Objective To investigate of human esophageal squamous carcinoma cells(Eca-109) in which cyclooxygenase (COX)-2 gene was knocked down by adenovirus delivered siRNA.Methods Based on the plasmid pSUPER cloned with RNA polymerase Ⅲ-dependent promoter HI,the interfering plasmid psiRNA/COX-2 targeting human COX-2 mRNA was constructed,The siRNA/COX-2 fragment was derived from psiRNA/COX-2 digested by Not I and Xho 1.and was cloned into the shuttle plasmid pAdTrack.Then pAdTrack/siRNA/COX-2 was obtained and co transfected into the E.coli strain BJ5183 with the bone plasmid pAdEasy-1,the recombinant adenovirus Ad/siRNA/COX-2 was generated by homologous recombination.Having been packaged and amplified in cells 293,Ad/siRNA/COX-2 was transfected into Eca-109 cells.The PGE2 concentration in the cells culture supernatant was determined by ELISA,and the level of COX-2 mRNA in the cells was tested by real time PCR.Moreover,cell cycle and apoptosis were determined by flow cytometry,and cells growth curve was protracted.Results The recombinant adenovirus Ad/siRNA/COX-2 was successfully constructed.Ninty six hours after Ad/ siRNA/COX 2 transfecting into Eca 109 cells,COX-2 mRNA was reduced by 71.7%,and PGE2 concentration in the cells culture supernatant was decreased by 62.0%.Correspondingly,the growth of cells slowed down.At the same time,the cells in G0-G1 phase was increased by 32.24%,and those in S phase and G2-M phase were reduced by 16.38% and 15.86%.respectively.And cells apoptosis index was increased by 9.19%.Conclusion The adenovirus based-RNAi was capable of knocking down remarkably COX-2 of human esopbageal carcinoma cells,which lead to growth of cells slowing down.

Objective To study the clinical features of follicular occlusion triad, and whether it is a hereditary disease. Methods Based on the clinical examination of a case who developed squamous cell carcinoma secondary to follicular occlusion triad, the pedigree of the patient was surveyed and analyzed. Results There were a total of thirteen patients in this pedigree, the age of onset was about 20 years old. The clinical features and laboratory examination of the proband was consistent with follicular occlusion triad. Conclusions Hereditary factor is important in the pathogenesis in follicular occlusion triad,and the disease maybe an autosomal dominant inherited disease.

Objective To measure the parameters of anterior bow of the femoral medullary cavity by means of 3D spiral CT reconstruction and to provide the anatomical basis for the design of the femoral component prosthesis.Methods 105 healthy Chinese left femurs were scanned in standard neutral position by 3D spiral CT.Parameters of the anterior bow of the femoral medullary were measured by medical imagine software.Results The parameter values of the male femur were statistically bigger than those of the female,including the length of the femoral medullary and the distance from the point of maximum curvature to plane above lesser trochanter (P＜0.001).No significant differences between male and female in the position index of the point of maximum curvature were observed (P＞0.05).The relative curvature of the female femoral anterior arch was statistically bigger than that of the male (P＜0.001).The femoral curvature subtense and the length of the femoral medullary showed no significant correlation (r=0.106,P=0.281).Conclusion There are certain anatomic differences in the anterior bow of the femoral medullary between male and female.Appropriate length of femoral component should be considered in total hip replacement operation.And it is necessary to design the femoral components fitting the features of the femurs according gender difference.

Two patients with neonatal diabetes tested as V59A and V59M mutations were chosen for the study. Clinical data were analyzed retrospectively. The results showed that the patient with V59A mutation was characteristic of spasm and hyperglycemia at the age of three month, and treated with insulin for a long time as unresponsive to the glibenclamide at the beginning. Myasthenia and delay of development were observed during the follow-up. At the age of two years, glibenclamide was tried for the second time with a high dose and fairly-controlled glucose level. The patient with V59M mutation was diagnosed with diarrhea, hyperglycemia, and ketosis at the age of two month, and was responsive to glibenclamide at a relatively low dose with well-controlled glucose level. These results suggest that KCNJ11 V59M mutation would show some milder clinical manifestations and better glibenclamide efficacy as compared with V59A mutation.

Objective To evaluate the diagnostic value of velocity parameters of Tardus-Parvus for the detection of renal artery stenosis ( RAS) ( diameter reduction ≥50%) and to determine the useful cutoffs for these parameters .Methods A study group was composed of 221 renal arteries that were detected Tardus-Parvus by color Doppler flow imaging and were referred to abdomino-aorto-re-nal arteriography afterwards .Five Doppler parameters including the peak systolic velocity ( PSV ) , end-diastolic minimum velocity (EDV), resistance index (RI), acceleration time (AT), and accelerated velocity (AC) from each location including renal aorta ( MRA) , segmental artery ( SRA) , interlobar artery ( IRA) , and arcuate artery ( ARA) were archived and compared among the differ-ent groups .Renal artery angiography showed arterial canon reduced 50% or higher RAS .Arteries were considered stenosed on renal arteriography if there was a diameter reduction of greater than 50%.Statistical analysis to determine the best parameter for predicting a RAS was performed with the receiver operating characteristic ( ROC) curves.The sensitivity, specificity, and negative and positive predicting values at various cutoffs were calculated .Results Renal artery stenosis degree was less than 50% in 16 cases, 50%~99%in 197 cases, completely occluded in 2 cases, and no stenosis in 6 case by renal arteriography .For RAS with inner diameter re-duction of 50%or more, the ROC curve analysis showed renal artery flow velocity had a good sensitivity and specificity , 50%~99%of the RAS had optimal threshold value in PSV 20 cm/s, EDV 10 cm/s, RI 0.50, AT 0.09 s, and AC 1.5 m/s2.Conclusions Tardus-Parvus had high specificity and sensitivity for diagnosis of stenosis rate in more than 50% of the RAS, low sensitivity for the narrow degree in 0~49%, and no clinical value for the renal artery occlusion .

Objective To identify two suspected Legionella pneumophila (L.pneumophila) strains isolated from environmental water and air conditioning cooling water systems in Guangzhou city.Methods The two strains were identified by their cultural characteristics,biochemical test,Legionella-specific primer PCR identification,PCR-enzymatic digestion analysis,16S rRNA,mip and rpoB gene sequencing analysis.Results The two suspected L.pneumophila isolates were identified as gram-negative bacillus appeared as white colonies on BCYEα-agar after incubation for 48 hours at 36℃.Both isolates were positive for oxidase,gelatinase and hydrolysis of hippurate,and negative for urease activity and nitrate reduction.Their phenotypic characteristics were similar to those of L.pneumophila strains.Results of PCR identification by using Legionella-specific primer were positive.Enzymatic digestion analysis showed that the 226 bp PCR products of two isolates were not digested by Taa Ⅰ.The two strains were classified as Legionella busanensis as indicated by gene sequencing analysis of 16S rRNA,mip and rpoB gene.Conclusion Two L.busanensis strains were first isolated from environmental and air conditioning cooling water systems in China.Due to their biochemical characteristics,L.busanensis strains were commonly misidentified as L.pneumophila,but could be effectively identified by PCR-enzymatic digestion analysis and multiple genes identification.

Objective To discuss the technical points, safety and clinical effectiveness of DSA-guided percutaneous acetabuloplasty (PA) for the treatment of acetabular metastases. Methods Fifteen patients, including 6 males and 9 females, with acetabular malignant metastases were enrolled in this study.A total of 19 lesions were detected. The lesions were 13 - 25 mm in size (mean 19 ± 4 mm), the dose of PMMA used for per lesion was 5 - 13 ml (mean 8.2 ± 2.3 ml). All patients complained of greater or less degree of pain in their hips, 8 patients had to use walking stick, 4 patients showed limping although they could walk independently and three patients could not walk alone. DSA-guided percutaneous acetabuloplasty was performed in all patients and follow-up after the procedure was conducted for 1 - 12 months. Visual analogue score (VAS), walking state score and analgesic dosage taken by the patients were used for the evaluation of the clinical effectiveness. The complications were analyzed. All the data obtained were statistically analyzed with paired samples t test and analysis of variance by using SPSS12.0 statistical software.Results The procedure was technically successful in all patients. Pain rating evaluated by the VAS decreased from a mean of 7.8 before surgery to a mean of 4.2 in 24 hours after surgery (P < 0.01 ), which further decreased to 2.5 in one month (P < 0.01 ), while walking state score increased from a mean of 1.5before surgery to a mean of 2.5 in 24 hours after surgery (P < 0.01 ). The analgesic dosage taken by the patient was reduced in 14 patients and remained the same in one patient. The bone cement leakage into paraacetabular soft tissues occurred in three cases and peripheral vascular exudation of acetabulum was observed in three cases with no obvious clinical symptoms. Conclusion As a safe, reliable and minimally-invasive technique, DSA-guided percutaneous acetabuloplasty has excellent anti-pain effect in treating acetabular metastases. This therapy can markedlyimprove the patient's walking ability and the quality of life.

Background and purpose: Hepatocellular carcinoma (HCC) is a hypervascular tumor associated with a poor prognosis and lack of effective treatments. Consequently, identifying novel therapeutic strategies are urgently needed. We have previously shown that the kringle 1 domain of human hepatocyte growth factor (HGFK1) is a more effective anti-angiogenesis molecule than angiostatin. In this study, we observed the effects and mechanisms of HGFK1 gene on the HCC. Methods: A recombinant adeno-associated vires carrying the HGFK1 gene (rAAV-HGFK1) was constructed.HCC of rat was induced by McA-RH7777. rAAV-HGFK1 was used to treat the rat, median survival time and metastasis rate were observed. Results: Ten days after tumor cell inoculation, surgery were performed to confirm the tumor formation, PBS, rAAV-EGFP or rAAV-HGFK1 was injected directly into the tumor nodule followed by portal vein injection. Results from our study demonstrated that rAAV-HGFK1 treatment significantly prolonged the median survival time of the HCC bearing rats from 30 days (PBS and rAAV-EGFP groups) to 49 days (rAAV-HGFK1 group). More importantly rAAV-HGFK1 inhibited tumor growth and completely prevented liver, lung and peritoneal metastasis. In the controlled PBS and AAV-EGFP group, liver and peritoneal metastasis rate were both 100%, and lung metastasis rate was 100% and 83%, respectively. While there was no metastasis found in treatment group, with only 33% of ascites happened. This was most possibly due to the primary tumor in liver but not due to the metastasis. Moreover, at a higher magnification (1000×), it was clear that the HGFK1 protein was expressed mainly in the cytoplasma of liver cells. In parallel, IHC staining of CD31 also demonstrated a significantly lower level of microvessel density (MVD) (6.21±1.6) in the liver tumor of the AAV-HGFK1 treatment group, as compared to the two control PBS and AAV-EGFP groups (25.1±2.1 and 26.8±2.5, respectively, P<0.01). HE staining showed that AAV-HGFK1 treatment induced large areas of necrosis in the tumor tissues, while minimal areas of necrosis were observed in the tumor tissue in the control groups. In addition, no toxicity appeared when high dosage (4.8× 1012 vg/rat) of rAAV-HGFK1 was administered in rats. Conclusion: Results from this study demonstrated that HGFK1 inhibited the growth and metastasis of HCC and prolonged the survival time of animals with HCC through anti-angiogenesis effects. No obvious toxicity was observed. It might be the novel promising treatment for HCC and other cancers.

Objective To explore the clinical application value of proton magnetic resonance spectroscopy (1H-MRS) in the metabolism of the hippocampus in non-demented Parkinson's disease(PD) patients. Methods 1H-MRS localized to both hippocampus,was carried out in 36 non-demented PD (14 patients with Hoehn-Yahr stage Ⅰ,8 patients with Hoehn-Yahr stage Ⅱ,8 patients with Hoehn-Yahr stage Ⅲ ,and 6 patients with Hoehn-Yahr stage Ⅳ) and 20 age-matched healthy control subjects. The metabolite ratios of N-acetylasparatate (NAA)/creatine (Cr),and choline-containing compounds (Cho)/Cr in PD patients were compared with those in the age-matched healthy control subjects. Results NAA/Cr ratios were reduced significantly and Cho/Cr ratios were increased sig-nificantly in both hippocampus of patients with Hoehn-Yahr stage Ⅳ compared to those in the age-matched healthy control subjects (P<0.05). However,no significant differences in NAA/Cr and Cho/Cr ratios were found between patients with Hoehn-Yahr stage Ⅰ-Ⅲ and the age-matched healthy control subjects (P>0.05). Conclusion 1H-MRS can provide the metabolic information ,and reflect the pathological changes of nerve cell in the hippocampus of non-demented PD patients with Hoehn-Yahr stage Ⅳ. It is helpful to predict the risk of dementia in PD.

Objective To investigate whether fasting obestatin level is different in patients with impaired glucose tolerance or type 2 diabetes, and to explore the association between obestatin and lipid metabolism. Methods Eighty-four subjects without known diabetes were divided into three groups: normal glucose tolerance(NGT), impaired glucose tolerance (IGT) and type 2 diabetes (DM) Plasma obestatin levels were measured with a radioimmunoassay. The relationship between fasting obestatin levels and metabolic parameters was also analyzed. Results Fasting obestatin levels were lower in DM group [(2.82±0.78)ng/ml] and IGT group [(3.25±0.29)ng/ml] than in NGT group[(3.55±0.57) ng/ml, P＜0.01]. Triglycerides and low density lipoprotein cholesterol levels gradually increased among the three groups (P＜0.05). Multiple linear regression analysis revealed fasting obestatin level was independently associated with waist-to-hip ratio, triglyeride and low density lipoprotein cholesterol. The regression equation was obestatin=6.953-3.412×W/H-0.175×TG-0.123×LDL-C. Conclusions The decreased obestatin may be associated with IGR and T2DM, and obestatin level may be associated with lipid metabolism.