To analyze the rDNA sequence of the Anisakid nematodes with zoonotic potential from Taiwan straits, Anisakid nematodes were obtained from the gut of marine fish and identified chiefly based on the morphological characteristics. The genomic DNA of nematodes were extracted, and the sequence of rDNA ITS were amplified by PCR using universal primers, then cloned and sequenced bidirectionally. Sequences analysis was conducted by blasting database and software DNAMAN. Results of the blasting database showed that no match could be demonstrated in 3 of 6 rDNA sequences obtained from 6 different nematode species;the new sequences were submitted to the database and the accession numbers in GenBank were obtained. The relationship between adult Contracaecum muraenesoxi and its putative third stage larva was validated by molecular evidences. Sequences relationship based on the inner similarity of the internal transcribed spacer sequences was constructed. It was found that the phylogenetic informative loci were mainly gathered in the second internal transcribed spacer(ITS2) for these anisakid nematodes because of the ITS2 has higher transition ratio than that of the ITS1. It is evident that the sequence in this region could provide valuable information for the molecular distinction of anisakid nematodes with zoonotic potential from Taiwan straits.

Armillariella mellea polysaccharide ( AP ) was isolated from solid fermented Armillariella mellea. AP ( 100 mg/kg?d, ig ? 5 d ) increased the production of serum hemolysin in normal mice as well as in im-munodepressed mice induced by cyclophosphamide. At the dose of 50 mg/kg?d ig?5d, AP also caused a significant increase of spleen plaque forming cells ( PFC ) in normal mice. AP ( 10, 50?g/ml ) markedly enhanced Con A-induced lymphocyte proliferation of mouse spleen cells in vitro, but it had no potentiating effect on delayed cutaneous hypersensitivity ( DCH ) to 2,4-dinitrochlorobenzene ( DNCB ) in normal mice. AP ( 100 mg/kg/d, sc? 7d) increased both clearance rate of iv charcoal particles and phagocytic activity of ma-crophages of abdominal cavity in normal mice.

Objective To introduce a better and quicker technique of transperitoneal laparoscopic ureterolithotomy. Methods 42 cases of ureterolithiasis (23 males and 19 females;mean age of 42.4 years,range from 16 to 63 years) were included.Of them 26 cases had calculi on the left side and 16 cases,on the right side.All the cases underwent laparoscopy through 3-port technique. Results The operations were successful in all of 42 cases.The mean operative time was 43 min (range,33 to 56 min).Blood loss was 20 to 50 ml.The postoperative hospital stay was 3 to 5 days.None experienced urinary leakage.During the follow-up of 12～18 months no ureteral stricture was found.The renal functions were improved. Conclusions Our experience indicates that appropriate trocar location,exactly checking of calculi,good D-J tube drainage and skillful intra-abdomenal suturing are the keys to perform laparoscopic ureterolithotomy quickly.

Objective To describe the new technique of transperitoneal laparoscopic dismembered pyeloplasty and to evaluate its clinical effect. Methods Of the 18 cases of ureteropelvic junction (UPJ) obstruction who underwent transperitoneal laparoscopic dismembered pyeloplasty,8 cases had severe hydronephrosis;7,intermediate;3,mild. Results Operations were all successful in 18 cases with the operating time being 100 to 210 min (mean,142 min) and the blood loss,40 to 200 ml.All the pores of puncture healed at phase 1;mean hospitalization was 7.4 d.Only 1 case had postoperative urine leakage.Follow-up of 3 to 12 months showed there was no stricture at UPJ and hydronephrosis was remitted (intermediate hydrops in 4 cases,mild in 7 and resolution in 7).The renal functions were improved. Conclusions Laparoscopic dismembered pyeloplasty is an effective and minimally invasive approach and can be used as an alternative to open surgery for UPJ obstruction.

0.05).The results of Western blotting and immunohistochemical method showed that HO-1 protein expression increased in DOX group and DOX-IR group,and was negative in normal group.More distruction of ultrastructural changes of hepatic cells in IR group was found than in DOX-IR group by transmission electron microscope.Conclusion Doxorubicin pretreatment could protect the liver from ischemia-reperfusion injury,which may be related to HO-1 expression induced by doxorubicin,low dosage of doxorubicin does little harm to rat liver.

Objective To evaluate the advantages of cytomegalovirus (CMV) real-time PCR, to monitor CMV infection in a population of transplantation recipients in comparison with the qualitative PCR method.Methods 150 plasma samples and leukocytes samples collected from 59 bone marrow transplant recipients and 9 liver transplant recipients were tested by qualitative CMV PCR assay in parallel; Real-time CMV PCR using Roche Light Cycler was performed with 150 plasma samples and 54 control plasma samples.Results Using real-time PCR as the reference standard, the sensitivity and specificity of the qualitative CMV PCR assay with leukocytes samples were 62.5% and 95.5%, with plasma samples were 57.5% and 99.1% respectively. The recipients with the level of CMV DNA over 5?10~3 copies/ml by real-time PCR had higher percentage of developing CMV disease than those below 10~3 copies/ml.Conclusion The quantitative detection of CMV DNA by real-time PCR with plasma is a rapid, specific and sensitive method to monitor CMV infection in patients after transplantation and to guide antivirus therapy.

ObjectiveTo investigate the expression and biological significance of PIM1 in the breast cancer,atypical hyperplasia breast,and normal breast tissues.MethodsThe protein expression levels of PIM1 in the breast cancer,atypical hyperplasia breast,and normal breast tissues were assayed by immunohistochemistry and Western blot.ResultsThe expression rates of PIM1 protein were 78.75％ (63/80)in breast cancers,42.00％ (21/50) in atypical hyperplasia tissues,and 23.33 ％ (7/30) in normal breast tissues,respectively.A significant correlation was found between PIM1 expression and the clinical stage and lymphonodus metastasis.However,no significant correlation was found between PIM1 expression and patients'age and tumor sizes.The level of PIM1 protein in breast cancers was obviously higher than that in non - cancerous tissues.ConclusionsThe positive rate of PIM1 in breast cancers is significantly higher than that in atypical hyperplasia breast tissues and in normal breast tissues.PIM1 may be an early molecularevent in mammary tumorigenesis,and its overexpression may significantly relate to the malignant progression.It would be a new parameter for the early diagnosis and a biomarker for breast cancers.It is feasible to utilize paraffin specimens for index test with advantages of convenience,easy availability,and low expense.

Objective To investigate the blood pressure circadian rhythm in patients with IgA nephropathy by ambulatory blood pressure monitoring and explore its role in the disease progression. Methods A cross sectional study was carried out.Blood pressure rhythm was studied by ambulatory 24-hour monitoring with a portable oscillometric recorder in selected patients with primary IgA nephropathy.The term dipper was described as blood pressure during night dropped at least 10％ below daytime blood pressure.The term non-dipper referred to those in whom the nocturnal decline in blood pressure was less than 10％.Clinicopathological indices between dipper and non-dipper groups were compared. Results Ninety-three patients completed ambulatory blood pressure monitoring among whom 68 (73％) patients were non-dipper.The frequency of non-dipper was 70％,70％ and 81％ in the patients at chronic kidney disease stage 1,2 and 3 or more.The frequency did not differ among these three group patients (P=-0.587).77％ of patients with hypertension and 69％ of patients with normotension were non-dipper (P=0.373).The disappearance of blood pressure circadian rhythm in IgA nephropathy was not influenced by age,gender,blood pressure,proteinuria,renal function and renal pathology lesions.Among the patients who were followed up regularly for more than 12 months (n=54),patients in the dipper group had a trend of slower eGFR decline rate than those in non-dipper group albeit the difference was not significant (P=0.329).Subgroup analysis revealed that in patients with hypertension and non-dipper (n=29),the eGFR decline rate was much faster than that in dipper group[(-6.79±11.58 )vs (-0.34±1.74) ml ·min-1 ·(1.73 m2)-1·year-1,P=0.019]. Conclusions Most patients with IgA nephropathy present disappearance of blood pressure circadian rhythm,even among those at an early stage or without hypertension.The loss of blood pressure rhythm may be associated with a rapid renal function decline rate in those with hypertension.

Objective To construct a eukaryotic expression vector of decorin(DCN),and observe its expression in CHO cells,in order to provide a basis for further study on the anti-tumor effect of DCN. Methods DCN cDNA was amplified by PCR.The human full-length DCN cDNA ligated into pBluescript was used as template.The fragment was ligated to the expression vector pCDNA3 previously digested with XbaⅠ and EcoR Ⅰ.The ligation mixture was transformed into competent E.coli JM109 cells.Transformants containing inserts were confirmed by restrictive digestion and DNA sequencing.The expression vector was transfected into CHO cells using lipofectamine,and transfected cells were cultivated in DMEM containing G418 (800 mg?L-1) for about 2 months.Immunohistochemistry method was used to detect the expression of DCN protein in stably transfected cells.Results The PCR product was about 1 000 bp.The recombinant expression vector was identified by restrictive digestion and DNA sequencing. DCN protein was detectable in stablely transfected cells.Conclusion The recombinant eukaryotic expression vector pCDNA-DEC is constructed successfully and stablely transfected CHO cells are established.

Objective:To study influence of renal sympathetic denervation (RDN)on cardiac function of dogs with heart failure (HF).Methods:A total of 40 dogs were randomly and equally divided into RDN group [received bilat- eral renal artery radiofrequency ablation (RFA)]and model group (only received femoral puncture).Pacemaker was implanted in every dog,and dog HF model was established using rapid right ventricular pacing.Cardiac and re-nal function indexes,BNP and sympathetic activity index levels were observed and compared between two groups be- fore RFA/sham operation,instant and four weeks after model establishment.Results:After operation four weeks, compared with model group,there were significant reductions in levels of epinephrine (E)[(362.69±42.54)ng/ml vs.(290.36±42.32)ng/ml],renin (R)[(305.46± 39.68)ng/ml vs.(230.04±32.80)ng/ml],aldosterone (AD)[(408.00±38.56)ng/ml vs.(246.00± 48.37)ng/ml],angiotensin Ⅱ (ATⅡ)[(280.00±48.08)pg/ml vs.(172.00±25.04)pg/ml]and norepinephrine (NE)[(425.65±50.54)ng/ml vs.(316.76±46.29)ng/ml]in RDN group (P<0.05 all);there were significant reductions in HR,respiratory rate (RR)and BNP level in RDN group,P<0.05 all;there were significant rise in SBP,LVEF,CO,CI,left ventricular pressure maximal rising rate (+dp/dtmax),left ventricular pressure maximal dropping rate (-dp/dtmax)and left ventricular end-systolic pressure (LVESP),and significant reductions in left ventricular end-systolic dimension (LVESd),left ventricular end-diastolic dimension (LVEDd)and left ventricular end-diastolic pressure (LVEDP)in RDN group,P<0.05 all.Conclusion:RDN can decrease renal sympathetic activity,improve heart function,inhibit myocardial remode- ling,its therapeutic effect is significant