Establishment of immune liver fibrosis models in cynomolgus monkeys / 中国免疫学杂志

Objective:To establish liver fibrosis models in cynomolgus monkey using calf serum.Methods:Healthy cynomolgus monkeys were randomly divided into control group and model group,and there were 6 animals in each group.Model group were treated with calf serum subcutaneously at the dose of 0.4 ml/kg,and control group were received the same dose of normal saline.Calf serum will be administrated twice a week.Results:After 8 weeks of administration,the serum HA,PC Ⅲ,Ⅳ C,TGF-β1 and α-SMA in model group were increased differently when compared with modeling before(P＜0.05,P＜0.01),and the serum HA and PC Ⅲ in model group were also increased significantly when compared with control group.After 12 weeks of administration,the serum Ⅳ C,TGF-β1 and α-SMA in model group were increased significantly compared with control group(P＜0.05),and the serum LN and TCH in model group were increased obviously when compared with modeling before(P＜0.05),the levels of serum AST and ALT in model group were also increased distinctly as compared with control group and modeling before(P＜0.05),while the serum ALB in model group was decreased significantly when compared with modeling before(P＜0.05).At week 16,the serum LN,TCH and TG in model group were increased significantly when compared with control group(P＜0.05);while the serum ALB in model group was decreased distinctly when compared with control group(P＜0.05);the strong echo were showed in liver of model group by using ultrasonography;pathological examination showed that there were necrosis and monocyte infiltrating in liver of model group,and collagen fiber proliferation were observed in portal area of model group.Conclusion:Liver fibrosis models can be established successfully in cynomolgus monkeys by subcutaneously injecting calf serum for 16 weeks.

Reduced estimated glomerular filtration rate and proteinuria are associated with increased cardiovascular events rate in octogenarian population / 中华心血管病杂志

<p><b>OBJECTIVE</b>To determine the relationship between estimated glomerular filtration rate (eGFR) and proteinuria with cardiovascular events in subjects aged 80 years or older.</p><p><b>METHODS</b>Data for this retrospective prognostic study were drawn from the patient database for routine checkup in Beijing hospital between January 2001 to December 2001. Baseline eGFR and proteinuria were evaluated in 340 subjects [mean age: (85.6 ± 4.0) years]. eGFR was calculated using the modified abbreviated MDRD equations based on the Chinese chronic kidney disease patients. The subjects were divided into normal renal function group and reduced renal function group (eGFR <60 ml·min(-1)·1.73 m(-2)). The subjects were divided into subjects without proteinuria and subjects with proteinuria group. Cardiovascular events included cardiovascular death, nonfatal myocardial infarction, nonfatal stroke.</p><p><b>RESULTS</b>The proportion of reduced renal function was 36.8% (125/340). The proportion of proteinuria was 10.3% (35/340). The proportion of reduced renal function or proteinuria was 41.8% (142/340). Follow-up time was 79 months (40-114 months). Cardiovascular events rate was significantly higher in reduced renal function group than in normal renal function group [37.6% (47/125) vs. 26.2% (55/210), P < 0.05 ] and in proteinuria group than in without proteinuria group [50.0% (17/34) vs. 28.2% (85/301), P < 0.01 ]. Cox multivariate analysis revealed that both eGFR (HR = 0.978, 95%CI:0.961-0.994, P < 0.05 ) and proteinuria (HR = 2.049, 95%CI:1.132-3.709, P < 0.05) were independent risk factors for cardiovascular events after adjusting for age, gender, triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, uric acid, hypertension, coronary heart disease, diabetes mellitus.</p><p><b>CONCLUSIONS</b>Reduced eGFR and presence of proteinuria are independent risk factors for cardiovascular event in subjects aged 80 years or older. eGFR and proteinuria can thus be used for cardiovascular event risk stratification in subjects aged 80 years or older.</p>

The inhibitory effect of capsaicin on streptozocin-induced apoptosis of rat retinal cells / 中华实验眼科杂志

Background Diabetes mellitus (DM) can provoke the apoptosis of retinal cells and downregulate the expression of calcitonin gene related peptide (CGRP) in the retina.Capsaicin promotes the release of CGRP and elicits protective effects on human organs.However,whether CGRP protects retinal cells in diabetic retinopathy (DR) is still unclear.Objective The study was designed to examine the effect of capsaicin on the apoptosis of retinal cells in diabetic rats and its relationship with CGRP.Methods Forty clean healthy adult male Sprague-Dawey rats were randomly divided into the diabetes group,capsaicin pretreated group,streptozocin (STZ)control group,capsaicin control group and plain control group,with 8 rats per group.The diabetic model was established by the intraperitoneal injection of 60 mg/kg in all rats except those of the plain control group.0.4 mL of a 1％ capsaicin injected at 20 mg/kg was subcutaneously injected for 3 consecutive days prior to model establishment in the capsaicin pretreated group,after which 1.2 mL of STZ was intraperitoneally injected on the fourth day.Rats from the STZ control group were administered intraperitoneally 1.2 mL of 0.1 mol/L,pH 4.5,citrate buffer.The capsaicin control group received subcutaneous injections of 0.4 mL of 1％ capsaicin at 20 mg/kg for 3 consecutive days,after which 1.2 mL of 0.1 mol/L,pH 4.5,citrate buffer was administered intraperitoneally.The rats were sacrificed at the tenth week after model establishment and retinal specimens were prepared for the apoptosis assay by TUNEL staining and the quantitative analysis of caspase-3 activity.Expression of CGRP in the retina and serum was detected using ELISA.The use of experimental animals followed the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results Retinal cell apoptosis was mainly localized to the retinal ganglion cell (RGC) layer.The apoptosis rate of RGCs was (43.4±5.0)％ in the DR model group and (30.0±5.1)％ in the capsaicin pretreated group,showing a significant difference (t =5.930,P＜0.01).Compared with the DR model group and capsaicin pretreated group,the apoptosis rates of the DR control group (12.4±9.9) ％ and the capsaicin control group (17.6-±6.1) ％ were significantly lower (t =8.800,t =4.925,P＜0.01).The apoptosis rate of the plain control group was (16.2±6.9)％,exhibiting significant differences in comparison with the DR control group and capsaicin control group (t =-0.989,t =0.951,P＞0.05).The specific activity of caspase-3 was (2.19±0.86) in the DR model group and (1.96±0.56) in the capsaicin pretreated group,presenting a significant difference (t =-0.515,P＜0.05).Those of the DR control group and capsaicin control group were (1.47±0.14) and (0.74±0.27),respectively,with considerable decline in comparison with the DR model group and capsaicin pretreated group (t=2.142,t=2.797,P＜0.05).The retinal and serum CGRP levels were (424.4±44.2)and (148.8±39.1) ng/L,respectively,displaying significantly lower levels than (543.2±74.4) and (237.5±78.7) ng/L (t =3.070,2.359,P＜0.05) from the capsaicin pretreated group.Conclusions Apoptosis of retinal ganglion cells occurs in the STZ-induced diabetic rats.Pretreatment of capsaicin reduces retinal cell apoptosis,which may be associated with an increase of CGRP in the retina.

Role of endogenous calcitonin gene related peptide in stress-induced retinal cell apoptosis / 中华实验眼科杂志

Background Acute stress can provoke the apoptosis of retina cells and induce increasing expression of calcitonin gene related peptide(CGRP)in retina.However,the role of CGRP in pathology of the stressinduced apoptosis of the retina ceils is still elusive.Objective The aim of this study was to investigate the effects of endogenous CGRP on retinal cell apoptosis induced by stress of acute myocardial ischemia after coronary artery occlusion in rats. Methods The acute myocardial ischemia model was established by ligating the left anterior descending branch of coronary artery in 12 male Sprague-Dawley rats.The rats were randomized into CGRP8-37 injection group and normal saline injection group,6 rats 12 eyes for every group.CGRP8-37(10-7 mol/L),a specific antagonist of CGRP receptor,was intravenously injected in CGRP8-37 group by caudal vine at 15 minutes prior to the coronary artery occlusion,and the equivalent amount of normal saline was used at the same fashion in normal saline group.The retinal samples of the rats were collected at 3 hours after coronary artery occlusion for TUNEL staining and caspase-3 activity detection respectively. Results The cellular displacement was observed in inner and outer nuclear layer,and vacuolar degeneration of retinal ganglion cells was found in the coronary artery occlusion animals.The total apoptosis index of retinal cells in CGRP8-37 group was significantly higher than that in normal saline group (42.8％±2.8％ vs 37.5％±2.9％,t=-3.244,P<0.01).The retinal capase-3 activity was significantly enhanced in the CGRP8-37 group compared with saline group(11.3±3.1 fold vs 4.9±1.2 fold,t=-4.603,P<0.01)at 3 hours of coronary artery occlusion.Conclusion The results suggest that the endogenous CGRP may play an anti-apoptotic role in the stress.induced retinal cell injury.

Regulation of NF-κB signal transduction pathway on cytokines in cultured nasal epithelium / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the activation and regulation of nuclear factor-kappa B (NF-κB) on transcription of cytokines in cultured lipopolysaccharide (LPS)-induced nasal epithelial cells.</p><p><b>METHODS</b>Normal sphenoid mucosa epithelium from 11 patients who accepted pituitary tumor surgery via trans-sphenoid approach was separated and cultured without serum. The epithelium of the third or the forth passage was induced with LPS. Wedelolactone, a blocking agent of NF-κB was used at the same time. An electrophoretic mobility shift assay was used to detect DNA-binding activity of NF-κB. The reverse transcription-polymerase chain reaction was used to evaluate mRNA of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), IL-5, IL-6, IL-8, granulocyte-macrophage colony stimulating factor (GM-CSF), regulated on activation, normal T expressed and secreted (RANTES), eotaxin, eotaxin-2, vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Statistical analysis was performed using SPSS17.0 software.</p><p><b>RESULTS</b>DNA binding activity of NF-κB and mRNA of IL-1β, IL-8 and COX-2 increased in cultured LPS-induced nasal epithelial cells (relative values were 1.013 ± 0.144, 0, 0, 0 respectively in control group and relative ones of LPS-induced were 2.050 ± 0.305, 1.057 ± 0.041, 0.950 ± 0.042, 0.117 ± 0.012 respectively). There was significant difference between the control group and LPS-induced nasal epithermal cells group (P values were 0.004, 0.000, 0.000, 0.000 respectively). Corresponding expression of NF-κB, IL-1β, IL-8 and COX-2 decreased after the addition of Wedelolactone (relative values were 0.917 ± 0.188, 0.180 ± 0.008, 0, 0 respectively). There was significant difference between the LPS-induced nasal epithelial cells group and the Wedelolactone-addition group (P values were 0.002, 0.000, 0.000, 0.000 respectively). But the expression of mRNA of other factors were 0 in all groups.</p><p><b>CONCLUSIONS</b>The NF-κB signal transduction pathway was involved in the transcriptional regulation of IL-1β, IL-8 and COX-2 in cultured LPS-induced nasal epithelial cells.</p>

Magnetic resonance imaging of the upper airway structure of children with sleep disordered breathing / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the upper airway structure of sleep-disordered breathing children.</p><p><b>METHODS</b>Seventy three children with obstructive sleep apnea hypopnea syndrome (OSAHS), 53 children with primary snoring (PS) and 40 control subjects underwent pharyngeal magnetic resonance imaging (MRI). Upper airway structure images were analyzed and measured.</p><p><b>RESULTS</b>The cross-section area of the nasopharyngeal and palatopharyngeal airway in subjects with OSAHS and PS are smaller (P < 0.01) than that of the control group. The cross section area of OSAHS patients are smaller than that of PS subjects (P < 0.01). The above parameter of oropharyngeal airway in OSAHS patients is smaller than that of control group (P < 0.01), but no statistic difference compared with that of PS subjects. The cross-section area and length of the adenoid in OSAHS group are bigger and longer than that of PS group (P < 0.01) and bilateral tonsils are larger (P < 0.01); in OSAHS patients the cross-section area of the soft palate is larger and the length of the soft palate is longer (P < 0.01) than that of PS group, while this parameter of PS group is similar to that of the control group. And the maximum width of the soft palate, the cross-section area of bilateral fat pad, bilateral pterygoid and tongue are similar among OSAHS, PS and the control group. The skeletal measurement: the length of H-C2C3 in subjects with OSAHS is longer (P < 0.01); The angle(alpha) in OSAHS patients is smaller (P < 0.01) than that of other 2 groups. The angle (beta), the cross-section area of the mandible, the spine-clivus oblique, the length of the hard palate and the distance of the mandible are similar among the three groups.</p><p><b>CONCLUSIONS</b>In children with OSAHS or PS, the upper airway is restricted by both the adenoid and tonsils; however, the soft palate is also larger in OSAHS, adding further restriction. Otherwise, downward movement of the hyoid bone and decreasing of the angle (alpha) in OSAHS influence laryngopharynx airway. MRI is of clinical significance for evaluating OSAHS children's upper airway.</p>

Effect of BMP-7 on the transdifferentiation of cultured human tubular epithelial cell induced by TGF-beta1 / 中国医学科学院学报

<p><b>OBJECTIVE</b>To observe the effect of bone morphogenetic protein-7 (BMP-7) on the transdifferentiation of cultured human tubular epithelial cell (HKC) induced by TGF-beta1 and to elucidate its possible mechanism.</p><p><b>METHODS</b>The cultured HKC cells were divided into 5 groups: serum-free group (negative control); single TGF-beta1 treated group (positive control); single BMP-7 treated group; combined TGF-beta1 and BMP-7 treated group; and BMP-7 pre-treated group. Expression of keratin of HKC cells was assessed by indirect enzyme immunohistochemistry (IEI), expression of alpha-smooth muscle actin (alpha-SMA) and E-cadherin by immunohistological method, percentage of alpha-SMA positive HKC cells by flow cytometry, and mRNA expression of alpha-SMA, TGF-beta1, and TGF-beta type II receptor by reverse transcription PCR.</p><p><b>RESULTS</b>The expression of alpha-SMA and the percentage of alpha-SMA positive HKC cells markedly increased after having been treated by TGF-beta1 while the expression of E-cadherin and keratin decreased. In the group pre-treated with BMP-7 (50 ng/ml) and then added with TGF-beta1 (8 ng/ml), expression of alpha-SMA was significantly lower than in the positive control group, while expression of E-cadherin and keratin significantly higher than in the positive control group. Measurement of the percentage of alpha-SMA positive HKC found significant deference between the combined TGF-beta1 and BMP-7 treated group and the positive control group (9.7% vs 19.8%; 5.8% vs 19.8%; P < 0.05). Significant difference existed between the BMP-7 (50 ng/ml) pre-treated group and the positive control group (8.7% vs 19.8%, P < 0.05). mRNA expression of alpha-SMA was measured by RT-PCR and the results showed that it significantly decreased in the group treated or pre-treated with BMP-7 (50 ng/ml) (15% and 12% of the results in the positive control group, respectively). The mRNA expression levels of both TGF-beta1 and its type II receptor significantly decreased (28% and 19%; 47% and 36%, compared with the positive control group, respectively).</p><p><b>CONCLUSION</b>Transdifferentiation of cultured renal epithelial cell induced by TGF-beta1 can be inhibittd by certain levels of BMP-7, cultured together with TGF-beta1 or pretreated. BMP-7 can prevent and inhibit the mRNA expression of TGF-beta1 and its type II receptor, which may be an important mechanism by which BMP-7 inhibit the transdifferentiation of renal tubular epithelial cell.</p>

Evaluation and comparison of various renal function tests in elderly patients with chronic kidney disease / 中华老年医学杂志

Objective To find out whether the levels of serum creatinine and urea,creatinine clearance rate and GFR estimated by using different equations correlate with the exact measurement of GFR with ~(99m)Tc-DTPA in elderly patients with chronic kidney disease.Methods Renal function was determined by using ~(99m)Tc-DTPA technique in 83 elderly patients aged 65—96 years.Blood and urine tests were done at the same time.Estimated clearance was determined by using the equations of Cockcroft-Gauh,MDRD(modification of diet in renal disease)predigestion and MDRD seventh,and the correlation coefficients between the above parameters and ~(99m)Tc-GFR were determined.Veracity for the diagnosis of renal dysfunction was calculated by using receiver operating characteristic(ROC) curves.Results Cockcroft-Gault formula(r=0.833),MDRD predigestion formula(r=0.811), MDRD seventh formula(r=0.803),creatinine(r=-0.672),Ccr(r=0.632),urea(r=-0.612), and albumin(r=0.444)were correlated significantly with ~(99m)Tc-GFR in this sequence.If GFR 30, 40,50,60ml/min were defined respectively as renal impairment thresholds and areas under the curve (AUC)were compared,the best parameter for estimation of GFR was the Cockcroft-Gault formula. Then MDRD predigestion formula and MDRD seventh formula,creatinine and Ccr were less accurate in all different renal dysfunction thresholds.Conclusions The Cockcroft-Gault equation seems to be the best formula available for GFR estimation,the two equations of MDRD are the second,creatinine is the third,Ccr and urea are the last.

Changes of tumor necrosis factor-? levels in serum and cerebrospinal fluid of childhood acute leukemia before and after treatment / 实用儿科临床杂志

Objective To explore the changes of tumor necrosis factor-?(TNF-?) in serum and cerebrospinal fluid(CSF) of children with acute lymphoblastic leukemia (ALL) and acute myelogenous leukemia(AML) and its clinical significance.Methods TNF-? in serum and CSF were measured by radioimmunoassay and CSF samples were obtained from 31 cases of childhood acute leukemia before treatment, on complete remission(CR), and continuous CR.Results Serum TNF-? was in ALL and AML before treatment [(24.35?4.84) pmol/L and(28.65?5.12) pmol/ L],which were significantly higher than those of healthy controls[(11.2 8? 1.69) pmol/L, P