Volume 93  ·   Issue 42  ·  2013

To evaluate the efficiency of O′Leary-Sant problem index and symptom index (OPSI) in the diagnosis of interstitial cystitis (IC).

A total of 82 patients, initially diagnosed as IC, with urinary frequency, urgency and pain on pubic area during bladder filling phase from January 2008 to March 2013 were enrolled.OPSI was used to evaluate their symptoms.They were eventually divided into IC and non-IC groups according to the clinical diagnosis.The scores of OPSI, interstitial cystitis problem index (ICPI) and interstitial cystitis symptom index (ICSI) were compared between two groups.And the ROC curve was plotted to screen the diagnostic specificity and sensitivity.

Among them, 58 were diagnosed with IC and the rest with non-IC.The average score of OPSI for groups IC and non-IC were 24.7±6.9 and 23.2±7.8 respectively (P=0.403).And the ICPI score for two groups were 11.7±3.2 and 10.2±3.7 (P=0.070) and the ICSI score 13.0±4.0 and 13.1±4.3 (P=0.975) respectively.No significant differences existed between two groups.If an OPSI score of 12 or greater for diagnosing IC, the sensitivity was 98% and specificity 8%.Because of a high sensitivity, OPSI was a useful indicator for screening IC patients from others.In the ROC curve, acreage under curve for OPSI, ICPI, ICSI were 0.548±0.071, 0.622±0.069 and 0.492±0.070 respectively.These three indices were not good indicators for diagnosing IC.

The O′Leary-Sant symptom index and problem index has a high sensitivity but a low specificity in the diagnosis of IC.Therefore it should not be used to differentiate interstitial cystitis.A cutoff OPSI value of 12 may be used for differentiation.

To explore the video-urodynamic characteristics and management in complete cervical and thoracic spinal cord injury patients.

Video-urodynamic examination was performed in 113 patients with complete cervical and thoracic spinal cord injury from January 2008 to May 2010.And their characteristics, managements and 3-year follow-up outcomes were reviewed and analyzed retrospectively.

Among them, there were detrusor overactivity (n=82, 72.57%), detrusor areflexia (n=31, 27.43%), detrusor external sphincter dyssynergia (n=71, 62.83%), low-compliance (n=59, 52.21%), reflux (n=6, 5.31%) and ultrasonic uronephrosis (n=12, 10.62%).According to the result of the video-urodynamic examination, detrusor overactivity patients chose oral anticholinergic agents, botulinum toxin type A injection method, detrusor areflexia patients chose regular intermittent catheterization for bladder training.During a 3-year follow-up, there were cystic calculus (n=2) and urinary infection (n=7).No special complication occurred.

The video-urodynamic characteristics vary in complete cervical and thoracic spinal cord injury patients.The major symptoms include detrusor overactivity, detrusor external sphincter dyssynergia and low-compliance bladder.Proper management is essential for protecting upper urinary tract and preventing urinary system complication.

To evaluate the effects of chronic hyperlipidemia on bladder function and its possible mechanisms and examine the functional and histological changes of bladder in a diabetic hyperlipidemic model.

A total of 30 male New Zealand white rabbits (2.5 to 3.0 kg) were numbered sequentially and randomly divided into 2 groups (hyperlipidemic group and control group).Then urodynamic test and detrusor strip contraction study were performed to assess bladder function.Histological studies of bladder and internal iliac arteries were performed with hematoxylin and eosin staining.The ultrastructure of bladder detrusor tissue was examined by transmission electron microscopy.Also the bladder tissue M2, 3-muscarinic receptor protein levels were examined by immunohistochemistry.

After 12 weeks, the urodynamic test of the test group showed significantly increased non-voiding contractions (10/14 vs 1/13, P<0.05) and decreased compliance ((2.16±0.17) vs (4.18±0.21) ml/cm H₂O, P<0.01) versus the control group.The functional experiments showed that carbachol-induced contractions significantly decreased in hyperlipidemic rabbits than those in controls.As to the relaxant actions of selective muscarinic receptor M3 antagonist solifenacin, the bladder strips of hyperlipidemic rabbits were significantly more sensitive than those of the controls.Histological examination showed thickened intima of internal iliac arteries, urothelial disruption and mild fibrosis in the hyperlipidemic rabbits.Transmission electron microscopy of hyperlipidemic bladder muscle tissues showed swollen mitochondrial damage.As to the immunohistochemical results, compared with the control group, the hyperlipidemic group showed significantly increased M3 receptor expression both in bladder urothelium (146±14 vs 108±9,P<0.01) and smooth muscle layer (131±17 vs 116±15,P<0.05).And the expression was more obvious in urothelium layer.

Involuntary detrusor overactivity with decreased bladder compliance and decreased contractions of detrusor are present in hyperlipidemic rabbits.And chronic bladder ischemia and increased M3 receptor expression may contribute to bladder dysfunction.

To explore the existence of bladder smooth muscle autophagy and its function in cyclophosphamide (CYP)-induced cystitis.

A total of 30 Sprague-Dawley rats were randomized into control (n=6) and experimental groups (4 h, 48 h, 72 h, 96 h,n=24).The experimental group received a single intraperitoneal injection of 150 mg/kg CYPin 0.6 ml saline while the control group had the same volume of saline injection.The changes of microtubule-associated protein light chain 3 (LC3), interleukin-6 (IL-6), IL-1β and tumor necrosis factor-alpha (TNF-α) proteins in bladder muscular layer were measured with Western blot.Co-location of LC3 and alphasmall muscle action (α-SMA) was detected with double-labeled immunofluorescence.Autophagic vacuoles were observed with electron microscopy.The changes in bladder inflammation were measured by hematoxylin and eosin staining.

Autophagy was confirmed in detrusor myocytes with electron microscope, LC3 Western blot and double-labeled immunofluorescence.As compared with the control group, the expression level of LC3 in experimental groups decreased (0.08±0.01 vs 0.13±0.01,P<0.05) at 4 h and increased 1.19, 0.75 and 0.20 fold (0.29±0.04 vs 0.13±0.01,P<0.05;0.23±0.03 vs 0.13±0.01,P<0.05;0.16±0.03 vs 0.13±0.01,P>0.05) at 48, 72 and 96 h respectively.Electron microscopy and double-labeled immunofluorescent also supported the above change of LC3 by Western blot.In addition, as compared with the control groups, IL-6, IL-1β and TNF-α and bladder histological scores of bladder muscular layer significantly increased in experimental groups (all P<0.05).

Autophagy exists in bladder smooth muscle.In cyclophosphamideinduced cystitis, bladder smooth muscle autophagy activation may not be sufficient to protect bladder muscle layer inflammation in rats.

To explore the roles of different casein kinase 2 (CK2) catalytic subunits in androgen receptor (AR) signaling in prostate cancer cell lines.

Prostate cancer cell lines were maintained.Immunofluorescent staining was performed to determine AR nuclear translocation in PC-3/ AR cells with R1881 pretreatment and luciferase gene reporter assay used to determine AR transactivation in LNCaP cells.And reverse transcription-polymerase chain reaction (RT-PCR) was employed to evaluate the mRNA level of prostate-specific antigen (PSA).

R1881-induced AR nuclear localization was reduced significantly (P<0.01).R1881-stimulated ARE-LUC reporter activity in LNCaP cells decreased with reduced level of PSA mRNA, an AR endogenous target (P<0.05).To confirm the target specificity, the gene-specific siRNAs were used for both CK2αand CK2α′subunits.The results of ARE-LUC assay (38.5 vs 31.4) suggested that both CK2 catalytic subunits were involved in androgen-stimulated AR nuclear translocation and AR-mediated gene expression in prostate cancer cells.

Both CK2 catalytic subunits are involved in androgen receptor signaling of prostate cancer cells.

To explore the transfecting effects of RelB small interfering RNA (RelBsiRNA) on murine prostate cancer cell line RM-1 sensitivity of radiotherapy.

The RM-1 cells were divided into RelB-siRNA vector, empty vector, non-transfection and normal groups.After transfection, the first three groups were irradiated by X ray.Clone formation array method was used to measure the surviving fraction and the relevant parameter values after 0, 2, 4, 6, 8 Gy irradiation.Apoptotic rate was measured via Annexin V/PI flow cytometry after 6 Gy irradiation.The level of RelBm RNA was estimated by real-time polymerase chain reaction (PCR) after 6 Gy irradiation.And the RelB protein of cytoplasm and nucleus was detected by Western blot after 6 Gy irradiation.

Clone formation array showed that RelB group at each dose point corresponding to the survival fraction were lower that the empty vector nd non-transfection groups.The D₀, D_qand SF₂values of Rel B group were 1.68, 0.60, 0.43, lower than those of empty vector group 1.92, 3.08, 0.89 and non-transfection group 1.93, 2.76, 0.84.Annexin V/PI flow cytometry demonstrated that the apoptotic rate of RelB group was 15.27%±1.62% and it was significantly higher than the nontransfection group 7.90%±1.50% and the empty vector group 8.40%±0.69% respectively (P<0.01).Real-time PCR indicated that RelB-mRNA amplification of RelB group was 1.18±0.03 and it was significantly lower than the non-transfection group 2.10±0.61 and the empty vector group 1.97±0.66 respectively (P<0.01).Western blot suggested that cytoplasmic gray-scale ratio of RelB group was 0.50±0.08 and it was significantly lower than the non-transfection 1.77±0.19 and the empty group 1.52±0.12 respectively (P<0.01).And the nuclear gray-scale radio of the RelB group was 0.18±0.03 and it was significantly lower than the non-transfection group 0.61±0.12 and the empty group 0.54±0.13 respectively (P<0.01).The Rel B protein inhibition rates of cytoplasmic and nuclear RelB-siRNA were 71.8% and 70.4% respectively.

RelB-siRNA can effectively inhibit the RelB expression of murine prostate cancer cell line RM-1 and significantly increase its sensitivity to radiotherapy.

To explore the prognostic factors of prostate cancer (PCa) patients and evaluate the effect of brachytherapy on survival time.

A total of 289 PCa were recruited to collect their clinical and survival data.And their possible prognostic factors were analyzed.A further comparison of 5-year cumulative survival rate was made between the patients treated by maximal androgen blockade (MAB) and those on MAB plus brachytherapy.

Their median survival time was 73 (7-144) months.And the 1, 3 and 5-year survival rates were 93.1%, 81.0% and 60.2% respectively.Univariate analysis indicated that prostate volume, basal level of prostate-specific antigen (PSA), Gleason score, tumor stage, PSA nadir, time PSA decreasing to nadir and brachytherapy were all predictive factors for survival time.And multivariate analysis further demonstrated that Gleason score, tumor stage and PSA nadir were independent prognostic indicators.And the combination therapy based on brachytherapy could significantly increase the 5-year cumulative survival rate than MAB-based monotherapy.

Gleason score, tumor stage and PSA nadir may predict the prognosis of PCa patients.And brachytherapy significantly improves patient survival.

To determine whether postaurical subcutaneous injection of triamcinolone acetonide is effective for subjective tinnitus refractory to medical treatment.

A total of 100 adult patients with subjective tinnitus of cochlear origin were randomly assigned to receive postaurical subcutaneous.Injection of either triamcinolone acetonide (n=50) or saline solution (n=50).The treatment protocol comprised 5 injections,one per week for 5 weeks.Improvement was measured by tinnitus severity evaluation index evaluation scale,at baseline and one week after the last injection.The follow-up period was 6 months.

No significant difference existed between two groups regarding age,gender,pure tone average,pretreatment tinnitus intensity,tinnitus duration or hearing loss level tinnitus duration (P=0.316,0.685,0.839,0.682 and 0.881 respectively).No significant post-treatment changes in the tinnitus severity index (experimental group=52%,control group=44%) were observed in either group (P=0.724).The most frequently encountered side effect was pain during injection.

The postaurical subcutaneous injection of triamcinolone acetonide has no obvious benefit compared with placebo for subjective tinnitus of cochlear origin refractory to medical treatment.

To characterize the plasma renin activity (PRA) and plasma aldosterone concentration (PAC) and aldosterone/renin ratio (ARR) in patients with aldosterone-producing adenoma (APA).

We retrospectively analysed the data of PRA, PAC and ARR from 80 patients with APA, 70 patients with essential hypertension (EH) and 26 individuals with normal blood pressure (NBP).Patients with hypertension were further divided into taking anti-hypertensive drug group (D) and non drug treatment group (ND).All participants received at least one following tests:ARR screening test, supine-upright position test and saline load test.Receiver-operating characteristic (ROC) analysis was used for exploring the best cut-off value of ARR and low PRA.

The median and percentages of PRA (ng·ml^-1·h^-1,1 ng·ml^-1·h^-1=1μg·L^-1·h^-1), PAC (ng/dl, 1 ng/dl=27.7 pmol/L) and ARR (ng·dl^-1/ng·ml^-1·h^-1) between NBP and EH (ND) groups showed no differences.Over 90% supine PRA≥0.52 and 100% upright PRA≥0.52 in the above two groups.On the contrary, 90% APA (ND) patients upright PRA<0.52.The lowest supine and upright ARR in APA (ND) patients was≥24.2 and≥37.5 respectively.ROC analysis suggested that the best screening cut-off values for APA were supine ARR≥26.0, upright ARR≥37.0; and low PRA cut-off value for APA were supine PRA<0.50 and upright PRA<0.63 respectively.The sensitivity and specificity for APA diagnosis were about 88.2% and 61.5% when PAC≥10.0 after saline load test.

The distinguishing features of PRA, PAC and ARR can be used as a diagnostic indexs for the exclusive diagnosis of APA in various clinical tests.And low PRA cut-off values exist in APA patients.

To explore the expressions of Jumonji domain containing protein 2C (JMJD2C) and hypoxia-inducible factor-1 alpha (HIF-1α) in gastriccarcinoma and their relationship with clinicopathological characteristics.

A retrospective cohort study was performed for 110 gastric cancer (GC) patients at Zhejiang Provincial People′s Hospital from 2005 to 2007.There were 78 males and 32 females with an average age of 57 (32- 79) years.There was no preoperative radiochemotherapy.Immunohistochemical analysis was used to evaluate the expressions of JMJD2C and HIF-1αin 110 specimens of gastric cancer tissues and 80 normal adjacent tissues.

The positive expression rates of JMJD2C and HIF-1αin GC (69.1% (76/110) and 73.6% (81/110)) were significantly higher than those in normal tissues (both 0,both P<0.05).The positive expression of JMJD2C in GC was significantly correlated with TNM stage, invasive depth, lymph node metastasis and distant metastasis (all P<0.05).The positive expression of HIF-1αwas significantly correlated with TNM stage, invasive depth, lymph node metastasis and distant metastasis (all P<0.05).JMJD2C expression was positively correlated with HIF-1αexpression (r=0.219, P<0.05).The survival time of JMJD2C positive group and HIF-1αpositive group were significantly shorter than those of the negative group ((38±4) vs (56±6) months, (38±4) vs (60±6) months, χ²=8.006,7.218, both P<0.01).The survival time of group positive in both JMJD2C and HIF-1αwas significantly shorter than that of single positive or double negative groups (χ²=10.425, P<0.01).

The over-expressions of JMJD2C and HIF-1αin gastric cancer tissues play a role in the growth, invasion and metastasis of gastric tumor.Both may be used to predict the prognosis of GC patients.

To explore the expressions of S100A8 and S100A9 in gastric cancer.

A total of 176 patients with gastric cancer, including 124 males and 52 females, were recruit from 1998 to 2004,average age 57 (26-80) years.The expressions of S100A8 (n=125), S100 A9 (n=176) and S100A8/A9 heterodimer (calprotectin) in gastric tissue samples were assessed by immunohistochemistry and immunofluorescence.The co-localization of S100A9 and its dimerization partner S100A8 and heterodimer S100A8/A9 were examined by laser confocal scanning.Receiver operator characteristic (ROC) curve was used in determining the cut-off value of S100A9 and S100A8-positive inflammatory cell counts in evaluating the pathological TNM stage.To obtain associations between S100A9 or S100A8 cell counts and clinicopathologic variables, the data were cross-tabulated and χ²-test was performed.Cumulative survival was estimated by the Kaplan-Meier method.

ROC curves using the S100A9 and S100A8-positive inflammatory cell counts were 0.623 and 0.522 for pathological TNM stages respectively.The cutoff values were 200 and 65 per 200× magnification field with a sensitivity of 61.48% and 51.09% and a specificity of 64.29% and 51.52% respectively.Patients with S100A9 positive expression (n=77) had better overall survival than negative expression (n=99)((35.1±10.8) vs (20.3±3.0) months, P=0.021).There was no statistical significance between S100A8 positive expression (n=62) and negative expression (n=63)((26.4±2.8) vs (29.5±2.9) months, P=0.145).In gastric cancer tissues, both S100A9 and S100A8 proteins were detected in tumor-infiltrating inflammatory cells while no case of S100A8/A9 heterodimer was found.In addition, S100A9 and S100A8 proteins were detected in inflammatory cells in chronic gastritis.Distribution of these two proteins also partly overlapped.

S100A9 positive expression in gastric cancer tissues is associated with an excellent prognosis.But S100A8 positive expression has no prognostic association.Calprotectin expression differs between gastric cancer and gastritis.Further explorations are warranted.

To explore the clinicogenetic characteristics and protein status of parathyroid carcinoma (PC) diagnosed at Peking Union Medical College Hospital from January 1980 to December 2012.

Genomic DNA was extracted from peripheral blood lymphocytes and paraffin-embedded tissue for analysis.The mutations of CDC73 gene were detected by direct sequencing.And the expression of parafibromin in tumor tissues was evaluated by immunohistochemical analysis.

Twenty-four PC patients were recruited.The mean ages at the diagnosis of PHPT and PC were 42.2 and 42.6 years respectively.The serum calcium at the diagnosis of PHPT was 3.78 mmol/L, serum phosphorus 0.65 mmol/L and the median increment of serum parathyroid hormone (PTH) level 20.0.The recurrent rate was 76.2% during a 5-year followup. Genetic analysis identified 11 mutations of CDC73 gene (45.8%),among which c.34_35 insCT, c.626_629 del AACA,c.260_261 delGA,c.570 delG,c.40 C>T and IVS3+1G>A were novel mutation first identified in our cohort.Nine of them had germline mutations.All tissue samples from normal parathyroid displayed strong positive immunostaining of parafibromin.Complete (55%,11/20) or partial (45%,9/20) loss of parafibromin expression was observed in PC tissues.The age at the diagnosis of PHPT,serum calcium,serum phosphorus,PTH,alkaline phosphatase (ALP),bone involvement rate,kidney stone/calcification rate,recurrent rate,metastatic rate and immunostaining level showed no significant difference between the patients with and without CDC73 mutation.

In Chinese population, CDC73 mutation and complete/partial loss of immunohistochemical staining for parafibromin occur frequently in PC.Therefore it may be useful in the subset of tumors with equivocal histological examination.

To explore the effects of low-dose insulin on oxidation-reduction of heart and kidney in rats immediately after severe burns.

Twenty four male Sprague-Dawley (SD) rats were randomly divided into 3 groups of sham burn, burn and treatment (n=8 each).The sham burn group was placed into 37℃warm water for 15 seconds to simulate burn process and received no fluid replacement.The burn and treatment groups were immmersed into (95±0.5)℃ hot water for 15 seconds to make a rat model of 30% total burn surface area,Ⅲdegree burn injury and immediately received an intraperitoneal injection of physiological saline (40 ml/kg).At the same time, a subcutaneous injection of insulin (1.0 U·kg^-1·d^-1) was given in the treatment group and a subcutaneous injection of same-volume physiological saline in the burn group.The rats were sacrificed after 24 post-scald hours (PSH).Abdominal aortic blood was collected for an analysis of blood glucose.The oxidation and antioxidation parameters of heart and kidney, such as malondialdehyde (MDA), xanthine oxidase (XO), myeloperoxidase (MPO), total superoxide dismutase (T-SOD), superoxide dismutase 1, 2 (SOD1, 2), catalase (CAT) and glutathion peroxidase (GPx), were detected by spectrophotometry.

Compared with the sham burn group, MDA content, XO and MPO activities of heart and kidney were significantly higher in the burn group (all P<0.05); in the treatment group, MDA content and XO activity of heart were significantly lower than the burn group ((0.85±0.07) vs (1.11±0.07) nmol/mg, (69.72±1.94) vs (77.21±2.10) U/g) while the MPO activities of heart and kidney were significantly lower (all P<0.05).compared with the sham burn group, the activities of T-SOD, CAT, GPx of heart and kidney were significantly lower in the burn group, SOD1 activity of kidney was significantly lower, but SOD2 activity of kidney was significantly higher while SOD2 activity of heart was significantly lower (all P<0.05); Compared with the burn group, the activities of T-SOD and SOD1 of heart in the treatment group were significantly higher ((83.5±2.5) vs (79.6±3.2), (62.8±2.3) vs (58.8±3.0) U/mg),CAT and GPx activity of heart and kidney were significantly higher (all P<0.05).Compared with the sham burn group, blood glucose in the burn and treatment group were significantly higher ((7.81±0.30), (7.19±0.22) vs (6.30±0.24) mmol/L) and blood glucose in the treatment group was significantly lower than the burn group ((7.81±0.30) mmol/L)(all P<0.05).

During an early stage, a low-dose insulin may intervene in heart tissue lipid peroxidation of severely burned rats.And differences exist in the effects of oxidation-reduction between heart and kidney.

To explore the effects of nicotinic acid intervention on vascular endothelial dysfunction mediated by oxidized low density lipoprotein (ox-LDL) /lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) in diet-induced obese immature rats and its possible mechanism through detecting the expression levels of ox-LDL and LOX-1 in abdominal arota.

A model of diet-induced obese immature rats was established by high-fat diet.And 30 immature rats were divided randomly and equally into control (n=10), high-fat (n=10) and drug control (n=10) groups.At the end of 12 weeks, the levels of serum total cholesterol (TC), triglyceride (TG), LDL and high-density lipoprotein (HDL) were examined.The levels of ox-LDL, soluble intercellular adhesion molecule-1 (sICAM-1), endothelin and nitric oxide (NO) were detected.The gene and protein expressions of LOX-1 and ICAM-1 in abdominal aorta were detected.And the location protein expressions of LOX-1and ICAM-1 were examined.

High-fat diet induced hyperlipidemia and obesity in immature rats.The serum levels of TG, TC, LDL, ox-LDL and endothelin in high-fat and drug control groups were all higher than control group ((0.98±0.12) and (0.69±0.06) vs (0.49±0.06) mmol/L, (2.11±0.16) and (1.62±0.12) vs (1.30±0.12) mmol/L, (0.71±0.04) and (0.50±0.03) vs (0.30±0.04) mmol/L, (44.2±5.1) and (33.7±2.1) vs (26.6±2.9) μg/L, (187±10) and (157±6) vs (118±7) pg/ml).The indices in high-fat group were higher than those in drug control group (all P<0.01).The levels of HDL and NO in high-fat and drug control groups were lower than those in control group (all P<0.01); the levels of HDL and NO in high-fat group lower than those in drug control group (all P<0.01).And the levels of LOX-1, ICAM-1 protein and mRNA in high-fat group were higher than those in drug control and control groups (all P<0.01).ox-LDL was correlated positively with LOX-1, TC, TG, LDL, endothelin and ICAM-1 (r=0.918, 0.867, 0.857, 0.834, 0.869, 0.644, all P<0.01), but negatively with NO and HDL (r=-0.823,-0.872, P<0.01).

Early treatment of nicotinic acid can protect endothelial function through inducing therapeutic effects on hyperlipidemia and antioxidation and down-regulating the expression level of ox-LDL/LOX-1 in vascular endothelium.