Human platelets have distinct characters when preserved by different methods. A efficient flow cytometric assay for different preserved platelets expression of CD62p and phosphatidylserine(PS) is in dire need. Efficient flow cytometric assay for CD62p and PS expressed by preserved platelets was established and the major conditions were optimized. The platelets need not to be washed to wipe off plasma and can be labelled diredtly during the sample preparation. It is efficient for flow cytometric analysis when fresh platelet riched plasma (FPRP) was set as negative control, thrombin actived FPRP, and liquid nitrogen treated FPRP were set as positive control respectively. Gly-Pro-Arg-Pro acetate salt (GPRP) was applied to prevent platelets aggregation and fibrin formation, stabilize platelets and minimize the artificial platelets activation. This is also the key to conquer difficulty of flow cytometric quantitive analysis when platelet, Ca(2+) and plasma coexist. This flow cytometric method is specially suitable for the multi-parameter assay including PS expression for cryopreserved platelets. Minimal sample manipulation, no fixation, and GPRP application resulted in minor artifacts and good sample stability. Results suggested, this flow cytometric assay for preserved platelets is simple and efficient. In addition, the author prepared four different methods treated platelets that can be easily distinguished through this flow cytometric assay. It not only makes sure the practicability of this flow cytometric assay, but also suggests the value of the treated platelets applied in preserved platelets flow cytometric ass
Blood Platelets ; metabolism ; Flow Cytometry ; methods ; Humans ; P-Selectin ; biosynthesis ; Phosphatidylserines ; analysis ; Reproducibility of Results ; Tissue Preservation

In order to explore the factors that affect CD62p expression in platelet during the whole course of cryopreservated platelets preparation, CD62p expression of platelet was evaluated by flow cytometry assay. The whole course of cryopreservated platelets preparation in order included whole blood collection, centrifugation for fresh platelet-rich plasma preparation, addition of dimethyl sulfoxide, and freeze in -80 degrees C refrigerator and thaw in 38 degrees C water bath. Result showed that the CD62p expressed slowly from whole blood collection to addition of dimethyl sulfoxide, but expressed abruptly after freeze and thaw and it occupied 82 per cent of the whole expression. It was concluded that the whole blood collection, centrifugation for fresh platelet enriched plasma preparation and addition of dimethyl sulfoxide were optimized in the whole course, but the damage to platelets in the whole course of cryopreservation could not be avoided. It suggests that improved techniques are needed to reduce the damage to cryopreservated platelets.
Blood Platelets ; metabolism ; Blood Preservation ; methods ; standards ; Cryopreservation ; methods ; standards ; Flow Cytometry ; Humans ; P-Selectin ; biosynthesis

Objective To study the workplace violence situation of hospital setting and explore its influence on the psychological health status of nurses.Methods A descriptive and correlative design was used.By cluster sampling,932 nurses from three hospitals in Shenzhen were recruited.The Workplace Violence in Hospital Setting Questionnaire and the Symptom Checklist 90 were used to measure the status of workplace violence in hospital setting and the level of psychological health status of nurses.The data were analysed with software SPSS version 12.0.Results The overall incidence rate of workplace violence was 64.3％ with psychological violence 86.0％,physical violence 20.9％,sexual violence 11.5％.The scores of psychological health and 9 dimensions score of nurses suffering from workplace violence were significantly higher than that of nurses without suffering from workplace violence ( Z =- 9.734,P ＜ 0.01 ).The more frequence of working violence they suffered,the higher the score of psychological health of nurses they have( P ＜0.01 ),and there was significantly difference in the frequence of psychological violence,the score of psychological health and its dimension(P ＜0.01 ).The frequence of physical violence and sexual violence was no correlation with the score of psychological health ( P ＞ 0.05 ).The more frequence of working violence is,the higher scores on psychological health status of nursing staff were.( P ＜ 0.05 ).Conclusions The overall incidence rate of workplace violence was high.The psychological health status of nurses suffering from workplace violence was lower than that of nurses without suffering from workplace violence.The more frequence of working violence,the poorer the psychological health status of nurses.

OBJECTIVESTo investigate the diagnosis, therapy and prognosis of renal oncocytoma.

METHODSThe data of 26 patients of renal oncocytoma in the department of urology of the Peking University First Hospital from 1999 to 2010 were collected. Retrospective analysis of renal oncocytoma clinical characteristics of patients including pathological characteristics and long term follow-up, treatment methods, and prognosis was undergone.

RESULTSThe mean age of these patients was (55 ± 14) years, mean tumor size was (4.1 ± 1.3) cm, corresponding to a mean tumor volume of (24.0 ± 21.1) cm(3). All of these cases were unilocular unilateral. All of these 26 patients received surgical treatment, 5 cases accepted partial nephrectomy while the other cases accepted radical nephrectomy. After a median follow-up time was (47 ± 34) months (range 4 - 144 months) for 25 patients who had been followed up, there was no disease recurrence, progression or death attributed to oncocytoma.

CONCLUSIONSRenal oncocytoma is an uncommon tumor. It exhibits numerous features which are characteristic but not necessarily unique.Preoperative renal mass biopsy is the only way for pre-nephrectomy histologic diagnosis of oncocytoma.

Adenoma, Oxyphilic ; diagnosis ; surgery ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; diagnosis ; surgery ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Treatment Outcome

CD62p expression was an important monitoring parameter for preserved platelets quality. To setup an optimized flow cytometry assay for preserved platelets based on the CD62p expression on platelets, the platelet samples were collected, 0.1 mmol/L persantine and 1.1 mmol/L EDTA were added into the modified TB S used to replace PBS dilution; the methodological evaluation were carried out. Results showed that 0.1 mmol/L persantine and 1.1 mmol/L EDTA achieved to prevent platelets activation during the test procedure. The favorable negative or positive samples were prepared for check of fluorescence antibody's quality to ensure the validity of results. CD61 was used to identify platelets for assay and improve veracity of assay. The special injector was also replaced by special big syringe needle for blood collection to reduce in vitro artifacts, and the prepared sample can be steady-going for 48 hours at 4 degrees C after fixed by 1% paraformaldehyde. It is concluded that this flow cytometry assay for CD62p positive platelets is simple and efficient.
Blood Platelets ; chemistry ; Blood Preservation ; Flow Cytometry ; methods ; Humans ; P-Selectin ; blood

OBJECTIVETo study the effects of drug plasma of musk and olibanum (DP-M&O) on the release of inflammatory cytokines from monocytes and the expressions of the proteins associated with inflammation of prostatic or endothelial cells induced by prostate antigen (PAg) stimulation.

METHODSWe prepared DP-M&O using SD rats and monocytes and PAgs using BALB/c mice. We pre-treated the monocytes with DP-M&O at the gradient concentrations of 0, 2.5, 5, 10, and 20% for 1 hour, activated them with PAgs, and then cultured them for 96 hours, followed by detection of the release of inflammatory cytokines. We co-cultured the prostate RWPE-1 cells with the endothelial EA. hy926 cells, pre-treated them with the same gradient concentrations of DP-M&O as above for 1 hour, activated with PAgs, and cultured for 96 hours. Then we determined the expression levels of the proteins associated with inflammation of RWPE-1 and EA. hy926 cells by Western blot.

RESULTSDP-M&O decreased the levels of TNF-alpha, IL-1beta, IL-6, and IL-8 and increased that of IL-10 in a concentration-dependent manner. Significant differences were found between the 20% P-M&O and PAg groups in the release of the inflammatory cytokines TNF-alpha (70.8 +/- 22.3 vs. 277.1 +/- 65.5, P < 0.01) , IL-113 (277.5 +/- 22.6 vs. 630.4 +/- 89.7, P <0.01), IL-6 (232.7 +/- 62.7 vs. 994.2 vs. 182.3, P < 0.01), IL-8 (227.3 +/- 79.2 vs. 769.3 +/- 284.1, P < 0.01), and IL-10 (640.2 +/- 201.2 vs. 271.1 +/- 55.8, P < 0.01). Compared with the PAg group, the 10 and 20% P-M&O groups showed remarkable decreases in the protein expression of MCP-1/CCL2 in the RWPE-1 cells (1.12 +/- 0.34 vs. 0.56 +/- 0.11 and 0.34 +/- 0.08) and that of VCAM-1 in the EA. hy926 cells (0.94 +/- 0.22 vs. 0.52 +/- 0.17 and 0.38 +/- 0.12) (P < 0.05 or 0.01).

CONCLUSIONThe compatibility of musk and olibanum can decrease the expression of MCP-1/CCL2 in prostate cells and VCAM-1 in vascular endothelial cells, blocking the adhesion of leucocytes and suppressing inflammatory response.

Animals ; Blotting, Western ; Cytokines ; metabolism ; Endothelial Cells ; drug effects ; metabolism ; Fatty Acids, Monounsaturated ; pharmacology ; Frankincense ; pharmacology ; Inflammation ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; Male ; Mice ; Mice, Inbred BALB C ; Monocytes ; drug effects ; metabolism ; Prostate ; cytology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism

Objective:Carcinoma of bladder is the most common malignancy in the urinary system in China.Most patients with this disease had non-muscle invasive bladder cancer (NMIBC)at the time of diagnosis.Radical cystectomy was indicated for patients with high risk or refractory NMIBC.We aimed to investigate the overall survival and disease-specific survival and related influence factors in patients un-dergoing radical cystectomy for pathological non-muscle invasive bladder cancer.Methods:From Jan. 2006 to Dec.2012,a total of 164 patients with pathological non-muscle invasive bladder cancer under-went radical cystectomy in Peking University First Hospital.Clinical data were retrospectively collected. Incidence of lymph node metastasis and disease recurrence were calculated.The risk factors of disease re-currence were analyzed.Kaplan-Meier plots were used to estimate the overall survival and cancer-specific survival.Multivariate Cox regression analysis was used to evaluate the prognostic factors for survival.Re-sults:Of all the patients included,159 had T1 disease,and 5 had CIS only.The median follow-up dura-tion was 46.5 months (range:7 -99 months).Fourteen patients were lost during the follow-up.Lymph node metastasis was noted in 6 patients (3.7%),4 patients had N1 disease,one patient had N2 di-sease,and one patient had N3 disease.Disease recurrence occurred in 16 patients (9.8%).The most common recurrence sites were the liver,bones,and lungs.The 5-year overall survival and disease-spe-cific survival for all the patients were 85% and 91%,respectively.The patients who underwent pelvic lymph node dissection showed a better prognosis in terms of disease-specific survival than those without (P =0.012).Patients with recurrence harbored a significant poorer survival (P <0.001).According to univariate Cox regression analysis,whether lymph node dissection was performed was an independent risk factor of disease recurrence (P =0.050,OR =2.695,95%CI 0.999 -7.271).In COX regression mo-del,age (P =0.008,OR =1.071,95%CI 1.018 -1.126)and whether lymph node dissection was performed (P =0.011,OR =3.385,95%CI 1.329 -8.621)were related to disease-specific survival. Conclusion:Patients with pathological non-muscle invasive bladder cancer underwent early radical cys-tectomy have a favorable prognosis,and bilateral pelvic lymph node dissection is essential for this proce-dure as it gains a survival benefit for the patients.

OBJECTIVETo investigate the effects of the combination of musk and olibanum on the tight junction protein expressions in prostatic epithelial cells of normal and chronic prostatitis (CP) rats.

METHODSEighty male SD rats were randomly divided into 8 groups of equal number: normal control, normal musk, normal olibanum, normal musk + olibanum, CP model control, CP model musk, CP model olibanum, and CP model musk + olibanum. At 60 days after modeling, the rats in the control, musk, olibanum, and musk + olibanum groups were treated intragastrically with normal saline, musk (0.021 g per kg body weight per day), olibanum (1.05 g per kg body weight per day), or musk + olibanum respectively, all for 3 days. Then, all the rats were sacrificed and their prostate tissues harvested for detection of the expressions of the tight junction proteins Claudin-1, Claudin-3, Occludin, and ZO-1 in the prostatic epithelial cells by immunohistochemical staining.

RESULTSIn the CP models, only the expression of Claudin-1 was significantly increased. In the normal rats, the expression of Claudin-1 was remarkably upregulated after treated with musk (824.6 ± 393.3, P < 0.05), olibanum (982.0 ± 334.0, P < 0.05), and musk + olibanum (1088.1 ± 640.2, P < 0.01); that of Claudin-3 was elevated markedly by olibanum (1 009.5 ± 243.6, P < 0.05) and insignificantly by musk (597.5 ± 80.7), but the increasing effect of olibanum was reduced by musk + olibanum (678.4 ± 255.1). No statistically significant differences were found in the expression of Occludin among the rats treated with musk (693.0 ± 424.8), olibanum (732.1 ± 302.0), and musk + olibanum (560.2 ± 202.3), or in that of ZO-1 in the animals treated with musk (290.0 ± 166.8) and olibanum (419.7 ± 108.1), but the latter was markedly decreased in the musk + olibanum group (197.7 ± 98.2, P < 0.05). In the CP rat models, both the expressions of Claudin-1 (823.0 ± 100.1, P < 0.01) and Occludin (1160.0 ± 32.2, P < 0.05) were significantly increased. The expression of Claudin-1 was remarkably down-regulated by musk (764.9 ± 179.0), olibanum (468.4 ± 220.4), and musk + olibanum (335.1 ± 204.0) (all P < 0.05), but that of Claudin-3 up-regulated by musk (744.6 ± 94.5) and olibanum (700.1 ± 223.7) (both P < 0.05). The expression of Occludin was reduced by musk (615.0 ± 221.0), olibanum (749.6 ± 321.7), and musk + olibanum (505.8 ± 523.7), while that of ZO-1 increased by olibaum (443.2 ± 44.9) and decreased by musk + olibanum (213.5 ± 24.9, P < 0.05).

CONCLUSIONIn physiological and pathological conditions, the combination of musk and olibanum acts on the expressions of tight junction proteins in prostate epithelial cells in a selective and dual-targeting manner, promoting their permeability by down-regulating the expression of ZO-1 and maintaining their structural stability by regulating the expressions of Claudin-1, Claudin-3, and Occludin.

Animals ; Claudins ; metabolism ; Down-Regulation ; Epithelial Cells ; drug effects ; Fatty Acids, Monounsaturated ; chemistry ; Frankincense ; chemistry ; Male ; Occludin ; metabolism ; Prostate ; cytology ; Prostatitis ; Rats ; Rats, Sprague-Dawley ; Tight Junction Proteins ; metabolism ; Up-Regulation

Variant pulmonary vein anatomy (PVA) has been reported to influence the recurrence of atrial fibrillation (AF) after radiofrequency ablation.However,the effects of PVA on AF in patients undergoing cryoballoon ablation (CBA) remain unknown.The present study aimed to examine the impact of PVA on the long-term outcome of CBA for AF.A total of 78 patients (mean age 60.7±10.9 years,64.1％ males) with symptomatic and drug-refractory paroxysmal AF were enrolled in the study.Left atrium (LA) and PVA acquired at computed tomography angiography (CTA) were reconstructed with CARTO(R) 3 SYSTEM.Patients were routinely evaluated by 24-hour Holter monitoring following CBA.Cox regression was used to detect the predictors of AF recurrence after CBA.The results showed abnormal PVA in 30 patients (38.5％) and 18 patients (23.1％) had left common PV (LCPV).Electrical pulmonary vein isolation was achieved in all patients.After a mean follow-up of 689.5±103.8 days,it was found that patients with abnormal PVA had similar AF recurrence rate to those with normal PVA (26.7％ vs.25.0％,P=0.54),and there was no significant difference in AF recurrence rate between LCPV patients and non-LCPV patients (33.7％ vs.23.3％,P=0.29).Cox regression analysis showed that AF duration (72.9±9.0 vs.42.3±43.2 months,HR 1.001;95％CI 1.003-1.014;P＜0.001) and cryo-applications of right-side PVs (3.0±1.6 vs.4.7±1.7,HR 0.661;95％ CI 0.473-0.925;P=0.016) were independent predictors of freedom from AF,but PVA was not identified as a predictor of long-term success.In conclusion,the variant PVA cannot significantly influence the long-term outcome of AF patients undergoing CBA;longer AF duration and less cryo-applications of right-side PVs are associated with higher AF recurrent rate.

OBJECTIVETo investigate the expression of miR-9 in B lymphocytes, B cell lymphoma and classical Hodgkin's lymphoma (cHL) cell lines and its significance.

METHODSCD19(+) B lymphocytes were sorted from normal lymph node by magnetic beads. Total cellular micro-RNA was extracted from cHL cell line L428, B cell lymphoma cell lines Ly1 and Ly10 (diffuse large B cell lymphoma), Raji cells (Burkitt's lymphoma) and CD19(+) B lymphocytes, respectively. These micro-RNAs were separately transformed into cDNA by reverse transcription. The expression levels of miR-9 were measured by fluorescence quantitative PCR. In situ hybridization was used to detect the expression of miR-9 in cell lines.

RESULTSThe expression of miR-9 was high in L428 cells (104.44 ± 1.61), and low in cell lines of B cell lymphoma (Ly1: 2.17 ± 0.38; Ly10: 1 ± 0.015; Raji: 2.65 ± 0.89), and extremely low in CD19(+) B lymphocytes (0.0026 ± 0.00040). Compared with that in the other cell lines, the expression of miR-9 in L428 cells was statistically significant (P < 0.05). miR-9 localized in the cytoplasm diffusely and strongly in L428, but scattered and slightly with some prominent distribution around the nuclear membranes in Ly1 and Ly10, and only weakly in Raji.

CONCLUSIONSmiR-9 highly expressed in cHL cell line and might be a molecular marker for diagnosis and treatment of cHL.

B-Lymphocytes ; metabolism ; Cell Line, Tumor ; Cell Lineage ; Hodgkin Disease ; metabolism ; pathology ; Humans ; Lymphoma, B-Cell ; metabolism ; pathology ; MicroRNAs ; metabolism