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.

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

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.

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 .

Objective To explore how does chymase affect diabetic cardiomyopathy by investigating the gene expression of chymase and the relationship between the gene expression of chymase and the angiotensin Ⅱ in the cardiomyopathy of streptozotocin-induced diabetic hamasters. Methods Diabetic hamsters were induced by intraperitoneal injection of STZ 40 mg/kg once a day for 3 days. After stabilization of diabetic state for 18 weeks, the myocardial ultrastructure was observed with electron microscope and pathologic changes were observed by light microscopy. Immunohistochemistry was used to measure the level of expression of type Ⅰ and type Ⅲ collagen in diabetic and normal hamster hearts. Level of blood glucose, lipoprotein was determined using biocehemical methods. Apoptosis of cardiomyocyte was measured using TUNEL methods. Radioimmunuoassay method was used to determine the level of angiotensin Ⅱ. RT-PCR was used to determine chymase gene expression (corrected by?-actin). ResultsComparing with the control group, levels of serum glucose, TG, TC, LDL in DM group were much higher. Concentrations of collagen Ⅰ, Ⅲ and angiotensin Ⅱ [(95.8?16.0)?g/kg tissue vs (51.1?20.8)?g/kg tissue] in myocardial tissue in DM group were much higher than those in control group. RT-PCR result showed: Comparing with the control group, the mRNA expression of chymase in DM group was promoted significantly (0.810?0.026 vs 0.490?0.087). Conclusion In diabetic hamsters, the gene expression of chymase were much higher than thatin the control group, accompanying higher level of Angiotensin Ⅱ, higher levels of expression of collagen Ⅰ and Ⅲ. This result suggests that chymase plays an important role in the diabetic cardiomyopathy by promoting the activity of chymase.

OBJECTIVE To find out the status about detection and drug resistance of Ureaplasma urealyticum(Uu) and Mycoplasma hominis(Mh) of urogenital tract infection in Shaoxing area of Zhejiang Province.METHODS The detections and drug sensitive tests of Uu and Mh adopting the reagent boxes produced by bioMerieux Company in France were made.RESULTS In 2678 samples of suspicious urogenital tract infection patients,980 samples cultured of mycoplasma were positive,and the positive rate was 36.6%.Among these cases,763 samples(28.5%) were Uu positive;116 samples(4.3%) were Mh positive.In the drug sensitive test,Uu and Mh were most sensitive to PRI,JOS,DOX and TET.To CFLX,OFL,ERY,CM and AM,they had the highest level of drug resistance.CONCLUSIONS The clinical therapy of mycoplasma infection ought to choose the appropriate antibiotics based on the results of the drug sensitive test of the laboratory.

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 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 To improve clinicians'understanding of nephrotic syndrome and myasthenia gravis as adverse events of penicillamine(D-PA).Methods Two patients were reported to develope adverse reaction after taking D-PA.The related literature were reviewed.Result The first case was a systemic sclero- sis(SSc)patient treated with D-PA who developed nephrotic syndrome and blepharoptosis.The second case was a rheumatoid arthritis patient treated with D-PA who developed dysphagia and drinking bucking.The clinical symptoms were improved after disconting D-PA.Their conditions were stable up to now.Conclusion D-PA is used widely for various of autoimmune diseases.Clinicians should pay attention to these special and rare adverse effects.