyzed in the probands and their family members with PCR and DNA sequencing. FX expression plasmids were constructed and transientby being transfected into 293T cells. FX: C and FX: Ag of the expression products were tested. Results APTT and PT in proband 1 were obviously prolonged, 113.4 s and 62.3 s, respectively. And there was no inhibitor in plasma. The thrombin generation was lower compared to normal reference. APTT and PT in proband 2 were 56. 5 s and 28.7 s. There was no inhibitor in the plasma. The thrombin generation was 1 101.5 nmol · min. APTT and PT in proband 3 were 117.3 s and 44. 3 s. The thrombin generation was 782.5 nmol · min. FX∶ C and FX∶ Ag in proband 1 were 1.4% and 3.6%, with a homozygous mutation in FX gene (Ser425→Pro). In vitro expression of the mutation showed a normal synthesis in the cell but secretion dysfuntion. In proband 2 F X: C and F X: Ag were 2. 2% and 5. 5%, with two heterozygous mutations in FX gene (Ala-29→Pro and Phe324→Leu). The Ala-29 → Pro mutation led to significantly reduced expressions of FX in both cell lysate and cell culture supernatants compared to wild-type plasmid,(41.32 ±5.21 )% and(6. 30 ± 1.84)% respectively. However Phe324→Leu mutation almost did not affect the FX synthesis. FX: C and FX: Ag in proband 3 were 2. 2% and 35%, with two heterozygous mutations in FX gene( Ala235→Thr and Arg347→Cys). The expressions of these two mutant FX proteins in cell lysate were similar to those of wild-type but obviously lower in the supernatant. Conclusions Five mutations of F X gene are found in this study. These mutations (Ser425Pro, Phe324Leu, Ala235Thr and Arg347Cys)can not affect F X protein synthesis. However Ala-29Pro mutation can reduce F X protein synthesis and cause secretion dysfunction.

OBJECTIVETo study the molecular pathogenesis of protein C (PC) deficiency in a patient with pulmonary embolism and in his family members.

METHODSAnticoagulated blood samples were collected from the proband and his family members to detect PC, PS and AT activities. PC antigen level was measured using ELISA. The genomic DNA was extracted to amplify all the 9 exons and their flanking sequences of PC gene using PCR, and the PCR products were sequenced. The mutated exons identified were amplified and sequenced for the other family members.

RESULTSThe proband and his parents and sister were identified as carriers of PC gene mutation, which led to type II PC deficiency. Sequencing of the proband's PC gene showed two heterozygous point mutations in exon 3 (G5540A) and exon 7 (C10230T) to cause compound heterozygous mutations of PC E29K and PC R147W, which were inherited from his father and mother, respectively. His sister was a heterozygote of PC R147W.

CONCLUSIONThe proband is a compourd heterozygous mutations carrier of PC E29K and PC147W. PC E29K is a novel PC mutation, and PC R147W is a reported PC gene mutation seen in patients with type II hereditary PC deficiency and recurrent thrombosis.

Adolescent ; Base Sequence ; Heterozygote ; Humans ; Male ; Molecular Sequence Data ; Pedigree ; Point Mutation ; Protein C ; genetics ; Protein C Deficiency ; complications ; genetics ; pathology ; Pulmonary Embolism ; etiology ; genetics

Objective: To investigate the clinical features and surgical treatment effect of patients with cardiac cavernous hemangioma. Methods: Clinical data of 9 patients (5 male, aged from 4 to 53 years old) with cardiac cavernous hemangioma, who underwent surgical treatment from November 2002 to March 2015 and the diagnosis of cardiac cavernous hemangioma was confirmed by postoperative histological examination, were retrospectively analyzed. Effects of surgical treatment were analyzed. Results: Four patients were asymptomatic (heart murmur presented in 3 patients during physical examination). Three patients presented with palpitation, chest distress, and short of breath. One patient presented with epigastric discomfort and another patient presented with intermittent fever for more than 10 months. ST and T wave changes of electrocardiogram were found in 2 patients, cardiac mass was detected in the right heart chamber in 5 patients by echocardiography, and no cardiac mass was detected the rest 4 patients. Cardiac masses were resected en bloc, then the adjacent tissues were repaired in 7 patients, and mass was partially resected due to the involvement with adjacent heart structure. No cardiac mass was found during operation in 1 case, impaired mitral valve structure was excised and postoperative pathologically confirmed as cardiac valve cavernous angioma on the excised mitral valve structure. No signs of recurrence or enlargement of cardiac cavernous hemangioma were found during the 11(10, 11)years follow up. Conclusions There is no specific clinical feature for patients with cardiac cavernous hemangioma. It is difficult to detect the cardiac valve cavernous angioma by echocardiography before surgery. Individualized surgical treatment is associated with good clinical outcome in this patient cohort. However, the clinical features and surgical treatment effect of patients with cardiac cavernous hemangioma still need to investigate in large sample trial.