Objective To evaluate the effect of inhibition of transforming growth factor-beta 1 (TGF-?1) expression by liposome-mediated antisense oligonucleotides on chronic nephrotoxicity induced by cyclos-porin A in rats. Methods Rats were on low salt diet and CsA was administered once every day by intraper-itoneal injection (15 mg/kg every day) for 4 weeks. Antisense oligodeoxynucleotides for TGF-?1 were introduced into the nephritic kidney by liposome-mediated gene transfer method to inhibit the overproduction of TGF-?1. Results Under the condition of low salt diet, a rat model of CsA-induced nephrotoxicity was established. The changes of level of serum creatinine and characteristic histopathology of proximal tubular injury and tubulointerstitial fibrosis were observed. In the liposome-mediated AODN rats, the expression of TGF-?1 mRNA was decreased by RT-PCR (P

Objective To carry out a taxonomic identification of a strain of claviform bacteria iso-lated from prostatic fluid of a patient who suffered from chronic prostatitis, and to approach its phylogenic and biologic position. Methods We undertaked an initial identification by phenotypic characters such as morphologecal, physiological and biochemical characteristics to ascertain its phylogeny by chemical composi-tion analysis of cell wall and 16S rRNA gene sequencing and alignment. Results A club-shaped gram posi-tive rod bacillus was isolated in pure culture state. Its biochemical reactions were not active. The diamino-acid of cell wall was meso-diaminopimelic acid (meso-DAP) and it had wall chemotype Ⅳ ( contained arabi-nose, galactose and maltose ). Sequence searches of the GenBank database revealed that this strain had a highest level of 16S rDNA sequence similarity (99.4%) to C. tuberculostearicurn strain ATCC35692 with only 8 nucleotides difference. Conclusion On the basis of phenotypic and phylngenetie analysis, it is rea-sonable to assign this strain to the species C. tuberculostearicum, and this is the first isolation of C. tubercu-lostearicum from prostatic fluid home and abroad.

Objective To identify a myeloid differentiation factor 88(MyD88)in Oncomelania hupensis,and characterize the role of MyD88 against Schistosoma japonicum infection. Methods The complete cDNA of MyD88 in O. hupensis was ob-tained by using rapid amplification of cDNA ends(RACE),and homologues sequences and conserved domains were aligned and the structure of MyD88 was predicted either. A phylogenetic tree of MyD88 was further constructed with other species. In ad-dition,the mRNA expression level of O. hupensis MyD88 before and after S. japonicum infection was investigated by real-time quantitative PCR(RT-qPCR). Results The cDNA of O. hupensis MyD88 consisted of 1406 bp open reading frame(ORF),en-coding 468 amino acid residues,which contained death domain and Toll/interlrukin-1 receptor(TIR)domain,the typical fea-tures of MyD88 family proteins. The predicted amino acid sequence of O. hupensis MyD88 shared 38%-52%identity with other mollusc. O. hupensis MyD88 was phylogenetically closeted to Biomphalaria glabrata MyD88. The O. hupensis MyD88 existed in all selected tissues and expressed highly in hemocyte,up-regulated after S. japonicum infection in all selected tissues except cephalopodium,especially higher in whole snail and hemocyte. Conclusion MyD88-dependent signaling pathway is present in O. hupensis and plays an important role in innate immune response against S. japonicum infection.

Objective To observe destruction of tibias created in ablating of the swine limbs in vivo with radiofrequency,the influence on the surrounding organization structure,and the repair process of lesions for treating bony tumor.Methods There were eight pigs included in the first part of the experiment.RFA was performed under DSA guidance at the same position of the diaphysis and the upper end of their right tibias in hind leg during anesthesia,then X-ray and CT examinations were performed at different time points after RFA,and subsequently the pigs were executed by depth anesthesia.Control specimens,specimens of the immediate time,the third day,the tenth day,the twenty-fourth day,the fifth week,the eighth week,the twelfth week after RFA were obtained at the diaphysis and the upper end of their tibias.X-ray and CT examinations of these specimens were performed.These specimens were observed both by naked eye and under microscope.The observation included the general state of the laboratory animals after RFA,the configuration of RFA lesions in the gross specimen and the corresponding histologic changes,the boundary of necrosis in RFA lesions and the corresponding boundary of the gross specimen.There were sixteen pigs in the second part of the experiment.RFA was performed at the same position of the diaphysis and the upper end of their right and left tibias in hind leg during anesthesia,and the pigs were immediately executed by depth anesthesia after RFA to get thirty-two capitulum specimens and thirty-two diaphysis specimens.The scope of necrotic tissue in RFA lesions was observed and measured.The border and scope of necrotic tissue in RFA lesions were observed.Major axis,minor axis,and necrosis volume were measured.The independent sample t test was employed to compare the volume of necrotic tissues between capitulum and diaphysis.The one sample t test was employed to analyze volume of necrotic tissue among capitulum,diaphysis and the standard data.Results No accidental death of the animals occured in the first experimental.The loss of neurological function of the hind leg did not happen.Coagulation necrosis in cancellous bone of capitulum and marrow of diaphysis lesions created in ablating was observed.On 3th to 10th day after RFA,inflammatory cells infiltrated in hemorrhage tape and then granulation tissues formed.Granulation tissues replaced necrotic bone gradually from outside to inside.Low density band was showed on images of X-ray and CT examination in the specimen of 10th day,and it extended inward.The immature bone was observed in fibrous tissue sited at the edge of lesions,which appeared as sclerosis on X-ray and CT examination.The scope of necrotic tissue in RFA lesions included coagulation necrosis and hemorrhagic zone of its periphery.The dead bone and secondary pathological fracture appeared in two of eight pigs.RFA hardly damaged compact bone in the diaphysis,but the damage of soft tissue was observed outside the compact bone.In the second experiment,the shape of lesions in cancellous bone of capitulum was ellipsoid,with long axis of(2.59±0.21) cm,minor axis of(2.15±0.10) cm,and volume of(6.34±0.27) cm3.The shape of lesions in the diaphysis of tibias was ellipsoid,with long axis of(4.53±0.20) cm,minor axis of(1.71 ±0.22) cm,and volume of(7.14±0.36) cm3.There was significantly difference of necrosis volume between the capitulum and diaphysis lesions,(t=2.011,P＜0.05).The virtual necrosis volume of the capitulum and diaphysis lesions was significantly different from the presumed volume with RFA instrument(t=-613.371,-295.878,P＜0.01).Conclusion Cancellous bone of the capitulum can be damaged by RFA,but the compact bone of the diaphysis can hardly be damaged.The intact compact bone can be helpful to protect the surrounding soft tissues.X-ray and CT examination can be used in the evaluation of curative effect of RFA for treating bone tumors.Pathological fracture of the diaphysis may appear after RFA.

Objective To study the influence of hemodiafiltration on high-sensitivity C-reactive protein (hs-CRP),serum parathyroid hormone (iPTH) and cardiac function in uremic patients undergoing maintenance hemodialysis (MHD).Methods Forty patients with uremia were divided into hemodiafiltration (HDF) group (n =20) and hemodialysis(HD) group(n =20) by random numbers.Serum hs-CRP and iPTH were measured before treatment and at 6 months after treatment.Their left ventricular diastolic diameter (LVEDD),left ventricular end-systolic dimension (LVESD),interventricular septal thickness (IVST) and left ventricular ejection fraction(LVEF) were measured by echocardiogram.Ten healthy volunteers were recruited as the normal controls.Results After treatment,serum hs-CRP and iPTH decreased significantly in the HDF group (hs-CRP:(3.32±1.t4) mg/L vs.(7.84 ±2.21) mg/L,t =2.072,P =0.046;iPTH:(155.36 ±81.33) ng/L vs.(190.25 ±82.31) ng/L,t =2.121,P =0.023).No significant difference of serum hs-CRP and iPTH was observed in the HD group before and after treatment (hs-CRP:(7.91 ±2.11) mg/L vs.(7.68 ± 1.86) mg/L,t =1.731,P =0.111 ; iPTH:(177.43 ± 85.43) ng/L vs.(184.35 ± 87.21) ng/L,t =1.331,P =0.210).Color Doppler indices of cardiac structure of the HDF group significantly improved after treatment compared with before treatment (LVEDd:(55.3 ±3.7) mm vs.(58.8 ±4.3) mm,t =2.345,P =0.015 ;LVESd:(33.5 ±2.6) mm vs.(36.4±4.3) mm,t=2.178,P=0.046;IVST:(10.2±1.7) mmvs.(13.1 ±1.8) mm,t=2.630,P=0.012;LVEF:(55.3 ±2.6)％ vs.(42.5 ±3.3)％,t =2.860,P =0.010;LVMI:(132.5 ±9.1)g/m2 vs.(137.4 ± 8.7) g/m2,t =2.871,P =0.009).After treatment,cardiac structure of the HDF group was significantly better than that of the HD group (LVEDd:(55.3 ± 3.7) mm vs.(59.1 ± 4.2) mm,LVESd:(33.5 ±2.6) mm vs.(36.1 ±3.4) mm,IVST:(10.2±1.7) vs.(12.4 ±1.3) mm,LVEF:(55.3 ±2.6)％vs.(43.4±2.7)％,LVMI:(132.5 ±9.1) g/m2 vs.(139.4 ±8.9) g/m2;P ＜0.05).During the six month treatment,the incidence rate of cardiovascular events of the HDF group was significantly less than the HD group (36.39％(524/1440) vs.(72.08(1038/1440),x2 =21.583,P＜0.01).Conclusion HDF may efficiently remove iPTH,improve cardiac function of uremic patients,and help them recover from micro-inflammatory state.

Objectives To observe the expressions ofα-smooth muscle actin (a-SMA) and type III collagen (Col-III) of tubuloin-terstitial ifbrosis(TIF) induced by unilateral ureteral obstruction (UUO) in rat and the intervention effect of supplemental hydrogen sul-ifde (H2S). Methods Ninety-six male Sprague-Dawley rats were randomly divided into 4 groups, sham-operated group, UUO model group, NaHS low-dose group and high-dose group. TIF rat model was established via UUO. After UUO operation, low-dose and high-dose group were intraperitoneally injected twice a day with 1.4μmol/kg and 7.0μmol/kg NaHS, respectively. Sham-operated group and UUO model group were given an equivalent volume of normal saline. Eight rats in each group were killed randomly at 7, 14 and 21 days after UUO operation. The concentration of plasma H2S was detected using deproteinization. Renal tubulointerstitial damage was evaluated with routine Hematoxylin and Eosin staining and Masson staining under microscope. The expressions ofα-SMA, Col-III were measured with immunohistochemistry. Results Compared with sham-operated group, renal tubulointerstitial injury was severer in UUO model group and was alleviated after intervention of NaHS. There was signiifcant difference in tubulointerstitial injury among all groups (P<0.01), but no difference was found between high-dose and low-dose group. Exogenous H2S supplement could down-regu-late the expressions ofα-SMA and Col-III in renal tissues (P<0.05). There was no difference between high-dose and low-dose group in the expressions ofα-SMA and Col-III (P>0.05). Conclusions TIF induced by UUO is associated with decreased level of endogenous H2S. H2S supplementation can ameliorate the development of UUO-associated TIF in part through down-regulating the expressions ofα-SMA and Col-III in renal tissues. However, a dose dependent manner between the two doses of exogenous H2S supplementation was not observed.

Ablation Techniques ; adverse effects ; Animals ; Bone and Bones ; pathology ; Disease Models, Animal ; Extremities ; Swine

Objective To investigate the effects of 5-Aza-CdR (methylation transferase inhibitor) on gene expression levels of interleukin-1β (IL-1β),transforming growth factor-β (TGF-β) and nitric oxide synthase (NOS) in chondrocytes.Methods Chondrocytes from patients with osteoarthritis (OA) (n=22),rheumatoid arthritis (RA) (n=3) or trauma without rheumatic diseases (n=10) were collected and cultured.The mRNA expression levels of IL-1β,TGF-β and NOS were detected by real-time polymerase chain reaction (RT-PCR).Chondrocytes in trauma group were treated with 5-Aza-CdR(10μmol/L) for 72 h,and the mRNA and protein expression levels of IL-1β,TGF-β and NOS were detected by RT-PCR and ELISA,respectively.Results The mRNA expression levels of IL-1β,TGF-β and NOS were increased in OA and RA group as compared with trauma group (P＜0.05),while they had no differences between OA and RA groups.After treated with 5-Aza-CdR,the mRNA and protein expression levels of IL-1β,TGF-β and NOS in chondrocytes rised in trauma group as compared with pretreatment (all P＜0.05).Conclusions The mRNA expression levels of IL-1β,TGF-β and NOS in chondroytes are higher in OA and RA patients.5-Aza-CdR could increase the mRNA and protein expression levels of IL-1β,TGF-β and NOS by inducing relative gene methylation,which suggests demethylation might play a role in OA pathogenesis by influncing the inflammatory signal pathway or cell apoptosis.

Objective Tubulointerstitial fibrosis(TIF) is the most important marker reflecting the degree of renal function decline and prognosis and hydrogen sulfide ( H2 S) is crucial in maintaining normal renal function and many diseases of renal injury. The aim of the article was to investigate the effects of exogenous H2 S on the expressions of angiotensinⅡ ( AngⅡ) , proliferating cell nuclear antigen (PCNA) and transforming growth factor beta-1 (TGF-β1) in rats with unilateral ureteral obstruction (UUO). Methods TIF rat model was built with UUO. Ninety-six SD rats were randomly divided into four groups:sham operation group, modelgroup, UUO+low-dose NaHS treatment group ( low dose group) and UUO+high-dose NaHS treatment group ( high dose group) ( n=24, respectively) . Rats in model group were treated with left-side ureteral obstruction and ureteral separation without obstruction was done in sham operation group. UUO rats in two treatment groups were injected intraperitoneally with two different doses of sodium hydrosulfide (NaHS, donor of endogenous H2 S), respectively. HE and Massonstaining and immunohistochemical staining were performed at the 7 d, 14 d and 21 d, respectively. Results In sham operation group, the expressions of AngⅡ, PCNA, and TGF-β1 were found in microamount in tubulointerstitium at each time points. Compared with sham operation group, the expressions of AngⅡ, PCNA and TGF-β1 in model group increased( P<0.01) . While in comparison to model group, the expressions of AngⅡ, PCNA and TGF-β1 decreased in low dose group and high dose group, but no significant differ-ence was found between low dose group and high dose group. Conclusion Exogenous H2 S supplementation can attenuate TIF partly via downregulating the expressions of AngⅡ, PCNA and TGF-β1 .