[Development and prospective validation of an ischemic risk score for acute chest pain patients with normal high-sensitivity troponin I levels and without obvious ST-segment deviation].

Objective: To develop and prospectively validate a risk score for acute chest pain patients with normal high-sensitivity troponin I (hs-TnI) levels and without obvious ST-segment deviation in China. Methods: Chest pain patients admitted to the emergency department of Beijing Anzhen Hospital from September 2014 to July 2015 were enrolled. Baseline characteristics of patients met inclusion criteria including normal hs-TnI levels and without obvious ST-segment deviation were included. The endpoint (major adverse cardiovascular events) was a composite of acute myocardial infarction, percutaneous coronary intervention, coronary artery bypass graft, and all-cause death within 3 months after initial presentation. Predictors were screened and used to develop the risk score model by logistic regression analysis in a retrospective cohort. Then, the risk score model was evaluated in a prospective validation cohort. Results: The study population of derivation cohort included 1 735 consecutive chest pain patients. Finally, 1 030 eligible patients were enrolled. Multivariate regression analysis defined five independent predictors: male gender (ß=0.88); history of chest pain (ß value of moderate and high suspicion of coronary heart artery was 2.70 and 3.51 respectively); electrocardiogram (ß=0.84); ≥60 years old (ß=0.51) and ≥3 risk factors (ß=0.85).The range of weighted score was set as 0-13. The area under a receiver operating characteristic (ROC) curve was 0.75 (95%CI 0.72-0.78) in the final model. Major adverse cardiovascular events rates increased in proportion to score increase (P<0.01). The internal validity used bootstrap technique showed the same predictor factors as the final model, and its area under a ROC curve was 0.75(95%CI 0.72-0.78).MACE rates in the low risk group (score 0-3), intermediate risk group (score 4-7), and high risk group (score 8-13) were 1.3% (1/77) ,19.0% (22/116) ,and 42.2% (122/289) in the prospective validation cohort, respectively (P<0.01). Conclusion: The developed ischemic risk score is feasible for risk stratification of acute chest pain patients with normal hs-TnI and without obvious ST-segment deviation, this score might be helpful to the decision making of treatment and management strategies for these patients.

MicroRNA-99b inhibits NSCLC cell invasion and migration by directly targeting NIPBL.

OBJECTIVE: Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related death worldwide. microRNAs (miRNAs) have been confirmed as vital regulators of multiple tumors, including NSCLC. The aim of the current study was to explore the biological mechanisms of miR-99b in NSCLC progression. PATIENTS AND METHODS: NSCLC tissues and adjacent matched human non-neoplastic lung tissues used in this study were collected from 50 cases of NSCLC patients. The expression of miR-99b and NIPBL in NSCLC tissues and cell lines (A549, NCI-H460, NCI-H1299 and SPC-A1) were determined by real-time-polymerase chain reaction (qRT-PCR). The NIPBL protein level was measured by Western blot. Dual-Luciferase reporter, Western blotting and qRT-PCR were carried out to verify the potential target of miR-99b. Transwell assay was used for investigating miR-99b effect on cell migration and invasion in NSCLC cells. RESULTS: The results of qRT-PCR indicated that the expression of miR-99b was downregulated in the NSCLC tissues and cell lines. Overexpression of miR-99b could significantly inhibit the invasion and migration capacities in NSCLC cells. Furthermore, we also determined that NIPBL was a direct target of miR-99b. Additionally, we found NIPBL was implicated in the suppressive effects on NSCLC cell invasion and migration mediated by miR-99b. CONCLUSIONS: In summary, miR-99b exerted anti-tumor functions in NSCLC via regulation of NIPBL, suggesting that miR-99b/NIPBL axis may be novel biomarkers for NSCLC treatments.

The onset of nonlinearity in cosmological structure.

We discuss the progression of growth of cosmological structure, from the quasilinear evolution of nearly Gaussian fluctuations on large scales into highly non-Gaussian, strongly nonlinear structure on small scales. A systematic development in perturbation theory describes the first departures from homogeneity but fails to reproduce the fully nonlinear results. Physical insight, conceptual models, and symmetries are useful in the strong clustering regime. A phenomenological model with input information from the quasilinear regime provides enticing results for the strongly nonlinear regime.

Effects of high-energy shock waves on murine renal cell carcinoma.

The effects of high-energy shock waves (HESW) on a murine renal cell carcinoma (RenCa) was investigated. In vitro exposure of tumor cells to HESW resulted in a dose-dependent reduction in cell viability as determined by trypan blue dye exclusion, plating efficiency, growth curve, and soft agar clonogenic assays. Activity of lactic dehydrogenase (LDH) was detected in the supernatant after the HESW treatment due to cellular destruction, and a dose-dependent increase in cytocidal effect was demonstrated. Ultrastructural changes with swelling and distorted cristae of mitochondria, vacuolation, ribosomal lysis, and chromatinolysis were observed in HESW-treated RenCa cells. Flow cytometric (FCM) study revealed that DNA content of RenCa cells diminished after 200 HESW treatment, and RNA content of tumor cells decreased markedly after 400 HESW treatments. Partial or complete inhibition of tumor growth was shown in both animal modalities of subcutaneous inoculation and intravenous injection with sequential lung metastases. This study stressed again that HESW may play a role in combinational protocol for the treatment of human renal cell carcinoma in certain circumstances.

Effect of capsaicin on micturition and associated reflexes in chronic spinal rats.

The role of capsaicin-sensitive bladder afferents in micturition was studied in unanesthetized chronic spinal rats. Reflex voiding in response to tactile stimulation of the perigenital region appeared 5-9 days after spinal cord injury (SCI) whereas voiding induced by bladder distension occurred 2-3 weeks after SCI. The frequency and amplitude of reflex bladder contractions recorded under isovolumetric conditions were similar in chronic spinal and urethane-anesthetized CNS-intact rats. However, cystometrograms (CMGs) performed 6-8 weeks after SCI revealed that the chronic spinal rats had larger bladder capacities (1.86 ml) than CNS-intact rats (0.48 ml) and also exhibited multiple, small-amplitude, nonvoiding bladder contractions that were not detected in CNS-intact rats. Administration of capsaicin (50 mg/kg s.c.) acutely (onset 14-40 min) suppressed reflex bladder activity induced by bladder distension or by perigenital stimulation in chronic spinal animals. However, pretreatment of chronic spinal rats with capsaicin (125 mg/kg s.c.) 4 days before the experiment did not depress voiding reflexes or change bladder capacity but did eliminate the nonvoiding contractions. Inhibition of reflex bladder contractions by mechanical stimulation of rectoanal canal or the uterine cervix-vagina was not altered by pretreatment with capsaicin. These data indicate that capsaicin-sensitive bladder afferents are not essential for the initiation of reflex micturition in chronic spinal rats. However, these afferents do contribute to hyperactivity of the bladder during the filling phase of the CMG. Thus, capsaicin-sensitive bladder afferents should be evaluated as possible targets for the pharmacological treatment of bladder hyperreflexia in patients with SCI.

A genetic study of human interferon-alpha-induced repair of DNA damage in hepatitis B patients.

In vitro treatment with human interferon-alpha (HuIFN-alpha) of hepatitis B virus-infected peripheral lymphocytes from 17 hepatitis B patients induced a decrease in the frequency of sister-chromatid exchanges (SCE). There was a significant difference in mean SCE frequencies between the HuIFN-alpha-treated patients and the control group, but not between acute and chronic hepatitis B patients treated with HuIFN-alpha.

Prevalence of cervical cancer and feasibility of screening in rural China: a pilot study for the Shanxi Province Cervical Cancer Screening Study.

For cervical cancer screening to be feasible in developing countries, it must be accurate, inexpensive, and easy to administer. We conducted a pilot study in rural Shanxi Province, People's Republic of China, to determine disease prevalence and study feasibility in preparation for a large-scale comparative trial of 6 screening tests. One-hundred and thirty-six nonpregnant women with no history of hysterectomy, pelvic radiation, or Papanicolaou tests were screened in a rural clinic. Ten percent of the women enrolled reported abnormal vaginal bleeding and 45% reported abnormal vaginal discharge. The tests were the Papanicolaou test (both conventional and ThinPrep), a self-administered swab test by Hybrid Capture II for high-risk human papillomavirus (HPV), a test for high-risk HPV from residual PreservCyt medium, fluorescence spectroscopy, and visual inspection of the cervix by a clinician. All women also underwent colposcopy and biopsies as the reference standard. Biopsies showed 12 of 136 women had >/= high-grade squamous intraepithelial lesions (HGSIL). Screening was completed in 5 half-day sessions, the procedures went smoothly, and local cooperation was enthusiastic. Disease prevalence in Xiangyuan and Yangcheng Counties, Shanxi Province, can be estimated at 8.8% (95% CI, 4.5% to 15.0%). Screening 1000-2000 patients would be sufficient to detect a 10% difference in accuracy between diagnostic tests. The proposed large-scale trial is feasible.

[Testicular morphological and biochemical changes after Fowler-Stephens orchiopexy in rats].

Fowler-Stephens orchiopexy is the most common method of treating high-positioned undescended testes. Its success rate has been reported to be as high as 50-70% based on palpation or in a few circumstances on biopsy. Although it is convenient to evaluate testicular function by palpation and/or biopsy, this method is very subjective and is not scientific enough. To determine testicular function more precisely and objectively, we performed the Fowler-Stephens orchiopexy on Sprague-Dawley rats and observed the morphological and biochemical changes, including assays of LDH, SDH and the testosterone level. In our morphological study, with the use of Fowler-Stephens orchiopexy, only 17% (3/18) of the rat tests could be salvaged. The others revealed either necrosis or fibrosis. Testicular LDH, checked at the 4th postoperative week, revealed 0.62 +/- 0.04 U/mg which was statistically different (p less than 0.05) from that of the control group and the hemicastrated group (0.77 +/- 0.05 U/mg and 0.76 +/- 0.07 U/mg, respectively). The SDH obtained from the testes also revealed significant differences between the study group and the control and hemicastrated groups. Values obtained were 2.67 +/- 0.15 mU/mg, 3.77 +/- 0.4 mU/mg and 3.77 +/- 0.33 mU/mg, respectively (p less than 0.05). Using electrophoresis, 3 out of 18 rats had typical X bands, which is the classical picture of a normal mature testis. In contrast, the others showed faint X bands at the 2nd postoperative week, which subsequently faded thereafter. During testicular ischemia, the Leydig cells are more resistant than the Sertoli cells and the germinal cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Cytomorphometric and flow cytometric analysis of bladder transitional cell carcinoma using acridine orange fluorochrome: correlation with histopathology and clinical behavior.

Cell material from 79 cases of archival paraffin-embedded tumor specimens of newly diagnosed transitional cell carcinomas of the urinary bladder was studied retrospectively using rapid flow cytofluorometric DNA analysis. The male to female ratio was 62/17. The mean age at diagnosis was 65.3 +/- 10.8 years. Clinical characteristics, including survival, recurrence and progression were closely related to the histopathologic grading and tumor staging. The tumors were classified as diploid (DI = 0.9-1.1) or aneuploid. Total aneuploid occurrence was 46%. The aneuploid frequency for the various tumor grades was 25% for grade 1, 37% for grade 2 and 78% for grade 3. Stages Ta (O) and Tl (A) tumors differed from muscle-invasive tumors (T2-4) in that they had a lower frequency of aneuploid occurrence (34%, 41% vs 50%, 73%, and 75%). Multiple aneuploid stem cell lines had no influence on tumor grade or stage. The DNA ploidy was closely related to the five-year survival rate, the probability of tumor recurrence and progression. On the contrary, the modal nuclear size of the tumor cells had no correlation with histopathology or the clinical characteristics. Bladder tumors of an intermediate grade (grade 2) may be subdivided into two groups with different outcomes on the basis of flow cytometric characteristics. It is concluded that DNA flow cytometry can serve as an additional prognostic parameter for bladder cancer patients in addition to conventional histopathology.

Verapamil modulation of multidrug resistance in renal cell carcinoma.

BACKGROUND AND PURPOSE: Renal cell carcinoma (RCC) is well known for its chemoresistance. The membranous p-glycoprotein (gp-170) is believed to be highly correlated with multidrug resistance (MDR) of cancer cells with energy-dependent pumping efflux of anticancer drugs. Verapamil, a calcium antagonist, inhibits the efflux function of gp-170 and cytoskeletal transportation. The aim of this study was to determine the effect of verapamil on gp-170 expression and intracellular drug accumulation in RCC tumor cells and the modulation of cytotoxicity of various chemotherapeutic drugs on native RCC cell lines and acquired MDR sublines by verapamil. METHODS: Using cultured cell lines of RCC and their MDR sublines as target cells, the effect of verapamil on gp-170 expression was analyzed by immunofluorescence flow cytometry. The influence of verapamil on intracellular drug accumulation in RCC tumor cells was measured by autofluorescence flow cytometry. The modulation of verapamil on cytotoxicity of various chemotherapeutic drugs on native RCC cell lines and acquired MDR sublines was analyzed by the methyl tetrazolium method. RESULTS: From flow cytometric measurement, the expression of gp-170 was significantly decreased in A704 and Caki-1 tumor cells after verapamil treatment. The uptake of adriamycin and maintenance of intracellular drugs were also significantly increased following verapamil treatment in RCC8701 tumor cells. These effects were sustained for as long as 8 hours after verapamil withdrawal. The cytotoxicity of adriamycin and epirubicin on RCC8701 and its MDR subline tumor cells was markedly intensified by verapamil. The verapamil modulation of cytotoxicity was in an immediate-reaction pattern and was dose-dependent, with synergistic effects. Long-term treatment was more effective than short-term treatment in RCC MDR sublines. The cytotoxicity of vinca alkaloid (vinblastine) and alkylators (carboplatin) was also enhanced by verapamil. CONCLUSIONS: These results suggest that verapamil plays an important role in the circumvention of native and acquired chemoresistance of RRC because it suppresses membranous gp-170 expression and cytoplasmic drug transportation.

Pathogenesis of ischemia reperfusion injury of the kidney after transient renal arterial clamping in rats.

Renal function can be severely impaired through injuries sustained after both short and prolonged periods of complete ischemia. The magnitude of renal dysfunction resulting from these conditions and their reversibility depend on the duration of anoxia. In this study, we used a Sprague-Dawley rat model (5 to 7 rats in each group) to study the pathogenesis of short-term ischemia (30, 60, and 120 min)/reperfusion (2, 4, 24 h, 1 wk, and 3 wk) injury of the kidney under warm (room temperature) or cold (4 degrees C) conditions. Ischemia was induced by clamping the renal artery. Changes in kidney weight, histopathology, concentrations of serum thromboxane and leukotriene, and tissue malonyldialdehyde (MDA) concentration, numbers of apoptotic bodies, and p53 expression in the kidney were compared with those of sham-operated rats. The results showed that the immediate increase in kidney weight due to inflammatory swelling was associated with simultaneous elevation of serum thromboxane and leukotriene levels. The changes in mediator levels were closely related to the duration of ischemia and temperature. Histologic structures were preserved better when renal artery clamping was done at 4 degrees C. MDA peroxidation products from the ischemic tissue prominently increased 1 week following ischemia; this paralleled a secondary increase in leukotriene levels. Flow cytometric detection of p53 oncoprotein showed a marked increase at 1 week following ischemia, which was accompanied by the development of apoptotic bodies in ischemic tissues. These changes were also closely related to the ischemic time and temperature during ischemia. This animal model may be useful for future studies of the prevention of ischemia/reperfusion injury of the kidney and for selection of effective antioxidants.

Integrated bioinformatics analyses identify dysregulated miRNAs in lung cancer.

BACKGROUND: Lung cancer is the most common cause of cancer-related death worldwide. MicroRNAs (miRNAs) play important roles in various biological processes, including cell development, proliferation, differentiation and apoptosis. MATERIALS AND METHODS: In the current study, using miRNA expression profiles from the Gene Expression Omnibus (GEO) database, we used three independent tests: Wilcox test, t-test and Fisher's exact test to investigate miRNA's involvement in lung carcinogenesis. RESULTS: Ten differentially expressed miRNAs were identified. Among them, miR-675 drew specific attention. Ingenuity pathway analysis of its target genes revealed its impact on cell death and cell cycle. It is possible that miR-675 contributes to the pathogenesis of lung cancer through its down regulation of the tumor suppressor gene RB1. CONCLUSIONS: Our results suggest miR-675 may serve as a potential therapeutic target of lung cancer.

Transitional cell carcinoma of the urinary bladder in patients under 40 years of age.

Transitional cell carcinoma of the bladder is the most common neoplasm of the urinary tract but it occurs infrequently in younger patients. An analysis of 25 patients aged less than 40 years showed that the clinical course and prognosis were relatively benign. It is suggested that management and follow-up should be the same as in older patients with urothelial carcinoma.

Transitional cell carcinoma of the kidney with extension into the inferior vena cava.

A case of transitional cell carcinoma of the kidney, invading the inferior vena cava, is presented. Preoperative diagnosis is almost impossible and frozen section is necessary to decide the type of surgery. Radical surgery with removal of thrombi was performed in our patient but was obviously not helpful. The value of aggressive surgery is not clear. This may be the 6th case in the literature.

Adult Wilms' tumor.

Two well-documented cases of Wilms' tumor in adult patients are reported. Surgery remains the treatment of choice in stage I cases with favorable cell differentiation. Adjuvant therapy with chemotherapy or radiation may be necessary in patients beyond stage I or when the cell differentiation is more anaplastic. Actinomycin D is essential and should be included in the chemotherapeutic regimen whenever it is possible.

Spontaneous retroperitoneal hemorrhage from kidney causes.

Spontaneous retroperitoneal hemorrhage is an uncommon entity and even rarer when the underlying cause is from the kidney. Renal tumors comprise the majority of atraumatic kidney rupture. Renal cell carcinoma and angiomyolipoma are the most common diseases in this group. Oral anticoagulant therapy and hemodialysis could be responsible for a few cases. In 3 reported cases no pathological explanation could be found. With the help of modern facilities, diagnosis can be made preoperatively and conservative surgery is indicated in these patients. However, nephrectomy is the treatment of choice for patients presenting with shock as the initial symptom or solid renal mass with perirenal hematoma.

The effect of methylene blue-sensitized photodynamic treatment on bladder cancer cells: a further study on flow cytometric basis.

The cytotoxic effect mediated by methylene blue-sensitized photoinactivation on bladder cancer cells has been well known for years, but the actual mechanism is still not clear. Specific fluorescence stains were used in a multiparameter approach. The changes in cellular size, granularity, protein content, RNA and DNA were analyzed by flow cytometry. The results showed that, in addition to cell size, all the other parameters, including cellular granularity, protein, RNA and DNA, diminished following photodynamic therapy. The cytotoxic efficacy is closely related to time of stain and illumination. The DNA and cell cycle were affected simultaneously with decreased DNA content and arrest in the G0/G1 phase. These multistage processes occurred synchronously after the phototherapy. Our results suggest that the methylene blue-sensitized photooxidation of bladder cancer cells is carried out by multimedia reactions through which the intracellular proteins (enzymes in mitochondria), RNA (ribosomes) and DNA (chromatin) decrease with loss of cytoplasmic granularity.

Photoinactivation of bladder tumor cells by methylene blue: study of a variety of tumor and normal cells.

Human bladder cancer cell lines, J82, Yen-87, Shen-87 and Zoa-88, and murine bladder cancer cell lines, MBT-2 and M1660, were used as target cells for dye-sensitized photoinactivation study in using methylene blue. Normal fibroblast cells, FB-1 and FB8490, were used as control group. The cytoplasmic activity of lactic dehydrogenase, soft agar clonogenic assay, and in vivo tumor growth, survival rate and tumor taking rate with or without photoinactivation were monitored and compared between different cell lines. Efficacy of photoinactivation was time-related and more than 90 per cent of cytotoxicity could be obtained within 60 minutes of illumination. The plateau of cytotoxicity curve could be achieved after staining for 30 minutes by methylene blue under the same illumination time. Normal fibroblasts had the same features with cancer cells. Photoinactivation of tumor cells showed significant inhibition of tumor growth and tumor taking rate in experimental animals. Survival rate was also significantly prolonged in the animals with tumor cells receiving photoinactivation. These results suggest that methylene blue-sensitized photoinactivation may be useful as an adjuvant photochemotherapy for superficial bladder cancer.

DNA ploidy, S-phase fraction and cytomorphometry in relation to survival of human penile cancer.

The prognostic significance of DNA ploidy, DNA index, S-phase fraction (SPF) and median nuclear size was studied in 11 patients with squamous cell carcinoma of the penis. These patients have been followed for a minimum of 7 months after diagnosis. Nuclear DNA content was determined by flow cytometry from paraffin-embedded tissue. Patients with DNA diploid cancer (n = 7, 64%) had a better survival rate than patients with aneuploid cancer, and a small SPF was associated with a favorable outcome. A statistically significant relation between median nuclear size and survival could be demonstrated. Small modal nuclear size associated with poorer prognosis. There was a significant difference in survival between metastatic and nonmetastatic groups of tumors during the follow-up period. This study suggests that flow cytometric determination of nuclear DNA ploidy from paraffin-embedded samples in penile cancer does add an additional prognostic determinant in addition to the clinical staging of tumors.

Identification, purification, and characterization of a protein activator (PA28) of the 20 S proteasome (macropain).

A protein that greatly stimulates the multiple peptidase activities of the 20 S proteasome (also known as macropain, the multicatalytic protease complex, and 20 S protease) has been purified from bovine red blood cells and from bovine heart. The activator protein was a single polypeptide with an apparent molecular weight of 28,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and had a native molecular weight of approximately 180,000. This protein, which we have termed PA28, regulated all three of the putatively distinct peptidase activities displayed by each of two functionally different forms of the proteasome. This regulation usually included both an increase in the maximal reaction velocity and a decrease in the concentration of substrate required for half-maximal velocity and indicated that PA28 acted as a positive allosteric effector of the proteasome. PA28 failed, however, to stimulate the hydrolysis of large protein substrates such as casein and lysozyme. These results suggested that the hydrolysis of protein substrates occurred at a site or sites distinct from those that hydrolyzed small peptides and that the regulation of the two processes could be uncoupled. Evidence for direct binding of PA28 to the proteasome was obtained by glycerol density gradient centrifugation. PA28 may play an important regulatory role in intracellular proteolytic pathways mediated by the proteasome.