Glomerular hypertrophy is the main pathological characteristic in the early stage of diabetic nephropathy (DN), and its regulatory mechanism is closely related to mammalian target of rapamycin (mTOR) signaling pathway activity. mTOR includes mTOR complex 1 (mTORC1) and mTOR complex 2(mTORC2), in which, the upstream pathway of mTORC1 is phosphatidylinositol-3-kinase (PI3K)/serine-threonine kinase(Akt)/adenosine monophosphate activated protein kinase(AMPK), and the representative signaling molecules in the downstream pathway of mTORC1 are 4E-binding proteins(4EBP) and phosphoprotein 70 S6Kinase(p70S6K). Some Chinese herbal extracts could improve cell proliferation via intervening the expressions of the key molecules in the upstream or downstream of PIK/Akt/mTOR signaling pathway in vivo. As for glomerular mesangial cells(MC) and podocyte, mTOR plays an important role in regulating glomerular inherent cells, including adjusting cell cycle, energy metabolism and matrix protein synthesis. Rapamycin, the inhibitor of mTOR, could suppress glomerular inherent cell hypertrophy, cell proliferation, glomerular basement membrane (GBM) thickening and mesangial matrix deposition in model rats with DN. Some Chinese herbal extracts could alleviate glomerular lesions by intervening mTOR signaling pathway activity in renal tissue of DN animal models or in renal inherent cells in vivo and in vitro.
Animals ; Diabetic Nephropathies ; drug therapy ; enzymology ; genetics ; pathology ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hypertrophy ; drug therapy ; enzymology ; genetics ; pathology ; Kidney Glomerulus ; drug effects ; metabolism ; pathology ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; genetics ; metabolism

Objective To evaluate the predictive value of Holter ECG recordings for patients with moderate-severe obstructive sleep apnea and hypopnea syndrome ( OSAHS ) . Methods Holter recordings was performed in 76 patients who were diagnosed OSAHS by polysomnography( PSG) within one month from Jan. 2008 to July 2009 in our hospital. Twenty-eight patients were identified as mild OSAHS (AHI≤20) and forty-eight patients were moderate-to-severe OSAHS ( AHI >20). The indexes of heart rate variability (HRV) , total scores of thirteen sleep apnea risk indexes of Holter recordings and BMI were analyzed by bivariate Logistic regression analysis. Results Clinical features ( eg. Gender, age, complicated with hypertension,coronary heart disease, diabetes mellitus, hyperglycemia, and taken β-blocker) , total scores, the sum of thirteen sleep apnea risk scores collected by Holter recordings (5. 64 ± 2. 33 vs. 6.42 ± 2. 22, respectively,P>0. 05 ) were similar between patients with mild OSAHS and moderate-to-severity OSAHS. VLF/Total Power>70% ,the difference of daytime/nighttime LF Power < -70 and BMI were independent predictors of moderate-to-severe OSAHS with OR 3. 98 (1. 087 - 14. 596), 3. 69 (1. 106 - 12. 285) and 1. 28 (1. 062 - 1. 544), respectively (all P < 0. 05). Conclusions VLF/Total Power and the difference of daytime/nighttime LF Power and BMI could be used as screening parameters to recognize patients with moderate-to-severe OSAHS.

OBJECTIVETo investigate methods of promoting revascularization of tracheal transplantation to increase the length of graft.

METHODSTransfer recombinant plasmid pcDNA3.1/myc-His(-)C-bFGF and pCD(2)-VEGF(121) into rabbit cervical muscle by direct injection of plasmid following electric pulses in vivo. Use histochemistry and immunohistochemistry analysis of muscles injected to show the transferred gene expression and the biological effect. Based on the former experiment, conduct gene therapy to the rabbit tracheal autotransplantation wrapped by cervical combined muscles by injecting plasmid DNA directly, combined with gene sutures following electric pulses. Observe and analyze the effect on trachea viability.

RESULTSThe recombinant plasmid, pcDNA3.1/myc-His(-)C-bFGF and pCD(2)-VEGF(121) was transferred into muscles flap in vivo successfully. The active protein bFGF and VEGF(121) were expressed at high levels. Blood vessels increased significantly in the muscles, and blood circulation was improved by local angiogenesis. Ten rings tracheal autograft wrapped by transgenic muscles integrating with gene structure revascularized completely, and the rabbit survived for a long period of time. There was significant difference between gene therapy group and control group (P < 0.01). There was no significant difference between bFGF gene therapy group and VEGF(121) gene therapy group. Almost rabbits in the control group died of graft necrosis.

CONCLUSIONTracheal grafts revascularization can be established early by the cervical combined muscles flap wrapping associated with single gene therapy. The length of the tracheal can be increased simultaneously.

Animals ; Female ; Fibroblast Growth Factor 2 ; genetics ; therapeutic use ; Genetic Therapy ; Male ; Neck Muscles ; Rabbits ; Surgical Flaps ; Trachea ; blood supply ; transplantation ; Transfection ; Transplantation, Autologous ; Vascular Endothelial Growth Factor A ; genetics ; therapeutic use

Antigens, Viral ; genetics ; metabolism ; Cell Line ; Cell Phone ; Gene Expression Regulation, Viral ; radiation effects ; Herpesvirus 4, Human ; physiology ; radiation effects ; Humans ; Immunohistochemistry ; Viral Proteins ; genetics ; metabolism

Objective The purpose of this study was to approach the relation of SNP43,SNP44 locus, main haplotypes and haplotype combinations with type 2 diabetes mellitus(T2DM).Methods According to the theory and principles of systematic review,data from case-control studies regarding the association between calpain-10(CAPN10) gene and T2DM were derived through electronic search of PubMed and Chinese journals databases.To gain a more precise estimation of the relationship,a stratified Meta-analysis with four subgroups was pertbrmed according to the races.Publication bias Was also assessed.Results The association with T2DM in different races was evaluated.In Mongoloid race,SNP43-G allele,G/G genotype and 111/221 haplotype combination showed notable association with T2DM with Ors (95%CI) as 1.368(1.155-1.620),1.437(1.186-1.741) and 2.762 (1.287-5.927) respectively.In Caucasoid race,SNP44-C allele,111/111 hapotype combination showed strong relationship with T2DM with Ors(95%CI) as 1.144(1.023-1.278),1.291(1.050-1.586) respectively.In Hybrid race,only one positive finding Was obtained which Was SNP44-C allele with OR(95%CI)as 1.653(1.025-2.665).Conclusion SNP43-G allele,G/G genotype,111/221 were risk factors to Mongoloid race.And SNP-C allele,111/111 haplotype combination were risk factors to Caucasoid race,and SNP44-C allele to Hybrid race.

To explore the gene-based principal component logistic regression model and its application in genome-wide association study.Using the simulated genome-wide single nucleotide polymorphism (SNPs) genotypes data,we proposed a practical statistical analysis strategy-'the principal component logistic regression model',based on the gene levels to assess the association between genetic variations and complex diseases.The simulation results showed that the P value of genes in related diseases was the smallest among the results from all the genes.The results of simulation indicated that not only it could reduce the degree of freedom through hypothesis testing but could also better understand the correlations between SNPs.The gene-based principal component logistic regression model seemed to have certain statistical power for testing the association between genetic genes and diseases in the genome-wide association studies.

Objective To compare the incidence of emotional disorder in patients with acute or stable coronary heart disease. Methods A total of 298 patients with suspected coronary heart disease (CHD) were designed into three groups based on of coronary angiography results: acute coronary syndrome (ACS, n=128), stable angina pectoris (SAP, n=108) and non-CHD (n=62). All patients were evaluated by Zung Self-rating Depression Scale (SDS), Zung Self-rating Anxiety Scale(SAS) and Hamilton Depression Rating Scale (HRSD) for depression and anxiety before coronary angiography (CAG), 3 days after CAG, and 1 day before discharge. Results Incidences of depression and anxiety were significantly higher the ACS group (65.6% and 78.9% before CAG; 60. 9% and 70. 3% 3 days post CAG; 45.3%and 64. 8% before discharge) compared patients with SAP (18.5% and 26.9% before CAG; 17.6% and 28.7% 3 days post CAG; 15.7% and 26.9% before discharge, all P <0.05 vs. ACS) and non-CHD patients (32.3% and 25.8% before CAG; 27.4% and 24.2% 3 days post CAG; 29.0% and 30.6% before discharge, all P<0.05 vs. ACS) while the depression and anxiety incidences were similar between patients with SAP and non-CHD in this cohort (P>0.05). Conclusion Emotional disorder is common in patients with suspected heart diseases, especially in patients with ACS. Psychological distress of patients with suspected heart disease should be evaluated and treated.

This study was aimed to investigate the effects of xenogeneic antigen neu-Fc in combination with the recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF) and Bacillus Calmette-Guerin (BCG) on the regulation of Th1 and Th2 immune response in vitro. The rat neu L2-S2 domain was engineered as a chimeric protein with human IgG Fc. The eukaryotic expression vector was constructed. The recombinant protein was stably expressed in CHO cells and purified by rProtein A Sepharose Fast Flow column. The recombinant protein was identified by SDS-PAGE and Western blot. Peripheral blood mononuclear cells (PBMNCs) were obtained by means of standard Ficoll separation from the blood of healthy donors. Neu-Fc-induced PBMNC proliferation was tested by MTT. The production of IL-12 and IL-10 was measured by ELISA. The results showed that the level of IL-12 decreased and IL-10 increased after PBMNCs were incubated with MCF-7 cultural supernatant. 10 nmol/L neu-Fc strongly induced the cell proliferation. Compared with neu-Fc or GM-CSF or BCG treatment alone, neu-Fc in combination with GM-CSF and BCG significantly stimulated IL-12 production and inhibited IL-10 production (p < 0.01). It is concluded that the neu-Fc can stimulate the proliferation activity of PBMNCs. neu-Fc, GM-CSF and BCG costimulation efficiently induces Th1 immune response.
Animals ; BCG Vaccine ; immunology ; CHO Cells ; Cricetinae ; Cricetulus ; Granulocyte-Macrophage Colony-Stimulating Factor ; immunology ; Humans ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism ; Rats ; Recombinant Proteins ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology

Interferon gamma-inducible protein 10, a member of the family of CXC chemokines, is secreted by interferon gamma-stimulated, monocytes, endothelial cells and keratinocytes. Interferon gamma-inducible protein 10 plays an important role in recruiting activated T cells into sites of tissue inflammation. In this experiment, PCR products of Interferon gamma-inducible protein 10 were cloned into prokaryote expression vector pET 32(a) to generate recombinant pET-IP10 with S-Tag at the N-terminus, and expressed successfully in E. coli BL21 (DE3). The total expressed products amounted to 25.3% in all bacterion proteins. pET-IP10 mainly formed inclusion body in E. coli. Soluble recombinant protein accounted for 20% among IP-10 fusion protein. The soluble recombinant proteins were purified by using S-Tag affinity chromatography effectively with purity of over 90%. The chemotaxis biological activity of purified Interferon gamma-inducible protein 10 could specifically exhibit the directional migration of stimulated T cells at concentration of 100 ng/ml. The results indicated that the strategy we used in this experiment was effective for recombinant Interferon gamma-inducible protein 10 production with biological activity.
Chemokine CXCL10 ; biosynthesis ; Escherichia coli ; metabolism ; Genetic Vectors ; Humans ; Recombinant Proteins ; biosynthesis ; T-Lymphocytes ; cytology ; Thioredoxins ; biosynthesis

OBJECTIVEA prospective study was conducted to probe into the relationship between arterial oxygen partial pressure (PaO2) and brain injury during cardiopulmonary bypass (CPB) in infants with cyanotic congenital heart disease (CHD).

METHODEnrolled in the study were 45 cyanotic infants, who were less than three years old and underwent corrective cardiac surgery from August 1(st), 2010 to January 31(st), 2011 at Guangdong General Hospital. All the infants had a pulse oxygen saturation (SpO2) lower than 85% and were randomly allocated into three groups by a specific computer program. In controlled group 1 (G1 group), PaO2 levels were controlled at 80 - 120 mm Hg (1 mm Hg = 0.133 kPa) during CPB; in controlled group 2 (G2 group), PaO2 levels at 120 - 200 mm Hg during CPB; while in uncontrolled group (G3 group), PaO2 levels were at 200 - 400 mm Hg during CPB. Blood samples were collected just before starting CPB, at the end of CPB, and at 3 h, 5 h, and 24 h after CPB (T1, T2, T3, T4, T5) for the determination of serum concentrations of protein S100β, neuron specific enolase (NSE), and adrenomedullin (ADM) by ELISA.

RESULTProtein S100β rose significantly after starting CPB. In group G3, it reached a peak of (699 ± 139) ng/L by the end of CPB, significantly higher than those in groups G1 and G2 [(528 ± 163) ng/L and (585 ± 155) ng/L], and was positively correlated with PaO2 levels (r = 0.526, P < 0.01). NSE levels of group G1 were continuously rising after starting CPB and reached significantly high levels at 3 h or 5 h after CPB [(12.2 ± 3.4) µg/L and (12.3 ± 3.7) µg/L], while those of group G2 rose significantly during CPB [(10.9 ± 4.8) µg/L] and even higher at 3 h or 5 h after CPB [(12.6 ± 5.1) µg/L and (13.2 ± 5.4) µg/L]. NSE levels of group G3 rose significantly during CPB and maintained at a high level [(12.2 ± 5.7) µg/L] afterwards. There was no significant difference in serum ADM concentrations among different time points in each group and among these three groups. All the infants were discharged from the hospital without any obvious nervous symptom and sign.

CONCLUSIONHigh PaO2 during CPB in infants with CHD might cause an increase of serum protein S100β and NSE, indicating that brain injury might become worse with a higher PaO2 and might be positively correlated with PaO2 during CPB.

Cardiopulmonary Bypass ; Child, Preschool ; Cyanosis ; Female ; Heart Defects, Congenital ; blood ; physiopathology ; surgery ; Humans ; Infant ; Male ; Nerve Growth Factors ; blood ; Oximetry ; Oxygen ; blood ; Partial Pressure ; Phosphopyruvate Hydratase ; blood ; Prospective Studies ; S100 Calcium Binding Protein beta Subunit ; S100 Proteins ; blood ; Serum