Objective To dynamically observe photodamage of subcellular sites by use of laser scanning confocal microscopy (LSCM). Methods The samples were divided into four groups. Murine lung endothelial cells were subcultured and incubated with HMME for 24 hours. Then the cells were stained with rhodamine-123 for demonstration of mitochondria. LSCM was applied and organelle-cell fluorescence intensity ratio analysis was adopted to study the intracellular distribution of HMME. Then dynamic fluorescence images sequence of rhodamine-123 was collected. Results Rhodamine-123′s fluorescence images of cell sample with HMME was changed gradually during irradiation: the typical characteristic of mitochondria disappeared gradually with decreasing fluorescence intensity. The fluorescence of rhodamine-123 was diffused and distributed in nuclear, while rhodamine-123′s fluorescence images of cell sample without HMME was not changed. Conclusion Mitochondria and nucleus are photodamage sites by HMME-PDT; LSCM can be applied in dynamic observation of photodamage of subcelluar sites.

AIM: To examine Nucleostemin (NS) expression in tumor cells, and observe the effect of NS specific RNA interference on the cell proliferation in Hela cells. METHODS: Total RNA was extracted from 6 kinds of cultured tumor lines, the NS expression level was measured by RT-PCR and Northern blot. An NS-specific siRNA expression vector was constructed to transfect HeLa cell (NS-siRNA-HeLa), and the proliferation of the cell was observed. RESULTS: NS was highly expressed in 6 kinds of tumor cells. NS expression level in the NS-siRNA-HeLa cells was remarkably reduced, and the percentage of G_0/G_1 cells increased. The neoplasm forming ability in nude mice by the NS-siRNA-HeLa cells was decreased. CONCLUSION: NS is highly expressed among tumor cells. NS-specific siRNA inhibits the entry of the cell cycle into the S phase, and remarkably reduces the proliferation ability of HeLa cells in vitro and in vivo.

Bone Transplantation ; Dental Research ; Humans ; Tooth

Objective To review the safety and efficacy of upper-pole access percutaneous nephrolithotomy (PCNL). Methods From May 2008 to May 2010,237 cases ( 135 males and 102 females with mean age of 42 yrs) of renal or proximal ureteral calculi treated with upper-pole access PCNL were reviewed.The indications included calculus larger than 1.5 cm or impacted proximal ureteral calculi in 94 cases,calculus≥2 cm or impacted renal pelvic calculi in 26 cases,staghorn or multiple calculi in 68 cases,complex lower calyx calculi in 13 cases,upper calyx calculi not amenable to ESWL or URS in 12 cases,calculi within upper calyx diverticulum in 3 cases,combined UPJ obstruction or upper ureteral stenosis in 8 cases,morbidly obese patients in 3 cases,calculi within horseshoe kidneys in 6 cases,calculi within transplanted kidneys or ureters in 4 cases.Of the 237 cases,175 tracts (73.8％) were above the 12th rib,46were above the 11th rib,12 were below the 12th rib,4 were in the lower abdomen for renal transplant patients. Results The overall stone clearance rate with upper-pole access PCNL monotherapy was 74.3％.Additional punctures were required in 55 cases and combined ESWL in 6 cases.Total stone clearance rate at 3 months after operation was 88.2％.16 patients (6.8％) had a pleural injury.Thoracentesis was required in 8 patients,closed thoracic drainage in 5 patients,conservative treatment in 3 patients.After nephrostomy tube removed,pleural irritation symptom appeared in 12 cases (5.1％) who required symptomatic treatment.No patient had injury to the lung or other viscera.Significant bleeding requiring blood transfusion was olserved in 5 patients,while selective renal arterial embolization was required in 2 cases. Conclusions Upper-pole access offers optimal visibility,convenience for the movement of rigid nephroscope and high stone-free rate.It should be attempted in selected cages of upper ureteral calculi and complex renal calculi.

Objective To evaluate the efficacy and safety of bladder hydrodistention for the treatment of ketamine-associated bladder dysfunction.Methods Six patients were required to withdraw the ketamine and treated with bladder hydrodistention therapy and sodium hyaluronate irrigation,and medicine to pretect liver and kidney was also used.Results The biopsies of 6 cases demonstrated the cystitis through biopsy.Lower urinary tract symptoms such as urgency,thamuria and odynuria were significantly relieved after bladder installation within 30 days.The O'Leary-Sant ICSI scores and the ICPI scores reduced to 3.5 ± 1.6,2.8 ± 1.5 respectively.The functional bladder capacities increased to an anverage of (180 ± 28)ml,.2-3 times of nocturia,Qmax (14.4 ± 4.3) ml/s.All cases were followed up for 4 to 18 months.Symptoms disappeared or were significantly relieved in all patients.Conclusion Contracture of bladder might be the main presentation of ketamine-associated bladder dysfunction.Intravesical hydrodistention therapy and sodium hyaluronate irrigation could be the safe and effective therapy in the treatment of katamine-associated dysfunction.

BACKROUND:Previous studies have confirmed that radiofrequency ablation(RFA)has not only efficiently killed tumor cells,but also improved immune suppression of organism.Antigen presenting cells play an important role during anti-tumor immune reaction.Dendritic cells are most powerful antigen presenting cells.OBJECTIVE:To study the influence of RFA on OX-62,OX-6 and CD86 expression of rat peripheral blood mononuclear cells.DESIGN,TIME AND SETTING:The randomized controlled animal observation was performed at the School of Oncology of Peking University from June 2005 to March 2006.MATERIALS:A total of 12 Sprague Dawley rats were equally and randomly assigned into control and 1 -week radiofrequency groups.METHODS:2 mL peripheral blood was extracted from rats of the control group following rats were sacrificed.The left lobe of rat liver was exposed,inserted with acicular electrodes at tilted position.The acicular electrodes were spread out.The action time was 4 minutes.After RFA,the rats were given anti-infective therapy.2 mL peripheral blood was taken out after they were put to death 1 week after RFA.MAIN OUTCOME MEASURES:The following parameters were measured:pathomorphological observation on the liver of normal rats after RFA;OX-62,OX-6 and CD86 expression of peripheral blood mononuclear cells before and after RFA.RESULTS:Pathologyical examination after RFA showed the characteristic that coagulation necrosis and cellular degeneration and granulation tissue forming appeared from target center to peripheral of the target.Positive expression rate of OX-62 in rat peripheral blood in the 1-week radiofrequency group[(0.70±0.16)%]was significantly higher than in the control group [(0.34±0.08)%,P ＜0.05].No significant difference in positive expression rate of OX-6 and CD86 in the peripheral blood mononuclear cells between both groups(P＞0.05).CONCLUSION:RFA can promote the increased number of precursor Dendritic cells in rat peripheral blood,which may be contributed to improve the ability to angtigen presentation during immune response.Dai Wei-de,Doctor,Associate chief physician,Department of Ultrasound,Beijing Hospital of Ministry of Public Health,Beijing 100730,China dai.weide@126.com

Objective To conduct cloning, sequencing and expression of Trichinella spiralis ES antigen p49 gene. Methods RT-PCR was used to amplify the specific gene fragment from the total RNA of Trichinella spirais larvae. The PCR product was ligated to the T-vector and the recombinant plasmid was verified by sequencing. T-p49 and pGE-4T-3 were treated by both BamHI and XhoI. The ligation reaction was catalyzed by T4 DNA ligase. Results The p49 gene was cloned by using RT-PCR. Sequence analysis showed that the p49 gene obtained was consistent to the p49 sequence reported in the database. The expressed protein was shown as a new band at SDS-PAGE. BLAST analysis demonstrated that this p49 gene was 99% identical to the p49 gene reported and to the 43 kDa secreted glycoprotein gene in the database. Conclusion p49 gene from Trichinella spiralis larvae was cloned, sequenced and expressed.

Objective To compare and evaluate Organelle-cell fluorescence intensity ratio analysis established in this study with other techniques to determine the subcellular localization of photosensitizers. Methods CCD fluorescence microscopy imaging system was applied and a kind of special organelle probe BODIPY was selected to label Golgi body. Directly observing, pseudo-color fusing, wave-shape comparing, correlation coefficient calcula-ting and Organelle-cell fluorescence intensity ratio analysis were adopted, respectively, to study the intracellular distribution of domestic photosensitizer Hematoporphyrin monomethyl ether (HMME). Results Fluorescence distributing modes of HMME and BODIPY were similar with each other. There ware yellow space-overlap areas in the fusion image. Wave body of changing curve of gray scale value of all pixels in the straight line in two images corresponding to space coordinate was similar with each other. Correlation coefficient in each pixel between fluorescence intensity of HMME and that of BODIPY was 0.602 4. With increasing of parameter m, namely degree of organelles congregated reducing, the average fluorescence intensity ratio (J_1/J_2) of Golgi complex was decreasing, the two parameters po-ssessed obvious relativity. P

Purpose To assess the feasibility of nuclear score combined with ultrasound features on indeterminate thyroid nodules in fine needle aspiration cytological diagnosis.Methods A consecutive cohort of 119 thyroid fine-needle aspiration(FNA)specimens with indeterminate diagnosis(TBSRTC cate-gory Ⅲ-Ⅴ)and available histopathologic follow-up was evalua-ted.Original sonographic images and cytological slides of each nodule were reviewed by two experienced sonographers and two experienced cytopathologists respectively,and evaluated jointly to obtain consensus.The identification of the optimal cut-off points of simplified nuclear score and ultrasound features score for the diagnosis of malignancy or low-risk neoplasm were evalu-ated using the receiver operating characteristic(ROC)curves and the assessment of the area under the ROC curve(AUC).The specificity,sensitivity,positive predictive value(PPV)and negative predictive value(NPV)of nuclear score and US score were evaluated from crosstabs based on Cut-off points and signif-icance were calculated.The diagnostic accuracy of simplified nuclear score combined with ultrasound features was estimated by Logistic regression and ROC curve analysis.Results Nucle-ar grooves,intra-nuclear inclusions and chromatin clearing were more common in malignancy/low-risk neoplasms(P = 0.001,0.013 and 0.001 respectively).A Cut-off point of≥2 for the simplified nuclear score was sensitive for defining malignancy/low-risk neoplasm,and the sensitivity was 99.1%.Nodule height greater than width,absent halo sign,irregular margin and absent intranodular vascularization were more common in malig-nancy/low-risk neoplasms(P = 0.003,0.007,0.003 and 0.004 respectively).When the Cut-off point was set as≥5,the PPV and specificity for defining malignancy/low-risk neoplasm were 95.9%and 78.6%respectively.ROC curve analysis showed that the diagnostic accuracy of nuclear score combined with US score(0.873,95%CI = 0.777-0.970)was higher than that of nuclear score alone(0.817,95%CI = 0.687-0.948,P =0.092)or US score alone(0.758,95%CI 0.637-0.879,P =0.047).But no significant difference was detec-ted between the combined score and the nuclear score.Conclu-sion The addition of simplified nuclear score and US score to FNA cytology can increase the diagnostic accuracy of cytology in indeterminate categories.

Objective: To investigate the diagnostic value of ultrasonography.in elderly patients with gastric tumors. Methods: A total of 42 elderly patients with gastric tumors confirmed by surgery pathology underwent oral contrast-enhanced ultrasonography and electronic gastroscopy.The diagnostic effect of oral contrast-enhanced ultrasonography and electronic gastroscopy were compared based on the gold standard of surgical pathology. Results: Of the 42 elderly patients with gastric tumors, 40 patients were diagnosed by oral contrast-enhanced ultrasonography and the surgical pathology(40/42, 95.2%), with Kappa value of 0.812.And 38 cases were diagnosed by electronic gastroscopy and the surgical pathology(38/42, 90.5%), with Kappa value of 0.718(P<0.05). Based on gold standard of surgical pathology results, the accuracies of T1, T2, T3 and T4 stage by oral contrast-enhanced ultrasonography were 95.0%(38/40), 95.0%(38/40), 100.0%(40/40)and 100.0%(40/40), respectively.The sensitivity and specificity of T3 and T4 stage for oral contrast-enhanced ultrasonography reached to 100.0%.The diagnostic accuracies of N0, N1-N3, M0 and M1 by oral contrast-enhanced ultrasonography were 95.0%(38/40), 95.0%(38/40), 100.0%(40/40)and 100.0%(40/40). Conclusions Oral contrast-enhanced ultrasonography is a simple, painless and non-invasive method for the diagnosis of gastric tumors in the elderly.And its qualitative coincidence rate of the tumors diagnosis is similar to that of gastroscopy.So it can be used as the first choice for the examination of gastric tumors in the elderly.Oral contrast-enhanced ultrasonography has a high diagnostic accuracy on TNM staging of gastric tumor.When combined with electronic gastroscopy, it may provide a more reliable basis for the choice of evaluation of treatment and prognosis in elderly patients with gastric tumor.