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This study was aimed to investigate the effect of Yi-Qi Wen-Yang (YQWY) and Huo-Xue Li-Shui (HXLS) decoction on changes of insulin resistance (IR) model and the myocardial tissues of rats with congestive heart failure (CHF) pathology. CHF rat models were established on Wistar male rats through injection of doxorubicin hydrochlo-ride into rat tail vein. Wistar male rats models were randomly divided into the model group, western medicine group, low dose decoction group, middle dose decoction group, and high dose decoction group. After 4-week gavage, 3 mL vein blood was taken from the angular vein sinus for the determination of blood glucose and serum insulin, and the calculation of IR. Finally, the rats were sacrificed. And then, the heart was removed to make HE slice and observe the pathological change of myocardium. The results showed that compared with the model group, YQWY and HXLS decoction can improve the IR level among CHF rats (P<0.05). Among them, the effects of the high dose and middle dose group were obvious. At the same time, this decoction can improve the myocardial cells in CHF rats in myocar-dial cells of the high dose group. And its morphology change was close to the digoxin group. It was concluded that YQWY and HXLS decoction can reverse IR and improve ventricular remodeling among CHF rats to a certain extent.
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This study was aimed to observe the protective effect of astragalus on bones of vitamin D deficiency rat model. A total of 30 male SD rats were randomly divided into 3 groups, which were the control group, model group, and the astragalus group. The experimental period was 8 weeks. After the experiment, enzyme immunoassay was used for the detection of serum 25 hydroxy vitamin D3 [25 (OH)VD3], fibroblast growth factor-23 [FGF-23], Klotho, and HE staining of femur. The results showed that compared with the normal control group, the vitamin D deficiency rat model group had a decrease in both serum 25 (OH)VD3 and Klotho, and a increase in FGF-23, which meant the ex-istence of osteoporosis. Compared with the model group, the astragalus group had a decrease in both the serum 25 (OH)VD3 and FGF-23, and a increase in Klotho, which meant the osteoporosis of the astragalus group had been im-proved significantly. It was concluded that astragalus can regulate FGF-23 and Klotho in vitamin D deficiency rats in order to have a protective effect for bones.
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[Objective] To study the methylation status of CD40L gene regulatory regions in peripheral blood CD4+ T cells from patients with systemic sclerosis (SSc).[Methods] Peripheral blood mononuclear cells were isolated from the venous blood of 21 SSc patients and 20 healthy controls by density gradient centrifugation.CD4+ T cells were separated by using magnetic beads.Genomic DNA was isolated from the CD4+ T cells and treated with sodium bisulfite.Nested PCR was perfonned to amplify the desired regulatory sequences (including the promotor and enhancer) of CD40L,and the amplicons were transformed into the Escberichia coli DH5α.Subsequently,8 independent clones were selected and sequenced for each of the amplified fragments.[Results] In healthy female controls,half of the cloned fragments of CD40L regulatory sequences were unmethylated,and the other half were methylated.The mean methylation levels of CD40L promoter and enhancer from female SSc patients were significantly lower than those from healthy female controls (both P ＜ 0.01 ).Almost all of the cloned fragments of CD40L promoter and enhancer were unmethylated in healthy male controls and male SSc patients,with no significant difference in the methylation level between male SSc patients and healthy controls (both P ＞ 0.05,respectively).[Conclusion]s There is a low methylation level of CD40L regulatory elements on the inactive X chromosome in female SSc patients,which may contribute to the CD40L overexpression in CD4+ T cells.
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Objective To investigate the expression levels of CD40L in CD4 + T cells from systemic sclerosis (SSc) patients.Methods PBMC ( peripheral blood mononuclear cells) cells were isolated from the peripheral venous blood of SSc patients (16females,10males) and healthy donors (15females,10males) by density gradient centrifugation.CD4 + T cells were isolated using magnetic beads.mRNA levels of CD40L in CD4 + T cells were measured by real-time quantitative polymerase chain reaction (RTPCR).Flow cytometric analysis was used to detect the CD40L protein on the surface of CD4 + T cells.Results Both CD40L mRNA and protein expression in CD4 +T cells was significantly elevated in female SSc patients compared with female healthy controls [5.61 ± 1.86 vs 2.80 ±0.94,P ＜0.01 ; (6.70 ±3.55)％vs (2.37 ± 1.39)％,P ＜ 0.05,respectively].No significant increase in CD40L mRNA and protein expression was observed in male SSc patients compared with male controls [2.59 ± 0.89 vs 1.92 ± 0.56,P ＞0.05; (2.06±1.09)％ vs (2.13±0.87)％,P ＞0.05,respectively].Conclusions CD40L was overexpressed in CD4 + T cell of female patients but not males,which maybe one of the important reasons for female susceptibility to SSc.
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Objective To explore the changes and clinical significance of serum cystatin-C, β2- mieroglobulin (β2-MG) and renal hemodynamic in children with Henoch-Schoenlein purpura(HSP). Method Forty-six patients with HSP are in HSP group, 40 healthy children are in control group. Serum cystatin-C was determined by enzyme-linked immunosorbent assay, β2-MG was detected by radioimmunoassay, renal hemodynamie was detected by colour Doppler ultrasound. Results Serum cystafin-C and β2-MG in HSP group [(3.96±1.52 ), (2.74±0.82)mg/L] were higher than those in control group [(1.67±0.61), (1.89±0.47)mg/L] (P<0.01). Frequenee spectra showed high velocity and resistance, and maximum crest flow rate[(1.068±0.348) m/s] and resistance index (0.894±0.125) in systolic phase of main renal arteries were obviously higher in HSP group than those in control group [(0.859±0.357) m/s and 0.726±0.078] (P<0.05). Conclusions The level of serum cystatin-C and the change of renal hemedynamie can act as the significant indicators of early diagnosis of HSP nephritis.
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Objective To detect the alteration of circulating CD34 + cell level in patients with the In'st-ever nonlacunar stroke and to investigate the prognostic value of CD34 +cell level. Methods 1he level of the circulating CD34+ cells were examined using flow eytometry in 119 patients with nonlacunar infarction. 1he patients were divided into group A (higher than average) and group B (lower than average) according to the detected values, The National Institutes of Health Stroke Scale (NIHSS) scores on admission and the modified Rankin Scale (mRS) scores at 3 months after the onset in the two groups wre compared. Results The level of CD34 + cells in group A was significantly higher than that in group B (0. 048 ± 0. 001 versus 0. 032 ± 0. 002, P < 0. 05 ), however, there was no significantly difference in NIHSS sores between the two groups. One week after admission, the increased level of CD34 + cells in group A was significantly higher than that in group B (0. 001 ±0. 003 versus - 0. 005 ± 0. 000, P < 0. 05 ). Three months after the onset of symptoms, the average mRS score in group A was significantly superior to that in group B (2. 98 ± 1.14 versus 3.25 ± 1.39, P <0. 05). 1he level of circulating CD34 + cells at the initial onset of symptom was negatively correlated with the mRS score at 3 months after stroke (r = -0. 48, P <0. 05). Conclusions The higher level of circulating CD34+ cells had better prognosis in patients with cerebral infarction. The level of circulating CD34+ ceils may be used as a prognostic indicator in patients with cerebral infarction.
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BACKGROUND: Lung fibroblasts are believed to play an important role in lung tissue repair and regenerative process. The differentiation potential of lung fibroblasts is not known very well.OBJECTIVE: To investigate the multi-differentiation capacity of human lung fibroblasts.DESIGN: An observational comparative experiment.SETTING: Department of Respiratory Medicine, Beijing Shijitan Hospital and Central Laboratory of the First Hospital of Tsinghua University.MATERIALS: This study was performed at the Central Laboratory of the First Hospital of Tsinghua University from March 2006 to October 2006. Human adult lung fibroblasts were isolated as primary cultures from resected lung of patients with lung cancer.The study was approved by hospital's Medical Ethics Committee, and informed consent was signed. Dulbecco's modified Eagle's medium (DMEM), fetal calf serum (FCS), trypsin- ethylenediamine tetraacetic acid (EDTA), naphthol AS-MX phosphate and Fast Red TR were purchased from Sigma, USA. Mouse anti-human osteopondn antibody was from R&D Systems China Co., Ltd,Polyvinyl difluoride (PVDF) membranes were from Bio-Rad Laboratories Inc, Hercules, CA.METHODS: Human adult lung fibroblasts were isolated as primary cultures. Human lung fibroblasts were cultured in an osteogenic medium (containing 109-10-5 mol/L dexamethasone, 50 mg/L vitamin C and 10 mmol/L β-glycerophospate) and adipogenic medium (containing 15% horseurm, 10-8 mol/L dexamethasone and 10 mg/L insulin), or control medium (10% FCS).Ostcoblasts were detected by alkaline phosphatese (ALP) and calcium salt staining. The expression of osteopontin was measured by Western blotting. Oil Red-O staining was used for identification of mature adipocytes.MAIN OUTCOME MEASURES: The differentiation of lung fibroblasts induced by osteogenie medium; The differentiation of lung fibroblasts induced by adipogenic medium.RESULTS: With the induction of ostcogenic medium for 2 weeks, the differentiation of lung fibroblasts was induced by osteogenic and adipogenic medium, respectively. The expression of ALP and osteopontin was increased and the deposition of calcium salt was detected. After lung fibroblasts were cultured in adipogenic medium for 14 days, part of the cells gradually experienced morphological change from original spindle into oval shape. Oil Red -O staining indicated lipid drops accumulation within cytoplasm.CONCLUSION: Under certain condition, human long fibroblasts could differentiate into osteoblasts or adipocytes that were characterized by bone marrow mesenchymal stem cells.
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Objective To investigate the differentiation potential of human lung fibroblasts.Methods The study was performed in First Hospital of Tsinghua University from March 2006 to October 2006.Human lung fibroblasts were cultured for 2 weeks in an osteogenic medium[containing 10% fetal calf serum (FCS)and 50 g/mL Vitamin C and 10 mmol/L ?-glycerophospate],adipogenic medium(containing 15% horse serum,10-8 mol/L dexmethasone and 10 mg/L insulin),or control medium(10% FCS).Osteoblasts were detected by alkaline phosphatese and calcium salt staining.The expression of osteopontin was measured by Western blotting,Oil Red -O staining for identification of mature adipocytes.Results With the induction of osteogenic medium for 2 weeks,the expression of alkaline phosphatese and osteopontin was increased and the deposition of calcium salt was detected.Mature adipocytes formed after culture with adipogenic medium for 2 weeks.Conclusion Under certain condition,human lung fibroblasts can differentiate into osteoblasts or adipocytes,which were characterized by bone marrow mesenchymal stem cells.