ObjectiveTo determine whether there is correlation between the pathological classification and urinary level of transforming growth factor(TGF)-β1,macrophage inflammatory protein (MIP)-1α,and Neutrophil gelatinase-associated lipocalin (NGAL) of lupus nephritis.MethodsELISA was used to test the levels of urinary level of TGF-β1,MIP-lα and NGAL.The correlation between these three urinary markers and the pathological classifications,Austin score,histological semi-quantitative score and clinical data were analyzed.Comparisons between groups were performed with t-test,ANOVA and X2 test.Correlation analysis was performed with Pearson analysis.Results①There was significant difference in urinary TGF- β1,MIP1α,and NGAL levels when compared the lupus nephritis group,SLE patients without renal damage group and the normal control group [TGF-β1(351±219),(92±60),(74±29) pg/ml],[MIP-1α(18.0±15.5),(8.5±2.3),(7.1±1.9) pg/ml],[NGAL (1.104±0.519),(0.181±0.030),(0.146±0.024) ng/ml],while there was no significant difference between lupus nephritis (LN) group and patients with other glomerulonephritis.② There was significant difference in the three urinary markers when stratified the LN patients based on the pathological classifications,however,there was no significant difference between the LN group and other glomerulonephritis group.③ Analysis of the correlation between the three urinary markers and semiquantitative histopathological score of the LN patients showed that the urinary level of TGF-β1 and MsMI,GCI,TCI,VCI was closely related to each other,the urinary MIP-1α was related to and endol,and the dGAI and VCI was closely related, while the urinary NGAL was closely related with endol,dGAI and MsHI.The correlation analysis between the three urinary markers and acute and chronic pathological index score of the LN patients showed that TGF- β1 was correlated with the chronic index(r=0.89,P＜0.01 ),the MIP-lα and NGAL were significantly correlated with the activity index(r=0.71,P＜0.01 ; r=0.60,P＜0.01 ).Conclusion There is a positive correlation between the urine TGF-β1 level and chronic kidney disease.The urinary MIP-lαand NGAL are associated with active renal damage.

Objective To investigate the effects of angelica solution on chronic hypoxic pulmonary hypertension in rats.Methods Thirty SD rats were randomized into normoxic group,hypoxic group and angelica solution-protected group.The model of rat chronic hypoxic pulmonary hypertension was made by method of isobaric hypoxia.Angelica solution were injected before hypoxia,while the other two groups were injected normal saline.After 28d of hypoxia,pulmonary artery pressure were measured.Expressions of proliferating cell nuclear antigen (PCNA) and inducible nitric oxide synthase (iNOS) in pulmonary artery were detected by immunohistochemical staining.The index of wall thickness of rat pulmonary arteriole-percentage of the wall area in the total vascular area(wA%) were measured by a computerized image analyzer.Results The mean pulmonary artery pressure (mPAP) of normoxic group,hypoxic group and angelica solution-protected group were10.50±1.90,35.36±9.11,18.32±2.30 (mm Hg);wA% of the three groups were 52.71±5.16,82.38±8.43,64.58±9.54 (%),mPAP and wA% were significantly higher in the hypoxic group than those in the normoxic group (P<0.01) and angelica solution-protected group (P<0.01).PCNA expression of normoxic group,hypoxic group and angelica solution-protected group were 3.15±1.10,24.50±5.72,12.67±3.46 (%).The PCNA expression in the pulmonary artery was significantly higher in the hypoxic group than those in the normoxic group (P<0.01) and in the angelica solution-protected group (P<0.01).iNOS expression of normoxic group,hypoxic group and angelica solution-protected group were 2.13±1.01,17.33±3.53,37.50±7.04 (%).iNOS expression in the pulmonary artery was higher in the hypoxic group than those in normoxic group (P<0.01),and angelica significantly increased iNOS expression in comparison with the normoxic and hypoxic groups (P<0.01).Conclusion Angelica solution alleviates chronically hypoxia induced pulmonary hypertension in rats by inhibiting the espression of PCNA in pulmonary artery and up-regulating the expression of iNOS.

Adolescent ; Cardiac Catheterization ; methods ; Child, Preschool ; Echocardiography, Transesophageal ; methods ; Female ; Heart Septal Defects, Atrial ; diagnostic imaging ; therapy ; Humans ; Treatment Outcome

OBJECTIVETo observe the effect of puerarin on infarction size, fatty acids metabolism, inflammatory response and atherosclerotic plaque stability in patients with acute myocardial infarction (AMI).

METHODSSixty-one patients with AMI were randomly divided into two groups, the control group (n = 30) and the treated group (n = 31). All were treated with conventional treatment, but to the treated group, puerarin injection was given additionally by injecting 500 mg per day for 2 weeks. Before and after treatment, blood levels of free fatty acids (FFA), matrix metalloproteinases-9 (MMP-9) and C-reactive protein (CRP) were assayed, and the size of infarction was determined by Ideker QRS scoring method.

RESULTSBefore treatment, the size of infarction was positively correlated to the levels of FFA, MMP-9 and CRP (r = 0.43, 0.42 and 0.39, respectively, all P<0.01). As compared with those before treatment, after treatment, the three parameters lowered by 30%, 41% and 23%, respectively and the size of infarction significantly reduced in the treated group (P<0.01), while in the control group, no significant change was found (P>0.05).

CONCLUSIONPuerarin treatment could significantly reduce the size of infarction in patients with AMI, the mechanism is possibly related with its effects in lowering plasma levels of FFA, inhibiting inflammation and stabilizing atherosclerotic plaque.

Aged ; Biomarkers ; C-Reactive Protein ; metabolism ; Fatty Acids ; metabolism ; Female ; Humans ; Isoflavones ; therapeutic use ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; Myocardial Infarction ; drug therapy ; pathology ; Vasodilator Agents ; therapeutic use

Objective To assess the association of synovial cyclic citrullinated peptide(CCP)expression with T helper 17(Th17) cells/Regulatory T cells (Treg) imbalance,the histological and clinical features of synovitis in rheumatoid arthritis (RA).Methods CCP expression in synovial specimens from 39 patients with RA and 35 controls was detected by immunohistochemistry assay(IH) using 6×His tagged anti-CCP single chain fragment V (ScFv) antibodies,which were generated by pHEN2 phagemid recombinant antibodies display system.The frequencies of Th17/Treg cells in the peripheral blood mononuclear cells (PBMCs) were determined by flow cytometry (FCM).Th17/Treg cells associated cytokines were analyzed by enzyme-linked immunosorbent assay (ELISA).The histological scores and clinical features of synovitis were included in the study.Chi-square test and ANOVA were used for statistical analysis.Results ① The prevalences of synovial C CP expression were significantly different between RA group and the control(76.9％ and 11.4％ respectively,X2=31.9,P＜0.01).② The frequencies of Th17 cells,Th17/Treg ratio,Th17 cells associated cytokines as IL-6,IL-17a,IL-23,TNF-α,and the Treg cells associated cytokines TGF-31,the serum and synovial fluid anti-CCP antibodies in the RA patients with synovial CCP positive expression were significantly higher in RA patients with CCP positive than those with CCP negative.Disease activity score DAS28 index and histological features quantified variations of the synovial biopsy specimens (synoviocyte hyperplasia,focal aggregates of lymphocytes,and diffuse infiltrat(e)s of lymphocytes) in RA were higher in synovial CCP positive expression patients than in the negative.Conclusion Synovial CCP expression is strongly associated with the Th17/Treg imbalance and synovitis,which may play a crucial role in the pathogenesis of RA.

The expression vector of shRNA targeted to the rat angiotensin Ⅱ receptor gene wasconstructed and the efficacy of siRNAs to modulate the expression of target gene in the in vitro cul-tured mammalian cells was investigated for antihypertensive therapy in spontaneous hypertensive rat(SHR) at post-transcriptional level. The sense and antisense RNA oligonucleotides strands targe-ting angiotensin Ⅱ receptor mRNA were synthesized individually according to the sequence of therat angiotensin Ⅱ receptor. For preparation of duplexes, sense- and antisense-stranded oligonucle-otides were mixed and annealed, and the annealed duplexes were cloned into the pGenesil-1 vector..The rat glioma cells were transfected with constructed pGenesil-1-shRNA plasmid and scrambled plasmid. The cultured cells were collected at different phases. RT-PCR and Western blot were performed. The AT1 mRNA and protein levels behaved ultimately same. Compared to control after 48 h, AT1 mRNA levels were decreased to 35.5 %±3.0 %, and the levels reached their lowest point after 72 h (20.7 %±4 % of control). At 24 and 48 h, AT1 protein was reduced to 46.9 %±4.2 %and 36.98 % ± 3.7 % respectively compared to control and a maximum reduction was observed after 72 h of incubation (28. 1% ±4 % compared to controls). Plasmid-based shRNA expression systems targeted against the rat angiotensin Ⅱ receptor gene were generated successfully. The shRNAs with a 22-nt stem and a short loop were cleaved into small interfering dsRNA (siRNA) by the Dicer. The in vitro transcribed siRNA enables the effective silencing of gene expression to the target mRNA and leads to effective inhibition of translation of proteins and will be lay the foundation of application of gene silencing technology to hypertensive rats.

OBJECTIVETo study the distribution of DC-SIGN/DC-SIGNR alleles among drug user (DUs) populations with or without HIV/HCV infection in Shenzhen, and to evaluate the role of these alleles in the construction of genetic resistance to HIV or HCV and screen out the anti-HIV/HCV gene in Shenzhen.

METHODSAll 500 DU blood samples were collected from Shenzhen Detoxification Center, including 313 from injected drug users (IDUs). All samples were screened for HIV and HCV antibody by means of ELISA. The genomic DNA were extracted and amplified by PCR. The neck domain repeat regions of DC-SIGN/DC-SIGNR were sequenced directly from the PCR products to confirm the amplification for some samples and all positive PCR products were analyzed by agarose gel electrophoresis.

RESULTSOf 500 samples, 97 were found HIV positive, all of which were IDUs and HCV positive. The total positive rate of HCV among all HIV negative DU was 57.57% (232/403), and it was 63.89% (138/216) among IDUs; in comparing to the 50.26% (94/187) of DUs with other manners there showed significant difference (chi(2) = 7.61, P = 0.0058). Among HIV + DUs, there was a higher proportion of patient with the DC-SIGNR 5/6 and 5/8 (Fisher's exact, P = 0.043 and P = 0.034) with statistical significance; there was no statistically significant difference between HCV + and HCV-DUs and no significant difference between IDUs and other DUs for the DC-SIGNR polymorphism.

CONCLUSIONThe results might indicate that DC-SIGN/DC-SIGNR polymorphism might not influence the susceptibility to HCV. Genotype 5/6 might probably have a relation with HIV infection, but still need further investigation for the low frequency.

Adolescent ; Adult ; Alleles ; Cell Adhesion Molecules ; genetics ; Drug Users ; Female ; Gene Frequency ; Genotype ; HIV Infections ; genetics ; HIV-1 ; Hepacivirus ; Hepatitis C ; genetics ; Humans ; Lectins, C-Type ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Receptors, Cell Surface ; genetics ; Young Adult

OBJECTIVETo investigate the role of xenogenic (porcine) ADM as dermal substitute in scar treatment.

METHODSAfter scar excision, the wounds were covered with composite grafts of DR procine ADM and autologous thin split-thickness grafts in one stage or in two stages.

RESULTS22 out of 47 cases were treated in two-staged procedure. After the ADMs were applied to the wound, the autologous thin split-thickness grafts were implanted 7 days later. 25 cases were treated in one-staged procedure. The survival rates of composite grafts were (88.3 +/- 3.7)% for subcutaneous recipient bed and (89.7 +/- 3.4)% for deep fascia recipient bed in group with two-staged procedure, compared with (92.5 +/- 4.1)% and (93.2 +/- 5.2)%, respectively, in group with one-staged procedure. Early after grafts taken, the grafts had a pink colour and smooth surface. The patients were followed up for 90 days at most. The survived composite grafts were durable, elastic, smooth and soft with good function and appearance like normal skin. They could even be pinched up. The scar along the edge of the grafts was slightly hypertrophic.

CONCLUSIONSThe survival rate of composite graft is higher in patients with one-staged procedure. The elasticity and textural of the taken grafts are better on subcutaneous recipient bed than on deep fascia recipient bed, though the function has no difference. Xenogenic (porcine) ADM can be an optimal dermal substitute for wound coverage after scar excision.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Child ; Child, Preschool ; Cicatrix ; surgery ; Dermis ; cytology ; transplantation ; Humans ; Male ; Middle Aged ; Skin, Artificial ; Swine ; Transplantation, Heterologous ; Young Adult

BACKGROUNDNitric oxide (NO) is an important mediator in the pathophysiology of many vascular diseases. However, the definite role of NO in human abdominal aortic aneurysm (AAA) formation is unclear. The aim of this study was to investigate production of NO and expression of inducible nitric oxide synthase (iNOS), and their possible role in AAA.

METHODSA total of 28 patients with AAA, 10 healthy controls, and 8 patients with arterial occlusive disease were enrolled into this study. Standard colorimetric assay was used to examine NO concentration in plasma from patients with AAA and normal controls, and in cultured smooth muscle cells (SMCs). Expression of iNOS in aortas and cultured SMCs were detected by immunochemistry. The correlation of iNOS expression with age of the patient, size of aneurysm, and degree of inflammation was also investigated by Cochran-Mantel-Haenszel chi2 test and Kendall' Tau correlation.

RESULTSExpression of iNOS increased significantly in the wall of aneurism in the patients with AAA compared to the healthy controls (P < 0.05) and the patients with occlusive arteries (P < 0.05). iNOS protein and media NOx (nitrite + nitrate) also increased in cultured SMCs from human AAA (n = 4, P < 0.05), while plasma NOx decreased in patients with AAA (n = 25) compared to the healthy controls (n = 20). There was a positive correlation between iNOS protein and degree of inflammation in aneurismal wall (Kendall coefficient = 0.5032, P = 0.0029).

CONCLUSIONSSMCs and inflammatory cells were main cellular sources of increased iNOS in AAA, and NO may play a part in pathogenesis in AAA through inflammation.

Adult ; Aged ; Aortic Aneurysm, Abdominal ; etiology ; Apoptosis ; Female ; Humans ; Male ; Middle Aged ; Muscle, Smooth, Vascular ; pathology ; Nitric Oxide ; physiology ; Nitric Oxide Synthase Type II ; analysis ; physiology