OBJECTIVETo explore the effect of Salvianolate on myosin heavy chain (MHC) in cardiomyocytes of congestive heart failure (CHF) rats.

METHODSSixty male SD rats were divided into 6 groups according to random digit table, i.e., the normal control group (NCG), the model group, the Captopril group (CAG), the low dose Salvianolate group (LSG), the high dose Salvianolate group (HSG), the Captopril and high dose Salvianolate group (CSG), 10 in each group. CHF rat model was established with peritoneal injection of adriamycin in all rats except those in the NCG. Equal volume of normal saline was peritoneally injected to rats in the NCG, once per week for 6 successive weeks. Corresponding medication was started from the 5th week of injecting adriamycin. Rats in the CAG were administered with Captopril solution at the daily dose of 10 mg/kg by gastrogavage. Rats in the LSG and the HSG were administered with Salvianolate solution at the daily dose of 24.219 mg/kg and 48.438 mg/kg respectively by gastrogavage. Salvianolate was dissolved in 2 mL 5% glucose solution and administered by peritoneal injection. Rats in the CSG were peritoneally injected with high dose Salvianolate solution and administered with Captopril solution by gastrogavage. Two mL normal saline was peritoneally injected to rats in the model group, once per day for 8 successive weeks. Eight weeks later, the cardiac function and myocardial hypertrophy indices were detected by biological signal collecting and processing system. mRNA expression levels of alpha-MHC and beta-MHC in cardiac muscle were detected by fluorescence quantitative PCR. Expressions of protein kinase C (PKC) in cardiac muscle were detected by Western blot.

RESULTSCompared with the normal control group, heart mass index (HMI) and left ventricular mass index (LVMI) obviously increased in the model group (P < 0.01). Compared with the model group, HMI and LVMI decreased in HSG, CAG, and CSG groups (P < 0.05, P < 0.01). It was more obviously lowered in the CSG group than in the CAG group (P < 0.05). Compared with the NCG, the mRNA expression level of alpha-MHC in cardiac muscle decreased, the mRNA expression level of p-MHC and the expression of PKC in cardiac muscle increased in the model group (P < 0.01). Compared with the model group, the mRNA expression level of alpha-MHC in cardiac muscle was increased, and the mRNA expression level of beta-MHC and the expression of PKC in cardiac muscle were decreased in HSG, CAG, and CSG groups (P < 0.05, P < 0.01). There was statistical difference between the CSG group and the CAG group (P < 0.05).

CONCLUSIONSSalvianolate could up-regulate the mRNA expression level of alpha-MHC, and down-regulate the mRNA expression level of beta-MHC in cardiac muscle. Its mechanism might be related to decreasing the expression of PKC.

Animals ; Captopril ; Doxorubicin ; Drugs, Chinese Herbal ; Heart Failure ; metabolism ; Male ; Myocardium ; Myocytes, Cardiac ; drug effects ; metabolism ; Myosin Heavy Chains ; metabolism ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley

Objective To analyze the karyotypes of 11 cases of Turner syndrome with marker chromosome,and study the phenotypic effects resulting from the abnormal karyotype.Methods Eleven Turner syndrome patients had a mosaic karyotype and carried a marker chromosome,and 6 marker chromosomes were ring chromosomes.Their karyotypes were showed as mos.45,X/46,X,+mar or mos. 45,X/46,X,+r.Fluorescence in situ hybridization(FISH)technique with X/Y centromere probes was performed to determine the origin of the marker chromosome.Reverse chromosome painting technique was used to identify the breakpoints of two largest markers.Phenotype effects with different chromosome breakpoints were compared.Results All the 11 marker chromosomes were ring X chromosomes.The breakpoints of the r(X)were involved in Xp22,Xq22,Xq24 and Xq26,etc.Conclusions The marker chromosomes in Turner syndrome mainly originate from X chromosome and form ring chromosome X.Each r (X)in our patients was mosaic,indicating it was originated from mitosis error during early embryo development.To analyze the origin of the marker chromosome and the breakpoint of r(X)will provide guidance for the therapy and prognosis of the Turner syndrome patient.

Objective To evaluate the influence of gestational diabetes mellitus (GDM) on maternal and perinatal outcomes in twin pregnancies. Methods We retrospectively analyzed the clinical features of both twin and singleton pregnancies, which delivered in Guangzhou Women and Children's Medical Center between January 1, 2012 and December 31, 2013. The twin pregnancies were divided into two groups:those with (GDM-T, n=51) and without GDM (non-GDM-T, n=130), which were matched by maternal age and delivery time (within one month) in a ratio of 1∶2 among singleton pregnancies with (GDM-S, n=102) and without GDM (non-GDM-S, n=102), respectively. The differences of adverse maternal and perinatal outcomes among these four groups were examined. The overall assessment of pregnancy outcomes was completed using Delphi method. Statistical analysis was performed with one-way analysis of variance, t test, Kruskal-Wallis test, rank test, Chi-square test or Fisher's exact test. Results (1) When compared to GDM-S and non-GDM-S group respectively, less women conceived with the help of assisted reproductive technology, higher proportion of women underwent and gestational age at delivery tend to be earlier in GDM-T and non-GDM-T group (all P<0.01). In oral glucose tolerance test,the fasting blood glucose level of GDM-T group was higher than the other three groups (F=21.716, P<0.01), the glucose levels at 1 and 2 h were higher than non-GDM-T and non-GDM-s respectively (both P<0.01), but no significant difference was found when compared with GDM-S group (P>0.01). Similarly, no significant difference was found in prenatal glycosylated hemoglobin value between GDM-T and GDM-S group (P>0.01). (2) There was no significant difference in the incidences of hypertensive disorders of pregnancy, anemia, premature rupture of membranes, oligohydramnios, placental abruption, postpartum hemorrhage, asphyxia neonatorum, small for gestational age, hypoglycemia of newborn, hyperbilirubinemia of newborn and perinatal death between GDM-T group and the other three groups(all P>0.01). Higher incidences of hypertensive disorders of pregnancy and postpartum hemorrhage were shown in the GDM-T group than in the GDM-S and non-GDM-S groups, respectively (both P<0.01). The incidences of preterm birth in GDM-T and non-GDM-T group were both higher than that in GDM-S and non-GDM-S, respectively [54.9%(66/102), 53.8%(140/260), 5.0%(10/102) and 3.0%(6/102), all P<0.01], while no significant difference was found between GDM-T and non-GDM-T group (P>0.01). (3) The overall assessment of pregnancy outcomes did not show any difference between GDM-T group and the other three groups (χ2=6.707, P>0.01). However, the score for fetal outcomes in the GDM-T group was higher than in the GDM-S and non-GDM-S group, but lower than in non-GDM-T group [M(Q)=1.0(2.3), 0.0(3.0), 0.0(0.0), 1.0(2.8) score, χ2=122.818, P<0.01]. Conclusions GDM does not increase the risk of adverse pregnant outcomes in twin pregnancies.

Objective To explore the role of white matter injuries in the schizophrenia induced by the NMDA re-ceptor antagonist. Methods Adult male C57BL/6J mice (8 week old) were equally divided into four groups. One group was sub-chronically treated with saline solution, and the other three groups were intraperitoneally treated with MK-801 at dose of 0.025 mg/mL (M1), 0.050 mg/mL (M2) and 0.100 mg/mL (M3) in a volume 10 ml per kilogram body weight. All ani-mals were tested using Morris water maze at the 9th-15th day and using the Hole Board exploration as well as Rota Rod performance tests on the 16th day. The myelin basic protein (MBP) and the ultrastructure of the myelin sheaths in the cor-pus callosum were then examined using immunohistochemical methods, transmission electron microscope technique and stereological methods. Results The repeated sub-chronic MK-801 treatment did not induce impairment of spatial learning and memory in Morris water maze. The MK-801 treatment at 0.25 mg/kg and 1.00 mg/kg but not at 0.50 mg/kg resulted in less exploration to a new environment. The myelin staining with anti-MBP antibody was less intense in all three schizo-phrenic groups when compared to saline control group (P<0.01). Furthermore, MK-801 treatment caused pathological al-terations of the myelin sheaths including segmental demyelination of myelinated fibers and splitting of myelin sheath lamel- lae in schizophrenic groups. The ratio of the injured myelinated nerve fibers in the corpus callosum of MK-801 treated mice [M3 group, (22.42 ± 4.24)%] was significantly higher when compared to the control mice [(3.84 ± 1.35)%,P<0.01)]. Conclusions The present study demonstrated the white matter damages, mainly low MBP expression and segmental demye-lization in the corpus callosum in the mice sub-chronic treated with MK-801, indicating that the white matter changes might be involved in the schizophrenia induced by NMDA antagonist.

OBJECTIVEOtoacoustic emissions (OAE) and auditory brainstem responses (ABR) are tests widely used in neonatal hearing screening. This study aimed to investigate the differences and clinical value of distortion product otoacoustic emissions (DPOAE) and ABR in hearing screening of high-risk neonates admitted to a neonatal intensive care unit (NICU).

METHODSDPOAE and ABR were measured with the Smart-OAE analyser and the Smart-EP brain-stem electric response audiometry apparatus, respectively, in 600 high-risk neonates (1,200 ears). The testing results of DPOAE and ABR were compared.

RESULTSOf the 600 neonates (1,200 ears), the incidence of ABR abnormality (78.6%, 943/1,200) was remarkably higher than that of DPOAE abnormality (22.3%, 268/1,200). Two hundred and forty-one ears (20.8%) were negative and 252 (21%) were positive in both DPOAE and ABR tests. A total of 707 ears (58.9%) presented with a discordant result in DPOAE and ABR. The false positive and false negative rates of the DPOAE test were 6.0% (16/268) and 74.1% (691/932) respectively.

CONCLUSIONSIn high-risk neonates the diagnostic value of DPOAE for identification of hearing loss, when used alone, is limited. The ABR test appears to be more reliable for hearing screening in high-risk neonates. It is suggested that hearing screening for high-risk neonates should be conducted with ABR first, followed by OAE after failure on ABR.

Evoked Potentials, Auditory, Brain Stem ; physiology ; Female ; Hearing Tests ; methods ; Humans ; Infant, Newborn ; Intensive Care, Neonatal ; Male ; Neonatal Screening ; methods ; Otoacoustic Emissions, Spontaneous ; physiology

OBJECTIVETo study the relation of cytochrome C release from mitochondria to cytosol and neuronal apoptosis after cerebral hypoxia-ischemia (HI) in neonatal rats.

METHODSHypoxia-ischemia was induced in 7-day-old rat pups by ligation of left carotid artery and 7.7% oxygen was inhaled for 55 min. The pups were sacrificed and the brains were taken out at different recovery time. Some of the brains were homogenized and cellular fraction of mitochondria and cytosol was isolated with different speed centrifugation. The cellular fraction was used for Western blotting. Some of the brains were sectioned and stained with antibody against cytochrome C and TUNEL as well as double labeling with different combinations.

RESULTSWestern blots showed that cytochrome C in mitochondria was not reduced significantly at 1 h, but reduced markedly at 14 h in ipsilateral hemisphere post-HI. However, the immunoreactivity of cytochrome C in cytosol was increased markedly at 1 h post-HI and reached peak at 14 h post-HI. The number of cytochrome C positive cells in the cortex was increased significantly at 1 h (8.4 +/- 1.8/visual field) compared to normal control (1.5 +/- 0.8/visual field) (P < 0.01) and reached peak at 14 h (29.0 +/- 5.2/visual field) post-HI. The number of TUNEL positive cells increased significantly at 1 h post-HI (14 +/- 3/visual field) compared to normal control (1.5 +/- 0.8/visual field) (P < 0.01) and reached peak at 24 h (286 +/- 86/visual field). The double labeling of cytochrome C and active caspase-3 showed that they colocalized well at 3 h after HI. Furthermore, the positive cells showed nuclei condensation. There were more active caspase-3 positive cells at late recovery (24 h and on) after HI. The double labeling of cytochrome C and TUNEL showed only part of Positive cells colocalized. The cells with cytochrome C strong staining showed TUNEL negative or weakly positive. The cells with TUNEL strong staining showed weakly cytochrome C staining.

CONCLUSIONCytochrome C release is one of the early biochemical changes of neuronal apoptosis after hypoxia-ischemia in neonatal rat brain.

Animals ; Animals, Newborn ; Apoptosis ; Blotting, Western ; Caspase 3 ; Caspases ; metabolism ; Cytochromes c ; analysis ; secretion ; Female ; Hypoxia-Ischemia, Brain ; metabolism ; pathology ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Mitochondria ; metabolism ; Rats ; Rats, Wistar ; Time Factors

Objective To investigate the added diagnostic value of SPECT/CT imaging over routine planar whole-body bone scintigraphy (WBBS) for bone metastases from lung cancer. Methods One hundred and forty-six patients with lung cancer, confirmed by pathological examination, underwent routine 99Tcm-MDP (1110 MBq) WBBS, followed by SPECT/CT over the regions with indeterminate findings on WBBS. Both WBBS and bone SPECT/CT images were interpreted by two experienced nuclear medicine physicians in consensus as the positive, negative or uncertain bone metastases. The final diagnosis was comfirmed by pathology or clinical follow-up. x2 test was used to compare the differences between the two imaging methods. Results Finally, 45 patients were diagnosed as positive bone metastases and the other 101 as negative. The diagnostic sensitivity of bone SPECT/CT for bone metastases from lung cancer was 93.3%(42/45), singnificantly higher than that of WBBS (64.4%, 29/45) (x2 = 19.944, P＜0.05). The diagnostic accuracy of bone SPECT/CT was 89.7% ( 131/146), much higher than that of WBBS (44.5%,65/146) ( x2 = 69. 598,P ＜ 0.05). The uncertain and incorrect diagnostic rates of bone SPECT/CT and WBBS were 10.3% ( 15/146, raging from 5.3% to 15.2% with 95% confidence interval (CI) ) and 55.5% (81/146, raging from 47. 4% to 63.5% with95% CI), respectively. Conclusion BoneSPECT/CT provides incremental diagnostic value over routine WBBS for bone metastases from lung cancer.

Objective To investigate the additional diagnostic value of 99Tcm-MDP SPECT/CT over conventional SPECT scan in patients with spinal bone malignancy. Methods Fifty-two patients (mean age (56±14) years) with suspicious spinal bone diseases underwent both bone SPECT and SPECT/CT imaging right after 99Tcm-MDP whole-body planar bone scintigraphy. All patients were pathologically diagnosed by either spine operation or trans-spinal biopsy. The images were evaluated by two independent reviewers; inter-reviewer agreement was evaluated using a weighted Kappa score. Each focus of abnormality was recorded using a 4-point diagnostic confidence scale: benign (B), possibly benign (PB), possibly malignant (PM) or malignant (M). Results Accord to the pathological results, 36 patients had bone malignancy (19 with metastatic tumors and 17 with non-metastatic tumors) and 16 had benign lesions. Over SPECT images, two reviewers rated 73.1% (38/52) and 67.3% (35/52) the lesions as the equivocal (PB or PM) respectively. Over SPECT/CT images, they rated only 25.0% (13/52) and 13.5% (7/52) lesions as the equivocal. Inter-reviewer agreement was 63.5% (weighted Kappa score=0.62) for SPECT and 78.9% (weighted Kappa score=0.81) for SPECT/CT. Conclusions Compared with99Tcm-MDP SPECT, 99Tcm-MDP SPECT/CT results in a significant reduction of equivocal diagnoses for the spinal bone malignancy.

Objetive: To investigate whether apoptosis of SGC7901 cells can be induced by the expression of the recombinant gene of anti-HER2 ScFv/tBid. Methods: The recombinant anti-HER2 ScFv/tBid gene was cloned into vector pCMV and the recombinant plasmid was transfected into SGC7901 cells. The gene expression was detected by RT-PCR and immunofluorescent staining. Cell counting was carried out to show the effect of the gene transfection on cell growth. At the same time, significant apoptotic peak was detected by flow cytometry in recombinant anti-HER2 ScFv/tBid gene transfected cells. Results: The fusion protein of anti-HER2 ScFv/tBid was observed in the cytoplasm of transfected SGC7901 cells. The transfected cells displayed typical cell growth inhibition and apoptosis. Conclusion: Fusion protein of anti-HER2 ScFv/tBid can induce apoptosis of SGC7901.

OBJECTIVETo determine the karyotype of a boy suspected to have Cri du Chat syndrome with severe clinical manifestations, and to assess the recurrence risk for his family.

METHODSHigh-resolution GTG banding was performed to analyze the patient and his parents. Fluorescence in situ hybridization (FISH) with Cri du Chat syndrome region probe as well as subregional probes mapped to 5pter, 5qter, 18pter, 18qter, and whole chromosome painting probe 18 was performed to analyze the patient and his parents. In addition, single nucleotide polymorphism-based arrays (SNP-Array) analysis with Affymetrix GeneChip Genome-wide Human SNP Nsp/Sty 6.0 were also performed to analyze the patient.

RESULTSKaryotype analysis indicated that the patient has carried a terminal deletion in 5p. FISH with Cri du Chat syndrome region probe confirmed that D5S23 and D5S721 loci are deleted. SNP-Array has detected a 15 Mb deletion at 5p and a 2 Mb duplication at 18p. FISH with 5p subtelomeric probes and 18p subtelomeric probe further confirmed that the derivative chromosome 5 has derived from a translocation between 5p and 18p, which has given rise to a 46,XY,der(5)t(5;18)(p15.1;p11.31)dn karyotype.

CONCLUSIONA de novo 5p partial deletion in conjunction with a cryptic 18p duplication has been detected in a boy featuring Cri-du-Chat syndrome. His parents, both with negative findings, have a low recurrence risk. For its ability to detect chromosomal imbalance, SNP-Array has a great value for counseling of similar patients and assessment of recurrence risks.

Child, Preschool ; Chromosome Banding ; Chromosome Deletion ; Chromosomes, Human, Pair 18 ; Chromosomes, Human, Pair 5 ; Cri-du-Chat Syndrome ; diagnosis ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Phenotype ; Polymorphism, Single Nucleotide ; Trisomy