Objective To construct the fibroblast-specific non-viral vector pcDNA3-CEP-BMP-2 containing collagen 1A2 enhancer and promoter,and to validate the enhancement of BMP-2 expression in the human dermal fibroblasts by this vector,compared with the routine non-viral BMP2 vector. Methods The sequences for collagen 1A2 enhancer and promotor,and BMP-2 gene were ligated into the pcDNA3 plasmids.The plasmids were transfected into human skin fibroblasts and vein endothelial cells by means of cationic liposomes.The expressions of the plasmids in these two kinds of cells were detected by RT-PCR.The osteogenic phonotypes of fibroblasts were determined.(Results)pcDNA3-CEP-BMP-2,which contained collagen 1A2 enhancer and promoter could enhance the BMP-2 expression in the fibroblasts but not in vein endothelial cells.Osteogenetic phenotypes were more obvious in the fibroblasts transfected with pcDNA3-CEP-BMP-2 than in pcDNA3-BMP-2-transfected ones. Conclusion Collagen 1A2 enhancer and promoter can enhance BMP2 expression in fibroblasts.

Objective: To analyze the prevalence and predictor for left ventricular reverse remodeling (LVRR) in patients of primary hypertension combining left ventricular systolic dysfunction (LVSD) with tailored medication.
Methods: A total of 118 consecutive patients admitted in our unit from 2010-08 to 2012-10 with the base line left ventricular ejection fraction (LVEF)≤40%were enrolled. The demographic and clinical information with the findings of echocardiography at admission were collected. The patients were followed-up until 2013-12 or until the all cause death/cardiac transplantation. According to echocardiography, LVRR was deifned by 2 criteria at the same time:①the absolute elevation of
LVEF≥10%than base line and the follow-up LVEF≥50%,②the relative reduction of left ventricular end-diastolic diameter (LVEDD) index≥10%than base line and the follow-up LVEDD index≤27 mm/m2. LVRR prevalence with its base line predictor was investigated.
Results: The overall mean follow-up time was (23 ± 15) months, and 39/118 (33.1%) patients acquired LVRR as LVEF from the base line level (30.6 ± 6.8)%increased to the follow-up level (57.0 ± 4.9)%;LVEDD index from the base line level (31.6 ± 3.9) mm/m2 decreased to the follow-up level (24.4 ± 1.9) mm/m2, all P<0.01. The average time length for reaching LVRR was (11 ± 9) months, and 27/39 (69.2%) patients reached LVRR within 12 months. There were 79 patients not reached to LVRR, while their LVEF also from the base line level (28.6 ± 6.1)%increased to the follow-up level (39.0 ± 13.2)%;LVEDD index from the base line level (38.1 ± 5.6) mm/m2 decreased to the follow-up level (36.1 ± 6.9) mm/m2. Multivariable logistic regression analysis indicated that the patients with the shorter duration of heart failure (HF) as>6 months vs≤6 months (OR=0.244, P<0.01), shorter QRS interval as≥120ms vs<120ms (OR=0.276, P<0.05) and the higher quartile of systolic blood pressure (SBP)/LVEDD index (OR=2.724, P<0.01) at admission were the independent predictors for LVRR.
Conclusion:With tailored medication, about 1/3 of patients with hypertension combining LVSD could acquire LVRR, the patients with shorter duration of HF, shorter QRS interval and higher ratio of SBP/LVEDD index had more possibilities.

Objective: To investigate the prevalence of hyponatremia and the relationship between hyponatremia and prognosis of dilated cardiomyopathy (DCM) for in-hospital patients. Methods: A total of 515 consecutive in-hospital DCM patients treated in HF center of Fu Wai Hospifal from 2008-10 to 2013-10 were retrospectively studied. Hyponatremia was deifned as the serum level of sodium < 135 mmol/L at ifrst admission. The prevalence of hyponatremia and the relationship between hyponatremia and DCM prognosis were studied including the risk of in-hospital time and mortality, the rates of all cause death and HF worsening death after discharge. Surviving patients were followed-up by clinical or telephone visit until 2014-11 or until the death. Results: There were 134/515 (26.0%) patients suffered from hyponatremia at admission, the serum level of sodium was related to HF symptom duration, NYHA functional classiifcation, systolic blood pressure (SBP), left atrial diameter and total bilirubin level, allP<0.01. Compared with non-hyponatremia, the patients with hyponatremia presented longer in-hospital time(14.8±11.1) days vs (11.2±5.8) days and higher in-hospital mortality (18.7% vs 1.8%), bothP< 0.01. There were 483 survivors discharged and were followed-up for (30.7 ± 19.5) months, during that period, the rates of all cause death and HF worsening death were 26.5% and 21.9% respectively. The patients with hyponatremia had the higher rates of all cause death (47.7% vs 20.3%) and HF worsening death (44.0% vs 15.5%), bothP<0.01. Multiple Cox regression analysis showed that with adjusted HF history (> 6 months vs≤ months ), NYHA functional classiifcation (Ⅱ-Ⅳ), SBP (per 10 mmHg elevation), total bilirubin level (per 1 mg/dl change) and LVEDD (per 5 mm change), the hyponatremia at admission is still one of the important independent predictors for all cause death (HR=1.836, 95% CI (1.248-2.702),P<0.01 and HF worsening death HR=2.139, 95% CI (1.406-3.253),P<0.01 in DCM patients after discharge. Conclusion: Hyponatremia is a common electrolyte disorder for in-hospital DCM patients, it is related to longer in-hospital time and higher mortality; higher rates of all cause death and HF worsening death after discharge in DCM patients.

Adult ; Biopsy, Needle ; Diagnosis, Differential ; Humans ; Kidney ; pathology ; Kidney Diseases ; pathology ; therapy ; Male ; Nephritis, Interstitial ; pathology ; Renal Dialysis ; Sarcoidosis ; pathology ; therapy ; Tuberculosis, Renal ; pathology

Objective:To observe the effect of herbal cake-partitioned moxibustion on the expressions of mitogen-activated protein kinase (MEK1/2) and extracellular regulatory protein kinase (ERK1/2) in gastric tissues of rats with spleen deficiency syndrome, and to explore the possible mechanisms of herbal cake-partitioned moxibustion in treating spleen deficiency syndrome. Methods:Sixty Sprague-Dawley (SD) rats were randomly divided into a blank control group (group A), a model group (group B), a ranitidine group (group C), and a herbal cake-partitioned moxibustion group (group D) by random digit, 15 rats in each group. Rat models of spleen deficiency syndrome were made by intragastric administration of 4℃ 200% concentrated Da Huang (Radix et Rhizoma Rhei). After successful modeling, the rats in group C were treated with 25 mg/(kg·bw) ranitidine by intragastric adminstration and rats in group D were treated with herbal cake-partitioned moxibustion at Zusanli (ST 36) and Zhongwan (CV 12), for 8 d. Excepted for rats in group A, all the other rats were treated with indomethacin at 5 mg/(kg·bw) at 8:00 a.m. on the second day after finishing all the intervention and sacrificed 7 h later to isolate the stomach. Histopathological changes of the gastric tissues were observed under light microscope after hematoxylin-eosin (HE) staining. The protein expressions of MEK1/2 and ERK1/2 in the gastric tissues were detected by immunohistochemistry. Results:After intervention, the gastric mucosal injury in group B was significantly severer than that in group A, with large breakage and ablating; the damage of gastric mucosa was decreased in group C compared with group B; the gastric mucosal surface remained relatively complete, and the status of breakage and ablating was significantly improved. After intervention, compared with group A, the protein expressions of MEK1/2 and ERK1/2 in gastric tissues of the other groups were significantly higher (P<0.01). Compared with group B, the protein expressions of MEK1/2 and ERK1/2 in group C and D were significantly higher (allP<0.01). Compared with group C, the protein expressions of MEK1/2 and ERK1/2 in group D were significantly higher (P<0.01). Conclusion: Herbal cake-partitioned moxibustion promotes the repair of gastric mucosa in rats with spleen deficiency syndrome, via improving protein expressions of MEK1/2 and ERK1/2 in gastric tissues, as well as activating MEK/ERK signaling pathway.

BACKGROUNDRecent autopsy study showed a high incidence of cerebrovascular lesions in Alzheimer's disease (AD). To assess the impact of cerebrovascular pathology in AD, we used diffusion tensor imaging (DTI) to study AD patients with and without cerebrovascular lesions.

MATERIALS AND METHODSConventional and DTI scans were obtained from 10 patients with probable AD, 10 AD/V patients (probable AD with cerebrovascular lesions) and ten normal controls. Mean diffusivity (D) and fractional anisotropy (FA) values of some structures involved with AD pathology were measured.

RESULTSD value was higher in AD patients than in controls in hippocampus and the cingulate gyrus. In AD/V patients, increased D value was found in the same structures and also in the thalamus and basal ganglia compared to controls. There was a significant difference of D value between AD and AD/V patients. FA value reduced in the white matter of left inferior temporal gyrus and in the bilateral middle cingulate gyrus in patients with AD/V compared with controls. The MMSE (mini-mental state examination) score significantly correlated with FA value in the right hippocampus (r=0.639, P<0.019), in the right anterior cingulate gyrus (r=0.587, P<0.035) and in left parahippocampal gyrus (r=0.559, P<0.047).

CONCLUSIONCerebrovascular pathology had stronger impact on the D value than the AD pathology alone did. Elevated D value in thalamic and basal ganglia may contribute to cognitive decline in AD/V patients. Reduced FA values in AD/V patients may indicate that cerebrovascular pathology induced more severe white matter damage than the AD pathology alone did.

Aged ; Alzheimer Disease ; complications ; pathology ; Brain ; blood supply ; pathology ; Cerebral Cortex ; pathology ; Cerebrovascular Disorders ; complications ; pathology ; Cognition ; Corpus Callosum ; pathology ; Diffusion Magnetic Resonance Imaging ; Female ; Hippocampus ; pathology ; Humans ; Male ; Temporal Lobe ; pathology

OBJECTIVETo perform the detection of spermatozoal gene expression in order to accelerate the study of spermatozoal molecular biology.

METHODSTo collect the healthy adults sperm and lymphocytes respectively, and then to extract the total RNAs from them by RNeasy mini kit (QIAGEN) or Trizol reagent. Corresponding cDNAs were produced, digested, ligated, finally labeled with Cy3 (sperm) and CyS (lymphocyte) in the course of RD amplifying reactions. Hybridization with self-made microarrays contained 560 probes was carried out after the labeled cDNAs pured by PCR Product Purification Kit.

RESULTSAmong the 560 probes, 72 genes were up-regulated, 321 genes were down-regulated, the others had no different expression. Furthermore, genes associated with replication, transcription, translation and regulative functions were non-different expression or down-regulated, and those belonged to the spermatogenesis associated, sperm associated antigen were up-regulated, but those involved in the glycolysis were up-regulated, in the oxidative phosphorylation were down-regulated.

CONCLUSIONIt had successfully confirmed that there were a plenty of genes expressed in sperm, furthermore the genes expressed were accorded to spermatozoal functions and characteristics.

Adult ; Down-Regulation ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; Lymphocytes ; metabolism ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; RNA ; isolation & purification ; Spermatozoa ; metabolism ; Up-Regulation

OBJECTIVETo explore the correlation of the mutation of MTCYB and MTATP6 genes in sperm mitochondria with asthenospermia.

METHODSWe extracted mtDNA from 80 semen samples of asthenospermia and 20 of normal sperm motility, amplified the MTCYB and MTATP6 genes by PCR, and analyzed their mutation by sequencing and BLAST matching.

RESULTSThe deletion of both MTCYB and MTATP6 were detected in 20 of the 80 asthenospermia samples, MTCYB deletion in 16 and MTATP6 deletion in 4, accounting for 20% and 5% respectively. Sequencing and BLAST matching revealed G8887A mutation in the MTATP6 gene in the asthenospermia samples, with a mutation rate of 20%, while no regular mutation was noted in MTCYB. Neither significant deletion nor mutation was observed in any of the 20 samples of normal sperm motility.

CONCLUSIONBoth the deletion and mutation of MTCYB and MTATP6 genes in sperm mitochondria might affect sperm motility in adults.

Adult ; Asthenozoospermia ; genetics ; pathology ; Base Sequence ; Cytochromes b ; genetics ; DNA, Mitochondrial ; genetics ; Humans ; Male ; Mitochondrial Proteins ; genetics ; Mitochondrial Proton-Translocating ATPases ; genetics ; Molecular Sequence Data ; Mutation ; Sequence Homology, Nucleic Acid ; Sperm Count ; Spermatozoa ; metabolism ; pathology

Objective To evaluate the MRI findings in patients with general paresis of insane (GPI) to enhance the understanding of MRI diagnosis of this disease. Methods The clinical data and MRI findings of 32 patients with GPI, admitted to our hospital from May 2006 to November 2010, were retrospectively analyzed. Results The MRI findings of GPI were mainly divided into 2 types: cerebral atrophy (n=30) and cerebral swell (n=2). The major MRI findings in the type of cerebral atrophy included white cerebral atrophy in the temporal lobe, the frontal lobe, the hippocampus and the corpus callosum, morphological changes and T2 hyperintensity in the amygdaloid body and the hippocampus, gyral T2 hyperintensity in the cortex and subcortex, and T2 hypointensity in the lenticular nucleus. The MRI findings in the type of cerebral swell manifested as suffused and focal types. Conclusion The MRI findings in GPI are multiple with some characteristic manifestations. Diagnosis must be made through the combination of imaging features with clinical data and related laboratory tests.

The study was purposed to investigate the effect of phosphorylated-chk1 on cell cycle and apoptosis of human erythroleukemic cell line K562 and K562/A02, and to explore the mechanism of chk1 in regulation of drug-resistance of leukemia cells. After treatment with adrimycin for six hours, the cell cycle distribution was detected by flow cytometry; the Chk1mRNA expression was detected by RT-PCR and the Chk1 phosphorylation level was detected by Western blot. Under the condition of down-regulation of Chk1mRNA expression in cells transfected with Chk1 short hairpin RNA, the cell apoptosis rates were detected by flow-cytometry following adrimycin. The results indicated that the proportion of K562/A02 cell line in G2/M phase was (54.12 +/- 0.57)% at 6 hours after drug treatment, significantly higher than that of K562 cell line (36.99 +/- 1.28)%. No evident difference of the Chk1mRNA expression was observed between K562 and K562/A02 cell lines, while elevated Chk1 phosphorylation following DNA damage induced by adriamycin was observed in the K562/A02 cell line (0.79 +/- 0.56), significantly higher than that in K562 cell line (0.27 +/- 1.47). The cell apoptosis rate of the Chk1 shRNA group in K562/A02 cell line was 3.84-fold of blank vector group, but that in K562 cell line was 1.30-fold of blank vector group. It is concluded that the increased chk1 activity that delay the progress of cell cycle are associated with cellular resistance to adrimycin in the K562/A02 cell line.
Apoptosis ; physiology ; Checkpoint Kinase 1 ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; genetics ; G2 Phase ; Humans ; K562 Cells ; Mitosis ; Phosphorylation ; Protein Kinases ; biosynthesis ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; Signal Transduction